, BOSTON PUBUC LIBRARY I GOVBRNMENT DOCUWENTS UEPARTWESST RECEIVED ] FEB 1 6 2000 I The Pesticides Monitoring Journal is published quarterly under the auspices of the FEDERAL WORKING GROUP ON PEST MANAGEMENT (responsible to the Council on Environ- mental Quality) and its MONITORING PANEL as a source of information on pesticide levels relative to man and his environment. The WORKING GROUP is comprised of representatives of the U.S. Departments of Agricul- ture; Commerce: Defense; the Interior; Health, Education, and Welfare; State; Transportation; and Labor; and the U.S. Environmental Protection Agency. The pesticide MONITORING PANEL consists of representatives of the Agricultural Research Service, Animal and Plant Health Inspection Service, Extension Service, Forest Service. Depart- ment of Defense, Fish and Wildlife Service. Geological Survey, Food and Drug Administration, Environmental Protection Agency, National Marine Fisheries Service, National Science Founda- tion, and Tennessee Valley Authority. Publication of the Pesticides Monilorini> Journal is carried out by the Technical Services Divi- sion, Office of Pesticides Programs of the Environmental Protection Agency. Pesticide monitoring activities of the Federal Government, particularly in those agencies repre- sented on the pesticide MONITORING PANEL which participate in operation of the national pesticides monitoring network, are expected to be the principal sources of data and interpretive articles. However, pertinent data in summarized form, together with interpretive discussions, are invited from both Federal and non-Federal sources, including those associated with State and community monitoring programs, universities, hospitals, and nongovernmental research institu- tions, both domestic and foreign. Results of studies in which monitoring data play a major or minor role or serve as support for research investigation also are welcome; however, the Journal is not intended as a primary medium for the publication of basic research Manuscripts received for publication are reviewed by an Editorial Advisory Board established by the MONITORING PANEL. Authors are given the benefit of review comments prior to publication. Editorial Advisory Board members are: John R. Wessel, Food and Drug Administration, Chairman Paul F. Sand, Agricultural Research Service, Vice Chairman Anne R. Yobs, Center for Disease Control William F. Durham, Environmental Protection Agency Thomas W. Duke, Environmental Protection Agency G. Bruce Wiersma, Environmental Proteclion Agency William H. Stickel, Fish and Wildlife Service Milton S. Schechter, Agricultural Research Service Herman R. Feltz, Geological Survey Mention of trade names or commercial sources in the Pesticides Monitoring Journal is for identification only and does not represent endorsement by any Federal agency. Address correspondence to: Paul Fuschini Editorial Manager PESTICIDES MONITORING JOURNAL U.S. Environmental Protection Agency Room B49 East, Waterside Mall 401 M Street, S.W. Washington, D. C. 20460 Martha Finan Joanne Sanders Editors CONTENTS Volume 8 June 1974 Number 1 PESTICIDES IN PEOPLE Epidemiology of orgatioclilorinc insecticides in the adipose tissue of Israelis M. Wassermarm, L. Tomatis. Dora Wassermann, N. E. Day, Y. Groner, S. Lazarovici. and Deborah Rosenfeld RESIDUES IN FOOD AND FEED Polyclilorinated biplienyl and organoclilorine pesticide residues in Canadian chicken eggs Page Jos Mes, D. E. Coffin, and D. Campbell RESIDUES IN FISH, WILDLIFE, AND ESTUARIES DDT plus PCB's in bluhher of liarhor seals 12 Raymond E. Anas Chlorinated hydrocarbon and mercury residues in woodcock in the United Slates. 1970-71 15 Donald R. Clark, Jr., and M. Anne Ross McLane Studies on the distribution and flux of pesticides in waterways associated with a ricefield — marshland ecosystem 23 Thomas M. Ginn and Frank M. Fisher, Jr. Selected chlorinated hydrocarbons in bottom material from streams tributary to San Francisco Bay 33 LeRoy M. Law and Donald F. Goerlitz Organochtorine residues in golden eagles. United Stales — March 1964-Jidy 1971 37 Russell F. Reidinger, Jr., and D. Glen Crabtree Toxaphene content of estuarine fauna and flora before, during, and after dredging toxaphene-contatninated sediments 44 Robert J. Reiniold and Charles J. Diirant GENERAL Restnethrin residues in foliage after aerial application 50 Theresa L. Andrews Detection of DCPA residues in environmental .samples 53 F. M. Miller and E. D. Gomes Degradation of four organophosphate insecticides in grape tissues 59 Wray Winterlin, Charles Mourer, and J. Blair Bailey APPENDIX Chemical natnes of compounds discussed in this issue 66 ERRATA 67 PESTICIDES IN PEOPLE Epidemiology of Organochlorine Insecticides in the Adipose Tissue of Israelis ' M. Wassermannr L. Tomatis.' Dora Wassermann,- N. E. Day,^ Y. Groner,= S. Lazarovicir' Deborah Rosenfeld = ABSTRACT This paper leporls the findings obtained in 1967-71 during an investigation of organochlorine insecticides (OCJ) storage in the adipose tissue of Israelis. Specimens of adipose tissue (307) collected during imtopsy from Israelis who had no known occupational exposure to organochlorine insecticides were analyzed by the gas chro- matographic method for organochlorine insecticides (DDT- derived material: alpha, beta, and gamma isomers of BHC, dieldrin, and heptachlor epo.xide). Findings indicate a positive age association for DDT-derived material stored in the adipose tissue of Israelis of both .w.xes. Males generally were found to store higher amounts of •y.p'-DDT and total DDT than females. Comparison of adipose tissue from stillborns with tissue 'ram infants showed that DDT increased in the first months )/ postnatal life, but storage levels of BHC. dieldrin. and heptachlor epoxide remained approximately the .same. DDT 'evels continued to rise with age levels, except for a slight lecrea.se in the 24-through-44-year-olds. The highe.st levels )/ DDT were found in the age group of 70 and over: sec- "id highest were among 45-to-69-year-olds. These findings It Israel differ from the authors' earlier findings in South Africa, Thailand. Nigeria, and Brazil, which revealed the lighcft concentrations of OCI in the 24-lhrough-44-vear- This research study was supported by the U.S. Department of Health, liducation and Welfare. Public Health Service, National Communi- cable Disease Center, Atlanta, Ga., Research Grant No. BSS-CDC- IS-9, and by an agreement of the World Health Organization— Inter- national Agency for Research on Cancer. Department of Occupational Health, Hebrew University— Hadassah Medical School, Jerusalem, Israel. World Health Organization- Cancer, Lyon, France. International Agency for Research on /OL. 8, No. 1, June 1974 A positive age association with DDT storage in all ages was observed in 1965 aiul 1967 surveys by the authors in people from Kenya and Israel, and by Davies and Milby in the nonwhite population of the USA. In the countries studied, the storage level of DDT and de- rived material increases with age in the general population up to the age of 45, and either rises or falls after 45 years, depending on the country. This leads the authors to the opinion tliat the age group of 25 through 44 years may be the most suitable indicator of DDT storage levels in a community. A positive relationship between p,p'-DDT and dieldrin stor- age was also noted. Introduction The presence of DDT and its metabolite DDE in the adipose tissue of the population of Israel was first dem- onstrated in a study carried out by Wassermann et al, (/) in 1963-64, They revealed that these chemicals were stored in the adipose tissue of people who had no known occupational exposure to the chemicals and suggested that the DDT storage process is influenced by individual factors such as sex and age. A further study carried out by the same investigators in 1965-66 (2) ascertained that Israelis over 10 years of age stored higher amounts of DDT-derived material than did those under 10 years of age. Males over 10 stored higher amounts of DDT-derived materials than did females. A study by Polishuk, Wassermann, et al, (3) on pregnant Israeli women indicated less storage of DDT-derived 1 material, BHC isomers, and dieldrin than m nonpreg- nant women of the same age. The organochlorine in- secticides (OCI) found in the fat tissue of pregnant women were present also in the maternal and fetal blood in all cases studied. These findings suggested that, in pregnancy, the metabolism of organochlorine insec- ticides is enhanced and that they pass the placental barrier. Data published up to the mid-sixties on the storage of DDT-derived material in the adipose tissue of people from various countries affirmed the concept of organo- chlorine insecticides as "current constituents of the human body" (4). Other studies revealed the impact exerted on OCI storage in man by individual factors such as age, sex, and race (1. 2. 5-7/), by particular physiological states (3. 12). as well as by living and working conditions (7, 13). Concern for the potential hazard to human health by organochlorine insecticide accumulation in the body in- creased following reports that OCI are potent hepatic microsomal inducers which may quantitatively alter the response to various drugs and toxic compounds as well as to naturally occurring substances in the animal body. This may lead to alteration of homeostasis of biochem- ical (endocrine, immunologic, etc.) processes (i. 14-16). Experimental evidence of the capacity of OCI to increase tumor incidence in laboratory animals was also reported (17-19). Thus it became necessary to assess the size and trends of OCI storage in populations in general and to find which age group would be most useful for studying this storage phenomenon in various areas of the world. This paper reports the findings of a study of OCI storage in Israelis and compares the data to those obtained by the same investigators in various populations in Africa. Asia, and South America in the framework of a program launched by the World Health Organization Interna- tional Agency for Research on Cancer, Lyon, France. Methods and Materials A total of 307 samples of adipose tissue were collected from 1967 to 1969 during autopsy from the subcuta- neous fat of the abdominal wall of persons who had no known occupational exposure to pesticides. The distri- bution of samples according to age and sex is shown in Table 1. Samples of 1-2 g of adipose tissue were col- lected in jars containing 10% formalin. Specimens ot 500-mg adipose tissue were extracted three times with a total of 20 ml of petrol ether and cleaned by means of a Kontes Co-Distiller. The extract was reduced to 0.5 cc, from which 5-20 /xl were injected into a Micro- tek MT-220 gas chromatograph equipped with dual electron-capture detector and strip chart recorder. A 6-ft U-shaped glass column packed with 5% QF-1 60-80 mesh chromoport xxx and a 4-ft U-shaped g\i column packed with 10% SE-30 on 60-80 mesh chi moport xxx were used. A mixture of pure organochlori insecticides was used as standard; the concentration each compound was 0.1 ppm. Recovery was about 85 for the compounds identified in this study. The sensitiv j of detection was 0.1 to 0.3 )u,g/kg wet weight for thej compounds. TABLE 1. -Disliibution of human adipose tissue samples age and se.x of subject AOE Males Females Total Stillborns 0-11 months 5-24 years 25-44 years 45-69 years 70 and over Total 26 22 38 29 32 27 174 18 18 23 24 31 19 133 44 40 61 53 63 46 307 Results and Discussion In 307 samples of adipose tissue collected from Isra^i who had no known occupational exposure to organ chlorine insecticides, gas-chromatographic analysis vealed the presence of DDT-derived material; alp beta, and gamma isomers of BHC; dieldrin; and hefi chlor epoxide. It must be stressed that, as noted in previous stuc. (20-23), marked individual variations in storage le\ were observed among individuals of the same sex age group who were not occupationally exposed OCI's. Similar individual variations were observed experimental animals kept in experimental condition; which all were exposed to the same level of DDT {1 The broad spectrum of age groups and the relati\ large number of cases investigated in this study em us to follow up the dynamics of OCI storage in population by age and sex. Concentration of OCI in the adipose tissue of the s born group averaged: total DDT 0.7 ppm; total B 0.04 ppm; dieldrin 0.02 ppm; and traces of heptacl epoxide below O.OI ppm, thus indicating the accum tion of these compounds during fetal life (Tables 2-4 The first months of life in the external environment to an increased DDT storage. In the age group ur 11 months, total DDT averaged 5.8 ppm; the stoi level of BHC, dieldrin, and heptachlor epoxide remai about the same as in stillborn. DDE averaged 51. of total DDT-derived material. The mean total p.p'-E was 5.7 ppm and the mean total a.p'-DDT was ( ppm. There was a significant difference in the stoi of p.p'-DDT and total DDT between the stillborn the 0-through- 11 -month age group (p < 0.01). Pesticides Monitoring Jour O 00 vt — O 00 -H (-1 00 rJ 8g OO 00 O Tf 82 SI iS vo — 00 -ft 00 r- O Tt NO r- 00 Tj o — 32 o ri O r- r^ p-> (N OO 0\ T^ ri o Tt 00 o rJ -. 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Total BHC was 0.5 ppm, dieldrin was 0.14 ppm, and heptachlor epoxide was 0.01 ppm. From these data it was concluded that the storage level of DDT and derived material increased with age, the highest values being found in the 70-year-and-over age group. Tables 3 and 4 also showed that males stored higher amounts of p,p'-DDT and total DDT in all the age groups, except the 0-through-l 1 -month age group in which the DDT storage levels were of about the same order: 5.34 ppm total DDT in males and 5.82 ppm total DDT in females. The authors found the trend for a positive age associa- tion of DDT storage in populations living in various areas of the world. In the general populations of South Africa {25). Thailand (23). Nigeria (22). and Brazil (20), the 25-through-44-year age group stored the highest amount of OCI in both sexes, after which there was a decrease in the storage level. In the general population of Uganda, the storage level of DDT was comparable in the 5-through-24- and 25-through-44-year age groups (26). In the general populations of Kenya (21), Israel (2. and this paper), and nonwhite populations of the USA (8), it appears that a positive age association with DDT storage occurs in all ages. These data suggest that the level of the OCI storage process tends to vary after the age of 45 years; in some populations it is higher and in others it is lower than i the 25-through-44-year age group. It appears reasonabl therefore, to consider the 25-through-44-year age groi the best indicator of OCI storage in a community f( purposes of comparison (Table 5). TABLE 5. — Storage of DDT-derived material in adipo tissue of 25-tlirougli-44-years-olds in several countries Total DDT, PPM Country References M .itF Males Females Uganda 2.9 35 1.2 Wassermann el al. (2 Kenya 4.5 4.6 4.2 Wassermann et al. (2 Nigeria 6.5 7.4 5.9 Wassermann et al. (2 Brazil 7.8 9.6 6.3 Wassermann et al (z South Africa Wassermann el al (I White 8.5 10.5 6.6 Bantu 6.5 8.6 4.4 Thailand 13.0 15.5 10.2 Wassermann et al C Israel 14.4 15.9 12.2 This paper. In the study carried out on OCI storage in the adipc tissue of people from Nigeria (.22), a positive age ass elation up to the age of 45 years was found both f DDT and dieldrin. These findings confirmed expe mental data the authors had obtained in rats submitt to a high dosage of p,p'-DDT: namely, a parallel crease in the storage of dieldrin, although these anim: had not received additional dieldrin apart from tt naturally in food and water (27). In the present stuc authors compared the concentration of dieldrin in t adipose tissue of people having; (1) high concentration p,p'-DDT, and (2) low concentration of p.p'-DDT. The statistical analysis revealed increased storage dieldrin (p < 0.01) in the group with a higher conci tration of p.p'-DDT. These findings may be explained by a biochemical int relationship of the two compounds in the animal boi the presence of a large amount of DDT interfering w the detoxication of dieldrin resulting in its accumulat in adipose tissue. LITERATURE CITED 1 1) Wassermann. M.. M. Gon. D. Wassermann. and L. / lermayer. 1965. DDT and DDE in the body fat people in Israel. Arch. Environ. Health 11:9, 375-3 (2) Wassermann. A/.. D. Wassermann. L. Zellermayer, . M. Gon. 1967. Storage of DDT in the people of Isr Pestic. Monit. J. 1:2. 15-20. (3j Polisluili. Z. W.. M. Wassermann. D. Wassermann, Groner, S. Lazarovici, and L. Tomatis. 1970. Eff of pregnancy on storage of organochlorine insectici' Arch. Environ. Health 20:2, 215-217. (4) Wassermann. M. and D. Wassermann. 1966. Reco tion of a new group of toxic substances as curi constituents in the human body: the organochlo insecticides. Proc. XV Int. Cong. Occup. Health, Vier Austria, 6:2, 954. (5) Brown J. R. 1967. Organochlorine pesticide resii in human depot fat. Can. Med. Ass. J. 97:367-372. Pesticides Monitoring Jour '6) Davics, J. E.. W. F. Edmundsuit. N. J. Schneider, and J. C. Cassady. 1968. Problems of prevalence of pesti- cide residues in humans. Pestic. Monit. J. 2:80-85. 7) Davies, J. E., and J. H. Milby. 1969. Epidemiology of pesticides. In: Mrak, E. M. Report of the secretary's commission on pesticides and their relationship to en- vironmental health, Parts I and II. U.S. Department of Health, Education and Welfare. '8) Fiseidva-Bcrgennu, V .. J. L. Kadom.\ki, J. E. Davies, and J. H. Da\i.s: 1967. Levels of chlorinated hydrocar- bon pesticides in human tissue. Ind. Med. Surg. 36: 65-70. 9) Hunter. C. G.. J. Robinson, and A. Richardson. 1963. Chlorinated insecticide content of human body fat in southern England. Brit. J. Med. 1:221-224. 0) Robinson. J.. A. Ricliardson, C. G. Hunter , A. N. Crab- tree, and H. J. Rces. 1965. Organochlorine insecticide content of human adipose tissue. Brit. J. Ind. Med. 22:220-229. 1) Zavon, M. R., R. Tyre, and L. Laforre. 1969. Chlo- rinated hydrocarbon insecticide content of the neonate. Ann. N.Y. Acad. Sci. 160:196-200. 2) Curley. A. and R. Kimbrouglt. 1969. Chlorinated hydro- carbon insecticides in plasma and milk of pregnant and lactating women. Arch. Environ. Health 18:156-164. ?J Hayes. W . J.. Jr., G. E. Quinby. K. C. Walter, J. W. Elliott, and W. M. Upliolt. 1958. Storage of DDT and DDE in people with different degrees of exposure to DDT. Arch. Industr. HIth. 18: 398-406. f) Conney, A H., R. Welch, R. Kuntznuin. R. Chang, M. Jacobson, A. D. Munro-Faure, A. W. Peck, A. B\e, A. Poland, P. J. Poppers, M. Finster, and J. A. Wolff. 1971 . Effects of environmental chemicals on the metab- olism of drugs, carcinogens and normal body constit- uents in man. Ann. N.Y. Acad. Sci. 179:155-172. ')) McLean, A. E. M., and E. K. McLean. 1969. Diet and toxicity. Brit. Med. Bull. 25:278-281. )j Wasscrmann, M.. D. Wassernumn, E. Kedar, and M. Djavaherian. 1971. Immunological and detoxication interaction in p.p'-DDT fed rabbits. Bull. Environ. Contam. Toxicol. 26:426-435. ') Nagasaki, H., S. Toniis, T. Mega, M. Maruganii, and N. ho. 1971 . Development of hepatomas in mice treated with benzene hexachloride. Gann 62:431. (IS) Toinatis, L., V. Turusov, N. Day, and R. T. Charles. 1972. The effect of long-term exposure to DDT on CF-I mice. Int. J. Cancer 10:489-506. (19) Turusov, V. S., N. E. Day, L. Tomatis, E. Gati, and R. T. Charles. 1973. Tumors in CF-I mice exposed for six consecutive generations to DDT. J. Nat. Cancer Inst. 51:983. (20) Wasscrmann, M., D. P. Nogueira, L. Toinatis, N. E. Day, E. Athie, D. Wasscrmann, M. Djavaherian, and C. Guttel. 1972. Storage of organochlorine insecticides in people of Sao Paulo, Brazil. Ind. Med. Surg. 41:3 22. (21) Wasscrmann, M., M. G. Rogofj, L. Tomatis, N. E. Day, D. Wasscrmann, M. Djavaherian, and C. Guttel. 1972. Storage of organochlorine insecticides in the adipose tissue of people of Kenya. Ann. Soc. Beige Med. Trop. 52:6, 509-514. l22) Wasscrmann, M.. G. O. Sojoluwe. L. Tomatis, N. E. Day, D. Wasscrmann, and S. Lazarovici. 1972. Storage of organochlorine insecticides in people of Nigeria. Environ. Physiol. Biochem. 2:59-67. (23) Wasscrmann. M..M. Trishnanatutu, L. Tomatis, N. E. Day, D. Wasscrmann, V. Rungpitarangsi, V. Chiam- sakol, M. Djavaherian, and S. Cucos. 1972. Storage of organochlorine insecticides in the adipose tissue of peo- ple in Thailand. The Southeast Asian I. of Trop. Med. and Public Health 3:2, 280-285. (24) Tomatis, L., V. Turusov, B. Terracini, N. Dav, W. Barthcl, R. T. Charles, G. B. Collins, and M. Boiocchi. 1971. Storage levels of DDT metabolites in mouse tis- sues following long term exposure to technical DDT. Tumori 57:377-396. (25) Wasscrmann, M., D. Wasscrmann, 5. Lazarovici, A. M. Coetzce. and L. Tomatis. 1970. Present state of the storage of the organochlorine insecticides in the general population of South Africa. S. Afr. Med. J. 44:646-648. (26) Wasscrmann, M., L. Tomatis, D. Wasscrmann, N. E. Day, and M. Djavaherian. Storage of OCl in the adi- pose tissue of Ugandans. Bull. Environ. Contam. Toxi- col. (In press). (27) Wasscrmann, M., D. Wa.sscrmann, and S. Lazarovici. 1969. Effects of thyroidectomy on the storage of organo- chlorine insecticides. Bull. Environ. Contam. Toxicol. 4:6, 327-336. :l. 8, No. 1, June 1974 RESIDUES IN FOOD AND FEED Polychlorinated Biphenyl and Organochlorine Pesticide Residues in Canadian Chicken Eggs ^ Jos Mes, D. E. Coffin, and D. Campbell ABSTRACT A nationwide survey in Canada of polychlorinated biphenyl (PCB) and organochlorine pesticide residues in eggs revealed an average of less than 10 pph for both groups of compounds. PCB's and p.p'-DDE were found in all samples: at least 95 percent also contained dieldrin and p.p'-DDT. Lindane and cis- and Irans-chlordane were present in 75 percent of ail eggs. No significani differences were observed among the different regions of the country. hilrodiulion The presence of polychlorinated biphenyls in the envi- ronment in general and the food supply in particular has been a growing concern for several years {1.2). A recent incident of PCB contamination (0.6-1.9 ppm) of thousands of eggs in the United States, due to contami- nated poultry feed, is evidence of this environmental problem (3). A nationwide survey of PCB's in domestic chicken eggs in Canada was therefore undertaken; initial samples were collected in 1971. The analytical data presented in this paper were obtained from the liquid portion of the egg only. This paper pre- sents data on PCB's and several organochlorine pesti- cides. Sampling Procedure Twenty dozen grade A eggs were collected from each of the following five regions of the country: Eastern (New- foundland, Prince Edward Island, Nova Scotia, and New ' Health Protection Branch of the Department of National Hcilth and Welfare, Tunney's Pasture, Ottawa, Canada. Brunswick); Quebec; Ontario; Central (Manitoba an Saskatchewan); and Western (Alberta and British Cc lumbia). The stipulation was that all eggs from a specific regie be produced locally. The eggs were stored at 8°C unt analyzed. Analytical Methods All solvents were of a glass-distilled, residue-free grad and were checked for purity. Pesticides and decachlon biphenyl standards were 99 percent pure as verified t gas chromatography (GO. Aroclor 1260 was used i supplied by the manufacturer, Monsanto Chemical Con pany. EXTRACTION The liquid content of 12 eggs was pooled and stirre with a glass rod until thoroughly mixed. A 50-g samp was extracted for 3 min in a soil dispersion mixer (St Manufacturing Co., St. Louis. Mo.) with a 300-ml mi ture of hexane and acetone (2:1 v/v), previously warmt to 40°C. The extract was filtered through prewasht anhydrous Na2S04 to remove the water, concentrati on an all-glass rotary evaporator (<25"C), and dilut up to 50 ml. A 1-ml aliquot was evaporated in a pi weighed aluminum dish to determine lipid content. CLEANUP AND SEPARATION OF PCB'S AND PESTICIDES An aliquot equivalent to approximately 2 g of lipid w subjected to the low-temperature micro precipitati technique (4) and further cleaned on a 5 percent Fieri Pesticides MoNixoiaNG Journ column (deactivated with 5 percent distilled water after heating overnight at 140°C) (5). PCB's and pesticides A'ere partly separated on a silicic acid column according :o Armour and Burke (6), except that the silicic acid iivas prewashed with the eluent used for the pesticides, neated overnight at 130°C, and deactivated with 5 Dercent distilled water. A final cleanup for the pesti- :ide fraction was carried out as before on a 5 percent i^Iorisil column. IDENTIFIC.MIUN AND QU.'\NTIFICATION The PCB fraction was concentrated to 1 ml; the pesti- ;;ide fraction was carefully evaporated to dryness under ii gentle stream of No and redissolved in 1 ml of hexane. \ 5-jjA aliquot was injected into a Varian Aerograph ieries 1400 gas chromatograph with an electron-capture letector (tritium foil) under the following conditions: Column: ',4-in.-by-6-ft glass, packed with 6 percent OV-210 + 4 percent SE-30 on Chromosorb W(AW) 60/80 (0.6 g OV-210 + 0.4 g SE-30 + 10 g solid support) femperatures: Injector 220°C Column 212°C Detector 225 °C 'o give a retention time of 13 min for p.p'-DDT an pproximate flow rate of 50 ml No/min was used, tandard solutions were made up to contain 50x10 " '.g/fil of Arocior 1260 (5 yiil were equivalent to a /2 (full-scale deflection) (FSD) for the highest peak on 1-mV recorder) or 0.5-5.0x10-" fj.g/ fjL\ of pesticide, epending upon the individual response of the pesti- ide (e.g., 5x10-' fxg/ fi\ of p.p'-DDT). A 10-/^1 aliquot f the standard pesticide solution had a Vi FSD for ,p'-DT. . 5-fi\ injection of standard solution was made before nd after every two sample injections. CB's and pesticides were quantitated by using peak- sights. Peaks 8. 10, and 11-15 in Arocior 1260, accord- ig to the numbering system of Reynolds (7) and the rganization for Economic Cooperation and Develop- lent (O.E.C.D.) (8). were used for quantification. ONFIRMATION OF PCB'S he PCB fractions of 64 samples were pooled to give total of approximately 13 /xg of PCB. hin-layer chromatography (TLC) was carried out on •ecoated aluminum oxide (type E) Fn-^ 20-by-20-cm lates (Brinkman Instruments, Ltd., Canada), activated 110°C for 1 hr. A total of 2.5 /iig of PCB from the loled sample was spotted at a concentration of 0.25 ?/spot; two reference spots of Arocior 1260 at 2.5 j/spot were applied at each end of the line of origin, le TLC plate was developed in 1 percent acetone in OL. 8, No. 1, June 1974 hexane and the reference spots were made visible with AgNO,.j (9). The standard Arocior 1260 showed two spots and the corresponding areas of the sample, as well as a blank, were scraped, eluted with hexane, and gas-chromatographed as above. A portion of the pooled sample of PCB equivalent to 1 fig of PCB was perchlorinated and the resultant deriv- ative was identified by GC according to Berg et al. (10), except that a Griftin-Worden pressure vessel (Kontes Glass Co. k-767100) was used for perchlorination. CX)NFIRMATION OF PESTICIDES All pesticide fractions, besides having been gas-chro- matographed as before, were also run on a dilTerent column under the following conditions: Column: ' 4 -in.-by-6-ft glass, packed with 5 percent QF-I on Chromosorb W(AW) 60/80 (0.5 g QF-1 4- 10 g solid support) Temperatures: Injector 208 °C Column 175°C Detector 229°C To give a retention lime of 26 min for p.p'-DDT, a flow rate of approximately 40 ml N._./min was used. The pesticide fractions of every other 10 samples were pooled and chromatographed on TLC plates as above. The five pooled samples were spotted at a concentration of 1.2-1.6 /xg/spot of estimated pesticide; two reference spots of an appropriate standard pesticide mixture (2.5 yug/spot for an individual pesticide) were applied at each end of the line of origin. After development and visuali- zation of the standards, the TLC plate was divided into five areas corresponding to the following pesticides, in order of increasing Rf value: 1. dieldrin, heptachlor epoxide, and lindane 2. cis- and trans-chlordane and p,p'-TDE 3. p,p'-DDT 4. DDMU, o,p'-DDT. o,p'-DDE, and heptachlor 5. p,p'-DDE, aldrin Each area was then subdivided into six equal portions corresponding to the five pooled samples and a blank. Adsorbent from these portions was removed, and the pesticides were eluted with hexane and rechromato- graphed on both GC columns. CONTROLS Samples were spiked by adding 1 ppm and 1-10 ppb levels of Arocior 1260 and pesticides to 50 g of whole liquid egg before extraction. At different times during the survey three blanks were run through the complete analytical procedure, starting with a simulated extraction using the same solvent mix- ture used for the egg samples. Results and Discussion Results in Table 1 indicate that the mean level of all residues was below 10 ppb. A PCB and pesticide prob- lem in eggs on the Canadian market apparently does not exist. TABLE 1. — PCB and pesticide residues in whole liquid Canadian chicken eggs Compound Average PPB > Maximum PPB OBSERVED Percentage OF SAMPLES CONTAINING RESIDUES PCB's as Aroclor 1260 Lindane Heptachlor Heptachlor epoxide p.p-DDE Dieldrin p.p'-DDT trans-Chlordane cis-Chlordane 8 3 2 1 7 5 2 1 27 10 10 3 no 6 192 8 4 100 75 67 70 100 96 98 78 81 Average derived from a total of 100 samples, each sample representing one dozen eggs. The maximum PCB residue was close to 0.03 ppm and the maximum total DDT level (sum of p,p'-DDT and p.p'-DDE levels) was close to 0.3 ppm, although the latter represents a single instance of high p,p'-DT)T and p,p'-DDE levels in the same sample. The second-highest total DDT value was 0.07 ppm. The low levels found in this investigation ought not to be taken as absolute, since recovery studies for eight different egg samples ranged from 25 to 115 percent at the 1-10 ppb level. PCB's, p.p'-DDT and p,p'-DDE, however, had better than 50 percent recovery at all times. Egg samples spiked at the 1 ppm level had >80 percent recovery, except for lindane which had 60 per- cent recovery. The blanks gave no significant findings. The low level of p.p'-DDT and p,p'-DDE in chicken eggs may reflect low pesticide intake by the chickens, since eggs are an important elimination route for these pesticides (//). Table 2 shows a rather even distribution of PCB's and pesticides. The mean levels of p.p'-DDT in the Western and Eastern regions were relatively higher than in the rest of the country only because of one or two indi- vidual high values. The GC patterns of PCB's in individual egg samples were similar to those of Aroclor 1260. During confirma- tion of PCB's on TLC plates two distinct areas were observed which, when scraped and eluted, had two GC patterns consisting of the following Aroclor 1260 peaks numbered according to the system of Reynolds (7) and O.E.C.D. (S): most of: some of: 10 Fraction 1 (Rp,p'-DDE 1-15) 8,9,11, 13, 15,17, If 10,16 Fraction 2 (Rp.p.-DDE 1-00) 10,12,14,16 9 TABLE 2. — Regional distribution of PCB and pesticide residues in Canadian chicken eggs Compound PCB's as Aroclor 1260 Lindane Heptachlor Heptachlor epoxide p.p'-DDE Dieldrin p.p'-DDT trans-Chlordane cis-Chlordane Average ppb ' So: 7 trace trace 1 9 2 12 trace trace z u u m UBS 10 5 4 7 1 2 3 2 S 2 W1 C I traci 8 » Average derived from a total of 20 samples from each region; eac sample represents one dozen eggs. - Trace = < I ppb. The combination of both fractions gave a GC patter quite similar to Aroclor 1260, except for peak 10 whic was considerably higher. The latter may be an indict tion of the presence of Aroclor 1254, but no attemp was made to correct for it. All pesticides reported in the tables were confirmed b TLC as described, including chlordanes whose presenc in eggs has been reported earlier (12). Althoug o,p'-DDT was suspected in several samples and ha been previously reported in eggs (12), this pesticid could not be confirmed by the TLC procedure. Tract (wever, this relationship seemed weakest for PCB's. 7iong eight chemical residues, PCB levels were most often rrelaled with levels of the other seven. Levels of chlori- ted hydrocarbons in wings were correlated with levels in east muscle and in carcass: however, mean levels of cer- n residues differed significantly among wing, muscle, and ^cass even when compared on a lipid basis. effected the closing of the 1970 woodcock hunting season in New Brunswick, Ontario, Canada. At that time a series of 46 woodcock showed a range of 3 to 771 ppm (lipid weight) DDT plus metabolites, with a weighted mean of 60 ppm. In a more comprehensive study in 1973, woodcock averaged 25.8 ppm (lipid weight) DDT and metabolites from 164 analyses of 527 woodcock from New Brunswick (2). In the 1970-71 study of U.S. woodcock, data from analyses were summarized by contaminant and com- pared to residue levels reported elsewhere and to U.S. Government guideline levels for meat destined for hu- man consumption. Analytical findings were put to several other uses: pinpointing of the geographic distribution of woodcock containing mirex; comparison of data from pooled samples versus averages of individually analyzed birds; measurement of north — south geographic varia- tion in residue levels; determination of correlations of residues of various toxicants wtih one another; and quantification of the relationships of residue levels in wing versus breast muscle, and wing versus carcass. Iniruditction om October 1970 to February 1971, 229 woodcock hilohela minor) were collected for chemical analysis im 23 Eastern and Midwestern States. Survey objec- 'es were to establish base residue levels of organochlo- I e insecticides, PCB's, and mercury, and to determine I ; geographic distribution of mirex. Providing impetus !• this survey was an earlier Canadian study (7) which I .S. Dept. of Interior — Fish and Wildlife Service, Paiuxent Wildlife I esearch Center, Laurel, Md. 20811. Analytical Methods Among the 23 States represented in this survey were localities as far north as Maine and as far south as Florida. Several Midwestern States were sampled, in- cluding Minnesota and Missouri. Table 1 lists specific States represented and the number of woodcock col- lected in given counties of each State. Sampling began in October in the Northern States and ended in February in the Southern States. >L. 8, No. 1, June 1974 15 TABLE {.—States and counties sampled for chlorinated hy- drocarbon and mercury residues in woodcock, 1970-71 State County Birds collected Alabama Mobile 10 Arkansas Chicot 2 Faulkner 2 Grant 2 Hempstead 4 Florida Alachua 9 Marion 1 Georgia Cherokee 5 Kentucky McCracken 1 Ohio 4 Louisiana Iberville 16 Pointe Coupee 4 Maine Hancock 3 Penobscot 5 Washington 2 Maryland Calvert Dorchester Montgomery 2 1 1 Worcester 6 Michigan Mackinac 3 Schoolcraft 7 Minnesota Pine 10 Mississippi Clay 5 Leake 5 Oktibbeha 5 Missouri Callaway 4 New Hampshire Merrimack Rockingham 1 1 Strafford 3 Unrecorded locality 5 New Jersey Atlantic 2 Burlington 3 Cape May 8 Monmouth 1 Morris 4 Salem 2 New York Jefferson 8 St. Lawrence 2 North Carolina Johnston 6 Pender Pennsylvania Berks Bradford Centre Erie Tioga Rhode Island Unrecorded locality South Carolina Georgetown Lancaster Tennessee Coffee Hardeman Chester ' Haywood Lauderdale Vermont Essex Caledonia ' Orleans West Virginia Mason 10 Wisconsin Portage 6 Sauk 4 ' Bordering counties: collection sites overlapped. Mercury analyses were performed on 222 of the 229 birds and all 229 were analyzed for chlorinated hydro- carbons. One hundred of these birds were analyzed in five bird pools. Two pools from both Louisiana and New Jersey were analyzed; one pool from each State having a total sample of ten or fifteen birds was an- alyzed. Pool birds were selected randomly from 1 sample from each State. Pools of livers were analyz for mercury and pools of breast muscle were analyz for both mercury and chlorinated hydrocarbons. To determine organochlorine residue levels for win breast muscle, and carcass subsamples, 40 comparis birds were chosen randomly from the 129 woodcc remaining after pooling. Samples of muscle from birds analyzed individually ! chlorinated hydrocarbons consisted of approximati one-half (20 to 30 g) the breast muscle and excluded 1| skin. A 5-g sample was taken from each pooled bi Pooled samples of breast muscle for mercury analy consisted of 1 g of muscle from each bird. Entire livi were used for both individual and pooled samples whi were analyzed for mercury. Each wing sample included both wings, with distal sJ ment and feathers removed. Carcass samples consis I of the remainder of the bird after samples of brej muscle, the liver, wings, skin, gastrointestinal trtj head, feet, and scaled portions of the legs had bi'j removed. All samples were homogenized prior to ari ysis. Analyses were performed by WARF Institu Inc., Madison, Wis. Samples analyzed for chlorinated hydrocarbons wi weighed, air dried with sodium sulfate for 48 hoii extracted with petroleum ether : ethyl ether (17:7) Soxhlet apparatus for 8 hours, cleaned, and separa into two fractions by passage through a florisil colu of petroleum ether : ethyl ether, 95:5, 85:15. An aliq of the first elution was passed through a standardi silicic acid — celite column with petroleum ether, h ane, acetonitrile, and methylene chloride {3). Anal was performed by electron-capture gas chromatogra] on a Barber-Coleman Pesticide Analyzer model 53 The column was glass, 1208 mm by 4 mm, packed v 5 percent DC-200 80/ 100 mesh Gas Chrom Q. Injec temperature was 240°C; column was 200°C; detector was 245 "C. The carrier gas was nitrogen flow rate of 80 ml/min. Lipid weight was delermi from an aliquot of the extract which was reduced dryness on a steam bath and placed in a 40°C o 2-4 hours before weighing. Total mercury content was determined by cold va atomic absorption. Samples were digested by reflu) with sulfuric — nitric acid mixture (4). A mixture hydroxylamine, stannous chloride, and sulfuric acid added to the digest to reduce the mercury II ion! mercury metal. Samples were aerated at 3 liters/ and passed through the absorption cell. Limits of sensitivity (wet weight, ppm) were 0.05 mercury, 0.01 for PCB's, and 0.005 for DDE, D^ DDT, mirex, heptachlor epoxide, and dieldrin. 16 Pesticides Monitoring Jour coveries of mercury from spiked samples ranged from 85 to 98.5 percent. Percentage recoveries for chlori- nated hydrocarbons were: DDE and PCB's, 75-85: DDD and mirex, 80-90; DDT, 75-80: dieldrin, 82-94: and heptachlor epoxide, 85-90. Analytical readings were not corrected for recovery. Confirmation consisted of nmning duplicate analyses. For mercury, five pooled liver samples and three individual livers were duplicated; for chlorinated hydrocarbons, one pooled muscle sam- ple, three individual muscle samples, and four carcasses were duplicated. In addition to arithmetic means, median values are given throughout this paper because all residue data were skewed with most values toward the low end of the distribution. Statistical tests were completed after log]o (x -(- 1) transformation of the data. Trace residues were entered in the computations at the stated "less than" value, and "not detected" values were entered as zeros. Results ami Discussion WOODCOCK AS HUMAN FOOD Data for U.S. woodcock (Table 2) show that residue levels are generally low; average levels of DDT plus metabolites are approximately one-half those of the mean New Brunswick sample reported by Dilworth et al, (2) in 1973. Data are presented by States (Table 3) because concern for residues in game species is centered at the State level. However, sample sizes were small and samples were not selected randomly within States. Therefore, data are statistically representative only of the area(s) I actually sampled and not of the State as a whole. Be- cause woodcock were collected during the hunting season to determine those residue levels to which hunters might be exposed, annual variation by seasons is not measured by these data. Action Guidelines of the United States Department of Agriculture (USDA) for chlorinated hydrocarbons in meat (lipid weight) of domestic animals intended for human consumption are: DDT and metabolites, 7 ppm; PCB's, 5 ppm; dieldrin, 0.3 ppm; heptachlor epoxide, 0.3 ppm; and mirex, 0.1 ppm (John Spaulding, Ph.D., Residue Evaluation and Planning Group, USDA, per- sonal commiinicalion). Tables 2 and 3 reveal that numerous organochlorine means are apparently in excess of the USDA official limits. Because of skewed distributions, medians are both lower than and more representative of the dosage likely to be encountered in any given specimen. Never- theless, many median values also appear to be in excess. In actuality, breast muscle of woodcock contains only 1.9 ± 0.1 percent fat (mean and 1 standard error for the 40 comparison birds), whereas hamburger, for ex- ample, may contain 28 percent fat. or 15 times the mean fat in woodcock. Therefore, in considering the safety of chlorinated hydrocarbon residues in wood- cock (Tables 2, 3), it is appropriate to multiply guide- line levels by approximately 15. There are no median values, including those for mirex which are discussed below, which exceed these adjusted guidelines. The maximum level of mercury in fish muscle allowed by the Food and Drug Administration (FDA), United States Department of Health, Education, and Welfare, is 0.5 ppm (wet weight) (Spaulding, personal communi- cation). Among the State pools of breast muscle analyzed for mercury, the highest residue values, 0.31 ppm for Florida and 0.30 ppm for Alabama, were below the 0.5 ppm level. Mercury levels in breast muscle of woodcock apparently present no hazards to humans according to established tolerance levels. l-OUISIANA WOODCOCK McLane et al. (5) report residues (lipid weight) of heptachlor epoxide, dieldrin, DDE, DDD, and DDT in 33 woodcock collected January 1965 in West Baton Rouge, Pointe Coupee, and Iberville Parishes of Louis- iana. Analyses of 10 birds in the present sample afford a comparison for the same area 6 years later. To make this comparison, the authors' 1970-71 data were re- analyzed according to McLane et al. (5): trace readings were used at one-half the stated "less than" values. This results in data slightly different from Louisiana values in Table 3. The mean (1.87 ppm) and median (0.88 ppm) residues for heptachlor epoxide in 1965 were higher than the mean (0.04 ppm) and median ("not detected") levels found in 1971. The mean (1.65 ppm) and median (0.48 ppm) residues for dieldrin in 1965 do not differ greatly from the mean (0.80 ppm) and median (0.56 ppm) values found in 1971. DDE was present in 1965 at mean (17.90 ppm) and median (16.15 ppm) levels which exceed the mean (6.88 ppm) and median (3.32 ppm) residues of 1971. The 1965 measurements of DDD and DDT included trace amounts of PCB's; therefore, it is not possible to judge whether DDD and DDT have increased or decreased. It would seem that mean (3.65 ppm) and median (3.92 ppm) levels of PCB's in 1971 have increased over the trace amounts present in 1965. GEOGRAPHIC DISTRIBUTION OF MIREX Mirex was found in breast muscle of ten individually analyzed woodcock from five States. Only one of five birds from Maryland had a residue of 0.658 ppm (lipid weight), resulting in a State average of 0.132 ppm. Two of five woodcock from Alabama had residues of 2.18 ppm and 2.20 ppm; State average was 0.876 ppm. Two of five birds from Tennessee had residues of 0.783 ppm and 5.64 ppm: State average was 1.28 ppm. One of ten birds from Louisiana contained 26.7 ppm; State average was 2.67 ppm. Mississippi showed the highest Vol. 8, No. 1, June 1974 17 TABLE 2.— Residues of chlorinated hydrocarbons and mercury in U.S. woodcock. 1970-71 Chemical' DDE Mean Median Range DDT Mean Median Range DDD Mean Median Range Dieldrin Mean Median Range Heptaclilor epoxide Mean Median Range Mirex Mean Median Range PCB's Mean Median Range Mercury Mean Median Range Birds SAMPLED 129 129 129 129 129 129 129 122 Birds with no detectable residue 23 Birds with trace residue 89 119 ■ Mercury analyses are of liver; other analyses are of breast muscle. levels, with four of ten woodcock containing 4.54 ppm, 10.5 ppm, 11.5 ppm. and 33.6 ppm. State average was 6.01 ppm. State pools showed small amounts of mirex in birds killed in Minnesota, 0.714 ppm; Wisconsin, 0.620 ppm; and South Carolina, 0.545 ppm. The only other State pool with mirex was Mississippi, which had 17.7 ppm. Mirex levels up to 0.192 ppm appeared in one carcass sample from each of the following States: North Caro- lina, Kentucky, Tennessee, Florida, Georgia, West Vir- ginia, and Vermont. A woodcock from New York showed 0.141 ppm mirex in the carcass and 1.23 in the wing. In summary, woodcock showed heaviest mirex residues in Mississippi and Louisiana, States where mirex has been used in attempts to control the imported fire ant (Solenopsis saevi,^sima). POOLS VERSUS AVERAGES It is important to determine how well pooled samples reflect averages of individual birds because pooling is Residue level, ppm Wet weight 56 38 63 22 23 12 0.217 0.036 0.004-8.67 0.010 0.005 0-0.220 0.030 0009 0-0.870 0.018 0.005 0-0,550 0.003 0 0-0.082 0.010 0 0-0.440 0,075 0.060 0-0.43 0.197 0.145 0.05-1.1 Lipid weight 11.2 2.47 0.196-432 0.573 0.364 0-14.0 1.64 0.647 0-55.7 1.07 0.381 0-30.7 0.188 0 0-8.67 0.762 0 0-33.6 4.65 3.65 0-25.7 often used to reduce the number and therefore the cos of analyses in surveys of residue levels. The present dat; allow statistical comparison of one series of residue readings, each reading based on five birds pooled prio to homogenization and analysis, with another series o values, each value derived by averaging residue reading from five birds analyzed individually. Pool values ant averages of individuals are paired: one of each is avail abk: from several States. However, geographic variatioi within States has not been completely accounted for the experimental design because the collecting localitie are not completely the same for each pool-average paii Results in Table 4 show a general similarity betwee pools and averages. However, the similarities are nc consistent as shown by the lack of significant correlatio coefficients for DDT and dieldrin. Although t-tests fc paried data revealed no significant differences betwee pairs of logarithmic means for pools and average: means of pools were larger in five of seven cases. more extensive experimentation reveals that poolir increases the levels of residues which are detected, e: planation and quantification will be required. 18 Pesticides Monitoring Journ/ TABLE 3 • — Geographic suwm individual^ ary of chlorinated hydrocarbon and mercui V analyzed U.S. woodcock, 1970-71 Residue levels, ppm ' y residues in State N^ DDE + DDT + DDD DiELDRIN Heptachlor epoxide PCB's Mercury Mean Me- dian Range Mean Me- dian Range Mean Me- dian Range Mean Me- dian Range Mean Me- dian Range Fla. 5 10.6 9.50 3.03-22.1 0.98 1.11 0.34-1,58 2.34 0,89 0-8,67 7.59 6.82 0,82-15.6 0.28 0.28 0,18-0.36 Go. 5 6.75 4.10 2.30-16.8 0.35 0.34 0.30-0.39 0,15 n 0-0.42 6.54 6.58 5.42-7.81 0.17 0.19 0.05-0.23 Ala. 5 110 7.42 5.77-502 1.52 0.59 0.18-5.78 0,08 0 0-0,39 10.9 3.68 2.70-25.7 0.61 0.68 0.29-0.92 Miss. 10 47.2 13.1 1.41-319 0.56 0.42 0.24-1.74 0.14 0 0-0,50 4.62 3.42 0-15.6 0.17 0,16 0,08-0,27 La. 10 8.19 5.12 0.67-33.1 0.87 0.67 0.13-3.41 0.05 0 0-0.38 3.65 3.92 1.38-5.17 0,25 0.25 0,13-0,36 >N.C. 5 9.01 7.15 3.23-15.8 2.48 0.70 0.52-9.62 0.24 0,27 0-0.44 3.86 3.30 2.72-5.43 0,37 0.26 0,05-0.87 5.C. 5 10.2 7.27 5.55-23.7 6.67 0.60 0.31-30.7 0.05 0 0-0.26 4.36 4.13 2,30-7.75 0,57 0.33 0.17-1.10 Tenn. 5 11.3 13.0 3.39-17.0 1.43 1.10 0,40-3,42 0,37 0.32 0.29-0.52 8.25 7.86 4.55-12.5 0.35 0.26 0.20-0.66 \Tk. 5 22.0 8.81 4.21-64.5 4.61 0.67 0.42-13.5 0.22 0 0-0.79 3.03 2.58 2.07-4.13 0.17 0.19 0.06-0.26 Md. 3.71 3.48 2.42-5.76 0.50 0.40 0.20-1.06 0.03 0 0-0.17 5.74 5.71 3.32-8.24 0.22 0,24 0.14-0.28 iV. Va. 1.36 1.60 0.25-2.62 0.19 0.20 0-0,30 0,05 0 0-0.24 3.4! 3.13 1.78-5.67 0.08 0.07 0.05-0.14 Cy. 2.36 1.86 1.12-4.33 1.13 0.44 0.24-2.86 0,17 0.20 0-0.27 4,85 4.88 4.31-5.43 0.09 0.08 0.05-0.14 vlo. 1.95 1.97 1.37-2.50 1.86 0.34 0-6.78 0,11 0 0-0.44 3.40 2,78 1.92-6.13 0.13 0.15 0.05-0.17 U. 15.6 11,7 1.29-47.6 0.88 1.04 0-2.68 0 0 — 8 58 3,38 1.47-19.5 0.13 0.13 0,05-0,22 -l.J. 10 4.14 2.82 0.64-10.2 0.33 0.27 0,20-0.60 0.03 0 0-0.28 2,20 2.12 0-4.77 0.19 0,16 0.08-0.46 'a. 2.64 1.68 1.30-6.00 0.39 0.30 0.20-0.83 0.21 0 0-0.80 2.61 1,88 1.51-5.17 0.10 0.07 0.07-0.17 i4aine 3.43 0.48 0.31-15.2 0.18 0.19 0.13-0.23 0.04 0 0-0.23 2.43 2,60 1.10-3.51 0.07 0.05 0.05-0.12 A. 1.72 0.53 0.44-5.33 0.24 0,22 0.21-0.33 0 0 — 3.64 3,47 2.91-4.63 0.08 0.08 0.05-0.12 J.H. 3.35 2.10 1.05-8.48 0.23 0.24 0-0.37 0.11 0 0-0.29 1.03 1.41 0-2.21 0.09 0.08 0.07-0.12 '(.Y. 2.64 0.89 0.59-6.08 0.32 0.39 0-0.62 0 0 — 6,72 5,00 2,36-14,4 0.15 0.13 0.08-0,23 Hich. 1.43 1.43 0.79-1.86 0.17 0.23 0-0.31 0 0 — 2.45 2,38 1.59-3.68 0.09 0.11 0.05-0,12 Vis. 6.92 2.68 0.39-26.0 0.22 0.25 (M).47 0.14 0 0-0.47 4.83 3,65 1. 51-12. 5 O.IO 0,09 0,05-0.15 linn. 1.79 1,76 0.55-3.10 0.13 0 0-0.53 0.10 0 0-0.53 5.54 3,65 1.71-12.5 0.06 0,06 0,05-0.08 Data for chl N = number Vt.. La., N.H orinated hydrocarbons apply to breast muscle, lipid weight; data for mercury apply to liver wet weight , and^N J° '" '"""''''■ ^'"""'^ "^" '"'' """""'" "'^ ""^ '"' '^^" ""^ ""'"''" '"'l'^'"^'' '" Ihe followmg States: Tenn., Fla., R.I.. TABLE 4.^Conipcuison of pools untl uverufies of residue data for chlorinated hydrocarbons in breast muscle, and mercury in liver, of U.S. woodcock, 1970-71 Residue levels, ppm '■= DDE DDD DDT DiELDRIN Heptachlor EPOXIDE PCB'S Mercury '■Peans Averages Pools 0.808 0.976 0.344 0.400 0.178 0.190 0.258 0,313 0.066 0.082 0,703 0,694 0.079 0.078 anances Averages Pools >rrelaIion 0.2255 0.2644 0.0728 0,0902 0.0201 0,0169 0.0548 0.1031 0.0129 0.0190 0,0357 0,0630 0.0033 0.0030 Coefficients '' 0.630** 1 0 609" 0,303 0,436 0.764* •• 0.579'* 0.946* •• Oata for chio Residue levels ■neans, varian ignificance le rmated hydrocarbons represent lipid weight; data for mercury represent wet weight, in pools and arithmetic averages of mdividual residue levels were transformed by log„, (x + 1 ) prior to calculation of these ces, and correlation coelficienls. Sample size is 20 for chlorinated hydrocarbons and 15 for mercury calculation of these vels: *• = 0.01 > P > 0.001; •" = P < 0.001. 'DL. 8, No. 1, Jul ME 197 4 19 Comparison of sample variances (Table 4) shows that in five of seven cases the pool variance is larger than the average variance. None of these diflferences produce a significant F value when tested. Together they suggest that pool data are less reliable. If this difference is real, it may be because each State average comes from five analyses, whereas each pool comes from a single anal- ysis. However, the mechanism is not obvious. The authors' limited data suggest that pools and averages with a sample size of five are not equivalent and that pool values are larger and more variable. NORTH— SOUTH VARIATION IN RESIDUES Gross inspection of the data indicated that levels of some residues increase from north to south. To examme this relationship the authors assigned each State a value from 1 to 5 as an index of latitude: 1— Florida, Georgia. Alabama, Mississippi, Louisiana; 2— North Carolina, South Carolina, Tennessee, Arkansas; 3— Maryland, West Virginia, Kentucky, Mi.s.souri; 4 — Rhode Island, New Jersey, Pennsylvania; 5 — Maine, Vermont, New Hampshire, New York, Michigan, Wisconsin, Minnesota. These values were then analyzed for correlation with the 23 States' average levels of chlorinated hydrocarbons (breast muscle, lipid weight) and mercury (liver, wet weight); Table 5 shows results. Number of individual birds for each State ranged from 4 to 10 with an average of 5.6 for chlorinated hydrocarbons and 5. .3 for mercury. Table 5 confirms that residues in general are negatively correlated with latitude. Furthermore, except for PCB's, the size of the correlation coefficients is positively correlated with mean residue level (Table 2, wet weight values); hence low levels are probably responsible for the smaller coefficients of DDD through heptachlor epoxide (there were 4 States where no individual muscle samples contained heptachlor epoxide and 1 S where none contained detectable mirex). However, this explanation does not account for the low PCB coefficient. Whereas the insecticides presumably increase toward the South TABLE 5. — Correhiliotis hclwecii avcnif-c residue levels in U.S. woodcock and lalilude inde.x for 23 .Stales of colleclion. 1970-71 ' Correlation Chemical COEFFICIENT = Heptachlor epoxide -0.408 PCB's -0.431' Mirex -0.528' DDT -0.530' Dieldrin -0.543" DDD -0.550" Mercury -0.659"' DDE —0.670"' • Ppm values transformed by logio (x + 1). -' Significance levels: * = 0.05 > P > 0.01; "• = P < 0.001. 20 " = 0.01 > P > 0.001; due to greater agricultural usage, PCB's originate from various sources {6,7) not necessarily related to latitude, and this may explain the weaker relationship. The strong correlation of mercury with latitude (Table 5) presumably reflects greater ambient mercury in wood- cock habitats with decreasing latitude. How the natural and personmade sources of mercury combine to produce such a distribution is not known. CORRELATIONS OF RESIDUES WITH ONE ANOTHER Among the eight chemical residues, levels of PCB's are most often significantly correlated with levels of other chemicals; this is tnie in six of seven cases (Table 6), DDT and its metabolites are closely associated with one another and with dieldrin and PCB's. The absence of heptachlor epoxide and mirex from many samples (Table 2) probably accounts for their lack of correlation with other residues. Mercury, in spite of its relative abundance (Table 2), is significantly associated with only three other materials. However, because mercury is assimilated and stored differently than the chlorinated hydrocarbons, a lesser degree of association is expected. WING RESIDUES AS INDICATORS Wings of woodcock obtained annually from hunters to evaluate the species' population levels and reproductive success ( P > 0.01; *** = P < 0.001. TABLE 7. — Levels of residues in U.S. woodcock wings as indicators of levels in breast muscle and in carcass, 1970-71 Chemical Residue levels, ppm '■- Mean t (PAIRED DATA) Mean difference ±95% confidence interval Correlation coefficient IDE Muscle 0.584 Wing 0.615 1.57 0.031 ± 0.040 0.955*** Carcass 0.640 1.10 0.025 ± 0.046 0.949*** DO Muscle 0.210 Wing 0.115 4.75*** 0.095 It 0.040 0.779*** Carcass 0.172 3.28" 0.056 ± 0.034 0.862* •• DT Muscle 0.114 Wing 0.275 5.44»'* 0.161 ± 0.060 0.492* •• Carcass 0.183 4.92*«» .0.092 ± 0.038 0.839*** :bs Muscle 0.708 Wing 0.593 4.24«»» 0.115 ± 0.055 0.737*** Carcass 0.538 1.80 0.054 ± 0.061 0.730*** ^Idrin Muscle 0.192 Wing 0.199 »M2 0.007 It 0.042 0.937*** Carcass 0.134 1.56 0.064 ± 0.083 0.581*** ptachlor epoxide Muscle 0.075 Wing 0.055 0.961 0.020 ii 0.042 0.626*** Carcass 0.059 0.693 0.004 ± 0.013 0.932*** .esidue levels in ppm lipid weight were transformed by logio (x + I) prior to computations. Sample size is 40. 1 ignificance levels: •• = 0.01 > P > 0.001; «»*=?< 0.001. ■)L. 8, No. 1, Jl NE 1 974 21 LITERATURE CITED (J) Pearce, P. A., and J. C. Baird. 1970. DDT closes New Brunswick woodcock season. Canadian Field-Naturalist 85(1):82. (2) Dilworlh, T. G., P. A. Pearce. and J. V. DobeU. 1973. DDT in New Bninswick woodcock. In manuscript. (3) Armour. J. A., and J. A. Burke. 1970. Method for sepa- rating polychlorinated biphenyls from DDT and its ana- logs. J. Ass. Offic. Anal. Chem. 53{4):761-768. {4) Monk. H. E. (cluiirpcrson). J. A. Pickard. N. A. Smar!. S. H. Yuen, E. W. Alkins. A. J. Beidas, T. E. Burke, H. Crossley. H. Egan. P. W. Lloyd, and E. J. Miller. 1961. Recommended methods of analysis of pesticide residues in foodstuffs. Report by the Joint Mercury Resi- dues Panel. Analyst 86:608-614. f5) McLane, M. A. R.. L. F. Stickel. and J. D. Newsom. 1971. Organochlorine pesticide residues in woodcock, soils, and earthworms in Louisiana, 1965. Pestic. Monit. J. 5(3):248-250. 16) Dustman. E. H.. L. F. Stickel. L. J. Bins. W. J. Reichel, and S. N. IViemeyer. 1971. The occurrence and signifi- cance of polychlorinated biphenyls in the environment. Trans. 36th N. Amer. Wildlife Natur. Resources Conf., pp. 118-133. 17) Risebrough. R.. with L. Brodine. 1970. More letters in the wind. Environment 12(1): 16-27. (8) Clark. E. R. 1971. The status of American woodcock — 1971. U.S. Bureau of Sport Fish, and Wildl., Migr. Bird Popul. Station Admin. Rep., pp. 1-18. 19) McLane. M. A. R.. L. F. Stickel. E. R. Clark, and D. L. Hughes. 1973. Organochlorine residues in woodcock wings, 11 states— 1970-71. Pestic. Monit. J. 7(2):100-103. '>2 Pesticides Monitoring Journa Studies on the Distribution and Flux of Pesticides in Waterways Associated with a Rice fie Id — Marshland Ecosystem ' Thomas M. Ginn " and Frank M. Fisher, Jr.'' ABSIKAC I 'II coastal prairie and niarsliland in Chambers County, Tex., uthors studied the distrihulioii mid flux of chlorinated vdrocarbon pesticides in waterways associated with a rice- fid — Diarshlaml ecosystem. Aldrin applied with .seed rice itered the aquatic ecosystem through drainage of flooded cefields. Chemical alteration of the pesticide was observed; 'eldrin was the primary breakdown product. All insecli- des were distributed unevenly, exhibiting a predilection itr biotic components of the ecosystem. Residue analy.fes of ■presentative species of the aquatic biota indicated signifi- int biological accumulation and passage of these refractory impounds along the food chain. Rapid localization and mceniration of pesticides in living organisms was observed, eproductive tissues exhibited a marlced affinity for the pes- Hdes. Decline of assimilated residues in both biotic and notic components appeared to follow a first-order reac- )n curve. Contamination of the aquatic environment with xaphene during the study period resulted in a ma.ssive II of aquatic organisms. Neither long-term effects nor sig- ficant biological magnification of toxaphene was observed, ther chlorinated hydrocarbon pesticides of unknown origin ?re detected, including DDE. DDD, and DDT. Iniroiluction le toxicity ot chlorinated hydrocarbon pesticides to Huatic organisms has been documented by many vestigators (1-8). Effects of the pesticidal chemicals ; nge from acute intoxication resulting in death of the ganism to more subtle sublethal effects. Though only "he major portion of the research described herein was supported y a grant from The Mood\ Foundation (70-115). Additional sup- ■ort was provided bv a grant from NASA ( V6R44-006-033 ) and 'HS Training Grant 5-TO1-EM-OO025. lowman Gray school of Medicine, Winston-Salem, N. C. 27103. iiology Department. Rice University, Houston. Tex 77001. slightly soluble in water, chlorinated hydrocarbon pesticides enter the aquatic environment dissolved in minute amounts or in greater amounts adsorbed to sus- pended sediment particles (9). Rudd (10) asserted that water is the primary means of residue transport from a treated area to an tmtreated one. The presence in surface waters of such persistent, broadly toxic com- pounds results in exposure of the entire aquatic biota to the residues. Furthermore, in the laboratory, fish and other aquatic organisms have shown a marked ability to accumulate pesticides from the milieu. This has been attributed primarily to two factors: direct absorp- tion of insecticides from water, and assimilation and concentration of residues from food substances. Butler (//) noted the phenomenon of direct absorption of residues by oysters. Chadwick and Brocksen (12) re- ported that accumulation of pesticides by fish was de- pendent on the concentration of pesticides in water. However, Murphy (/.?) asserted that the method of uptake was dependent on the size of the organism. In addition, accumulation of pesticides by organisms in natural aquatic ecosystems has been documented (14-16). This phenomenon of accumulation of pesticides by living organisms appears to be universal (17) and is primarily attributable to the chemical properties of these globally dispersed refractory compounds. The effects of accumulating pesticides are for the most part unknown. Sublethal amounts of these compounds in water have been shown to retard growth (3). decrease reproductive success (18). and alter behavior (19) in aquatic organisms. Most investigations have confined themselves, however, to laboratory studies on isolated 3L. 8, No. I, June 1974 23 components of an aquatic ecosystem or isolated char- acteristics of an insecticide. At present, few data exist on the distribution, locaUzation, and impact of various pesticides in natural aquatic ecosystems. Studies of residues in a few complete ecosystems have been under- taken {16.20). The marshland — estuarine environment is one for which virtually no data exist on the dynamics or short-term effects of chlorinated hydrocarbon pesticides. As the habitat for a great and diverse group of aquatic species, the estuarine environment is a vitally important bio- logical system. The authors" purpose was to investigate the dynamics and distribution of certain chlorinated hydrocarbon pesticides in a marshland — estuarine eco- system. The primary pesticide in the investigation was aldrin applied in cultivation of rice and introduced into the aquatic ecosystem by drainage of flooded ricefields. In addition. 1 1 other chlorinated hydrocarbon pesticides were monitored in the organisms of this aquatic com- munity. Sliuly .Area The study area was composed of approximately 8,000 hectares of coastal prairie and marshland in Chambers County, Tex., adjacent to East Galveston Bay (Fig. 1). The marshlands were used primarily for cattle grazing and the lower prairie was used for rice cultivation. Runoff from the marsh and rice-growing areas passed through a series of personmade drainage canals and natural bayous into East Bay. Salinity of drainage waters varied from 0.5 to 19 parts per thousand (ppt) during the study. Aquatic organisms inhabiting the waters were typically euryhaline — estuarine species. The marshland primarily discussed in this paper was a 300-hectare section of the above area which had been drained: it was cultivated with rice between March and August 1971. Portions of the lowlands not involved in rice cultivation were designated as control areas. Materials and Methods Aquatic organisms were trapped or netted in the main drainage points of treated ricefields and stored at 4''C or quick-frozen ( — 200 until time of extraction and analysis. Small organisms (Palaeinoneles. Brevoortia. etc.) were collected to give a sample size of 30 to 60g. The lower value represented the smallest sample in this study. Routinely, dip nets and seines were used for benthic forms. Cast and gill nets were used for fish and commercial crab traps were used effectively for crabs and some fish. Preparation for extraction was a modification of the method described by A. Wilson of the U.S. Environmental Protection Agency, Gulf Breeze Environmental Research Laboratory, Gulf Breeze, Fla. (personal eommunication). Samples were weighed and 24 FIGURE 1. — Location of ricefield — marshland study are> on Texas coasi placed in mason jars to which anhydrous Na^iSO^ waj added in an exact multiple of the sample weigh- (usually 3 X ). The mason jar was capped with a cuttini assembly (Osterizer Corp.) and the sample was grouni until a homogenous mixture was obtained. The groun sample was frozen (-20 C) for 15 to 20 minutes an reground until a free-flowing powder mixture wej obtained. Repeated freezing and regrinding was ofte necessary to achieve such a powder. The tissue — Na2SC! homogenate was either extracted immediately c wrapped in aluminum foil and stored in a freezil (- 20''C). Analysis was of whole-body residues excej when an organism was too large. In such case representative tissues were analyzed, as indicated in tl text and tables. Pesticide residues were extracted in Soxhlet extracto with petroleum ether as described by A. Wilson (person eommunication). A weighed quantity of tissue-desicca homogenate equivalent to approximately 30 g tissue w extracted for 4 hours with 250 ml petroleum eth (Nanograde, Mollinkrodt) at a temperature high enou; to produce cycling of the solvent every 5-10 minut (approximately 90"C). Following extraction, the petr leum ether was reduced to approximately 15 ml ov steam using a three-ball Snyder column and was quan tatively transferred to a separatory funnel for partitic ing over acetonitrile. The petroleum ether fraction w adjusted to 25 ml and partitioned twice over 50 acetonitrile. Each time the mixture was manua agitated for 1 minute and allowed to separate. T acetonitrile was collected in a crystallizing dish a evaporated just to dryness on a slide warmer (40''< Pesticides Monitoring JouR^ Prolonged drying or excess heating was avoided to prevent loss of pesticides due to volatilization. The residue was resuspended in petroleum elher and subjected to column chromatography on tlorisil (Floridin Co., Berkeley Springs, W. Va.). A two-fraction prepara- tive chromatographic separation involved transfer of the residue to 10 g of florisil in a 400-by-20-mm glass chromatography column. Elution was effected with 150 ml of a 6-percent solution of anhydrous ethyl ether in petroleum ether followed by elution with 150 ml of a 15-percent solution of ethyl ether in petroleum ether. The florisil required heating at 120°-135°C for at least 5 hours before use. This procedure separated dieldrin and endrin from other chlorinated pesticide residues (.?/). The 6-percent eluate was reduced to suitable volume over steam and subjected to gas-liquid chromatography (GLC). The 15-percent eluate required further prepara- tion. The fraction, reduced to approximately 20 ml over steam, was quantitatively transferred to a 400-by-20-mm glass chromatography tube containing 10 g of a 1:1 by weight mixture on magnesium oxide and Celite 545 (Johns Manville Co.). Elution was effected with 100 ml petroleum ether over vacuum sufficient to produce an elution rate of approximately .^5 ml/min. The eluate was reduced to suitable volume, quantitatively trans- ferred to a graduate cylinder, and adjusted to a known volume with petroleum ether prior to analysis by GLC. 'Pesticide residues were recovered from core samples (2.5 by 15 cm) of soil by triple extraction with aceto- nitrile. Extraction was performed with wet soils to maximize recovery {22). A portion of each sample was retained and dried to constant weight at 130°C for determination of weight. Samples weighing 250-300 g were placed in mason jars with 100 ml acetonitrile and mixed for 2 minutes in a high-speed blender. After the soil had settled the acetonitrile was decanted into an evaporating dish through a funnel containing anhy- drous Na^SOj to absorb water. Extraction was repeated 1 second and a third time using 100 ml acetonitrile and slending for 2 minutes. Following decantation of the bird extract, the soil was poured into the funnel, rinsed •vith acetonitrile, and discarded. The acetonitrile was ;vaporated to dryness on a slide warmer. The residue vas resuspended in petroleum ether and subjected to a wo-fraction purification on florisil. In this case, botii he 6-percent and 15-percent fractions were subjected no GLC without further purification. 3ip samples of water were collected in I -gal. brown jlass bottles and refrigerated until time of extraction. ^11 analyses were completed within 30 hours after ollection. Samples of 1,500 ml were extracted in a eparatory funnel by shaking for 5 minutes with 00-150 ml petroleum ether. The water and petroleum ther were allowed to separate and the petroleum ether vas collected in an Erienmeyer flask. Extraction was repeated a second and a third time. The aqueous phase was discarded and the petroleum ether phase was dehydrated with anhydrous Na^SO,. The extract was reduced to a known volume over steam. No further purification of this extract was necessary prior to analysis by GLC. This methodology gave 100-percent recovery of dieldrin from spiked samples. Salinity esti- mates were accomplished with standard hydrometers measuring density ranges from 1.000 to 1.030, or by chloride measurement using ampimetric titration with a chloridometer. Residue analysis was by GLC with a Varian Aerograph Model 2100 dual-channel chromatograph equipped with tritium electron-capture detectors. Detection by electron capture allowed quantitation of organochlorine insecti- cides at amounts ranging from 10 " g for lindane to 1 0-1" g for p.p'-DDT. Readout was on a Varian Aero- graph Model 20 dual-pen recorder. On-column injection into an all-glass system was incorporated to prevent decomposition of certain organohalogen insecticides (25). Columns were 6-foot capillary U-tubes with 2-mm internal diameter. Packing material consisted of 80/100 mesh Gas Chrom Q (Applied Science Laboratories, State College. Pa.) solid phase with silicone oil liquid phases including 3 percent DC-200, 5 percent QF-1, a 2:1 mixture of 5 percent QF-1 and 5 percent DC-200, and a 3:1 mixture of 3 percent DC-200 and 10 percent OV-17. Nitrogen was used as carrier gas at 65 psi and rate (soap film flowmeter) of 40 ml.'min. During operation the column temperature was maintained at 190°C. The injector was maintained at 215 C. Sufficient sensitivity was achieved by operating the tritium detectors at 2I5°C. The detectors operated on a 90-volt direct-current mode. Though linear range of electron-capture detectors is limited (24). linearity was obtainable in the range of pesticide concentrations assayed (lO" to lO-a g). Quantitation of data was based on comparison of printout of sample extract peak height with the peak heights of standard solutions of pesticides. Standards were in- jected following every third sample extract. Standard solutions were made using analytical grade chlorinated hydrocarbon pesticides obtained from the Pesticide Repository, Perrine, Fla. (See Appendix for common and systematic names of pesticides used in analysis.) Analysis was .confirmed by using two liquid phases of difl'erent polarity such as pC-200 and QF-1. Another method employed binary solvent systems as described by Bowman and Beroza (25); this relies on solubility ratios of pesticides in immiscible solvents. Results Chlorinated hydrocarbon pesticides entered the aquatic ecosystem in the form of aldrin applied as a dressing on /oi. 8, No. 1, June 1974 25 seed rice (0.28 kg aldrin/ hectare (ha.)). The cultivation process involved flooding ricefields to which aldrin- treated seed rice had been applied aerially, followed in 24-48 hours by discharging flood waters into drainage canals. Residue analyses of drainage canal water samples and ricefield soil samples taken before and after pesticide application are presented in Tables 1 and 2. respectively. Aldrin had been applied to the field; yet dieldrin, rather than aldrin, was the primary residue detected. This was not unexpected since dieldrin. a persistent chlorinated hydrocarbon pesticide in its own right, is the epoxida- tion product of aldrin. The epoxidation process was doubtlessly hastened by environmental conditions of the ricefields, including elevated temperatures, high moisture levels, pH. Eh, and soil structure (26,27). Concentrations of aldrin and dieldrin in water were extremely small. This is impressive, considering that these residues were the primary source of aldrin and TABLE I. — Pesticide residues delected in water from rice- field drainage canals before and after pesticide application Time. WK 1 Residue. PPB (MG/l)- Aldrin Dieldrin -2 ND ND -1 ND ND 0.5 0.270 0.440 4 ND 0.172 8 ND 0.062 12 ND 0.040 13 ND 0.023 14 ND 0.034 15 ND 0.031 Represenls no. of weeks before ( — ) or after planting of aldrin- treated seed rice. ND = none detected ( •^ 0..M) OX; ^ S> a. a z a. 0 20 \ _i Ol Q 0.10 ♦-• 5 0 5 10 15 WEEKS BEFORE/AFTER APPLICATION FIGURE 2. — Dieldrin residues in ricefield drainage walei before and after planting of aldrin-treated rice 26 Pesticides Monitoring Journa Though 1 1 additional chlorinated hydrocarbon pesticides were monitored (chlordane, p,p'-DDD. p.p'-DDE. p.p'-DDT. endrin heptachior, heptachlor epoxide, lin- dane, methoxychior, mirex, toxaphene), only toxaphene was detectable in water samples during the study period. Because of the chemical properties of toxaphene, it was not possible to quantitate the data obtained. Toxaphene residues and their effects on aquatic biota are mentioned later in this section. In the control marshland drainage system, water samples were taken regularly during the study period. No chlorinated hydrocarbon pesticide ■esidues were detected. Soth aldrin and dieldrin were accumulated in soil (Table I). The rapid decline in aldrin concentration was prob- ibly caused by loss through volatilization and decompo- lition: the rapid increase in dieldrin during the first ■\ weeks presumably was caused by significant epoxi- iation of aldrin to dieldrin. The rate of loss of dieldrin rom soil following the fourth week appeared to be pro- )or(ional to the amount of residue in the soil. This vould conform to Kearney et al. (28). who reported that loss of pesticides from soils followed a first-order reac- ion. The possibility of pesticide translocation by trans- ported particulates has been suggested by Keith (9). i 'hough the data are incomplete, pesticide analysis of ■■ottom sediments from the ricefield drainage canal in- dicated localization of dieldrin of the same order of Magnitude as that in ricefield soil (Table 3). 'ABLE 'i.— Pesticide residues detected in drainage canal bottom sediments before and after pesticide application Time, Residue, ppb (;/g/l)=.2 WK ■ Aldrin Dieldrin —4 ND ND -2 ND ND 2 ND 5.4 8 ND 2.4 15 ND ND Samples analyzed: composite core 2.5 bv 15 cm. dry weight ND = none detected (n]y while the chemical was present in the surrounding vater. Dieldrin residue levels in L. inacrochinis. how- ;ver, more closely resembled those of Lepisosleiis, reach- "Hg a peak after 4 weeks and slowly declining there- ifter. DDT and metabolites were present in several L. nacrochinis. but at levels much lower than in Lepi- osteiis. PABLE 4. — C Molina! cd hydrocarbon pesticides delecwd in polled gar (Lepisosteiis oculatus) after pesticide apph'calion Time, Residue PPB (/jg/kg )=,:•. WK ' Aldrin Dieldrin DDE DDT DDD 0 7.4 4.9 368.0 21.7 57.6 0 28.4 11.9 524.0 172.0 198.0 14.7 9.6 113.0 68.0 82.1 5.7 .11.4 88.0 trace 29.0 .1,9 96.8 58.0 14.2 19.3 7.1 153.0 107.0 34.7 79,7 16.3 275.0 145,0 42,2 101.0 12.1 198.0 82.7 5,9 14,3 2.6 110.0 59.0 ND 6,4 15.1 209.0 181.0 22,2 65,1 9 ND 98.0 191.0 ND ND 14 ND 69.5 87.5 - - Represents no, of weeks after planting of aldrin-treatcd seed rice 0 = week of planting. Samples analyzed: 1:1 mixture of liver and muscle, wet weight ND = none detected (<(/g/kgl, REVOORTIA fighest recorded levels of aldrin and dieldrin in quatic organisms were in Brevooriia sp. Assimilation of esticide was rapid: extremely high levels were reached uring the first day of exposure. This is understandable 1 view of the findings of Murphy (/.?) concerning up- ike of residues by fish. In contrast to large fish, small 4h assimilate residues primarily by absorption from ater rather than by consumption of any foods. Bie- lonia collected were in the 10-40-mm range and •obably absorbed the pesticide directly from the water. s a result, pesticides were assimilated with exceptional ficiency and little time lag. IVERSE FISH ;sticide data from the diverse fish listed in Table 5 e significant from the standpoint of survey. Dieldrin sidues were observed in all fish collected after aldrin id been introduced into the habitat, and the residues persisted for a substantial length of time. These fish represent food .sources of many organisms of higher trophic levels. As mentioned previously, residues of toxaphene were detected in waters of the study area. This was the re- sult of contamination of the drainage canals during aerial application of an unknown concentration of toxaphene to a ricefield in the study area as treatment for gras.s- hopper infestation. The application of toxaphene was discovered following residue analysis of fish carcasses sampled from a massive fish kill which followed the spraying. The extreme sensitivity of fish to toxaphene (5) and the unusually high solubility of toxaphene in water (.^ ppm) were factors which indicated acute toxa- phene poisoning as the source of the fish kill. The num- ber of fish killed was estimated in the tens of thousands and included catfish, menhaden, bluegill, carp, mullet, and numerous species of minnows and fry. The popu- lation of grass shrimp was so reduced after the toxa- phene application that no further sampling of the or- ganism was possible for the following 6 weeks. Because of the nature of to.xaphene, residue levels could not be quantitated. Only minute quantities of toxaphene were detected in the water following the fish kill. Fur- thermore, the limited persistence of toxaphene pre- vented accumulation of residues in most organisms of the ecosystem. Residues were detected in several aquatic feeding birds (Louisiana heron: Hydranassa tricolor: lesser yellow legs: Toianiis fiavipes: and black skimmer: Rynchops nigra), in two species of live fish (gar, and flounder: Piatichthys flesus). and in certain aquatic insects (dytiscids). Blue crabs sampled contained no detectable residues, nor were toxaphene residues de- tected in any other terrestrial organisms of the area. The significance of the input of toxaphene into the ecosystem was diflicult to evaluate. Reduction of orga- nisms was temporary and repopulation occurred within days or weeks. No long-term changes in species com- position and diversity were noted. The lack of persist- ence of the pesticide prevented significant biological accumulation of the residue. RE.SIDUES AND REPRODUCTION The increased metabolic activity accompanying repro- duction and the high lipid content of eggs would favor accumulation or deposition of pesticides in both repro- ductive organs and eggs. The deposition of pesticides in reproductive materials could have an important effect on the reproductive success of an organism and ulti- mately could afl'ect the natural history of the species. Levels of pesticides were determined for body tissues and eggs of two Lepisosteiis oculatus collected in the study area and are reported in Table 6. In the case of nearly every residue present, a tenfold increase in con- OL. 8, No. 1, June 1974 29 TABLE 5. — Pesticide residues present in bliiegilt iLepomis macrochirus), menhaden (Brevoortia sp.), and other species of fish before and after pesticide application Organism Residue, ppb {iig/kg)-'' WK 1 Aldrin DlELDRIN DDE DDT DDD -20 L. Macrochirus ND 7.7 ND ND ND -10 ■■ ND 34.0 12.0 ND ND 0 " 24.5 92.0 39.8 13.3 17.4 0 " 28.6 45.0 21.6 ND 7.5 4 ND 410.0 30.8 ND ND 10 ■• ND 64.6 — - - 15 " ND 7.3 ND ND ND -20 Brevoortia sp. ND 4.7 5.7 ND ND 0 550.0 1380.0 25.2 ND ND 0 830.0 2200.0 29.5 ND ND 8 •• ND 81.4 ND ND ND 8 •• 6.1 160.0 16.9 ND ND 10 " ND 80.0 13.8 ND ND II ND 32.5 - - - 2 Micropogon uiidu- tatus (croaker) ND 2.9 33.6 ND ND 8 Syngnalhus sp. (pipefish) 46.8 555.0 48.6 ND ND 12 Mugil sp. (mullet) ND 104.0 15.2 ND ND 13 ND 53.0 - - - 13 Iclahirtts pufu- talus (catfish) — 107,0 — — — 13 Plalichtliys flesus (floutider) - 235.0 - - - ' Represents no. of weeks before ( — ) or after planting of aldrin- treated seed rice. -' Samples analyzed: whole body, wet weight. ■' ND = none detected (f chemically active compounds. Studies of long-term ffects of small, sublethal doses of pesticide would seem o be the most logical method for evaluating the ultimate fl^ect of chlorinated hydrocarbon pesticides on living ystems. A cknowledgments "he authors wish to thank the Kinetics International )ivision of Ling-Temco-Vought, Inc., Tyler, Tex., for le gift of an all-terrain vehicle used in these studies, appreciation is also extended to Thomas W. Duke, h.D.. Director of the U.S. Environmental Protection .gency Laboratory, Gulf Breeze, Fla.. and Lyle Wong, Ph.D., research assistant, for reading this manuscript. Special thanks are due The Brown Foundation, Mrs. G. R. Canada. Mrs. Hubert Curlee. Charles Ezer, Joe Lagow, Ralph Leggett, William L. Moody IV, and J. Whitehead, who generously permitted us to use the study area described in this report. LITERAFURE CITED {!) Anderson. B. G. 1959. The to.xicity of organic insecti- cides to Dapluiici. In Biological Problems in Water Pnl- iiilion. Robert A. Taft Sanitary Engineering Center. Cincinnati. Ohio. pp. 94-95. (2) Butler, P. A. 1962. Effects on commercial fisheries. In Effects of Pesticide on Fish and Wildlife in 1960. Bu- reau of Sport Fisheries and Wildlife Circular 143. Fish and Wildlife Service. U.S. Depl. of Interior. (3) Butler, P. A. 1969. The significance of DDT residues in estuarine fauna. In Chemical Fallout. (Miller and Berg. ed.). Chas. C. Thomas Publisher. Springfield. 111. (4) Graham, R. J. 1959. Effects of forest insect spraying on trout and aquatic insects in some Montana streams. In Biological Problems in Water Pollution. Robert A. Taft Sanitary Engineering Center. Cincinnati, Ohio. pp. 62-65. (5) Henderson. C. Q. H. Picliering, and C. M. Tarzwell. 1959. The toxicity of organic phosphorus and chlori- nated hydrocarbon insecticides to fish. In Biological Problems in Water Pollution. Robert A. Taft Sanitary Engineering Center. Cincinnati, Ohio. pp. 76-88. (6) Loosanoff, V. L. 1959. Some effects of pesticides on marine arthropods and mollusks. In Biological Prob- lems in Water Pollution. Robert A. Taft Sanitary Engi- neering Center. Cincinnati. Ohio. pp. 89-93. (7) Nicliolson. H. P. 1967. Pesticide pollution control. Science 158:871-876. (8) Wursler, C. F.. Jr. 1968. DDT reduces photosynthesis by marine phytoplankton. Science 159:1474-1475. (9) Keith. J. A. 1966. Reproduction in a population of her- ring gulls (Lams argentatiis) contaminated by DDT. J. Appl. Ecol. 3 (Suppl.):57-70. (10) Rudd, R. L. 1964. Pesticides and the Living Landscape. U. of Wis. Press. Madison. Wis. (//) Butter. P. A. 1966. Pesticides in the marine environ- ment. J. Appl. Ecol. 3 (Suppl.):253-259. 112) Ctuutwick, G. G., and R. W. Brocksen. 1969. Accu- mulation of dieldrin by fish and selected fish-food or- ganisms. J. Wildl. Manage. 33(3):693-700. tl3) Murphy, P. G. 1971. The effect of size on the uptake of DDT from water by fish. Bull. Environ. Contam. To.xicol. 6:38-45. {14) Hunt, E. G., and A. I. Bischoff. 1960. Inimical effects on wildlife of periodic DDD application to Clear Lake. California Fish and Game 46:91-106. (15) Hunt. E. G. 1966. Biological magnification of pesti- cides. In Scientific Aspects of Pest Control. Nat. Acad. Sci. Nat. Res. Counc. Publ. 1402. Washington, D.C. pp. 251-262. (16) Woodwell, G. M., C. F. Wurster, Jr., and P. A. Isaac- son. 1967. DDT residues in an east coast estuary: a case of biological concentration of a persistent insec- ticide. Science 156:821-823. (17) George, J. L., and D. E. H. Frear. 1966. Pesticides in the Anarctic. J. Appl. Ecol. 3 (Suppl.):155-167. (18) Grosch, D. S. 1967. Poisoning with DDT: Effect on reproductive performance of Artemia. Science 155- 592-593. 'OL. 8, No. 1, June 1974 31 (19) Cairns, J., N. R. Foster, and J. J. Loos. 1967. Effects of sublethal concentrations of dieldrin on populations of guppies. Proc. Acad. Natur. Sci. Philadelphia 119: 75-91. (20) Meeks, R. L. 1968. The accumulation of "CI ring- labeled DDT in a freshwater marsh. J. Wildl. Manage. 32:376-398. (21) Johnson, L. Y. 1962. Separation of dieldrin and endnn from other chlorinated pesticide residues. J. Ass. Offic. Agr. Chem. 45:363. (22) Saha, J. G. 1971. Comparison of several solvents for extracting root absorbed radioactive dieldrin from wheat. J. Econ. Entomol. 64(l);50-53. (23) BoneUi, E. ].. and K. P. Dimick. 1964. Gas chroma- tography and electron capture for analysis of pesticides. In Lectures on Gas Chromatography. (Mattick and Szymanski, ed.). Plenum Press, New York. (24) Cieph'nski. E. W. 1964. Instrumental aspects of pesti- cide analysis by gas chromatography. In Lectures on Gas Chromatography. (Mattick and Szymanski, ed.). Plenum Press, New York. (25) Bowman, M. C, and M. Bcroza. 1965. Extraction p-values of pesticides and related compounds in six binary solvent systems. I. Ass. Offic. Anal. Chem. 48(5):943-954. (26) Harris, C. R., and E. P. Lichtenstein. 1961. Factors affecting the volatilization of insecticides from soils. J. Econ. Entomol. 54:1038-1045. (27) Kiigemagi, U., H. E. Morrison, J. E. Roberts, and W. B. Bollcn. 1958. Biological and chemical studies on the decline of soil insecticides. J. Econ. Entomol. 51: 198-204. (28) Kearney. P. C. R. G. Nash, and A. R. Iscnsee. 1969. Persistence of pesticide residues in soils. In Chemical Fallout. (Miller and Berg, ed.). Chas. C. Thomas Pub- lisher. Springfield, 111. pp. 54-67. (29) Bonn. J. M.. T. J. DeCino, N. W. Earle, and Y. Sun. 1956. The fate of aldrin and dieldrin in the animal body. J. Agr. Food Chem. 4:937-941. (30) Moore. N. W. 1967. A synopsis of the pesticide prob- lem. Advan. Ecol. Res. 4:75-125. 32 Pesticides Monitoring Journ Selected Chlorinated Hydrocarbons in Bottom Material from Streams Tributary to San Francisco Bay' LeRoy M. Law and Donald F. Goerlitz ABSTRACT iis pari of a study of llie einironmenlal quality of San ■'rancisco Bay, bottom material from 26 streams tributary 10 the Bay were analyzed for clilordane. DDD. DDE, DDT. »nd PCB residues. These compounds were present in essen- ially all streams tested. Chlordane proved to be ubiquitous. •ith a concentration range similar to that of the other com- ounds. Noteworthy was the occurrence in one stream of .olychlorinatcd naphthalene residues. Compounds occurring n concentrations above 20 /tg/kg were identified in most nistances by combined gas chromatography / muss spectrom- try. Introduction an Francisco Bay is one of the principal aquatic re- jurces of the State of California. Millions of people re- ding on or near the shores of the Bay utilize its waters )r recreation, waste disposal, mining of commercially iluable salts and minerals, marine navigation, fishing, Md general aesthetic enjoyment. The Bay receives wa- 'Ts from many sources, large and small, which drain densely urbanized area developed agriculturally and dustrially. In order to assess the potential contamina- i3n of the Bay from chlorinated hydrocarbon com- Dunds, the U.S. Department of the Interior — Geologi- ! Survey initiated a study in February 1972 to obtain 'ickground information. Of special interest was the iput attributable to the numerous streams that dis- arge into San Francisco Bav. J.S. Department of the Interior— Geological .Survey, Menlo Par]< ..alif. DL. 8, No. 1, June 1974 The worldwide distribution of residues of DDT and PCB's (polychlorinated biphenyls) is well established (1-11). Furthermore it has been demonstrated (12) that PCB's are accumulated and concentrated in certain or- ganisms by as much us five orders of magnitude greater than the organism's aquatic environment. Wood- well and coworkers (13) have studied the biological concentration of DDT residues in organisms of increas- ing trophic levels and found it to be more than three orders of magnitude. Most chlorinated hydrocarbon pesticides are only slightly soluble in water and usually enter the hydrologic environment sorbed onto partic- ulate matter (14). Even when dispersed in aqueous media they sorb onto sediment (15). For example, Bevenue et al. (16) found that the ratio of chlorinated pesticides associated with sediment to that in solution was 9,000 to 1. PCB's, because of their similarity in chemical properties to some insecticides, can be expected to behave in a water/ sediment mixture in a like manner. Depending upon flow conditions sorbed compounds may be either in transport or deposited as part of the stream- bed matrix. Hence it was decided to survey the numerous Bay-area streams by analyzing the bottom material. Nisbet and Sarofim (17) have estimated that of the 5 x 10'' tons of PCB's produced in the United States during 1930-70, approximately 3.9 x lO-"* tons, or 78 percent, have been lost to the environment. Riseborough and de Lappe (18). in discussing the mass balance of pollu- tants, state that in the last 15 years, 6 x lOi'^ g (6.6 x 10^ tons) per year of DDT have been manufactured in the United States for both domestic use and export. Because DDT is intentionally applied to the earth, vir- 33 tually all of it has been lost to the environment, whereas PCB's are leaked to the environment by usage. In view of these large production figures the chlorinated hydro- carbon compounds DDT and PCB were considered the most likely pollutants to be found. Therefore, these compounds were chosen for study and this report sum- marizes the findings. Sampling and Analytical Procedure In February and March 1972, bottom material from 26 streams that discharge into San Francisco Bay was col- lected for analysis. Sampling sites are shown in Figure 1. The bottom material was collected in wide-mouth glass bottles equipped with Teflon-lined screw-top clos- ures. Samples of soft bottom material were obtained by scooping directly with the glass bottle. When the tex- ture of the bottom material prevented this method, samples were obtained by scooping the material with a chromium-plated garden trowel and then placing the sample in a bottle. All implements and bottles had been freed of any contaminants that would interfere with analysis. Upon receipt in the laboratory the samples were wet- sieved through a screen with a 2-mm-pore diameter to isolate the material most likely to be transported during high flow. The immersed-screen method described by Guy (19) was followed, using water collected with the FIGURE 1. — Map of San Francisco Bay area sliowing stream bed sites sampled for chlorinated hydrocarbons 34 bed material instead of distilled water. The liquid phase was separated from the solid material that passed through the screen by allowing the mixture to stand for several hours and decanting as much water as possible. Finally, the sieved sample was stored at 4°C to inhibit microbial degradation until extraction was initiated. The procedure used in this study was a slightly modified version of the Geological Survey method for analyzing aquatic sediments for insecticides (20). A 25-g subsample of the wet bottom material was placed in a glass-stop- pered flask and shaken for 20 minutes with 20-m] redis- tilled pesticide-grade acetone. Fifty ml of redistilled pesticide-grade hexane was added to the acetone/ bottom- material slurry, and shaken for an additional 5-10 minutes. The extractant was decanted into a separatory funnel containing 500 ml of organic free distilled water. In a similar manner the extraction was repeated two • more times, the only ditTerence being that the volume of acetone used was reduced from 20 ml to 10 ml. The combined extracts were washed with three 500-ml vol- umes of distilled water and placed over anhydrous sodium sulfate to dry. After drying, the solvent volume was reduced to a few milliliters using a Kuderna-Danish evaporative concentrator and finally to a few tenths of a milliliter by evaporation under a stream of warm dry nitrogen. Mean recovery for the insecticides DDD, DDE, and DDT was 92. 99, and 99 percent, respectively. Mean recovery for PCB was 100 percent. The compounds were initially isolated from interfering coextractives by adsorption chromatography. The con- centrated extract was placed on a l-by-8-cm column of alumina deactivated 9 percent by weight with water and eluted with 20 ml of hexane. The eluate volume was reduced to a few tenths of a milliliter and transferred to a second l-by-8-cm chromatographic column of silica gel deactivated 3 percent by weight with water. Sufficient hexane was added to the column to total exactly 25 ml of eluate. At this point the elution solvent was changed to benzene and an additional 10 ml of eluate was col- lected separately. The PCB's were thereby isolated in the first fraction collected from the silica-gel column, and the DDT family and chlordane were in the second fraction. Finally the extracts were treated with metallic mercury, following the procedure of Goerlitz and Law (2/), forf the removal of sulfur interferences. The purified extractsi were subjected to gas-chromatographic analysis for iden- tification and quantitation of components. Two Variam Aerograph Model 600D gas chromatographs were used. The instruments were fitted with concentric-type tritium* foil electron-capture detectors and connected to Honey- well recorders through an Infotronics Model CRS 100' electronic integrator. One chromatograph was equipped with a glass column 1.8 m long by 1.8 mm i.d., packed with 60-80-mesh Chromosorb W-HP coated with OV-1 3 percent by weight. The other chromatograph was Pesticides Monitoring Journal equipped with a glass column of the same dimensions, packed with 100-120-mesh Supelcoport coated with OV-17 1.5 percent by weight and QF-1 1.95 percent by weight. Operating conditions for both chromatographs were: oven temperature, 195°C; inlet temperature, 210° C; and nitrogen carrier gas flow rate, 30 ml/min. The DDT family of pesticides were measured by com- paring the area under the chromatographic peaks with the area of appropriate standards. Multiple peak resi- dues such as chlordane and PCB's were quantitatively measured by comparing the sum of the areas of the two most prominent peaks with the appropriate analyti- cal standards. The approximate minimum detectable concentrations of the compounds reported, employing the stated analytical conditions, are as follows: DDE: 0.05 X 10-6 ^g/kg; chlordane, DDD, o,p'-DDT. and p.^-DDT: 0.1 X 10-'' fjig/kg: and PCB: 1 x IQ-o fig/kg. When the concentration of any component of interest was great enough, combined gas chromatography/ mass spectrometry was used to confirm the identity of the compound. A Finnigan Instrument Corporation Model 1015 quadrupole mass spectrometer and Systems Indus- tries System 150 computerized controller and data out- put system were used. The mass spectrometer was connected to a Varian Aerograph Model 1700 gas chro- matograph via a glass jet separator. The gas chromato- graph was equipped with a glass column 1.2 m by 2 mm i.d., packed with Gas Chrom Q coated with OV-1 3 percent by weight. The temperature of the column oven was programmed at a rate of 6-C/min from 175" to 235°C and then held isothermally until completion of analysis. Helium was used as a carrier gas at a flow rate of 30 ml/min. Results and Discussion Results of the analysis of bottom material samples from San Francisco Bay Area streams are given in Table 1. Residue concentrations, uncorrected for percent recovery, are expressed as /xg/kg based on the oven-dried weight of the bottom material, determined on a separate sub- sample. Site numbers referred to in Table 1 are the numbered sampling sites on the map in Figure 1. All residue identities in Table 1 were established by elution order when the sample extracts were subjected to column chromatography and by peak matching when analyzed by electron-capture gas chromatography. When concen- tration values were great enough, a gas chromatograph/ mass spectrometer (GC/MS) was used to confirm the identity of a residue. Those residues that were confirmed by GC/MS are noted in Table 1. Because this study was exploratory in design and scope, quantitative conclusions should not be drawn; however, certain generalizations can be observed. Chlorinated hydrocarbon residues were found in all stream bed sam- ples analyzed, thus illustrating their widespread distri- VoL. 8, No. 1, June 1974 TABLE 1. — Chlorinated hydrocarbon residues found in San Francisco Bay Area streams Residue concentration in im/kg '■■- Site No. Stream ul z ■< a § X U Q Q D u Q Q Q "o. 9 a U a. 1 Colma Creek 39 3.0 2.0 1.7 3.2 3.9 2 Colma Creek 19 4.1 1.8 0.80 5.3 12 3 Belmont Creek 660 41 17 89 200 52 4 Cordilleros Creek 20 3.8 3.5 4.0 13 14 5 Cordilleros Creek 33 11 6.1 5.2 39 6.0 6 Redwood Creek 40 8.4 52 4.2 24 25 7 San Francisquito Creek 7.1 2 2 2.1 0.96 7.1 1.2 8 San Francisquito Creek 9 Los Trancos Creek 670 21 160 10 43 5.5 20 1.3 150 5.6 430 21 10 Stevens Creek 7.8 0.0 0.0 0.0 12.3 180 1 1 Stevens Creek 190 19 22 II 33 30 12 Los Gatos Creek 0.0 1.8 0.87 0.28 2.2 0.0 13 Los Gatos Creek 280 33 25 11 32 170 14 Guadalupe River 17 3.5 2.3 0.87 3.2 (=) 15 Guadalupe River 9.6 3.1 1.8 0.52 1.6 2.7 16 Alamitos Creek 46 18 26 2 2 14 610 17 Coyote Creek 83 86 i2. 5.7 31 14 18 Coyote Creek 7j_ 17 II 2.6 4.3 12 19 Alameda Creek 13 5.6 4.5 0.16 0.57 11 20 Alameda Creek 52 12 9.2 2.4 2.1 30 21 Arroyo de la Laguna 200 27 23 4.5 7.3 160 22 Arroyo de la Laguna 22 3.5 4.3 0.77 1.3 n 23 San Lorenzo Creek 15 7.0 7.5 0.0 1.7 25 24 Wildcat Creek 87 18 4.1 2.7 6.5 21 25 Wildcat Creek 45 8.8 6.1 1.8 9.7 43 26 San Pablo Creek 65 2.0 3.4 2.0 3.6 27 27 Union Creek 200 45 16 2.6 2.8 140 28 Green Valley Creek 0.0 1.9 2.3 0.36 0.86 5.3 29 Napa River 10 2 2 2.7 0.78 2.3 8.8 30 Napa River 0.0 46 0.0 9.7 73 1400 31 Napa River 97 16 11 2.8 8.4 7.6 32 Sanoma Creek 4.3 0.98 1.0 0.25 19 5.0 33 Petaluma River 130 8.3 5.5 3.8 2.7 27 34 Navato Creek 62 3.5 3.6 2.4 10 10 35 Miller Creek 3H) 16 11 13 8.4 35 36 San Rafael Creek 800 120 61 38 M 350 37 Corte Madera Creek 140 12 11 8.7 48 81 38 Cone Madera Creek 66 42 42 6.7 £1 11 39 Arroyo Corte Madera del Persidio up 34 7.6 11 16 24 ' Based on oven-dry weight of stream bed material uncorrected for per- cent recovery. - Underlined values indicate mass spectrometric confirmation of residue identity. ' The presence of 55 «g/kg of polychlorinated naphthalenes (PCN) obscured any PCB's present. PCN was identified by gas chromato- graph/mass spectrometer. bution in the San Francisco Bay area. Initially only the presence of the DDT family of compounds and the PCB's had been anticipated with any certainty, due to their established ubiquity. This investigation soon showed the insecticide chlordane to be as prevalent as the other target compounds; it appeared in 92 percent of the 39 35 samples analyzed. Both the PCB's and chlordane evi- denced a wide range in concentration, from 1 to greater than 1,000 /xg/kg. In spite of this extreme range there does not seem to be a significant difference between the average residue concentrations of streams dischargmg into the Bay south of San Francisco and those discharg- ing into the Bay north of San Francisco. Significantly, only 3 of the 39 streams tested contained residues of the DDT family in quantities greater than those of PCB or chlordane. Although a widespread occurrence of PCB's had been expected, two sampling sites showed residue levels much higher than anticipated. On Stevens Creek site 10 had residue levels of 180 /xg/kg; on Alamitos Creek site 16 had levels of 610 /xg/kg. Neither area has any apparent industrial or commercial development. The sample from site 14 on the Guadalupe River contained 55 /xg/kg of polychlorinated naphthalenes. This is an industrial com- pound similar in properties and uses to PCB's. This sample, too, came from an area of no apparent industrial activity. Nonachlor, a component of commercial-quality chlordane, was often found in relatively higher concen- trations in bottom material samples than in freshly pre- pared standards. This suggests that nonachlor may be more resistant to degradation than either the a or y isomers of chlordane. Acknowleclgincnt The authors wish to acknowledge the contribution of D. H. Peterson, U.S. Department of the Interior — Geo- logical Survey, for providing the samples used in this study. We would also like to thank Miss Victoria A. Wright for her able assistance in the laboratory. LITERATURE CITED (1) Giaiti. C. S.. A. R. Hanks, R. L. Rkliardson. W. M. Sockett. and M. K. Wong. 1972. DDT. DDE. and poly- chlorinated biphenyls in biota from the Gulf of Mex- ico and Caribbean Sea — 1971. Peslic. Monit. J. 6(3): 139-143. (2j Holden, A. V.. and K Maisden. 1967. OrganiKhlorine pesticides in seals and porpoises. Nature 216:1274-1276. (3} Holmes. D. C, J. H. Simmons, and J. O'G. Talten. 1967. Chlorinated hydriKarbons in British wildlife. Na- ture 216:227-29. (4} Karlog. O.. 1. Kiaul. and Sv. Dalgaard-Mikkclscn. 1971. Residues of polychlorinated biphenyls (PCB's) and or- ganochlorine insecticides in liver tissue from terrestrial Danish predatory birds. Acta. Vet. Scand. 12:310-312. (5) Koeman, J. H.. M. C. ten Noever de Braitw, and R. H. de Vos. 1969. Chlorinated biphenyls in fish, mussels and birds from the River Rhine and the Netherlands coastal area. Nature 221:1126-1128. (6) Modin. J. C. 1969. Chlorinated hydrocarbon pesticides in California bays and estuaries. Pestic. Monit. J. 3(l):l-7. (7) Riseb rough, R. W., P. Ricche. D. P. Pcakall, S. G. Herman, and M. N. Kirven. 1968. Polychlorinated biphenyls in the global ecosystem. Nature 220:1098- 1102. (5) Schmidt, T. T.. R. W . Risebroiigh. and F. Grcss. 1971. Input of polychlorinated biphenyls into California coastal waters from urban sewage outfalls. Bull. En- viron. Contam. Toxicol. 6(3):235-243. (9) Sodergren, A.. Bj. Svensson, and S. Ulfstrand. 1972. DDT and PCB in South Swedish streams. Environ. Pollut. 3(0:25-36. (10) Tombergs. H. P. 1972. The PCB situation in Germany. Environ, Health Peispect. 1:179-180. (11) Zitko. V. 1971. Polychlorinated biphenyls and organo- chlorine pesticides in some freshwater and marine fishes. Bull. Environ. Contam. Toxicol. 6(4): 464-470. (12) Sanders. H. O., and J. H. Chandler. 1972. Biological magnification of a polychlorinated biphenyl (Aroclor z S sg Z as DDE Dieldrin Heptachlor epoxide PCB's (polychlori- nated biphenyls) ODD DDT Endrin DDMU. Aldrin 127 58 23 :o 10 3 ■} 1 94.8 .'0.7 17.2 14.9 7.5 2 2 1.5 1.5 0.7 100 69 32 13 13 8 5 0 3 98.0 67.6 31.4 12.7 12.7 7.8 4.9 0.0 2.9 65 19 7 15 1 3 0 0 1 89.0 26.0 9.6 20.5 1.4 4.1 0.0 0.0 1.4 ' Total number of golden eagles represented = 169. Table 3 gives the median, range, and number of sam- ples with each organochlorine insecticide residue for each sample type according to State. These data are generally too variable and sample size is too small to permit meaningful statistical comparisons between States or regions. From the data available, it appears that ex- posure of individual golden eagles to organochlorine insecticides is quite variable but falls within approxi- mately the same range across the country. As shown in Table 3. DDE was found in at least one golden eagle from each of the 22 collection States. Few of the samples contained alarmingly high residues, how- ever. Four of the fat samples contained over 40 ppm DDE (50-84 ppm). two of the BHKLM samples con- tained over 20 ppm DDE (24-25 ppm), and four of the brain samples contained over 2 ppm DDE (2.3-9.9 ppm). According to the general guidelines given by Stickel et al. (4) in their study of DDE residues in cowbirds (Molothrus liter), these residues are not sufficiently high to cause direct, nonsynergistic acute toxicity to eagles. Golden eagles from all 22 States contained a median of 0.3 ppm DDE in BHKLM. 3.0 ppm in fat. and 0.1 ppm in the brain for the years 1964-70. These are higher levels than those of DDT and DDD in all sample types, indicating that most exposure to DDT and metabolites was not recent or was dietary. The DDE levels, however, are considerably lower than the lowest median DDE residue of 4.92 ppm in the carcass and 0.92 ppm in the brain of bald eagles (Haliaeetus leucocephahts) as re- ported by Mulhern et al. (5) for the years 1966-68, and 7.80 ppm in the carcass and 1 .00 ppm in the brain of bald eagles as reported by Reichel et al. (6) for the years 1964-65. The lower residue content in the golden eagle compared to that in the bald eagle probably re- flects their differing diets. The golden eagle's preferred diet of terrestrial herbivores such as rabbits and rodents (7,8.9) places it at the end of a shorter food chain than that of the bald eagle, which frequently takes fish ilO). 38 Pesticides Monitoring Journal Reichel et al. (6) also presented residue values for 21 golden eagle samples collected in 1964 and 1965, mostly in South Dakota. These birds also contained residues lower than those in bald eagles, with median values of 0.49 ppm DDE and 0.09 ppm dieldrin in the carcass, and 0.10 ppm DDE and less than 0.05 ppm dieldrin in the brain. The 40 golden eagle samples from South Da- kota for the years 1964-70 presented in Table 3 con- tained similar residue levels: median values of 0.2 ppm DDE and less than 0.1 ppm dieldrin in BHKLM; less than 0.1 ppm DDE and 0.1 dieldrin in the brain. TABLE 3. — Organochlorine residues in golden eagles, listed by compound and Slale- Residues in ppm, wet-weight basis -1964-70 State BHKLM Brain Fat Median Range Ni Median Range N' Median Range Ni DDE Alaska 9.3 — 1 1.0 — 1 — — — Ariz. — — — — — — 4.1 — 1 Ark. — — — — — — 0.3 — 1 Calif. 0.5 <0.1-5.2 6 0.2 <0.1-2.4 6 0.4 <0.1-I6 5 Colo. 0.3 <0.1-6.5 12 <0.1 — 4 0.8 <0.1-15 10 Idaho 4.0 0.1-8.0 2 <0.1 <0. 1-0.3 3 1.8 0.7-3.0 2 111. 0.1 — 1 — — — 1.6 — 1 Ind. 1.6 — 1 — — — 13 — 1 Iowa <0.1 <0.1-0.1 2 — — — — — — Md. 3.2 0.7-5.7 2 2.3 — 1 84 — 1 Minn. 0.6 < 0.1-7.9 7 0.4 <0. 1-0.7 4 7.1 1.8-40 5 Mo. 0.4 0.3-3.1 3 0.8 0.2-1.4 2 <0.l — 1 Nebr. 0.2 <0. 1-2.6 26 <0.1 <0.1-1.1 U 2.5 < 0.1-26.0 28 Nev. 0.3 — 1 0.1 — 1 — — — N. Mex. 0.1 — 1 — — — 1.6 — 1 N. Dak. 0.3 < O.I -24 8 <0.1 <0. 1-0.2 4 3.4 1.3-50 8 Okla. 0.5 _ 1 — — — 10 — 1 Oreg. — — — 1.2 — 1 28 1.3-55 2 Pa. 1.0 0.4-6.8 4 (1,2 <0.1-0.5 4 29 6.6-38 4 S. Dak. n,2 <0.1-25 41 <0.1 <0. 1-4.6 18 4.4 1 .0-28 23 Utah 0.4 <0.1-4.0 6 0.5 <0.1-1.9 3 3.6 0.3-55 4 Wis. 6.2 1.4-11 2 5.0 0.1-9.9 2 12 — 1 All States 0.3 <0.1-25 127 0.1 < 0.1 -9.9 65 3.0 <0.1-84 100 DIELDRIN Calif. — — — — — — <0.1 — 1 Colo. <0.1 <0. 1-4.4 8 <0.I — 3 0.8 <0.1-12 10 Idaho <0.1 — 1 — — — — — — 111. 0.1 — 1 — — — 1.3 — 1 Ind. 0.2 — 1 — — — — — — Iowa <0.1 — 1 — — — — — — Minn. 0.1 <0. 1-0.2 3 <0.1 — 2 0.7 0.4-1.9 4 Mo. — — — <0.1 — 1 <0.1 — 1 Nebr. <0.1 <0. 1-0.4 15 <0.1 <0. 1-0.1 3 0.4 <0.l-2.2 22 Nev. 0.1 ^ 1 <0.1 — 1 — — — N. Mex. <0.1 — 1 — — _ <0.l — 1 N. Dak. <0.1 <0.1-1.5 4 <0.1 — 1 1.4 0.4-3.1 6 Okla. <0.1 — 1 — — — 0.7 — 1 Pa. 0.2 — 1 — — — 2.4 — 1 S. Dak. <0.l <0. 1-3.0 27 <0.l <0. 1-0.4 8 0.8 0.2-6.6 20 Utah 0.1 — 3 — — — <0.1 — 1 All States <0.1 <0.1-4.4 68 <0.1 <0.I-0.4 19 0.7 <0.1-12 69 HEPTACHLOR EPOXIDE Ark. — — — — — <0.1 — 1 Calif. — — — — — — <0.1 — 1 Colo. Md. Minn. 0.1 < 0.1-0.2 3 <0.1 — 2 0.4 1,4 <0. 1-1.2 g <0.1 1 Mo, 0.2 — 1 0.1 — 1 <0.1 — 1 Nebr. <0.1 <0. 1-0.1 11 O.I — 1 0.7 <0.1-1.2 9 N. Dak. 0.4 — 1 — — — I.l 0.5-2.4 3 S. Dak: 0.2 <0. 1-12.2 6 0.3 <0.1-2.3 3 0.5 0.2-1.3 7 Utah — — — — — — <0.1 — I All States <0.1 <0.1-12.2 23 0.1 <0.I-2.3 7 0.4 <0.1-2.4 32 June 1974 39 TABLE -i.—Oigaiiochlorinc residues in golden eagles, listed by compound and State— 1964-70— Continued Residues in ppm, wet-weight basis State Ark. Calif. Colo. Nebr. N. Mex. Okla. Pa. S. Dak. All States Ark. Colo. Idaho Minn. S. Dak. Utah All States Colo. Md. Nebr. N. Dak. All States Colo. Calif. Colo. S. Dak. All States BHKLM Range Ni Brain Median Median DDD 0.2 <0.1 <0.1 ..0.1 n.i 0.2 1.6 <0.1-2.4 5 1967 <0 I <0.1-2.3 24 ■.'0.1 _ 1 0.5 <0.1-3.1 24 0.1 <0. 1-0.6 17 <0.1 <0.1-0.4 9 1.2 <0. 1-6.6 17 1969 1970 •^O.I <0.1-0.4 12 <0.1 — 7 0.7 0.2-4.6 10 1964-70 <0.1 < 0.1-4.4 68 <0.1 <0.1-fl.4 19 0.7 ^,0.1-12 69 H EPTACHLOR EPOXIDE 1964 — — — _ 0.4 <0.1-1.2 8 1965 <0 1 0.8 0.5-1.1 2 1966 <0.1 <0. 1-0.2 4 — — — 1.2 <0.1-2.4 2 <0.1 <0.1-12.2 10 <0.1 — 1 0.7 <0.1-1.4 13 0.1 < 0.1-0.6 4 <0.1 <0.1-2.3 4 0.4 <0. 1-1.2 4 1970 <0.1 <0.1-0.2 4 0.1 — 2 0.2 <0. 1-0.4 3 1964-70 <0 1 <0.1-12.2 23 0.1 <0.!-2.3 7 0.4 <0.1-2.4 32 DDD 1964 — <0.I 4 1965 8.0 _ 1966 <0.I <0. 1-0.3 — _ 1.2 0.8-1.6 2 1967 <0.1 — — 0.2 <0. 1-1.0 4 1968 0.1 — _ _ 2.0 1 1969 0.1 <0.1-0.2 2 <0.1 I I.O 0.9-1.2 2 1970 — 1964-70 <0.1 <0. 1-8.0 10 <0.1 - 1 0.5 < 0.1 -2.0 1 1964 1965 — — — - — — <0.1 - 6 1966 1967 1968 0.2 <0.1 2 I <0.1 — 1 <0.1 <0.1 - I 1 1969 1970 — — — 0.1 < 0.1 -0.2 2 - - - 1964-70 0.2 < 0.1-0.2 3 <0.1 <0.1-0.2 3 <0.1 — 8 41 TABLE 4. — Organochlorine residues in golden eagles by year — 1964-70 — Continued Residues in ppm, wet-weight basis Year BHKLM Median Range Median N' Fat Median Range ENDRIN 1964 — — — — 1965 <0.1 — 1 — — — <0.1 — 1 1966 <0.1 — 1 — — — — — — 1967 1968 1969 1970 1964-70 — — <0.1 <0.1-0.3 4 <0.l - 2 - - - <0.1 <0. 1-0.3 5 DDMU 1964 I96S 1966 1967 0.6 0.5-0.6 2 - - - - - - 1968 1969 1970 1964-70 0.6 0.5-0.6 2 - - - - - - ALDRIN 1964 — _ — — — — <0.I <0. 1-0.1 3 1965 1966 1967 1968 1969 1970 1964-70 — — — <0.1 — 1 — — — <0.1 — ' — — - - - - <0.1 — 1 <0.l — - <0.1 <0. 1-0.1 3 ' Number of specimens containing residue. Median is based on this number. pies (for median levels in birds from all States, see Table 3). Each of these insecticides had a median of less than 0.1 ppm in the brain. The data in Table 4 suggest that exposure of golden eagles to organochlorine insecticides has not changed significantly from 1964 through 1970. The years with the largest sample sizes (1966-69) indicate no important changes in exposure. The median concentrations of DDE, which is represented by the largest number of samples, ranged only from 0.2 to 0.4 ppm in BHKLM samples, and from 2.8 to 4.2 ppm in the fat samples during these years. This is similar to the findings re- ported by Mulhern et al. (5) for bald eagles. Cases in which necropsy or residue analysis indicated the probable cause of death are listed in Table 5. Of TABLE 5. — Probable cause of dealli of 169 golden eagles as determined by necropsy or residue analyses Probable cause or death Number Shooting 18 Impact on power line or electrocution 15 1080 (sodium monofluoroacetate) 11 Trapping 10 Collision with vehicle 5 Strychnine 2 Cyanide 2 Unknown 106 the 63 eagles included. 33 died from shooting or contact with power lines (collision or electrocution). Six of the eagles killed by 1080 (sodium monofluoroacetate) were collected in 1966. three in 1968, and two in 1969. Most of the specimens examined died from undetermined causes, which undoubtedly included a high proportion of natural deaths. LITERATURE CITED (1) Johnson. R. E.. T. C. Carver, and E. H. Dustman. 1967. Residues in fish, wildlife, and estuaries. Pestic. Monit. J. Ul):7-I3. (2) Mills, P. A. 1959. Detection and semiquantitative esti- mation of chlorinated organic pesticide residues in foods by paper chromatography. I. Assoc. Off. Agric. Chem. 42:734-740. (3) Storherr. R. W., and R. R. Watts. 1965. A sweep co- distillalion cleanup method for organophosphate pes- ticides. I. Recoveries from fortified crops. J. Assoc. Off. Agric. Chem. 48(6): 1154-1158. (4) Slickel, W. H., L. F. Stickel, and F. B. Coon. 1970. DDE and DDD residues correlated with mortality of experimental birds. Inter-Amer. Conf. Toxicol. Occup. Med. pp. 287-294. (5) Mulhern, B. M., W. L. Reichel, L. N. Locke, T. G. Lamont, A. Belisle, E. Cromartie, G. E. Bagley, and R. M. Prouty. 1970. Organochlorine residues and au- topsy data from bald eagles. Pestic. Monit. J. 4(3): 141-144. 42 Pesticides Monitoring Journal (6) Reichel, W. L., E. Cromartie. T. G. Lanumt, B. M. Mulhent, and R. M. Prouly. 1969. Pesticide residues in eagles. Pestic. Monit. J. 3(3): 142-144. (7) Boekei; E. L., and T. D. Ray. 1971. Golden eagle popu- lation studies in the Southwest. Condor 73(4):463-467. (8) McGahan, J. 1968. Ecology of the golden eagle Auk 85(0:1-12. (9) McGahan. J. 1967. Quantified estimates of predation by a golden eagle population. J. Wildl. Manage. 31(3)- 496-501. {10} Imler, Ralph //.. and E. R. Kalmbach. 1955. The bald eagle and its economic status. U.S. Fish WiidI Serv Circ. 30. (//) Slickel, W. H., L. F. Stickcl, and J. W. Spann. 1969. Chemical fallout; current research on persistent pesti- cides. Proc. First. Rochester Conf. Toxicity, pp. 174- 204. (12) Lockie. J. D.. D. A. Ralcliffc. and R. Balharry. 1969. Breeding success and organochlorine residues in golden eagles in West Scotland. J. Appl. Ecol. 6(3):38 1-389. (13) Hickey, J. J., and D. W. Anderson. 1968. Chlorinated hydrocarbons and eggshell changes in raptorial and fish-eating birds. Science 162(3850):271-273. (14) Ralcliffe, D. A. 1970. Changes attributable to pesticides in egg breakage frequency and eggshell thickness in some British birds. J. Appl. Ecol. 7(1):67-107. Vol. S, No. 1, June 1974 43 Toxaphene Content of Estuarine Fauna and Flora Before, During, and After Dredging Toxaphene-Contaminated Sediments' Robert J. Reimold and Charles J. Durant ABSTRACT This paper evaluates loxaphenc concentrations in selected estuarine fauna, flora, sediment, and dredge spoil before, during, and after the dredging of Teiry Creek. Brun.swick. Ga., in autumn 1972. This is the second effort to widen the channel of the creek, which receives the effluent from a nearbv toxapliene-manufacturing plant: a 1971 dredging ef- fort was aborted by the Stale of Georgia. The current study employed safeguards inspired by the 1971 State action: en- closure of dredge spoil in diked areas of unproductive marsh- land to prevent runoff, and weekly monitoring of Terry Creek biota and sediment to detect the possible role of toxaphene in any resulting disturbance to the balance of nature. Moni- toring of dredge spoil, fauna, and flora showed toxaphene concentrations to be higher during dredging than before or after. Eastern oysters (Crassostrea virginica). reported to be among tlie best biological monitors, did not demonstrate large changes in toxaphene content resulting from the dredg- ing. The high toxaphene concentration in oysters ranged be- tween 2.0 ami 5.0 ppm. The best indicators evaluated were salt marsh cordgra.fs (Spartina alterniflora) and mummichog (Endulus heteroclitiis). The highest content found in S. alter- niflora was 7.5 ppm: the highest concentration in F. hetero- clitiis was over 200 ppm. Introduction A unique occasion to study the dispersal of the chlori- nated hydrocarbon, toxaphene, and the relation of its concentration to waterway maintenance dredging arose near Brunswick, Ga. Terry Creek, which receives the effluent from a toxaphene-manutacturing plant, was the Contribution No. 30. University of Georgia Marine Institute. Sapclo Island, Ga. 31327. site of a recent dredging operation. An earlier operation (/) had been aborted after 100 yards of dredging, when the State of Georgia objected that toxic products in the undiked spoil sediment might drain through the marsh- land on which the spoil was being deposited. Such drainage would contaminate both the potentially pro- ductive marshland and the estuarine ecosystem into which it drained. The current study initiated safeguards against such a contingency: enclosure of dredge spoil in diked areas of marshland which had been identified by remote sensing techniques C3) as less productive than the marshland initally used as deposit sites; and weekly monitoring of Terry Creek biota and sediment to detect the possible role of toxaphene in any resulting disturb- ance to the balance of nature. The monitoring program which the U.S. Army Corps of Engineers, Savannah District, outlined in response to the State of Georgia was designed to document at weekly intervals the possible role of toxaphene in any ecologic disturbance resulting from the dredging of Terry Creek. Toxaphene concentrations were measured in selected estuarine fauna, flora, sediment, and dredge spoil during dredging. Findings were compared to the authors' earlier baseline studies of pesticide concentra- tions in the estuarine ecosystem, which revealed toxa- phene in many portions of the food web and highly concentrated in the sediments of the creek (4). Methods Terry Creek was dredged by hydraulic pumping anc 5 X 10'' cubic yards of spoil was placed in two dikec 44 Pesticides Monitoring Journai areas on a nearby salt marsh (areas C and D, Fig. 1). Collection sites for environmental samples are indicated in the same figure: A) Back River Bridge; B) mouth of Terry Creek; C) west dredge spoil area near the toxa- phene plant outfall; D) east dredge spoil area near Back River; and E) main channel of Terry Creek. Field col- lections were obtained at weekly intervals beginning one week before dredging operations started on September 7, 1972, and terminating 8 weeks later, 1 week after the completion of all dredging on October 26, 1972. Addi- tional baseline data on the toxaphene content of Terry Creek organisms were collected during 1970 and 1971 and followup collections continued at monthly intervals for several months. Environmental samples included finfish collected by otter trawl and cast net in the main channel of Terry Creek; salt marsh cordgrass (Spartina aherniflora) collected near the dredge spoil dike areas; sediment collected from the mouth of Terry Creek and from within each of the diked spoil areas; and oysters {Crassostrea virf^inica) collected from Back River Bridge. FIGURE 1. — Geographic location of collection .'.iles near Terry Creek, Brunswick. Ga.. 1972 Samples were processed and analyzed for toxaphene ac- cording to authors' methods in the earlier baseline studies [4) and the technique of Wilson (5). Specific de- tails are summarized in Table I. Chromatograms of environmental samples were so similar to chromato- grams of technical grade toxaphene standard that no polychlorinated biphenyls were indicated. All concen- trations are expressed in parts per million (ppm) wet weight except S. aherniflora which is based on dry weight. Prior to analysis, 5. alternifiora plant parts were given four rinses in tap water to remove surface salt deposits. Each leaf and stem was then blotted dry with tissue to remove residual rinse water. The relative re- covery from oysters and sediment was above 85 percent and 90 percent, respectively. Data were not corrected for this error and concentrations below 0.25 ppm were considered insignificant with the exception of those in- cluded in water samples. Concentrations less than 0.0010 ppm were considered below the limits of detec- tion. TABLE I. — Summary of column packing and operating parameters used for toxaphene analysis Colu 5 ft b.v '/'« in., glass, packed with 3% DC-200 on Gas Chrom Q. 80/100 mesh Temperalure: Dcleclor 210°C Injector 210°C Oven I90°C Carrier: Prepurified nitrogen at a flow rate of 40 ml/min Results Results of toxaphene analyses in fauna, flora, and sedi- ment reveal that toxaphene was generally higher in dredge spoil sediment than in any other samples proc- essed (Table 2). Comparisons of concentrations in mum- michog (Fundulus heteroclitus). white shrimp (Penaeus setiferus), marsh grass, and sediment show that, except for dredge spoil within the diked areas, concentration rarely exceeded 10.0 ppm (Fig. 2-10). Butler (6) demonstrated that shellfish can produce bio- logical magnification of pesticide levels up to 70,000 times those of surrounding water. Yet the levels in oys- ters collected at Back River Bridge (Fig. 1 ) adjacent to the mouth of Terry Creek never exceeded 2.0 ppm dur- ing the dredging operation. Toxaphene concentrations in dredge spoil from the wes- tern enclosure close to the toxaphene plant outfall (Fig. 1) neared 1,000 ppm (Fig. 5). The highly contaminated sediment in both the western and eastern enclosures was held within two dikes (Fig. 1) and did not appear to influence the surrounding biota. Background toxaphene concentrations in 1970-71 col- lections of fauna, flora, and sediment exhibited higher values than 1971-72 concentrations. The latter values remained about the same before and after dredging, reflecting in part the pollution abatement practices initi- ated at the toxaphene production plant which greatly decreased the quantities of toxaphene in the plant effluent. Discussion Toxaphene contamination appears to be best indicated by the marsh grass {S. aherniflora). Various species of fish are also good indicators of the increased toxaphene in the suspended material and water. Although Butler (6) has suggested that shellfish are among the best bio- logical monitors for pesticide residues, the results of this study suggest that marsh grass and finfish accumulate toxaphene to a greater degree than oysters (Crassostrea virginica) when all are collected from the same geo- VoL. S, No, 1, June 1974 45 TABLE 2.—Toxaphene concentrations, ppm. found during jail 1972 monitoring oj Terry Creek dredging operation' Mummicliog (Fundiitus lieleroclitus) Salt marsh cordgrass {Sparlina allerniflora) Eastern oyster (Crasioslrea virginica) Sediment (near entrance to Terry Creek) Dredge spoil from east enclosure ( Back R. ) Dredge spoil from west enclosure (Dupree C.) Water sample Anchovy (Anclioa mitcl^elli) Shrimp {Penaeus setijerus) head and thorax Shrimp ( Penaeus seliferus ) abdomen (edible tail) Star drum IStelltfct taiHeolatus) Repli- cate No. Cruise No. 1 (Sept. 2) 10.45 8.97 0.82 0.76 1.20 1.37 5.47 4.42 NSC NSC NSC NSC NO ND NSC NSC NSC NSC NSC NSC NSC NSC Cruise No. 2 (Sept. 14) NSC NSC 1.13 1.56 1.42 1.33 5.56 4.20 0.81 0.79 NSC NSC NSC NSC 8.61 NAS 1.21 1.51 0.58 0.58 2.43 2.61 NSC— no sample collected; NAS— not adequate sample; ND— not detectable. IDnuhk l!jn liulHJW Kl'pIuiiic MiiiiU-. ,ulh;h'J /•>7l).7l 71-7: 7U-7I ' mcjn ul /v 70 71 1 71 7: WtEKLY CRUISt NO Cruise No. 3 (Sept. 21) Cruise No. 4 (Sept. 28 NSC NSC 0.73 0.56 1.55 1.71 0.87 0.94 60.08 150.79 NSC NSC 0.0013 NAS NSC NSC 4.80 4.65 0.86 0.90 2.09 3.24 10.52 3.41 2.04 2,54 1.21 1.15 2.11 2.68 5.70 5,12 NSC NSC 0.0013 NAS 20.46 16.60 3.22 2.24 1.22 0.88 NSC NSC Cruise No. 5 (Oct. 5) NSC NSC 0.81 0.91 1.26 1.23 5.55 7.24 NSC NSC 93.39 119.52 0.0016 NAS 8.96 NAS 3.03 2.85 0.92 0.73 2 72 2.48 Cruise No. 6 (Oct. 12) 131.14 217.14 3.93 3.68 1.19 1.24 3.97 4.06 NSC NSC 756.40 812.54 0,0014 NAS 9.89 10.13 NAS 1.49 NAS NSC NSC Cruise No. 7 (Oct. 19) 12.07 5.18 7.33 6.26 1.70 1.79 2.11 2.23 NSC NSC 51.64 142.42 NSC NSC 12.85 11.96 4.19 4.30 0.74 0.64 Cruise No. 8 (Oct. 26) NSC NSC NSC NSC 1.69 2.92 0.94 0.95 2.54 3.74 NSC NSC 241.54 331.55 NSC NSC 10.51 NAS 10.69 2.41 2.51 0.83 1.42 1,09 FIGURE 2. — Toxaphene concentration, ppm. in oysters (Crassostrea virginica) from Back River Bridge, Terry Creek. Brunswick, Ga., 1972 FIGURE 3. — Toxaphene concentration, ppm. in surface sedi- ment from trtouth of Terry Creek, Brunswick, Ga., 1972 46 Pesticides Monitoring Journal FIGURE 4. — Toxaphene concentration, ppin, in dredge spoil from east enclosure near Terry Creek. Brunswick. Ga.. 1972 graphic location. The authors submit that different organisms may be able to preferentially concentrate different residues. Although relatively high concentra- tions of toxaphene were found in anchovy (AnclxKi mitchelli). it was frequently absent from the trawl catch. The high toxaphene content of .S. alierniflora is of in- terest since it represents a translocation of toxaphene from the sediments into the plant tissue. S. alierniflora is a halophyte and probably translocates toxaphene along a pathway similar to the movement of salt. This is unique because it is one of the only known instances of toxaphene translocation in high concentrations among primary producers. Altogether, over 125 samples were analyzed for toxa- phene during the 8-week study. In addition, qualitative visual examination of the Terry Creek area was made at weekly intervals to assess potential fish kill. Fewer than 20 dead fish were recovered during the 8 collection trips of 4 to 5 hours each. Those recovered and posi- tively identified included tongue-fish (Symphurus sp.) harvest fish (Peprihts sp.). silver perch (Bairdiella sp.). sea catfish (Arius sp.), and one menhaden (Brevoortia sp.). IDtiiihlc Bur, hiduiiw Rc/itnaU' SainpU,} WH-KLV CRUlSbNU J FIGURE 5. — To.xaphene concentration, ppiii. in dredge spoil from west enclo.mre near Terry Creek. Brunswick. Ga.. 1972 Local shrimp bouts that dock in Terry Creek frequently offloaded and sorted catches during the study; this may well have been the source of dead fish, especially since the authors" sampling indicated that the species were not indigenous to Terry Creek. At no time were fish observed to be dying (alive but floating on the surface on their sides), nor did they show other signs of distress during the nearly 100 person-hours of field sampling in Terry Creek. It is the conclusion of this study that dredging did not dramatically alter the biological balance of the estuary by toxaphene contamination when toxaphene residues were isolated in diked enclosures. As reported by the authors in the baseline studies (4), the combination of ultraviolet radiation and biological degradation should render the impounded sediments nontoxic in a few years. Vol. 8, No. 1, June 1974 47 SEED HEADS X = 4,93 N= 2 S = 1.92 N = Number of Observations X = Mean Toxaphene Con- centration S = Standard Deviation LEAVES X= 36.30 N= 10 S = 4.51 SEDIMENT X= 32.55 N = 4 S = 22.15 ROOTS X= 1.91 N = 1 RHIZOMES X = 1.24 N= 1 FIGURE 6. — Toxaphene concentration, ppni, in maisli grass (Spartina alterniflora) and surrounding sediments collected from Terry Creek Marsh. Brunswick, Ga., 1970-72 ..iii,,il. , . ..II.. I,.l li''il-'l 'I ' I'fl--: I i ..iiitrl. • . -11. > '' J rfl I ki ^ ( KLIM M' FIGURE 8. — Toxaphene concentration, ppm, in white shrimp (Penaeus setiferus) from Terry Creek. Brunswick, Ga., 1972 FIGURE 7. — Toxaphene concentration, ppm, in marsh grass (Spartina alterniflora) leaves anil stems from bank of Back River near Terrv Creek, Bnin.'iwick, Ga., 1972 li)."il>U- Hjf, hiJu-iU K.iiliijW ij(»l/>/<' 711-71 = „r,j„ .,/ v ■ .iinpl.M'ilU-.i.d fJ'ii-'l 71-7: - >ii,.i,i ..I II ,.wti'k^,.'lli'.t,uplicate samples were placed in sealed quart jars in a 'OL. 8, No. 1. June 1974 dry ice bin for periods of one hour, and for one, three, and seven days. The pH of the water varied from 6.7 to 6.9. These samples were washed three times with ethyl acetate in the same manner as the plant material. Since resmethrin is insoluble in water, no difficulties were encountered with the extraction procedure using 500 fil of ethyl acetate. The ethyl acetate was evaporated to a volume of 100 jxl of which 10 fxl was spotted on a TLC plate. The field samples of water were taken from the lower, middle, and upper parts of a stream in each plot. Forty grams of each water sample was extracted three times with 500 ml of ethyl acetate. The samples were ana- lyzed in duplicate in the same manner as the water con- trols. The small amount of debris in the field stream samples presented no problems during extraction. The pH varied from 5.4 to 5.8. Results and Discussion Immediately after the resmethrin was sprayed, no de- tectable residues were found on the aspen at the lower dose rates, but at the 0.15-lb rate, 0.8 ppm was found (Table 1). Only the willow samples showed a deposit as high as 1 ppm. The initial deposit was light; after seven days, no detectable residues were found. The <0.05 ppm designation was used when very faint traces were visible on the TLC. The results suggest that after aerial application of resmethrin at 0.05 and 0.15 lb/ acre, no substantial residue remains after seven days. TABLE I. — Resiuelhrin residues, ppm, found days after aerial application on tliree types of plant foliage Plant Days after application Control forti- fied WITH 0.5 PPM RESMETHRIN ' RATE: 0.05 LB/ ACRE Aspen 0 0 0 0 0 0 0 0 0 0.4 Wiliow 1.0 0.3 <0.05 1.0 0.3 0 1.0 0.3 <0.05 0.5 Douglas hr 0,3 0 0 0.3 0 0 0.8 0 0 0.4 RATE: 0.15 LB/ACRE Aspen 0.8 0 0 0 0 0 0.4 0 0 0.5 Willow 0.8 0,5 <0.05 0.5 0 0 0,5 0.4 <0.05 0.5 Douglas iir 0.3 0 0 0.5 0 0 0.5 0 0 0.4 Average of six samples analyzed with an accuracy of ±7%. 51 Recovery of the 10 and 20 jig of the fortified water controls with resmethrin on the TLC plates were meas- ured and remained unchanged within a 10 percent error. The water samples from the field showed no detectable quantities of resmethrin. Although the lesser application rate of 0.05 lb/ acre of resmethrin had a fatal effect on most of the aquatic insects in the 1-hour postspray period, no toxic effects were observed in fish held in cages in the stream (5). The fact that no res- methrin was recovered from the aquatic samples even though insect mortality was high obviously indicated substantial degradation, considerable dilution of the insecticide, or insufficient understanding of resmethrin within the environment. LITERATURE CITED (/) Unpublished repoil. March I. 1971. Pacific Southwest Forest and Range Experiment Station, Berkeley, Calif. (2) Elliot, M.. A. W. Fainham, N. F. James, P. Needham, and B. C. Pearson. 1967. A new potent insecticide. Na- ture 213, (5075) 493-494. (3) Curtis. P. F., and G. T. Phillip. 1962. Isatin as a chro- mogenic agent for detection of thiopene. J. Chromatogr. 9(11): 366". (4) Abernathy, C. O., J. E. Casida, and I. Yamamoto. 1971. Division of Entomology, University of California, Berke- ley, Calif. Unpublished resuhs. (5) Shea, P. J. 1971. Unpublished report. Pacific Southwest Forest and Range Experiment Station, Berkeley, Calif. 52 Pesticides Monitoring Journai Detection of DC PA Residues in Environmental Samples F. M. Miller and E. D. Gomes ABSTRACT DC PA was detected in river water, five species of fish, and air in the lower Rio Grande Valley of Texas. Its presence was determined by electron-capture gas-liquid chromatog- raphy and confirmed by several analytical methods. Analyses of water samples taken over a 2-year period usually indicated less than I ppb DCPA in water during both years. Residues in one freshwater and four saltwater fish species varied from less than I ppb to 8 ppm. DCPA was found in air samples for several months following use in a vegetable- growing area. Introduction DCPA (Dimethyl tetrachloroterephthalate) is a herbicide that is sold under several brand names for control of grasses and weeds in turf and gardens and for use in commercial agriculture. In the lower Rio Grande Valley, DCPA is used primarily as a pre-emergence herbicide on onion soils and to a lesser extent on cabbage and cotton crops. This chlorinated hydrocarbon pesticide is of low solu- bility, is adsorbed by organic matter in soil, and does not leach in any of the general soil types. Its average half-life is 100 days in most soil types and it is relatively nontoxic to animal species that have been tested (/). During 1969 onion soils were sampled in 10 states: 46.5 percent had traces (0.94 ppm) of DCPA. This is in con- trast to a nationwide sampling of croplands during 1969 in which only 0.1 percent of the 1,729 samples tested showed traces of DCPA {2.3). The herbicide has also been detected in air samples from an onion-growing area in Colorado {4). 1 Texas Community Studies— Texas State Department of Health 152 E. Stenger, San Benito. Tex. 78586. This report, which presents data from several studies, indicates rather widespread environmental contamination by a chlorinated pesticide. Aquatic sampling of the two major streams in this area was initiated in 1971 to de- tect differences in pesticide contamination over a period of time. In 1972 selected tissues from two large sam- ples of speckled trout were analyzed to determine the level of chlorinated hydrocarbon pesticides present. Air samples were analyzed for pesticides during 1972 as a part of a national monitoring program (5). Several miscellaneous samples were taken after analytical con- firmation of DCPA. Because contamination of streams and fish has not been reported previously, all available data are presented in this paper. All studies were con- ducted in the lower Rio Grande Valley of Texas, an intensely agricultural area. Methods and Materials .SAMPLING Two sampling sites, one on each of the two major water- ways in the lower Rio Grande Valley, were monitored in 1971 for chlorinated hydrocarbon pesticides. One sample of unfiltered water, bottom sediment, and men- haden (Brevoortia sp.) was taken each month from the Arroyo Colorado at a brackish water location near the port which is east of Hariingen (Fig. I). Only water and bottom sediment samples were taken at the second Rio Grande site near Los Indios. Bottom sediment and water samples were taken in solvent-rinsed containers near the bank; the menhaden were collected by cast netting near the middle of the stream. In 1972 two large samples of spotted seatrout (Cyno- scion nehulosus) were taken by bait casting in the Three Vol. 8, No. 1, June 1974 53 FIGURE 1. Map of Lower Rio G ramie saiiipliiif; sites for DCPA \' alley showiiit; Islands area of the lower Lagiina Madre. One sample was taken during May and the other was collected in July. The length, weight, and sex of each fish were recorded. Each speckled trout was dissected and the ovaries or testes, liver, and 10 g of edible flesh were removed and frozen until analyzed. Several Rio Grande perch (CicMasoma cyanogitttatum) were collected at va- rious freshwater locations along the Arroyo Colorado (Table 1, Fig. 1), and were dissected in the same manner to obtain similar tissues when possible. Collectors caught one mullet {Mui;il sp.) while gathering menhaden. Two speckled trout and one red drum (Sciaenops ocellata) were collected and analyzed for residues of DCPA dur- ing 1973. Air samples were taken from two locations in each ot two cities from February 1972 until January 1973. One MisCo high-volume sampler was placed near the center and one on the periphery of each city. Approximately 61 cubic meters of air were sampled at each location over a 24-hour period. The combined samples from each city were analyzed as one sample. Weekly samples were taken alternately from each city. ANALYTICAL PROCEDURES A 1 -liter water sample was extracted with 150 ml ot hexane in a 2-liter separatory funnel. The contents were shaken vigorously for 2 minutes with venting and al- lowed to settle until the layers separated. The hexane layer was removed and another 15()-ml portion of hexane was added to the water sample. Then the procedure was repeated. The two 150-ml extracts were combined and a small amount of sodium sulfate was added to absorb any water remaining in the extract. The sample was transferred to a 500-ml Kuderna-Danish evaporative concentrator and concentrated to 5 ml followed by Florisil cleanup (6). Each 25-g bottom sediment sample was placed in a 250-ml Erlenmeyer flask, 150 ml of acetonitrile was added, and the sample was shaken for I hour at 3 54 agitations per second on a wrist-action shaker. The en- tire sample was filtered through a solvent-rinsed glass wool filter into a 1 -liter separatory funnel containing 500 ml of water. The soil remaining on the filter was rinsed twice with 25 ml of acetonitrile. The acetonitrile/ water mixture wa.s partitioned twice with petroleum ether. The petroleum/ether extract was concentrated and passed through Florisil for cleanup. Fish samples were analyzed by taking approximately 25 g of menhaden, 10 g of trout flesh, 5 g of perch flesh, and the entire ovaries, testes, or liver. Each sample was weighed and placed in a stainless steel Omni-mixer cup, 50 ml of acetonitrile were added, and the sample was blended at medium speed for 5 minutes. The ovaries were ground in a mortar with sodium sulfate and petro- leum ether. The ground or blended extract was filtered through a solvent-rinsed, glass wool filter into a 1 -liter separatory funnel. The sample filtercake was rinsed with 25 ml of acetonitrile; to this was added 130 ml of petro- leum ether and the mixture was shaken for 2 minutes. Then 600 ml of water was added, the sample was shaken again for about 1 minute, the 2 layers were allowed to separate, and the aqueous layer was discarded. Approxi- mately 200 ml of water was added again. The sample was shaken; the layers were allowed to separate; and the water was discarded again, removing the remaining acetonitrile. The petroleum ether extract was concen- trated and cleaned on a Florisil column. Air samples were collected by drawing atmospheric air through ethylene glycol. The samples were extracted with hexane and were cleaned on a Florisil column (7). When Florisil cleanup was employed, DCPA eluted in the 15 percent fraction. This fraction was concentrated to 5 ml and analyzed by gas-liquid chromatography (GLC). Routine pesticide analyses were accomplished with Micro-Tek 220 gas chromatographs that were equipped as follows: Detector: electron-capture tritium foil; 20 volts. Columns: 1.5% OV-17/1.95% QF-1 on Chromosorb W 100/120 mesh H.P.; 4% SE-30/6'~r QF-1 on Chromosorb W, H.P.; 1.6""^ OV-210/6.4% OV-17 on Gas Chrom Q; 5^; OV-2I0 on Gas Chrom Q. Temperatures: Inlet Detector Columns 225°C 205X 0V-17/QF-1, SE-30/QF-1, and OV-210/OV-17 at 200°C OV-210 at 175°C. Carrier gas: Nitrogen. Flow rate: OV-n/QF-1, 80 cc/min; OV-17/OV-210, 65 cc/min; SE-30/QF-1.75cc'min; OV-210, 60 cc/min. Sensitivity: 1 2 FSD with 50 pg of aldrin. Electrometer setting: 10x16. Pesticides Monitoring Journal TABLE \.—DCPA Residues in Three Speeies of Fisli Date Site of Collected Collection Species Tissue DCPA Residue, ppb November 1972 Arroyo Colorado, Mullet skin 555 Harlingen flesh viscera 159 231 December 1972 Arroyo Colorado, Rio Grande Perch liver 468 Harlingen ovaries flesh 420 217 Kio Grande Perch liver ovaries flesh 388 212 89 February 1973 Arroyo Colorado, Rio Grande Perch liver 215 La Feria ovaries flesh 107 29 February 1973 Arroyo Colorado, Rio Grande Perch liver 190 Mercedes flesh 0 March 1973 Arroyo Colorado, Mercedes Rio Grande Perch ( 1 sample: 2 small fish) flesh 90 March 1973 Three Islands, Kedfish liver 18 Laguna Madrc testes flesh 132 0 Recovery studies were conducted with unfiltered arroyo water and trout flesh. Mean DCPA recovery from water samples was 97 percent, with 88 percent recovery from fish flesh. All results are presented as unadjusted values. CONFIRMATORY TECHNIQUES In addition to GLC analyses, three other confirmatory techniques were used. The extracts of several arroyo water samples (20 gallons) were pooled and the presence of DCPA was confirmed by infrared analysis. The KBr micropellet was scanned on a Perkin-Elmer 337 Grating I Infrared Spectrophotometer. Several fish and water samples were analyzed by a thin- layer chromatography/GLC technique (TLC-GLC) (7). Since there was not enough DCPA to develop on the TLC plate, the Rf value was determined. A portion of the AI;,Om on the TLC plate was removed at the indi- cated point, extracted with hexane, and analyzed on the gas chromatograph Two samples were analyzed by GLC/ mass spectrometry (GLC-MS) ((S). The first sample, a pooled extract of menhaden, was cleaned up by thin-layer chromatogra- phy. When the fish extract showed many interfering substances, a second and much cleaner sample was ob- tained by extracting 10 gallons of arroyo water. Results and Discussion Since 1971 routine samples of soil, water, and menhaden have been taken from the Arroyo Colorado and analyzed by GLC. Some of the unfiltered water samples collected in 1971 showed a very prominent peak which eluted at the same retention time as that of heptachlor epoxide on 0V-17/QF-I and SE-30/QF-1 columns. Analyses of samples in the past have indicated little heptachlor or Vol, 8, No. 1, June 1974 heptachlor epoxide in the Rio Grande Valley. A closer look at this mystery peak revealed a difference of 0,05 relative retention time units from that of heptachlor epoxide on the OV-210/SE-30 column. At that time, the peak was tentatively identified as DCPA. The 1972 water samples showed greater concentrations of the chemical during the fall of the year. Air samples also revealed a substance with an identical retention time. During this same time period, some of the menhaden and trout being analyzed were demonstrating this peak in tissue, liver, and reproductive organs, indicating some persistence of the chemical in the aquatic and estuarine environment. Confirmatory procedures were initiated during the latter months of 1972. CONFIRMATORY PROCEDURES In 1972, the OV-210 GLC column proved to be of great importance in identifying DCPA. The most critical dif- ferentiation was between heptachlor epoxide and DCPA, which elute with almost the same relative retention time on columns other than the OV-210. Most of the com- monly used chlorinated hydrocarbon pesticides elute in the same general sequence when using columns such as the OV-2I0/OV-17 (Fig. 2). With the OV-210 column, however, the relative retention times of some chlori- nated hydrocarbons differ. Heptachlor epoxide increases slightly, but the retention time of DCPA is much greater than that of heptachlor epoxide (Fig. 2). Several samples were analyzed by flame photometric detection, indicating that the substance was not a phos- phorus- or sulfur-bearing compound. The TLC-GLC technique confirmed DCPA in several fish and water samples. A relatively pure sample of DCPA was difficult to obtain, but its presence in arroyo water was confirmed by infra- 55 red spectrophotometry (Fig. 3). The subnanogram amounts of DCPA in arroyo water necessitated the ex- traction of 20 gallons of the water in order to collect enough DCPA for infrared analysis. GLC-MS analyses confirmed the presence of DCPA in water and tish samples. WATER AND SEDIMENT In 1971 DCPA was identified in unfiltered water sam- ples by GLC (Table 2). Trace amounts were found only in water, and were not confirmed by other tech- niques. Similar low residue levels were found through- out 1972 and 1973. Although the monthly sample in September 1972 was negative, an additional sample taken on the same date at a nearby location revealed the highest recorded water value of 132 ppb. Another unfiltered sample taken at the Harlingen site later in September contained 0.91 ppb DCPA. Likewise, two additional samples taken during another negative month, December 1972, contained 0.35 and 0.40 ppb DCPA. It is obvious that monthly sampling does not give an OV-2)0/OV-17 HEPTACHLOR EPOXIDE + DCPA ALDRIN 12 O MINUTES FIGURE 2. — Gas ciiionialogidms showing differences in elulion pattern between OV-2I0 and OV-2W/OV-17 column materials FIGURE 3. — Infrared spectral comparison of DCPA stand- ard and water sample, showing confirmation of DCPA in arrovo water accurate estimate of the amount of DCPA in the stream. Only one DCPA residue of 0.08 ppb was found in water samples from the Rio Grande, possibly because most of the lower Rio Grande Valley drains into the Arroyo Colorado rather than into the Rio Grande. Only one bottom-sediment sample contained a trace (42 ppb) of DCPA. Water samples were analyzed with- out removing the particulate matter; it is not known whether the DCPA residues are in the water or in asso- ciation with suspended particles. MENHADEN Monthly samples of menhaden did not reveal any DCPA residues until March 1972 (Table 2). Each sample in- cluded 25 g of the small fish with the heads and tails removed. Since specific tissues were not analyzed, these results cannot be directly related to the values for other fish. Based on current procedures, however, it appears that the menhaden contain greater amounts of DCPA than the other species of fish sampled to date. The higher residue values probably are not due to contami- nation from the gut contents because the gut was ex- tracted separately on several occasions and residues were less than those in the remainder of the tissues. As with the water residue, the amount of DCPA in men- haden varied widely. In November an additional sample taken at a site nearby contained 7.27 ppb DCPA; the regularly scheduled sample had only 0.52 ppb. OTHER FISH In May and luly 1972, two large samples of speckled trout were collected for chlorinated hydrocarbon anal- ysis. The May collection of 58 females and 8 males averaged 38 cm in length and 500 g in weight. Testing showed that 29 percent of the females and 50 percent of the males in this collection contained DCPA in one or more tissues (Table 3). Overall, the highest levels were found in the liver tissues, although these were not always positive when other tissues were. The greatest amounts recorded for liver, ovaries, testes, and flesh were 196, 85, 200, and 26 ppb, respectively. The second sample of 9 females and 10 males averaged 38 cm in length and weighed 568 g. DCPA was not present in any of the tissues. No effort was made to deter- mine whether the two samples were from the same fish population. Two speckled trout (427 g and 257 g) col- lected during March 1973 also had DCPA residues. Several other species of fish (Table 1) were collected after the initial detection of DCPA in fish. Analysis of one mullet showed residues comparable to those found in menhaden of the same collection. Five samples of Rio Grande perch were collected at various locations along the Arroyo Colorado over a 4-month period. DCPA residues in the flesh varied from 0 to 217 ppb. One redfish had a trace of DCPA in the liver and testes. 56 Pesticides Monitoring Journai, TABLE 2. — DCPA residues in i 4 -in.-o.d. 3orosilicate glass packed with 10 percent OV-17 on 50/80-mesh Gas Chrom Q. The first two columns were Jsed for separating and analyzing ethion, azinphos- nethyl, and phosalone. The third column was used to leparate naled and dichlorvos; the fourth column was ised to separate ethion monooxon and ethion dioxon. Pemperatures of the injection part and detector for the Vlicro-Tek gas chromatograph were 240 C and 290-C, espectively. Injection and detection temperatures for he Varian Aerograph were 227^C and 230°C. re- spectively. Flow rates of the oxygen, air, hydrogen, and nitrogen carrier gases for the Micro-Tek were 20, 20, 200, and 90 ml/min, respectively. The Varian Aero- graph flow rates of the air, hydrogen, and nitrogen (carrier) gases were 170, 16.5, and 20 ml/min, respec- tively. Column temperatures for both in.struments for ethion, azinphosmethyl, and phosalone were held to 230°C; ethion monooxon and ethion dioxon samples were operated at 245'C. The naled and dichlorvos analyses were handled differently in that the column temperature was programmed with an initial tempera- ture of 130°C for 2 minutes following injection. Then it was programmed at a rate of 30°C/min until a final temperature of 170°C was reached; it was held at that temperature for 6 minutes before cooling to the initial temperature of 130°C. Results and Discussion Several extraction solvents including acetonitrile, ben- zene, chloroform, and 10 percent methanol in chloro- form were compared for the quantity of residue ex- tracted and the variability of the residue between individual samples. The quantity of the residue in the substrates was about the same regardless of the solvent selected; however, the 10 percent methanol in chloro- form gave results slightly more consistent than those produced by the other solvents. Extraction time of 30 minutes was also compared with another procedure (8) which included shaking the contents in 1:2 ratio of crop tissue to solvent for 2 minutes, letting it rest for 2 minutes, shaking for V2 minute, and resting V2 minute before transferring extract into a sample bottle. Again, residue levels were similar and well within the variations found between the replicates. However, there was a con- siderable degree of inconsistency between these sam- ples and those produced by the 30-minute extraction period. This study was concerned with maximum levels of resi- due that might be associated with surface exposure and includes residue levels that might be incorporated into the plant surface, as in the plant waxes or tissues just under the immediate surface. Therefore, residue levels reported here are probably higher than the actual surface residue to which a worker would be exposed in a field situation. Ascertaining how an individual is exposed to pesticides and which pesticides he is exposed to is very difficult. Much work needs to be done in this regard, but this was not the objective of the present study. Authors of this study had anticipated looking at some of the toxic degradation products which are more polar than their parent compounds. Therefore, it was necessary to extract with a mixture including a polar /OL. 8, No. 1, June 1974 61 solvent. Ten percent methanol in chloroform (2) was used throughout the study. Results from the fortified recovery studies ranged from 95 to 100 percent for all pesticides analyzed including the degradation products. The only two exceptions to this were ethion dioxon, which ranged from 80 to 95 percent recovery, and gutoxon, which could not be checked for a specific recovery because it was analyzed by thin-layer chromatography. Fortified extracts of gutoxon did look satisfactory even when fortified at 0.1 ppm. However, this was only a visual observation and quantitation was not attempted. Soil, leaves, cane, and bark were sampled periodically following each insecticide application. Harvested grapes were also sampled. Figures 2-4 show in graphic form the residue levels found on the various tissue samples, except for soil samples. Residue levels in the soil were relatively low, ranging from about 3.0 ppm on the first day of application to 1.0 ppm at harvest for azinphos- methyl and phosalone. Ethion had about the same initial residue but degraded rather rapidly, with a final residue at 42 days of 0.15 ppm. Naled had an initial level of about 0.5 ppm; the second sampling four days later showed less than 0.1 ppm except for one sample which had 0.2 ppm. Fruit samples of grapes were harvested and analyzed on two different dates, September 28 and October 8. Table 2 shows these residues and residues found on all tissues sampled at time of harvest. In all cases, residues found on the fruit were within the U.S. Government tolerance for each of the four pesticides surveyed. As mentioned earlier, insecticides were not applied to the five replicate plots (Rl-5) on the same day. The plot identified as Rl had its first application of ethion and azinphosmethyl on August 27, R2 and R3 on August 28, and R4 and R5 on August 31. The variability of residue between the three dates of application was less than the variability between samples with the same ap- plication on the same plot sampled the same day. Therefore, factors attributed to variation of application days were generally considered insignificant. The same was true of phosalone which was applied to Rl, R2, and R3 plots on September 11. R4 and R5 plots received their first application of phosalone on September 14. Naled was applied to all five plots on September 24. For each periodic sampling. Figures 2-5 show the high- est, lowest, and average residue for each pesticide per- FIGURE 2. — Residues of ethion. azinphosmeihyt. and phosalone extracted from grape leaves with an organic solvent wash from samples taken at time of application and through- out growing season. Each group of points represents range and mean of residues in samples after application 62 BARK AZINPHOSMETHYL* PHOSALONE ■ 1 20 30 DAYS 40 FIGURE 3. — Residues of ethion, azinphosmethyl, an phosalone extracted from grape bark with an organic solvei wash from samples taken at time of application and througl out growing season. Each group of points represents rang and mean of residues in samples after application Pesticides Monitoring Journa TABLE 2. — Insecticide residues found on grape tissues at time of harvest Insecticide Azinphosmethyl Ethion Phosalone Naled Tissue Analyzed Leaves Bark Cane Fruit Soil Leaves Bark Cane Fruil Soil Leaves Bark Cane Fruit Soil Leaves Bark Cane Fruit Soil Residues Found, ppm 7L3 28.2 6.60 2.561 LOO 5.80 8.20 0.40 0.70 I 0.15 98.8 47.0 6.80 L08 1 LOO <0.10 2.20 <0.10 <0.10 ' <0.10 ' Represents average of residues taken on two days of harvest- Sent 28 and Oct. F 80 60 ETHION AZINPHOSMETHYL PHOSALONE IGURE 4. — Residues of ethion, azinphosmethyl, and hosalone extracted from grape cane with an organic solvent ash from samples taken at time of application and through- ut growing season. Each group of points represents range and mean of residues in samples after application. 'OL. 8, No. 1, June 1974 TOTAL RESIDUE EXTRACTED FROM LEAVES, BARK. CANE 8 FRUIT 2 ND APPLICATION 3HD APPLICATION FIGURE 5. — Total insecticide residue extracted from leaves, bark, cane, and fruil of grape period from the initial time of the azinphosmethyl and ethion application. All reported naled residues include dichlorvos, even though the two insecticides were an- alyzed individually. Table 3 shows the average levels of residue found on the four separate substrates from initial day of application until harvest. Even though residue levels among samples varied considerably, levels in relation to time followed first-order kinetics, particularly in leaf tissues. Variability was much greater between cane and bark samples than between soil and leaf sam- ples. However, data did indicate that the rate of decline of each pesticide was considerably slower after the first few days, particularly for azinphosmethyl and phosalone. The irregularity between samples in the corky tissues can be explained by irregular absorption and adsorption and by exposure of the tissue to the spray. Irregularity is also explained by the variable exposure of the tissue to sunlight and other degradation processes. It is diffi- cult to get complete random sampling of old and new tissues from each plant because of the irregularities mentioned which may also explain the variability be- tween samples. In all cases, the stability of the insecticides in the bark and cane was very significant. Even 42 days after ap- plication, about half the original residue remained with the exceptions of naled and ethion. Naled was not detected in the cane; however, it was detected in the bark 14 days after application. Even though the residue level was relatively small, with about 15 percent of the initial deposit remaining at harvest time, the rate of decay was only 50 percent from day 4 to day 15. It appears that once the pesticide is in contact with the corklike tissue, it is absorbed readily and becomes quite stable over a long period of time. This may or may not be significant in relation to the exposure of field 63 TABLE -i.— Degradation of azinphosmelhyl, elhion, phosalone, and naled in grape plant tissues Average Residue on Leaves, ppm Average Residue Bark, ppm ON Average Residue on Cane, ppm Average Residue on Soil, ppm Days: INITIAL J, J z o u z z 2 o t X X z u z o J, J z z 2 < a APPLICATION i^5 o S a ss o tu zi in UJ z« < o s (DAYO) TO HARVEST < X o X < Z Is < H X 0. z < E < z a B z 0 322 225 48 13 10.4 7.5 2.8 1.4 3 283 156 38 22 10.6 4.0 1.6 1.4 4 271 139 38 22 8.1 4.5 3.7 2.4 7 251 84 34 19 6.9 3.1 3.1 1.5 14 167 49 310 41 17 106 7.5 1.4 13 1.5 0.56 2.9 17 150 37 310 47 15 92 6.8 1.6 12 2.3 0.44 3.5 21 132 28 252 30 10 65 6.9 1.3 13 1.1 0.20 2.5 24 92 16 182 37 7.6 55 4.8 0.85 10 1.3 0.35 1.7 27 98 12 210 36 35 8.9 80 14 6.6 0.90 11 I.l 1.6 0.28 2.1 0.27 28 69 10 144 <0.1 40 8.2 70 8.2 6.0 0.70 8.0 <0.10 1.7 0.40 1.8 <0.10 31 70 10 156 <0.1 26 8.2 62 5.6 5.7 0.60 10.5 <0.10 1.5 0.26 2.0 <0.10 42 71 5.8 99 <0.1 28 8.2 47 2.2 6.6 0.40 6.8 <0.10 1.0 0.15 1.0 <0.10 workers. Because high levels of residue are involved, however, the matter needs to be investigated. The two o.xygen analogs of ethion on grape leaves are shown in Table 4. All replicate samples analyzed for the metabolites were composited according to days from application. Residues of ethion monooxon and ethion dioxon were much lower than their parent compound the first few days following application. However, the relative amount of residues for the two products was much higher following the third day. The total amount of both ethion monooxon and ethion dioxon from the fourteenth day after application was approximately 50 percent of its parent compound and continued in this proportion throughout the growing season. Even though the percentage of these metabolites was quite high, the total amount of residue was much lower than other organophosphate pesticides studied. Gutoxon, the oxy- gen analog of azinphosmelhyl, was analyzed only the first 14 days following initial application due to the background on the thin-layer plates following the phosa- lone application. None of the samples analyzed showed any detectable residues of gutoxon or other cholinester- ase-inhibiting products in excess of 1 ppm for this period. TABLE 4. — Degradation of etiiion. etiuon monooxon. and ethion dioxon on grape leaves Days: Residue on leaves ppm application Ethion Monooxon (DAY 0) Ethion Ethion + TO harvest Ethion Monooxon Dioxon Ethion Dioxon 0 225 25 2.0 27 3 156 15 2.6 17.6 7 84 19 13 32 14 49 10 15 25 17 37 7.2 8.8 16 21 28 6.3 4.5 10.8 24 16 4.6 3.6 8.2 27 12 3.4 3.4 6.8 31 10 2.6 1.8 4.4 42 5.8 1.3 1.4 2.7 Table 5 and Figure 5 show the total residue less the i metabolites on the leaves, bark, cane, and fruit from i initial application until harvest. The day azinphosmelhyl i and ethion were applied, the average residue level was 626 ppm and after 14 days the sum of the residue in the plant dropped to about 300 ppm. On this day, phosalone was applied and the total residue rose to an. average level of 688 ppm. Just prior to the naled ap- plication 14 days later, the residue level should have- reached a level of about 350 ppm. according to projec- tion from the graph. The naled application did not have' a great effect on the total residue after the first day following application because of its rapid decay in most tissues. Perhaps the most significant fact displayed by this graph is the level of residue at or near time of harvest. This level approached nearly 300 ppm. about TABLE 5. Averages (ppm) of all four insecticides from initial application to harvest Days: initial application (day 0) to harvest Bark Cane Fruit Total Appli- cation FIRST APPLICATION 0 547 61 18 626 1 439 60 15 514 4 410 60 13 483 7 335 53 10 398 14 241 56 9.3 306 SECOND APPLICATION THIRD APPLICATION 27 328 144 22 494 28 223 126 15 12 376 31 236 102 17 14 369 42 175 85 14 4.4 278 64 Pesticides Monitoring Journ.a )ne half the initial and total deposit of azinphosmethyl ind ethion, and one third the highest residue level found )n any one sample at any given time. Even though this esidue level is high, it likely does not reflect the total neasure of hazard to which an agricultural worker is ■xposed because a considerable portion of these mate- ials were found in the bark and cane and, as mentioned ireviously, may not present an immediate hazard. This sped of pesticide residue in such tissues needs to be onsidered in a worker-safety study. Studies should also le conducted to ascertain whether the high residues ound in corklike tissue are, like leaf tissues, a potential lazard and, if so, in what manner and to what degree n individual would be exposed to them. LITERATURE CITED 0 Bailey, J. B.. D. Flalicrly. and D. Mengle. 1973. Pesticide residues on grape leaves evaluated for adverse effects on grape pickers. In manuscript. (2) Bowman, M. C, M. Bcrozci, and D. G. Leiick. 1968. Procedures for extracting residues of phosphorus insecti- cides and metabolites from field treated crops. J. Agr. Food Cheni. 16:796-802. (3) Oaks. D. M.. K. P. Dimick, and H. C. Hartmann. 1966. Aerograph Phosphorous Detector. Aerograph Research Notes, 1-13. (4) Hartmann, H. C. 1966. Phosphorus detector for pesticide analyses. Bull. Environ. Contam. Toxicol. 1:159-168. (5) Brady, S. S.. and J. E. Chancy. 1966. Flame photometric detector: application of a specific detector for phosphorus and for sulfur compounds sensitive to subnanogram quan- tities. J. Gas Chromatogr. 4(2):42-46. (6) Bowman. M. C. and M. Bcroza. 1968. Gas chromato- graphic detector for simultaneous sensing of phosphonis and sulfur containing compounds by flame photometry. Anal. Chem. 40:1448-1452. (7) Winlcrlin. W. L.. G. Walker, and H. Frank. 1968. Detec- tion of cholinesterase inhibiting pesticides following sep- aration on thin layer chromatograms. J. Agr. Food Chem. 16:808-812. 1 8) Magec, R. October 1970. Uniform method for determin- ing chemical degradation on tree foliage. Memorandum from Calif. Dept. of Agriculture. 3L. 8, No. 1, June 1974 65 APPENDIX Chemical Names of Compounds Discussed in This Issue ALDRIN AROCLOR 1260 AZINPHOSMETHYL (GuthionS) BHC (Benzene Hexachloride) CHLORDANE DCPA (Dacthal®) DDD DDE DDMU DDT DDVP DIBROM DICHLORVOS DIELDRIN ENDRIN ETHION GUTHION HEPTACHLOR HEPTACHLOR EPOXIDE LINDANE METHOXYCHLOR MIREX NALED NONACHLOR PHOSALONE POLYCHLORINATED BIPHENYLS (PCBs) RESMETHRIN TDE (DDD) TOXAPHENE ZOLONE Not less than 95% of 1,2,3,4, 10,10-hexachloro-l,4.4a,5,8,8a-hexahydro-I.4-fnrfo-pxo-5,8-dimethanonaphthalene PCB, approximately 60% chlorine 0.0-dimethyl 5[4-oxo-1.2,3-benzotria2in-3(4H ) ylmethyl] phosphorodithioate 1,2,3. 4.5 ,6-hexachlorocyLlohexane (mixture of isomers). Commercial product contains several isomers of which gamma is most active as an insecticide. l,2,3,5,6,7,8,8-octachloro-2,3,3a,4,7,7a-hexahydro-4,7-methanoindene. The technical product is a mixture of sev- eral compounds, including heptachlor, chlordene, and two isomeric forms of chlordane. Dimethyl-tetrachloroterephthalate See TDE. Dichlorodiphenyl dichloro ethylene. (Degradation product of DDT.) Main component: l,l-dichloro-2,2-bis(p-chlorophenyl) ethylene o.p'-DDE l.l-dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl) ethylene p,p'-DDE l.l-dichloro-2.2-bis(p-chlorophenyl) ethylene (l-chloro-2,2-bis|p-chlorophenyllethylene) a-bis(p-chlorophenyl )B.B, B-trichloroethane. Numerous isomers in addition to p.p'-DDT are possible, and some, are present in the commercial product. o,p'-DDT [l,l,l-trichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl) ethane] See dichlorvos. See naled. 2,2-dichlorovinyl dimethyl phosphate Not less than 85% of 1, 2.3,4, 10.10-hexachloro-6,7-epoxy-1.4,4a.5,6.7.8,8a-octahydro-l,4-fndo-f-TO-5.8-dimethano=i naphthalene 1,2,3,4, 10, 10-hexachloro-6.7-epoxy-l,4,4a,5,6,7,8,8a-octahydro-l,4-fni/o-fndo-5,8-dimethanonaphthalene 0,0,0',0'-tetraethyl S,S'-methylene bisphosphorodithioate See azinphosmethyl. l,4.5,6,7,8,8-heptachloro-3a,4,7.7a-tetrahydro-4,7-fndo-methanoindene 1,4,5,6,7,8,8-heptachloro 2,3-epoxy-3a,4,7,7a-tetrahydro-4,7-methanoindane Gamma isomer of benzene hexachloride ( 1,2,3.4. 5.6-hexachlorocyclohexane ) of 99 + % purity 1 .1 . l-Irichloro-2.2-bis( p-methoxyphenyl I ethane Dodecachlorooctahydro-l,3,4-metheno-lH-cyclobuta[cdlpentalene l,2-dibromo-2.2-dichloroethyl dimethyl phosphate Component of commercial quality chlordane 0,0-diethyl S-(6-chloro-2-oxo-benzoxazolin-3-yl) methyl phosphorodithioate Mixtures of chlorinated biphenyl compounds having various percentages of chlorine (5-benzyl-3-furyl methyl 2.2-dimeIhyl-3-( 2-methylpropenyl)cyclopropanecarboxylate 2,2-Bis (p-chlorophenyl)-l,l-dichloroethane (including isomers and dehydrochlorination products) Chlorinated camphene (67-69% chlorine); product is a mixture of polychlor bicyclic lerpenes with chlorinatO camphenes predominating See phosalone. 66 Pesticides Monitoring Journ; ERRATA PESTICIDES MONITORING JOURNAL, Volume 7, Number 1. In the paper "Pesticide Residues in Natural Fish Populations of the Smoky Hill River of Western iCansas— 1967-69," page 56, right column, third para- graph, second sentence, should read, "Dieldrin was detected in a small percentage (15%) of the samples; most of these had only trace «0.01 ppm) amounts." In Table 4 of the same paper, sample numbers and dieldrin residues for Station 5 were misquoted. Correct values for Station 5 appear below: TABLE 4.— Pesticide residues in fish and fish tissues from the Smoky Hill River. Kansas— 1967-69— Continued n = Program. It includes data on pesticide applications, soil residues, and crop residues col- lected from 1,506 cropland sites in 35 States. Pesticide ap- plication data are summarized hy all sites and by State. Soil residue data are itemized similarly, hut also incltide data by Topping region. Tables generally give the number of sites, number of times a pesticide was applied or detected, percent occurrence, arithmetic mean application rate or residue level, and range of residues detected. For .wme data, geometric means or 50 percentile levels, both with 95 percent confi- ience intervals, are presented. Pesticides applied most frequently were atrazine, 2,4-D. "aptan, and malathion. Farmers in cotton and corn cropping eginns applied the most pesticides: those in grass hay and mixed hay areas applied the least. Dieldrin, DDTR, aldrin, hlordane, and heptachlor epoxide were the chlorinated hy- Irocarbon residues found in soil most frequently. Highest •esidue levels were found in the corn and cotton cropping •■egions; lowest levels were in the hay, general farming, and •:nudl-grain soils. Introduction The National Soils Monitoring Program is an inte- ^al part of the National Pesticide Monitoring Program NPMP). The Program was initiated at the recommen- iation of the President's Science Advisory Committee -eport of 1964 entitled "Use of Pesticides." This com- nittee recommended that appropriate Federal agencies 'develop a continuing network to monitor residue levels n air, water, soil, man, wildlife, and fish" (/). The objective of NPMP is to determine levels and trends of pesticides and their degradation products in the various components of the environment (2). The initial goal, establishment of NPMP baseline or back- ground levels of pesticide residues, will provide a basis for comparison of subsequently identified pesticide residue levels in an environmental component and, ultimately, detection of significant trends. In determining levels and trends of pesticide residues, NPMP personnel have been guided by criteria of the Monitoring Panel of the Federal Working Group on Pest Management (2). The Panel has listed five areas of concern for individuals evaluating pesticide residue levels in environmental components. They are: any concentration of a pesticide known to be potentially harmful; increasing trends; levels exceeding standards; recognition of adverse effects on humans; and erratic variability, a statistically oriented observation that is potentially common to each stratum sampled. This report is a summary of the pesticide application data and soil and crop residue data collected in Fiscal Year 1970 (FY-70) from 1.506 sites located in 35 States. Data were not collected from certain agricultural States because of budgetary limitations. States omitted were those abounding in wheat and other grains, which require fewer pesticides than do many nongrain crops. The FY-69 (i) and FY-70 findings have achieved the initial NPMP goal of establishing baseline data. Authors will identify significant trends as more data arc accumu- lated. Pollutant Pattiways Branch. Monilorint^ .Systems. Research and Devel- opment Laboratory, National Environmental Research Center, U.S. Environmenlal Protection Agency. Las Vesas. Nev. Ecological Monitoring Branch. Technical Services Division, Office of Pesticides Programs. U.S. Environmental Protection Agency. Missis- sipri Tesi Facility. Miss. Plant Protection and Quarantine Programs. Animal and Plant Health Inspection Service. U.S. Department of Agriciilliire. H\altsvdle. Md. Ecological ^fonitoring Branch. Technical Services Division. Office of Pesticides Programs. U.S. Environmental Pnileclion Agency. Wash- ington. DC. Samplirig Sampling techniques involved in this study were gen- erally the same as those employed by Wiersma, Sand, and Cox in a 1971 study {4). In FY-70, cropland sites in 35 States were sampled (Fig. 1). This included all but the following States: Alaska, Arizona, Colorado, Hawaii, Idaho, Kansas, Montana, Nevada, New Me.x- /OL. 8. No. 2, September 1974 69 KEY JSyM States sampled NOTE: in the F Y-70 study, smaller Eastern States were combined to obtain a more reasonable sample. Such State groups represent unmarked areas on the maps. They are: -Mid-Atlantic: Delaware, Maryland, and New Jersey -New England: Connecticut, Maine. Massachusetts, New Hampshire, Rhode Island, and Vermont —Virginia and West Virginia FIGURE 1. Stales sampled for pesticide residues, FY -70 ico, North Dakota, Oregon, Texas, Utah, Washington, and Wyoming. Although an attempt was made to collect soil and crop samples just prior to harvest, this was not always possible. Crop samples were not collected if the crop was not mature and/ or ready for harvest. A nalytical Procedures SOIL To analyze soil for pesticide residues, a 300-g sub- sample was taken from a thoroughly mixed field sam- ple. The subsample was dampened with water and ex- tracted with 600 ml of 3:1 hexane:isopropanol solvent by concentric rotation for 4 hours. The isopropanol was removed by three distilled water washes and the hexane extract was dried through anhydrous sodium sulfate. The sample extract was then stored at low temperature for subsequent gas-liquid chromatographic analysis. CROPS To analyze crop samples containing less than 2 per- cent fat (e.g., alfalfa, bur clover, corn stalks, cotton stalks, green bolls, and miscellaneous hay), technicians blended 100-g samples of the crop with 25 ml of dis- tilled water for 3 minutes in SOO ml of acetonitrile. Half the sample extract, containing 50 grams of tl original sample, was decanted into a 500-ml Erlenmey flask. After concentration under a three-ball Snyd column to approximately 10 ml, 100 ml of hexane w added, and the hexane-acetonitrile azeotrope was aga concentrated to 10 ml. This process was carried o three times to remove essentially all the acetonitril The hexane extract was dried through anhydrous s dium sulfate, the volume was adjusted to 100 ml, ai the extract was stored at low temperature until rea( for partitioning. To analyze crop samples containing more than percent fat (e.g.. corn kernels, cottonseeds, and so beans), researchers prewashed a 100-g sample with li ml of isopropanol and then with 100 ml of hexar Both prewashes were discarded. The sample was drie dry-blended, added to 100 ml isopropanol, and blend again. After the addition of 300 ml of hexane, the is propanol was removed by two washes with aqueo NaCl solution and one wash with distilled water. T water-alcohol layers were discarded; the hexane lay was concentrated, adjusted to 100 ml, and held at k temperature for partitioning. After extraction, all crop samples were partition with hexane-acetonitrile as follows: a 50-ml portion 70 Pesticides Monitoring Journ he hexane sample extract was shaken with 100 ml of cetonitrile in a 500 ml separatory funnel. The bottom cetonitrile layer was set aside. Nanograde acetonitrile 100 ml) was added to the hexane extract and the sep- ration step described above was repeated two more imes; then the hexane was discarded and the three cetonitrile layers were combined. The 300-ml acetoni- rile extract, which contained essentially all the pesti- ides in the original hexane extract, was backwashed /ith 25 ml of acetonitrile-saturated hexane and the lexane layer was discarded. The acetonitrile sample xtract was concentrated to approximately 10 ml under . three-ball Snyder column, and 100 ml of hexane was dded. Addition of hexane and concentration to approx- mately 10 ml was performed three times, after which he sample was essentially in hexane. Remaining hexane xtract was diluted to appropriate volume and held at aw temperature for subsequent florisil column cleanup *nd fractionation. rAS-LIQUID CHROMATOGRAPHY Analyses were performed on gas chromatographs quipped with tritium foil electron affinity detectors for Tganochlorine comp>ounds and thermionic or flame hotometric detectors for organophosphorous com- lounds. A multiple-column system employing polar and lonpolar columns was utilized to identify and confirm iiesticides. Instrument parameters were as follow: Columns: Glass, 6 mm o.d. by 4 mm i.d., 183 cm long, packed with one of the fol- lowing: 9 percent QF-1 on 100/120 mesh Gas-Chrom Q; 3 percent DC- 200 on 100/120 mesh Gas-Chrom Q; or 1.5 percent OV-17/I.95 per- cent QF-1 on 100/120 mesh Sepul- copwrt. Carrier Gases: 5 percent methane-argon at a flow rate of 80 ml/min; prepurified nitro- gen at a flow rate of 80 ml/min. Temperatures: Detector 200°C Injection port 250°C Column QF-1 166°C Column DC-200 170-175''C Mixed column I85-190°C ■ensitivity or minimum detection levels for organo- hlorine compounds ranged from 0.002 to 0.03 ppm xcept for combinations of polychlorinated biphenyls PCB's), chlordane, toxaphene, and other chemicals «hich had minimum detectable levels of 0.05 to 0.1 pm. Minimum detectable levels for organophosphorous ompounds were approximately 0.01 to 0.03 ppm. /hen necessary, residues were confirmed by thin-layer hromatography or p-values. RECOVERY STUDIES For organochlorine pesticides, average recovery rate in soil was 90 to 110 percent. Recovery values for stalks and hay ranged from 80 to 95 percent, with an average of 89 percent; corresponding values for grains were 90 to 100 percent, with a 95 percent average. For organophosphate pesticides, average recovery values were 67.1 percent for soybeans, 86 percent for sorghum grain, and 60 percent for corn stalks. Residue levels in both crops and soils are expressed on a dry-weight basis and are corrected for percent recovery. Results Tables presented in this report are divided into three groups: pesticides applied to cropland, chlorinated hy- drocarbon pesticide residues in cropland soil, and chlor- inated hydrocarbon and organophosphate pesticide res- idues in crops. Pesticide application data are further subdivided by all sites and by State. Residues in soil are summarized by all sites and by States and cropping regions. Most tables list the number of samples, the number of times a pesticide was applied or detected, percent occurrence of that particular pesticide, arithmetic mean, and range. Readers should exercise caution when inter- preting the arithmetic mean because data are not nor- mally distributed. The arithmetic mean tends to be con- siderably higher than the corresponding median; but because of the skewed distribution of these data, the mean may not be a good indication of central tendency. Hence geometric means and 50 percent estimates have been presented. PESTICIDES APPLIED TO CROPLAND When soil samples were collected from cropland, field personnel attempted to contact farmers to determine what pesticides had been applied to the sites during the year of sampling. These data were not always available: only 1,346 use records were collected from 1,506 sites. Tables summarizing application data show the number of sites using a pesticide, percent of sites using a par- ticular pesticide, average rate of application for sites using the pesticide, and average amount of pesticide applied to all sites. The latter figure was determined by dividing the total amount of active ingredients applied to all sites by the total number of sites surveyed. All rate data are expressed in pounds per acre (lb/ a). To convert to kilograms per hectares (kg/ ha), multiply by 1.1208. ALL SITES Table 1 is a summary of the 1,346 sites surveyed from 35 States. The most common pesticides were atra- zine, 2,4-D, captan, and malathion, which were used 'oL. 8, No. 2, September 1974 71 on 11, 9, 8, and 6 percent of the sites, respectively. Aldrin and DDT were used on 4 percent of the sites. BY STATE Table 2 is a Because some few samples, representative tic: Delaware, land: Connect shire, Rhode West Virginia. breakdown of the all-site data by State. of the smaller Eastern States had very several were combined to obtain more data. State groups used were Mid-Atlan- Maryland, and New Jersey; New Eng- icut, Maine, Massachusetts, New Hamp- Island, and Vermont; and Virginia and Considering the number of States sampled, it is not feasible to discuss pesticide applications of each State. It is, however, worthwhile to mention application fre- quencies of a few States. Because Mississippi farmers produce large quantities of cotton, a crop which demands heavy pesticide treat- ment, they apply a large number of pesticides to much of their cropland. Growers used the following pesticides most frequently on the specified percentage of sites: methyl parathion, 48 percent; trifluralin, 42 percent; DDT, 32 percent; Cotoran (fluometuron), 26 percent; toxaphene, 23 percent; Bidrin (dicrotophos). 19 percent; MSMA, 19 percent; folex, 16 percent; and diuron, 13 percent. In Illinois, a major corn-growing State, pesti- cides applied most frequently and the percentage of sites treated were: captan, 45 percent; malathion, 45 percent; Ramrod (propachlor), 24 percent; aldrin, 18 percent; atrazine, 16 percent; and heptachlor, 12 per- cent. In contrast to these States were Oklahoma, Min- nesota, Kentucky, and the New England group, where no pesticide was applied to more than 8 percent of the sampling sites. Herbicide 2,4-D was applied most frequently in South Dakota; of the sites sampled, 25 percent were treated. Atrazine was applied most frequently in Wis- consin and North Carolina: 25 and 26 percent of the sites, respectively, were treated. Captan was applied most frequently in the Mid-Atlantic States; 47 percent of the sites were treated. BY CROP Table 3 presents pesticide application data for sev- eral crops. The few pesticides applied to alfalfa and bur clover were not widely used. Of the many different pesticides applied to field corn, atrazine, captan, 2,4-D, and malathion were employed most frequently. Farmers used 45 different pesticides on cotton fields; of these compounds, 14 were applied to more than 10 percent of the sites. In contrast, field workers applied only two pesticides to both grass hay and mixed hay fields. Al- though 27 pesticides were applied to soybeans, only amiben and trifluralin were applied to more than 5 per- cent of the sites sampled. CHLORINATED HYDROCARBON PESTICIDE RESIDUES IN CROPLAND SOIL All 1,506 soil samples from the 35 States were ex amined for chlorinated hydrocarbon residues. A fev samples were analyzed for additional pesticides, bu these data were insufficient for consideration. In thi future, soil residue analyses will be expanded to includi more compounds. Samples were not analyzed for PCB's because Uttlt' was known about them at the time of analysis. A reviev of the gas chromatograph traces revealed that PCB res idues were seldom present and, when they were, th( residues existed only in very small quantities. PCB peak; were recognized and not confused with DDT. ALL SITES Table 4 summarizes chlorinated hydrocarbon soil res) idue data for all States, including the number of time a pesticide was detected, percent occurrence, arithmeti< mean, geometric mean with 95 percent confidence inter val, and range of detected residue values. The geometrii mean was used because it gives a better indication o the central tendency of the data. It was calculated usinj a 0.01 addition to eliminate the problem of zeros and was later subtracted from the calculated mean. The most widely distributed chlorinated hydrocarbon was dieldrin: 31 percent of the soil samples had detect able residues. It was followed by DDTR (DDE-f-TDE) aldrin, chlordane, and heptachlor epoxide, which wen detected at 23, 13, 11, and 10 percent of the sites respectively. DDTR existed in the highest concentration, with ai arithmetic mean of 0.30 ppm and geometric mean o 0.0116 ppm. Although the arithmetic mean for dieldrij was much lower than that for DDTR, the geometric means are indistinguishable at the 95 percent confldeno level. The very high residue value of 113 ppm as show] in the range of DDTR can have a major effect on ai arithmetic mean. The effect of extreme values upoi the geometric mean is considerably less. BY STATE Chlorinated hydrocarbon residues in soils of specifi States (Table 5) are presented in the same manner a the all-sites data except for the geometric means. Comparisons of the percent occurrence of aldrir dieldrin, heptachlor epoxide, DDTR, and chlordane ar presented in Figures 2 through 6. The key for eacl figure is based on the arithmetic average percent occur rence (x) of the given pesticide for all sites. The fou categories used are: greater than 2x, greater than x bu less than 2x, greater than V2X but less than x", and les than Vix". Iowa and Illinois showed the highest percent occui rence of aldrin, dieldrin, and heptachlor epoxide (Fif 72 Pesticides Monitoring Journa; X « 13.48%. (Average % occurrence of aldrin for all sites.) 2x ^R^ / / \ t— t*M < 1 ^"i ■" ^ ^'^x^ ;. ^^M rm ^J V----- 3V-^^S^5 V Y 1 1 '°^* Kill. ^ ^ ^ S^/-^'-^^ ■""•^ ^ r^r' t; T "^> r* ■3 IL^ 'i.i^ ' "' V J' "■ k_ 1 ^a«>k '^ y KEY ^^>^, , . , .—A U— *s^ IS>— .^ X = 30.88%. (Average % occurrence of dieldrin for all sites.) "^V V \ x<^<2x \ ^ '^ i/?x TABLE 2. Pesticides applied to cropland by State, FY-70 Pesticide No. Sites Using Pesticide Percent Sites Using Pesticide Arith. Mean Appl. Rate (LB/A)1 Alabama, 22 Sitbs Arkansas, 47 Sites Arith. Mean (LB/A)1 Azinphosmethyl 4.55 4.00 0.1818 Azodrin 4,55 12.00 0.5455 Copper sulphate 4.55 8.00 0.3636 Cotoran 9.09 1.50 0.1364 DDT 27.27 11.08 3.0227 OEF 4.55 1.50 0.0682 DSMA 4.55 2.00 0.0909 Endothall 4.55 0.90 0.0409 Endrin 4.55 10.50 0.4773 ^Jilralin 9.09 1.00 0.0909 Parathion. ethyl n.64 16.06 2.1900 Parathion, methyl 31.82 6.54 2.0823 5trobane 4.55 3.00 0.1364 Sulfur 4.55 72.00 3.2727 foxaphene 13.64 48.31 6.5873 frilluralin 13.64 2.03 0.2773 \ldrin 1 2.13 0.01 0.0002 \miben 2 4.26 1.00 0,0428 Azodrin 1 2.13 3.75 0,0798 Taptan 2 4.26 0.03 0,0015 Tarbaryl 3 6.38 1.70 0,1085 Ceresan M 2 4.26 0.05 0,0023 >resan red 2 4.26 0.10 0,0043 rhloroneb 3 6.38 0.01 0,0009 Totoran 2 4.26 0.77 0,0330 Dalapon 2.13 3.00 0,0638 !,4-DB 4.26 0.50 0,0213 DDT 4.26 6.25 0,2660 DEF 2.13 0.75 0,0160 Dinitrobutylphenol 2.13 10.00 0.2128 DinitTOcyclohexylphenoI 2.13 0.50 0.0106 DSMA 2.13 1.00 0.0213 5PN 2.13 0.50 0.0106 Ethylene dibromide 2.13 0.01 0.0002 -olex 4.26 1.50 0.0638 _asso 2.13 0.50 0.0106 -inuron 2.13 1.50 0.0319 vlethylmercury dicyandiamide 6.38 0.01 0.0006 vlelhyl trilhion 2.13 5.50 0.1170 vlonuron 2.13 0.25 0.0053 vISMA 6.38 3.33 0.2128 "Jitralin 4.26 0.44 0.0187 •4orea 2.13 1.00 0.0213 ^araquat 2.13 0.25 0.0053 'arathion, methyl 10 21.28 4.00 0.8511 ^horate 2.13 0.50 0.0106 ^rometryne 2.13 0.75 0.1600 'ropanil 2.13 2.50 0.0532 foxaphene 2 4.26 1.75 0.0745 Frifluralin 9 19.15 0.68 0.1311 California, 43 Sites Azinphosmethyl 3 6.98 0.92 0.0640 \zodrin 2 4.65 4.38 0.2035 Jidrin 2.33 0.75 0,0174 Bordeaux mixtures 2.33 1.00 0,0233 :arbaryl 2.33 1.25 0,0291 rarbophenothion 2.33 0.50 0,0116 ;hlorobenzilate 2.33 1.00 0,0233 :,4-D 2.33 0.88 0,0205 DAC 2.33 4.00 0,0930 JDT 6.98 3.00 0,2093 JEF 2.33 3.00 0,0698 Diazinon 2.33 2.00 0,0465 >icofol 4.65 0.88 0,0407 Jimethoate 2.33 0.33 0,0077 Moxathion 2.33 0.75 0,0174 )isulfoton 2.33 1.00 0.0233 DSMA 2.33 2.00 0.0465 indosulfan (I) 4.65 0.88 0.0407 :ndrin 2.33 0.75 0.0174 :thion 4.65 1.50 0.0698 enthion 2.33 0.09 0.0021 'lalathion 4.65 2.02 0.0942 -Icthyl demeton 2.33 0.25 0.0058 'araquat 4.65 0.18 0.0086 'arathion, ethyl 16.28 0.99 0.1616 'arathion, methyl 2.33 0.10 0.0023 imazine 2 4.65 3.30 0.1535 odium chlorate 3 6.98 8.17 0.5698 Continued next page) /OL. 8, No. 2, Septembi =R 1974 79 TABLE 2 (cont'd). Pesticides applied to cropland by State, FY -70 Pesticide No. Sites Using Pesticide Percent Sites Using Pesticide Arith. Mean Appl. Rate (LB/A)1 Awth. Mean (LB/A)1 Sulfur Tetradifon Toxaphene Trifluralin 5 1 3 1 11.63 2.33 6.98 2.33 41.71 0.75 6.00 2.00 4.8502 0.0174 0.4186 0.0465 Florida, 18 Sites Aldrin 5.56 4.44 0.2467 Atrazine 5.56 0.50 0.0278 Carbaryl 5.56 5.00 0.2778 Carbophenothlon 11.11 1.13 0.1250 Chlorobenzilate '11.11 0.92 0.1022 Copper oxide 11.11 4.65 0.5167 2,4-D 5.56 2.00 O.Ull Dalapon 5.56 2.00 0.1111 Dicofol 5.56 2.00 0.1111 Dimethoate 5.56 6.10 0.3389 Ethion 27.78 2.30 0.6389 Lead arsenate 5.56 6.40 0.3556 Parathion, ethyl 11.11 3.55 0.3944 Sulfur 22.22 60.65 13.4778 2,4,5-T 5.56 2.00 0.1111 Toxaphene 5.56 4.00 0.2222 Georgia, 30 Sites Atrazine 2 6.67 2.70 0.1800 Azodrln 1 3.33 0.60 0.0200 Benefin 2 6.67 1.31 0.0873 Bidrin 1 3.33 0.30 0.0100 Calcium arsenate 1 3.33 9.80 0.3267 Captan 8 26.67 0.05 0.0147 Carbaryl 5 16.67 4.45 0.7417 Ceresan red 4 13.33 0.01 0.0013 Copper sulfate 8 26.67 62.73 16.7283 DDT 7 23.33 3.96 0.9233 Dinitrobutylphenol 2 6.67 1.50 0.1000 Disulfoton 1 3.33 0.80 0.0267 Diihane M-45 1 3.33 1.50 0.0500 DSMA 1 3.33 0.50 0.0167 Endrin 1 3.33 0.33 0.0110 Falone 1 3.33 2.00 0.0667 Linuron 1 3.33 1.00 0.0333 Malathion 4 13.33 0.01 0.0013 Methoxychlor 4 13.33 0.03 0.0040 Methyl trithion 1 3.33 10.00 0.3333 Mirex 1 3.33 0.01 0.0003 Parathion, methyl 2 6.67 1.02 0.0683 Simazine 1 3.33 2.50 0.0833 Sulfur 3 10.00 20.00 2.0000 Toxaphene 7 23.33 4.63 1.0800 Trifluralin 1 3.33 1.50 0.0500 Vernolate 5 16.67 2.20 0.3667 Illinois, 51 Sites 2 Aldrin 9 17.65 0.92 0.1627 Amiben 5 9.80 2.60 0.2549 Atrazine 8 15.69 1.94 0.3049 Bux-ten (bux) 4 7.84 2.97 0.2333 Captan 23 45.10 0.03 0.0122 Carbaryl 1 1.96 2.00 0.0392 Ceresan red 2 3.92 0.01 0.0004 2.4-D 3 5.88 0.31 0.0182 DAC 1 1.96 1.50 0.0294 Dalapon 1 1.96 1.00 0.0196 Diazinon 2 3.92 0.50 0.019.S Dicamba 1 1.96 0.02 0.0004 Furadan 1 1.96 1.00 0.0196 Heptachlor 6 11.76 0.84 0.0984 Lasso 1 1.96 1.40 0.0275 Linuron 1 1.96 1.00 0.0196 Malathion 23 45.10 0.01 0.0045 Methoxychlor 5 9.80 0.01 0.0010 Phorate 1 1.96 0.05 0.0010 Ramrod 12 23.53 1.23 0.2904 Silvex 1 1.96 0.50 0.0098 Vernolate 1 1.96 1.30 0.0255 Indiana, 77 Sites Aldrin 7 9.09 0.82 0.0742 Amiben 6 7.79 0.97 0.0756 Atrazine 13 16.88 1.37 0.2318 Captan 11 14.29 0.01 0.0016 Ceresan red 1 1.30 0.02 0.0003 2,4-D 9 11.69 0.66 0.0769 (Conlinued next page) 80 Pesticides } vIONITORING JOURNA TABLE 2 (cont'd). Pesticides applied to cropland by State, FY-70 Pesticide Dinitrobutylphenol Heptachlor Lasso Linuron Malathion Methoxychlor Methylmercury dicyandiamide MSMA NPA Ramrod Trifluralin Amiben Atrazine Chloroxuron 2,4-D Dalapon No. Sites Using Pesticide Percent Sites Using Pesticide 1.30 2.60 6.49 2.60 7.79 2.60 1.30 1.30 1.30 5.19 1.30 Kentucky, 31 Sites 6.45 6.45 3.23 6.45 3.23 Michigan, 34 Sites Arith. Mean Appl. Rate (LB/A)1 2.00 0.76 1.40 1.05 0.01 0.01 0.01 4.00 4.00 2.06 1.00 2.00 1.75 2.00 0.88 1.00 Arith. Mean (lb/a)' 0.0260 0.0199 0.0909 0.0273 0.0008 0.0003 0.000 1 0.0519 0.0519 0.1071 0.0130 Iowa, 152 Sites Aldrin 15 9.87 0.83 0.0820 Amiben 14 9.21 0.84 0.0776 Atrazine 24 15.79 1.50 0.2367 Bux-ten (bux) 8 5.26 0.99 0.0520 Captan 2 1.32 0.01 0.0001 Carbaryl 1 0.66 1 60 0 0105 Chevron RE-5353 1 0.66 0.70 0 0046 CIPC 1 0.66 0.66 0.0043 2,4-D 17 11.18 0.60 0.0669 2,4-DB 2 1.32 0.88 0.0115 Diazinon 3 1.97 0.50 0.0099 Fensulfothion 2 1.32 1.38 0.0181 Heptachlor 5 3.29 0.61 0.0199 Lasso 3 1.97 0.91 0.0180 Lindane 1 0.66 0.02 0.0001 Linuron 1 0.66 0.60 0.0039 1 0.66 1.00 0 0066 Parathion. ethyl 1 0.66 0.02 0.0001 Phorate 7 4.61 0.85 0.0391 Ramrod 11 7.24 1.37 0.0991 Sutan 2 1.32 4.50 0.0592 Trifluralin 3 1.97 0.40 0.0079 0.1290 0.1129 0.0645 0.0565 0.0323 Louisiana, 25 Sites Aldrin 3 12.00 0.10 0.0124 Azinphosmethyl 3 12.00 1.18 0.1420 Buturon 2 8.00 0.01 0.0008 Captan 1 4.00 0.02 0.0008 Ceresan L 2 8.00 0.01 0.0008 Cotoran 1 4.00 1.00 0.0400 2,4-D 4 16.00 0.88 0.1400 Dalapon 3 12.00 3.67 0.4400 2,4-DB 1 400 0.75 0.0300 DDT 2 8.00 2.50 0.2000 DEF 1 4.00 1.12 0.0448 Diuron 1 4.00 0.70 0.0280 DSMA 1 4.00 6.00 0.2400 Norea 1 4.00 1.00 0.0400 NPA 1 4.00 2.00 0.0800 Parathion, methyl 6 24.00 3.54 0.8500 Propanil 2 8.00 3.00 0.2400 Silvex 2 8.00 0.75 0.0600 TCA 2 8.00 5.25 0.4200 TDE 1 4.00 6.00 0.2400 Terbacil 2 8.00 0.39 0.0316 Toxaphene 3 12.00 6.33 0.7600 Trifluralin 4 16.00 0.81 0.1300 Atrazine 4 11.76 3.38 0.3971 Azinphosmethyl 3 8.82 3.75 0.3309 Captan 2 5.88 13.00 0.7647 Carbaryl 2 5.88 15.00 0.8824 CDEA 1 2.94 2.00 0.0588 Chlordane 1 2.94 1.00 0.0294 2,4-D 3 8.82 1.17 0.1029 Dodine 2 5.88 2.22 0.1309 EPTC 2 5.88 3.50 0.2059 Ethion 1 2.94 2.50 0.0735 Ferbam 1 2.94 4.60 0.1353 Malathion 1 2.94 3.75 0.1103 Simazine 1 2.94 2.00 0.0588 Sulfur 2 5.88 97.50 5.7353 'Continued next page) Vol. 8, No. 2, September 1974 81 TABLE 2 (cont'd). Pesticides applied to cropland by State, FY-70 Pesticide No. Sites Using Pesticide Percent Sites Using Pesticide Arith. Mean Appl. Rate (LB/A)1 Arith Mean (LB/A)1 Mid-Atlantic States.' 15 Sites Aldrin 1 6.67 1.20 0.0800 Atrazine 2 13.33 1.28 0.1707 Azinphosmethyl 6.67 0.05 0.0033 Captan 46.67 0.02 0.0107 2,4-D 20.00 0.77 0.1533 EPTC 6.67 100.00 6.6667 Heptachlor 6.67 0.04 0.0027 Lasso 6.67 0.20 0.0133 Linuron 6.67 0.15 0.0100 Malathion 26.67 0.01 0.0027 Maneb 6.67 2.00 0.1333 Melhoxychlor 13.33 0.01 0.0013 Methyl demeton 6.67 2.00 0.1333 Phorate 6.67 20.00 1.3333 Simazine 6.67 2.20 0.1467 Minnesota, 119 Sites Amiben 4 3.36 2.00 0.0672 Atrazine 6 5.04 1.90 0.0958 Bux-ten (bux) 1 0.84 4.00 0.0336 Captan 2 1.68 0.01 0.0002 Carbaryl 2 1.68 1.45 0.0244 2,4-D 10 8.40 0.74 0.0626 Dalapon 1 0.84 1.00 0.0084 Isopestox 1 0.84 0.01 0.0001 Lasso 1 0.84 1.00 0.0084 Linuron 1 0.84 2.50 0.0210 Malatliion 1 0.84 0.01 0.0001 MCPA 7 5.88 0.86 0.0504 Piiorate 2 1.68 10.50 0.1765 Ramrod 7 5.88 1.94 0.1143 Silvex 1 0.84 0.40 0.0034 Mississippi, 31 Sites Azinpliosmethyl I 3.23 3.00 0.0968 Bidrin 6 19.35 0.23 0.0452 Captan 1 3.23 0.01 0.0003 Carbaryl 2 6.43 4.50 0.2903 Ceresan red 3 9.68 0.01 0.0010 Chloroneb 2 6.45 0.01 0.0006 Cotoran 8 25.81 0.38 0.0977 DDT 10 32.26 6.22 2.0081 DEF 2 6.45 1.13 0.0726 Dinitrobutylphenol 3.23 0.52 0.0168 Dinitrocresol 6.45 0.10 0.0068 Disulfoton 9.68 0.02 0.0016 Diuron 12.90 0.13 0.0171 DSMA 3.23 0.62 0.0200 Endrin 3.23 1.65 0.0532 EPN 3.23 1.70 0.0548 Folex 16.13 1.22 0.1968 Lasso 3.23 0.35 0.0113 Linuron 6.45 0.79 0.0510 Monuron 3.23 0.38 0.0123 MSMA 19.35 0.71 0.1384 Nitralin 6.45 0.88 0.0565 NPA 3.23 0.26 0.0084 Paraquat 9.68 0.16 0.0152 Parathion, methyl 15 48.39 4.82 2.3342 PCNB 3.23 0.01 0.0003 Sodium chlorate 3.23 3.00 0.0968 Toxaphene 22.58 11.86 2.6774 Trifluralin 13 41.94 0.98 0.4113 Missouri, 81 Sites Aldrin Amiben Atrazine Carbaryl 2,4-D Diazinon Diuron Heptachlor Linuron Ramrod Sutan Trifluralin 10 5 17 1 9 1 3 1 2 3 1 5 12.35 6.17 20.99 1.23 11.11 1.23 3.70 1.23 2.47 3.70 1.23 6.17 1.00 1.35 1.83 2.00 0.58 0.60 0.83 1.00 0.50 1.52 4.00 0.85 0.1235 0.0833 0.3840 0.0247 0.0648 0.0074 0.0309 0.0123 0.0123 0.0562 0.0494 0.0525 Nebraska, 106 Sites Aldrin Amiben Atrazine 1 2 16 0.94 1.89 15.09 0.50 0.63 1.28 0.0047 0.0118 0.1937 (Continued next page) 82 Pesticides Monitoring Journal TABLE 2 (cont'd). Pesticides applied to cropland by Slate, FY-70 No. Percent AwTH. Mean Awth. Sites Usino Sites Using Appl. Rate Mean Pesticide Pesticide Pesticide (LB/A)> (LB/A)1 Bux-ten (bux) 2 1.89 1.22 0.0231 Caplan 18 16.98 0.01 0.0017 Carbaryl 1 0.94 1.60 0.0151 Ceresan L 1 0.94 0.10 0.0009 2.4D 9 8.49 0.60 0.0509 Dalapon 1 0.94 2.50 0.0236 Demeton 1 0.94 0.25 0.0024 Diazinon 1 0.94 0.22 0.0021 Dicldrin 1 0.94 0.01 0.0001 Disulfoton 3 2.83 1.14 0.0324 Endrin 2 1.89 0.22 0.0042 Fensulfolhion 2 1.89 1.03 0.0195 Heptachlor 2 1.89 0.01 0.0002 Malalhton 18 16.98 0.01 0.0017 Melhoxychlor 2 1.89 0.01 0.0002 Melhylmercury dicyandiamide 1 0.94 0.01 0.0001 Phorate 6 5.66 0.95 0.0540 Ramrod 4 3.77 1.09 0.0410 Thiram 3 2.83 0.01 0.0003 New England,' 18 Sites No pesticides applied New York, 35 Sites Atrazine 14.29 2.00 0.2857 Azinphosmethyl 2.86 37.50 1.0714 Caplan 14.29 30.01 4.2869 Carbaryl 8.57 8.33 0.7143 2.4-D 11.43 0.63 0.0726 Demeton 2.86 1.50 0.0429 Dichlorprop 2.86 0.50 0.0143 Dicofol 2.86 4.04 0.1154 Dieldrin 2.86 0.02 0.0006 Dodine 2.86 11.25 0.3214 EPTC 2.86 3.00 0.0857 Malalhion 11.43 0.01 0.0011 Melhoxychlor 8.57 0.01 0.0009 Melhylmercury dicyandiamide 5.71 <0.01 0.0003 Nitrale 20.00 51.71 10.3429 Sutan 2.86 3.75 0.1071 Thiram 2.86 0.10 0.0029 North Carolina, 31 Sftes Amiben 1 3.23 2.00 0.0645 Atrazine 8 25.81 2.20 0.5677 Azodrin 1 3.23 0.60 0.0194 Caplan 1 3.23 0.01 0.0003 Carbaryl 3 9.68 1.32 0.1274 Ceresan red 2 6.45 0.10 0.0065 Copper oxide 1 3.23 1.80 0.0581 Copper-8-quinolinolate 1 3.23 0.01 0.0003 2,4-D 3 9.68 0.83 0.0806 DDT 4 12.90 1.60 0.2065 Diazinon 2 6.45 3.58 0.2310 Dichloropropane 1 3.23 26.00 0.8387 Dichloropropene 2 6.45 64.21 4.1426 Dinitrobulylphenol 1 3.23 1.50 0.0484 Diphenamid 2 6.45 3.00 0.1935 Endosulfan (I) 1 3.23 0.08 0.0026 Lindane 1 3.23 0.01 0.0003 Malathion 1 3.23 2.50 0.0806 Maleic hydrazide 3 9.68 1.80 0.1742 MSMA 2 6.45 2.38 0.1532 Parathion, ethyl 2 6.45 0.60 0.0387 Sulfur 1 3.23 33.75 1.0887 TDE 4 12.90 1.33 0.1710 Thiram 1 3.23 0.01 0.0003 Toxaphene 1 3.23 2.00 0.0645 Ohio, 69 Sites (Continued next page) Vol. 8, No. 2, September 1974 Aldrin 6 8.70 1.67 0.1449 Amiben 8 11.59 1.55 0.1801 Atrazine 12 17.39 1.70 0.2955 Captan 1 1.45 0.01 0.0001 CDAA 1 1.45 0.60 0.0087 2,4-D 9 13.04 0.71 0.0920 Dicamba 1 1.45 0.06 0.0009 Dieldrin 1 1.45 1.00 0.0145 Heptachlor 2 2.90 2.56 0.0743 2 2.90 0.50 0.0146 Parathion, ethyl 1 1.45 2.00 0.0290 3 4.35 1.40 0.0609 Vernolale 1 1.45 8.00 0.1159 83 TABLE 2 (cont'd). Pesticides applied to cropland by State, FY-70 No. Sites Using Pesticide Percent Sites Using Pesticide Arith. Mean Appl. Rate (LB/A)1 Akith. Mean (LB/A)1 Oklahoma, 56 Sites Borax 2 3.57 1.25 0.0446 Ceresan M 4 7.14 0.01 0.0007 Ceresan red 3 5.36 0.01 0.0005 2.4-D 2 3.57 0.75 0.0268 Disulfolon 1 1.79 0.50 0.0089 Parathion. ethyl 2 3.57 0.75 0.0268 Parathion, methyl 2 3.57 0.50 0.0179 PCNB 1 1.79 0.01 0.0002 Picloram 1 1.79 1.00 0.0179 Toxaphene 1 1.79 2.00 0.0357 Trifluralin 2 3.57 0.88 0.0313 Pennsylvania, 29 Sites Amitrole 1 3.45 2.11 0.0728 Airazine 4 13.79 2.16 0.2983 Azinphosmethyl 1 3.45 0.25 0.0086 2,4-D 3 10.34 0.10 0.0100 Malathion 2 6.90 1.13 0.0776 Methoxychlor 2 6.90 1.13 0.0776 Parathion, ethyl 1 3.45 0.25 0.0086 Silvex 1 3.45 0.81 0.0279 South Carolina, 9 Sites Carbaryl 3 33.33 0.37 0.1222 2.4-D 1 11.11 0.50 0.0556 DDT 1 11.11 2.00 0.2222 Mevinphos 2 22.22 0.10 0.0222 Parathion, ethyl 1 11.11 1.00 0.1111 TDE 1 11.11 3.00 0.3333 Toxaphene 2 22.22 0.80 0.1778 Trifluralin 3 33.33 0.60 0.2000 South Dakota, 106 Sites Aldrin 2 1.89 0.25 0.0048 Atra2ine 7 6.60 l.OO 0,0658 Barhan 1 0.94 0.20 0.0019 Bux-ten (bux) 1 0.94 10.00 0.0943 Captan 20 18.87 0.01 0.0020 Ceresan M 2 1.89 0.01 0.0002 2,4-D 26 24.53 0.48 0.1174 Dieldrin 2 1.89 0.01 0.0002 Heptachlor 1 0.94 0.06 0.0006 Lindane 1 0.94 0.01 0.0001 Malathion 15 14.15 on 0.0155 Maneb 1 0.94 0.01 0.0001 MCPA 2 1.89 0.17 0.0033 Methoxychlor 2 1.89 0.08 0.0015 Methylmercury dicyandiamide 7 6.60 0.01 0.0007 Thiram 2 1.89 0.01 0.0002 Toxaphene 1 0.94 2.00 0.0189 Tennessee, 23 Sites Atrazine 3 13.04 2.00 0.2609 Carbaryl 1 4.35 7.20 0.3130 Ceresan red 2 8.70 0.01 0.0009 Chloroxuron 2 8.70 1.13 0.0978 Cotoran 2 8.70 0.84 0.0730 2,4-D 4.35 1.50 0.0652 DAC 4.35 5.00 0.2174 Dalapon 4.35 4.00 0.1739 2,4-DB 8.70 0.19 0.0170 DDT 4.35 0.95 0.0413 Disulfoton 4.35 0.01 0.0004 EPTC 4.35 0.50 0.0217 Linuron 13.04 0.51 0.0670 MSMA 4.35 0.28 0.0122 Nitralin 4.35 0.60 0.0261 NPA 8.70 2.43 0.2113 Parathion, methyl 4.35 0.47 0.0204 PCNB 4.35 O.OI 0.0004 Sodium chlorate 4.35 4.50 0.1957 Toxaphene 4.35 1.90 0.0826 Trifluralin 4.35 1.11 0.0483 Virginia AND West Virginia, 21 Sites Atrazine 3 14.29 1.33 0.1905 Azinphosmethyl 1 4.76 0.50 0.0238 Captan 2 9.52 0.09 0.0086 DDT 1 4.76 0.25 0.0119 Lindane 1 4.76 0.01 0.0005 Malathion 1 4.76 0.01 0.0005 Sulan 1 4.76 3.00 0.1429 Toxaphene 1 4.76 0.50 0.0238 (Coruinued next page) 84 Pesticides Monitoring Journal TABLE 2 (cont'd). Pesticides applied to cropland by Slate, FY -70 Pesticide No. Sites Using Pesticide Pekcent Sites Using Pesticide Arith. Mean Appl. Rate (LB/A)1 Arith. Mean (LB/A)1 Wisconsin, 67 Sites Aldrin 1 1.49 5.00 0.0746 Amiben 2 2.99 11.50 0.3433 Atrazine 17 25.37 2.12 0.5373 Carbaryl 1.49 1.25 0.0187 2.4-D 2.99 0.57 0.0172 Diazinon 1 1.49 9.00 0.1343 Disulfoton 1.49 1.00 0.0149 MCPA 1.49 0.25 0.0037 Phorate 1.49 7.00 0.1045 Ramrod 1.49 1.50 0.0224 Trifluralin 1.49 1.50 0.0224 > To convert lb/a to kg/ha., multiply by 1.1208. 3 Use records were available for only 51 of the 150 sites in Illinois ^ Mid-Atlantic States include Maryland, Delaware, and New Jersey. ' New England States include Maine, New Hampshire, Vermont, Massachusetts, Rhode Island, and Connecticut. TABLE 3. Pesticides applied to cropland by crop, FY-70 No. Sites Using Pesticide Percent Sites Using Pesticide Arith. Mean Appl. Rate (LB/A)1 Arith. Mean (LB/A)1 Alfalfa and Bur Clover, 114 Sites Azinphosmethyl 2 1.75 0.38 0.0066 Carbaryl 1 0.88 1.00 0.0088 Malathion 2 1.75 1.13 0.0197 Methoxychlor 2 1.75 1.13 0.0197 Parathion, ethyl 3 2.63 0.50 0.0132 Field Corn, 366 Sites Aldrin 51 13.93 1.00 0.1398 Amiben 3 0.82 1.47 0.0120 Amitrole 1 0.27 2.11 0.0058 Atrazine 146 39.89 1.75 0.7001 Bux-ten (bux) 16 4.37 2.27 0.0990 Capian 87 23.77 0.02 0.0052 Carbaryl 5 1.37 1.49 0.0204 CDAA 1 0.27 0.60 0.0016 Ccresan red 1 0.27 0.10 0.0003 Chevron RE-5353 1 0.27 0.70 0.0019 2,4-D 72 19.67 0.64 0.1266 Dalapon 2 0.55 2.50 0.0137 2,4-DB 1 0.27 1.00 0.0027 DDT 1 0.27 0.25 0.0007 Diazinon 9 2.46 2.15 0.0528 Dicamba 1 0.27 0.02 0.0001 DIchlorprop 1 0.27 0.50 0.0014 Disulfoton 4 1.09 1.11 0.0121 Fensulfothion 4 1.09 1.20 0.0132 Furadan 1 0.27 1.00 0.0027 Heptachlor 20 3.46 0.79 0.0433 Isopestox 1 0.27 0.01 0.0000 Lasso 3 0.82 0.92 0.0076 Lindane 3 0.82 0.01 0.0001 Linuron 4 1.09 0.90 0.0098 Malathion 71 19.40 0.01 0.0019 Maneb 1 0.27 0.01 0.0000 MCPA 1 0.27 1.00 0.0027 Methoxychlor 18 4.92 0.02 O.OOU MSMA 1 0.27 3.00 0.0082 Nitrate 4 1.09 49.88 0.5451 Parathion, ethyl 1 0.27 0.02 o.onoi Phorate 16 4.37 1.23 0.0539 Ramrod 40 10.93 1.49 0.1628 Silvex 2 0.55 0.45 0.0023 Simazine 1 0.27 2.20 0.0060 Sutan 5 1.37 3.93 0.0540 Thiram 2 0.35 0.01 0.0001 Toxaphene 1 0.27 0.50 0.0014 {Continued next page) Vol. 8, No. 2, September 1974 85 TABLE 3 (cont'd). Pesticides applied to cropland by crop, FY-70 Pesticide No. Sites Using Pesticide Pehcent Sites Using Pesticide Arith. Mean Appl. Rate (LB/A)1 Arith. Mean (lb/a)i Cotton, 52 Sites Grass/Hay, 30 Sites Azinphosmethyl 1 1.92 3.00 0.0577 Azodrin 6 11.54 4.28 0.4942 Bidrin 8 15.38 0.31 0.0471 Calcium arsenate 1 1.92 9.80 0.1885 Captan 1 1.92 0.01 0.0002 Carbaryl 4 7.69 5.15 0.3962 Ceresan L 1 1.92 0.01 0.0002 Ceresan M 2 3.85 0.05 0.0021 Ceresan red 7 13.46 0.04 0.0048 Chlorobenzilate 1 1.92 1.00 0.0192 Chloroneb 5 9.62 0.01 0.0012 Cotoran 13 25.00 0.64 0.1608 DAC 1 1.92 4.00 0.0769 DDT 24 46.15 4.69 2.1625 DEF 6 11.54 1.44 0.1658 Dicofol 2 3.85 0.88 0.0337 Dimenthoate 1 1.92 0.33 0.0063 Dinitrocresol 2 3.85 0.10 0.0040 Disulfoton 4 7.69 0.01 0.0012 Diuron 8 15.38 0.47 0.0717 DSMA 6 11.54 2.02 0.2331 Endothall 1 1.92 0.90 0.0173 Endrin 3 5.77 4.16 0.2400 EPN 2 3.85 1.10 0.0423 Folex 7 13.46 1.30 0.1750 Linuron 2 3.85 0.79 0.0304 Malathion 1 1.92 2.50 0.0481 Methyl demeton 1 1.92 0.25 0.0048 Methylmercury dicyandiamide 1 1.92 0.01 0.0002 Methyl trithion 2 3.85 7.75 0.2981 Monuron 2 3.85 0.31 0.0121 MSMA 11 21.15 1.48 0.3138 Norea 1 1.92 1.00 0.0192 Nitralin 5 9.62 0.70 0.0669 Paraquat 6 11.54 0.18 0.0210 Parathion. ethyl 2 3.85 0.09 0.0035 Parathion, methyl 31 59.62 4.89 2.9129 PCNB 2 3.83 0.01 0.0004 Phorate 1 1.92 0.50 0.0096 Prometryne 1 1.92 0.75 0.0144 Sodium chlorate 4 7.69 6.88 0.5288 Strobane 1 1.92 3.00 0.0577 TDE I 1.92 6.00 0.1154 Toxaphene 20 38.46 6.94 2.6696 Trifiuralin 23 44.23 1.03 0.4550 Atrazine Simazine 3.33 3.33 2.50 2.50 0.0833 0.0833 Malathion Parathion, ethyl Mixed Hay, 119 Sites 0.84 0.84 1.00 2.00 0.0084 0.0168 Soybeans, 257 Sites Amiben 47 18.29 1.48 0.2710 Azinphosmethyl 1 0.39 4.00 0.0156 Captan 3 1.17 0.03 0.0004 Carbaryl 5 1.95 1.36 0.0265 CDEA 1 0.39 2.00 0.0078 Ceresan red 1 0.39 0.10 0.0004 CIPC 1 0.39 0.66 0.0026 Chloroxuron 3 1.17 1.42 0.0165 Cotoran 1 0.39 0.90 0.0035 DAC 1 0.39 5.00 0.0195 Dalapon 2 0.78 4.50 0.0350 2,4-DB 5 1.95 0.43 0.0083 DDT 3 1.17 16.65 0,1944 Dinitrobutylphenol 3 1.17 4.17 0.0487 Dinilrocyclohexylphenol 1 0.39 0.50 0.0019 EPTC 1 0.39 0.50 0.0019 Lasso 9 3.50 1.10 0.0386 Linuron 9 3.50 0.87 0.0303 Methylmercury dicyandiamide 1 0.39 0.01 0.0000 Nitralin 2 0.78 0.88 0.0068 NPA 6 2.33 2.02 0.0472 Parathion, ethyl 1 0.39 48.00 0.1868 Parathion. methyl 10 3.89 3.05 0.1186 Ramrod 3 1.17 1.33 0.0136 Toxaphene 3 1.17 49.30 0.5755 Trifiuralin 22 8.56 0.78 0.0670 Vernolate 2 0.78 4.65 0.0362 » To convert lb/a to kg/ha., multiply by 1.1208. 86 Pesticides Monitoring Journal TABLE 4. Chlorinated hydrocarbon residues in cropland soil, all sites^ — FY-70 Pesticide Aldrin Chlordane DAC o.p-DDE P,P'-DDE o,p'-DDT P.P'-DDT DDTR Dieldrin Endosiilfan (I) Endosulfan (II) Endosulfan sulfate Endrin Heptachlor Heptachlor epoxide Isodrin Lindane Nitralin Ramrod o.p -TDE P.p'-TDE Toxaphene Trifluralin No. Times 95% CONF. Range of Pesticide Percent Arith. Mean Geom. Mean Interval about Detected Detected Occurrence CONC, PPM CONC, PPM Geom. Mean, ppm Residues, ppm 203 13.48 0.02 0.0032 0.0027-0.0038 0.01- 4.25 165 10.96 0.08 0.0044 0.0036-0.0053 0.01- 13.34 1 0.07 <0.01 — — 1.19 43 2.86 <0.01 — — 0.01- 0.51 317 21.05 0.05 0.0062 0.0053-0.0071 0.01- 6.82 211 14.01 0.04 0.0037 0.0031-0.0043 0.01- 11.70 305 20.25 0.18 0.0085 0.0072-0.0098 0.01- 69.30 343 22.78 0.30 0.0116 0.0099-0.0134 0.01-113.09 465 30.88 0.04 0.0097 0.0086-0.0109 0.01- 1.85 1 0.07 <0.01 — — 0.01 4 0.27 <0.01 — — 0.02- 0.07 5 0.33 <0.01 — — 0.10- 0.29 27 1.79 <0.01 — — 0.01- 0.90 98 6.51 0.01 — — 0.01- 1.71 147 9.76 0.01 0.0016 0.0013-0.0019 0.01- 0.34 34 2.26 <0.01 — — 0.01- 0.18 6 0.40 <0.01 — — 0.01- 0.15 1 0.07 <0.01 — — 2.47 1 0.07 <0.01 — — 0.03 37 2.46 0.01 — — O.ni- 4.87 217 14.41 0.03 0.0033 0.0028-0.0038 0.01- 20.40 27 1.79 0.06 — — 0.79- 8.75 33 2.19 <0.01 — — 0.01- 0.36 NOTE: AH residues on dry-weight basis. ' 1,506 analyses, 35 States. TABLE 5. Chlorinated hydrocarbon residues in cropland soil by State, FY-70 No. Times Pesticide Detected Percent Occurrence Arith. Mean CONC, PPM Range of Detected Residues, ppm Chlordane DAC o,p'-DDE p,p'-DDE o.p'-DDT P.P-DDT DDTR Dieldrin Endrin Heptachlor epoxide Lindane o.p-TDE P.P-TDE Trifluralin Alabama, 21 Sites 1 1 2 17 13 16 17 4 2 1 2 2 12 6 4.76 4.76 9.52 80.95 61.90 76.19 80.95 19.05 9.52 4.76 9.52 9.52 57.14 28.57 0.01 0.06 <0.01 0.13 0.09 0.50 0.77 <0.01 0.01 <0.01 <0.01 0.01 0.04 0.02 0.23 1.19 0.02-0.08 0.02-0.49 0.01-0.47 0.03-3.08 0.05-3.83 0.01-0.03 0.05-0.16 0.03 0,01 0.04-0.08 0.01-0.20 0.02-0.13 Arkansas, 47 Sites Aldrin o,p'-DDE p.p'-DDE o.p-DDT p.p-DDT DDTR Dieldrin Endrin Lindane Nitralin o.p'-TDE P.P'-TDE Trifluralin 8 3 2S 15 22 25 12 3 1 1 2 18 2 17.02 6.38 53.19 31.91 46.81 53.19 25.53 6.38 2,13 2.13 4,26 38.30 4.26 <0.01 <0.01 0.13 0.06 0.47 0.69 0.03 <0.01 <0.01 0.05 0.01 0.03 0,01 0.01-0.03 0.01-0.03 0.01-1.25 0.01-0.62 0.03-3.98 0.02-5.30 0.04-0.22 0.01-0.03 0.01 2.47 0.09-0.21 0.01-0.31 0.01-0.36 California, 65 Sites Aldrin Chlordane o,p-DDE P,p'-DDE o.p-DDT PP-DDT DDTR Dieldrin Endosulfan (I) Endosulfan (II) Endosulfan sulfate Endrin Heptachlor expoxide o,p-TDE P.P'-TDE Toxaphene 1 2 14 41 32 40 46 23 1 2 2 5 2 12 34 13 1.54 3.08 21.54 63.08 49.23 61.54 70.77 35.38 1,54 3.08 3.08 7,69 3.08 18.46 52.31 20.00 <0.01 <0.01 0.01 0.17 0.08 0.38 0.74 0.05 <0.01 <0.01 0.01 <0.01 <0.01 0.01 0.08 0.61 0.17 0.10-0.20 0.01-0.38 0.01-1.39 0.01-0.62 0.01-5.16 0.01-7.75 0.01-1.28 0.01 0.04-0.06 0.17-0.29 0.02-0.10 0.01 0.01-0.26 0.01-1.20 0.79-7.63 (Continued next page) Vol. 8, No, 2, September 1974 87 TABLE 5 (cont'd). Clilorinated hydrocarbon residues in cropland soil by State, FY-70 Pesticide No. Times Pesticide Detected Percent Occurrence Arith. Mean CONC, PPM Range of Detected Residues, ppm Florida, 17 Sites Aldrin Chlordane P,p'-DDE o.p'-DDT P,P-DDT DDTR Dieldrin Endrin Heptachlor Heptachlor epoxide P.P-TDE Toxaphene 5.88 11.76 41.18 29.41 52.94 52.94 29.41 5.88 5.88 5.88 17.65 5.88 Georgia, 28 Sites <0.01 0.17 0.35 0.25 1.24 1.97 0.21 0.05 0.01 0.01 0.1.1 0.34 0.06 0.10- 2.72 0.01- 5.41 0.01- 4.18 0.02-20.26 0.03-31.99 0.05- 1.85 0.90 0.19 0.09 0.01- 2.14 5.71 o.p'-DDE p.p'-DDE o.p'-DDT P.P-DDT DDTR Lindane o.p-TDE P.P'-TDE Toxaphene Trifluralin 4 23 19 23 24 1 2 21 3 2 14.29 82.14 67.86 82.14 85.71 3.57 7.14 75.00 10.71 7.14 <0.01 0.22 0.13 0.74 1. 21 <0.01 <0.01 0.11 0.17 <0.01 0.01-0.02 O.Ol-i.09 0.02-0.84 0.02-3.38 0.03-5.11 0.07 0.02-0.09 0.01-0.99 1.21-2.27 0.02-0.03 Illinois, 140 Sites Aldrin Chlordane P.p'-DDE o.p-DDT P.P-DDT DDTR Dieldrin Heptachlor Heptachlor epoxide Isodrin Lindane PP-TDE Trifluralin 73 48 11 2 13 14 96 45 46 20 1 5 1 52.14 34.29 7.86 1.43 9.29 10.00 68.57 32.14 32.86 14.29 0.71 3.57 0.71 0.08 0.25 0.01 <0.01 0.01 0.01 0.15 0.03 0.02 <0.01 <0.01 <0.01 <0.01 0.01-1.38 0.05-3.76 0.02-0.16 0.01-0.02 0.02-0.20 0.03-0.33 0.02-0.92 0.01-0.65 0.01-0.34 0.01-0.04 0.02 0.02-0.04 0.03 Indiana, 78 Sites Aldrin 15 19.23 0.07 0.01-1.61 Chlordane 7.69 0.07 0.09-1.51 Dieldrin 17 21.79 0.04 0.02-0.58 Heptachlor 8.97 0.01 0,01-0.21 Heptachlor epoxide 6.41 <0.01 0.01-0.12 Isodrin 6.41 <0.01 0.01-0.18 Trifluralin 1.28 <0.01 0.06 Iowa, 150 Sites Aldrin Chlordane P.P'-DDE o.p'-DDT P.P-DDT DDTR Dieldrin Heptachlor Heptachlor epoxide Isodrin Lindane Ramrod P.P'-TDE 43 30 10 6 14 14 95 19 33 1 1 1 8 28.67 20.00 6.67 4.00 9.33 9.33 63.33 12.67 22.00 0.67 0.67 0.67 3.33 0.03 0.17 <0.01 <0.01 0.01 0.02 0.07 0.02 0.01 <0.0! <0.01 <0.01 <0.01 0.01-0.68 0.01-8.04 0.01-0.22 0.01-0.09 0.02-0.4 i 0.03-0.72 0.01-0.56 0.01-1.71 0.01-0.28 0.01 0.15 0.03 0.01-0.05 Kentucky. 30 Sites Aldrin o.p'-DDE P.p'-DDE o.p'-DDT P.P-DDT DDTR Dieldrin o.p-TDE P.P'-TDE Toxaphene (Continued next page) 3.33 3.33 26.67 10.00 20.00 26.67 26.67 3.33 20.00 3.33 <0.01 <0.01 0.01 0.01 0.02 0.04 0.01 <0.01 0.01 0.03 0.04 0.01 0.01-0.15 0.02-0.09 0.02-0.31 0.02-0.59 0.01-0.06 0.03 0.01-0.11 0.89 88 Pesticides Monitoring Journal TABLE 5 (cont'd). Chlorinated hydrocarbon residues in cropland soil by State, FY -70 Pesticide No. Times Pesticide Detected Percent Occurrence Arith. Mean CONC, PPM Range op Detected Residues, ppm Louisiana, 26 Sites Aldrin 1 3.85 <0.01 0.04 o,p'-DDE 2 7.69 <0.01 0.02-0.10 p,p'-DDE 12 46.15 0.20 0.02-1.98 o,p'-DDT 10 38.46 0.07 0.01-0.56 p.P -DDT 12 46.15 0.40 0.06-3.21 DDTR 12 46.15 0.70 0.09-6.02 Dieldrin 8 30.77 0.02 0.02-0.08 Endrin 2 7.69 <0.01 0.02-0.03 Heptachlor 1 3.85 <0.01 0.01 p,p'-TDE 11 42.31 0.03 0.01-0.17 Toxaphene 1 3.85 0.20 5.32 Trifluralin 2 7.69 0.01 0.07-0.19 Michigan, 54 Sites Chlordane 5.56 0.01 0.07- 0.51 o,p'-DDE 1.85 0.01 0.51 p,p'-DDE 12.96 0.06 0.01- 2.38 o,p'-DDT 9.26 0.16 0.04- 8.21 p.p-DDT 11.11 0.26 0.03-12.03 DDTR 12.96 0.53 0.01-25.59 Dieldrin 12.96 0.01 0.01- 0.22 Endosulfan (ID 1.85 <0.0I 0.02 Endosulfan sulfate 1.85 <0.01 0.10 Heptachlor 1.85 <0.01 0.04 Heptachlor epoxide 2 3.70 <0.01 0.01 o.p-TDE 1 1.85 0.02 1.30 p.p-TDE 5 9.26 0.03 0.02- 1.16 Mid-Atlantic States.' 19 Sues Aldrin 1 5.26 0.01 0.24 Chlordane 4 21.05 0.72 0.11-13.34 p.p-DDE 3 15.79 0.02 0.01- 0.22 o.p -DDT 2 10.53 <0.01 0.02- 0.03 p,p-DDT 5 26.32 0.03 0.01- 0.38 DDTR 5 26.32 0.10 0.01- 1.46 Dieldrin 9 47.37 0.08 0.01- 0.74 Heptachlor epoxide 1 5.26 <0.01 0.03 p.P-TDE 3 15.79 0.05 0.02- 0.86 Minnesota, 120 Sites Aldrin 6 5.00 0.05 0.03-4.25 Chlordane 8 6.67 0.02 0.03-0.94 o,p'-DDE 1 0.83 <0.01 0.01 p,p-DDE 12 10.00 0.01 0.01-0.25 o,p'-DDT 9 7.50 <0.01 0.01-0.25 p.P-DDT 13 10.83 0.02 0.02-0.74 DDTR 13 10.83 0.03 0.02-1 .29 Dieldrin 17 14.17 0.02 0.01-0.92 Endrin 2 1.67 <0.01 0.01-0.02 Heptachlor 3 2.50 <0.01 0.01-0.05 Heptachlor epoxide 5 4.17 <0.01 0.01-0.09 Isodrin 2 1.67 <0.01 0.01-0.05 P.p-TDE 4 3.33 <0.0I 0.01-0.08 Mississippi, 29 Sites Chlordane 1 3.45 <0.01 0.08 o,p-DDE 8 27.59 0.01 0.02-0.05 p,p'-DDE 27 93.10 0.28 0.01-1.71 o.p-DDT 22 75.86 0.19 0.02-0.74 P.P -DDT 27 93.10 1.29 0.01-6.61 DDTR 27 93.10 1.85 0.03-9.81 Dieldrin 6 20.69 0.02 0.01-0.18 Endrin 3 10.34 0.01 0.02-0.11 Heptachlor epoxide 1 3.45 <0.01 0.03 P,P-TDE 18 62.07 0.08 0.01-0.7! Toxaphene 5 17.24 0.73 2.79-8.75 Trifluralin 5 17.24 0.03 0.03-0.27 Missouri, 81 Sites Aldrin 20 24.69 0.04 0.01-0.49 Chlordane 9 11.11 0.16 0.07-5.62 P.P-DDE 6 7.41 <0.01 0.01-0.10 o.p -DDT 2 2.47 <0.01 0.03-0.04 p.P -DDT 6 7.41 0.01 0.03-0.26 DDTR 6 7.41 0.01 0.04-0.43 Dieldrin 28 34.57 0.06 0.01-0.53 Endrin 1 1.23 <0.01 0.02 Heptachlor 8 9.88 0.03 0.02-0.94 Heptachlor Epoxide 8 9.88 0.01 0.01-0.31 Isodrin 1 1.23 <0.01 0.01 p,p-TDE 3 3.70 <0.01 0.02-0.04 Trifluralin 5 6.17 <0.01 0.02-0.09 (Continued next page) Vol. 8, No. 2, September 1974 89 TABLE 5 (cont'd). Chlorinated hydrocarbon residues in cropland soil by Slate, FY-70 Pesticide No. Times Pesticide Detected Percent Occurrence Arith. Mean Cone, PPM Aldrin Chlordane o.p'-DDE P.p-DDE o.p-DDT P.P-DDT DDTR Dieldrin Endrin Heptachlor epoxide p,p-TDE Chlordane o,p-DDE P.P-DDE o,p'-DDT P,p'-DDT DDTR Dieldrin Endosulfan (II) Endosulfan sulfate P,p'-TDE Nebraska, 106 Sites 4 16 1 10 5 7 10 48 4 18 3 3.77 15.09 0.94 9.43 4.72 6.60 9.43 45.28 3.77 16.98 2.83 <0.01 0.01 <0.01 <0.01 <0.01 0.01 0.01 0.03 <0.01 <0.01 <0.01 New England States,^ 20 Sites Range of Detected Residues, ppm 0.01 0.02-0.14 0.01 0.01-0.19 0.01-0.06 0.03-0.28 0.01-0..S9 0.01-0.17 0.01-0.03 0.01-0.02 0.01-0.03 5.00 5.00 25.00 25.00 25.00 25.00 10.00 5.00 5.00 20.00 0.01 <0.01 0.07 0.07 0.45 0.65 0.03 <0.01 0.01 0.05 0.19 0.03 0.01-0.68 0.01-0.86 0.02-4.63 0,04-6.65 0.32-0.33 0.07 0.23 0.14-0.43 New York, 38 Sues Chlordane P.P-DDE o.p-DDT P.P-DDT DDTR Dieldrin Heptachlor epoxide o.p'-TDE P,p'-TDE 1 13 6 12 15 10 1 1 6 2.63 34.21 15.79 31.58 39.47 26.32 2.63 2.63 15.79 0.01 0.21 0.31 1.87 3.06 0.01 <0.01 0.13 0.54 0.28 0.01- 6.82 0.01- 11.70 0.01- 69.30 0.01-113.09 0.01- 0.10 0.06 4.87 0.01- 20.40 Aldrin Chlordane o,p-DDE P.P-DDE o.p'-DDT P.P-DDT DDTR Dieldrin Endrin Heptachlor epoxide o,p-TDE P.p-TDE Trifluralin North Carolina, 30 Sites 1 3 I 21 16 20 22 10 2 1 8 18 1 3.33 10.00 3.33 70.00 53.33 66.67 73.33 33.33 6.67 3.33 26.67 60.00 3.33 <0.01 0.01 <0.01 0.07 0.06 0.31 0.53 0.03 <0.01 <0.01 0.01 0.07 <0.01 0.02 0.03-0.32 0.02 0.01-0,44 0.02-0.43 0.02-2.93 0.03-4.12 0.01-0,29 0.01-0,03 0,01 0,01-0,12 0.01-0.42 0.07 Ohio, 69 Sites Aldrin Chlordane P,P-DDE o.p'-DDT P.P-DDT DDTR Dieldrin Endrin Heptachlor Heptachlor epoxide Isodrin o.p-TDE P.P-TDE Trifluralin 17 15 6 1 3 7 26 1 6 9 2 4 6 2 24.64 21.74 8.70 1.45 4.35 10.14 37.68 1.45 8.70 13.04 2.90 3.80 8.70 2.90 0.04 0.08 <0.01 <0.01 ,P'-DDE 33 12.22 <0.01 0.01-0.36 .'.P-DDT 33 12.22 0.0! 0.01-0.28 P.P'-DDT 53 19.63 0.03 0.01-2.20 DDTR 54 20.00 0.04 0.01-2,88 Dieldrin 42 15.56 <0.01 0.01-0.09 Heplachlor 1 0.37 <0.01 0.06 Hepiachlor epoxide 5 1.85 <0.01 0.01-0.12 P,P-TDE 29 10.74 <0.01 0.01-0.30 Toxaphene 12 4,44 0 02 0,09-1 35 Orcanophosphates, 22S Siii'i Malathion 1 1 0.44 <0.01 0.04 {Continued next page) 96 Pesticides Monitoring .Iournal TABLE 11 (cont'd). Chlorinated hydrocarbon and organophosphale residues in crops, all sites, FY -70 No. Times Pesticide Detected p,p-DDE o,p-DDT p.p-DDT DDTR Dieldrin p,P-TDE Toxaphene Percent Occurrence Arith. Mean CONC, PPM Cotton: Seeds Chlorinated Hydrocarbons, 39 Sites Range of Detected Residues, ppm 13 14 20 20 2 8 12 33.33 35.90 51.28 51.28 5.13 20.51 30.77 2.57 0.01 0.07 2.65 <0.01 <0.01 0.15 O.OI-lOO.Ol O.OI- 0.18 0.01- 1.02 0.01-100.03 0.01 0.01- 0.02 0.05- 1.85 Organophosphates, 37 Sites DEF Parathion, ethyl Parathion, methyl 17 3 7 45.95 8.11 18.92 0.11 <0.01 0.01 0.02-1.10 0.01-0.03 0.01-0.08 Mixed Hay Chlorinated Hydrocarbons, 29 Sites Aldrin 1 3,45 <0.01 0.07 Chlordanc 5 17.24 0.07 0.05-1.19 P.p-DDE 11 37.93 0.01 0.01-0.19 o,p-DDT 11 37.93 0.01 0.01-0.04 P,P-DDT 20 68.97 0.02 0.01-0.13 DDTR 20 68.97 0.04 0.01-0.36 Dieldrin 19 65.52 0.01 0.01-0.11 Endrin 1 3.45 <0.01 0.12 Heptachlor 1 3.45 <0.01 0.02 P,P-TDE 6 20.69 <0.01 0.01-0.02 Toxaphene 4 13.79 0.01 0.05-0.13 Organophosphates, 29 Sites; 0 Detections Soybeans: Beans Chlorinated Hydrocarbons, 178 Sites Chlordanc 3 1.69 <0.01 0.01-0.10 Dieldrin 69 38.76 0.01 0.01-0.09 Endrin 15 8.43 <0.01 0.01-0.14 Heptachlor 2 1.12 <0.01 0.01 Heptachlor epoxide 12 6.74 <0.01 0.01-0.05 Ramrod 1 0.56 <0.01 0.24 Toxaphene 19 10.67 0.02 0.08-0.41 Trifluralin 8 4.49 <0.01 0.01-0.09 Organophosphates, 137 Sires: 0 Detections NOTE: Samples were examined for organophosphates only when use records indicated they had been applied. All residues on dry-weight basis. Vol. 8, No. 2, September 1974 97 DDT Moratorium in Arizona — Agricultural Residues after 4 Years'^ G. W. Ware, B. J, Estesen, and W. P. Cahill ABSTRACT The moratorium on agriciilturcit use of DDT in Arizona thai began in January 1969 has proved very effective during the 4 years of its enforcement. Residues on green alfalfa have declined significantly during this period to a probable in- herent level of 0.03 ppm, fvet weight. Soil residues of DDTR have changed almost imperceptibly: this suggests a soil half- life greater than 20 years. These soil residues are mostly DDE; the little remaining DDT is being converted gradually to DDE, the slowly degraded metabolite of DDT. This paper retracts a statement made in an earlier study which implied violation of the DDT moratorium in the Yuma mesa and valley in Yuma County. Authors have con- cluded thai the con.'sistenlly high residues in the Yuma area resulted from climatological conditions not found in other sampling areas, rather than from an\ violation of the mora- torium. Iiilrodiiction The current moratorium on agricultural use of DDT in Arizona that began in January 1969 has completed its fourth year (1.2). This is the third report on the status of DDT residues and DDTR. related degradation products, following 1 8 years of unrestricted use and 4 years of restricted use in Arizona under the guidance of the Arizona Board of Pesticide Control. A nalylical Methods Soil and alfalfa samples were collected precisely as described in previous reports (1 .2) from the three major irrigated areas in Arizona: Salt River Valley, which Contribution to Regional Project W-45, "Residues of Pesticides and Related Chemicals in the Agricultural Environment — Their Nature, Distribution. Persistence, and Toxicological Implications." University of Arizona Agricultural Experiment Station Journal Series No. 2139. Department of Entomology, University of Arizona. Tucson, Ariz. 85721. surrounds Phoenix; Pinal County; and the Yuma mesa and valley in Yuma County. Desert soil samples adja- cent to these areas were also collected but only from the top 0.25 inch. Authors had hoped to demonstrate that airborne contaminated dust was a prime source of forage contamination. This theory was disproved. Addi- tionally, an earlier study (3) was continued to provide reference standards and residue continuity retrospec- tively through 1967 (Table 1). The sampling sites are located on a 60-mile Maricopa County east-west transect along Baseline Road. Alfalfa and soil samples were carried through the extraction and cleanup procedures formerly delineated by the authors {1-3}. Analysis was performed by electron-capture/gas-liquid chromatography (EC/GLC). Recovery standards and analytical reagent blanks were carried through the extraction and cleanup procedures for each day's analy- sis. Recoveries were consistently above 90 percent; how- ever, corrections were not applied to the data presented. The minimum sensitivity of the method was arbitrarily set at 0.02 ng for p.p'- and o.p'-DDT. DDE, and DDD. Standard curves extended from 0.03 to 0.10 ng. The relative sensitivities were 0.001 ppm for alfalfa and 0.003 ppm for soil. Results are based on a minimum sample size and 6 \i\ extract injected into the chroma- tograph. Analytical EC/GLC confirmatory tests were conducted on a random basis using a double-length GLC column at a temperature slightly higher than that used in pre- vious studies, as well as p-value determinations using acetonitrile and hexane (4). Because of low levels of DDTR and interfering peaks of toxaphene which drifted from nearby cottonfields. all alfalfa extracts were dehy- drohalogenated after cleanup on fiorisil and measured only as o,p'- and p,p'-DDE as described by Cahill et al. (5). 98 Pesticides Monitoring Journal TABLE 1. DDTR residues ippm] in green alfalfa. 1967-72, Maricopa County, Ariz. 1967 1968 1969 1970 1971 1972 Sample Aug. Sept. Sept. Sept. Sept. Sept. 2 0.220 0.038 0.050 0.020 0.023' 3 0.283 — 0.027 0.030 — 0.025* 4 0.170 0.120 0.038 0.037 0.031 0.022 5 — 0.060 0.020 0.024 0.011 0.029* 6 0.277 — 0.035 0.022 — 0.008* 8 0.794 — — 0.027 0.038 0.013* 9 — 0.076 0.034 0.042 0.020 0.029* 10 0.350 0.092 0.054 0.162 0.027 0.031 11 0.453 0.580 0.064 0.047 0.085 0.056 12 0.299 0.077 0.025 0.038 — 0.023* 13 0.606 — — 0.021 0.027 — Means' 0.404' 0.175" 0.037« 0.045» 0.032" 0.026» NOTE: — = no samples analyzed; * — substitute adjacent fields. ' Means with same letter are not significantly different at the 0.05 level. TABLE 2. DDTR residues (ppm) in green alfalfa during 1969-7} DDT moratorium, Maricopa County, Ariz. 1969 1969 1970 1971 1972 Sample Jan. Sept. Sept. Sept. Sept. 1 0.087 0.042 0.057 2 0.303 0.062 0.050 0.025 0.039* 3 0.102 0.078 0.093 0.038 — 4 0.107 0.047 0.076 0.037 0.046* 5 0.049 0.030 0.025 0.007 0.01 1 6 0.113 0.064 0.060 0.051 0.045* 7 0.082 0.034 0.023 — 0.055 8 0.125 0.056 — — — 9 0.085 0.044 0.101 — — 10 — — 0.080 0.059 — Mean;;' o.in" 0.051« 0.063- 0.036' 0.039« NOTE: — = no samples analyzed; * = substitute adjacent fields. 1 Means with same letter are not significantly different at the 0.05 level. TABLE 3. DDTR residues (ppm) in green alfalfa during 1969-71 DDT moratorium, Pinal County, Ariz. Sample 1969 Jan. 0.047 0.047 0.142 0.231 0.092 0.038 0.079 0.068 0.054 0.088" 1969 Sept. 0.042 0.031 0.187 0.076 0.130 0.058 0.118 0.071 0.068 0.086>' 1970 Sept. 0.034 0.059 0.071 0.045 0.045 0.059 0.031 0.057 0.050" 1971 Sept. 0.055 0.036 0.072 0.038 0.0.34 0.060 0.049" 1972 Sept. 0.041* 0.025 0.025* 0.044 0.018 0.031* NOTE: — = no samples analyzed; * = substitute adjacent fields. ' Means with same letter are not significantly different at the 0.05 level. TABLE 4. DDTR residues (ppm) in green alfalfa during 1969-71 DDT moratorium, Yuma County, Ariz. 1969 1969 1970 1971 1972 1972 Sample Jan. Sept. Sept Sept. Jan. Sept. 1 0.047 0.373 0.120 0.025 0.032 2 0.039 0.098 — — 0.010* 0.017* 3 0.049 0.256 0.084 0.270 0.073* 0.040* 4 0.057 0.093 — — 0.055* 0.075* 5 0.057 0.545 0.063 0.340 0.047* 0.290* 6 0.044 0.317 — — 0.035* 0.300* 7 0.059 0.241 — — 0.026* 0.190* 8 0.036 0.045 0.034 0.031 0.039* — 9 0.021 0.056 — — 0.015* — 10 0.046 0.074 0.051 0.050 0.028 0.045 Means' 0.046* 0,210"' 0.058" 0.162i> 0.035" 0.123i> NOTE: — = no samples analyzed; * = substitute adjacent fields. ' Means with same letter are not significantly different at the 0.05 level. TABLE 5. DDTR residues {ppm) in soils during 1969-72 moratorium, Maricopa County, Ariz 1969 1970 1972 Field No. Jan. Sept. Sept. DDE o.p'- P,P'- Total' DDE o.p'- P.P'- Total DDE O.p'- P,p'- Total DDT DDT DDT DDT DDT DDT 1 0.35 0.04 0.12 0.54 0.32 0.04 0.11 0.47 0.40 0.04 0.11 0.55 2 0.48 0.17 0.78 1.54 0.79 0.20 0.96 1.95 0.98 0,18 0.47 1.63 3 0.33 0.07 0.16 0.59 1,35 0.13 0.32 1.80 1,24 0,13 0.32 1.69 4 0.49 0.05 0.17 0,74 0,56 0.06 0.22 0.84 0,58 0,05 0.23 0.86 5 0.29 0.05 0.09 0,44 0.15 0,02 0.05 0,22 0.17 0,01 0.05 0.23 6 2.10 0.43 1.10 3,93 2.57 0.39 1,18 4,14 2.58 0,28 0.96 3.82 7 0.84 O.Il 0,23 1,22 0,84 0,13 0,23 1.20 0.92 0,09 0.29 1.30 8 2.22 0.3S 1,29 4,00 2,50 0,47 1,61 4.58 2.37 0,27 1.21 3.85 9 1.18 0.21 0,91 2,41 1,24 0,24 0.82 2.30 1.12 0.17 0.77 2.06 10 — — — (0.24) 0.48 0.06 0.14 0.68 0.31 0.04 0.07 0.42 Means2 0.92 0.17 0.54 1.57" 1.08 0.17 0.56 1.82"'' 1.07 0.13 0.45 1.64* Desert Sample 1 0.08 <0.01 0.03 0.13 0.48 0.04 0.09 0.61 0.43 0.07 0.09 0.59 2 0.24 0.02 0,06 0.35 0.41 0.05 0.10 0.56 0.28 0.03 0.58 0.89 3 0.44 0.04 0.15 0.67 0.28 0.02 0.07 0.37 0.18 0.02 0.04 0.24 4 — — — (2.39) 1.44 0.09 0.38 1.91 0.54 0.08 0.06 0.68 Means= — — — 0.89" 0.65 0.05 0.16 0.86* 0.36 0.05 0.19 0.60* NOTE: — = no samples analyzed, ' Figures in parentheses are missing values calculated by randomized blocks missing value formula. ■ Means with same letter are not significantly different at the 0.05 level. Vol. 8, No. 2, September 1974 99 TABLE 6. DDTR residues (ppm) in soils during 1969-72 moratorium, Pinal County, Ariz. 1969 1970 1972 Field Jan. Sei-t. Sept. No. DDE o,p'- DDT P.p'- DDT Total DDE o,p'- DDT P.P'- DDT Total' DDE o.p'- DDT P.P'- DDT Total 1 0.64 0.48 2.43 3.77 0.61 0.35 2.82 3.78 0.74 0.34 2.64 3.72 2 0.27 0.15 1.03 1.52 0.35 0.11 1.00 1.46 0.41 0.13 0.96 1.50 3 1.05 0.32 1.38 2.75 1.32 0.23 1.03 2.58 1.16 0.16 0.80 2.12 4 0.99 0.27 1.04 2.30 1.23 0.24 0.68 2.15 1.40 0.18 0.74 2.32 5 0.16 0.02 0.21 0.41 — — — (0.38) 0.25 0.02 0.16 0.43 6 0.06 0.01 0.07 0.14 — _ — (0.06) 0.07 0.01 0.04 0.12 7 1.09 0.28 1.37 2.74 1.41 0.31 1.30 3.02 1.63 0.20 0.80 2.63 8 0.09 <0.01 0.04 0.14 0.06 0.01 0.02 0.09 0.08 0.01 0.02 0.11 9 0.67 0.09 0.29 1.06 0.79 0.08 0.20 1.07 0.74 0.03 0.06 0.83 10 0.66 0.14 0.36 1.16 1.14 0.12 0.27 1.53 1.19 0.15 0.39 1.73 Means" 0.57 0.18 0.82 1.60" 0.72 0.15 0.76 1.61« 0.69 0.12 0.66 1.55' Desert Sample 1 0.09 <0.01 0.06 0.16 0.18 0.01 0.03 0.22 0.17 0.02 0.12 0.31 2 0.18 0.01 0.11 0.32 0.38 0.03 0.06 0.47 0.21 0.02 0.21 0.44 3 0.05 0.03 0.10 0.21 0.12 0.02 0.05 0.19 0.06 0.01 0.02 0.09 4 0.09 0.03 0.10 0.25 1.18 0.05 0.16 1.40 0.77 0.07 0.09 0.93 Means= 0.10 0.02 0.09 0.24- 0.47 0.03 0.08 0.57" 0.30 0.03 0.11 0.44» NOTE: — = no samples analyzed. ^ Figures in parentheses are missing values calculated by randomized blocks missing value formula. 2 Means with same letter are not significantly different at the 0.05 level. TABLE 7. DDTR residues (ppm) in soils during 1969-72 moratorium, Yuma County, Ariz- 1969 1970 1972 Field Jan. Sept. Sept. No. DDE O.p'- DDT P.p'- DDT Total DDE O.p'- DDT P.p'- DDT Total DDE o,p'- DDT P,P'- DDT Total 0.10 <0.01 0.07 0.17 0.11 0.02 0.06 0.19 0.12 0.02 0.03 0.17 0.24 0.05 0.25 0.54 0.35 0.10 0.25 0.70 0.20 0.03 0.07 0.30 0.72 0.16 0.72 1.60 0.66 0.18 0.52 1.36 0.79 0.16 0.49 1.44 0.59 0.11 0.47 1.17 0.78 0.17 0.49 1.44 0.98 0.15 0.46 1.59 0.48 0.05 0.30 0.83 0.44 0.12 0.31 0.87 0.75 0.12 0.34 1.21 6 0.29 0.16 0.74 1.19 0.40 0.14 0.56 1.10 0.48 0.10 0.43 1.01 7 1.29 0.07 0.37 1.73 1.09 0.11 0.35 1.55 1.11 0.09 0.60 1.80 8 0.06 0.01 0.01 0.08 0.00 0.00 0.00 0.04 0.05 <0.01 <0.01 0.07 9 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 <0.01 <0.01 <0.0I 0.03 10 0.26 0.02 0.03 0.31 0.22 0.02 0.03 0.27 0.17 0.01 0.02 0.20 Meansi 0.40 0.06 0.30 0.76« 0.41 0.09 0.26 0.75« 0.47 0.07 0.25 0.78" Desert Sample 1 0.27 0.02 0.07 0.36 0.22 0.03 0.09 0.34 0.24 0.06 0.09 0.39 2 0.03 0.01 0.02 0.06 0.08 0.01 0.04 0.13 0.02 0.01 0.02 0.05 3 0.02 0.01 0.03 0.06 0.02 0.00 0.03 0.05 0.02 0.01 0.02 0.05 4 0.00 0.00 0.01 0.01 0.00 0.00 0.01 0.01 0.79 0.07 0.15 1.04 Means^ 0.08 0.01 0.03 0.1 2» 0.08 0.01 0.04 0.14" 0.27 0.04 0.08 0.38" 1 Means with same letter are not significantly different at the 0.05 level. Results and Discussion Residues observed in alfalfa and soil samplings during the past 4 years are presented in Tables 1-7 as DDTR. The Student-Newman Keul's test was used to analyze differences among residue means for the various sam- pling dates. Comparisons were made on least-squares means in the soil samples (Tables 5-7) due to inade- quate samples. Residues on alfalfa from all four areas shown in Tables 1-4 appear to have leveled off at about 0.03 ppm except in Yuma County (Table 4), where September residues from 1969 through 1972 were about threefold higher than were September residues from other areas. Sampling in January 1972 indicated that residues in Yuma had declined to the levels of the other areas. High September values for Yuma have occurred consistently in 1969, 1970, and 1971. This condition requires the retraction of a statement in the last report (2) indicating violations of the DDT mora- torium in the Yuma area. These phenomenally high residues are apparently the result of climatological con- ditions not found in the other sampling areas. Residue levels in alfalfa soils did not appear to decline from the last sampling period. September 1970 (Tables 5-7). Because any decline has been imperceptible, the 100 Pesticides Monitoring Journal time required for these residues to reach one-half their present level is now estimated to be greater than 20 years, with desert soils changing the least. DDTR residues now found in Arizona alfalfa and soil are primarily DDE. DDT residues are slowly becoming DDE and then declining negligibly. As suggested from past studies, future problems arising from DDT will be attributable to DDE, the vpry slowly degrated metabolite of DDT. LITERATURE CITED (7) Ware, G. W., Betty Estesen, C. D. John, and W. P. Cahill. 1970. DDT moratorium in Arizona — agricul- tural residues after 1 year. Pestic. Monil. J. 4(1) :21-24. (2) Ware, G. W., Betty Estesen, and W. P. Cahill. 1971. DDT moratorium in Arizona — agricultural residues after 2 years. Peslic. Monit. J. 5(3 ) :276-280. (3) Ware, G. W.. Betty Estesen, and W. P. Cahill. 1968. Pesticides in soil — an ecological study of DDT residues in Arizona soils and alfalfa. Pestic. Monit. J. 2(3): 129-132. (4) Bowman, M. C, and Morton Beroza. 1965. Extraction p-values of pesticides and related compounds in six binary solvent systems. J. Assoc. Off. Agric. Chem. 48(5):943-952. (5) Cahill, W. P., Betty Estesen, and G. W. Ware. 1970. Determination of DDT in the presence of toxaphene residues. Bull. Environ. Contam. Toxicol. 5(3) : 260-262. Vol. 8, No. 2, September 1974 101 RESIDUES IN FISH, WILDLIFE, AND ESTUARIES Mercury Residues in the Common Pigeon (Columba livia) from the Jackson, Mississippi, Area — 7972 Luther A. Knight, Jr.,' and Edward J. Harvey, Sr.' ABSTRACT Total mercury in the common pigeon (Columba livia) from the Jackson, Miss., area was measured by atomic absorption spectrophotometry. Pigeons captured in 1972 from down- town Jackson were killed on the day of capture to determine mercury levels in wild birds in urban environments; others were caged and analyzed for mercury residues weekly or biweekly for 9 weeks. Data are presented to show possible pathways by which organisms eliminate this element. Median concentration of mercury in brains of newly captured pigeons was 22 ppb. Claws showed 14 to 85 ppb mercury. Possible sources of mercury contamination in these birds are treated grains, contaminated weed seeds, and naturally occurring mercurials. Introduction Even at trace levels, certain contaminants may have effects on the ecosystem as great or greater than those of the more common pollutants (1). Mercurials have been used for many years in agriculture, industry, and medi- cine, and have caused much concern about their effects on the biosphere. In his review of mercury uses in Canada. Fimreite emphasized that mercury contamina- tion has been investigated in Scandinavian countries where it has produced greater wildlife problems than have DDT and other organochlorine pesticides used prevalently in Scandinavia (2). The seriousness of water pollution by mercury in industrial effluents was accen- tuated by the occurrence of Minamata disease in Japan (3) and. as a consequence of the potential human health hazard, considerable emphasis has been placed upon the danger of mercurials in the aquatic environment in the United States and Canada. After reviewing sources and uses of mercury. Summer. Saha. and Lee stated that elevated levels of the element may be expected in ^ Department of Biology, University of Mississippi, University, Miss. 38677. ' Department of Biology, Jackson State University, Jackson, Miss. 39217. fish from waters receiving mercury containing industrial: and/or municipal wastes (4). In particular, fish have been used to study the extenti and effects of this type of pollution. A number of States have banned the sale of commercial fish or warned against public consumption of fish from contaminated waters (5). Fimreite, Fyfe. and Keith (6) demonstrated widespread contamination among seed-eating birds in areas of Canada where mercury-containing fungicides are used in seed treatment. A decline in seed-eating bird populations was traced to the use of mercurials as fungicides in Sweden (7). Findings of elevated levels of mercury in various game birds have prompted several States and the Canadian provinces of Alberta and Sas- katchewan to monitor mercurial concentrations in their wildlife (2,6,8,9). In Mississippi, Knight and Herring reported concen- trations of total mercury from less than 0.05 to 0.74 ppm in muscles of largemouth bass from Ross Barnett Reservoir (10). However, there is a lack of data on the extent of trace metal contamination in ecosystems of Mississippi. Organisms which accumulate trace metals can act as indicators of pollution levels in their environ- ments. In view of the paucity of local data on trace elements, a study was initiated to evaluate possible indi- cator organisms and to measure mercury and lead in the Jackson. Miss., area. This paper reports the pre- liminary results of monitoring total mercury in the common pigeon (Columba livia). A nalytical Methods Wild pigeons were chosen as test animals because of their abundance and adaptation to urban environments. Approximately 100 pigeons were captured in an aban- doned hotel in downtown Jackson on October 10. 1972. Researchers killed 25 of the birds that day and placed the remainder in outside, off-the-ground cages 102 Pesticides Monitoring Journal for analysis on a weekly schedule. Birds were fed a commercial pigeon scratch and corn which contained no detectable amounts of mercury residues. This control diet assured authors that any mercury detected in pigeons during the 9 weeks of analyses originated from their urban eating patterns before capture rather than from their control diets. The flameless atomic absorption method described by Hatch and Ott (//) was used to measure total mercury in all birds: those killed at time of capture and those killed during the next 9 weeks. A 1- to 3-g sample of tissue was digested with concentrated nitric and sulfuric acids in a 125-ml flask at 50^C to destroy the organic matter. After dissolution, samples were cooled to room temperature and 6 percent potassium permanganate was added until a faint color persisted. To eliminate excess oxidants, 30 ml of reductant containing hydroxylamine sulfate was added; stannous sulfate was used to reduce mercury compounds to the elemental state. The flask was then placed in the absorption system and removed after maximimi absorption was reached. Mercury vapor was vented to the hood. Standards obtained from Beck- man Instruments. Inc., Fullerton, Calif., were run in the same manner. Foaming of samples was abated with Dow Corning Antifoam A. For these analyses, a Beckman atomic absorption spectrophotometer system equipped with a 10-inch potentiometric recorder was used. Recovery rates were determined by adding mercury as HgCl to the various tissues. Data are corrected for recovery rates. The minimum detectable amount of mercury was 5 ppb. Accuracy has been given as 0.1 ppb (11) and 0.01 ng (8). Results and Discussion No heart tissue sample contained more than 5 ppb total mercury. The median mercury concentration in blood of birds killed on the day of capture was 7 ppb. Brain tissues of these first pigeons had a median residue level of 22 ppb; those caged for 1 week had only 6 ppb mercury in their brains. Subsequent samples of these tissues revealed no more than 5 ppb mercury (Table 1). Tejning determined that domestic fowls fed methyl- mercury-dicyandiamidc-treated grain excreted about 11 percent of the mercury (12). After cessation of experi- mental feeding, he observed that this loss gradually decreased and finally stopped altogether. Our data indi- cate that mercury was removed with feces in quantities of about 11 to 13 ppb for the birds. In captivity, how- ever, residues in excreta decreased to levels less than 5 ppb after 5 weeks. Feathers used in the study included mature and imma- ture contour and down feathers from the tail region. Pigeons that had been in captivity for 1 week showed somewhat higher concentrations of mercury in the Vol. 8, No. 2, September 1974 plumage than did those of subsequent analyses (Table 1). Mercury concentrations in feathers appear to show no consistent decline after the first week. Some variation in residue concentrations may be expected in natural populations of pigeons. Although analyses were not extended through a complete molting cycle, large con- centrations of total mercury in plumage seem to indicate that feathers are a pathway through which the metal is concentrated and eventually lost through molting. This corroborates Tejning's work on pheasants and domestic fowls which showed that most of the mercury in blood and organs is not excreted, but is transported to the plumage (12). Claws also contained sizable amounts of mercury (14-85 ppb). but no pathways were estab- lished with the limited data available although the residue level patterns in similar claws and feathers may suggest some correlation. Crops of pigeons killed on the day of capture contained corn, soybeans. Johnson grass seeds, and miscellaneous weed seeds. Other pigeons from the test birds' popula- tion were observed feeding along the right-of-way of a main-line railroad and in the yards of several grain stor- age facilities. Their drinking water was. for the most part, obtained from Town Creek, a generally sluggish stream receiving industrial waste and runoff that flows within 200 yards of the hotel. Mercury was undetectable in water samples from the creek. Pigeons drinking here have also been observed eating materials from bottom sediments near the shore of the creek. Crop contents of birds from the initial kill showed from less than 5 ppb to 7 ppb mercury. Authors suggest that possible con- tamination sources are treated grains, contaminated weed seeds, and naturally occurring mercurials. No apparently hazardous sources of mercury contamination were found to be available to these pigeons. A cknowledgments We thank Peter Lane. Jackson State University, for iden- tifying the plant materials in the crops of the pigeons, and Johnny Jackson, Jackson, Miss., for making the pigeons available for study. LITERATURE CITED (/) Lucas, H. F., Jr., D. N. EdincHon, nnd P. J. Colby. 1970. Concentrations of trace elements in Great Lakes fishes. J. Fish. Res. Bd. Can. 27(4) :677-684. (2) Fimreite, N. 1970. Mercury uses in Canada and their possible hazards as sources of mercury contamination. Environ. Pollut. H2):119-131. {.?) Kurland, L. T., S. M. Faro, and N. Sjcdeler. I960. Minamata disease. World Neurol. 1 (5) :36n-395. (4) Summer, A. K., ]. G. Saha, and Y. W. Lee. 1972. Mercury residues in fish from Saskatchewan waters with and without known sources of pollution — 1970. Pestle. Monit. J. 60:122-125. (5) Harriss, R. C. 1971. Ecological implications of mer- cury pollution in aquatic systems. Biol. Conser. 314): 279-283. (6) Fimreite, N., R. W. Fyfe, and J. A. Keiih. 1970. 103 Mercury contamination of Canadian prairie seed eaters and their avian predators. Can. Field Nat. 84(3):269- (10) 276. (7) Johnels, A. G., and T. Westermark. 1969. Mercury contamination of the environment in Sweden, pp. 221- 241, in M. W. Miller and G. Berg (Eds.) Chemical (11) Fallout. Charles Thomas Publishers, Springfield, 111. (8) Adley, F. £., and D. W. Brown. 1972. Mercury con- centrations in game birds, State of Washington — 1970 and 1971. Pestic. Monit J. 6(2):91-93. (12) (9) Benson, W. B., D. W. Brock. F. Shields III. E. R. Norherg. and J. Cline. 1971. An analysis of mercury residues in Idaho pheasants. J. Idaho Acad. Sci. Spec. Res. Issue. 2:17-26. Knight, L. A.. Jr., and J. Herring. 1972. Total mercury in largemouth bass (Micropteriis salmoides) in Ross Barnett Reservoir, Mississippi — 1970 and 1971. Pestic. Monit. J. 6(2): 103-106. Hatch, R., and W. L. Ott. 1968. Determination of submicrogram quantities of mercury by atomic absorp- tion spectrophotometry. Anal. Chem. 4O(14):2085- 2087. Tejning, S. 1967. Mercury in pheasants iPhasianus colcliicns L. ) deriving from seed grain dressed with meth\i and ethvl mercury compounds. Oikos. 18(1): 334-344. TABLE 1. Residues of total mercury (pph wet weight) in tissues and feces of pigeons from Jackson, Miss. — 1972 0 1 2 Weeks in Captivity 5 6 7 3 4 9 Brain Range 9-2.10 5-10 — — — — — — — Median 22 6 ND ND ND ND ND ND ND No. birds 9 3 3 3 3 3 3 3 3 Feattiers Range 8-.10 22-70 16-20 20-51 22-40 25-26 <5-26 21-33 14-21 Median 19 49 17 22 27 26 13 30 18 No. birds 10 3 3 3 3 3 3 3 3 Skin Range 5-44 20-49 6-16 <5-5 <5-7 5-6 — <5-13 <5-6 Median 29 .15 10 5 6 5 ND 11 ND No. birds 7 3 3 3 3 3 3 3 2 Scales Range — 14-15 6-10 <5-ll 8-18 5-10 — 11-19 <5-58 Median ND 15 8 5 10 5 ND 11 8 No. birds 9 3 3 3 3 3 3 3 3 Liver Range — <5-9 — — <5-5 5-10 — — — Median ND 6 ND ND ND 5 ND ND ND No. birds 9 3 3 3 3 3 3 3 3 Breast Range <5-51 — — <5-5 6-9 — — — — Median 20 ND ND 5 9 ND ND ND ND No. birds 7 3 3 3 3 3 3 3 3 Bones Range <5-6 — — — 8-16 1(1-17 — — — Median ND ND ND ND 8 15 ND ND ND No. birds 9 3 3 3 3 2 3 3 3 Blood Range 5-12 — — — — — — — — Medi.in 7 ND ND ND ND ND ND ND ND No. birds 10 3 2 2 2 2 1 2 3 Heart Range Median ND ND ND ND ND ND ND ND ND No. birds .s 1 3 3 3 3 1 3 3 Claws 1 29 59 34 55 14 51 23 47 85 No . birds 20 3 3 3 3 3 3 3 3 Oil Glands ' 10 29 NS 39 84 — 50 8 13 No. birds 20 3 NS 3 3 3 3 3 3 Feces ' 11 13 11 9 7 NS — — — No. birds 20 3 3 3 3 NS 3 3 3 NOTE: — = concentrations of mercury less than 5 ppb. ND :n: not determined. NS = no sample. 1 Tissues or feces combined for analysis. 104 Pesticides Monitoring Journal Distribution of PCB and p,p'-DDE Residues in Atlantic Herring (Clupea harengus harengus) and Yellow Perch (Perca flavescens) in Eastern Canada — 1972^ V. Zitko, p. M. K. Choi, D. J. Wildish, C. F. Monaghan, and N. A. Lister ABSTRACT In two localities of Eastern Canada in 1972, Atlantic herring (Clupea harengus harengus) and yellow perch (Perca flaves- cens) were analyzed for residues of poly chlorinated hiphenyls (PCB's), p.p-DDE, p,p'-DDD. and p.p'-DDT, and for lipid and body weight. From eacli locality. 24 herring and 25 perch were analyzed: all were males between 2 and 4 years old, and all were analyzed individually. Data were statistically evaluated. In most cases PCB, p,p'~DDE, and lipid were belter approximated by lognormal distribution; weight was better approximated by normal distribution. All determined parameters were highly variable. One objective of the study was to weigh the possibility of using the two species under observation to detect temporal trends in levels of environmental contamination by PCB's and p,p'-DDE. The study concludes, however, that it is probably impossible to detect a significant change of PCB levels in fish of equal age in 2 consecutive years by using samples of this size. Introduction This paper describes levels and distribution of poly- chlorinated biphenyls (PCB's), p,p'-DDE. wet body weight, and lipid in Atlantic herring (Clupea harengus harengus) and yellow perch (Perca flavescens). An ob- jective of this study was to evaluate the possibility of using these species to detect temporal trends in levels of environmental contamination by PCB's and p.p'-DDE. Environment Canada, Biological Station, St. Andrews, New Bruns- wick, Canada. Species originated in waters of Eastern Canada: Gulf of St. Lawrence. Bay of Fundy, and New Brunswick Rivers. Forest-based industries, mining, and smelting opera- tions are the typical industrial activities in the Gulf of St. Lawrence area. In the past. DDT was used for forest spraying in New Brunswick. Quebec, and Newfound- land; other pesticides are still used for agricultural pur- poses on Prince Edward Island. The St. Lawrence River runoff may be a source of chlorinated hydrocarbons. Little industrial or agricultural activity takes place in southern Nova Scotia. Surface circulation in the Gulf of Maine — Bay of Fundy area (/) may bring chlori- nated hydrocarbons from the south into the area. Occurrence and movement patterns of four major stocks of Atlantic herring (2) are illustrated in Figure 1. Levels of PCB's and chlorinated hydrocarbon pesticides in two weight groups of the Banquereau stock and in one weight group of the Gulf of Maine stock have been reported (3). Pooled samples were analyzed in the earlier study; there was no information available on statistical distribution of the residues with which to compare results of the current study. The concentration of PCB's and chlorinated hydrocarbon pesticides in her- ring oils obtained by commercial processes from the Gulf of St. Lawrence herring stock between 1967 and 1970 was determined by Addison et al. (4). Yellow perch from New Brunswick. Canada, has not been analyzed previously. Vol. 8, No. 2, September 1974 105 Materials and Methods SAMPLING Samples of herring were obtained from commercial fish- eries. Sex, age. and wet body weight were determined: male fish were frozen ( — 14°C) until analysis. Herring sample B (Gulf of St. Lawrence stock) was taken May 25. 1972; herring sample T (Nova Scotia stock) was TABLE L Residues of PCB's in muscle of Atlantic herring and yellow perch — Eastern Canada, 1972 Residues, ^g/g Wet Weight Standard Chi square Sample i Mhan DEVIATION PROBABILITY. ^ N L N L N L Herring B 0.31 0.25 0.18 0.27 2 50 Herring T 0.64 0.44 0.49 0.43 5 t Perch 2 0.089 0.078 0.031 0.18 50 70 Perch 3 0.13 0.12 0.046 0.22 20 80 NOTE: N = data as such; L = logarithmically transformed data. Mean is antilog form; standard deviation is in log form. ' Herring samples represented 24 individual fish; nerch represented 25. TABLE 2. Residues of p,p'-DDE in muscle of Atlantic herring and yellow perch — Eastern Canada, 1972 Residues, yg/g Wet Weiciti Sample '•= Mean Standard deviation Chi square probability, % N L N L N L Herring B Herring T Perch 3 0.070 Oil 0.11 0 059 0.0H5 0 089 1) 048 0,071 0.040 0 2" 0.43 0.22 Id 1 21) 70 70 NOTE: N = data as such; L — logarithmically transformed data. Mean is antilog form; standard deviation is in log form. ' Herring samples represented 24 individual fish; perch represented 25 2 DDE not detectable in perch sample 1. TABLE 3. Residues of p,p'-DDD and p,p'-DDT in muscle of Atlantic herring — Eastern Canada, 1972 Residues, ug/g Wet Weight ' Mean Standard deviation Sample = P.p'-DDD p.p'-DDT P.p'-DDD p.p'-DDT Herring B Herring T 0.042 0.049 0.021 0.041 0.31 0.27 0.37 0.44 ' Logarithmically transformed data. Mean is in antilog; standard de- viation is in log form. '^ Herring samples represented 24 mdividual fish; perch represented 25. TABLE 4. Hexane-extractable lipid in muscle of Atlantic herring and yellow perch — Eastern Canada, 1972 Lipid, % Wet Weight Standard Chi square Sample ' Mean deviation PROBABUITY. '7c N L N L N L Herring B 2.70 1.9S 1.93 0.42 -> 30 Herring T 12.7 11.7 6.26 0.38 50 5 Perch 2 0.32 0.29 0.17 0.21 20 30 Perch 3 0.54 0.51 0.20 0.17 1 20 taken August 18, 1972 (Tables 1-7). The former fish are spring spawners and the latter are fall spawners; both samples were taken just before spawning. Herring in these samples were 4 years old. TABLE 5. Body weight of Atlantic herring and yellow perch — Eastern Canada, 1972 Sample > Arithmetic mean - Standard deviation Chi square probability, % Herring B Herring T -i Perch 2 188 194 9.3 15 0.066 2.34 50 30 80 NOTE: Weights not available for perch sample 3. ' Herring samples represented 24 individual fish; perch represented 25. - Grams, wet weight. ^ Logarithmically transformed data. Mean is in antilog form, standard deviation is in log form. TABLE 6. Decrease of PCB and p,p'-DDE residues in muscle of Atlantic herring and yellow perch — Eastern Canada Sample i New mean in % OF PRESENT MEAN PCB P.P' DDE Distribution Distribution N 1. N L Herring B 72 76 81 73 Herring T 62 64 73 62 Perch 2 83 82 ND ND Perch 3 85 79 82 79 NOTE; N = data as such; L = logarithmically transformed data. Mean is in antilog form: standard deviation is in log form. ' Herring samples represented 24 individual fish; perch represented 25. NOTE: N = normal; L = lognormal; ND = no residues detected. Data are statistically significant at 5% probability. ' Herring samples represented 24 individual fish; perch represented 25. Yellow f)erch were obtained by seining from the shore with a 3-ft-by-15-ft chain-weighted net with floats and a mesh size of '2 inch; Figure 1 indicates sampling stations. Se.x, age, and wet body weight were deter- mined and, like the herring, male perch were frozen (— 14°C) until analysis. Perch sample 2 was obtained May 2 and 5, 1972; perch sample 3 was obtained May 25, 1972. Analysis was performed on two- and three- year-old fish from stations 2 and 3, respectively. Station 2 (Grand Falls, St. Croix River) is in a rather sparsely populated area with little agricultural or industrial ac- tivity. A pulp mill and two towns with a total population of about 12,000 are located downstream. Station 3 (Grand Point, Grand Lake) is on the St. John River, where pulp mills, potato and food processing plants, and a system of dams are located. The station is about 30 miles downstream from the city of Fredericton (pop- ulation 25,000), and 40 miles upstream from the city of Saint John (population 100,000). ANALYSIS Sample analysis similar to that described in a 1972 study (5) utilized the white lateral muscle between the dorsal fin and the lateral line. The sample (5.5-6.0 g for her- ring, 1.5-2.0 g for perch) was ground with anhydrous sodium sulfate (Fisher Scientific S-421, 30 g) in a mor- 106 Pesticides Monitoring Journal TABLE 7. Residue levels of PCB's and p,p'-DDE (iig/g wet weight), hexane-extractahle lipid (% wet weight), and body weight (g, wet weight), in Atlantic herring and yellow perch — Eastern Canada, 1972 PCB p,p'-DDE 1 Lipid Weight PCB P.P-DDE Lipid Weight Herring B i Herring T i 0.51 0.18 1.74 202.8 1.02 0.16 13.7 186.7 0.21 0.04 2.15 191.3 0.60* 0.11* 10.1* 210.9 0.18 0.04 1.53 183.2 0.50» 0.09* 9.38* 0.44 0.10 0.04* 4.75 0.34* 180.2 171.7 0.15* 0.13* Perch 2 = 0.04* 0.32* 0.22 0.06 2.65 182.1 0.11 TR 0.13 11.51 0.17 0.04 0.81 191 0 0.12 TR 0.59 6.92 0.11 0.03 0.42 177.1 0.08 TR 0.14 6.98 0.28 0.06 5.24 196.8 0.07 0.01 0.16 10.10 0.16* 0.05* 0.96* 177.1 0.09 TR 0.41 7.32 0.15* 0.04* 0.95* 0.16 TR 0.19 6.65 0.24 0.06 3.27 178.9 0.08 0.01 0.25 7.38 0.51 0.06 1.46 198.4 0.07 TR 0.26 9.34 0.55 0.14 4.76 178.7 0.05 TR 0.28 10.10 0.70 0.19 3.52 192.5 0.22 TR 0.44 7.91 O.IO* 0.02* 0.88* 189.1 0.12 TR 0.31 8.73 0.12* 0.03 • 1.02* 0.05 TR 0.28 10.40 0.40 0.09 1.64 192.5 0.04 TR 0.29 13.41 0.26 0.06 4.30 200.3 0.09 TR 0.50 5.65 0.41 0.07 12.94 146.4 0.07 TR 0.34 9.32 0.10 0.03 0.43 164.5 0.06 TR 0.76 9.49 0.12' 0.03* 1.32* 196.7 0.05 O.OI 0.32 13.16 0.12* 0.03 • 1.64* 0.06 TR 0.30 9.74 0.56 0.16 5.68 210.8 0.08 TR 0.29 10.91 0.42 0.11 4.88 215.1 0.07 TR 0.19 8.27 0.46 0.15 4.91 186.2 0.05 TR 0.23 13.04 0.09* 0.03 • 0.48* 208.3 0.09 0.01 0.14 8.95 0.12* 0,02« 0.64* 0.10 0.05 0.01 TR 0.34 0.29 8.04 12.15 Herring T i 0.12 TR 0.74 5.72 0.02 0.03 6.28 131.5 0.08 0.19 Perch 3 » 6.71 145.0 0.20 0.04 6.95 218.7 0.09 0.13 1.42 O.IO 0.02 5.00 177.2 0.14 0.10 0.87 0.65* 0.16* 16.7* 212.3 0.04 0.05 0.39 0.61 • 0.14* 16.0' 0.05 0.05 0.67 0.12 0.03 7.87 201.8 0.15 0.10 0.47 0.13 0.03 9.07 213.2 0.06 0.08 0.69 0.15 0.04 3.39 187.2 0.11 0.14 0.33 2.28* 0.32* 26.9« 233.4 0.18 0.17 0.35 2.17* 0.32« 24.3* 0.27 0.19 0.70 0.03 0.01 0.69 176.1 0.11 0.07 0.57 0.56 0.11 11.5 202.1 0.14 0.04 0.56 1.58 0.40 22.4 224.5 0.29 0.13 0.91 1.29 0.29 19.6 229.4 0.26 0.24 0.47 0.48* 0.09* 16.1* 163.2 0.14 0.10 0.40 0.53 • 0.09* 15.8* 0.10 0.06 0.35 0.61 0.11 20.1 197.7 0.12 0.09 0.88 0.62 0.11 17.1 175.0 0.22 0.15 0.44 0.44 0.09 15.4 163.0 0.07 0.05 0.37 1.07 0.20 15.8 202.0 0.09 0.13 0.10 0.14 0.34 0.44 0.57* 0.07* I6.5* 181.4 0.10 0.09 0.31 0.55* 0.07* 16.1» 0.07 0.08 0.37 1.62 0.29 12.1 216.8 0.13 0.03 0.41 0.88 0.15 18.2 244.6 0.11 0.07 0.48 1.04 0.18 12.9 210.1 0.19 0.05 0.50 NOTE: • =: duplicate analyses of a single specimen. TR : 1 Herring samples represented 24 individual lish. " Perch samples represented 25 individual fish. ^ Weights not available for perch sample 3. : trace: <0.01. tar to yield a free-Plowing powder and extracted with pesticide-grade hexane (Fisher Scientific H-300) in a So.xhlet extractor for I hour. The volume of the extract was adjusted to 100 ml in a volumetric flask with pesticide-grade hexane. An aliquot of the extract (10- 20 ml) was dried in a rotatory evaporator in vacuum at room temperature to determine hexane-extractable lipids. An aliquot, not exceeding 100 mg of lipids, was applied in 1 .5 ml pesticide-grade hexane to a 45-by- 0.7-cm glass column with a glass wool plug, containing 2 g alumina. Fisher Scientific A-540, activated at 800°C for 4 hours and deactivated by the addition of 5 per- cent distilled water. The solvent was washed into the column with another 1.5 ml pesticide-grade hex- ane and the column was percolated with the same sol- vent to collect 20 ml effluent. After the effluent had been concentrated almost to dry- ness on a rotatory evaporator, it was applied in 1.5 ml 0.5 percent v/v pesticide-grade benzene (Fisher Scientific B-426) in pesticide-grade hexane to a column Vol. 8, No. 2, September 1974 107 FIGURE 1. Atlantic herring stock structure and yellow perch sampling stations — Eastern Canada. 1972 filled with silica. The column was the same as that de- scribed above, and contained 2 g Mailinckrodt Silica- 100-200 mesh. The solvent was washed with pesticide- grade hexane, dried in a rotatory evaporator at 36°C in vacuum, activated overnight at 130°C, and deactivated by the addition of 3 percent distilled water. The apphed aliquot was washed into the column with another 1.5 ml of the solvent. The column was perco- lated with the same solvent to collect 1 5 ml effluent. This fraction contained PCB's and p,/?'-DDE. The column was percolated further with 10 percent diethyl ether (Fisher Scientific E-134) in hexane and 10 ml effluent was collected. This fraction contained p.p'-DDD and /j.^'-DDT. Fractions were evaporated just to dryness on a rotatory evaporator in vacuum at room temperature. The residue was dissolved in pesti- cide-grade hexane (0.2-0.4 ml) and analyzed by gas chromatography. The gas chromatograph, a Packard A7901, was operated at 200°C. It had a glass column (6 ft by 4 mm), containing 4 percent SE-30 on acid- washed Chromosorb W, 60-80 mesh. Carrier gas was nitrogen at a flow rate of 60 ml/min. Injector and de- tector (^H, 150 mc) were kept at 210°C. A solution of Aroclor 1254 (2.232 |.ig/ml) and a solution containing p.p'-DDE, p,p'-DDD, and p.p'-DDT (0.182, 0.235, and 0.247 Jig/ ml, respectively) were used daily to calibrate the detector. Heights of five of the six major peaks of Aroclor 1254 were used for quantitation. Peak heights were also used to quantitate the other compounds. Mini- mum detectable levels of PCB's, p.p'-DDE, ;),p'-DDD. and /),/)'-DDT were 0.01, 0.003, 0.002, and 0.002 iig/g wet weight, respectively. All glassware was washed with a laboratory detergent in tap water and rinsed with distilled water, acetone, and pesticide-grade hexane. Sodium sulfate was washed with pesticide-grade hexane and dried in a rotatory evaporator at 36°C in vacuum. Extraction thimbles and glass wool were pre-extractcd with pesticide-grade hex- ane in a Soxhlet extractor. In preparing the 0.5 percent v/v benzene in pesticide-grade hexane, an allowance was made for benzene already present in hexane. The concentration of benzene in each batch of pesticide- grade hexane was determined from the absorbence at 253 nm against spectrogradc hexane (Fisher Scientific H-334). In each sample, 24 herring and 25 perch were analyzed individually. Five fish from each sample were analyzed in duplicate, including duplicate extraction. Results of duplicate analyses usually agreed within 10 percent and arithmetic means of duplicate analyses were used in further evaluation of data. DATA EVALUATION Arithmetic means and standard deviations (Tables 1-6) were calculated from the data (Table 7) and from their decadic logarithms. To determine whether distribution of data can be approximated by a normal or a lognormal distribution, data were divided into classes; enough classes were chosen to assure that the original mean and standard deviation would not alter appreciably. Frequencies expected for normal and lognormal distri- bution were calculated from the means and standard deviations and compared with observed frequencies, testing the goodness of fit by the chi square test (6). Higher chi square probabilities (Tables 1,2,4,5) indi- cate better fit of the theoretical to the observed distri- bution. Results and Discussion PCB'S In all samples PCB's resembled and were quantitated as Aroclor 1254. Data are summarized in Table 1. As can be seen from the chi square probability, in three out of four cases lognormal distribution would be preferred to normal. PCB distribution in the herring sample T could not be adequately characterized by either distri- bution. The PCB level reported previously (3) for the Banquereau stock of similar weight group (arithmetic mean 0.54) falls between the levels observed in the Nova Scotia and Gulf of St. Lawrence stocks. This may indicate that the level of PCB's in herring of equal size decreases in the northerly direction. CHLORINATED HYDROCARBON PESTICIDES levels of /).//-DDF, arc presented in Table 2. DDE was not detectable in perch sample 2. In all cases, the dis- tribution is better described as lognormal. Levels of p.p'-DDD and p.p'-DDT in herring are summarized in Table 3. Because of the low levels found, no detailed statistical evaluation of the data was made and it was assumed that, as in the case of p.p'-DDE. the distribu- tion is lognormal. No measurable residues of p.p'-DT>D and p.p'-DDT were found in perch sample 2. Perch sample 3 contained very low levels of these compounds 108 Pesticides Monitoring Journal (0.005-0.02 i^ig/g) in six fish analyzed individually. Statistical evaluation was not carried out. LIPID AND BODY WEIGHT In three out of four cases the distribution of lipid was best described as lognormal (Table 4). Herring sample T may be described by a normal distribution. This was the only sample taken at the end of the feeding season and it is possible that there is a relation between the feeding activity and the distribution of lipid. The weight of the fish was normally distributed (Table 5), with the exception of herring sample T. where the data may be described by a lognormal distribution. Weights were not available for perch sample 3. TRENDS OF PCB AND p,p'-DDE LEVELS Knowing the trends of the environmental levels of per- sistent pollutants enables one to assess the effectiveness of regulatory actions such as the Canadian ban on DDT and restriction of PCB's. As stated earlier, one objective of this study was to weigh the possibility of using Atlantic herring and yellow perch to detect such trends. Authors' findings now render that possibility obscure. Analytical data indicate that even after elimination of possible sex and age (or size) variations, the variability between individual fish still remains very high and a large number of fish have to be analyzed to detect sta- tistically significant differences. Table 6 calculates dif- ferences for the 5 percent probability level, using the observed standard deviations and assuming that 25 indi- vidual fish are analyzed in each sample. It is very likely that no significant difference in PCB levels can be detected in 2 consecutive years when using samples of this size. Concentration of PCB's in fish is determined by the rate of PCB uptake and excretion and by the growth of fish. The rate of excretion is very slow. In related studies fish did not metabolize chloro- biphenyl isomers (7) and excreted very little, if any, Aroclor 1254 ingested in a laboratory experiment (8). If, in a hypothetical case, the uptake of PCB's were completely eliminated, concentration of PCB's in fish would depend only on the rate of growth. The relative weight increase of the investigated stocks of herring between the age of 3 and 4 years is approximately 46 percent and the level of PCB's in 3-year-old fish is about 86 percent of that in 4-year-old fish (9). The con- centration of PCB's in 4-year-old herring in 1974 would therefore be 59 percent of the level in herring of the same age in 1973. This hypothetical level is quite close to the minimum detectable difference in Table 6. The uptake of PCB's cannot be completely eliminated due to the presence of PCB's in the environment; the dif- ference will be smaller than estimated above. It may be more pertinent and less costly to analyze a large num- ber of individual fish only every 4-5 years to determine trends of PCB levels. Insufficient data on the excretion rate of p.p'-DDE by fish do not allow prediction of trends of p.p'-DDE levels. It is possible that p,p'-DDE residues disappear from fish faster than PCB residues. The average level of pp' -DDE in the Banquereau herring in 1971 was 0.24 |(g/g (3) and the levels of p.p'-DDE in herring oils in 1968-70 were comparable to those of PCB's (4). Levels of p.p'-DDE reported in this paper are much lower. A cknowledgments We thank Madelyn M. Irwin for typing the manuscript and P. W. G. McMullon and F. B. Cunningham for drawing Figure 1 . LITERATURE CITED (/) Bumpus. D. F., and A/. Lauzier. 1965. Surface cir- culation on the continental shelf off Eastern North America between Newfoundland and Florida. Serial Atlas of the Marine Environment Folio 7. American Geographical Society. (2) lies, T. D., and S. N. Tihbo. 1970. Recent events in Canadian Atlantic herring fisheries. ICNAF Redbook, Ft. Ill: L34-147. (.?) Ziiko. V. 1971. Polychlorinated biphenyls and organo- chlorine pesticides in some freshwater and marine fishes. Bull. Environ. Contam. Toxicol. 6:464-470. (4) Addison. R. F.. M. E. Zinck. and R. G. Ackman. 1972. Residues of organochlorine pesticides and poly- chlorinated biphenyls in some commercially produced Canadian marine oils. J. Fish. Res. Board Can. 29(4): 349-355. (5) Ziiko, V. 1972. Problems in the determination of polychlorinated biphenyls. Intern. J. Environ. Anal. Chem. 1:221-231. (6) Sokal. R. R.. and F. J. Rohlf. 1969. Biometry. The principles and practice of statistics in biological re- search, pp. 550-572. W. H. Freeman and Company, San Francisco, Calif. (7) Hiilzini^er. O.. D. M. Nash, S. Safe. A. S. W. DeFreilas, R. J. Norsirom. D. J. Wildi.ih, and V. Ziiko. 1972. Polychlorinated biphenyls: Metabolic behavior of pure isomers in pigeons, rats, and brook trout. Science 178:312-314. (S) Zitko, v., and O. Hutzinger. 1973. Sources, levels, and toxicological significance of PCB's in hatchery-reared Atlantic salmon, in PCB's — still prevalent — still persis- tent. C. G. Gustafson, ed., Marcel Dekker, Inc., New York, N.Y., in press. (9) Monaghan, C. F., and V. Zitko. 1973. Unpublished data. Vol. 8, No. 2, September 1974 109 RESIDUES IN FOOD AND FEED Pesticide Residues in Total Diet Samples (VII) D. D. Manske ' and P. E. Corneliussen '^ ABSTRACT Pesticide residue levels delected in ready-to-eat foods re- mained at relatively low levels during the seventh year of the Total Diet Study in its present form. Samples were collected from 30 markets in 27 different cities. Popula- tions of cities ranged from less than 50.000 to 1.000.000 or more. Averages and ranges of pesticides commonly found are reported for the period June 1970-April 1971 by region and food class. Pesticides found infrequently are also reported for tliis period hy region and food class. Results of recovery studies with various classes of pesticides are also presented. After October 1970, analy.u-s of bromides, amitrole. and dithiocarbamates were discontinued: mercury and orthophenylphenol were added. Residue levels in three major fatty food groups are now reported on a whole- product basis, rather than on a fat basis. Data for June and August were adjusted accordingly. IiUrodiiclion The Total Diet Program (/) has been used since 1964 by the Food and Drug Administration (FDA), U.S. Department of Health. Education, and Welfare, as a way to monitor pesticide residues in foods. This pro- gram measures the amount of pesticide chemicals in food samples collected in retail outlets and prepared for consumption. Although the program was designed to measure pesticide residues in foods, some chemicals determined and reported here may not have been used as pesticides; this includes such materials as polychlori- nated biphenyls (PCB's), cadmium, and mercury. In- creased awareness of the potential hazards presented by these chemicals necessitates their inclusion in this pro- gram. ^ Kansas City Field Office Lal^oratory, Food and Drug Administration. U.S. Department of Health, Education, and Welfare, Kansas City, Mo. 64106. 2 Division of Chemical Technology, Bureau of Foods, Food and Drug Administration, U.S. Department of Health, Education, and Welfare, Washington, D.C. 20204. Amounts and types of residues found from June 1964 through April 1970 have been described in earlier re- ports f--7). The present report covers the period June 1970 through April 1971. Tabular data included are comparable to those of previous years. A Italy tical Methods No significant changes were made in sampling and com- positing procedures described in the initial issue of Pesticides Monitoring Journal (I). Samples were col- lected in 30 different grocery markets in 27 different cities representing five regions of the United States: Baltimore, Boston, Kansas City, Los Angeles, and Min- neapolis, Population of the cities ranged from less than 50.000 to 1,000,000 or more; the average sampling site was in the 250,000 to 500,000 range. Beginning with the October 1970 sample, significant changes were made regarding analysis. At that time, all Total Diet analyses were centralized at the FDA Kansas City district. Analyses for bromides, amitrole, and dithiocarbamater, were discontinued, and PCB's, mer- cury, and orthophenylphenol (o-phenylphenol) were added. Analyses for residues of organochlorines, some parent organophosphorus pesticides, chlorophenoxy acid herbicides, carbaryl, arsenic, and cadmium were continued as in previous programs. Procedural changes were made in analyses of fatty food groups for chlorinated and organophosphorus residues. These food groups were Group I: Dairy Products; Group II: Meat, Fish, and Poultry; and Group X: Oils, Fats, and Shortening. Prior to October 1970. all organo- chlorine and organophosphorus residues found in these three groups were reported on a fat basis. After this date, residues were calculated on a whole-product basis. Results reported here from June and August samples have been converted to the whole-product basis using 110 Pesticides Monitoring Journal an average fat content of 10.5 percent for Group I, 20 percent for Group II, and 85.5 percent for Group X. Methods used for most analyses are described in the FDA Pesticide Analytical Manual, Vol. I and II f^). Other analytical methodology was used for certain resi- dues: colorimetry for arsenic (9)\ flameles^ atomic ab- sorption spectroscopy for mercury (JO), and atomic absorption or polarography for cadmium (II). Carbaryl and o-phenylphenol were extracted by the method of Porter et al. (12} and quantified by thin-layer chroma- tography (TLC) (13). Quantitative values reported for organochlorine and organophosphorus compounds were obtained by gas- liquid chromatography (GLC) using the electron cap- ture, the thermionic or the flame photometric detection system. Confirmation was made by TLC and/or GLC with halogen-specific microcoulometric detection. For additional confirmation of residues, identity techniques such as p-values, alternative GLC column packings, alkaline hydrolysis, and mass spectrometry were used. The methodology permits the quantitation of heptachlor epoxide, an organochlorine, at about 0.003 parts per million (ppm). and of parathion. an organophosphate, at about 0.01 ppm. The limit of quantitation for both classes varies with the individual compound being de- termined. In some cases, finite residue levels are re- ported below the approximate limits of quantitation. The quantitative reliability of these extremely low levels is not known because the authors have no recovery information about dietary composites below 0.003 ppm for organochlorine compounds or 0.01 ppm for organo- phosphorus compounds. As a general rule residue de- terminations below 0.005 ppm are subject to a high degree of variability. Those residues detected and quali- tatively confirmed at too low a level to be quantified are reported as trace (T). Approximate limits of quantitation for other analyses were chlorophenoxy acid herbicides and pentachloro- phenol (PCP): 0.02 ppm; carbaryl: 0.05 ppm; arsenic: 0.1 ppm; cadmium: 0.01 ppm: amitrole (June and August samples only): 0.05 ppm: bromides (June and August samples only): 0.5 ppm. Individual fruits and vegetables were examined for dithiocarbamate residues (June and August samples only). Beginning in October 1970, each composite was examined for PCB's with a limit of quantitation of about 0.05 ppm: for o-phenyl- phenol, with a limit of O.I ppm; and for mercury, with a limit of 0.02 ppm. Residts A total of 1,081 residues of 33 different chemicals were found in samples in the current reporting period. In the previous reporting period, 1,446 occurrences of 31 different chemicals were found. This apparently large decrease is deceptive; almost one-half the decrease oc- curred in reported bromide residues because bromide analyses were discontinued in October 1970. As in pre- vious years, a total of 360 composites were examined for all residues; there were 12 composites from each of the 30 markets. The program calls for carbaryl and o-phenylphenol analyses on only the nonfatty compos- ites (1,3-6.9-1]. 13), of which there were 270. Pro- gram changes also resulted in analyses of only 240 composites for mercury and 120 composites for bro- mides. The 33 different residues found are listed in decreasing order of frequency in Table 1 . The most common residues, maximum levels of those residues, and residues reported less frequently are dis- cussed below for each of the 12 food composites. Tables 2a and 2b report findings in more detail accord- ing to food class and region. None of the reported find- ings have been corrected for recoveries obtained in recovery experiments. Table 3 summarizes studies. DAIRY PRODUCTS Of the 30 composites examined, 28 contained residues. Organochlorine pesticide residues were the most com- mon, appearing in 27 composites. The most common organochlorines and their maximum concentrations were DDT. 0.005 ppm; DDE, 0.028 ppm; dieldrin, 0.007 ppm; heptachlor epoxide, 0.00! ppm; and BHC, 0.001 ppm. Also found were hexachlorobenzene (HCB), lindane. TDE, metho.xychlor, and PCP. Bro- mides were found in 7 of 10 composites examined at levels of 1.0 ppm to 5.5 ppm. Cadmium was found in 4 of 30 composites at 0.01 to 0.06 ppm. MEAT, FISH. AND POULTRY Residues of 9 organochlorine compounds were found in varying combinations in all 30 composites examined. Most common organochlorine residues and their maxi- mum concentrations were DDE: 0.048 ppm; DDT: 0.033 ppm; TDE: O.OIO ppm; and dieldrin: 0.015 ppm. Heptachlor epoxide, lindane. BHC, bromides, and HCB were also found, although less frequently. PCB's were found in 14 of the 30 composites; tlie highest PCB residue was 0.15 ppm. Arsenic was found in 9 compos- ites: 0.1-0.3 ppm; and cadmium was found in 17 com- posites: 0.01-0.04 ppm. GRAIN AND CEREAL PRODUCTS Organophosphorus residues were the most common found in this commodity class. Malathion was found in 28 of 30 composites (maximum level 0.170 ppm): diazinon was found in 12 of 30 composites (maximum level 0.015 ppm). Varying combinations of 8 organo- chlorine compounds were found in 20 of 30 composites. The most common organochlorines and their maximum levels found were DDE: O.OOI ppm; DDT: 0.009 ppm; and dieldrin: 0.012 ppm. PCB's were found in 4 composites; 0.36 ppm was the highest level reported. Vol.. 8, No. 2, September 1974 111 Cadmium was found in 27 of 30 composites at O.OI- 0.07 ppm. Other residues found were TDE. ronnel, o-phenylphenol, and metlioxychlor. POTATOES Residues of 8 organochlorine compounds were de- tected in 15 of the 30 composites. The most common of TABLE 1. Insecticide residues found in food composites, June 1970-April 1971'' No. OF Positive No. OF Composites with Ranges Composites Residues Reported of PPM Chemical Found WITH Residijes As Trace = Reportinos CADMIUM 213 0 0,01-0.20 DDE l,l-dichloro-2,2-bis (p-ch!orophenyl) ethylene (isomers other than p.p' also included in reportings) 133 55 T-0.028 DDT l,l,l-trichloro-2,2-bis (p-chlorophenyl) ethane (isomers other than p,p' also included in reportings) 118 51 T-0.037 DIELDRIN Not less than 85% of l,2..1,4.10.1()-hexachloro-6,7-epoxy-l,4,4a.5,6,7,8.8a- octahydro-l,4-fn(^o-fxo-?.8-dinielhanonaphthalene 110 23 T-0.015 TDE l,l-dichloro-2.2-bis (p-chlorophenyl) ethane (isomers other than p.p' also included in reportings) 90 42 T-0.087 BROMIDES ' 81 0 0.5-51 MALATHION diethylmercaptosuccinate, 5-ester with O.Odimethyl phosphorodithioate 49 2 T-0,170 BHC 1.2,3.4,5.6-hexachlorocyclohexane, mixed isomers except gamma 42 19 T-0.009 HEPTACHLOR EPOXIDE 1.4,5.6.7,8,8-heptacliIoro-2..1-epoxy-3a.4,7.7a-tetrahydro-4,7-melhanoindaii 36 23 T-0.01 1 LINDANE 1,2,3,4,5.6-hexachlorocyclohexane, 99% or more gamma isomer 28 8 T-0.023 ENDOSULFAN 6.7,8.9,lt1.l0-hexachloro-l.5.5a,6.9,9a-hexahydro-6,9-methano-2.4,3- benzodioxalhiepin 3-oxide (leportings include isomers I. II, and the Sulfate) 24 R T-0,063 CARBARYL ' I-naphthyl methyl carbamate 20 15 T-0.5 PCB'S (polychlorinated biphenyls) Calculated as Aroclor'^ with varied chlorine content — 54% and 60% reported Ibis period 18 13 T-0.36 DIAZINON 0.0-diethyl 0-(2-isopropyl-6-methyl-4-pyrimidinyl ) phosphorothioate 15 10 T-0.015 ETHION O.O.O'.O'-tetraethyl 5,5'-methylene bisphosphorodithioate 15 3 T-0.099 ARSENIC (As=0,i) 13 0 0.1-0.3 DICOFOL (KELTHANE*! 4,4'-dichloro-a-(trichIorometlivl ) benzhvdrol 13 2 T-0.066 PARATHION 0.0-diethyl 0-p-nitrophenyI phosphorothioate 11 4 T-0.01 2 ORTHOPHEN-^I PHENOL ' 2-bvdroxvdiphenyl 10 7 T-0.25 MERCURY ■ 10 3 T-0.05 HCB hexachlorobenzene 6 3 T-0.(I01 PERTH ANE l.l-dichIoro-2.2-bis (p-elhyl phenyl) ethane 5 2 T-0.022 METHOXYCHLOR l,l,l-trichloro-2,2-bis (p-melhoxyphcnyl) ethane 4 2 T-0.008 METHYL PARATHION 0,0-dimethyl (?-p-nitrophenyl phosphorothioate 2.4-D 2, 4-dichlorophenoxy acetic acid 3 2 T-0.01 1 0 0.01-0.02 PENTACHLOROPHENOL 1 T-0.01 1 BOTRAN9 2,6-dicliloro-4-nitroaniline 0 0.003-0.006 ENDRIN 1,2, 3.4, 10.I0-hexachloro-6.7-cpoxy-l, 4,4a, 5.6,7. 8,8a-octahydro-l,4- en(/o-eft(io-5,8-dimethanonaphlliaIene 1 T0.007 TOXAPHENE chlorinated camphene containing 67-69% chlorine 1 T DCPA (DACTHAL*) 2.3,5.6-tetrachlorotercphthalic acid dimethyl ester 1 T CIPC isopropyl n-(3-chlorophcnyl ) carbamate 0 0 14 PHOSALONE .?l(6-chloro-2-oxo-3-benzoxa/olinvl ) | metliyl rt,»-diethvl phosphoroduhioatc 1) 1)278 RONNEL 0,0-dimethyI 0-2,4,5-triclilorophenyl phosphorothioate 0 0.007 mercury — 240 composites; bromides — 120 composites; carbaryl and Total of 360 composites examined for each compound except as follo' orthophenyl phenol — 270 composites. Pesticide chemicals capable of belny detected by the specific analytical methodology may be confirmed qualitali when they are presciH at concenn aiion^ below the limir of quantitation 1 imit oi quantitation varies with residue vcly and but are not quantifiable food class. 112 Pesticides Monitoring Journal these were dieldrin, DDE, and DDT with maximum values of 0.006 ppm. 0.018 ppm, and 0.020 ppm, re- spectively. Also detected were Botran (dichloran), TDE, BHC, endrin, carbaryi. endosulfan, CIPC (chlor- propham), bromides, and arsenic. Cadmium was found in 28 of 30 composites at 0.02-0.09 ppm. LEAFY VEGETABLES Residues of 9 organochlorine compounds were found in varying combinations in 23 of 30 composites. Organo- phosphorus residues were found in 10 of 30 composites. The most common residues and their maximum levels were DDT: 0.026 ppm; endosulfan: 0.063 ppm; para- thion: 0.012 ppm; DDE: 0.016 ppm; and dieldrin: 0.005 ppm. Cadmium (0.01-0.20 ppm) was found in 28 of 30 composites examined. Other residues found were methyl parathion. carbaryi, diazinon. 2.4-D. bromides, HCB, TDE, Dacthal (DCPA), and malathion. LEGUME VEGETABLES Residues of 5 organochlorine compounds were found in 7 of 30 composites. DDE and DDT were most com- mon, with maximum values of 0.005 ppm and 0.031 ppm, respectively. Other residues found were TDE. carbaryi, BHC, dieldrin, parathion, cadmium, and bromides. ROOT VEGETABLES Of 30 composites, 1 1 were found to contain 6 different organochlorine residues in varying combinations. The most common residues found and maximum levels de- tected were DDE: 0.006 ppm; and DDT; 0.008 ppm. Cadmium was discovered in 24 of 30 composites at 0.01-0.06 ppm. Other residues were dieldrin, TDE, o-phenylphenol, lindane, toxaphene. carbaryi, and bro- mides. GARDEN FRUITS Residues of 8 organochlorine compounds were found in varying combinations in 28 of 30 composites. The most common of these and the maximum levels were TDE; 0.087 ppm; DDT: 0.037 ppm; dieldrin: 0.010 ppm; and BHC: 0.009 ppm. Cadmium was found in 26 of 30 composites at 0.01-0.05 ppm. Other residues detected were parathion, o-phenylphenol, DDE, car- baryi, endosulfan, bromides, lindane, and Kelthane (dicofol). FRUITS Residues of 9 organochlorine compounds were found in 19 of 30 composites. TTie most common of these and the maximum levels found were DDE: 0.002 ppm; Kel- thane: 0.066 ppm; and DDT: 0.003 ppm. Residues of 5 organophosphorus compounds were found in varying combinations in 20 of 30 composites. The most com- mon and the highcEt levels found were ethion: 0.099 ppm; and malathion: 0.089 ppm. Other residues found mcluded TDE, o-phenylphenol, carbaryi, perthane, endosulfan, phosalone. lindane, diazinon, dieldrin. Botran. arsenic, cadmium, and bromides. OILS, FATS, AND SHORTENING Residues of 4 organochlorine compounds were found in varying combinations in 24 of 30 composites. The residues and their maximum levels were dieldrin: 0.006 ppm; DDE: 0.008 ppm; TDE: 0.013 ppm; and DDT: 0.012 ppm. Malathion was found in 12 composites; maximum level was 0.097 ppm. Cadmium appeared m 27 of 30 composites at levels of 0.01 to 0.04 ppm. Bromides were detected in 5 composites at a maximum of 12.0 ppm. SUGARS AND ADJUNCTS Residues of 7 organochlorine compounds were found in varying combinations in 15 of 30 composites. The most common organochlorines and their maximum levels were lindane: 0.004 ppm; DDT; 0.017 ppm; and DDE: 0.002 ppm. Heptachlor epoxide, BHC, PCP. diazinon, malathion, arsenic, and bromides were also detected. Cadmium (0.01-0.03 ppm) was found in 14 of 30 composites. BEVERAGES Of all food composites studied, beverages had the low- est amounts of pesticides; only 9 of 30 composites con- tained any residues. DDT, TDE, and lindane each ap- peared in 1 composite, cadmium appeared in 3 com- posites, and bromides appeared in 5. All residues were at or near trace levels. Discussion Organochlorine residues were found in 221 of 360 composites, or 61.4 percent of those examined. Corre- sponding percentages from previous years were 74.2 percent in 1969-70, 64.7 percent in 1968-69, and 65.6 percent in 1967-68. Organophosphorus compounds in the current reporting period were found in 77 compos- ites or 21.4 percent. The three previous reporting pe- riods showed 74, 59, and 26 composites, respectively, which had organophosphorus residues. Carbaryi was found in 20 composites during this re- porting period; 15 of these findings were at trace levels. This is in contrast to the June 1969-April 1970 period (4). which showed no carbaryi, and to the June 1968- April 1969 period (3), which showed only three positive composites. However, it should be noted that during the two previous periods the fruit and vegetable com- posites were cxamiued both before and after processing. Foods were processed in the usual manner: peeling, stripping outer leaves, cooking when appropriate, etc. This dual analysis was conducted only during the two previous reporting periods, serving as an indicator of Vol. 8. No. 2. September 1974 113 residue loss through preparation of foods for table- ready consumption. Carbaryl loss through such pro- cessing was not presented in earlier reports because of its low frequency and level of occurrence. However, carbaryl did occur as indicated in the following para- graph. Findings are presented as parts per million in both the unprocessed composite and in the diet-pre- pared composite; ND signifies no residues detected. TR signifies trace residues detected. June 1969-April 1970 Unprocessed Group VI, Legume Vege- tables, 30 composites examined Group VII, Root Vege- tables, 30 composites examined 0. 1 3 ppm Diet- prepared ND 0.14 ppm June 1968-ApriI 1969 Group VI, Legume Vege- tables, 25 composites examined 0.5 ppm TR Group IX, Fruits, 25 composites examined 0.4 ppm 0.2 ppm 0.3 ppm ND TR TR ND ND 0.3 ppm It appears that normal preparation of foods for human consumption generally renders carbaryl residues below detection levels, or very near that point. Accordingly, it is difficult to define the significance of the apparent increase in incidence during this reporting period be- cause the majority of findings were at trace levels. Analysis for o-phcnylphenol was included this report- ing period because it is detectable simultaneously with the carbaryl; o-phenylphenol was found in 10 com- posites. Arsenic residues were found in 13 composites at con- centrations from 0.1 ppm to 0.3 ppm. Of these arsenic findings, nine were in Group II: Meat, Fish, and Poultry. Cadmium was found in 213 of 360 composites; maxi- mum level was 0.20 ppm. No amitrole or dithiocarbamate residues were found during this reporting period. Chlorophenoxy acid herbi- cides (2,4-D) were found in three leafy vegetable com- posites. PCP, which is detected by the chlorophenoxy acid methodology, was reported in two composites. Pre- vious reportings showed chlorophenoxy acid herbicides in four composites in 1969-70, and four in 1968-69. Mercury residues were found in 10 of 240 composites. All mercury residues were found in Group II: Meat, Fish, and Poultry. Analyses of individual commodities within Group II have shown that the principal source of mercury in the diet is seafood. Recovery studies were conducted for all classes of chem- icals sought throughout the entire year. Table 3 lists recovery data for this reporting period for 14 of the more commonly found organochlorine residues and data for representative compounds in the other chemi- cal classes. Each recovery experiment consisted of a single determination for the unfortified food composite and a single determination for the fortified sample. Since these were performed simultaneously, the fortifi- cation level was occasionally below the level present in the sample. In other cases, not enough recoveries were run to permit statistical evaluation. These recovery data are not reported. At very low fortification levels recoveries may range from 0 to 200 percent. As the fortification level is raised, however, the recovery improves. Based on re- covery data, it is apparent that individual, low-level residues reported may vary from the so-called true value but the overall findings are useful in appraising the national residue picture. LITERATURE CITED (7) Diigi;an. R. E., and F. J. McFarland. 1967. Assess- ments include raw food and feed commodities, market basket items prepared for consumption, meat samples taken at slaughter. Pestic. Monit. J. 1(1): 1-5. (2) CorncUus.scii. P. E. 1969. Pesticide residues in total diet samples (IV). Pestic. Monit. J. 2(4) : 140-152. (3) Conulinssen. P. E. 1970. Pesticide residues in total diet samples (V). Pestic. Monit. J. 4(3):89-105. (4) Conu'liiis.sen, P. E. 1972. Pesticide residues in total diet samples (VI). Pestic. Monit. J. 5(4) : 3 13-330. (5) Diif;:^iiii. R. £., H. C. Barry, ami L. Y. Johnson. 1966. Pesticide residues in total diet samples. Science 151: 101-105. (6) Dugffan. R. E.. H. C. Barry, and L. Y. Johnson. 1967. Pesticide residues in total diet samples (lO. Pestic. Monit. J. 1(2):2-12. (71 Martin. R. J., and R. E. Duggan. 196S. Pesticide resi- dues in total diet samples (III). Pestic. Monit. J. 1(4): 1 1-20. (S) Barry, H. C, J. G. Hundley, and L. Y. Johnson. 1963. (Revised 1970.) Pesticide Analytical Manual, Vol. I and II, Food and Drug Administration, U.S. Depart- ment of Health, Education, and Welfare. (9) Hundley, H. K., and J. C. Underwood. 1970. Private communication. Also: Official Methods of Analysis, 1 1th ed.. Association of Official Analytical Chemists, Washington, D.C.. sec. 25.016. (10) Munns, R. K., and D. C. Holland. 1971. J. Ass. Offic. Anal. Chem. 54(1) :202-205. (//) Okrasinski. J. Private communication: Determination of cadmium in total diet samples. {12) Porter. M. L.. R. J. Gajan, and J. A. Burke. 1969. Acelonitrile extraction and determination of carbaryl in fruits and vegetables. J. Ass. OflRc. Anal. Chem. 52(1):177-I81. (13) Finocchiaro, J. M., and W. R. Benson. 1965. Thin layer chromatographic determination of carbaryl (Sev- in) in some foods. J. Ass. Offic. Anal. Chem. 48(4): 736-738. 14 Pesticides Monitoring Journal TABLE 2A. Levels of pesticide residues commonly found — by food class and region, June 1970-April 197 1 Chemical Baltimore Boston Kansas City Los Angeles Minneapolis I. Dairy Products ' Residues, ppm DDT Average T 0.001 0.001 T T Positive Composites Number 1 3 3 3 1 Range O.OOI T-0.005 T-0.003 T-0.002 0,001 DDE Average T T T 0.019 T Positive Composites Number 4 5 3 6 2 Range T-O.OOl T-0.002 T-O.OOl 0.010-0.028 0.001 DIELDRIN Average T 0.002 0.003 0.001 0.003 Positive Composites Number 3 5 5 6 5 Range T-0.002 0.001-0.003 T-0.007 T-0.002 0.001-0.005 BHC Average T T T T T Positive Composites Number 1 4 4 2 3 Range T T-O.OOl T-o.noi T T-O.OOl HEPTACHLOR EPOXIDE Average T T T T T Positive Composites Number 2 5 3 2 3 Range T T T-O.OOl T-O.OOl T-0.002 IL Meat, Fish, and Poultry i Residues, ppm TDE Average Positive Composites Number Range 0.003 3 T-0,009 0.003 5 T-O.OlO 0.004 4 T-0.019 T 3 T 0.001 5 T-0.002 DDT Average Positive Composites Number Range 0.005 5 T-0.015 0.009 6 T-0.018 0.013 6 T-0.033 0.005 5 T-O.OII 0.004 6 T-0.008 DDE Average Positive Composites Number Range 0.007 6 0.001-0.014 0.007 6 T-0.021 0.006 6 0.003-0.013 0.033 6 0.018-0.048 0.007 6 T-0.022 DIELDRIN Average Positive Composites Number Range 0.003 4 T-0.008 O.tlOS 4 0.001-0.015 0.005 6 0.004-0.009 0.003 6 0.001-0.004 0.003 5 0.002-0.007 PCB'S Average Positive Composites Number Range T 4 T T 2 T T 2 T T 2 T 0.029 4 T-0.15 CADMIUM Average Positive Composites Number Range 0.01 3 0.01-0.02 0.01 4 0.01-0.04 <0.01 3 0.01-0.02 0.01 2 0.01 0.01 5 0.01-0.02 ARSENIC Average Positive Composites Number Range <0.1 2 0.1-0.2 <0.1 3 0.1-0.3 <0.1 2 0.1-0.2 <0.1 2 0.2-0.3 0 HEPTACHLOR EPOXIDE Average Positive Composites Number Range T 3 T-0.003 T 5 T 0.002 3 0.001-0.011 T 2 T-O.OOl T 5 T-O.OOl MERCURY Average Positive Composites Number Range <0.02 1 <0.02 <0.02 3 T-0.03 < 0.02 1 <0.02 0.02 3 0.02-0.05 <0.02 2 0.02 (Continued next page) Vol. 8, No. 2, September 1974 115 TABLE 2A (cont'd). Levels of pesticide residues commonly found — hy food class and region, June 1970-April 1971 Chemical Baltimore Kansas City Los Angeles Minneapolis III. Grain and Cereal • Residues, ppm DDT Average T 0.001 0.002 0.001 Positive Composites Number 2 3 0 2 3 Ranpe T T-0.005 T-0.009 T-0.003 MALATHION Average 0.02.1 0.015 0.016 0.062 0.030 Positive Composites Number 6 5 5 6 6 Range 0.008-0.0.18 0.01.1-0.023 0.011-0.026 0.020-0.170 0.015-0.041 DIEIDRIN Average 0.002 T T 0 T Positive Composites Number 1 1 1 0 1 Range 0.012 0.001 T T DIAZINON Average 0.001 0 0.001 0.003 T Positive Composites Nimiber 2 0 4 4 2 Range T-0.006 T-0.006 T-0.015 T-0.002 CADMIUM Average 0.02 0.03 0.02 0.02 0.03 Positive Composites Number 5 5 5 6 6 Range 0.01-0.0-1 11.01 -0.07 0.01-0.04 0.01-0.02 0.02-0.03 PCB'S Average 0 0.06 T 0.18 0.14 Positive Composites Number 0 1 1 1 1 Range 0.36 T 0.36 0.82 BROMIDES Average 7.8 4.8 5.0 2.7 2.4 Positive Composites Number 2 2 1 2 1 Range 22-25 14 30 7.0-9.0 14 LINDANE Average 0 0 0 O.OOI T Positive Composites Number 0 0 0 2 1 Range 0.003-0.005 0,001 DDE Average T T 0 0 T Positive Composites Nimiber 2 1 0 0 3 Range T T T-O.OOl IV. Potatoes ' Residues, ppm DDT Average 0.003 0.001 0 T T Positive Composites Number 1 2 0 1 T Range 0.020 T-0.004 T 0.002 DIELDRIN Average 0.001 T 0.001 T 0.001 Positive Composites Number 1 2 3 2 2 Range 0.005 T T-0.003 T-O.OOl 0001-0.006 DDE Average 0.001 T 0.004 T T Positive Composites Number 1 t 2 1 2 Range 0.007 T 0.003-0.018 T T-0.002 CADMIUM Average 0.04 0.03 0.04 0.03 0.05 Positive Comptisiles Nlmiber 5 5 6 6 6 Range 0.02-0.08 0.02-0.04 0.02-0.08 0.02-0.04 0.02-0.09 (Continued next page) il6 Pesticides Monitoring Journal TABLE 2A (cont'd). Levels of pesticide residues commonly found — by food class and region, June 1970-April 1971 Chemical Baltimore Boston Kansas City I Los Angeles Minneapolis V. Leafy Vegetables ' Residues, ppm DDT Average 0 004 0.001 0.001 0.002 T Positive Composites Number 1 5 1 3 1 Range 0.026 T-0.004 0.008 T-0.008 0.002 DDE Average 0.003 0.003 0 0.063 0.002 Positive Composites Number 2 3 0 4 3 Range 0.004-0.016 T-0.011 T-0.008 0.002-0.006 ENDOSULFAN (I, II plus sulfate) Average 0.001 0.006 0.003 0.013 0.007 Positive Composites Number 1 4 4 4 2 Range 0.006 T-0.020 T-0.008 0.063 0.001-0.018 CADMIUM Average 0.03 0.06 0.02 0.05 0.08 Positive Composites Number 6 6 4 6 6 Range 0.01-0.05 0.02-0.14 0.02-0.03 0.03-0.08 0.02-0.20 PARATHION Average .002 0.005 T T 0 Positive Composites Number 1 4 1 1 0 Range 0.01 T-0.012 T 0.002 VI. Legume Vegetables i Residues, ppm DDT Average 0 0.006 0 T 0 Positive Composites Number 0 2 0 3 0 Range 0.003-0.031 T-0.002 DDE Average 0 0.001 0 0.001 0 Positive Composites Number 0 2 0 3 0 Range T-0.004 T-0.005 CADMIUM Average 0.007 0.007 0.007 0.003 0.003 Positive Composites Number 3 2 3 1 2 Range 0.01-0.02 0.02 0.01-0.02 0.01 0.01 VII. Root Vegetables > Residues, ppm DDT Average 0 0.001 0 T 0 Positive Composites Number 0 2 0 2 0 Range T-0.008 T-O.OOl DDE Average 0 0.001 0.001 T 0.001 Positive Composites Number 0 1 1 3 1 Range 0.005 0.006 T-0.002 0.005 CADMIUM Average 0.02 0.02 0.01 0.03 0.02 Positive Composites Number 6 5 2 5 6 Range 0.01-0.02 0.01-0.04 0.02-0.05 0.01-0.06 0.01-0.03 {Continued next page) Vol. 8, No. 2, September 1974 117 TABLE 2A (cont'd). Levels of pesticide residues commonly found — by food class and region, June 1970-April 1971 I Minneapolis I Kansas City Los Angeles VIII. Garden Fruits ^ Residues, ppm DDT Average Positive Composites Number Range 0.006 2 0.014-0.023 0.005 2 0.006-0.025 0.007 3 T-0.037 0.008 3 0.003-0.037 0.004 1 0.026 TOE Average Positive Composites Number Range 0.013 4 0.017-0.024 0.015 2 T-0.087 0.006 6 T-0.013 0.005 3 0.010-0.027 0.007 4 T-0.018 DIELDRIN Average Positive Composites Number Range 0.002 2 T-O.OlO 0.003 5 T-0.006 0.003 4 T-0.006 0.001 4 T-0.003 0.002 3 0.001-0.006 CADMIUM Average Positive Composites Number Range 0.01 5 0.01 0.01 4 0.01-0.02 0.01 6 0.01-0.04 0.02 6 0.01-0.02 0.02 5 0.01-0.05 BHC Average Positive Composites Number Range 0.002 2 0.007-0.009 T 1 0.002 T 2 T-0.003 T 1 T 0.001 0.007 IX. Fruits i Residues, ppm ETHION Average Positive Composites Number Range 0.005 2 0.009-0.02 0.004 2 0.007-0.018 0.028 5 0,008-0.073 0.004 3 T-0.015 0.017 3 0.099 KELTHANE Average Positive Coinposites Number Range 0.001 1 0.007 0.007 2 0.005-0.035 0.014 3 0.015-0.039 0.015 4 T-0.066 0.003 1 0.019 DDT Average Positive Composites Number Range 0 0 T 1 T 0 0 T 3 T-0.003 T 2 T-0.002 X. Oils, Fats, and Shortening • Residues, ppm DIELDRIN Average Positive Composites Number Range 0.001 5 T-0.003 0.001 6 T-0.006 T 4 T 0.001 2 T-0.005 T 2 T DDE Average Positive Composites Number Range 0.002 4 T-0.005 0.002 5 T-0.006 0.001 4 T-0.003 0.001 4 T-0.008 0.002 4 T-0.008 DDT Average Positive Composites Number Range 0001 4 T-0.005 0.003 5 T-0.012 0.001 3 T-0.006 0.001 4 T-0.007 T 2 T MALATHION Average Positive Composites Number Range 0.024 3 0.022-0.097 0.008 3 0.010-0.021 0.006 2 0.009-0.030 0.007 2 0.014-0.029 0.004 2 0.009-0.017 CADMIUM Average Positive Composites Number Range 0.01 5 0.01-0.02 0.01 4 0.01-0.03 0.02 6 0.01-0.03 0.02 6 0.01-0.03 0.03 6 0.02-0.04 (Continued next page) 118 Pesticides Monitoring Journal TABLE 2A (cont'd). Levels of pesticide residues commonly found — hy food class and region, June 1970-April 1971 Chemical Baltimore Boston Kansas City Los Angeles Minneapolis XI. Sugars and Adjuncts i Residues, ppm DDT Average 0 0.001 T 0.003 T Positive Composites Number 0 4 1 1 t Range T-0.004 T 0.017 T-0.003 DDE Average 0 T 0 T T Positive Composites Number 0 t 0 3 2 Range T 7-0.002 T-O.OOl LINDANE Average 0 T T 0.001 0.001 Positive Composites Number 0 2 1 2 1 Range T 0.002 0.002 T-0.004 CADMIUM Average n.oi 0.01 0.01 0.01 0.01 Positive Composites Number 2 3 3 3 3 Range 0.02 0.01-0.03 0.01-0.02 0.01-0.02 0.01 XII. Beverages i Residues, ppm BROMIDES Average 0.92 0.25 0.33 0.33 Positive Composites Number 2 0 1 1 1 Range 2.5-3.0 1.5 2.0 2.0 NOTE: denotes not applicable. T — Trace: see definition under Analytical Methods. ' Six composite samples examined from each of five districts: Baltimore, Boston. Kansas Ciiy, Los Angeles, and Minneapoli' are averages of the six composites from each site. Residues listed TABLE 2B. Pesticides found injrequenily — by food class and region, June 1970-April 1971 Pesticide District No. Composites Dairy Products i Residues, ppm HCB Boston 1 T Kansas City 1 T Los Angeles 2 n.oni. T Lindane Kansas City 2 T. T TDE Baltimore 1 T Boston 2 T, 0.001 Kansas City 3 T, T. 0.001 Los Angeles 2 T, 0 001 Minneapolis 1 T Cadmium Baltimore I 0.01 Boston 1 0.06 Los Angeles 1 0.01 Minneapolis 1 0 02 HCB Boston 1 T Kansas City 1 T Los Angeles 2 T, 0.001 Methoxychlor Kansas City 2 T, 0.008 Minneapolis 1 0,006 PCP Kansas City 1 T Bromides Baltimore 2 5.0, 5.0 Boston 1 1.5 Kansas City 1 4.0 Los Angeles 2 2.0, 5.5 Minneapolis 1 3.0 II. Meat, Fish, and Poultry Residues, ppm Lindane Boston 1 T Kansas Citv 3 T, T, O.OOl Los Angeles 1 0.001 Minneapolis 3 T, T, 0.001 BHC Boston 3 T, T, T Kansas City 2 T, 0,001 Los Angeles 2 T, T HCB Los Angeles 1 T Bromides Baltimore 2 7.5, 8.0 Boston 2 3.5, 4.5 Los Angeles 2 2.0, 6.5 Minneapolis 1 5.5 (Continued next page) Vol. 8, No. 2. September 1974 19 TABLE 2B (cont'd). Pesticides found infrequently — hy food claxs and region, June 1970-April 1971 Pesticide District | No. Composites | Amount Dieldrin Carbaryl 2.4-D Melhyl Parathion Diazjnon Bromides HCB TDE Dacihal Malalhjon III. Grain and Cereal^ Residues, ppm rv. Potatoes i Residues, ppm Boston Minneapolis Boston Baltimore Minneapolis Baltimore Boston I. OS Anpeles Baltimore Boston Los Angeles Minneapolis Baltimore Boston Kansas City Los Anyeles Minneapolis Boston Boston Minneapolis Boston Kansas City Legume Vegetables ' Residues, ppm BHC Boston Dieldrin Minneapolis TDE Baltimore Boston [ OS Angeles Minncjpolis Carbaryl 1 OS Angeles Parathion Kansas Citv Bromides Baltimore Boston Kansas City Minneapolis Heptachlor Epoxide Boston 2 T. T Carbaryl Boston 2 T. T Los Angeles T BHC Kansas City T Los Angeles T, 0.007 TDE Baltimore T Boston T Minneapolis T. 0.001, n.ooi Ronnel Kansas City 0.007 o-Phenylphenol Los Angeles T Methoxychlor Minneapolis T Botran Baltimore 0.003 Endrin Boston T Los Angeles 0.007 CIPC Los Angeles 0.14 TDE Boston T Carbaryl Boston T Kansas City T Endosulfan Boston 0 007 Los Angeles T Bromides Baltimore 2 16.0. 13.0 Boston 2 1.0. 1.0 Minneapolis 2 4.0, 2.0 Arsenic Minneapolis 1 0.1 BHC Boston 1 0.002 /. Leafy Vegetables ^ Residues, ppm 3 2 1 1 2 1 1 1 1 1 1 1 2 2 2 1 2 1 2 1 1 T, T, 0.001 0.001, 0.005 T 0.01 0.02, 0.13 T T 0.011 T 0.004 0.003 0.020 4.0, 4.0 0.5, 1.0 51.0, 3.0 2.5 2.5, 1.5 T T, 0.002 0.001 T 1 T (Continued next page) 1 T 1 T 1 T 2 T, T 3 T, o.oni, 0.003 1 0.001 1 T 1 T 2 2.0, 4.0 2 1.5, 1.5 1 5.5 1 2.0 120 Pesticides Monitoring Journal TABLE 2B (cont'd). Pcslicidcs found infreciiipntly — by food cinss and rcf^ion, June 1970-April 1971 Pesticide I District I No. Composites Amount VII. Root Vegetables ' Residues, ppm VIII. Garden Fruits ' Residues, ppm IX. Fruits i Residues, ppm Dteldrin Boston 1 o.no5 Minneapolis 1 n.ooi o-Phenylphenol Baltimore 2 T, T TDE Baltimore 2 0.007, 0.01 1 Lindane Boston 1 0.008 Los Angeles 1 0.02.1 Toxaphene Los Angeles 1 T Carbaryl Los Angeles 2 T, 0.006 Bromides Baltimore 2 5.0, 3.5 Boston 2 0.5, 6.0 Kansas City 1 1.5 Los Angeles 1 4.0 Minneapolis 1 4.0 Parathion Baltimore 2 T, 0.004 Minneapolis 1 0.005 Carbaryl Boston 2 T, 0.5 Los Angeles 1 T Endosulfan Boston 1 0.061 Los Angeles 1 T o-Phenylphenol Baltimore ) T Lindane Kansas City 1 0.002 Los Angeles 1 T Minneapolis 1 0.003 Keith ane Kansas City 1 0.019 DDE Baltimore 1 0.007 Kansas City 2 T, 0.003 Los Angeles 2 0.001. 0.002 Bromides Baltimore 2 4.5, 6.0 Boston 1 3.5 Kansas City 1 0.5 Los Angeles 2 3.0, 3.5 Miimeapolis 1 5.0 Carbaryl Baltimore 0.15 Boston T, T, 0.005 Los Angeles T, T, T Minneapolis 0.006 Perthane Boston T, T, 0.022, 0.009 Los Angeles 0.018 Botran Minneapolis 0.006 o-Phenylphenol Kansas City 0.1 Los Angeles T, T, 0.2 Minneapolis T, 0.1, 0.25 Phosalone Boston 0.278 TDE Boston T Kansas City 0.004 Los Angeles T Minneapolis 0.009 Endosulfan Boston 0.011 Los Angeles T Minneapolis T, 0.001, 0.045 Lindane Los Angeles 0.003 Diazinon Kansas City 0.009 Arsenic Baltimore 0.1 Boston 0.1 Cadmium Baltimore 0.01 Boston 0.01 Kansas City 0.03 Los Angeles 0.01 Bromides Baltimore 5.5, 6.0 Boston 0.5, 1.5 Los Angeles 1.5 Minneapolis 6.5 DDE Boston 2 T, T Kansas City 2 T, T Los Angeles 2 0.001, 0.002 Minneapolis 3 0.001, 0.001, T Dieldrin Boston 1 0.002 Los Angeles 1 T MalathioD Kansas City 1 0.036 Los Angeles 3 0.039, 0.026, 0.089 Minneapolis 3 0.021, 0.003, 0.062 (Continued next page) Vol. 8. No. 2, September 1974 121 TABLE 2B (cont'd). Pesticides found infrequently — /).v food class and region, June 1970-ApriI 1971 District No. Composites Oils, Fats, and Shortening ^ Residues, ppm Diazinon Baltimore 1 T TDE Baltimore 3 T, T, 0.013 Boston 2 o.on. 0.009 Kansas City 3 T, T, T Los Angeles 4 T. T. T. 0.007 Minneapolis 2 T, T Bromides Baltimore 2 12.0, 12.0 Boston 2 2.5, 5.5 Minneapolis 1 12.0 XI. Si:gars and Adjuncts i Residues, ppm TDE Boston T, T, 0.002 Los Angeles T Minneapolis T, 0.001 Heplachlor Epoxide Boston T Minneapolis T BHC Boston 0,002. 0.002 Los Angeles T, T Minneapolis T. 0.001 Malathion Los Angeles 0.016 Arsenic Baltimore 0.3 Bromides Baltimore 5.5. 6.0 Boston 7.5. 9.5 Kansas City 0.5, 3.5 Los Angeles 23.0 Minneapolis 12.0 PCP Kansas City 0.011 Diazinon Minneapolis T XII. Beverages ' Residues, ppm DDT Boston T TDE Boston T Lindane Los Angeles T Cadmium Baltimore 0.01 Kansas City 0.01 Los Angeles 0.01 NOTE: T = Trace: see definition under Analytical Methods 1 Six ctmipo'-ite samples examined from eacii of five districts Baliimore, Boston, Kansas City, Los Angeles, and Minnearolis. TABLE 3. Recovery experiments on pesticides found in total diet samples, June 1970-April 1971 Type of Blank Total Number of Food Spike Level,! Found.' RrCOVERY Pesticide Composite Level, ppm PPM Range PPM Range Experiments HEPTACHLOR EPOXIDE Fatty 0.005 0-0.006 (0.003) 0.008-0.010 (0.009) 2 Fatty O.IOO 0 0.097 Nonfatty 0,005 0 0-0.008 (0.005) 9 P.P-DDT Fatty 0.050 0-0.037 (0.016) 0.051-0.072 (0.059) 3 Nonfatty 0.010 0.002-0.003 (0.003) 0.010-0.012 (0.011) 3 RONNEL Fatty 0.01 0 0.008-0.010 (0.009) 2 Nonfatty 0.01 0 0 008-o.on (0.010) 3 Nonfatty 0.075 0 0.061-0.078 (0.068) 6 TDE Fatty 0.01 0-0.007 (0.003) 0.004-0.016 (0.010) 2 Nonfatty 0.01 0 0.011-0.013 (0.012) 3 DIELDRIN Fatty 0.010 0.002-0.010 (0.0061 0.010-0.020 (0.015) 2 Nonfatty 0.050 0 0.049 1 Nonfatty 0.005 0-0.008 (0.002) 0.003-0.012 (0.006) 5 ENDRIN Fatty 0.300 0 0.237 1 Fatty 0.010 0 0.008-0.010 3 Nonfatty 0.010 0 0.006-0.014 7 (0.011) (Continued next page) Pesticides Monitoring Journal TABLE 3 (cont'd). Recovery experiments on pesticides found in total diet samples, June 1970-April 1971 BHC AI DRIN CHIORDANE ^IBTI10X^•CHL0R HEPTACHLOR PCB'S TOXAPHENE MERCURY CADMIUM AKiiENIC CARBARYL u-PHENYLPHENOL Type of Food Composite Fatty Nonfatty Fatty Nonfatly Nonfally Nonfalty Fatty Nonfalty Fatty Fatly Nonfatty Fatty Nonfatty Fatly Nonfatly Fatty Nonfally Fatty Nonfalty Fatly Nonfatty Faliy Fatty Nonfatty Nonfally Fatty Fatty Nonfally Nonfally Falty Fatty Nonfatly Nonfatly Nonfatly Nonfally Nonfally Nonfally Nonfalty Nonfally Spike [,EVEL, PPM 0.005 0,003 0.003 0.050 0.005 0.005 0.005 0.10 0.20 0.1 0.1 0,2 0,2 0,1 0,1 0,003 0.003 0,075 0,075 0.200 0.200 0,10 0,02 0,1 0.02 0.1 0.04 0.10 0.04 0.5 0.1 0.5 0.15 1.00 1.0 0.5 0.20 0.5 0.2 Blank Level.' PPM Range 0-0.002 (0,001) 0-0.001 (0) 0 0 0 0 01102-0.006 (0,004) 0 0,014 0 0 0 0 0 0 0 0 0-0.017 (0.008) 0 0 0 0 0-0.024 (0.1104) 0 0-0.008 (0.001 1 O-0.O30 (0.013) 0-0.020 (0.004) 0-0.051 (0.017) 0-0.053 (0.007) 0-0.125 (0,017) 0-0,100 (0.005) 0-0.03 (0.003) 0 0-0.060 (0.005) 0 0 Total Number of Found.' Recovery PPM Range Experiments 0.003-0.007 6 (0.005) 0.003-0.004 14 (0.003) 0.002-0,003 2 (0,003) 0.041-0,043 2 (0.042) 0.004-0.007 4 (0.006) 0,003-0,008 6 (0,004) 0.008-0.010 3 (O.OOi)) 0.111-0.117 2 (0.114) 0.193 1 0.088 1 0.079-0.089 2 (0.084) 0.016-0.158 2 (0.087) 0.174-0.207 4 (0.186) 0.091-0.099 2 (0.095) 0.072-0.103 4 (0.086) 0.001-0.002 3 (0.002) 0.001-0.002 6 (0.002) 0.052-0.059 2 (0.056) 0.048-0.072 4 (0.056) 0.189-0.203 2 (0.196) 0.174-0.212 3 (0.191) 0,094-0,130 4 (0,103) 0.008-0.042 16 (0.020) 0.095-0.100 5 (0.098) 0.013-0.034 32 (0.021 ) 0.086-0,250 8 (0.123) 0.024-0.069 20 (0.036) 0.092-0.151 6 (0.116) 0.018-0.192 42 (0.047) 0.160-0.576 10 (0.439) 0.030-0,290 21 (0,116) 0.110-0.720 22 (0.412) 0.030-0,246 37 (0.127) 0.20-1.05 11 (0.701) 0.10-1.79 8 (0.98) 0.20-0.50 14 (0.48) 0.05-0.30 38 (0.18) 0.25-0.50 10 (0.33) 0-0.20 16 C0.I5) {Continued next page) Vol. 8, No. 2, September 1974 123 TABLE 3 (cont'd). Recovery experlmcnis on pesticides found in total diet samples, June 1970-April 1971 Type of Blank Total Number of Fnoi) Spike Level,! Found, I Recovery Pesticide Composite Level, ppm PPM Range PPM Range FXPERIMENTS PARATHION Falty 0.02 0 0010-0.017 (0.014) 4 Nonfalty 0.02 0 0 1)1 7-0,026 (0,021 ) 8 Nonfatty 0.10 IMI 020 (0.00.1) 0 060-0, 120 (0,09?) 6 MALATHION Fally 0.02 0-0,007 (0.002) 0.016-0.0.10 (0.022) 4 Nonfally 0.02 0-0.016 (0.002) 0.01.1-0.0.17 (0.021) 8 Nonfalty 0.10 0-O.O.tK (0.007) 0 O.Sl-0.128 (o.ion 5 DIAZINON Nonfalty 0,01 0 0,()0S-0,012 (0,010) 6 Falty 0.03 0 0,040-0,064 (0.048) 4 2,4-D Fatly 0.05 0 0 OOfi-0,040 (0,021) 7 Nonfatly nm 0 0.005-0,017 (0.021 ) 11 2,4-DB Fatly 0.0.1 0 0.0(14-0.0.16 (0.021 ) 8 Nonfalty 0 0.1 0 0.009-0.017 (0.027) 16 2,4,5-TP Fatly 0,20 0 0.0.12-0.074 (0 059) 3 Falty 0.02 0 0-0.016 (0.009) 6 Nonfatty 0.03 0 0.009-0.027 12 (0.022) ^ Numbers in pareniheses represent nveratie residue levels. 124 Pesticides Monitoring Journal GENERAL Residue Accumulation in Selected Vertebrates Following a Single Aerial Application of Mirex Bait, Louisiana — 1971-72^ H. L. Collins/ G. P. Markin,' and John Davis' ABSTRACT A survey to monitor accumulation of mirex residues in 61 species of verlebrates and certain components of the human food chain was conducted in Louisiana from May 1971 to May 1972 following a single application of mirex bait. All gas-liquid chromatographic analyses were performed on composited, whole-body homogenates. Levels of residues detected ranged from less than 0.001 to 8.4S3 ppm. > Highest concentrations of mirex were detected in logger- haul a/u/A'.v and mockingbirds 3 months after treatment. Al- though mirex was detected in 89 percent of the total samples analyzed, residues noted were usually less than 1 ppm. Introduction 'Because of increased interest in environmental effects of chlorinated pesticides, as well as increasingly refined and exacting analytical techniques, numerous papers and unpublished reports on the accumulation of mirex residues in nontarget organisms have appeared in recent years (1-5). Most of these studies have dealt with resi- dues following multiple applications of mirex bait em- ployed in attempts to eradicate or suppress isolated imported fire ant infestations. Other papers have re- ported on residues following massive doses of mirex administered to test animals in the laboratory (6-9). The majority of treated acreage now receives a single mirex application (Wa lb bait or 1.7 g actual toxicant/ acre) in State and Federal control programs aimed at providing short-term relief for landowners (10). A scarcity of information on residue levels to be expected from this type of program prompted the present study. The primary objective is to investigate and compare ' Methods Development Laboratory, Plant Protection and Quarantine Programs, Animal and Plant Health Inspection Service, U.S. Depart- ment of Agriculture, P.O. Box 989. Gulfport. Miss. 39501. ' Pest Management Program, Animal and Plant Health Inspection Service, U.S. Department of Agriculture, Mississippi State, Miss. 39762. ' Forest Science Laboratory, 3200 Jefferson Way, Forest Service, U.S. Department of Agriculture, Corvallis. Oreg. 97331. accumulation of mirex residues in selected vertebrates and certain components of the human food chain for 1 year following a single application of mirex bait. Ver- tebrates chosen represent certain families or orders whose members occupy the same general habitat, have similar food habits, and are available at all times of the year. Preliminary results of this study are reported in this paper. DESCRIPTION OF STUDY AREA The 2-square-mile study area was located in Washington Parish in the east central corner of Louisiana near the town of Bogalusa, Agricultural practices in the area are generally limited to dairy and timber farming. Per- manent pastureland and hardwood bottom land pre- dominate although hills are usually covered by stands of pine interspersed with hardwoods and underbrush. The topography is diversified enough to provide an abundance of food and cover for numerous species of vertebrates. This diversity and the fact that there had been no previous mirex treatments in the area provided the basis for its selection as a study site. The area is drained by several small streams whose flow is often interrupted during dry periods. Pushepetapa Creek, a tributary of the Pearl River, flows through the center of the study area year-around. Researchers monitored two species of fish which inhabit this body of water. Numerous farm ponds averaging less than 1 acre in size are scattered over the area and usually con- tain sizable populations of warm-water fish such as hluegills. green sunfish. and mosquito fish. Eight of these ponds were chosen as study sites; all fish samples except for those from Pushepetapa Creek were taken in these ponds. Methods and Materials BAIT APPLICATION The study area was treated with mirex bait by a Piper Pawnee aircraft on May 4 and 5, 1971, The plane was Vol, 8, No. 2, September 1974 125 equipped with a Swathmaster granular spreader cali- brated to deliver the present recommended dosage of 1.25 lb. standard 4X mirex bait per acre (1 .7 g actual toxicant/acre). Ground personnel guided the aircraft with helium-filled Kytoons. SAMPMNG PROCEDIIRFS Terrestrial animals were collected by shooting or trap- ping. Havahart live traps baited with canned sardines and peanut butter were employed to capture opossums and rodents; armadillos, birds, and rabbits were shot. A .22-caliber pistol loaded with No. 12 dust shot was used to collect frogs, snakes, and lizards. Fish were col- lected by seining or trapping with a common fish trap constructed of 1-inch mesh poultry netting and baited with cottonseed meal. These collections, while seem- ingly incomplete in some instances, represent 1.120 person-hours of work. A complete phylogenctic listing of the vertebrates sampled is presented in Table 1. Scientific names and phylogenetic arrangements general- ly follow those given by Blair et al. (11). Only the central portion of the treated area was utilized for sampling. A buffer zone one-half to one-fourth mile wide was maintained to reduce migration of study ani- mals in and out of the treated area. Some migration of the more mobile species did doubtless occur; however, the home range of most animals sampled was probably restricted to the test area. Sampling was carried out in seven excursions at 1- to .i-month intervals from April 1971 until May 1972. Unfortunately, replication of samples and thus statistical interpretation of the data were not possible because of limitations of gas-liquid chromatography CGLC) time and personnel required to replicate sampling. All in- dividuals of a given species were lumped together at TABLE 1 . Phylogenetic lislins; of selected vertebrates siimpled for mirex residues MAMMALIA Icteridae RODENTIA Sturnella ma^tm (Linnaeus)— Eastern meadowlark Cricetidae Af;elaius phoeniceus (Linnaeus )— Red-winged blackbird Reithrodontomvs humuUs (Audubon & Bachman)— Eastern Quiscalus quiscula (Linnaeus) — Common grackle harvest mouse Molothrus ater (Boddaerl) — Brown-headed cowbird Lagomorpha Thraupidae Leporidae Pyrrhitloxia cardinalis (Linnaeus) — Cardinal Syhila^tis flnridanii-: (Allen )— Eastern cottontail REPTILIA Marsupialia Squ^mata Didelrhidae Colubridae Didelphis marsupinlis (Linnaeus t^Common opossum Coluber constrictor constrictor (Linnaeus) — Southern black racer Xenarthra Natrix rhombifera rhombifera (Hallowell) — Diamond-backed Dasypodidae water snake Dasypus novemcincius (Linnaeus) — Nine-banded armadillo Natrix erythrogasler flavi^aster (Conant i—'^'ellow-bellied water snake Iguanidae AVES Anoli'i carolinensis (Voigl) — Green anole CiCONIIFORMES Sceloponi% umUdatus (Latreille) — Eastern fence lizard Ardeidae Scincidae Egretta ihula (Molina) — Snowy egret Ly^osoma laterale (Say) — Ground skink Casmerodius iilb\i\ (I innaeus) — Greater eyrel Eumeces fasciatus (Linnaeus) — Five-lined skink hydranassu tricolor (P. L. S. Muller)— I ouisjana heron Chelonia Btttorides virescens (Linnaeus )— Green heron Kinosterninae Ardeola ibis (Linnaeus) — Cattle eprel Kino\ternon subriibrum (Lacep^de) — Mud turtle Florida caeruleu (I innaeus) — I iitle blue heron Emydinae Falconiformes Terrapene Carolina (Linnaeus) — Gulf Coast box turtle Accipitridae Testudinidae Accipiter cooperii (Bonaparte) — Cooper's hawk Gopherus pohphemus ( Daudin )— Gopher tortoise Galliformes AMPHIBIA Phasianidae Salienta C'lmw. yirfiininnii\ ( Linnaeus )—Bohwhite quail Bufonidae Gallus gallus (Linnaeus) — Domestic chicken Bujo terrestris (Bonnaierre) — Southern toad Charadrhfobmes Microhylidae Charadriidae Gastrophryne carolirien'ii': (Holbrook) — Narrow-mouthed toad Charadrius vociferus (Linnaeus) — Killdeer Ranidae CUCULIFORMES Runa catesbeiana (Shaw) — Bullfrog Cuculidae Rana pipiens (Schreber) — Leopard frog Coccyzus americanus (Linnaeus )— Yellow-billed cuckoo Caudata PlCIFORMES Plethodontidae Picidae Plethodon glutinosus (Green) — Slimy salamander Melanerpes carolintis (Linnaeus) — Red-bellied woodpecker OSTEICHTHYES Melanerpes erythrocephalus (Linnaeus) — Red-headed woodpecker Cypriniformes Passeriformes Cyprinidae Tyrannidae Notemifionus crysoleucai (Mitchill) — Golden shiner Tyrannus tyraunus (Linnaeus) — Eastern kingbird Ictaluridae Corvidae Ictalurus nehulosus (LeSueur) — Brown bullhead Cyanocitfa cristata (Linnaeus) — Blue jay Poeciliidae Mimidae Gambwiia affinis (Baird & Girard)— Mosquitofish Mimus polyglolios (Linnaeusi — Mockingbird Centrarchidae Toxo^toma rttjiim (Linnaeus) — Brown thrasher Micropterus salmoides (Lacepede) — Largemoulh bass Turdidae Chaenobryttus coronarius (Bartram) — Warmouth Turdtis mif^ratoriHs (Linnaeus) — American robin Lepomis cyaneUus (Rafinesque)- Green sunfish Laniidae Lepomis megalotis (Rafinesque) — Longear sunfish Laniiis ludoviciatiHs (Linnaeus) — Loggerhead shrike Lepomis macrochirus (Rafinesque)— Bluegill 126 Pesticides Monitoring Journal each collection date and the composite was analyzed. Thus each sample usually consisted of three or more individuals except for less accessible specimens such as certain birds, opossums, and armadillos (Table 2). All samples were placed on ice immediately after cap- ture and returned to the laboratory where they were kept frozen until GLC analysis could be performed. SURVEY OF ITEMS IN HUMAN FOOD CHAIN Several recent studies have dealt with mirex residues in the human food chain. Ford et al. (12) reported on mirex residues in beef fat samples collected from cattle raised on pastures which had been treated two or more times. Others have reported on mirex residues in seafood and catfish samples (2.4,13). Certain human food items such as milk, chicken eggs. various species of fish, and game birds and animals that are occasionally consumed by man were also monitored in the present study. Milk samples were taken from a dairy located within the treated area. Eggs and domestic chickens were obtained from a flock of poultry that ranged freely over their owner's farm. METHOD OF ANALYSIS Analytical techniques sensitive to 0.001 ppm as described by Markin et al. (10) were used to determine residues in all samples. Composited samples of blended whole- body tissues were processed by grinding a 5-g portion of the blended tissue homogenate with 5 g Na^SO, in a mortar and pestle, added to 200 ml of 3 : 1 nanograde hexane and isopropanol, mixed for 1 hour on a concen- tric rotor, and placed in a separatory funnel. The solu- tion was washed three times with 200 ml distilled water to remove the isopropanol. After separation, the hexane was cleaned in ll-by-500-mm chromatographic columns packed with 10 g florisil with a 2.5-cm Na„SO| layer above and below the florisil. The hexane was concen- trated to 15 ml in a three-ball Snyder column on an explosion-proof hotplate and stored in glass-stoppered conical test tubes. Further concentration was performed when necessary by heating the extract in a water bath with an airstream filtered through a Drierite filter. Portions of the final extract measuring 1 to 7.5 iil were injected into a twin-cokmin Microtek gas chroma- tograph. Instrument parameters were: Column: (A) Glass. 1.24 m by 17 mm. racked with 3 percent DC-200 on Sureiconort 100-120 mesh. (B) Glass, 2.48 m by 17 mm, racked with a mixture of equal portions of separately coated 1.5 percent OV-17, and 1.95 percent QF-l. Detector: Electron capture using 130 MC tritium as the ionizing source. Temperatures: Injector 225°C. Columns 190°C. Detector 210°C. Carrier Gases: Purified nitrogen at 75 ml/min. Recovery averaged 97 percent when fortified samples were processed and analyzed by this procedure. Results presented have not been corrected for percent recovery. TABLE 2. Mirex residues (ppm) in selected vertebrates from pretreatment to 1 year after single mirex application Pretreatment 2 Wk 1 Mo 3 V!o 6 Mo 9 Vio 12 Mo BioTic Group and Species No. Residue No. Residue No. Residue No. Residue No. Residue No. '.ESIDUE No. Residue Analy. Found Analy. Found Analy. Found Analy. Found Analy. Found Analy. Found Analy. Found MAMMALS Eastern harvest mouse 2 0.01,1 3 0.310 1 0.450 2 0.049 2 0.014 4 0.003 1 0.010 Eastern cottontail 0 — 1 ND 1 ND 0 — 1 ND 1 0.254 0 — Opossum 1 0.120 2 0.044 0 — 1 0.099 I 0.004 0 — 0 — Armadillo 0 — 1 0.076 0 — 0 — 0 — 0 — 1 0.054 BIRDS Annual Residents Bobwhite quail 1 0.113 1 0.012 1 0.475 1 1.502 0 — 1 0.064 2 0.036 Cardinal 1 0.113 1 0.685 1 0.105 2 0.222 I 0.064 3 0.043 2 0.051 Mockingbird 1 (1.030 I 1.579 T 3.627 1 4.549 3 0.807 "> 1 .936 1 0.282 Loggerhead shrike 1 0.346 1 0.706 0 — 1 8.483 2 0.754 2 3.560 1 3.666 Blue jay I 0.007 2 0.027 2 0.035 2 0.113 2 0.181 2 0.071 1 0.198 Meadowlark 2 0.010 1 0.312 1 0.870 3 2.455 1 0.201 2 1.241 1 0.299 Killdeer 1 0.100 1 0.096 1 0.132 1 0.015 2 0.473 1 0.448 1 0.346 Brown thrasher 0 — 0 1 0.669 0 — 0 — 1 ND 1 0.624 Purple grackle 1 0,018 2 e.7.34 2 2.427 0 — 0 — 1 1.140 1 0.073 Eastern cowbird 1 0.015 0 — 1 0.713 0 — 3 0.088 3 0.028 1 0.012 Redwinged blackbird 0 — 0 — 1 0.072 1 0.280 0 — 1 0.084 2 0.208 Red-headed woodpecker 1 0.006 2 1.826 0 — 1 0.088 0 — 0 — 1 0.061 Red-bellied woodpecker 0 — 1 0.015 0 — 0 — 2 0.070 2 0.035 0 — Summer Residents Cattle egret 2 0.514 1 0.085 1 0.090 3 0.163 0 — 0 — 1 0.221 Eastern kingbird 1 0.018 1 0.121 0 — 0 — 0 — 0 — 2 0.065 Yellow-billed cuckoo 0 — 1 0.042 0 — 1 0.023 0 — 0 — 0 — Green heron 0 — 1 0.001 1 0.409 1 0.260 0 — 0 — 1 0.082 Winler Residents American robin 0 — 0 — 0 — 0 — 1 0.021 2 0.054 0 — Transients Little blue heron 0 — 0 — 1 0.101 0 — 0 — 0 — 0 — Cooper's hawk 0 — 0 — 1 0.087 0 — 0 — 0 — 0 — Snowy egret 0 — 0 — 0 — 1 0.054 0 — 0 — 0 — Louisiana heron 0 — 0 — 0 — 1 0.005 0 — 0 — 0 — Greater egret 0 — 0 — 0 — 1 0.096 0 — 0 — 0 — (Continued next page) Vol. 8, No. 2. September 1974 127 TABLE 2 (cont'd). Mirex residues (ppm) in selected vertebrates from pretreatment to I year after single mirex application Pretreatment| ! Wk 1 Mo 3 Mo 6 Mo 9 Mo 12 MO No. OF Residue No. OF Residue No. OF Residue No. OF Residue No. of Residue No. OF Residue No. OF Residue Analy. Found Analy. Found Analy. Found Analy. Found Analy. Found Analy. Found Analy. Found REPTILES Aquaiic Diamondback water snake 2 n.005 1 ND 1 0.029 0 _ 0 _ 3 0.037 1 0.054 Yellow-bellied water snake 0 — 1 0.002 0 — 0 — 1 0.078 0 — 0 — Mud turtle II — 1 (1.015 0 — 1 0.013 0 — 2 0.273 0 Terrestrial Gofer tortoise 1 o.nni 0 — 1 ND 1 0.002 0 — 0 — 1 ND Gulf coast box turtle "? ND 2 ND 1 0.009 0 — 0 — 0 — 0 — Southern black racer 1 ND 1 0.002 0 — 2 0.053 0 — 0 _ 2 o.in Southern fence lizard 0 _ 1 0.003 2 0.065 5 0.191 0 — 3 0.040 2 0.025 Ground skink 2 ND 0 — 0 — , 4 0.032 T 0.091 4 0.037 3 0.042 Five-lined skink 0 — 0 — 1 0.658 2 0.076 0 — 2 0.126 2 0.216 Green anole 2 ND I 0.011 1 0.183 0 — 3 0.072 4 0.019 3 0,017 AMPHIBIANS Slimy salamander 1 ND 1 ND 2 0.097 4 0.828 0 — 2 0.254 2 0.020 Southern toad 2 ND 2 0.001 26 0.144 3 0.030 0 _ 3 0.026 8 0.008 Leopard frog 0 — 4 0.002 1 0.015 0 — 0 — 0 — 0 — Bullfrog 0 — 1 ND 0 — 0 — 0 — 1 0.001 0 — Eastern narrow- mouthed toad 0 — 2 0.044 0 — 0 — 0 — 0 — 1 0,074 Tadpoles (undetermined species i 3 n.oi6 4 (1.005 0 — 43 0.024 0 — 0 — 0 — FISH Lenlic Habitat Largemouth bass 1 ND 1 0.018 1 0.032 1 0.624 0 — 0 _ 0 — Warmouth 4 ND 0 — 0 — 1 0.009 9 0.005 2 0.005 8 0.012 Bluegill 23 0.002 15 0.012 23 0.019 79 0.025 29 0.013 36 0,009 10 0.026 Green sunfish ->-) o.on.1 5 0.003 30 0.014 71 0.014 47 0.(H)7 36 0.002 43 0.003 Longear sunfish 1 ND 0 _ 4 0.016 4 0.039 1 0.016 0 — 0 — Golden shiner T> o.oni IQ 0.005 21 0.024 87 0.027 45 0.007 38 0.008 38 0.009 Mosquitotish 51 0.003 21 0.109 109 0.131 136 0.010 144 0,015 347 0.008 51 0.006 Brown bullhead 5 0.001 6 0.004 20 0.113 4 0.086 7 0 007 4 0.003 10 0.010 I-*nic Hahititl Bluegill 5 0.004 0 .— 2 ND 2 0.057 5 0.006 0 — 5 0.011 Longear sunfish 5 0.002 Lij 0.003 0 — 3 0.007 2 0.007 4 0.021 2 0.006 NOTE: ND = no detectable residues; — — not sampled. Results and Discussion MIREX IN SELECTED VERTEBRATES Mirex residues in selected vertebrates are shown in Table 2. With few exceptions these levels were quite low (0.001-0.005 ppm) although certain birds did accumu- late residues in the range of 1 to 8 ppm. Of particular interest are the relatively high levels detected in logger- head shrikes and mockingbirds. Authors assume that the residues reflect the diet of these birds. It is inter- esting to note that green herons did not accumulate high levels of residues as expected by virtue of their position in the aquatic food web. The extensive home range of the herons could account for their relatively low residue levels because it is possible that they de- rived the majority of their food outside the treated area. Fn general, birds such as cardinals. Eastern cowbirds. and quail, whose diets consisted primarily of seeds, fruits. and other vegetable matter, had lower levels than did the more carnivorous species such as shrikes, mocking- birds, and meadowlarks. Of the four mammal species sampled. Eastern harvest mice contained the largest amount of residues: 0.450 ppm 1 month after treatment. Reptiles and amphibians generally accumulated lower residues (0.O01-O.S28 ppm) than did birds and mam- 128 mals (0.001-8.483 ppm). Surprisingly, carnivorous reptiles such as water snakes, lizards, and skinks did not appear to concentrate mirex as did birds with similar feeding habits. This may be related to the greater vol- ume of food consumed by birds as opposed to reptiles. Birds usually consume more food per unit weight in any given time and may thus encounter and concentrate greater residues. As expected, predatory species of fish such as large- mouth bass contained more residues than did omnivor- ous species such as brown bullheads and sunfish. In general, residues detected in fish agreed fairly closely with those reported in an earlier study by Collins ei al. (N) An overview of various vertebrate trophic levels reveals that strict herbivorous species such as Eastern cotton- tails and gopher tortoises contained significantly less mirex than did either omnivorous or carnivorous species (Fig. 1 ). Regardless of trophic levels, however, residues in most animals peaked 1 to 3 months after treatment and decreased afterward. ITEMS IN HUMAN FOOD CHAIN Results of the survey of items in the human food chair are shown in Table 3. Mirex was detected in 77 per Pesticides Monitoring Journai FIGURE 1. Residue fluctuations in various vertebrate trophic levels following a single application of mircx bait cent of the samples analyzed. Detectable residues were still present in most samples 1 year after treatment. Highest concentrations found were in bobwhitc quail (0.012-1.502 ppm) and largemouth bass (0.018-0.624 ppm). Mirex was not detected in beef fat prior to or 1 year after treatment. Low levels were found in milk (0.001-0.022 ppm), chicken eggs (0.001-0.493 ppm), and chickens (0.004-0.515 ppm). These results are in general agreement with those reported by Baetcke et al (5). Their results indicated that mirex residues in milk ranged from 0.007 to 0.016 ppm; beef fat con- tained 0.012 to 0.042 ppm; chicken adipose tissue con- tained 0.087 ppm; and residues in quail ranged from 0.016 to 3.148 ppm. This and other studies have demonstrated conclusively that low levels of mirex residues remain in the environ- ment and are concentrated by some species (4.5.10,13). All residue levels reported here are well below the level reported by others as causing acute toxicity to test animals such as rats. mice, birds, and fish (7.9.15-17). However, there is virtually no information available on the long-range effects of low, chronic dosages on nontarget organisms. Future research in the area of mirex residues should not merely monitor residues; research has already shown that they exist in virtually all components of the eco- system. Instead, future research should attempt to di- vulge whether these residues are harmful, and if so, to which organisms. LITERATURE CITED (/) Butler. P. A. Monitoring pesticide pollution. 1969. Bioscience I9( 1(1) :889-89L (2) McKenzie, M. D. 1970. Fluctuations in abundance of the blue crab and factors affecting mortalities. S. C. Wildl. Resour. Div. Marine Resour. Div. Tech. Rep. No. 1. 4S pp. (.3) Lowe. J. I., P. R. Parrish. A. J. Wilson. Jr.. P. D. IVil.son. and T. W. Duke. 1971. Effects of mirex on selected estuarine organisms. Trans. 36th North Amer- ican Wildlife and Natur.il Resources Conference. Gulf Brc;ze Contrihulion No. 124. pp. 175-186. (4) Markin. G. P.. J. C. Hawthorne. H. L. Collins, and J. H. Ford. 1974. Levels of mirex and some other organochlorine residues in seafood from Atlantic and Gulf Co;istal Stales. Pestic. Monit. J. 7(3/4 ): 139-143. (5) Baetcke, K. P., J. D. Cain, and W. E. Poe. 1972. Mirex and DDT residues in wildlife and miscellaneous samples in Mis.sissippi— 1970. Pestic. Monit. J. 6(1)- 14-22. (6) Liidke. J. L., M. T. Finlay. and C. Lusk. 1971. Toxi- city of mirex to crayfish. Procambarus blandingi. Bull. Environ. Conlamin. Toxicol. 6( I ) : 89-96. (7) Van Valin, C. C. A. K. Andrews, and L. L. Eller. 1968. Some effects of mirex on two warm-water fishes. Trans-American Fish. Soc. 97(2 ): 1 85-196. (8) Naher, E. C, anil G. W. Ware. 1965. Effecls of Ke- pone and mirex on reproductive performance in the laying hen. Poul. Sci. 44:875-880. (9) Gaines, T. B. 1969. Acute toxicity of pesticides. Toxi- col. Appl. Pharmacol. 14(3 ) :5 15-534. ilO) Markin, G. P., J. H. Ford. J. C. Hawthorne. J. C. Spence. J. Davis, H. L. Collins, and C. D. Loftis. 1972. The insecticide mirex and techniques for its monitor- ing. APHIS 81-3. 19 pp. (//) Blair, W. F., A. P. Blair, P. Brodkorb. F. R. Cagle, TABLE 3. Food Item Milk Chicken Eggs Domestic Chickens Beef Fat Fishi Bluesills Brown bullheads Largemouth bass jame Animals and Birds Eastern cottontails Opossums Bobwhite quail Mirex residues (ppm) in human food chain from pretreatment to 1 year after single mirex application Pretreatment Sample Size Iqt 12 2 I lb 28 5 1 0 I 1 Residue Found 0.015 ND ND 0.003 o.oni ND n.120 0.113 2 Wk Sample Size Iqt 12 15 6 1 Residue Found ND 0.493 0.004 0.012 0.004 0.018 ND 0,044 0.012 I Mo Sample Size 1 qt 12 25 20 1 Residue Found 0.022 0.005 0.036 0.019 0.113 0.032 ND 3 Mo Sample Size Iqt ]2 0 79 4 1 0 1 1 VOTE: ND = no detectable residues; — = not sampled. Average of all samples collected from entire study area at indicated sampling in Residue Found 0.001 0.007 1 1 5 1 5 0.041 0.086 0.624 0.099 1.502 6 Mo Sample Size 1 qt 12 34 7 0 Residue Found ND ND (I 1110 0.009 0.007 ND 0.004 9 Mo Sample Size 1 qt 12 2 0 36 4 0 1 0 1 Residue Found ND 0.011 0 20] 0.009 0.003 0,254 0.064 12 Mo Sample Size Iqt 12 2 1 lb 15 10 0 0 0 2 Residue Found ND 0.001 0,014 ND 0.018 0.010 0.036 Vol. 8, No. 2, September 1974 129 and G. A. Moore. 19?:'. Verlebrates of the United States. McGrav\-Hill Book Companv. Inc., New York. N.Y. (12) Ford. J. H.. J. C. Htinlhornc. and G. P. Markin. 1971. Monitoring for niirex and other organochlorine pesti- cides in beef cattle in the Southeastern United States. Peslic. Monil. J. 7(2):S7-y4. (/.•t) Hawthorne, J. C, J. H. Ford. C. W. Collier, and G. I'. Markin. 197 J . Residues of mirex and other chlorinated pesticides in commercially raised catfish. Bull. Environ. Contam. Toxicol. (In Press) {14) Collin.t. H. L., J. R. Davi.\. and G. P. Markin. 1973. Residues of mirex in channel catfish and other aquatic organisms. Bull. Environ. Contam. Toxicol. 10(2): 73-77. (15) Gaines, T. B., and R. D. Kimbrough. 1970. Oral toxicity of mirex in adult and suckling rats. Arch. hn\iron. Hc.llh :i:7-14. (/6) Ware. G. W ., and E. E. Good. 1966. Effects of in- secticide on reproduction in the laboratory mouse. Toxicol. Appl. Pharmacol. 10:54-61. (17) Stickel, L. 1964. Wildlife Studies. Patuxtent Wildlife Rese;irch Center. Pp. 77-116. In Pesticide-Wildlife Studies, 1963. A review of fish and wildlife service investigations during the calendar year. U.S. Dept. of Interior, Fish and Wildl. Serv. Circ. 199. 1.30 Pesticides Monitoring Journal Residues of the Insecticide Mirex in Terrestrial and Aquatic Invertebrates Following a Single Aerial Application of Mirex Bait, Louisiana — 1971-72^ G. p. Miirkin," H. L. Collins,' and J. Davis' ABSTRACT Samples of 25 invertebrates were collected for I year follow- ing a broadcast application of mirex bait to 1 .000 hectares of land in southeastern Louisiana for control of the imported fire ant. Immediately following the treatment, residues of 5.504 to 22.153 ppm were found in scavengers such as the imported fire ant which fed directly upon the bait. Predatory invertebrates accumulated residues more slowly: a maximum level of 10.010 ppm was found in spiders 10 weeks after the treatment. Herbivorous invertebrates usually did not accu- mulate significant amounts of mirex. Residue levels in all terrestrial and aquatic invertebrates decreased greatly during the following year: no detectable residues were found in 68 percent of the .■•amples at the end of the .study. Introduction In 1971 an extensive monitoring program for mirex was undertaken in southeastern Louisiana. TTie purpose of the study was to determine mirex residue levels in representative groups of animals and the physical envi- ronment, and the rate of accumulation or loss of residues over a I -year period following a single treat- ment with bait for control of imported fire ants. A general description of the .study and results of verte- brate monitoring have been presented by Collins et al, '/),• results of the study of residues in the physical envi- -onment were published by Spence et al. (2). This paper 'presents data on mirex residues in invertebrate fauna. Although some information exists on mirex residues in nvertebrate populations, most of it is based on studies of animals collected from an area which received sev- Methods Development Laboratory. Animal and Plant Health Inspec- tion Service. U.S. Department of Agriculture. Box 989. Gulfport. Miss. .19501. Forest Science Laboratory, ,1200 Jefferson Way. Forest Service, U.S. Dcrartnicnl of Agriculture. Corvallis. Oreij- 97.1.11. Pest Management Program. Animal and Plant Health Inspection Service. U.S. Department of Agriculture. Mississippi State. Miss 39762. Methods Development Laboratory. Plant Protection and Quarantine Programs. Animal and Plant Health Inspection Service. U.S. De- partment of Agriculture. Gulfport, Miss. 19501. eral closely spaced treatments with bait applied to determine the feasibility of eradicating the ant from isolated areas (3,4}. .Samples were usually collected at one time and do not present any information on the rate of buildup or decline of residues in the overall population. Because mirex is annually applied to over 10 million acres (4) to control this ant, information pertaining to residues following such routine single- application control programs is more valuable in under- standing the overall residue picture for this pesticide than are data primarily concerning experimental eradi- cation programs. The present study was designed spe- cifically to observe the residue picture following the normal practice of using a single application of bait to control this ant. Sampling The treated area was a 1 .000-hectare (ha.) block of pasture and forest land in Washington Parish, La. A blanket treatment of 2.5 kg/ ha. mirex 4X bait was applied to the total area May 4 and 5. 1971. The study area and method of bait application were reported by Collins et al. (1). Researchers chose 25 groups of inver- tebrates to represent the general invertebrate fatina of the area and collected them prior to treatment and at various intervals over the following year. Initially, it was hoped that individual species could be used as indicators, but this was impractical because the brief life cycle and short season of occurrence of most inver- tebrates made it impossible to obtain most species during the entire year. It was then decided to concentrate on particular families or orders of invertebrates, the mem- bers of which occupied the same general habitat, had similar food habits, and were available at all times of the year. Aquatic samples were collected from two small farm ponds approximately one-tenth ha. in size, lying in open improved pasture, and from a 100-m section of a v'oL. 8, No. 2, September 1974 small permanent stream ("flow rate: 10 to 1.000-pliis liters/min) just above its junction with Pushepetapa Creek. The drainage system of all three bodies of water lay entirely within the treated area. From each location, bottom organisms were collected by taking 10 bottom samples in 10 to 25 cm of water. Each sample con- sisted of the bottom sediment from an area one-tenth m' and 5 cm deep. It was washed through a 20-mesh screen and the desired organisms were collected. Free- swimming organisms were collected with a hand-held sweep net. Soil insects were collected at three study sites 1 to 3 ha. in size scattered throughout the treatment area. At each site 10 soil samples, each representing an area one- tenth m-. were randomly selected; the vegetation was removed by clipping and the upper layer of duff was scraped off. Soil was excavated to a depth of 10 cm. placed on a plastic sheet, and sorted by hand to retrieve the desired animals. Surface-living arthropods were collected from the same three sites bv .^0 pitfall traps. Pitfall traps were plastic-lined paper cups 7.5 cm in diameter and 1 8 cm deep. The cup was buried with the lip level at the soil surface; each cup contained 5 cm 50 percent alcohol. Traps were left out for 24 hours and animals which had fallen into the trap were immedi- ately strained from the alcohol upon collection. Imme- diate removal of the animals from the alcohol and use of a solution which was onlv 50 percent alcohol were necessary to prevent the loss of mirex residues to the alcohol (-4). Miscellaneous animals such as web spiders, grasshop- pers, and leaf hoppers were collecled at the same three study sites bv sweeping with a sweep net 40 cm in diameter. Samples were restricted to the six study sites: three aquatic sites and the three terrestrial sites. Nests of mud dauber wasps (SccVtphron sp.) were collected from several old barns in the studv area. The six sampling sites were all located well within the treated area. A I -km buffer area was set up to minimize the chance of animal migration from the untreated area into the treated block. At the time of collection, all samples were placed in glass vials which had been prewashed in hexane and checked by gas-liquid chromatography (GLC) for contamination; alimiinum foil was used to line lids. The vials were placed on ice until they were brought to the laboratory; samples were washed in tap water, sorted, identified, weighed, and counted. Samples were deep-frozen in glass vials until processed. Collections from the three sampling sites were combined into a single composite sample for each of the 25 indicator organisms at each collection interval. Samples usually contained between 20 and 100 individuals and were usually over 1 g in weicht. However, it was sometimes necessary to use smaller samples during the winter. The smallest sample used was one-tenth g but contained at least five individual animals. A nalytical Procedures .Analytical procedures were basically the same as those used by Collins et al. (!}. who collected vertebrates from the same treatment area. Frozen samples were ground whole with a mortar and pestle with 5 g NaoSO^. added to 200 ml nanograde hexane and isopropyl alco- hol (3:1). mixed for 1 hour in a concentric rotor, and placed in a separatory funnel. Samples were washed threie times with 200 ml distilled water to remove the isopropyl alcohol. After separation, the hexane was cleaned in an ll-by-500-mm chromatographic column packed with 10 g florisil for silica gel for fish) with a 2.5-cm Na..S04 layer above and below the florisil. The hexane was concentrated in a three-hall Snvder column on an explosion-proof hotplate to 1 5 ml and was stored in glass-stoppered, conical-based test tubes. Further concentration was performed when necessary by heat- ing the hexane in a water bath with an airstream filtered through a Drierite filter. Technicians injected 1- to 7.5-ul portions of the final extract into a twin column Microtek 220 gas chromatograph. Instrument parameters were: Column: (A) Glass. 1.24 m by 17 mm. packed with T percent DC-200 on Supelcoport 100-120 mesh. (B) Glass. 2.48 m by 7 mm. packed with mixture of equal portions of separately coated 1.5 percent OV-17 and 1.95 percent OF-1 on 60-80 mesh Chromosorb. I^etector: Electron capture usinp \?0 MC Tritium as ionizing source. Temperatures Injector 225°C. Columns 190°C. Detector 210°C. Carrier Gases: Purified nitrogen at 75 ml 'min. The level of sensitivity with 1 g or larger samples was 0.001 ppm. Sensitivity of 0.1 g samples was 0.01 ppm. Recovery using this procedure averaged 97 percent when fortified samples were run through the processing and analytical procedures. Results presented have not been corrected for percent recoverv. Polychlorinated biphenyls (PCB'sl are the major con- taminants which complicate analyses for mirex (4). Particular effort was made to watch for the characteris- tic series of peaks which identify these contaminants. However, because no evidence of PCB contamination was found, no more extensive cleanup was undertaken. For a more detailed discussion of the problems of PCB"s. the types of equipment, reagents, and confirma- tion procedures used, see Markin et al. (4). Results Table 1 presents mirex residues found in 25 invertebrate groups before treatment and lor 1 year aftenvard. .Sev- eral pretreatmcnt samples contained small residues of mirex. indicating that it had been used in or near the test 132 Pesticides Monitoring Journal area before the study. However, the three aquatic and three terrestrial sites sampled were specifically known not to have a history of mirex treatment; this was one reason for their selection as sampling locations. It can be presumed then, that specimens containing residues must have migrated from adjacent fields or perhaps even from outside the treatment area. Because mirex is commonly used in Louisiana to control the fire ant, pretreatment residues were not unexpected. In general, it was felt that the small number of pretreatment samples which had mirex residues were too few to be significant in inter- preting the overall results of this study. Mirex residues were detected in all but one of the groups sampled, white fringed beetle larvae. Amounts found usually corresponded very closely to group feeding habits. General scavengers that would be attracted to oil. such as the imported fire ant, fed directly upon the bait and contained the highest mirex levels found im- mediately after the treatment. Predacious invertebrates received mirex indirectly through the food chain and therefore showed slow but progressive buildup for 20 TABLE I. Residue levels of mirex in 2? invertebrate single bait application for control of grniips saniplecl for 1 year following a the imported fire ant Pre- Mirex Residues, ppm Specimen treat- ment 24 HR 1 WK 2 UK 4 WK 10 WK 3 MO 6 MO 8 MO 10 MO 1 YR Aquatic Drafionfly Larviie Insecta: Odonata Neg n 001 0 1)05 0.005 0.011 0.013 Neg Neg Neg Neg Neg Water Scorpion Insecta: Hemiptera: Nepidae Neg 0076 0.017 0.124 0,015 0,028 0.008 0.003 0.003 0,002 0,003 Water Boatmen Insecta: Hemiptera: Corixidae Neg — 1 .003 0.146 — Neg Neg 0,016 Neg — Neg Back Swimmers Insecta: Hemiptera: Notonectidae Neg 5.504 — 0.141 0,011 — Neg 0,022 Giant Water Bugs Insecta: Hemiptera: Belostomatidae 0.001 0.038 Neg 0.013 0.038 0,004 0.004 0,009 — Neg Predacious Diving Beetles Insecta: Coleoptera: Dytiscidae 0.018 Neg 0.033 — - 0,073 — — 0,009 Neg Water Scavenger Beetles Insecta: Coleoptera: Hydrophilidae Neg 0.04.1 0.015 0,018 Neg 0005 0.011 Neg Neg Neg Neg Midge Larvae Insecta: Diptera: Chironomidae Neg 0.003 0.007 0 003 0,188 Neg Neg Neg Neg Neg Neg Crayfish Crustacea: Decapoda: Astacidae Neg 0.737 — 0.687 0,115 0,153 0.004 Neg 0.003 0,002 0.004 Leeches Hirudinea: Glossiphoniidae Neg — 0.152 0,(K13 0,004 0.006 0.011 0,007 Neg Soil Earthworms Oligochaeta: Opisthopora; Lumbricidae Neg Neg — 0.002 — 0,038 Neg Neg 0,026 0,004 0.005 Centipede Myriapoda: Chilopoda Neg Neg — 0.248 — 0,225 0.336 0.038 0.192 0,044 Millipedes Myriapoda: Diplopoda Neg Neg — Neg — 0.006 Neg — 0.076 Neg Neg White Grubs (June beetle larvae) Insecta: Coleoptera: Scarabaeidae Neg Neg — 0.005 — Neg 0.004 Neg 0.032 Neg Neg White Fringe Beetle Larvae Insecta: Coleoptera: Curculionoidea Neg Neg — Neg — — Neg Net; Neg Pitlall Traps Crickets Insecta: Orthoptera: Gryllidae Neg 1.630 — 0.072 0.040 0,030 0.003 0.009 Neg Neg Darkling Beetles Insecta: Coleoptera: Tenebrionidae Neg Neg — 0.079 0.109 0.242 0.071 0.031 0.163 0,008 Scarab Beetles Insecta: Coleoptera: Scarabaeidjie Neg 0.018 — 0.008 0.830 0.010 0.020 Neg — Neg Neg Imported Fire Ants Insecta: Hymenoptera: Formicidae Neg 22.153 — — — Neg Neg Neg — Neg Neg Wolf Spiders Arachnida: Araneida: Lycosidae Neg — — Neg 0,159 0.012 0.019 0.014 — Neg Neg Sweep Net Samples Leafhoppers Insecta: Homoptera: Cicadellidae Neg Neg — Neg 0.030 Neg — 0.018 — _ Neg Grasshoppers Insecta: Orthoptera: Acrididae Neg — — Neg 0.023 — Neg 0.008 — Neg Neg Wild Bees Insecta: Hymenoptera: Apidae Neg Neg — Neg 0.008 0,002 Neg Neg _ — Neg Crab and Web Spiders Arachnida: Araneida Neg 0.211 — 0.535 1.890 10.010 0.708 0.069 — 0.205 0,092 Other Mud daubers Insecta: Hymenoptera: Sphecidae Adults 0.008 — — — — Neg — 0.158 — — 0.076 Larvae Neg — — — — 0.524 — 0.126 — — 0.020 NOTE: All samples were adults unless otheiw All analysis was performed on a wet Neg ~ no mirex at level of detection; ise noted, whole-body basis. no sample collected. Vol. 8. No. 2, September 1974 133 to 90 days after treatment. Smallest residues were found in herbivorous animals. Most residues began to decline after 90 days and had decreased significantly in all groups by 1 year after the treatment. Detectable residues were found in only 8 of the 25 invertebrates sampled a year after treatment. LITERATURE CITED (/) Collins. H. L.. G. P. Markin. and I. Davis. 1^74. Resi- due accumulation in selected vertebrates following a (2) (3) (4) single application of mirex bait, Louisiana- -1971-72. Pestic. Monit. I. 8(21:125-130. Spencc. J. H., and G. P. Markin. 1974. Mirex residues in the physical environment following a .single bait application. 1971-72. Pestic. Monit. J. 8( 2 ): 135-139. Bactcke, K. P.. J. D. Cain, and W. E. Poe. 1972. Mirex and DDT residues in wildlife and miscellaneous samples in Mississippi, 1970. Pestic. Monit. J. 6(1): 14-22. Markin, G. P., J. H. Ford, J. C. Hawthorne. J. H. Spence. J. Davis, H. L. Collins, and C. D. Loflis. 1972. The insecticide mirex and techniques for its monitoring. USDA, APHIS 81-3, Nov. 1972, 19 pp. 134 Pesticides Monitoring Journal Mirex Residues in the Physical Environment Following a Single Bait Application, 1971-72 James H. Spence ' and George P. Markin " ABSTRACT In 1071 and 1972. .san}ph:<: of soil, scdinunt. water, and Bahiii grass were collected at various intervals in Louisiana and Mississippi after the areas had been aerially treated with mircx to control the red imported fire ant (Solenopsis invicta Biiren). In Louisiana, samples were collected through- out the first year after spraying: in Mississippi, iliev were taken for the first 4 months. Samples had also been collected from the physical environment in both States before spraying began. Residues in the sediment at the bottom of ponds increased graduallv after treatment, indi- cating that mire.x was being carried in />v runoff water. Residues reached maximum levels 10.7 and I.I ppbl about I month after treatnient, and graduallx declined thereafter. Soil residues in the Louisiana study built up to a peak of 2.5 ppb after I month and gradually declined over the remainder of the year. Bahia grass in Louisiana had residues in both roots and blades, indicating that some translocation occurred from the roots upward. In Mississippi, residue patterns of pond sediment were less predictable than in sediment of Louisiana ponds, but levels peaked at about the same interval: I month after treatment. Water in the pond .showed the highest residue levels imme- dialelr after treatment. 0.020 and O.y^l pnh. and still had detectable levels of 0.001 to 0.005 ppb as long as 3 months after treatment. Bahia grass roots and blades were combined for analysis; the highest residue levels (26 ppb) occurred between 3 and 4 months after treatment. hurodiiction Mirex is presently the major chemical used to control the red imported fire ant {Solenopsis invicta Biiren: for- merly referred to as Solenopsis saevissima richteri Forel). Information about the enxironniental impact ' Animal and Plant Health Inspection Service, US, Deparimcnt of Agriculture, P.O. Box 989, Gulfport, Miss, .195UI, ' Forest Science Laboratory, 3200 JefTerson Way, Forest Service, U.S. Department of Agriculture, Corvailis, Oreg. 973.^1. of mirex in bait form is very limited. Although some of the mirex that is applied as bait soon enters the biomass of an area, the degradation pattern of the remaining mirex. which is presumbaly in the physical portion of the environment, has yet to be determined. In 1971 an in-depth study was undertaken to determine the movement and degradation pattern of mirex in an area where it had been applied to control the imported fire ant. Mirex residue levels occurring in various indi- cator species or groups of animals in that study have been presented in two previous papers (1.2). This paper reports levels of mirex residues occurring in portions of the physical environment, soil, sediment, and water at various intervals up to 1 year after treatment. Also in- cluded in this report are the residue levels found in Bahia grass iPaspaluin iiotalitiu Fltiggc). the most abun- dant pasture grass in the area and the only plant moni- tored during the study. This study was set up in Washington Parish. La. Un- fortunately, it was determined that the system used for monitoring water was not satisfactory after the study had been under way for several months. Therefore, data for water from the first study site were discarded. In February 1972 a second study was set up in Harrison County. Miss., which concentrated on mirex levels in water. The newer method of sampling was not totally satisfactory but was believed to present data accurate enough for publication in this paper. .Study Area The first study ran for 1 year in Washington Parish. La., on a 2.00()-acre area of land that is farmed, grazed, or forested. The open area of the treated block was pre- dominantly Bahia grass, hay fields, or pastures. Most of the pastures, however, contained small artificial ponds used for stock watering, ,^uthors chose three separate Vol.. 8. No, 2, Slpiemufk 1974 135 pastures, each with its own pond, as sampling sites. Study sites were the same as those used by Markin et al. (2) to collect invertebrates and were in the same general area where many of the vertebrates reported by Collins et al. (/) had been collected. The second study site was a 100-acre block of cutover pine forest and Bahia grass pastures located east of Saucier, Miss., in the northern part of Harrison County. The center of this plot was dominated by a 2-acre pond, the entire drainage system of which lay within the treated area. Water in the pond was sampled at both the surface and at its deepest point, approximately 1.5 m. Both study sites were aerially treated with one applica- tion of standard 4X mirex bait (1.25 lb bait containing 1.74 g actual mirex/acre). The Louisiana block was treated May 4 and 5. 1971; the Mississippi block was treated February 15, 1972. Sampling Procedures Sediment samples were collected with a hand-thrown grab sampler at 10 locations around the ponds. Samples were combined at the site and a subsample was removed for analysis in the laboratory. Soil was collected from 10 random sites, each 0.1 m-. where vegetation had been clipped and litter brushed away. Soil was removed to a depth of 7.5 cm. The 10 samples were combined on the field into a composite sample and a subsample was removed and brought to the laboratory. Grass was clipped from these areas and set aside. Dirt was shaken from the roots which were also sampled. In the Louisiana study, stems and leaves of grass were saved as one sample; roots were a separate sample. Residues found in biological indicators that were col- lected have been presented elsewhere (1.2). In the Mississippi study, biological indicators were not col- lected because the main emphasis was on determining mirex residues in water. For a more detailed description of the collection techniques used, see Markin et al. (3). In both Louisiana and Misissippi. sampling varied ac- cording to weather conditions and accessibility of the sampling sites, A sample was taken from both sites before treatment; the next sample was obtained 1-3 hours later, SubseqLient collections were made 24, 48, and 72 hours after treatment; then sampling continued at various intervals for a given period of time: 4 months in Louisiana and 1 ye;ir in Mississippi. Water was sampled with a pair of filters. The first filter, a Sears sediment filter containing a replaceable cellulose cartridge, removed suspended particulate mat- ter. The second filter, a Sears taste/odor filter contain- ing a replaceable activated charcoal cartridge, removed dissolved material. Water was pumped through the filters using a 115-volt submergible pump. Rate of flow varied from 25 gallons an hour at the start of a sampling period to less than 1 gallon an hour 24 hours later. Variation in flow was caused by clogging in the sediment filter which automatically reduced flow rate. To determine the total volume of water sampled during each period, three 55-gallon drums were set up in series below the filtering system to collect the filtered water so that it could be measured. Water was collected at two locations in the pond, the pond surface and the bottom at the deepest point, 1.5 m. Surface water was taken by floating the pump in a styro- foam boat so that the pump inlet was just under the pond surface. Bottom water was sampled as water emerged from a drainage pipe under the dam. Re- searchers used two pumps and two separate filter sys- tems; each had its own pump and collection barrels. A nalytical Procedures To begin soil extraction, a 5-galIon cream can was partly filled with 5 kg moist soil, and sufficient 3:1 acetone :hexane to cover the soil to a depth of 3 cm. After 2 hours of rotation, the extract was poured through prewashed fluted filter paper into a separatory funnel. The acetone was washed out with water and the remaining hexane extract was dried over anhydrous sodiimi sulfate. The extract was treated with concen- trated sulfuric acid until it was clear and colorless; then it was injected into the gas chromatograph. Sediment was extracted similarly except that 10-kg samples were used and the water that rose from the sediment while standing was extracted separately. Total mirex in this sample was added to mirex from the sediment sample. The Bahia grass was ground in a Waring blender with a small amount of water, extracted with acetone, then processed as the soil and sediment had been. For a detailed description of the analytical procedures above, see Markin et el, i3). The wet cellulose cartridge used to extract water was soaked with acetone, allowed to drain into a separatory funnel, soaked with methylene chloride, allowed to drain, and then soaked again with acetone. After drain- ing was completed, the acetone was washed out with water, the bottom layer was dried over sodiimi sulfate, and the extract was concentrated into hexane using a Snyder colimin. The sample was then treated with con- centrated sulfuric acid and analyzed without further concentration. The wet charcoal from the activated charcoal filter was removed from the plastic housing and subjected to Soxhlet extraction with methylene chloride for 3 hours. After drying, the extract was concentrated into hexane and treated with concentrated sulfuric acid. The ex- tract was then ready for analysis without further con- centration. After completion of the study it was deter- mined that extracting the activated charcoal could have been improved by multiple extractions using a wet 136 Pesticides Monitoring Journal solvent such as water-saturated benzene to remove more niirex. This method js described by Thornburg (4). Primary indentification and quantification of mirex was accomphshed on a Hewlett-Packard 402 gas chroma- tograph. Researchers employed two columns with differ- ent characteristics. Instrument parameters were: Columns; (AlCrkiss. 1 ft bv ' : in,, racked wilh ^ ncrcenl DC- 200 on 100/120 mesh Supelcoport. IB) Glass. .1 fi hy ' .. in., necked wilh a mixture of equal portions of separately coated ?l percent DC- 200 on 100/120 mesh Supelcoport and 5 percent XE-60 on 60 SO mesh Chromosorb W, Detectors: Electron capture, with 1.10 MC tritium ionizine source. Temperatures: Injector Z.'S'C Column 190°C. Deteclor 2I()°C. Carrier Gas: Prepurified nitrogen flowing at 90 ml/min (Column A) and 36 ml/mm (Column B). Purge Gas: Methane: 5 percent: argon: 95 percent. Confirnuition ami Recovery Because residue levels were low, confirmation was limited to extraction p-values and retention time com- parisons of spiked samples to a mirex standard. The typical responses of PCB's were not observed in any sample; presumably no PCB's were present. Recovery rates were established for the samples by spiking them with a known concentration of mirex at approximately the same level as mirex found in the samples. Re- coveries were: sediment, 95 percent; soil, 97 percent; and grass. 97 percent. Tabular values for these samples have been corrected for recovery. The rate of recovery for filtered water was 70 percent, but the recovery method was not considered valid; thus tabular values for filtered water have not been corrected for recovery. Sensitivity for soil, sediment, and grass was 0.01 ppb; it was 0.001 ppb for water. Results and Discussion Pretreatment samples of sediment and grass showed detectable mirex residues. Although the land had never received aerial applications of bait, some landowners in both treated areas were known to have used mirex on an individual basis. TABLE 1. Mirex residues in sediment collected in Mississippi (1972) and Louisiana (1971-72) Days POSlTRI;ATMENT MlHEX, PPB Mississippi LoUtSIANA' Cumulative Rainfall, in. Pretreatment 0.06 0.0 1-3 hours 0.16 0.0 I 0.41 0.0 2 0.17 0.0 3 0.51 0.0 7 0.11 0.3 14 O.ll 0.4 21 0.30 2.4 28 0.93 4.1 35 1.1 5.3 42 0.85 6.0 49 0.04 7.1 56 0.03 7.6 63 0.20 7.6 70 0.01 7.6 77 0.02 9.5 84 0.05 11.3 91 0.10 16.1 98 0.01 19.5 105 0.76 19.5 Pretreatment Neg 1-3 hours Neg 3 Neg 4 Neg 7 0.02 17 0.04 25 0.30 31 0.70 38 0.60 45 0.03 54 0.08 69 0.10 77 0.10 90 0.60 117 0.30 164 0.20 185 0.09 223 0.06 268 0.50 337 0.16 374 0.40 ■lOTE: Neg = no mirej at level of detection. Louisiana ramfall not recorded. /OL. 8, No. 2, S fcPTEMBER 1974 TABLE 2. Mirex residues in soil collected in Mississippi (1972) and Louisiana (1971-72) Days posttreatment Mirex, ppb Mississippi Pretreatment _ 1-3 hours 6.3 1 0.71 2 0.31 3 4.2 7 0.87 14 0.41 21 0.22 28 1.2 35 1.3 42 4.0 49 0.78 56 2.7 63 8.2 70 1.5 77 1.2 84 2.0 91 10.4 98 2.4 105 1.1 112 0.31 Louisiana Pretreatment _ 1-3 hours 0.70 3 0.70 4 0.30 7 0.80 17 0.70 25 2.0 31 2.0 38 2.5 45 2.5 54 2.5 69 1.4 77 1.4 90 1.0 117 1.0 164 0.60 185 0.90 223 0.90 268 0.20 337 0.20 374 — NOTE: — = no d ata. 137 Mirex residues in sediment collected from the bottom of the study pond in Louisiana (1971-72) and Mississippi (1972) are shown in Table 1. Rainfall records were kept for the duration of the Mississippi studies. Mirex residues in soil collected from the two study sites are shown in Table 2. Given the rate of application, 1.7 g of actual mirex per acre, the expected residue level for mirex in the upper 7.5 cm of the soil was 3.76 ppb. In Louisiana the residue level foimd immediately after treatment was much below this and possibly indicates that a foraging insect such as the fire ant had collected much of the material from the soil surface. Buildup of soil residues in Louisiana over the next 54 days has been interpreted as an indication that the mirex was slowly returning to the soil through decomposition of dead insects or excretion. The decline in residues after 54 days could be caused by either degradation of mirex or translocation out of the soil. Residue levels found in soil in Mississippi are much more variable and show no distinctive pattern. A possible explanation is that bait was applied during winter months when animal activity was minimal. Mirex residues in Bahia grass from Mississippi and Louisiana are shown in Table 3. In the Louisiana study where grass was not sampled until 69 days after treat- ment, the blades and roots were analyzed separately to determine mirex distribution in the Bahia grass. Mirex findings in the above-ground parts of the grass, even after thorough cleaning to remove all external dirt, indicate that some translocation was taking place from the roots to the blades. This agrees with Mehen- dale et al. (5), who foimd an uptake and translocation of mirex by beans and pea seedlings in laboratory tests. In Mississippi the roots and blades were combined for analysis. Table 4 shows mirex residues in water collected from the two sites in the Mississippi pond, using the double filter systems. Water samples from the bottom of the pond show residue values which remain higher and TABLE 3. Mirex residues in Bahia grass (Paspalum notauim Fliiv.vel collected in Mississippi {1972) and Louisiana (1971-72) Days posttreatment Blades, ppb Roots, ppb Louisiana 69 17.0 77 2.0 90 1.6 117 4.0 164 6.5 185 3.6 223 2.6 268 5.7 337 6.0 374 0.01 9.0 1.0 0.3 0.6 2.7 0.8 2.3 4.9 17.0 0.07 Blades and roots, ppb Pretreatment 0.14 1-3 hours — — 6.0 1 — — 2.7 2 — — 4.4 3 — — 1.8 7 — 4.0 14 — — 9.0 21 — — 1.8 28 — — 3.2 35 — — 1.7 42 — — 3.7 49 — — 14.0 56 — _ 10.0 63 — — 4.7 70 — — 16.0 77 — — 12.0 84 — — 3.9 91 — 16.0 98 — — 13.0 105 — — 26.0 112 — — 3.1 26.0 3.0 1.9 4.6 9.2 4.4 4.9 106 23.0 0.08 NOTE: ; no data. TABLE 4. Mire.x residues in fillered water collected in Mississippi, 1972 Filter System 1 Filter System 2 Days posttreatment (deepest point) (pond surface) Cartridge: Cartmdoe: Cartridoe: Activated Total Cartridge: Activated Total Sediment, ppb Carbon, ppb Mirex, ppb Sediment, ppb Carbon, ppb Mirex, ppb Pretreatment Neg Neg Neg Neg Neg Neg 1-3 hours 0.033 Neg 0.033 0.007 0.009 0.016 1 0.530 Neg 0.530 0.008 0.012 0.020 2 0.001 Neg 0.00 1 0.003 0.004 0.007 3 0.001 0.006 0,007 0.001 0.002 0.003 7 0.001 0.007 0,008 0.005 0.001 0.006 14 0.003 0.010 0,013 0.006 0.001 0.007 21 0.002 0.012 0,014 0.002 0.003 0.005 28 Net: 0.010 0,010 0.001 0,002 0.003 35 0.001 0,012 0,012 Neg 0.002 0.002 42 Ncf; 0.003 0.003 Neg 0.001 0.001 49 0.004 0.004 0.008 Nee 0.002 0.002 36 0.002 0.003 0.005 Neg Neg Neg 63 0,001 0,009 0.010 0.001 0.001 0.002 70 0.001 0.010 0.011 0,001 0,001 0.001 77 0,0(11 0,005 0,006 0,001 0,001 0.001 84 0,001 0.010 0,011 0.002 Neg 0,002 91 0.002 0.003 0,005 0.002 0.001 0.003 98 0.001 0.015 0.016 0.003 Neg 0,003 105 o.noi 0,003 0.004 0,007 0.003 0,010 112 0.001 0,002 0.003 0,005 0.002 0.007 NOTE: Neg = no mirex at level of detection. 138 Pesticidhs Monitoring Journai more constant than those from the surface of the pond. Because sample water flowed through both the sediment and the activated charcoal filters, the total mirex residue load for each sampling period is determined by adding the figures for the two different filters. In general, highest residue levels were found immediately after treatment. Acknowledgment For assistance and suggestions, authors are indebted to Dudley J. Adams. Animal and Plant Health Inspection Service, U.S. Department of Agriculture, Gulfport, Miss. LITERATURE CITED (/) Collins, H. L., G. P. Markin, and J. Davis. 1974. Res- idue accumuhition in selected vertebrates following a single aerial application of mirex bait, Louisiana — 1971- 72. Pestic. Monit. J. 8(2) : 125-130. (2) Markin, G. P.. H. L. Collins, and J. Davis. 1974. Resi- dues of the insecticide mirex in terrestrial and aquatic invertebrates following a single aerial application of mirex bait, Louisiana — 1971-72. Pestic. Monit. J. 8(2): 131-1.^4. (i) Markin, G. P., J. H. Ford, J. C. Hawthorne, J. H. Spence, J. Davis, H. L. Collins, and C. D. Loflis. 1972. The insecticide mirex and techniques for its monitoring. APHIS 81-83 U.S. Department of Agriculture, Animal and Plant Health Inspection Service. 19 pp. (4) Thornburg, W . W. 1972. Analytical methods for pesti- cides, plant growth regulators and food additives. Vol. 1. Academic Press, New York and London, p. 97. (5) Mehendale, H. M., L. Fishbein, M. Fields, and H. B. Matthews. 1972. Fate of mirex-c" in the rat and plants. Bull. Environ. Conlam. Toxicol. 8(4) :20O-207. Vol. 8, No. 2, September 1974 139 BRIEFS Organochlorine Insecticide Residues in Carpeting ^ David L. Mick,= Herbert Hetzler," and Edwin Slach ' ABSTRACT Organochlorine insecticide (OCI) residues were detected by tuis-licjiiid chromatography in carpet samples in 1973. Diel- drin was found in largest quantities: it was also found most frequently, existing in 84 percent of the samples. Wool carpet had higher residue levels than did synthetic carpet. OCI residues were also found in raw wool, but at much lower levels, indicating that most of the residues had been absorbed at the carpet mill rather than on the farm. Introduction Insect damage to carpeting can be prevented at carpet mills by treating fibers with insecticides during the man- ufacturing process. Garrison and Hill found dieldrin residues while monitoring the aqueous effluent of a mill (!}. Their research stimulated this study to determine organochlorine insecticide residue levels in carpeting. The authors wished to explore carpeting as a possible source of insecticide residues in organic samples, par- ticularly human tissue, which have no apparent ex- posure to chlorinated hydrocarbons. Saniplint; and .Analytical Procedures Carpet remnants were obtained from carpet retailers and homes around Iowa City, Iowa, in 1973. Selection was not based upon any specific manufacturer, fiber, or color. No chlorinated hvdrocarbon insecticides had been applied in the vicinity of the carpets while in the store or home. Researchers extracted 1 0-g samples of one-inch carpet squares by immersing them in 5()()-ml, glass-stoppered, Erlenmeyer flasks with 200 ml n-hexane for 15 minutes. 1 Iowa Community Pesticides Study. Institute of Agricultural Medi- cine. College of Medicine. University of Iowa. (Reprints are avail- able from this address.) Work was performed pursuant to U.S. Environmental Protection Agency — Office of Pesticides Programs. Technical Services Division Contract No. 68-02-0746. 3 Iowa Department of Environmental Quality, 3920 Delaware Avenue, Des Moines, Iowa 50316. hand-shaking the flasks for 5 minutes, and filtering the solutions through Whatman No. 1 filter paper. The n-hexane extract was concentrated by flash evaporation to approximately 5 ml prior to cleanup on a florisil column. The column was first eluted with 200 ml 6 percent diethyl ether in petroleum ether and then with 200 ml 15 percent diethyl ether in petroleum ether. Eluates were concentrated to approximately 5 ml and subsequently analyzed by gas-liquid chromatography using QF-l/SF-30 and DEGS columns. Sensitivity was approximately 0.001 ppm for aldrin, dieldrin, lindane, and p./j'-DDE, and 0.002 ppm for p.p'-DDT. Thin- layer chromatography was used to confirm detected residues. Residts and Discussion Every carpet sampled revealed residues of at least one chlorinated hydrocarbon: aldrin, DDE. DDT, dieldrin, or lindane (Table 1). DDT and dieldrin were found most frequently, occurring in 15 and 21 of the 25 samples, respectively; Dieldrin was found in consider- i ably higher quantities than any other insecticide. High- est residue levels of DDT and dieldrin appeared in wool carpeting. These levels probably result from treatment at the mill rather than on the farm, although the current study produces only limited evidence to support this conclusion. To establish a control for wool samples, raw wool was obtained from a central Iowa farm and processed as the carpet samples had been. Although wool was sam- pled from only one farm (25-30 sheep), authors con- sider it representative of raw wool throughout the West and Midwest which is used in the manufacture of car- peting. Chlorinated hydrocarbon insecticides detected in a 6.7-g sample of raw wool are shown in Table 2. The fact that the raw wool contained residues indicated that the environment of sheep contributes to insecticide residue levels found in wool carpet samples. But these 140 Pesticides Monitoring Journal TABLE 1. Chlorinated hydrocarbon insecticide residues in carpeting, Iowa City, Iowa — 1973 Residue Levels, ppm Fiber Aldrin Dieldrin P,p'-DDT p,p'-DDE Lindane Wool 0 27,6.18 0.638 0.036 0 ** 1) 9.684 0.424 0.159 0 " 0.105 9,3.11 0.234 0 0 " 0,022 7,391 0.030 0 0 Synthetic 0 25,393 0.142 0 0 0.070 2.383 0 0 0 0 0.956 0.240 0.032 0 0.011 0.825 0 0 0 0 0.497 0.028 0 0 0 0.361 0 0 0 0 0.119 0.082 0 0 0 0.116 0.095 0 0 0 0.075 0 0 0 0.026 0.071 0 0 0 0 0.040 0.021 0 0 0 0.026 0.051 0 0 0 0.019 0 0 0 0 0.010 0 0 0 0.010 0.008 0 0 0.007 0 0.007 0 0 0 0 0.004 0 0 0 0.041 0 0.063 0 0.015 0.049 0 0.086 0 0 1) 0 0.025 0 0 U 0 0.863 0 0 TABLE 2. Chlorinated hydrocarbon insecticide residues in raw wool, Iowa — 1973 Insecticide Residue Levels, ppm P,p'-DDT P,P-DDE o.p'-DDT Dieldrin 0.064 0.009 0.015 0.017 levels were not great enough to account for all residues detected in the samples. Recovery studies indicate that the extraction process detected 85 to 100 percent of the residues in both wool and synthetic carpet. This is based upon the difference in insecticide residues recovered from nonspiked sam- ples contrasted to those recovered from spiked samples. As postulated earlier, results of this study provide evi- dence for a potential source of human exposure to chlorinated hydrocarbon insecticides. Findings may help explain why insecticide residue levels are sometimes detected in organic samples, such as human tissue, with no apparent source of exposure. LITERATURE CITED (/) Garrison. A. W., and D. W. Hill. 1972. Organic pol- lutants from mill persist in downstream waters. Amer. Dyest. Rep. 61(2):21-23. Vol. 8. No. 2, September 1974 141 Organochlorine Pesticide Residue Levels in North American Timber Wolves — 1969-71 James C. Schneeweis,' Yvonne A. Greichus," and Raymond L. Linder ' ABSTRACT Tongue and mii.ule tissue samples from 51 limber wolves captured in Minnesota and northwest Ontario in 1969-71 were analyzed for ori;anochlorine pesticide residues in an attempt to determine the level of insecticide contamination in their environment. No determined residue levels ex- ceeded the minimum analytical confidence limits established in a previous study by one of the authors. The implementa- tion of new Slate, national, and international leaislalion pro- tectinfi timber wolves makes this study especially valuable because samples from a wide range of age groups will no longer be readily available for analysis. Introduction The timber wolf {Canis lupus) has recently become a sym- bol of wilderness environments. Because these mammals are at the top of their food chain, one would expect their tissues to contain pesticide residues if their environment were contaminated by organochlorine compounds. The pur- pose of this paper is to report results of analyses of timber wolf tissues for selected pesticides. This study is especially valuable because recent State, national, and international legislation protecting timber wolves has made samples from a wide variety of age groups more difficult to obtain for analysis. Analytical Methods Tongue and muscle tissue samples were taken from 51 timber wolves captured by hunters and trappers: 30 from ^ Minnesota Department of Natural Resources. International Falls, Minn. 56649. ■ Experiment Station Biochemistry Department. Souttl Dakota State University, Brookinys, S. Dak. ^ South Dakota Cooperative Wildlife Research Unit, Brookings, S. Dak. the winter of 1969-70 and 21 from 1970-71. No animals were collected specifically for this study. Wolves were assigned an age class based on examination of the gross size and weight of the carcass, wear and development of the teeth, and, in some cases, development of the cranium (Table 1). Wolves were from Koochiching and northwest St. Louis Counties, Minn., and from northwest Ontario, Canada. TTie area is wild to semiwild in nature and is primarily coniferous forest with little or no human dis- turbance. Samples were kept frozen until they were analyzed in the Experiment Station Biochemistry Laboratory, South Dakota State University. Analytical procedures were those reported by Greichus, et al. {1,2). Results No determined residtie levels exceeded the minimum ana- lytical confidence limits establishd by Greichus in 1973 (2). These limits are: lindane, heptachlor. heptachlor epoxide, aldrin, dieldrin, and DDE — 0.01 parts per million (ppm); DDT and DDD— 0.05 ppm: and PCB's, endrin, and meth- oxychlor — 1.00 ppm. These data suggest that pesticide contamination was quite low in specified areas of Minnesota and Ontario. LITERATURE CITED (/) Greichus. Yvonne, D. Lamb, and C. Garrett. 1968. Efficiency of extraction of metabolically incorporated HEOD (Carbon- 14) from pheasant tissues, eggs and feces. Analyst 93:323-325. (2) Greichus. Yvonne A., A. Greichus. and R. J. Emerick. 1973. Insecticides, polychlorinated biphenyls and mer- cury in wild cormorants, pelicans, their eggs, food and environment. Bull. Environ. Contam. Toxicol. 9(6): 321-328. 142 Pesticides Monitoring Journal TABLE 1. Sex and age class ^ of 51 North American timber wolves analyzed for insecticide residues, 1969-71 Number Analyzed Males Females Age Age Year 2 Adults Juveniles Unknown Adults Juveniles Unknown Total 1969-70 12 0 2 5 5 6 30 1970-71 ND ND 8 ND ND 13 21 Total 12 0 10 5 5 19 51 NOTE: ND — no data. I Juveniles were those born around the preceding spring. ' Samples were obtained both winters between December and March. Vol. 8, No. 2, September 1974 143 APPENDIX Chemical Names of Compounds Discussed in This Issue ^ ALDRIN AMIBEN AMITROLE ATRAZINE BHC (BENZENE HEXACHLORIDE) BIDRIN 8 BOTRAN 9 CAFTAN CARBARYL CHLORDANE CHLORPROPHAM CIPC COTORAN ® 2,4-D DACTHAL « DCPA DDD DDE DDT DEF» DIAZINON DICHLORAN DICOFOL DICROTOPHOS DIELDRIN DIURON ENDOSULFAN ENDRIN FLUOMETURON FOLEX HCB HEPTACHLOR HEPTACHLOR EPOXIDE KELTHANE ® LINDANE MALATHION METHOXYCHLOR METHYL PARATHION MIREX MSMA PARATHION PCB's (POLYCHLOR- INATED BIPHENYLS) PCP PERTHANE o-PHENYLPHENOL PHOSALONE PROPACHLOR RAMROD® RONNEL TDE TOXAPHENE TRIFLURALIN Not less than 95% of 1, 2,3.4,10, 10-HexachIoro-l, 4,4a. 5, 8, 8a-hexahydro-l,4-endo-ejco-5,8-diniethaiionaphthalene 3-Amino-2,5-dichlorobenzoic acid 3-Amino-l,2.4-triazole 2-Chloro-4-ethylammo-6-isopropylammo-j-triazine 1,2.3.4.5.6-Hexachlorocyclohexane (mixture of isomers). Commercial product contains several isomers of which gamma is most active as an insecticide. See dicrotophos. See dichloran. N-Trichloromethylthio-4-cyclohexane-l,2-dicarboximide 1-Naphthyl ^-methylcarbamate 1.2.3,4,5,6,7,8,8-Octachloro-2.3,3a,4,7,7a-hexahydro-4.7-methanoindane. The technical product is a mixture of several compounds including heptachlor, chlordene, and two isomeric forms of chlordane. Isopropyl JV-(3-chlorophenyl) carbamate See chlorpropham. See fluometuron. 2,4-Dichlorophenoxyacetic acid See DCPA. Dimethyl 2,3.5.6-tetrachloroterephthaIate See TDE. Dichlorodiphenyl dichloro-ethylene (degradation product of DDT). Main component (p.p'-DDE): !.l-Dichloro-2.2-bis(p-chlorophenyl) ethylene o,p'-DDE: l,l-Dichloro-2-(o-chlorophenyl )-2-(p-chlorophenyl) ethylene Main component (p,p'-DDT): a-Bis(p-chlorophenyl)^.^.|3-trichIoroethane Other isomers are possible and some are present in the commercial product. o,p'-DDT: [l,l,l-Trichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl) ethane] 5,5,5-Tributyl phosphorotrithioate 0.(?-Diethyl 0-(2-isopropyl 4-methyl-6-pyrimidyl) phosphorothioate 2,6-DichIoro-4-nitroaniline 4,4'-Dichloro-ci-trichIoromethylbenzhydrol 3-(Dimethoxyphosphinyloxy)-/*/,N-dimethyl-c/5-crotonamide Not less than 85% of l,2,3,4,10,10-Hexachloro-6,7-epoxy-l,4,4a,5,6,7,8,8a-octahydro-l,4-en 65 4.40 ND 0.200 0.066 0.266 4,55 1.50 6.05 16 6 174 2.21 ND 0.022— 0048 0.070 0,93— 2.17 3.15 17 I 328 2.02 ND 0.(126 0.015— 0.041 — 1,29 0.74 2.03— 18 1 176 1.94 ND 0.059 0.027 0.086 3,04 1.39 4.43 19 4 69 4.08 ND 0.084 0.071 0.155 2.05 1.74 3.80 20 1 60 2.78 ND 0.127 0.045 0.172 4,57 1.62 6.19 NOTE: ND = not detected. — = lowest value. + — highest value. 152 Pesticides Monitoring Journal TABLE 10. Pesticide residues in human milk from district of Seliibal, Portugal — 1972 Number of Days Between Delivery and Fat On Whole-milk Basis, ppm On Fat Basis, ppm DDT Sample Deliveries Sampling Content, % DlELDRIN P,p'-DDE P.p'-DDT DDT P,p'-DDH P.p'-DDT Total 1 Unknown 56 0.41 ND 0.031 — 0.014— 0.045— 7.56 3.41 10.97 2 Unknown Unknown 1.98 ND 0.067 0.035 0.102 3.38 1.77 5.15 3 Unknown 8 2.27 ND 0.309 0.176 0.485 13.61 7.75 + 21.36 + 4 Unknown 238 5.27 ND 0.166 0.130 0,296 3.14— 2.47 5.61 5 Unknown 124 5.51 ND 0,220 0.085 0,305 3.99 1.54 5.53 6 Unknown 28 4.83 ND 0,403 0.183 + 0.586 8.34 3,79 12.13 7 5 11 1.38 ND 0,058 0.028 0.086 4.17 2.03 6.20 8 1 17 2.51 ND 0.087 0.047 0.134 3.48 1.87 5.35 9 2 186 2.02 ND 0,066 0.029 0.095 3.27 1.44 4.71— 11 1 210 2.57 ND 0.190 0.075 0.265 7.40 2.92 10.32 12 2 63 5.93 ND 0.229 0.076 0.305 3.86 1.28— 5.14 13 4 68 2.70 ND 0.112 0.061 0.173 4.13 2,26 6.39 14 3 164 1.17 0.006 0.038 0.024 0.062 3.28 2.05 5.33 15 1 40 2,97 ND 0.516 + 0.113 0.629 + 17.37 + 3.80 21.17 NOTE: ND = not detected. — =: lowest value. + — highest value. TABLE II. Pesticide residues in human milk from district of Viana do Castelo, Portugal — 7972 Number of Deliveries Days Between Delivery and Sampling Fat Content, % On Whole-milk Basis, ppm On Fat Basis, ppm Sample DlELDRIN p,p'-DDE P.p'-DDT Total DDT p,p'-DDE P.p'-DDT Total DDT 1 2 3 4 5 6 7 8 10 1 2 1 2 1 2 2 2 3 61 50 5 178 43 18 324 232 110 2.34 2.75 0.40 1.94 5.08 1.52 3.89 1.58 0.45 ND ND ND ND ND ND ND ND ND 0.072 0.076 0.020 0.123 0.146 + 0.042 0.083 0.019 0.007— 0.031 0.053 0.019 0.048 0.113 + 0.015 0.047 0.010 0.003— 0.103 0.129 0.039 0.171 0.259 + 0.057 0.130 0.029 0.010— 3.09 2.76 5.00 6.44 + 2.87 2.74 2.14 1.17— 1.56 1.32 1.93 4.75 + 2,47 2.22 0.99 1.21 0.63— 0.67 4.41 4.69 9.75 + 8.91 5.09 3.73 3.35 1.80— 2.23 NOTE: ND = not detected. — ^ lowest value. + = highest value. TABLE 12. Pesticide residues in human milk from district of Vila Real. Portugal — 1972 Number of Deliveries Days Between Delivery and Sampling Fat Content, % On Whole-milk Basis, ppm On Fat Basis, ppm Sample DlELDRIN P.p'-DDE p.p -DDT Total DDT P.p'-DDE P,p -DDT Total DDT 1 2 3 4 5 6 7 8 9 3 1 9 10 1 1 4 1 4 129 159 78 132 41 16 62 20 30 2.02 3.97 1.31 1.27 4.01 1.69 3.94 3.99 1.32 ND ND ND ND ND ND ND ND ND 0.043 0.100 0.025— 0.043 0.155 0.107 0.065 0.204 + 0.054 0.020 0.047 0.010— 0.024 0.058 + 0.044 0.033 0.048 0.055 0.063 0.147 0.035— 0.067 0.213 0.151 0.098 0.252 + 0.109 2.11 2,52 1.87 3.41 3.86 6.33 + 1.66 — 5.11 4.06 0.99 1.18 0.76— 1.89 1.44 2.60 0.84 1.20 4.17 + 3.10 3.70 2.63 5.30 5.30 8.93 + 2.50— 6.31 8.23 NOTE: ND = not detected. — = lowest value. + = highest value. Vol. 8, No. 3, December 1974 153 TABLE 13. Pesticide residues in human milk from district of Viseu, Portugal — 1972 Number of Days Between Delivery and Fat On Whole-milk Basis, ppm On Fat Basis, ppm Total Total Sample Deliveries Sampling Content, % Dieldrin P.P'-DDE P,P'-DDT DDT P.P'-DDE P.P'-DDT DDT 1 3 65 1.67 ND 0.066 0.042 0.108 3.92 2.51 6.43 2 4 88 1.85 ND 0.021 0.018 0.039 1.14 0.97 2.11 3 4 208 1.40 ND 0.015 0.011 0.026 1.04 0.79 1.83 5 1 18 1.91 ND 0.036 0.012 0,048 1.86 0.63 2.49 6 1 17 3.42 0.009 0.057 0.057 0.H4 1.67 1.67 3.34 7 4 193 3.71 0.005 0.008 0.009 0.017 0.22— 0.24 — 0.46 — 8 1 90 1.05 ND 0.027 0.013 0.040 2.57 1.24 3.81 9 2 128 3.63 0.017 0.032 0.016 0.048 0.87 0.44 1.31 11 1 164 2.19 ND 0.044 0.022 0.066 1.99 1.00 2.99 12 1 218 1.84 0.015 0.021 0.016 0.037 1.13 0.85 1.98 13 6 164 0.80 0.005 0.006— 0.004— 0.010 0.62 0.44 1.06 14 2 83 1.81 ND 0.020 0.020 0.040 1.10 1.11 2.21 15 2 18 2.75 ND 0.109 -L 0.158 + 0.267 + 3.97 5.73 + 9.70 + 16 2 93 0.96 ND 0.041 0.014 0.055 4.22 + 1.46 5.68 17 1 42 3.19 ND 0.047 0.025 0.072 1.47 0.78 2.25 18 3 69 2.54 0.021 ND ND 0.01 — ND ND 19 1 20 1.49 ND 0.029 0.028 0.057 1.92 1.88 3.80 20 5 22 2.12 ND 0.018 0.011 0.029 0.83 0.52 1.35 NOTE: ND = not detected. — = lowest value. + = highest va ue. Tables 14 and 15 show that lowest mean levels of DDT were found in the rural districts Braganca and Viseu. The highest mean level was found in the district of Lisbon. Various other authors have reported lower contamination levels in rural areas than in urban areas (16.18.21). Kroger (/2) observed that mothers nursing firstborn children showed higher levels of DDT than did other mothers. This may account for the difference observed between mean levels in nursing mothers in the district of Lisbon and those in Braganga, but could hardly justify the difference in relation to Viseu. However, it is not only the number of deliveries that influences an individual's contamination levels, but also the number of days separating sampling from delivery, and the indi- vidual's dietary habits. Authors have not drawn conclu- sions regarding the influence of each of these factors. Dieldrin was detected in 15 of 222 samples examined; levels ranged from 0.006 to 0,031 ppm. The district of Viseu showed the highest percentage of samples con- taining this insecticide; it was found in 33 percent of the samples collected in that area. In Braganga. 20 per- cent of the samples had dieldrin residues. Similar diel- drin levels have been reported by various authors (.6,7.10,13,16,19-22). Most significant is the Japanese study in which residues of this insecticide were found in 74.6 percent of 398 samples of human milk (16). In Australia. Newton observed that in 67 samples exam- ined, about 43 percent contained dieldrin residues (18). Apart from DDT and dieldrin, BHC isomers are the pesticides which have been most widely reported in human milk. In the present case, 7 BHC was detected in some samples but, because of its low level, was not measured. This was the only isomer reported by Westoo et al. in human milk in Sweden (10) but the (3 isomer has been detected at very high levels by various authors (13.16.19-21). TABLE 14. Total DDT residues in human whole milk, Portugal~1972 Residues, mo/kg Standard Deviation Number OF' DELIVERIES' District Minimum Maximum Mean Aveiro 0.047 0.743 0.283 -1-0.265 2.1 ^ragan^a 0.010 0.149 0.063 -t-0.034 4.4 Evora 0.031 0.837 0.269 -<-0.241 1.7 Faro 0.038 0.272 0.146 -^0.094 1.6 Guarda 0.072 0.366 0.165 -*-0.106 2.6 Lisbon 0.083 0.780 0.326 -1-0.175 1.6 Porto 0.041 0.353 0.136 -^0.088 Unknowns Portalegre 0.057 0.606 0.256 -1-0.181 2.1 Setubal 0.032 0.629 0.252 -1-0.197 Unknownr Vila Real 0.035 0.252 0.126 -*-0.072 3.8 Viana do Cas telo 0.010 0.259 0.103 -<-0.079 1.8 Viseu <0.010 0.343 0.064 ±0.075 2.4 TABLE 15. Converted mean of total DDT residues in human whole milk submitted to the^i/x transformation, Portugal— 1972 County or Converted Significance District Mean, mg/kg Level, % ' Odivelas - 0.4778 Amadora - 0.3831 Loures - 0.2570 Cane?as - 0.2396 Damaia - 0.2163 Portalegre 0.2122 Aveiro 0.2067 Evora 0.2051 Setubal 0.1962 Guarda 0.1453 5 Faro 0.1243 NS Porto 0.1178 5 Vila Real 0.1121 NS Viana do Castelo 0.0798 NS Viseu Braganca 0.0415 0.0564 NS NOTE: NS = not significant. ' Significance levels determined by the Dunnet test (Dunnet, 1964). = Towns in the district ot Lisbon. In the present study of human milk in Portugal, sam- ples also contained traces of HCB which were not measured. HCB was detected in human milk in Aus- tralia (18) and in France (13,20). 154 Pesticides Monitoring Journal TABLE 16. Professions of hictaling women whose milk was analyzed, Portugal — 1972 Number of Mothers District Questioned HOMEMAKERS Civil Servants Factory Workers Farmworkers Other > Aveiro 10 9 1 Bragan?a 19 19 Evora 10 8 1 Faro 5 4 1 Guarda 10 9 1 Lisbon 29 25 1 2 Porto 22 20 Portalegre 10 9 Setiibal 15 14 1 Vila Real 10 8 1 Viano do Castelo 10 10 Viseu 20 19 ' Dressmakers, office assistants, and shop assistants. Some authors have also detected lindane (7.10,22-23), heptachlor epoxide (6,7,13,20,22.23). endrin (16), al- drin (16). and PCB's (10.19), but none of these prod- ucts was detected in human milk in Portugal. PCB's were not even sought during this work. Comparing mean residues of total DDT detected in each of the districts of Portugal with values published in other countries and tabulated in a paper presented by Ritcey et al. (22), values found in Portugal are all higher than those in the Netherlands (0.048 ppm) and are all lower than those published in works on Romania (0.530 ppm) and Poland (0.40 ppm). It may be concluded that mean residues of total DDT in Portu- gal were generally similar to those of other European and North American countries. The sale of DDT, BHC. and heptachlor was for- bidden in Portugal after January 1, 1974. and the use of aldrin, dieldrin, and endrin has been greatly restricted. It is hoped that the problem of unintentional contami- nation by organochlorine pesticides in our country will begin to right itself, however slowly. Acknowledgment Authors wish to thank personnel of the Maternity Institute in Lisbon (Instituto Maternal. Direccao-Geral da Assistencia Social, Lisbon), especially Dr. Maria Fernanda Navarro, for milk samples collected in dif- ferent districts of Portugal. LITERATURE CITED (1) Lang, E. P., F. M. Kunze, and C. S. Prickell. 1951. Occurrence of DDT in human fat and milk. Arch. Industr. Hyg. 3(3 ): 245-246. (2} De Faubert Maunder, M. ]., H. Egan. E. W. Godly, E. W. Hammond, J. Rohurn, and J. Thomson. 1964. Cleanup of animal fats and dairy products for the analysis of chlorinated pesticide residues. Analyst 89; 168-174. (3) Jeffers, J. N. R. I960. Experimental design and analysis in forest research. Almqvist and Wiksell, Stockholm. 14) Dunnet, C. W. 1964. New tables for multiple compari- son with a control. Biometrics 20:482-491. (5) Qiiimhy, G. E., J. F. Armstrong, and W. F. Durham. 1965. DDT in human milk. Nature 207:726-728. (6) Curley, A., and R. Kimhrough. 1969. Chlorinated hy- drocarbon insecticides in plasma and milk of preg- nant and lactating women. Arch. Environ. Health 18:156-164. (7) Hcyndrickx, A., and R. Maes. 1969. The excretion of chlorinated hydrocarbon insecticides in human mother milk. J. Pharm. Belg. 24:459-463. (8) Gracheva, G. V. 1969. The possibility of DDT accu- mulation in the organism of persons not having occu- pational contact with it. Faktory Vneshn. Sredy i ikh Znachen. dlia Zdorov'ia Naselen 1:125-129. Cited from Health Aspects Peslic. Abstr. Bull. 1970, 3:1304. (9) Komorova, L. I. 1970. DDT excretion with the breast milk and its effect on the body of the mother and child. Pediatr. Akusherstvo Hinekol. 35(1): 19-20. Cited from Health Aspects Pestic. Abstr. Bull. 1971, 4:474. (10) Westoo, G., K. Noren, and M. Anderson. 1970. The levels of organochlorine pesticides and polychlorinated biphenyls in margarine, vegetable oils, and some foods of animal origin on the Swedish market in 1967-1969. Klorpesticid-och polyklorbi fenylhalter i margarin, vegetabila matoljor och vissa animala livsmedel i svensk handel aren 1967-1969. Var foda 22(2-3): 9-3L (11) Tiiinstra, L. G. M. Th. 1971. Organochlorine insecti- cide residues in human milk in the Leiden region. Ned. Melk-Zuiveltijdschr. 25(l):24-32. Cited from Health Aspects Pestic. Abstr. Bull. 1971, 4:2118. (12) Kontek, M., S. Kubacki, S. Paradowski, and B. Wierz- chowiecka. 1971. Study of the level of organochlorine pesticides in human milk. Badanie zawartosci pestycy- dow chloroorganicznych w mieku kobiecyni. Pediat. Pol. 46(2); 183-188. Cited from Health Aspects Pestic. Abstr. Bull. 1971, 4:1619. (13) Goursaud, J., F. M. Litquet, and J. Casalis. 1971. Pesti- cide residue contamination of human milk in the northern provinces of France and Pas-de-Calais. Sur la pollution des laits de femme par les re'sidues de pesticides dans les departements du Nord et du Pas- de-Calais. Lait 51:559-567. il4) Dymeni, P. G., L. M. Hebertson, E. D. Gomes, J. S. Wiseman, and R. W. Hornabrook. 1971. Absence of Vol. 8, No. 3, December 1974 155 polychloriiiated biphenyls in human milk and serum from Texas and human milk from New Guinea. Bull. Environ. Contam. Toxicol. 6:532-534. (15) Sugaya, T., el al. 1971. Organochlorine pesticide resi- dues in human milk. Nippon Noson Igakkai Zasshi 19(4):379-380. Cited from Health Aspects Pestic. Abstr. Bull. 1972, 5:1644. (16) Anonymous. 1972. BHC and DDT residues in human milk on the decline in Japan. Noyaku Bijinesu (Pestic. Business) 56:458. Cited from Health Aspects Pestic. Abstr. Bull. 1972, 5:1660. (17) Jiiszkiewicz, T., J. Stec, T. Radomanski, and B. Tre- bicka-Kwiatkowska. 1972. Residues of organochlorine insecticides in the colostrum and milk of women after delivery Pol. Tyg. Lek. 27(17) :616-619. Cited from Health Aspects Pestic. Abstr. Bull. 1972, 5:1875. (18) Ncwion, K. G., and A'. C. Greene. 1972. Organochlo- rine pesticide residue levels in human milk — Victoria, Australia— 1970. Pestic. Monit. J. 6(l):4-8. (/9) Hidaka, K., T. Ohe, and K. Fujiwara. 1972. PCB and organochlorine pesticides in mother's milk. Igaku No Ayumi (Progr. Med.) 82(8 ) :519-520. Cited from Health Aspects Pestic. Abstr. Bull. 1972, 5:2306. (20) Lnqiiel, F. A/., J. Goursaud, and B. Gaudier. 1972. Study of the pollution of human milk by residual pesti- cides. Etude de la pollution des laits humains par les residues de pesticides. Palhol. Biol. 20:137. (21) Kuroda, H., T. Yano, K. Kagana, and M. Mit.'imune. 1972. On the residual organochlorine pesticides in mother's milk. Shikoku Koshu Eiseigakkai Zasshi (J. Shikoku Pub. Health Soc.) 17:79-80. Cited from Health Aspects Pestic. Abstr. Bull. 1972, 5:2331. (22) Rilcey. W. R.. G. Savary, and K. A. McCully. 1972. Organochlorine insecticide residues in human milk, evaporated milk and some milk substitutes in Canada. Can. J. Pub. Health 63:125-133. (23) Kroger, M. 1972. Insecticide residues in human milk. J. Pediat. 80:401-405. 156 Pesticides Monitoring Journal PESTICIDES IN WATER A Study of the Distribution of Poly chlorinated Biphenyls in the Aquatic Environment^ Hans J. Crump-Wiesner,2 Herman R. Feltz,3 and Marvin L. Yates ^ ABSTRACT Data gathered from monitoring activities and project studies indicate the uhiquiious occurrence and distribution of polychlorinated biphenyls (PCB's) in the aquatic environ- ment. By 1972 residues had been detected in samples from 19 States representing nearly every region of the country. These findings permit a preliminary assessment of PCB con- tamination across the Nation: concentrations ranged from 0.1 to 4.0 ng/liter in unfiltered water samples and from 5.0 to 3,200 tig/kg in bottom sediments. PCB residues were also found in fish and aquatic plants. Samples were prepared by the same techniques used for general chlorinated insecti- cide detection, with special attention to cleanup and separa- tion of PCB's from other compounds. Basic identification and quantification were made by dual-column electron- capture/ gas-liquid chromatography and confirmed by gas- liquid chromatography /mass spectrometry whenever pos- sible. In sediment samples from a south Florida drainage ditch, polychlorinated naphthalenes (PCN's) were observed. This is possibly the first evidence of PCN's in an environ- mental sample and illustrates the importance of developing analytical capability for the surveillance of other organo- chlorine compounds that may behave like chlorinated hydro- carbon pesticides. The sampling program is broadening geographically and gradually increasing to more adequately define the distribu- tion of PCB residues in major drainage basins of the United States. > Presented to the Division of Pesticide Chemistry, 164th National Meeting of the American Chemical Society, New York, N.Y., August 29, 1972. Subsequently published in the Journal of Research of the U.S. Geological Survey, Vol. 1, No. 5. Reprinted here with permission because data are considered significant to readers of the Pesticides Monitoring Journal. ' Office of Water Programs Operations, U.S. Environmental Protection Agency, Washington. D.C. 20460. ^ Water Resources Division, Northeastern Region, Geological Survey, U.S. Department of Interior, Reston. Va. * Water Resources Division, Western Region, Geological Survey, U.S. Department of Interior, Menlo Park, Calif. Introduction Within the past few years, polychlorinated biphenyls (PCB's) have been identified as a major environmental contaminant. Their detection has caused widespread concern and has generated intense interest in data relat- ing to the presence and effects of these compounds, par- ticularly in the aquatic environment. First produced about 40 years ago. PCB compounds have become in- creasingly useful in such industrial applications as com- ponents in transformers and capacitors, heat exchangers, paints, inks, dyes, and dust control agents. Although much attention has been focused on estimat- ing levels and potential hazards of PCB's in aquatic organisms, few data are available on the occurrence of PCB's in water and bottom sediments. The Geological .Survey. U.S. Department of Interior, through its water- qtiality-monitoring activities and water-resources-assess- ment projects, has been alert to the PCB problem since it was first reported by Widmark in 1967 (I). Although the Geological Survey has no nationwide PCB assess- ment program, sufficient data have accumulated from pesticide residue programs to permit a preliminary as- sessment of PCB contamination of the Nation's hydro- logic environment. This paper presents data gathered from these activities, showing the widespread occur- rence of PCB's in significant concentiations in unfiltered surface water and ground water, bottom sediments, Hora. and fauna. Analytical Techniques PCB residues were analyzed by the multiple-pesticide- residue methods for water, suspended sediment, and bot- tom material described by Goerlitz and Brown (2). Analytical procedures are appropriate for chlorinated pesticides as well as the general class of organochlorine Vol. 8, No. 3, December 1974 157 compounds. Special attention was given to cleanup and separation ot PCB's from coextractives. EXTRACTION OF WATER, SEDIMENT, AND BIOTA One-liter unlilteied water samples were collected in pre- cleaned glass bottles and extracted three times with hexane. The hexane portions were combined, dried with anhydrous Na.,SO|, and concentrated to 1 ml before cleanup and analysis by electron-capture/ gas-liquid chromatography (EC/GLC). Fifty-gram sediment samples (dry-weight basis) were extracted with an acetone-hexane solvent. The sediment was dispersed first in acetone, and hexane, was added to recover the acetone and the desorbed material. The extract was washed with distilled water, dried over Na^.SO^, and concentrated to 5 ml for cleanup before EC/GLC analysis. The two extraction procedures used for biota are de- scribed in an analytical manual issued by the Food and Drug Administration, U.S. Department of Health, Edu- cation, and Welfare (3). Fish samples were extracted with petroleum ether in a blender; aquatic plants were extracted with acetonitrile. The chlorinated hydrocarbon fraction was partitioned between petroleum ether and acetonitrile, dried, and concentrated to a final volume of 5 ml. .SEPARATION Bottom sediment, fish, and plant extracts were cleaned following the Law and Goerlitz technique (4) requiring less time and smaller volumes of solvents for elution than do other widely used methods. Liquid/solid col- umn chromatography was employed using two different types of semimicro columns in sequence (Fig. 1). Hex- ane extracts were fiist passed through an alumina col- umn, and one fraction was further chromatographed on silica gel to separate PCB's from chlorinated insecti- cides. Successive chromatography on alumina and silica gel results in a simultaneous cleanup and separation of PCB's from the common insecticides, except aldrin, and a slight overlap of p.p'-DDE. To achieve a reduction in background interference, mercury was added to remove sulfur from bottom sediment extracts before they were applied to the silica column. In order to insure reproducible chromatographic con- ditions, the activity of the adsorbents was carefully con- trolled. Water extracts were cleaned on a deactivated alumina microcolumn (5). When PCB's were detected in the cleaned water extracts, they were also separated on a semimicro silica gel column. IDENTIFICATION Basic identification was made by dual-column EC/GLC (DC-200 and QF-l/OV-17) and confirmed by gas- liquid chromatography/ mass spectrometry (GLC/MS) when sample size and concentrations were sufficient. The amount of PCB's was determined by matching the unknown peaks on the chromatogram to the nearest commercial formulation and measuring the areas of four corresponding peaks. Retention time and peak area measurements were made with a digital electronic inte- grator. The lower detection limit for PCB residues was 0.1 ug/liter in water and 5.0 |ig/kg in bottom sediment. Reported levels are subject to considerable error because of the complexity of multiple peaks, some peak altera- tion, and the occasional presence of mixtures of PCB's in environmental samples. At best, reported values are estimates that may be as much as 50 percent in error. H exane extract Alumma 1 " ' 1 0 — 20 ml fraction 20 — 35 ml fraction 35—50 ml fraction (hexane) (hexane) (benzene) PCBs Di«(drin Ethion PCNs En dnn Malath.on Aldnn H«ptachlor «poxide Methyl parsthion Chlordsne Parathion DOD Methyl tnthion DDE DDT HeplKhlor Lindane Toxaphene 1 1 Silica gel 1 1 \ 0—25 ml fraction 25—45 ml fraction (hexane) (benzene) PCBs Chlordane PCNs DDD Aldrin DDE DDT Heptachlof Lindane Toxaphene FIGURE 1. Scheme for separating polychlorinated bi- phenyls (PCB's) and polychlorinated naphthalenes (PCN's) from pesticides DATA DESCRIPTION Occurrences of pesticide residues in the aquatic environ- ment have been documented over a period of years through monitoring programs of several Federal agen- cies. As early as 1957: studies of chlorinated hydro- carbon pesticides in major river basins were made by the Federal Water Pollution Control Administration, now a part of the U.S. Environmental Protection Agency, by use of the carbon-adsorption technique (6). In 1964 interagency cooperation in pe*;ticide monitoring programs culminated in a proposal to begin a national monitoring program. The original program for water was described in 1967 in the first issue of the Pesti- cides Monitoring Journal (7). The purpose of this pro- gram, which was revised in 1971 (8). is to provide con- tinuing information on levels of pesticide residues in the water -resources of the Nation and to identify pos- sible problem areas. Because 'PCB's are analyzed by the 158 Pesticides Monitoring Journal multiple-pesticide residue techniques, routine reporting ol these compounds has been incorporated into current pesticide programs. At present, samples from 20 of the 161 proposed network sites are collected and analyzed by the Geological Survey. All samples are collected from sites west of the Mississippi River and provide continuity with a network established to evaluate the quality of water used for irrigation (9,10). Budgetary restrictions have prevented further implementation of the network. After development of the technique to separate PCB's from pesticide residues, examination for the presence of PCB's in water and suspended and bottom sediment samples collected for the National Monitoring Program began in January 1971. Funding was requested to in- crease the number of network stations to 50 in 1973, and to 100 in 1974. allowing a better assessment of PCB's and pesticides in major drainage basins through- out the United States. Analysis of 194 water samples and 33 bottom sediment samples revealed no positive identifications of PCB's; however, these data are not truly representative of the entire Nation because of the limited number of sites sampled. In 1958. the Geological Survey began operation of a bench-mark network to provide basic hydrologic data on selected stream basins throughout the United States that arc expected to remain in their present natural condition or are not expected to be significantly altered by humans. Locations of the 57 bench marks estab- lished in 37 States are shown in Figure 2. To insure mininiLim interference by humans, many of the hydrologic bench marks are in national parks, wil- derness areas. State parks, national forests, and areas set aside for scientific study. A detailed description of the network basins, including drainage, climate, topog- raphy, geology, vegetation, hydrology, water quality, and personmade influences, appears in a report by Cobb and Biesecker {11). Data gathered from 46 of these sites are presented in Table 1 . Despite the careful screening for pristine location of bench-mark sites, two bottom sediment samples analyzed in the 1972 water year contained PCB residues. A value of 5.2 |.ig/kg was measured in the sample from South Fork Rocky Creek near Briggs, Tex., and 8.8 |ig/kg was found in the sample from Upper Twin Creek at McGaw. Ohio. The majority of the Geological Survey water resources studies are conducted in cooperation with State water resources and pollution control agencies, or in response to requests from other Federal agencies. PCB data from these programs in 35 States are presented in Tables 2-4. HAWAII ^^^~\ FIGURE 2. Map showing hydrologic bench-mark stations. (Numbers refer to list in Cobb and Biesecker: refer to Literature Cited, reference 1.) Vol. 8, No. 3, December 1974 159 TABLE 1. Summary of PCB residue data, natiomd hydro- logic bench-mark network, January 1971-June 1972 TABLE 4. Summary of PCB residue data from selected sampling sites in Florida, January 1971 -J line 1972 Type of Sample Unit No. Samples Occur- rences CONCEN- T RATION Water Bottom sediment /lig/kg 54 51 0 2 5.2, 8.8 TABLE 2. Summary of PCB residue data for surface and ground water, January 1971-June 1972 State Alaska Arizona Arkansas California Colorado Connecticut Hawaii Iowa Kansas Kentucky Louisiana Maine Maryland Massachusetts Michigan Minnesota Mississippi Missouri Montana Nebraska New Jersey New Mexico New York North Dakota Oklahoma Oregon Pennsylvania Puerto Rico South Dakota Texas Virginia Washington West Virginia Wisconsin Wyoming No. Samples 3 8 32 161 32 13 5 24 10 7 9 2 6 5 2 3 8 21 47 44 11 35 325 40 19 13 2 7 18 660 4 25 4 3 18 Occur- rences 0 0 0 2 1 6 0 0 0 0 0 0 1 1 0 2 0 0 0 0 3 0 52 (1 0 0 1 1 0 12 1 0 0 0 0 Concen- tration, „g/LITER ND ND ND 0.1,0.1 0.3 0.1-0.2 ND ND ND ND ND ND 0.1 0.2 ND 0.1.0.3 ND ND ND ND 0.1 ND 0.1-4.0 ND ND ND 0.2 0.1 ND 0.1-3.0 0.1 ND ND ND ND Median Concentration. „g/LITER ND ND ND ND ND 0.1 ND ND ND ND ND ND ND ND ND ND ND ND ND ND 0.1 ND 0.3 ND ND ND ND ND ND 0.4 ND ND ND ND ND NOTE: ND = not detected. TABLE 3. Summary of PCB residue data for bottom sedi- ments, January 1971-June 1972 Concen- MEDIAN State No. Occur- tration. Concentration, Samples rences ug/kg «g/kg Alaska 3 0 ND ND Arkansas 3 4 20-2,400 60 California 13 3 20-190 85 Connecticut 1 1 40 ND Hawaii 4 0 ND ND Georgia 12 10 10-1.300 300 Maryland 11 5 10-1,200 30 Mississippi 8 2 50:170 ND New Jersey 12 10 8-250 20 Oregon 4 T !5;140 ND Pennsylvania 16 11 10-50 20 South Carolina 11 8 30-200 50 Texas 293 23 7.9-290 80 Virginia 10 8 5-80 40 Washington 10 0 ND ND West Virginia 2 1 10 ND Concen- MEDIAN Type No. Occur- tration, Concen- Sample Samples rences Range i tration ' Water 231 12 0.1-2.1 0.2 Bottom sediment 118 50 5-3,200 30 Flora 16 5 10-50 20 Fauna 43 36 6-1,000 40 NOTE: ND = not detected. ' Unit of measure: ;xg/liter for water samples, ^ug/kg for other samples. Water samples alone, because of the low water solubility of PCB's, are not a good indicator of the widespread occurrence of the compounds, and show an incidence of slightly over 5 percent. Unfiltered water samples from 12 of the 35 States had PCB concentrations rang- ing from 0.1 to 4.0 ug/liter. However, a significant number of suspected traces of PCB's were not reported because of several analytical limitations. Generally, re- sampling of areas where PCB's were first detected re- vealed that the compounds were still present several months later. Bottom sediments were collected concurrently with many of the water samples reported in Table 2. These samples were taken from lakes and streams that drain a variety of land-use areas generally located away from industrial centers. Data in Table 3 show that bottom sediment samples may be used as an indicator of PCB contamination in the Nation's hydrologic environment. Significantly, of samples collected at random from 16 States. 13 contained PCB's in the range of 5.0 to 2.400 l^ig/kg. Across the Nation, one of every five bottom sediment samples contained PCB's. Data available from Florida (Table 4) merit special attention because they reveal the distribution of resi- dues in several environmental components. Only 12 of 231 unfiltered water samples contained PCB's ranging from 0.1 to 2.1 ug/liter. but over 40 percent of the associated bottom sediments analyzed during the same period were contaminated with PCB's ranging from 5.0 to 3.200 |ig/kg. Median PCB concentrations of 20 iig/kg and 40 iig/kg found in aquatic plants and fish, respectively, follow the scheme of the biological accumulation of the DDT family in southern Florida (72). Results and Discussion Preliminary data presented in this report indicate that significant concentrations of PCB's are widespread in the water resources of the Nation. However, there are some shortcomings of data compiled from pesticide resi- due programs. First is the problem of nonrepresentative sampling within States and some repetition of sampling in a given basin. Second, the lower limit of detection for PCB residues on the basis of a one-liter water sample is inadequate for critical evaluation. Trace amounts of less than 0.1 pg/ liter were detected in a 160 Pesticides Monitoring Journal significant number of samples but were excluded from the tabulation because they could not be confirmed. Third, because of the low solubility of PCB's in water, especially the higher chlorinated ones, the bulk of PCB residues in streams are associated with suspended sedi- ment and bottom material. Therefore. PCB concentra- tions may be expected to vary directly with the sus- pended sediment concentration in the cross section of a stream. Most surface water samples were collected by depth integration at the center of flow, which usually does not provide a representative sample of suspended sediment. Future investigations should be made with a depth-integrating sampler using the equal-transit-rate procedure (12). In spite of these shortcomings, evi- dence for the ubiquity of PCB's in the hydrologic en- vironment is clearly established. As others have pointed out (13), polychlorinated naph- thalenes (PCN's) are compounds that have uses similar to PCB's and may possibly be present in environmental samples. They can be separated from chlorinated hydro- carbon insecticides and are eluted in the same fraction as PCB's by alumina/silica gel column chromatography. Sediment samples collected from a south Florida drain- age ditch contained mixtures of PCN's ranging from 1,250 to 5,000 jig/kg, whereas water samples overlying the sediments averaged 5.7 j^tg/liter. Identification was confirmed by both microcoulometry and GLC/MS. This is possibly the first evidence of PCN's in an en- vironmental sample and illustrates the importance of developing analytical capability for the surveillance of other organochlorine compoimds that may behave like chlorinated hydrocarbon pesticides. It is probable that PCB's enter the aquatic environ- ment through low-temperature incineration of solid wastes, industrial waste disposal into waterways, and sewage outfalls. The highest levels are usually associated with industrial areas and nearby aquatic food chains. In contrast, authors have noticed significant concentra- tions in the bottom sediments of drainage ditches, multi- purpose canals, and suburban real estate lakes remote from major industrial and metropolitan areas. The pres- ence of PCB's in real estate lakes has been attributed to a variety of construction materials used in the hous- ing industry. PCB's in surface and ground water used for public water supplies were detected through coop- erative programs. The highest concentration was 4.0 |xg/liter in an untreated source for a city in the State of New York. It is clear that the presence of PCB's and other organo- chlorine compounds in the aquatic environment merits continuing observation because of the limited evaluation that can be made from the meager data available. It is important to measure baseline levels of PCB's in streams and lakes in order to determine trends. Long- term monitoring on a systematic basis will provide the data necessary to assess the presence of PCB residues and concurrently reveal problem areas. LITHRATURE CITED (/) Wictinark, G. 1967. Pesticide residues, possible inter- ference by chlorinated biphenyls. In Proceedings of the TUPAC Commission on the Development, Im- provement, and Standardization of Methods of Pesti- cide Residue Analysis, J. Ass. Offic. Anal. Chem. 50(5): 1069. (2) Goertitz, D. F., and Eugene Brown. 1972. Methods for analysis of organic substances in water. U.S. Geol. Survey Techniques Water Resources Inv. TWI 5-A3. 40 pp. (Book 5, Laboratory Analysis.) (3) U.S. Department of Health, Education, and Welfare — Food and Drug Administration. 1971. Pesticide an- alytical manual. Vol. 1. (4) Goertitz, D. F., and L. M. Law. 1974. Determination of chlorinated insecticides in suspended sediment and bottom material. I. Ass. Offic. Anal Chem. 57(1): 176- 181. (5) Law, L. M., and D. F. Goerlitz. 1970. Microcolumn chromatographic cleanup for the analysis of pesticides in water. J. Ass. Offic. Anal. Chem. 53:1276-1286. (6) Breidenhach, A. W., et al. 1964. The identification and measurement of chlorinated hydrocarbon pesticides in surface waters. U.S. Pub. Health Serv. Publ. 1241. 70 pp. (7) Green, R. S., and S. K. Love. 1967. Network to moni- tor hydrologic environment covers major drainage rivers. Pestic. Monit. J. 1(1): 13-16. (8) Feltz, H. R., W. T. Savers, and H. P. Nicholson. 1971. National monitoring program for the assessment of pesticide residues in water. Pestic. Monit. J. 5(1): 54-62. (9) Brown, Eugene, and Y. A. Nishioka. 1967. Pesticides in selected western streams — a contribution to the National Program. Pestic. Monit. J. 1(2): 28-46. (10) Manigold, D. B., and J. A. Schulze. 1969. Pesticides in selected western streams — a progress report. Pestic. Monit. 1. 3(2):124-135. (7/) Cobb, E. D., and J. E. Biesecker. 1971. The national hvdrologic bench-mark network. U.S. Geol. Surv. C'irc. 460-D. 38 pp. (12) Feltz, H. R., and J. A. Culbertson. 1972. Sampling procedures and problems in the determination of pesti- cide residues in the hydrologic environment. Pestic. Monit. J. 6(3): 171-178. (13) Goerlitz. D. F., and L. M. Law. 1972. Chlorinated naphthalenes in pesticide analysis. Bull. Environ. Con- tarn. Toxicol. 7:243-251. Vol. 8, No. 3, December 1974 161 RESIDUES IN FISH, WILDLIFE, AND ESTUARIES Chlorinated Hydrocarbon Pesticide Residues in Oysters (Crassostrea commercialis) in Moreton Bay, Queensland, Australia, 1970-72 ^ Donald Edward Clegg ABSTRACT A 2-year survey was carried out to monitor levels of DDT, ODD, DDE, and dieldrin in oysters (Crassostrea commercialis) in Moreton Bay, Queensland, Australia. Samples were taken at quarterly intervals from eight sta- tions located at or near the mouths of streams entering lite bay. Highest levels of 0.94 ppm DDT, 0.51 ppm DDD, 0.20 ppm DDE, and 0.34 ppm dieldrin were found in July 1970 at the sampling station on the Brisbane River 16 km down- stream from the center of Brisbane, a city of 700,000. Maxi- mum values at other stations were substantially lower. Residue levels varied considerably throughout the 2-year period. Authors attribiUe this at least in part to seasonal rainfall patterns in the catchment areas. Introduction A program to monitor chlorinated hydrocarbon pesti- cide residues in oysters (Crassostrea commercialis) in Moreton Bay, Queensland, Australia, was begun in April 1970 and continued approximately at quarterly intervals until March 1972. The oyster was chosen as an indicator organism partly because of its ubiquity and immobility but mainly because Butler et al. have shown that it has a marked ability to concentrate certain chlorinated hydrocarbons from the surrounding water and, when subsequently placed in clean water, is able to purge itself rapidly of these compounds (/). These characteristics are useful not only in helping to deter- mine geographical distribution of pesticides in this region but also in facilitating observation of seasonal variations of pesticide levels, particularly as a result of changes in the flow volume of fresh water in adjacent streams. Moreton Bay is formed by two large sandy islands, Moreton and Stradbroke. The bay is fed by several rivers, the chief being the Brisbane River, and a number of smaller streams. The southern end is estuarine and the north coast faces relatively open water (Fig. 1). ' Animal Research Institute, Department of Primary Industries. Queens- land, Australia. FIGURE 1. Map of Moreton Bay area, Queensland, Aus- tralia, surveved for pesticide contamination in oysters, 1970-72 162 Pesticides Monitoring Journal The cil\' unci suburbs of Brisbane occupv much of the central region ol Nforeton Bay. exlcnding about 24 km inland. Industry is concenlrated along the hanks of the Brisbane Ri\'er. especiallv toward the nioLith. .Smaller oulUing towns, such as Caboolture. Clevelantl, and Beenleigh. are centers lor Iruit. vegetable, sugar cane, and dairy larmmg; they also serve as dormitory towns lor city commuters. The bay waters yield commercial quantities of fish, prawns, crabs, and oysters. Although there was little evidence that viability of these marine- based industries has been alTcctcd by proximity of an urban-industrial center and surrounding intensive agri- cultural areas, it was felt that a survey providing infor- mation dealing with extent and distribution of persistent pesticides in the bay would provide data against which future investigations might be compared. It would also allow comparisons to be made between pesticide residue levels in this area and those in other regions. Sampling Sampling points were located as near as possible to mouths of streams flowing into the bay in the hope of determining the relationship between residue levels in the oysters and the land use of the catchment area (Fig. n. All oysters were collected from rocks or man- grove roots in the intertidal zone. Table I describes in- dividual sites and land use and population of the catch- ment area. Analytical Procedures Oysters were collected in their shells and kept on ice during transportation to the laboratory where they were stored at — 4°C pending analysis. Whole oyster-meat samples of approximately 10 g were submitted to the cleanup procedure for nonfatty foods described in the Pesticide Analytical Manual Vol. 1. Section 212.13a (1971) published by the Food and Drug Administration, U.S. Department of Health. Education, and Welfare. Appropriate adjustments were made for sample size. An aliquot of homogenized meat was extracted with acetonitrile and partitioned between hexane and the acetonitrile extract, which was diluted with water. The hexane phase was back-washed with water, dried, and cleaned in a florisil column packed with activated magnesium silicate. A Varian 1400 gas chromatograph equipped with a tritium-foil electron-capture detector was used to identi- fy and quantify pesticide residues on the sample ex- tracts. Operating parameters were as follow: Column: DC-200 5 percent on Aeropalc 30 Temperalures: detector, 2()n°C; injector, 240°C; cotiimn. 185°C Carrier gas: nitrogen 40 ml min Recoveries of DDT. DDD, DDE. and dieldrin from samples ' fortified with 1 iig of each compound were 98. 93, 96, and 83 percent, respectively. Blank runs on solvents and reagents allowed a lower limit of reliable estimation to be set at 0.005 ppm. All data reported are corrected lor recovery. For confiiniation of peak identity, the extract was concentrated to a low volume and a thin-layer chro- matogram was rtin and compared with standards treated in a similar manner. At the levels under consideration, visualization of the spots was not possible and com- ponents were identified by scraping zones from the plate, extracting with hexane/ether. and injecting the extract into the gas chromatograph. T,-\BLE 1. Aimitiliiin sunipliiif; sites, land iiw. and poptdn- liim of catiltmciit areas from which oysters nere collected for monitoring, 1970-72 Station 1 Site: adjacent to small boat harbor. Samples from rock wall. Indiistr.v: dairy farming. Population: low-density riirat, with small holiday settlement. Station 2 Site: rocky headland adjacent to estuarine area of bay. Samples from rock. Ini.luslr\: ^iigar-cane farming, dairy farming, sugar mill, Uisiijlery, Ptipiilation: predommantly rural; one town. Beenleigh. Station 3 Site: adjacent to small boat harbor. Samples from rocks. Industry: fruit and vegetable farming. Population: intensive rural; some outer-city suburbs. Station 4 Site: rocky headland near small boat harbor. Samples from rocks. Industry: principally suburban residential. Population: medium- to high-density urban. Station 5 Site; rock wall on river's edge near oil refinery. Samples from rocks. Industry: dairying and intensive agriculture in upper reaches of river. Industrial, commercial and high-density urban in lower reaches. Population: high density; major city, Brisbane. Station 6 Site: Industry: Population: rocky headland adjacent to small boat harbor. Samples from rocks. residential and light industry. high density. Station 7 Site; Industry: Population: rocky headland. Samples from rocks. outer city residential, paper mill, pine forests, and some agriculture upstream. medium density. Station 8 Site; Industry: Population: mangrove trees in sandy sill. Samples from mangrove roots. dairying, pine forests. medium- to low-density rural; one town, Caboolture, upstream. Results and Discussion Residue levels of DDT. DDD, DDE, and dieldrin calculated on a whole oyster-meat basis from eight samplings at each station are given in Table 2. DDT and dieldrin averages for the period of the survey are shown in graphic form in Figure 2. Vol. 8, No. 3, December 1974 163 ^ DDT & metabolites Q dieldr.n IILifL ^n FIGURE 2. Average pesticide residue levels at Aiisiralmu oyster sampliiti; stations l-H, 1970-72 I Itil'RI V Rainjall and total DDT levels at Australian , 1^)70-72 Highest individiuil residues, 0.94 ppm DDT. 0.5 1 ppm DDD, 0.20 ppm DDE. and 0.34 ppm dieldrin. were obtained in July 1970 at the Brisbane River Sta- tion, in whose catchment area lies the city of Brisbane, population 700.000. .Second-highest levels were from station 3. which adjoins an area of land used intensively for vegetable and fruit production. Most other stations lie downstream from regions with considerably lower levels of human activity. Stations 2 and 7 had levels which were somewhat unexpectedly low. The river adjoining station 2 passes through a cane-growing area where dieldrin is used to control root pests. Station 7 lies at the mouth of a stream on which a paper mill and pine forests are located. Residue levels varied with the seasons. Two factors most likely to influence these variations are tempera- ture and rainfall. Mean shade temperatures in this region range from about 26'C in December and Jan- uary to about 16°C in June and July. A decrease in the pumping rate of the oyster in winter months would tend to lower both the rate of intake of material from the water and the rate of its elimination. Whether this would result in a net loss or gain in chlorinated hydro- carbons is not certain. Reports commenting on seasonal variations do so mainly in terms of rainfall changes (/-.?). In the resion where this survey was conducted, the wet season oc- curs in the summer. December to March, although the winter is comparatively dry. Figures 3 and 4 show- variation in rainfall and the fluctuation in total DDT group levels for stations 3 and 5. It is clear that residue maxima do not coincide with nor immediately follow periods of high rainfall. Other stations, while exhibiting irregularities in some eases, follow this general pattern. This phase dilference between rainfall maximum and residue level maximum may be explained in several FIGURE 4. Rainfall and total DDT levels at Australian oyster saniplini: station 5. 1970-72 ways. Rowe et al. interpret an increase in estuarine oysters near New Orleans as a result of an immediately preceding increase in runofT following heavy rains (3). In the present survey the rainfall maximum preceded the residue maximum by 5 to 11 months, generally with a dry period in between. Thus it is difficult to reconcile these data with the interpretation given below. .■\ general similarity in the pattern of residue vari- ations lound at most sampling stations suggests an alternative explanation. It is possible that the main source ol residues in the region under investigation is the Brisbane River (station 5). and that distribution throughout the bay is efTected by tidal diffusion. However, this interpretation runs into at least two ditficulties. The greatest similarity in the pattern of res- idue variations exists between station 5 and the streams to the soLith (stations 1-4); yet water from the Brisbane River flows predominantly northward. A second prob- 164 Pesticides Monitoring Journal TABLE 2. Chlorinated hydrocarbon levels in oysters (Crassostrea commercialis), Moreion Bay, Queensland, Australia Sampling Sampling Date Residue . ppm Station DDE DDD DDT Dieldrin 1 4-2-70 0.0. A. 1969. The significance of DDT residues in estuarine fauna. In M. W. Miller and G. G. Berg, cds.. Chemical Fallout. Current Research on Persistent Pesticides. Charles C. Thomas Publisher. Sprincfield, 111. 20.^-220. (19) Sinuno. D. R., A. ]. Wil.wn. Jr.. and R. R. Blacknuui. 1970. Localization of DDT in the body organs of pink and white shrimp. Bull. Environ. Contam. Toxicol. 5(4):333-34(). (20) Odiim. W. £., G. A/. WoodwcU. and C. F. Wiir.uer. 1969. DDT residues absorbed from organic detritus by fiddler crabs. Science 164(3879) :576-577. (21) I'clrocelli. S. R.. A. R. Hank.s, and J. W. Anderson. 1973. Uptake and accimiiilation of an organochlorine insecticide (dieldrin) hy an estuarine mollusc, Rangia ciineala. Bull. Environ. Contam. Toxicol. 10(.'i):315- 320. (22) Jensen. S.. A. G. Johnels. M. Olsson, and G. Otler- lind. 1969. DDT and PCB in marine animals from Swedish waters. Nature 224( 52 I 6 ) : 247-2.'50. (2.i') Johnson, B. T.. C. R. Saunders, H. O. .Sanders, and R. S. Campbell. 1971. Biological magnification and degradation of DDT and aldrin by freshwater inverte- brates. J. Fish. Res. Bd. Can. 28(5 ) :705-7()9. (24) Metcalf, R. L.. G. K. Sanfflw, and I. P. Kapoor. 1971. Model ecosystem for the evaluation of pesticide bio- degradability and ecological magnification. Environ. Sci. Technol. 5( 8 ) :709-713. t:.'^) Petrocelli, S. R.. J. W . Anderson, and A. R. Hanks. Biological magnification of dieldrin in a two-parl food chain. Paper presented at the 65th Annual Meet- ing of the National Shellfisheries Assoc. June 24-28, 1973. New Orleans, La, (26) Reinerl. R. E. 1972. Accimiulation of dieldrin in an alga (Seenedesmiis ohliqniis), Daplinia magna, and the guppy iPoecilia relieulata). J. Fish. Res, Bd. Can. 29( I0):1413-1418. (27) Woodwell, G. A/., C. F. Wiirsler, and P. A. I.wacson. 1967. DDT residues in an east coast estuary: a case of biological concentration of a persistent insecticide. Science 156(37761:821-824. ( 2.S ) U.S. Department of Health, Education, and Welfare — Food and Drt/g Admhnstration. 1969. Pesticide ana- lytical manual. Vol. 1. (29) Darnell. R. M. 196S. Food habits of fishes and larger invertebrates of Lake Pontchartrain. Louisiana, an es- tuarine community. Publ. Inst. Mar. Sci. Univ. Tex. 5:353-416. TABLE L Baseline residues (fig/ kg) of DDT and dieldrin in blue crabs, oysters, clams, and brown shrimp, San An- tonio Ba\ System, Tex. — 1972 Sample No.' Wet Weight, n.p'-DUE Blue Crabs 219 98.24 9,47 1,44 April 220 144.86 9 73 ND April 225 = 77.28 28,32 4,22 April 22ft 147,53 22,13 1.38 April 227 111,41 11,14 1.97 April 228 103,91 29,50 6.16 April 229 161 K5 20.20 3.27 April 23(1 94,64 19,40 3,56 April 2.11 191,84 18,54 3 22 April M)^ 159 96 13 80 2,76 June .104 137,81 10,80 TR June .t06 48,75 22 80 ND June .107 66,54 17,60 2,12 June .108 17,39 40,3(1 ND June .109 75,32 12 20 TR June .110 79,63 9,95 ND June 111 29,51 25,80 3,47 June .112 1 1 30 28,00 ND June .11.1 68 85 19,00 2,96 June 114 54,50 13.10 TR June 1.10 91,39 16,30 20,70 June 111 31 53 16,30 ND June .1.12 29 62 51,20 5,41 June .11-1 62,22 13,7(1 2,69 June .166 41,24 21 90 ND Julv ,167 27,03 18 10 2,75 July J68 5 95 ND ND July .169 9,68 ND ND July 170 40,37 19,30 3,50 July 171 23 82 ND 5,76 July 172 1(1 68 14,90 ND July .171 1 1 93 13,10 ND July 174 22 26 27,70 44 60 July 175 17,61 ND 2,64 July 176 29.68 ND 4,41 July .181 146,91 11,70 1 ,30 July 184 100,62 11,40 1 01 July 385 126,48 8,77 ND July 186 11772 9,66 ND July 387 65,13 16,30 ND July 395 148,89 68 20 ND July 396- 296,38 29,80 ND July 397 116,94 3 13 ND July 198 44 39 21 60 ND Julv 410 196 9(1 4.43 ND July 411 168 17 12,10 ND July 412 196 48 10,0(1 ND Julv 411 161 (15 6,54 ND July 518 87 fty 13.60 ND October 519 74 28 6,20 ND October 520 59 01 13 40 ND October 521 77()6 5 90 ND October 522 93,74 18,70 ND October 523 7553 15,80 ND October 584' 64,95 15,00 ND October 585 "■ 53 8S 6 20 ND October 586" 56,74 6 30 ND October 587^ 58 58 55 (10 ND October 588* 93,78 5,2(1 ND October 589" 62 77 4 30 ND October 590" 51 85 15 00 ND October 591- 63,76 8,10 ND October Oysters 213 10 85 29,85 9 28 April 214 5 17 46 5y 1962 April 215 4,16 28.18 22 91 April 216 9,11 27,11 13 38 April 234 7,70 51,87 27,11 April 235 14,10 29,76 13,14 April 236 2 90 72 25 31 9(1 April 237 2 12 70,17 36,08 April 294^ 28 81 ND ND June 295'' 131,73 3,74 ND June 405 3,48 28 50 ND June 406 2 73 28 90 ND June 407 7,14 28 80 ND June 408 9,15 2840 ND June 409 3,96 37,30 ND June 551 8,39 27,00 ND July iContinued next page) 170 Pesticides Monitoring Journal TABLH 1 (com'd). Sample Nn.i Baseline residues lfii;/ki;) of DDT and dieldriii in blue erabs, oysters, clams, and brown shrimp, San Antonio Bay System, Tex. — 1972 Wi:r Wi.iGHi. ;),/)'-DDE Oysters, cont'd Clams Month of Collection 552 7.01 ND 22.20 July 551 7.J8 106.1)0 2(LhO July 554 8.57 24.40 24.20 July 555 7.73 ND ND Julv 556 5.56 ND ND July 557 7.08 ND ND July ^58 2.55 ND ND July S59 3.26 ND ND Julv S6() 4.14 ND ND Julv 592 2.42 ND 26.00 Ocuiber 591 3.21 23.00 ND Ocli>her 594 5.51 25.00 ND Ocmlx-r 595 5.60 48.00 ND October 596 7.53 ND ND Oclober 221 10.89 18.42 13.73 April 222 13.64 22.86 ND April 296 '•' 33.81 21 90 ND June 297" 56.27 18,40 ND June 339 21.51 25,40 ND June 340 23.18 37 (10 6.65 June 341 16.42 36,70 ND June 342 18.68 32,90 3,73 June 343 18.25 42,40 3.75 June 344 20,62 38 70 3.16 June 346 11.75 42,80 9.34 June 347 9,06 59,90 1.07 June 348 10.69 50,20 12.40 June 349 6.28 ND 13.90 June 356 10 85 34,70 3.53 June 357 5.36 ND ND June 358 1 1 .63 54, Kl) 2.50 June 359 10.96 35,80 ND June 360 10.18 ND ND June 541 1- 3.20 ND ND September 542 3.91 ND ND September 543 4.63 ND ND September 544 7.80 ND ND September 561 6.50 36,00 14.00 April 562 7.55 30.00 12.00 April 563 6.36 ND ND April NOTE: Residue totals and averages listed in Table 3. ND =z not delected. TR = trace (< 1.0 yg/kg). 1 Samples represent individual specimens unless indicated otherwise. - Sample taken from Bludworth Island. ^ Sample contained 18.60 ^g kg p,p'-DDE. * Composite ol two crabs. ^'Composite of six crabs. •'Composite of four crabs. Sample No.' Wet Wcicht. /),p'-DDE DiELORIN Clams, cont'd Brown Shrimp " Composite of three crabs. ■■ Composite of ten small oysters. ■' Composite of ten large oysters. "■Composite of ten small clams. " Composite of ten large clams. '- Sample taken from Sundown Bay. ' ' Sample contained 33,00 ^y kg p,p' " Composite of five clams. '"■ Sample contained 47.00 ^g kg p.p' DDE. ■DDE. Month of Collection 564 6,22 41,00 ND April 565 ' ■ 7,K3 15-0(1 ND April 566 7,52 ND ND April 567 699 10,00 ND April 568 9,20 ND ND April 569 7,83 ND ND April 570 7(12 ND 5 10 April 571 ■' 18,67 ND 2,00 April 598 6 10 ND ND October 599 4 71 18,00 ND October 600 4,58 ND 18,00 October 601 3.65 ND ND October 602 2.82 79.00 ND October 603 0,74 20 (10 ND October 605 8 12 ND ND October 606 '•■ 5 92 16(10 ND October 607 12,33 16,00 ND October 240 6.48 ND 6.70 April 241 7.82 ND 4.51 April 242 8.16 ND 4.38 April 249 6.71 ND 8,71 April 250 9.44 ND ND April 251 5.64 ND 6,5(1 April 252 10.15 ND ND April 254 5.73 ND 1 1 .36 April 256 887 ND ND April 257 6 59 ND ND April 258 12-4(1 ND ND April 259 6-74 ND ND April 266 4.23 ND ND May 267 3.80 ND ND May 268 6.20 ND ND May 269 4.81 ND ND May 270 3.53 ND ND May 271 5.98 ND ND May 272 5.08 ND ND May 273 10.65 ND ND May 402 5.60 15.70 ND July 403 5.52 14.70 ND July 404 5.58 13.70 ND July Vol. 8, No. 3, December 1974 171 TABLE Baseline n'sitlitc.s (tjg/ki;) of DDT and dicldrin in niisccllanciiiis species, San Antonio liav Svstcm. Tex. — 1972 Sample Month of No. Organism i l.ncAiioN - W'l 1 \Vl K.lll. 1, /..r-DDE Dll 1 DRIN Collection ::? AikIjoci iniululli Bliidworlh Island .12.SI> 52,61 5, IS April 2:4 TLii;eliis i>lvbeiu\ Bliiduorth Island 29X4 5.67 2 68 April 238 Parulicljlhvs lelhostiiima Refuge 14.72 13.32 ND April 239 Paraltchtliyi lethosttKnid Refuge 14,78 14.21 ND April 435 Mereemtria ciimpechit't}\i\ Hvnes Bav 75-95 ND ND July 436 Mercenana ccintpec/iu'i!\n Hvnes Bav 34,28 ND ND July 437 hlercenm ia campeclueti\i\ Hvnes Bay 55,66 ND ND July 438 M ercenm ia eiinipechien\is Hynes Bay 25,41 ND ND July 439 Merceiuina CiiinpechienM\ Hvnes Bay 42 58 ND ND July 450 TaaeUts pleheius Refuge 4.75 ND ND July 451 Tiif^eltfs pleheius Refuge 8,07 ND ND July 452 Maconiii conslricta Refuge 2,8.1 ND ND July 526 Sparlinti spairinae Dunham Bay 57,43 2.20 ND August 527 Sparlina spartinae Muslang Lake 57,25 ND ND August 528 Sparlma spurtinae Dunliam Bav 49,11 ND ND 533 Mercenaria campechiensis Hvnes Bav 95,26 ND 1.30 October 534 Mercenana campecluensis Hvnes Bav 135 69 ND 1.20 October Merceiiiiria catitpechiensis Hvnes Bay 1(19,45 ND 1.40 536 Mercenana campecluen\i.s Hynes Bay 155.47 ND 1.00 October 537 Mercenaria campec/iiensii Hynes Bay 20.43 ND ND 538 Mercenaria canipechien},i-v Hynes Bav 11.52 ND ND 539 Mercenana campechiensis Hynes Bav 17.95 ND ND 540 Tagelas plebeian Sundown Bay 4 24 ND ND September ' See text for cammon names -Compare map. Fig. 2 TABLE 3. Summary of insecticide residues detected in biological samples, San Antonio Bay System, Tex. 1972 No. Organisms Analyzed No. Organisms Containing Insecticide Residues and Percent Total Analyzed Species P.P'-DDE DiELDRIN Both Blue crabs Oysters Clams Shrimp Miscellaneous 81 48 65 23 23 76 (94%) 29 (60%) 44 (68%) 3 (13%) 7 (30% ) 26 (32%) 12 (25%) 20 (31%) 6 (26%) 6 (26%) 23 (28%. ) 10 (21%) 12 (18%) 0 ( 0% ) 2 ( 9% ) Total 240 159(66%) 70 (29%) 47(20%) 172 Pesticides Monitoring Journal Residues of 2,4-D in Pond Waters, Mud, and Fish, 1971 ^'^ Donald P. Schultz and Paul D. Harman ABSTRACT Nine ponds in Florida, Georgia, and Missouri were treated in 1971 with the dimethylamine salt of 2,4-Dichlorophe- noxyacetic acid (DMA-2,4-D) to determine residue levels of the herbicide in water, bottom mud, and fish as a function of application rate, time, substrate, and geographic location. The acid equivalent of 2,4-D was applied at treatment rates of 2.24, 4.48. or 8.96 kg/ha. Samples were taken up to 147 days after treatment. Residues of 2,4-D in pond waters declined from a maximum of 0.345 and 0.692 mg/liter in Florida and Georgia, respec- tively, to less than 0.005 mg/liter 28 days after treatment, and from 0.630 mg/liter to less than 0.005 mg/liter 56 days after treatment in Missouri pond waters. Residues in mud from the Florida and Georgia ponds never exceeded 0.05 mg/kg and had declined to trace or non- detectable levels 56 days after treatment. The highest residue found was 0.170 mg/kg from the first- and third-day samples of mud in the most heavily treated Missouri pond. In mud from one Missouri pond, residues were detected as late as 28 days after treatment; no residues occurred in any ponds after that. No evidence of fish kill was found in any ponds treated with DMA-2,4-D. Of 307 fish samples analyzed, 45 con- tained detectable residues of 2,4-D. Residue levels ranged from slightly more than 1.0 mg/kg to less than 0.010 mg/kg. Introduction Proliferation of obnoxious aquatic plants is presently controlled by mechanical, physical, biological, or chemical means, or by combinations of these methods. Formulations of 2,4-D are used most widely to control plants such as the waterhyacinth (Eiclwrnia crassipe.^) and eurasian watermilfoil {Myriophyllum spicatuin). 1 Southeastern Fish Control Laboratory, Fish and Wildlife Service, U.S. Department of the Interior, Warm Springs, Ga. 31830. ^ Supported in part by the U.S. Army Corps of Engineers. In 1966 the Tennessee Valley Authority applied 888 tons of the butoxyethanol ester of 2,4-D to 8,000 acres of watermilfoil in several reservoirs (/). Over 18,000 surface acres of Nickajack and Guntersville Reservoirs on the Tennessee River were treated with about 170.000 gallons of the dimethylamine salt of 2,4-D (DMA-2,4-D) April — June 1969, to control watermilfoil (2). Objectives of the present study conducted in 1971 were to determine the residue levels and rate of dissipa- tion of DMA-2,4-D in fish, water, and bottom mud. These data are required to register the product for use in aquatic plant control and to assist in establishing residue tolerances for DMA-2,4-D in fish and domestic water. Also, by using ponds in Florida, Georgia, and Missouri, authors hoped to determine whether different physical and chemical characteristics of the aquatic environment affect the uptake and dissipation of the herbicide. A third objective was to assess the efficacy of the herbicide on minor infestations of waterhyacinth in Florida and Georgia ponds. Methods and Materials SITE SELECTION Three ponds were located near Crystal River, Fla. Willow Pond contained some aquatic plants including Hydrocotyle sp., cattail {Typha sp.), waterlettuce (Pistia stratiotes), widgeongrass (Ruppia tnaritima), and Sagittaria sp. in about 5 percent of its total area. One bank was heavily covered with willow (Salix sp.). Shelter Pond contained a dense stand of Char a sp., which covered about one-third of the bottom, and Hydrilla verticillata. which covered about 50 percent of the pond area at the beginning of the experiment. The third pond, designated 11-T, contained small amounts of widgeongrass, Sagittaria sp., and Hydroco- tyle sp. Initially, about 30 percent of its area contained Vol. 8, No. 3, December 1974 173 Hydrilla sp. All three ponds were treated with rotenone in May 1971 and restocked in June 1971 with large- mouth bass {Microptenis salmoides) . channel catfish (Ictaliinis piinctatiis) . bluegill (Lepomis macrochirus) . and redear sunfish {Lepomis niicrolophiis) . One month prior to treatment the three ponds were stocked with waterhyacinth so that it would cover 5-10 percent of surface area of each pond at the time of treatment. A second study area of four ponds was located within a 10-mile radius of Warm Springs, Ga., on the Piedmont Plateau and was comprised of four ponds which con- tained little, if any, submersed vegetation. Several of the ponds had small stands (less than 5 percent coverage) of cattail. Because the ponds contained established fish populations of the desired species, only enough fish were added to ensure an adequate number for the ex- periment, Waterhyacinths were transported from Flor- ida to stock the ponds; their total coverage did not exceed 5 percent of the surface area of each pond. The ponds were designated 0, 2, 4, and 8 in accordance with the amount of herbicide applied. Four ponds representing a third ecological and geo- graphical type were located at the USDI Fish and Wild- life Service, Fish-Pesticide Research Laboratory, Co- lumbia, Mo, Pond 9 contained no emersed or submersed macrophytes be;ause of its prior use as a crayfish holding pond. Pond 15, the control, contained a com- mon grass (Arena sp,) which had established itself when the pond was dry. About 5 percent of the surface area of Pond 28 contained cattails. There was also a 20-30 percent infestation of an unidentified filamentous alga. About 10 percent of the surface area of Pond 26 contained smartweed {Polygonum sp,). Physical characteristics of the ponds used in the ex- periment are listed in Table 1. TABLE 1, Physical characteristics of ponds in Florida, Georgia, and Missouri, and initial 2,4-D concentrations, 1971 Surface Volume. AREA. ACRES ACRE- FT Depth, ft 2,4-D, Pond Average Maximum MG/L Fla-Willow 0.60 2.54 4.23 7.0 0.174 Fla-Shelter 0.43 1.45 3.37 6.0 0.436 Fla-ll-T 0.80 3.12 3.90 8.0 0.801 Ga-0 1.30 7.30 5.60 11.0 0.000 Ga-2 0.60 2.65 4.40 8.0 0.166 Ga-4 0.90 2.70 3.00 7.0 0.490 Ga-8 0.86 2.95 3,40 7.0 0.857 Mo-15 0.073 0.166 2.26 4.5 0.000 Mo-9 0.073 0.166 2.26 4.5 0.444 Mo-28 0.162 0.368 2.26 4.0 1.002 Mo-26 0.140 0.451 3.20 4.5 1.631 NOTE: Florida ronds had sandy boltoms covered with a muck layer 1-3 inches deep, Georgia ponds had red clay bottoms charac- teristic of the Piedmont Plateau, and Missouri ponds had bot- toms of heavy colloidal clay. TRF.ATMENT DMA-2,4-D was applied to nine ponds at 2,24, 4,48, or 8.96 kg per hectare (kg/ha,) (2, 4, or 8 lb/acre) using a commercial formulation of herbicide (Weedar 64, Amchem Products, Ambler, Pa.) which contained 4 lb 2,4-D acid equivalent (a.e.) per gallon. All dilu- tions were made with water and no adjuvants were used. The Florida ponds were sprayed July 12, 1971, and the Georgia and Missouri ponds were sprayed July 26, 1971, SAMPLING Samples of fish, water, and mud were taken at 0. 1, 3, 7, 14, 28, 56, 84 or 86, 112, and 140 or 147 days after treatment. Fish were placed in live-cages in the Florida ponds for first- and third-day samples. There- after, fish were collected by hook and line, seine, or setline; they were weighed, measured, wrapjjed in aluminum foil, bagged, and frozen on dry ice. Water samples were taken with a 2-liter Kemmerer water bottle: they were composites of samples from areas whose depths varied from shallow ( 1 foot) to medium (2-3 feet) to deep (4 feet or more). The pH of water samples was determined either colorimetrically or with a pH meter. Water for residue analysis was placed in quart jars and acidified to a pH of less than 2 with concentrated sulfuric acid. Jars were capped with aluminum foil and sealed with screw caps. Mud samples were taken with an Ekman dredge from shallow, medium, and deep sites. The three samples were composited for residue analysis, placed in plastic bags, and frozen on dry ice until analysis, A nalytical Procedures Acidified water samples were extracted with chloro- form which was subsequently evaporated. The 2,4-D residues were derivatized with diazomethane (5) for quantification by gas-liquid chromatography (GLC). Mud samples were acidified and extracted in a blender with a mixture of acetone, petroleum ether, and diethyl ether. The slurry was filtered and extracted twice with base (4), TTie basic solution was acidified and ex- tracted with chloroform and the chloroform solution was treated as above. Fish samples were ground with dry ice (5) and the resultant powder was packed in a column and extracted with acidified methanol (6). The methanol eluate was added to water, the pH v/as adjusted to 1,5, and the solution was extracted with diethyl ether. The diethyl ether extract was treated as described earlier for chloroform extracts, Esterified 2,4-D residues were cleaned up on a column of silica gel. The gas chromatograph used to quantify 2,4-D residues was equipped with an electrolytic conductivity detector (7), Recovery values from spiked water, mud, and fish samples were 97,5 ± 2.5, 90 ± 2.5, and 90 ± 2,5, respectively. 174 Pesticides Monitoring Journal All solvents used for extractions were glass-distilled (Biirdick and Jackson, Muskegon. Mich.). All other chemicals used were analytical reagent grade. Residue levels are given in terms of the methyl ester of 2.4-D and were not corrected for percent recovery. The detection limit of the 2.4-D methyl ester was 1.25 ng; 15 ng registered a 50 percent deflection on the recorder. Detection limits were 0.01 mg/kg, 0.005 mg/kg. and 0.001 mg/liter for fish. mud. and water samples, respectively. Confirmatory analyses of the 2,4-D methyl ester standard, spiked samples, and periodic samples were conducted by electron-capture detection on a Beckman GC-4 gas chromatograph. Detailed procedures for extraction of 2,4-D residues from water, mud. and fish samples, as well as the derivatization and gas chromatographic procedures, have been described by Schultz and Whitney (7). Results WATER In five of the nine treated ponds, highest residue levels were detected the third day after treatment. Presumably it took this length of time for the herbicide to be thoroughly dispersed in the water. The highest level of 2,4-D residues in Florida pond water was 0.345 mg/ liter; it was found 3 days after herbicide application to Pond 11-T, which was treated at 8.96 kg/ha. (8 lb/acre). Residue levels of 2,4-D in Florida ponds decreased to no more than 0.005 mg/liter within 14 days after treatment. Thereafter, only trace amounts (less than 0.005 mg/liter) or no residues were detected (Table 2). In Georgia pond water the highest detectable residue was 0.692 mg/liter found in Pond 8 three days after treatment at 8.96 kg/ ha. Residue levels of 2,4-D in these waters decreased to trace amounts (less than 0.005 mg/liter) within 28 days after treatment; no residues were detected thereafter (Table 2). TABLE 2. Residues of DMA-2,4-D in water (mg/L) and mud (mg/kg) samples from ponds treated witli 2,4-D, 1971 Treatment, pond, and SAMPLE Days after treatment 28 56 84/86 1 FLORIDA GEORGIA MISSOURI 112 140/147 2 2.24 kg/ha. (Willow Pond) water mud ND ND 0.125 TR 0.025 TR TR 0.005 TR TR TR ND TR ND TR ND ND ND ND ND 4.48 kg/ha. (Shelter Pond) water mud ND ND 0.155 0.014 0,172 0.014 0.048 0,010 0.005 0.010 TR 0.007 ND ND TR TR ND ND ND ND 8.96 kg/ha. (II-T) water mud ND ND 0.312 0.033 0.345 0.046 0.025 0.008 0.005 0.013 TR ND ND TR TR TR ND TR ND ND No herbicide (Pond 0) water mud ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 2.24 kg/ha. (Pond 2) water mud ND ND 0.025 0.018 0.087 0.008 0.059 0,010 0.027 0.006 TR TR ND ND ND ND ND ND ND ND 4.48 kg/ha. (Pond 4) water mud ND ND 0,233 0.024 0,390 O.OIS 0,400 0.050 TR TR ND ND ND ND ND ND TR ND ND 8.96 kg/ha. (Pond 8) water mud ND ND 0,617 0.026 0.692 0.040 0,395 0.042 0.008 TR TR ND ND ND ND ND ND ND ND ND No herbicide (Pond 15) water mud ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 2.24 kg/ha. (Pond 9) water mud ND ND 0.104 ND 0.108 TR 0.102 0.009 0.050 ND 0.017 ND ND ND ND ND ND ND ND ND 4.48 kg/ha. (Pond 28) water mud ND 3 0.160 0.011 0.256 0,012 0.250 0.005 0.480 TR 0.150 ND ND ND ND ND ND ND ND ND 8.96 kg/ha. (Pond 26) water mud ND s 0,580 0.170 0.420 0.170 0.326 0.091 0.630 0.068 0.135 0.005 ND ND ND ND ND ND ND ND NOTE: ND = not detectable; TR = trace (less than 0.005 mg/L or kg). 1 In Florida and Georgia the seventh posttreatmem sampling was made 84 days after application; in Missouri it was made 86 days after appli- cation. = In Florida and Georgia the last sampling was made 140 days after application; in Missouri it was made 147 days after application. =* Sample was lost. Vol. 8, No. 3, December 1974 175 The most persistent residues were found in the two most heavily treated Missouri ponds which contained more than 0.1 mg/ liter of 2,4-D 28 days after treatment (Table 2). However, no herbicide was detected 56 days after treatment or thereatter. were in fish from the first-day harvest which had been held in live-cages in the Florida ponds. The 2,4-D residues found in the exposed fish were well below the levels of the dimethylamine salt formulation found to be to.xic to fish (9-11). MUD Highest 2,4-D residues in mud were consistently found in the ponds which had received the heaviest herbicide treatment. The largest 2,4-D residue detected in mud from Florida ponds was 0.046 mg/kg found in Pond 11-T three days after treatment (Table 2). Trace amounts were found in the same pond up to 112 days after treatment. In the Georgia ponds the highest residue found was 0.042 mg/kg in Pond 8 seven days after treatment (Table 2). A residue level of 0.170 mg/kg from Missouri Pond 26 was found at the first- and third-day samplings; no residues were detected in any other Missouri pond 56 days after treatment or there- after (Table 2). FISH Highest residues found in fish were from the first-day harvest of Willow and Shelter Ponds in Florida (Table 3). These fish were kept in live-cages during and after treatment to facilitate sampling; hence they were unable to escape from the applied herbicide {8). No residues were detected in fish from Florida ponds during the third- or seventh-day harvests; however, residues were detected 14 days after treatment. This may have been caused by release of the herbicide from decaying vegetation. Only one fish from the Florida ponds con- tained a detectable residue after 14 days. It was one of two largemouth bass from Pond 11-T, which had been treated at 8.96 kg/ha.; the residue was a trace amount (0.005 mg/kg). The highest 2,4-D residue found in fish from the Georgia ponds was 0.075 mg/kg in one of three blue- gills harvested 14 days after pesticide application at 8.96 kg/ha. (Table 3). No detectable residues were found in any fish from the Georgia ponds at the third- or seventh-day harvests; this parallels results of Florida pond samples. With one exception, the last detectable residues were found 28 days after treatment. The exception was a bluegill from Pond 8: it contained less than 0.005 mg/kg 84 days after treatment. No residues of 2,4-D were detected in fish from the Missouri ponds until 28 days after treatment (Table 3). At that time all five bluegills sampled and one of five largemouth bass sampled had less than 0.005 mg/kg. No other fish from the Missouri ponds contained detectable residues of 2.4-D. None of the fish from control ponds contained de- tectable residues of 2,4-D. The four highest residues WATERHYACINTH CONTROL The effect of the 2,4-D application on waterhyacinth in the Florida and Georgia ponds was assessed by visual observation. Nearly all the waterhyacinth was brown and decomposing 7 days after spraying. Approxi- mately 98 percent of the plants had been killed by the herbicide application; no difference in kill was noted among the different treatment levels. Spot retreatment was needed in some areas of all ponds to prevent reinfestation from the estimated 2 percent of the waterhyacinth which survived the initial application. TABLE 3. Residues of DMA-2,4-D in fish from ponds treated with 2,4-D, 1971 Species Days AFTER TREATMENT Residues (mo/kg) in fish treated at— AND FISH NUMBER 0 kg/ha. 2.24 kg/ha. 4.48 kg/ha. 8.96 kg/ha. FLORIDA LMB-1 0 ND LMB-2 0 ND CCF 0 ND LMB-1 0.080 0.048 TR LMB-2 ND LMB-3 0.008 CCF-1 1 .075 0.340 ND CCF-2 ND CCF-3 ND BLG-1 0.024 0.420 0.010 BLG-2 TR BLG-3 0.012 LMB ND ND ND CCF ND ND ND BLG ND TR ND LMB 7 ND ND ND ND CCF 7 ND ND ND BLG 7 ND ND ND ND LMB-1 14 ND 0.036 ND 0.043 LMB-2 14 0.031 ND LMB-3 14 TR CCF-1 14 0.029 0.050 0.024 CCF-2 14 0.032 0.012 0.102 CCF-3 14 0.012 0.039 0.094 BLG-1 14 ND ND 0.018 ND BLG-2 14 ND ND 0.008 ND LMB-1 28 ND ND ND LMB-2 28 ND ND TR LMB-3 28 ND CCF-1 28 ND ND CCF-2 28 ND ND ND CCF-3 28 ND ND ND BLG 28 ND ND ND LMB 56 ND ND ND CCF 56 ND ND ND BLG 56 ND ND ND LMB 84 ND ND CCF 84 ND ND BLG 84 ND ND LMB 112 ND CCF 112 ND ND ND BLG 112 ND ND ND LMB 140 ND ND ND CCF 140 ND ND BLG 140 ND ND (Continued next page) 176 Pesticides Monitoring Journal TABLE 3 (cont'd). RcsiJuc.\ of DMA-2,4-D in fi.\h from ponds ircalcd with 2,4-D, 1971 Species Days AFTER TREATMENT Residues (mg/kg) in fish treated at — AND FISH NUMBER 0 kg/ha. 2.24 kg/ha. 4.48 kg/ha. 8.96 kg/ha. Species Days AFIER TREATMENT Residues (mc/kg) in fish treated at— AND FISH NUMBER 0 kg/ha. 2.24 kg/ha. 4.48 kg/ha. 8.96 kg/ha. GEORGIA LMB 0 ND CCF-1 0 ND CCF-2 0 ND BLG 0 ND LMB ND 0.014 0.022 CCF ND ND 0.043 0.008 BLG ND ND 0.024 0.044 LMB-1 ND ND ND ND LMB-2 ND ND LMB-3 ND CCF-1 ND ND ND ND CCF-2 ND ND ND ND BLG ND ND ND LMB-1 ND ND ND ND LMB-2 ND ND LMB-3 ND CCF-1 ND ND ND ND CCF-2 ND BLG ND ND ND ND LMB-1 ND ND ND LMB-2 ND ND LMB-3 ND ND CCF- 1 ND ND 0.012 CCF-2 ND CCF-3 ND BLG-1 ND ND 0.075 BLG-2 ND ND BLG-3 ND ND LMB-1 28 ND ND ND O.OIO LMB-2 28 ND ND ND 0.005 LMB-3 28 ND ND LMB^ 28 ND CCF-1 28 ND ND ND ND CCF-2 28 ND ND CCF-3 28 ND ND BLG 28 ND ND ND ND LMB-1 56 ND ND ND ND LMB-2 56 ND ND ND CCF 56 ND ND ND ND BLG 56 ND ND ND ND LMB 84 ND ND ND ND CCF 84 ND ND ND BLG-1 84 ND ND BLG-2 84 TR LMB-1 112 ND ND ND ND LMB-2 112 ND CCF 112 ND ND ND ND BLG 112 ND ND ND ND LMB-1 140 ND ND ND ND LMB-2 140 ND ND LMB-3 140 ND LMB^ 140 ND LMB-5 140 ND CCF-1 140 ND ND ND CCF-2 140 ND ND BLG-1 140 ND ND ND BLG-2 140 ND BLG-3 140 ND BLG-4 140 ND BLG-5 140 ND MISSOURI CCF 1 ND ND BLG LMB 1 1 ND ND ND ND CCF 7 ND BLG 7 ND LMB 7 ND CCF 14 ND BLG 14 ND LMB CCF-1 14 14 ND ND CCF-2 14 ND CCF-3 CCF-4 14 14 ND ND CCF-5 14 ND BLG-1 14 ND BLG-2 14 ND BLG-3 14 ND MISSOURI, cont'd BLG-4 14 ND BLG-5 14 ND LMB-1 14 ND LMB-2 14 ND LMB-3 14 ND LMB^ 14 ND LMB-5 14 ND CCF 28 ND BLG 28 ND LMB 28 ND CCF-1 28 ND CCF-2 28 ND CCF-3 28 ND CCF-4 28 ND CCF-5 28 ND BLG-1 28 TR BLG-2 28 TR BLG-3 28 TR BLG-4 28 TR BLG-5 28 TR LMB-1 28 ND LMB-2 28 TR LMB-3 28 ND LMB-4 28 ND LMB-5 28 ND CCF 56 ND BLG 56 ND LMB 56 ND LMB = largemouth bass, CCF = channel catfish, BLG = bluegills. ND = not detectable. TR = trace (less than 0.005 mg/kg). Blank = no sample analyzed. Discussion WATER Residues in Missouri pond waters were more persistent than were Florida and Georgia residues. This may have been caused partly by cooler water temperatures in the Missouri ponds which may have slowed the biological decomposition. For example, the median water tem- perature (shallow, medium, deep) in the three Missouri ponds averaged 26,6°C from the day of treatment to 28 days thereafter, whereas average temperatures in the Georgia and Florida ponds were 28.0°C and 30.9°C, respectively. Also, the Missouri ponds had been sprayed on the assumption that they were either 0.1 or 0.25 acres in area. However, when morphometric measurements were made, it was found that their areas in acres were as follow: Pond 9, 0.073; Pond 28, 0.162; and Pond 26, 0.140. Hence the initial concentration of 2,4-D (mg 2,4-D/liter water) in Missouri ponds was almost twice that in Florida and Georgia ponds (Table 1). Residual persistence of the herbicide in Missouri ponds may also be attributed to direct application of the herbicide to the water surface. Because no waterhya- cinth was present, there was little surface biomass to absorb or adsorb and degrade the chemical. Vol. 8, No. 3. December 1974 177 Wojtalik et al. (2) reported residues greater than 0.02 mg/liter at only 2 of 19 stations 4 weeks after treatment at 20 or 40 lb a.e./acre DMA-2,4-D. Aly and Faust (12) reported a photodecomposition loss of 50 percent of a sodium salt of 2,4-D in 50 minutes at pH 7. Hence the disappearance of DMA-2,4-D from water is rapid compared to the herbicide dichlobenil, 2.6- Dichlorobenzonitriie, which had a reported residue of 0.05 mg''liter 85 days after treatment at 0.55 mg/liter. and the herbicide fenac. 2,3,6-Trichlorophen>lacetic acid, which had a reported residue of 0.77 mg/liter 85 days after treatment at 1.56 mg/liter (13). The dissipation of 2,4-D from mud was quite rapid compared to dissipation of fenac and dichlobenil, which have been reported at 0.06 and 0.12 mg/kg. respective- ly, 160 days after treatment (!3). Presumably, the dissipation of 2,4-D from mud was caused in great measure by microbiological degradation. At least 1 1 species of bacteria and two actinomycctes are known to degrade 2,4-D (14). FISH Chemicals can impart an off-taste or even toxicity to fish consumed by humans; hence it is imperative that residue levels in fish of any pesticide applied to aquatic environments be as low as possible. At this writing, no tolerance levels have been set for residues of DMA-2, 4-D in fish. Only 7 percent of the fish analyzed 28 days or more after treatment contained detectable 2,4-D residues and only 1 percent (one fish) of those analyzed 56 days or more after treatment contained detectable residues (Table 3). Thus if tolerance levels are based on the level of parent compound only, it would appear that fish could be consumed I month after treatment. However, degradation products were present in fish for as long as 84 days after treatment in plastic pools ( //). Hence the presence of these unidentified products also should be considered. Little, if anything, is presently known about the direct effects of herbicides on fish reproduction. However, reproduction of bluegills occurred in two Florida ponds. Willow and 11-T. during the experiment. Growth and development of the fry appeared to be normal. At no time was there any evidence of fish mortalitv caused by the herbicidal application. Pesticides usually biomagnify through the food chain. Authors found that fish absorbed little 2,4-D when fed radio-labelled herbicide (/7). Also, Wojtalik et al. (2) reported no harmful effects or accumulation in zooplankton, phytoplankton, or macroinvertebrates in water treated at 20 or 40 lb a.e./acre. Hence there is little danger of biomagnification of 2. 4-D in contrast to the chlorinated hydrocarbon pesticides (/5). How- ever, when fish were exposed to '^C-DMA-2,4-D in water, radioactive compounds were ubiquitous in all tissues examined (11). Therefore, the identity and potential toxicity of these compounds must not be overlooked. It also has been reported that although fish absorbed more 2,4-D at pH 6 than at pH 9, they retained more of the parent compound at a basic pH (//). Thus, aquatic applications of 2,4-D probably should be avoided under conditions of high pH. Although fish can be killed by high concentrations of DMA-2,4-D, the reported TL,„ at 96 hours (//) is high enough that even a tenfold error in application rate would not decimate a fish population. In the same study, death of fish from the herbicide was found not to result from a single, specific factor such as a carcinogenic effect; it undoubtedly resulted from a plethora of effects on carbohydrate metabolism. On the basis of this study, then, waterhyacinth can be controlled by judicious application of DMA-2, 4-D at recommended rates without concomitantly high residue levels in fish, water, or hydrosol. A cknowledgment Authors gratefully acknowledge the help of the follow- ing people from the Fish-Pesticide Research Laboratory, Columbi.T, Mo.: Betty Barnum, James Huckins, and Dave Zumwalt. LITERATURE CITED (1) Smith, G. E.. and B. G. Isom. 1967. Investigations of effects of large-scale applications of 2. 4-D on aquatic fauna and water qualitv. Pestic. Monit. J. 1(1):16-21. (2) Wojtalik. T. A.. T. F. Hall, and L. L. Hill. 1971. Monitoring ecological conditions associated with wide- scale applications of DMA-2,4-D to aquatic environ- ments. Pestic. Monit. J. 4(4 ): 184-203. {3] Howard. S. F., and G. Yip. 1971. Diazomethane methylation of a mixture of chlorophenoxy acids and dinilrophcnols. J. Ass. Offic. Anal. Chcm. 54(4):97()- 974. (4) Woodhain. D. W., W. G. Miichcll. C. D. Loffis, and C. W. Collier. 1971. An improved gas chromatogra- phic method for the analysis of 2,4-D free acid in soil. J. Agr. Food Chem. |4( 1 ); 186-I!. u. E c ■a > c dj '■J >'p > ti „ aj c ■ - TJ C c -a u W «< o; C a. Z- o .5 =5 I 5 a 5; o ^ c m O o zg Z'. +1 +1 +1 +1 +1 o o o +1 +1 41 + 1 +1 +1 +1 c o o o C ± '^ z ceo c c c +; +1 +1 +1 +' + ! +1 coo c § □ i c c •SEE 182 Pesticides Monitoring Journal Gas chromatographic analysis indicated that lindane, dieldrin, DDE, DDD. and DDT were present in both feed samples and fat from slaughtered animals (Table 2). Average residues in fat were lindane. 0.026 ppm; DDE. 0.261 ppm: dieldrin. 0.030 ppm; DDD. 0.101 ppm; and DDT, 0.292 ppm. Heptachlor epoxide resi- dues were not detected in any samples analyzed. Values in Table 2 represent means and standard deviations (uncorrected for recovery) calculated from 10 fat samples and 5 feed samples in each group. Residues reported herein are based on adipose tissue as removed from the animals. Such tissue contains 80-85 percent extractable fat. No evidence of toxicity was observed in any of the cattle; this was expected, considering the very low levels of pesticide consumed. However, observations were made during the test for possible enhancement of toxicity from the addition of ammonium salts to the diet. All animals gained weight normally during the test and showed no evidence of toxic conditions at slaughter. With the exception of the insecticide dieldrin. all resi- dues found in fat samples were much higher than resi- dues observed in feed. Magnification of this type is well documented and is to be expected. Dieldrin residues present in the fat of each of the animal groups very closely approximate levels found in the diet. The Dun- can Multiple Range Test (14). used to determine sig- nificance of differences in residues resulting from the test diets, indicated that with the exception of lindane residues in the fat of animals receiving high ammonium sulfate levels, pesticide residue levels were not aff^ected by addition of either ammonium chloride or ammo- nium sulfate to the diet. Fat samples from animals re- ceiving 100 g/head/day ammonium sulfate had lindane residues significantly lower than those from other treated animals or from control animals. Conclusions Low residue levels of five chlorinated insecticides, lin- dane, dieldrin, DDE, DDD, and DDT, were found in feed and fat of west Texas feedlot cattle. Levels in feed were less than 0.1 ppm; those in fat were less than 0.5 ppm. Except for decreased lindane residues in cattle receiving 100 g ammonium sulfate daily, chlorinated pesticide residues were not affected by ammonium chloride or ammonium sulfate when ammonium compounds were used as nitrogen supplements. No ill effects resulting from interaction of pesticides and ammonium compounds were observed in any of the groups. The analytical method was sensitive and efficient for analysis of fat samples for low levels of multiple chlori- nated insecticide residues. A cknowledgments Authors gratefully acknowledge the contributions and cooperation of Dr. George F. Ellis, Jr., Feedlot Man- ager, and Dr. Dale Furr, Livestock Nutritionist, Mid- land, Tex. LITERATURE CITED ( / ) Sania Fe Crop Report. June 1972. (2) Moiibry, R. J.. G. R. Myrdol, and H. P. Jensen. 1967. Chlorinated hydrocarbon residues in or on alfalfa grown in soil with a previous history of aldrin and heptachlor application. Pestle. Monit. J. 1(2):13-14. (.?) Marth. E. H. /9rt5. Residues and some effects of chlo- rinated hydrocarbon insecticides in biological material. Residue Rev. 9:1-89. i.4) Fintayson, D. G., and H. R. McCarthy. 1965. The movement and persistence of insecticides in plant tis- sue. Residue Rev. 9: 1 14-152. (5) Bruce. W. N.. G. C. Decker, and J. G. Wilson. 1966. Relationship of the levels of insecticide contamination of crop seed to their fat content and soil concentra- tions of aldrin, heptachlor and their epoxides. J. Econ. Entomol. 59:179-181. (6) Nen-som, L. D. 1967. Consequences of insecticide use on nontarget organisms. Annu. Rev. Entomol. 12:257- 286. (7) Daniels. N. E. 1968. Insecticidal residues in soil and grain sorghum. Tex. Agr. Exp. Sta. Bull. PR-2531. (5) Du<>f>an. R. £., and H. R. Cook. 1971. National food and feed monitoring program. Pestic. Monit. J. 5(1): 37-43. (9) Crookshank. H. R., F. E. Keating, Earl Burnett, J. H. Jones, and R. D. Davis. 1960. Effect of chemical and enzymatic agents on the formation of urinary calculi in fattening steers. J. Anim. Sci. 19:595-600. (10) Crookshank, H. R. 1970. Effect of ammonium salts on the production of ovine urinary calculi. J. Anim. Sci. 30:1002-1004. (//) Crookshank. H. R., H. E. Smalley, Dale Furr, and George F. Ellis. I97J. Ammonium chloride and am- monium sulfate in cattle feedlot finishing ration. J. Anim. Sci. 36:1007-1009. (12) Kadoiim, A. M. 1967. A rapid micromethod of sam- ple cleanup for gas chromatographic analysis of in- secticidal residues in plant, animal, soil, and surface and ground water extracts. Bull. Environ. Contam. Toxicol. 2(5):264-273. (/.?) Kadoum, A. M. 1968. Application of the rapid mi- cromethod of sample cleanup for gas chromatographic analyses of common organic pesticides in ground water, soil, plant and animal extracts. Bull. Environ. Contam. Toxicol. 3:65-70. (14) Duncan, D. B. 1955. New Multiple Range Test. Bio- metrics 11:1-42. Vol. 8, No. 3, December 1974 183 GENERAL DDT and Dieldrin in Watersheds Draining the Tobacco Belt of Southern Ontario R. Frank.i K. Montgomery,^ H. E. Bniun.i A. H. Berst.a and K. Loftus ■» ABSTRACT Production of flue-cured tobacco in Ontario is concentrated mainly in four watersheds adjacent to Norfolk County on the north shore of Lake Erie, where tobacco is raised on a sandy loam soil using a 2- to 3-year rotation with cereals and corn. In 1961-69 an average amount of 4.4 kg/ha. DDT was used to control soil and foliar insects attacking tobacco. In 1970-71, DDT was restricted to the control of soil insects only; applications averaged 1.7 kg/ ha. Residue data from soil samples collected in 1971 indicated total levels of 325,000 kg SlDDT and 14,000 kg dieldrin in the entire tobacco-growing area. These residues represented 37 percent of the total DDT application to the area since 1961, and were calculated to agree with the half-life disappearance of 3-4 years. Residue analysis of the tobacco crop in 1971 (42 million kg cured leaf) revealed that 52.6 kg yiDDT and 1.97 kg di- eldrin had been removed from the soil. This represents 0.18 percent of the DDT applied in 1971 or 0.03 percent of the :SDDT and dieldrin resident in the soil the same year. Corn silage and hay were calculated to contain 38.3 kg DDT and 0.76 kg dieldrin, hut these residues remained in the water- shed because the crops were used as feed for livestock. Animal products leaving the watershed in 1971 amounted to 54 million kg milk and 6 million kg meat which con- tained only 0.95 kg IDDT and 156 g dieldrin. Total water discharging into Lake Erie from the four water- sheds was calculated to carry 12.6 kg -DDT and 0.86 kg dieldrin in the water and on suspended sediments, represent- ing 0.002 percent and 0.003 percent, respectively, of the -DDT and dieldrin resident in the four watershed soils. 1 Provincial Pesticide Residue Testing Laboratory, Ontario Ministry of Agriculture and Food, Guelph, Ontario, Canada. 2 Pesticide Control Service, Ontario Ministry of the Environment. Simcoe, Ontario, Canada. 3 Fish and Wildlife Research Branch, Ontario Ministry of Natural Resources, Maple, Ontario, Canada. * Sport Fisheries Branch, Ontario Ministry of Natural Resources, Toronto, Ontario, Canada. Fish species caught in Big and Dedrich Creeks contained residues 4 to 15 limes higher than those of species from Long Point Bay and Lake Erie, reflecting the rapid dilution of the creek wtiters within 3 miles of the mouths of these creeks. Lowest pesticide concentrations in fish were about the same as the concentrations in water-suspended sediments. Magnification observed from water to the highest concentrations in large fish was approximately 10'': how- ever, the tissue residue did not exceed the 5 ppm tolerance established in fish. Residues in air appeared to be the result of either spray drift or airborne soil particles rather than volatilization from the soil. Introduction Production of flue-cured tobacco in Ontario is concen- trated in a few counties along the north shore of Lake Erie. Forty-six percent of this cropping area is located in the four watersheds of Big Creek. Big Otter Creek. Dedrich Creek, and Nanticoke Creek, which are cen- tered around Norfolk County (Fig. H. The total area of these four watersheds is 190.340 hectares (ha.) (Table 1). The land is flat or gently undulating and the soil is a deep sandy loam derived from alluvial deposits. Agricultural production occupies 80 percent (152.610 ha.) of the watershed area and tobacco production oc- cupies 10-15 percent of this agricultural land (/). Corn and grain, especially rye. are alternated with tobacco in a 2- to 3-year crop rotation. The dark-sided cutworm (Euxoina messoria) has been a serious pest to tobacco production of the area since 1961. This species has shown a high tolerance to cyclo- diene insecticides (2); thus DDT has been used, in- stead, as an effective early-spring treatment applied to the rye cover crop before plow-down or to soil before planting tobacco (3,4). DDT has also been used to control leaf-eating insects during the season. As a result 184 Pesticides Monitoring Journal Key: Numbers 1-6 ( 9 ) represent sites sampled for DDT and dieldrin residues. FIGURE 1. Four watersheds draining tobacco belt of southern Ontario, Canada Miles and Harris noted a correlation between rainfall and the concentration of insecticides in Big Creek, and reported that this creek carried 50 g DDT per week into Lake Erie in 1970 (6). Although the largest quan- tity of DDT was used in this area, residues in river sedi- ment were no higher than those in recreational areas where only small quantities were used for spraying mosquitoes (7). This was especially evident on the Pre- canibrian Shield where rock was bare or only thinly covered with soil, and DDT readily found its way to lakes and streams following rain. Joint action by the Ontario Ministries of the Environ- ment and of Agriculture and Food restricted DDT to three permit uses in 1970. Included was a use for cut- worm control on tobacco which permitted application of 1.7 kg/ ha, to a rye cover crop at plow-down or a soil treatment before planting. Between 85 and 90 per- cent of the DDT applied under permit in Ontario was used in tobacco production and about 40 percent, 28,970 kg, was used in the study area (Table 1). Most of this study was carried out in 1971 within the boundary of the specified watersheds with the intention of drawing up a balance sheet on the use and dispersal of DDT. Samples were collected from the physical en- vironment, agricultural products, and aquatic biota. of these treatments. Harris and Sans have reported residues between 3 and 4.5 ppm 2DDT in tobacco soils of Ontario (5). Residues increased from 3.1 ppm in 1964 to 4.6 ppm in 1966, and declined slightly in 1969 to 3.4 ppm. The pathway of DDT and its metabolites into the aquatic environment on soil particles is well documented for many parts of the world. Residues found in the water, sediment, and fish in this area have been reported by Miles and Harris, and Frank et al. According to Miles and Harris, highest concentrations were 67 parts per trillion (ppt) in water, 441 parts per billion (ppb) in mud. and 1.0 parts per million (ppm) in fish (6). Frank et al. reported a mean of 91 ppb -DDT in sediments and a residue range of 0.03 to 3.86 ppm in fish depending on the species and its position in the aquatic food chain (7), Field Procedures PHYSICAL ENVIRONMENT Authors selected six sites on the four watersheds drained by Big Creek, Big Otter Creek, Dedrich Creek, and Nanticoke Creek for regular sampling; four of these, 1,4, 5, and 6. were so located that 96, 87, 100, and 81 percent of the tobacco acreage, respectively, were upstream from the site. Sample sites 1. 4. and 5 on Big Otter, Big, and Dedrich Creeks, respectively, were only a short distance up- stream from fiow gauge meters; they were located on sections of each creek where no major inflow of water occurred between any two creeks. Site 6 on Nanticoke Creek was in the tobacco belt; the flow meter was lo- cated downstream at the town of Nanticoke. Water passing the gauge was collected occasionally during the season and compared with water from site 6. TABLE 1. Land acreage, water discharge, and DDT use by watershed, southern Ontario — 1971 Wateruay Watershed Area, ha. Water Discharge (xl0« m^) Agricultural Area, ha. Tobacco, ha. DDT Purchased,' kg Bit! Oiler Creek Big Creek Dedrich Creek Nanticoke Creek TOTAL 79,965 81,815 8,650 19.910 190,340 165.0 143.0 12.9 33.4 354.3 71,430 57,830 6,190 17,160 152,610 6,156 8,562 512 870 16,100 11,175 15,288 927 1,580 28,970 ' DDT purchased by permit. Vol. 8. No. 3, December 1974 185 Creek water samples of 4.5 liters were collected weekly throughout the season from all six sites. Samples were taken from the center of the streams between the sur- face and the water-sediment interphase. Although most waters were clear, all were filtered through Whatman No. 1 filter paper to remove suspended material. Water was quite clear and colorless following this procedure; deposits were too small to analyze. Creek sediment samples weighing 450 g were collected weekly from the same locations by scraping along the creckbed and skimming up surface and mobile sedi- ments. Water and sediments were also collected oflF the mouths of Big and Dedrich Creeks and from Inner Long Point Bay during the spring and late summer. Soils were collected during the spring and fall from one woodlot and several rye and tobacco fields on six to eight farms around each of the six sites. Rye and tobacco soils were sampled from surface to plow-depth. An air monitor was set up in the middle of the tobacco belt close to site 3 on Big Creek; airborne organics were extracted into acetonitrile on 33 days (24-hr periods) between May 13 and August 31. 1971. Between 3.540 and 3.960 liters of air were passed through the acetoni- trile in a 24-hour period. AGRICULTURAL PRODUCTS Flue-cured tobacco was sampled at the end of the 1971 season from 20 farms in the foLir watersheds. Com- posites of 500 g were obtained from each of the five primings by sampling several bales on each farm. Com- posite milk samples of 1.1 liters were obtained from 26 bulk transporters that collected the total volume of milk produced in the four watersheds. Samples were taken directly from the tanker following mechanical mixing. Collections were made over a 2-week period in early summer 1971. Composite beef fat samples of 100 g were collected from abattoirs where animals were known to have come from the study area. A 1-inch cube of fat was removed from the backs of up to 10 carcasses to procure one sample. Samples of corn silage, hay, drinking water, milk, and beef were collected from farms around the six creek sites in October 1971. Silage samples of 25 kg were taken during the normal course of unloading and feed- ing; composite hay samples of 10 kg were obtained from a minimum of 10 bales in storage; water samples of 1 liter were collected from farm ponds and wells; and milk samples of 0.6 liter were taken from holding tanks on farms where silage or hay was obtained. Beef fat was obtained from farms where animals were going for slaughter. AQUATIC BIOTA A total of 289 fish of 24 species were caught either by net or line between 1969 and 1972. Researchers caught eight species predominantly in the creeks and ten in the bay or the lake; six species were common to both creeks and the lake (Table 2). The majority of the fish caught in the creeks were from Big Creek or its tributaries; only a few were obtained from the Big Otter. Those from Dedrich and Nanticoke Creeks were caught close to the mouths and were common to Long Point Bay. All fish were weighed and measured before evisceration. Analyses were conducted on the puree produced by homogenizing the eviscerated fish. Individual fish which were analyzed weighed over 25 g. Analytical Procedures EXTRACTIONS Researchers partitioned 1 liter of filtered water twice by shaking it vigorously for 60 seconds with 50 ml dichloromethane. Extracts of dichloromethane were dried by passing them through anhydrous sodium sulfate and evaporating just to dryness with rotary vacuum at 45° C. Residues were redissolved in 10 ml hexane. Soil and sediment samples were air dried and ground to a fine consistency. A 25-g sample was moistened with 4 ml water and allowed to stand for 12 hours. Soil was ex- tracted with 250 ml 1:1 acetone: hexane (v/v) by shak- ing briskly on a wrist-action shaker for 2 hours. A 100- ml aliquot was removed by filtering and shaking with 300 ml 2 percent aqueous sodium chloride for 60 sec- onds. The hexane extract was dried by passage through anhydrous sodium sulfate; it was evaporated to 5-10 ml by rotary vacuum at 45° C. The acetonitrile solution from the air impinger was reduced to approximately 100 ml by rotary vacuum at 75° C. The solution was partitioned by shaking vigor- ously, first with 100 ml hexane for 60 seconds, then with 500 ml 2 percent aqueous sodium chloride for 30 seconds. The hexane extract was dried by filtration through anhydrous sodium sulfate; it was reduced to 5-10 ml by rotary vacuLmi at 45° C. Hay and silage samples of 25 g dry weight were ex- tracted by blending them with 250 ml 2:1 acetonitrile: water. A 100-ml aliquot was filtered off and partitioned into 100 ml hexane. The extract was concentrated to 5-10 ml by rotary vacuum. Moisture content of the silage and hay was determined by drying at 80° C. Butterfat was separated from milk samples with a de- tergent reagent which had been prepared from 200 g sodium tctraphosphate and 24 ml Triton X-100 in 1 liter distilled water. Butterfat was separated in a water bath at 95°-100° C with 100 ml milk and 100 ml detergent. 186 Pesticides Monitoring Journal TABLE 2. Accumulations of DDT, its metabolites, and dielilrin in several fish species by weight class Species Location No. Average „ Fish Weight, Class, g Analyzed g Weight SDDT DlELDRIN Tissue, ppm Fat, ppm Fish. m8 Tissue, ppm Fat, ppm Fish, Watershed Creeks Watershed Creeks and Lake Erie Cliipeiformes Cyprinidae Carp tCyprinus carpio) Creek Lake All All 1 1 2460 577 0.45 0.05 5.47 3.13 1107 28.9 0.04 0.005 0.49 0.31 98.4 2.89 Percijormes Ceutrtircbidae Rock bass iAmhloplitea rupesttis) Creek Lake All All 5 yd 147 0.76 O.076 20.8 4.15 82.9 11.8 0.04 0.003 LOS 0.16 3.38 0.25 Bluegill tLepomis nuicrochirus) Creek Lake All All 4 IW) 209 0.075 0.021 1.98 4.07 12.2 3.33 0.005 0,001 0.185 0.138 LIS 0.126 Pumpkinsecd (Lepomis gihhosiis) Creek Lake All All 7 5 82 9S 0.60 0.041 13.0 3.15 23.2 4.02 0.024 0.005 0.48 0.37 1.32 0.48 Sihirljormes Iclidiiridae Brown bullhead (Icuduris nebidosiis) Creek Lake All All 1 1(11 137 0.430 0.039 60.0 16.4 86.3 4.17 0.018 0.002 2.68 0.72 3.16 0.23 Amiiformes Amiidae Bowfin (Amia calva) Mouth All 1 765 0.03 2.24 24.5 0.004 0.28 3.06 Crprini formes Calostomidae While bucker {CcUoslomns commersoni) 0-100 -1 6 O.OIO 18.5 0.07 0.004 6.00 0.02 101-200 4 153 0.405 14.9 25.3 0.019 2.19 2.87 over 200 2 472 0.050 51.2 30.4 0.004 1.06 1.89 Cyprinidae Creek chub (Semotilus alromaculatiis) All 5 72 0.678 16.0 33.9 0.036 1.17 1.96 Blacknose dace (Rhinichthys atratidus) All 16 2.2 1.07 13.8 1.50 0.080 1.23 0.12 Spottail shiners (Notropis hmisonius) All 10 5.2 0.382 10.4 3.03 0.018 0.43 0.07 Percijormes Centrarchidae LargemoLith bass {Micropterus salmoides) All 2 153 3.86 51.2 740.0 0.190 4.92 34.9 Salmonijormes Salmonidae Brown trout (Scdmo trittta) All 6 122 1.44 30.7 164 0.047 1.02 5.9 Rainbow trout (Salmo gairdneri) 0-10(10 4 270 1.43 38.6 229 0.050 1.44 7.6 1 00 1-2000 •> 1415 1.08 18.1 1518 0.100 1.68 137 2001-3000 -) 2750 1.03 18.3 2828 0.070 1.24 193 over 3000 1 6400 1.35 25.1 8640 0.070 1.30 448 Vmbridae Central mudminnows {Umbra limi) All 3 3.7 0.81 27.1 3.0 0.050 1.67 0.19 Long Point Bay and Lake Erie Clupeiformes Clupeidae Alewife (Alosa psi'ifdoharengus) All 8 102 0.25 1.13 26.2 ND ND ND Cyprinitormes Catostomidae Northern redhorse (Moxostoma tnacrolepidotttui) All 3 742 0.162 10.8 119 0.009 0.585 6.06 Percijormes Centrarchidae Smallmouth bass (Microplenis dolomieui) 0-1511 9 87 0.12 5.3 10.3 0.005 0.24 0.47 150-300 U 220 0.48 30.8 104.4 O.OOS 0.33 1.21 300-450 8 373 0.54 26.4 194 0.012 0.40 4.66 450-600 6 4X6 1.04 34.1 485 0.020 0.59 9.63 over 600 3 1192 0.88 13.3 1241 0.013 0.25 11.6 Black crappie (Pomoxis nigromacldattts) 0-100 5 75 0.117 7.82 8.71 0.019 1.02 1.12 100-200 2 152 0.174 15.6 26.3 0.006 0.50 0.85 Green sunfish (Lepomis cyanelUis) All 4 199 0.050 2.18 11.0 ND ND ND Percidae Yellow perch (Perca flavescens) 0-50 6 47 0.102 3.33 4.69 0.024 0.87 1.12 51-100 IK 79 0.063 3.16 3.70 0.009 0.54 0.70 101-150 15 126 0.081 7.84 10.13 0.003 0.29 0.36 over 150 1 174 0.043 3.75 7.56 0.008 0.67 1.38 Sciaenidae Freshwater drum (Aplodmotiis grunniens) 0-200 7 171 0.047 0.71 7.90 ND ND ND 201-400 7 316 0.071 0.81 27.0 ND ND ND over 400 6 513 0.245 3.24 128 0.007 0.031 3.45 Serranidae White bass (Morone chrysops) 50-100 9 82 0.101 5.82 7.81 0.004 0.29 0.27 101-150 19 118 0.122 5.70 14.9 0.007 0.44 0.90 151-200 10 180 0.153 3.87 27.8 0.009 0.29 1.69 over 200 4 394 0.470 5.69 229 0.013 0.16 6.52 Salmon ijorm es Osmeridae American smell (Osmerus mordax) 0-20 4 16 0.13 7.4 2.0 0.009 0.52 0.14 20-40 9 28 0.19 8.4 5.3 0.013 0.55 0.34 Salmonidae Coho salmon (Oncorhychns kisittch) 0-1000 7 622 1.14 18.0 829 0.016 0.24 10.2 over 1000 2 28S3 2.38 41.5 6652 0.004 0.07 11.5 Vol. 8, No. 3, December 1974 187 Beef or beef fat containing significant amounts of con- nective tissue was extracted by soxhlet as described for fish tissues. Eviscerated fish were ground into a puree and 10 g were mixed thoroughly with sodium sulfate and Ottawa sand. The mixture was exhaustively extracted by soxhlet over a period of 6 hours with 300 ml hexane. Solvent was removed by rotary-vacuum evaporation and the amount of lipids was determined gravimetrically. COLUMN CLEANUP Water, air, soil, sediment, hay, and silage extracts and up to a maximum of 1 g extractible fat or oil from milk, meat, and fish were quantitatively mixed with 25 g florisil (60/100 mesh) previously deactivated by equi- libration with 5 percent water. A second 25-g portion of similarly deactivated florisil was first added to a chromatographic column, and the florisil. plus extract, was then added to form the upper half of the cleanup column. Columns were eluted with 300 ml 1 :4 dichloro- methane: hexane (v/v) at a rate of approximately 5 ml/min. (8). The eluate was evaporated just to dryness with rotary vacuum at 45° C and redissolved in hexane. Polychlorinated biphenyls (PCB's) were separated from the samples on a charcoal column before gas-liquid chromatographic determinations. PCB results will ap- pear in a later report. GAS CHROMATOGRAPHY Qualitative and quantitative gas chromatographic de- terminations were carried out using the following parameters: Columns: 152-cm-by-3.2-mni-OD Pyrex conlaining 4 percent SE-30 + 6 percent QF-1 on 80/100 Chromosorb W-AW preconditioned 72 hr at 225° C. Detector: Electron capture, either tritium or Ni-63. Temperatures: Injection block 225° C. Column 175° C isothermal. Detectors 200° C (tritium), 275° C (Ni-63). Carrier Gas: Nitrogen at 40 ml/min. Limits of detection for all ingredients except DDT were 0.001 ppb in water, 0.001 ppm in soil, sediment, and plant tissue, and 0.005 ppm in extractible fat of milk, meat, and fish. For DDT the limit of detection was twice these levels. CONFIRMATORY PROCEDURES When concentrations permitted, qualitative DDT con- firmations were carried out by thin-layer chromatog- raphy (TLC) using a 250-^ layer of silica gel developed with 1 percent chloroform in n-heptane and visualized with alkaline silver nitrate. Additional confirmation was achieved by removing the appropriate section of the TLC adsorbent and eluting it with a polar solvent for reexamination by gas chromatography. To form the re- spective dehydrochlorinated derivatives which were measured by gas chromatography (9), p,p'-DDT and p,p'-TDE were confirmed by treatment with alcoholic potassium hydroxide. RECOVERY STUDIES Recoveries of pesticides were checked periodically by direct fortification into the substrate with an acetone solution followed by extraction and cleanup as described above. Fortified milk samples were allowed to stand 24 hours prior to separation of butterfat; soil and sedi- ment samples stood 1 week before extraction. The effi- ciency of the air sampler could not be practically deter- mined. Recoveries from air samples are indicative only of the acetonitrile partitioning and cleanup operations. Averaged percent recoveries were as follow: /^/?-DDE P.[>'-TDE P,p-DDT o.p'-DDJ Dieldrin 98 96 94 94 96 Soil and Milk. Beef, Sediment AND Fish Fat 91 96 89 92 87 91 91 93 86 89 Residues were determined on the basis of dried soil, sediment, silage, hay, tobacco, and the extractible fat of milk, meat, and fish. Moisture and fat contents were used in calculating quantities resident in animal popula- tions of the watersheds. Results were not adjusted to include recovery percentages. Results DDT USE Between 1961 and 1969 about 821,400 kg DDT were used on 183,200 ha. tobacco. Annual acreage varied from a high of 23.850 ha. in 1967 to a low of 13.680 ha. in 1964 (Table 3). Following the DDT restrictions of January 1, 1970, rates per hectare were reduced from about 4.4 to 1.7 kg/ha. and purchase was restricted to permitholders. In 1970 and 1971, 61,330 kg were ap- plied to 33,280 ha. tobacco. Thus, over the 11-year period of 1961 through 1971, a total of 882,730 kg DDT was applied to 216,480 ha. tobacco in the four watersheds (Table 3). SOILS In the four watersheds 40,455 ha. of land is devoted to tobacco production in a 2- to 3-year rotation involving rye, corn, and wheat (Table 4). In 1971 about 16,100 ha. were devoted to tobacco; 24,355 ha. were culti- vated for grain. Rye fields sampled in April and May 1971 had not been cropped with tobacco since 1968 or 1969, but were to be planted late in May 1971. Mean -DDT concentra- tions in soil by watershed varied from 2.33 to 3.44 ppm (Table 4); dieldrin concentrations varied from 0.10 to 0.19 ppm. Authors calculated that a minimum of 173,700 kg DDT and 8,780 kg dieldrin could be resi- dent in these soils. 188 Pesticides Monitoring Journal TABLE 3. Qiiunlilii's of lolnicco i;iown aiui DDT used, souiliern Ontario — 1961-71 YEAR Tobacco Harvested, ha.i DDT APPLIED, kg 1961 22.760 102.100 1962 21,700 97.300 1963 18,530 83,100 1964 13,680 61,300 1965 16.170 72.500 1966 21,880 98,100 1967 23,850 106.900 1968 22.730 in 1.900 1969 21,9(10 98.200 1961-69 (siiblotall 183.200 821,400 1971) 17.180 32,360 1971 16.100 28,970 1970-7i (sublolal) 33,280 61,330 TOTAL 216,480 882,730 ^Agricultural Statistics for Ontario, 1971. (Refer to Literature Cited, reference 1.) TABLE 4. :iDDT and dieldiin in soils of tobacco fields and adjoining woodlots of four watersheds, southern Ontario — 1961-71 Watershed Area. ha. No. Samples Concentration in Soil IDDT. ppm DDE/TDE, % Dieldrin. ppm Residues in Soil, kg IDDT Soil: Rye. Wheat, and Corn Fields ^ Big Oitcr Creek Big Creek Dedrich Creek Nanticoke Creek Total Mean 9.235 12.845 765 1,510 24,355 10 30 10 10 60 2.96 3.44 3.20 2.33 . 2.98 19.7 22.(1 18.0 21. S 20.4 = 0.19 0.14 OKI 0,19 0.16 61,278 99.051 5.487 7,884 173,700 3,934 4.031 171 643 8,779 Soil: Tobacco Fields Big Otter Creek 6.156 12 3.98 19.2 0.19 54.921 2.622 Big Creek 8.562 20 4.36 19.0 (1,14 83.6X3 2.687 Dedrich Creek 512 8 3.95 19.3 11 19 4.522 218 Nanlicoke Creek 87(1 10 3.88 24.8 11,111 7.571 195 Total 16,1(1(1 5(1 150,697 5.722 Mean 4.04 20.6-' 0,16 Total acreage in tobacco 4(1.455 Soil: Woodlots Big Otter Creek 478 3 0.3(17 13.11 Trace 329 4.3 Big Creek 882 5 (1.(148 50,(1 Irace 95 7.9 Dedrich Creek 61 1 0,074 73,(1 Trace 10 0.5 Nanticoke Creek Ull 7 (1.024 33,11 Trace 5.4 0.9 Total 1.522 12 4W,4 13.6 Mean (1,1 n 42.0 = Trace Grand Total 41.977 122 324.840 14,515 ^Residue a( end of rotadon before planting (obacco. ■■'DDE represen(ed 17.3 and 17.5 percent i:DDT and TDE represented 3,1 percent IDDT; hence DDT represented 79,6 and 79,4 perceni IDDT. "Residue in soil after tobacco was harvested. Tobacco fields were sampled in early October 1971, shortly after the crop had been harvested and stalks had been disked under. DDT had been applied in May at plow-down of the rye cover crop or just before planting tobacco. Concentrations of -DDT ranged from 3.88 to 4.36 ppm with a mean of 4.04 ppm (Table 4). Because the same fields were involved in rye and to- bacco, increase in soil residues suggested that at least 2.35 kg/ ha. DDT had been added during the growing season for the two crops. The amount of DDT resident in soil cropped with tobacco in 1971 was calculated at 150.700 kg. Dieldrin residues in soil remained un- changed from rye to tobacco fields and about 5,700 kg was calculated to be present in the tobacco acreage. Composition of the -DDT in soil collected from rye and tobacco fields showed little difference. In both cases almost 80 percent was present as o,p'- and p,p'-DDT and only 20.4 and 20.6 percent was present as the two metabolites, DDE (17.1-17.3 percent) and TDE (3.1 percent ). A total of 1.522 ha. (0.8 percent of the four water- sheds) is devoted to woodlots. Many of these woodlots are adjacent to or part of tobacco farms. Samples of woodlot soils were taken in both spring and fall. Be- catise no woodlot had actually been sprayed, the mean residue of 0.113 ppm (Table 4) represents drift from either aircraft or ground applications, or else the ac- cumulation of deposits from wind erosion. It was cal- culated that 440 kg i:DDT and 14 kg dieldrin were resident in these woodlots. Composition of -DDT in these soils varied greatly from as little as 27 percent to as much as 87 percent o.p'- and p.p'-DUT. Vol. 8, No. 3, December 1974 189 A total ot ,^24.840 kg i:DDT was calculated to be present in the 41,977 ha. of land, including woodlots, involved in tobacco production. This amount repre- sented almost 37 percent of the total estimated applica- tion of 882,730 kg over the 11 -year period. Assuming a half-life of 3 and 4 years for -DDT, the predicted residue falls between 297.000 and 386,000 kg. These calculations SLipport a half-life of 3 to 4 years for DDT in the light sandy loams observed in this study. Total dieldrin residue amounted to 14,515 kg in 41.977 ha. of land; no estimate of use over the 1 1-year period was available. Use of aldrin and dieldrin in the 5 years preceding this study had been slight, largely because of the increasing preponderance of the dark-sided cutworm which exhibited a high tolerance to cyclodiene insecti- cides (2). TOBACCO The largest exported crop which removed organochlo- rine residues from the soil of the four watersheds was tobacco (Tables 4,5). The residue level of i;DDT in cured tobacco leaf varied from farm to farm: mean residues declined from 2.52 ppm in sand leaves to 0.66 ppm in tips. The major portion of this -DDT was o,p'- and p,//-DDT; 21-30 percent was DDE, and 4-9.5 per- cent was TDE. The anunmt of -DDT in the 1971 crop was calculated at 52.6 kg. If it were all derived from the 1971 application, this would represent 0.18 percent of the 28,970 kg applied. If it were derived from -DDT resident in tobacco soils during the growing season, it would represent only 0.035 percent. Dieldrin residues were insignificant, declining from 0.09 ppm in the sand leaves to 0.02 ppm in the tips. The amount of dieldrin calculated to be present in the crop was 1.97 kg, representing 0.035 percent of that resi- dent in the planted tobacco acreage. SILAGE .Silage in the four watersheds is produced largely from corn which is often inserted in the tobacco-grain rota- tion in place of wheat for feeding dairy or beef cattle. In the four watersheds approximately 6,028 ha. corn produced 172.7 x 10'' kg silage during 1971 (Table 6). The weighted mean of this silage crop contained 0.339 ppm -DDT and 0.005 ppm dieldrin on a dry-weight basis. Total quantity of -DDT in the whole crop was calculated at 34.2 kg and 536 g dieldrin. Composition of DDT in the silage varied greatly from a high of 74 percent to a low of only 21 percent o,p'- and p.p'-DDT. Likewise, the />,/>'-DDE content ranged from 22 to 77 percent; TDE content ranged from 2 to 6 percent. These findings suggest that residues resulted from spray drift where o.p'- and p,p'-DDT were high, and from soil uptake or soil contamination where DDE was high. TABLE 5. 1.DDT and tlicUlrin in ciintl tohacco Icaj of fai/r \nilci.\/uu/\. soiillui n Oiitciiiu — 197 1 Content in Dried CitRi d Priming C URtl> LEAf^ ■ [*R01)L)(. HON ( • null) kg) Leaf ppm DDT Mliaboliils, r; Residues in Crop, ky (Lr-AVESi IDDT Dieldrin DDE TDE IDDT Dieldrin 1 Sand 4,211 2.52 0 09 21 4 9 5 10,61 0.18 2 .Seconds 5,894 2.08 0 07 21 1 7,7 12,26 0,41 .1 Thirds 8,0011 lis 0 1)7 24.6 5,9 9,44 0.56 4 Fourths 1.1.89.1 0,98 0 01 29,6 4,1 11,62 0.42 5 Tips 111.104 0,66 0 02 28,8 4.5 6,67 0,20 Total 42,102 52.60 1.97 Mean 1.25 0.05 26.7 5.6 'Records of Ontario Flue-Cured Tobacco Marketing Board, 1971. (Refer to Literature Cited, reference 15.) TABLE 6. :::DDT in corn siUiffc ami 1 ay of four nalfr.slu'tis, soiilhcrn Ontario- -1971 Crop Acreai.e. ha Croi' Prodl'i, HON ( ~. 1000 kgi MolslURI . % Residues in Dried Feid Residues in Total Fied Watershed IDDT. ppm Dll 1 DRIN, ppm IDDT, Dieldrin, g Corn Selac.f, Bit; Otter Creek -1,918 114,020 42,0 0,141 0 004 22,68 265 Big Creek 1,772 49,990 40,5 0,151 (I DOS 10.50 2.18 Dedrich Creek 61 1.670 IS'^ 0 260 0.004 0.27 4 Nanticoke Creek 257 7,010 41 s 0 18S 0.007 0,26 29 Total 6,028 172,710 14,21 516 Mean 0,119' 0,005 Total Mean 11,792 61,600 4.10 222 ' ^DDT residue consisted of 21-74 percent DDT. 22-77 percent DDE. and 2-6 percent TDE. -Hay samples were scarcer than silage samples; hence individual watersheds are nol listed. ' i:DDT residue on hay by watershed ranged from 0.05 to 0.09 ppm present mainly as actual DDTI75 percent). 190 Pesticides Monitoring Journal TABLE 7. ZDDT and Jieldrin in milk and beef of four watersheds, southern Ontario — 1971 Watershed Annual Producik: (X 1000 kg) iDDT IN EXIRAt TIBLi: Fat,' ppm Composition of DDE AND TDE, % DiELDRIN IN EXTRACIIBLE Fat, ppm Residues in Annual Supply, g IDDT Dieldrin Milk Production '■ Big Oner Creek 32.060 0.183 79.1 0.031 234,7 39.8 Big Creek 9,716 0.234 82.9 0.050 90.9 19.4 Dcdrich Creek 70 0.2311 80.1 0.045 0.6 0.1 Nanticoke Creek 11,716 ().25« 86.0 0.057 120.9 26.7 Total 53,562 447.1 86.0 Mean 0.188 81.0 0.035 Beef Production -i Big Otter Creek 4,182 0.340 82.4 0042 355.5 43.9 Big Creek 1,468 0.325 81.5 (1,051 119.3 18.7 Dedrich Creek 57 0.298 80.9 0.030 4.2 0.4 Nanticoke Creek 275 0.267 85.0 0.052 18.4 3.6 Total 5,982 497.4 66.6 Mean 0.333 82.3 0.045 'Average butterfat content: 4%; average beef content: 25%. = Reference Records, Ontario Milk Marketing Board, Toronto. (Refer to Literature Cited, reference 16.) ^Annual Livestock Market Review, 1971, Canada Department of Agriculture. (Refer to Literature Cited, reference 17.) HAY Hay samples of limited quantity were obtained from fields in the four watersheds but not from tobacco farms. Residues of i:DDT varied from 0.05 to 0.09 ppm with a mean composition of 75 percent actual DDT. This suggested that the deposition of DDT was recent, possibly resulting from spray drift. Total hay production in the four watersheds was 61.6 x 10" kg in 1971; this contained 4.1 kg 2DDT and 0.22 kg dieldrin (Table 6). MILK Milk produced in the four watersheds totalled 53.6 x 10" kg a year (Table 7). The weighted mean residue of -DDT in the butterfat from the four watersheds was 0.188 ppm. Where dairy and tobacco production oc- curred close together, residues ranged from 0.183 to 0.258 ppm. On three farms that raised tobacco and maintained sizable dairy herds. 2DDT residues in butterfat were 0.39 ppm; average butterfat in the milk was 4.1 percent. These residues were considerably higher than the provincial average of 0.134 ppm re- ported by Frank et al. (10). Composition of -DDT in the general milk supply ranged from 79 to 86 percent DDE and TDE. All milk except that from Big Otter Creek watershed had residues of dieldrin above the 1967-69 provincial average of 0.031 ppm (10). A total of 447 g i:DDT and 86 g dieldrin were present in the annual milk sup- ply of the four watersheds (Table 7). The 447 g DDT present in milk represented only 1.2 percent of the DDT found in hay (4.1 kg) and corn silage (34.1 kg); the 86 g dieldrin in the milk represented 1 1 percent of that in hay (222 g) and corn silage (536 g). BEEF Almost 6 million kg meat a year are produced in the four watersheds (Table 7). Concentrations of -DDT ranged from 0.267 to 0.340 ppm with a weighted mean of 0.333 ppm. Over 80 percent of the i:DDT was present as the two metabolites DDE and TDE. Beef contained a mean of 25 percent fat; the calculated amount of i:DDT in the meat supply was about 500 g. a quantity slightly greater than that found in the annual milk supply, but representing only 1.3 percent of that present in the corn silage (34.1 kg) and hay (4.1 kg) produced in the area. Dieldrin residues in beef fat ranged from 0.030 to 0.052 ppm with a weighted mean of 0.045 ppm. Total dieldrin present in the annual production of beef was less than 70 g, slightly lower than the amount found in the milk supply; this represented about 9.3 percent of the di- eldrin present in corn silage (536 g) and hay (222 g) (Table 7). LIVESTOCK WATER SUPPLIES Water from shallow wells and from spring- and surface- fed farm ponds was collected to determine contamina- tion of livestock water supplies. Of 14 wells, 3 had waters containing residues of 4, 40, and 50 ppt 2DDT, respectively, but none contained dieldrin. In 1 1 wells neither -DDT nor dieldrin could be detected. Six spring-fed and nine surface-fed farm ponds were sampled (Table 8); 2DDT was detected in two spring- fed ponds at 1 and 50 ppt, and in four surface-fed ponds at 9, 35, 60, and 80 ppt, Dieldrin was found in only two surface-fed ponds at 5 and 8 ppt. These data give mean residues of 6.7 ppt -DDT in 14 wells, 8.5 ppt i:DDT in 6 spring-fed ponds, and 20 ppt -DDT and 1 .4 ppt dieldrin in 9 surface-fed ponds. AIR On 35 days (24-hr periods) between May 1 and Au- gust 31, 1971, air was sampled for DDT. Residues oc- curred on 19 of the 35 days (Table 9). Highest mean concentrations occurred in May (74 ng/m^); lowest Vol. 8, No. 3, December 1974 191 concentrations occurred in June (2.3 ng/m^). On 10 of the 19 days only o,p'- and p,p'-DDT were detected; on 2 other days when o,p'- and p,p'-DDT were high. TDE was also detected. When found, TDE represented 4.3 percent of the -DDT. a normal percent component in the formulated product. Hence these residues were con- sidered to have been the result of spray drift. Spraying operations appeared to have occurred on 12 days: 2 in May, 5 in June. 4 in July, and 1 in August. Mean con- centration in air on these spray-days was 46.7 ng/m'' air. On 9 of the 12 days, the concentration in air was less than 10 ng/m'', but on the other 3 days i;DDT concentrations were 15, 48, and 469 ng/m-'. For 7 days, DDE and TDE were present at 21 percent and 14 percent, respectively, of the i;DDT (Table 9). The presence of these metabolites suggested that resi- dues were derived from either airborne dust particles or volatiles. The fact that the ratio of DDE + TDE/ DDT was 1:2 and fairly constant for the 7 days sug- gested that these residues were derived from airborne particles rather than from volatiles. Mean concentration on these 7 days was 34.5 ng/m-'' liDDT. Mean residue in air for the 35 days was 27.4 ng/m.-' No dieldrin was detected on any sampling days. Snow samples were collected in late winter from three main areas in the Big Creek watershed in order to measure DDT fallout. Samples were divided into upper and lower levels to represent early-winter and late- winter snowfalls. Water from the early snowfalls con- tained an average of 43 ppt -DDT with a high of 205 ppt. Late-winter snow contained only 4 ppt (Table 10). Dieldrin was also highest in early-winter snow with a mean of 10 ppt as opposed to 0.7 ppt in late- winter snow. Composition of the -DDT suggested that these residues were derived from airborne particles and not volatiles. It is assumed that residues detected in air and snow might explain the movement of DDT, its metabolites, and dieldrin from the tobacco crop to other crops such as hay and silage that are not sprayed and often are not grown on soil in which tobacco has been produced. CREEK AND BAY WATER Big Otter, Big, Dedrich, and Nanticoke Creeks dis- charged 165, 143, 12.9, and 33.4 million nr' water,, respectively, into Lake Erie during 1971 (//). Monitor-- ing of waters for organochlorine insecticides was ex- • tended on Big Creek from February to October, a pe- riod that accounted for 81 percent of the discharge. On the other three creeks, monitoring extended from March ' to October and accounted for 60 to 69 percent of the discharge (Table 11). TABLE 8. Residues of ZDDT and dieldrin in surface-fed and spring-fed ponds and farm wells, southern Ontario — 797/ Source of Water No. Sources Sampled No. Residues Detected SDDT Dieldrin Residues Detected, ppt i:DDT Dieldrin Surface-fed farm ponds Spring-fed farm ponds Farm wells 9 6 14 5 4 11 7 6 4 9. 35, 60, 80 1,50 4, 40, 50 5,8 ND ND NOTE: ND = not detectable. TABLE 9. DDT and its melaholites in air from Big Creek watershed, soulhern Ontario — 1971 Month of No. Days in No. Days Residues 2DDT DDT Composition. % Sampling Operation Detected Mean Residue in Air, ng/m = DDE TDE DDT May 9 4 73.9 5.2 6.0 88.8 June 12 4 2.32 0 0 100 July 5 5 23.8 4.0 2.0 94 0 August 9 6 8.11 9.4 13.6 77.0 May-August 35 19 27.4 5 2 6 1 88 7 Suspected sprayings 12 46.7 0 4.3 93 7 Suspected soil or dust 7 34.5 20.8 14.2 65.0 TABLE 10. ^DDT and dieldrin in snow from Big Creek watershed, southern Ontario — 1971 Snow La-yer No. No. Days Residues Samples Detected Mean Residue in Water, ppt > IDDT Dieldrin SDDT Composition, % DDE TDE DDT Upper level Lower level 6 6 4 4 4 43 0.7 10.0 12 14 0 10 68 76 192 Pesticides Monitoring Journal TABLE 1 1. Concentration and amount of IDDT and dieldrin in creeks, soiitlwrn Ontario — 1^71 Waterflow ' Concentrations IN Water, ppt - Composition, % Residues, g Month (X lO-im'') IDDT Dieldrin DDE TDE DDT i:DDT Dieldrin Big Otter Creek March 28,659 15.2 1.8 18 48 34 436.0 51.6 April .11,406 ISO 3.0 22 33 45 565,0 94,2 May 14,240 21,4 0.4 13 35 52 305.0 5.70 June 7,214 14.6 0.9 46 14 40 105.0 6.49 July 8,169 9.7 0.8 32 48 411 79.2 6.54 August 4,910 5.7 0.1 32 28 40 28.0 0.49 September 5,486 4.8 0.0 30 50 2(1 26.3 0.00 October 7,901 4 7 0,1 19 60 21 37.1 0.79 Mar.-Oct. 107,985 14.6 1,5 1,581.6 165.31 % Total 65.4 76.9 87.9 Jan., Feb., Nov., Dec. 57,118 4.7 0.1 268.0 5.71 Annual total 165, IW 1,849.6 171.52 Annual mean 11.2 1,0 Big Creek February 13,628 5.5 0.2 17 ' 61 21 75.0 2.73 March .17,4(18 30.0 0.8 IS 5(1 32 1,122.0 29.9 April 24,062 43,1) 3.0 15 5(1 35 1,035.0 72.2 May 11,383 16. K 1.0 17 26 57 191.0 11.4 June 7,776 11.8 0.2 51 17 32 91.8 1.56 July 4,910 8,8 1.5 311 28 42 43.2 7.37 August 4,910 6,9 (1,1 26 29 45 33.9 0.49 September 5,875 11,0 (1,1 16 29 55 64.6 0.59 October 6,250 7.1 0.1 17 45 38 44.4 0,63 Feb. -Oct. 116,202 23.2 1.1 2,700.9 126.87 % Total 81.1 94.0 96.9 Jan., Nov., Dec. 27,100 6.3 0.1 171.0 2.71 Annual total 143,302 2,871.9 129.58 Annual mean 20.0 0.9 DEDRtCH Creek March 554 23.1 2.5 15 58 27 12.3 1.39 April 3,339 20.0 3.0 15 5(1 35 66.8 10.0 May 1.629 18,1 0.4 14 41 45 29.5 0.65 June 890 45.1 0.4 9 46 45 40,1 0.36 July 281 8.1 0.8 28 36 36 2.28 0.22 August 250 8.1 0.4 28 36 36 2.03 0.10 September 337 7.4 0.3 27 41 32 2.49 0.10 October 379 6.5 0.2 25 47 28 2.46 0.03 Mar.-Oct. 7,659 20.7 1.7 158.46 12.90 % Total 59.5 82.4 92.5 Jan., Feb., Nov.. Dec. 5,215 6.5 0.2 33.9 1.04 Annual total 12,874 192.36 13,94 Annual mean 14.9 1.1 NiNTicoKE Creek March 12,812 11.1 1,2 15 57 28 142.2 15,37 April 4.882 16.(1 2,0 25 48 37 78.1 9.76 May 1.821 23,9 1,3 18 31 51 43.5 2,37 June 2,497 IS 9 2.5 34 15 51 47.2 6,24 July 241 11,2 1.5 25 35 40 2.70 0.36 August 76 5,5 0.1 25 35 40 0.42 0.01 September 242 7 0 1.0 14 29 57 1.69 0.24 October 455 6,2 0.5 15 48 37 2.32 0.23 Mar.-Oct. 23,025 1,1,8 1.5 318.63 34,53 % Total 68.9 77.9 80,6 Jan., Feb., Nov., Dec. 10,371 6,2 0.5 64.3 5,19 Annual total 33,397 382.93 39,77 Annual mean 11.5 1.2 Four Watersheds Annual total 354,676 5,296.8 354.81 Annual mean 14.8 1.0 Inner Long Point Bay Mouths of Big and Dedrich Creeks \2 s (13 45 ST 33 Middle of bay (i sy.ui6 ■ 409.9 8.91 miles) 1..1 ND 100 0 11 Edge of Inner and Outer Long Point Bay Trace 1 ND 1 -' — 1 Water Flow Data. Environment Canada. (Refer to Literature Cited, reference 11.) -Ppt = parts per trillion. -Bersi and McCrimmon. (Refer to Literature Cited, reference 1-.) Vol. 8, No. 3, December 1974 193 The highest concentration of -DDT in the waters of these four creeks occurred during the spring months, March to May. when discharge volumes were highest. Concentrations rose to 21. 43. 45. and 24 ppt. respec- tively, in Big Otter, Big, Dedrich. and Nanticoke Creeks coincident with spring thaw and runoff, cultivation of land, and spraying of the rye cover crop for cutworm control. Concentrations in water declined in midsummer to 5-8 ppt in spite of rainfall that was heavier in the period from July to September (Table 12). 2DDT in water in Long Point Bay just off the creek mouths con- tained lower concentrations (12.5 ppt) than either the waters from Big Creek (20 ppt) or Dedrich Creek (14.9 ppt). indicating mixing and dilution as water enters the bay. Three miles off the mouths of these creeks in the middle of the Inner Bay. residues had declined to 1.3 ppt iDDT and were all present as DDE (Table 1 1). Water collected at the juncture of Inner and Outer Bays con- tained only traces of DDT. It is estimated that Inner Bay contained 89.106 thousand m'' water (12) which could contain 410 g -DDT and 8.9 g dieldrin. During the months of February. March, and April, the largest fraction of -DDT was present as TDE (Table 11, Fig. 2). The parent compound DDT was lowest in February and increased to become greater than TDE in May; the DDE fraction remained small and unchanged. In the period of May to September. DDT comprised the greatest fraction as it rose to a peak; it then declined in October. The TDE fraction declined to its lowest level in June, but increased again in July. The DDE fraction almost doubled during June to August to 30 percent. By October TDE again became the predom- inant fraction; DDE and DDT declined and remained in roughly equal proportions. Increase in DDT coin- cided with its use on rye and the cultivation of the soil for planting the tobacco crop. The largest resident load of -DDT discharged by the four creeks occurred between March and April when the water discharge volumes were at their highest; this was over 1000 g -DDT in the case of Big Creek dur- ing March and April (Fig. 3). Big Otter Creek, which discharged a greater volume of water than Big Creek, delivered only half the quantity of DDT to Lake Erie during these peak months. Dedrich. a small creek, dis- charged only 67 g in April. Site 6. the main sampling site on Nanticoke Creek, was located in the tobacco belt; the flow meter was down- stream. Water at site 6. which was sampled regularly, contained a peak of 24 ppt in May and a low of 5.5 ppt in August. The six water samples collected at the flow meter near the town of Nanticoke had a peak of 35 ppt in May and 4 ppt in August. It was not clear where the additional DDT between the two sites origi- nated because the volume of water almost trebled. Be- cause the residue data at site 6 were more complete than at the flow meter, these site data were used to estimate residue loads. Nanticoke Creek appeared to carry its largest load of DDT. 142 g. in March. SO- 30- ^ 20- 3. "^- SED ^ EN J. ' -h r^ r^ J, .1 2 — a, A- : ■ : D T D HE E "Ti -L IJL. 0 " 50- 0 •2 40- 10- 1 > , n h E n i 1 1 n ' 7_ ^ 2 3 ■r ! pf r^ r" 3 r J. ) f£B MAC APR ^AA■< JUNt JULY AUG StPT oc? 1 FIGURE 2. Monthly composition of IDDT in water and sediment of creeks in study area, 197 1 TABLE 12. Frequency and amoiDit of rainfall. 197 1 '^ Frequency of Rainfall, Days Range, cm Month O.OI-l.OO 1.01-2.00 2.01-3.00 Over 3.01 Rainfall, cm = January 10 0 0 0 4.72 February in 1 0 0 8.26 March 3 2 0 0 3.68 April 5 2 0 0 3.73 May 8 0 0 1 4.06 June 8 2 0 0 4.27 July 6 1 1 0 5.36 August 6 0 t 1 9.68 September 10 0 1 0 4.55 October 6 1 0 0 3,86 November 9 2 0 0 5.00 December 9 5 0 0 11.63 Total 90 18 4 2 68.80 iClimatological Station Report, Canada Department of Agriculture, Delhi, Ontario. 1971. (Refer to Literature Cited, reference 18.) = Of total rainfall, snow comprised 3.05 cm in January. 3.56 cm in February. 2.03 cm in March. 1.27 cm in April. 1.14 cm in November, and 2.06 cm in December. 194 Pesticides Monitoring Journal 1250-1 1000 — E 750- < X 500- Q < o 250- 2 3 4 FEB MAR 1 BIG CREEK 2 — BIG OTTER CREEK 3 NANTICOKE CREEK 4 DEDRICK CREEK APR 1 2 JUNE 3 4 JULY SEPT rn^ OCT 197 1 FIGURE 3. '^DDT residues in discharge water entering Lake Erie from the four creeks in the study area, 1971 (NOTE: no data available on creeks 2, 3, or 4 for February) Because the concentration of -DDT in creek water was highest in the spring and lowest in the fall, an estimate based on lower concentrations was made for quantities of DDT and dieldrin through late fall and early winter. Big Creek carried the greatest annual residue load of i;DDT which was estimated at 2,872 g. Big Otter Creek carried the second-highest load, 1,850 g, and Nanticoke and Dedrich Creeks carried 383 and 192 g, respectively. Collectively, all four watersheds carried a total of 5,297 g/year, an average of 14.5 g/day, into Lake Erie. This represented that portion of DDT and its metabolites that was either dissolved in the water or on particles that passed through Whatman No. 1 filter paper and not that transported on removable suspended sediment. Concentrations of dieldrin in creek water were also highest in March and April, rising to a level of 2 to 3 ppt. Concentrations declined during the summer and fall to between one -fifth and one-tenth these levels. Dieldrin concentrations represented between one-tenth and one-hundredth the level of -DDT. Residues of dieldrin entering Lake Erie were also highest in March and April; the greatest quantity, 94 g, was carried in the peak month of April by Big Otter Creek. The largest qLiantity carried by Big Creek, 72 g, occurred in April. Nanticoke and Dedrich Creeks carried peak amounts of 15 and 10 g, respectively, in these same months. The annual quantity reaching Lake Erie was 172 g from Big Otter Creek. 1 30 g from Big Creek, 40 g from Nanti- coke Creek, and 14 g from Dedrich Creek. Hence the annual discharge of the four watersheds was 354.7 g, almost 1 g/day. CREEK AND BAY SEDIMENTS Sediments taken at the interface between creekbed and water showed weekly variations in the concentrations of -DDT and dieldrin as they shifted downstream (Table 13). Highest mean -DDT residues were observed in sediments from Dedrich Creek (141 ppb) but were much lower in Nanticoke Creek (55.6 ppb). Big Otter Creek (41.3 ppb). and the lower reaches of Big Creek (38.7 ppb). Sediments in Dedrich Creek contained almost 200 ppb -DDT in June; however, this level de- clined steadily to 93 ppb by September. Greatest resi- due levels in Big Otter Creek, 58.6-61.0 ppb, were found in August and September. The highest sediment Vol. 8, No. 3, December 1974 195 residues in NanticoK=' Creek, 70-95 ppb, were measured between April and lune at site 6; these levels declined steadily to 23 ppb by September. At the flow station downstream, sediment residues ranged from 25 to 1 ppt with a mean ol 10 ppb, less than one-fifth the residue at site 6. In Big Creek, highest sediment residues of -DDT, 45-51 ppb, were observed on the lower reaches between June and August. In the middle reaches, sediment resi- dues from April to September were relatively imifomi (13-21 ppb); in the upper reaches the sediment residue level reached its peak, 42 ppb, in May and declined until August. Sediment collected from Long Point Bay off the mouths of Big and Dedrich Creeks showed -DDT residues of 28.7 ppb: they were only 4.3 ppb 3 miles into the bay (Table 13). Peak concentrations in sediment occurred several months after peak concentrations in water and com- position of -DDT in sediment ditfered from that ob- served in water (Fig. 2). The parent compound DDT predominated in sediments from Big Creek and was generally greater than the DDE fraction in the other three creeks. In Big Creek. DDE was also generally higher than TDE. In early spring and late summer TDE reached its highest percentage, but between these times, in May and June, it declined to its lowest level (Table 13). Composition of -DDT ui sediment from Nanticoke Creek differed from that of Big Creek. In the Nanti- coke, TDE predominated in samples collected during 4 of the 6 months; DDT and DDE were in approxi- mately equal qiiantities. Sediments from Dedrich Creek contained predominately DDT during April, May, and June, and predominately TDE during July, August, and September. In Big Otter Creek the composition of -DDT showed no definite pattern (Table 13). In Long Point Bay, TDE predominated in samples col- lected near the mouths of Dedrich and Big Creeks, but 3 miles out in the bay, TDE and DDE were present in about equal amounts. Dieldrin residues in sediments from all four creeks were of a similar level; the highest mean residue, 2.1 ppb, was found in Big Otter Creek, and the lowest, 0.7 ppb, was from Dedrich Creek. Residues in sediment in Long Point Bay close to the OLitlets of Big and Dedrich Creeks contained 0.6 ppb dieldrin although dieldrin could not be detected 3 miles out in the bay. Dieldrin residues in general appeared in quantities between one- twentieth and one-fortieth the levels of -DDT. In 1971 Environment Canada (II) reported that Big Otter Creek carried 54,368,000 kg and Big Creek carried 13,438,000 kg suspended sediments into Lake Erie. Based on the residues found in sediments in this study, movement into Lake Erie from these two creeks could amount to 1.60 kg i:DDT and 0.114 kg dieldrin (Table 14); movement from Big Creek could have been 0.39 kg i:DDT and 0.013 kg dieldrin. These amounts represent a similar qtiantity of insecticide car- ried by filtered water in Big Otter but only one-seventh that carried by the filtered water of Big Creek (Tables 1 1.14). FISH A total of 289 individual fish belonging to 24 species were caught for analysis from the creeks, the lake, or the bay. Of these 24 species, eight were caught only in the creeks, ten were caught only in Long Point Bay or in Lake Erie off the mouths of the four creeks, and six were common to the two systems (Table 2). All species caught in the lake, except coho salmon (Oncoihychiis kisutch), had mean tissue residues below 1.0 ppm -DDT. Of the eight species caught in the creeks, four had mean tissue residues over 1 ppm; these were brown and rainbow trout (Salmo trutta and S. gairdncri). largemouth bass (Aficroplenis salmoUles). and blacknose dace (Rhinichthys atratulus) (Table 2, Fig. 4). The highest tissue residues, 3.86 ppm, were found in largemouth bass. Of the eight species caught in the creeks, five had mean extractible fat residues over 15 ppm -DDT. These included largemouth bass with the highest level, 51 ppm; brown and rainbow trout; creek chub (Semotiliis alromaciilatus): and central mudminnow (Uiuhra limi). Residue levels over 1 5 ppm -DDT in the fat were found in two species caught in the lake; these were smallmouth bass (Micioptenis dolomieiii) and coho salmon. Although the species were not common to the two bodies of water, there was a greater concentration of -DDT in these species confined to the creeks. These differences were very apparent in species caught in both systems. Of six species common to the two systems, tissue residues of -DDT ranged from 4 to 15 times higher in those members caught in the creeks than in those caught in the lake (Fig. 4). Dieldrin residues were over 0.1 ppm in the tissues of only one species, largemouth bass. Only one species, blacknose dace, had mean dieldrin residues between 0.05 and 0.1 ppm. Both species were caught in the creeks. Of the 10 species caught in the lake, three had undetectable dieldrin residues and many of the re- mainder had residues at or below 0.01 ppm. When resi- dues in extractible fat are considered, only one species caught in the lake, black crappie (Pomoxis nigro- maciihitus). had dieldrin residues over 1 ppm. However, seven species caught in the creeks contained residues over 1 ppm and in largemouth bass the residues were almost 5 ppm. 196 Pesticides Monitoring Journal TABLE 13. Composition and concentration of ZDDT and dicldrin in sediments of four water courses in four watersheds, southern Ontario — 1971 Month Dried Sediment, ppb Composition, 2DDT DiELDRIN TDE DDT Big Creek— Upper Reaches; Sand April 19.4 ND 26 36 38 May 41.8 0.8 30 17 53 June 17.6 0.6 39 24 37 July 15.5 0.4 54 16 30 August 10.9 0.6 41 26 33 September 11.6 0.4 29 36 35 Mean 19.5 0.5 Big Creek — Middle Reaches; SandV Silt April 20.8 2.0 43 31 26 May 19.0 0.7 36 24 40 June 12.8 0.2 33 20 41 July 19.7 11.4 29 26 45 August 15.7 0.4 22 21 57 September 20.0 U.5 20 26 54 Mean 18.0 0.7 Big Creek — Lower Reaches; Fine Sand April 25.4 0.4 20 25 55 May 30.6 2.0 29 16 55 June 51.4 0.5 34 13 53 July 50.2 2.1 33 26 41 August 44.8 0.4 29 34 37 September 29.7 0.4 28 34 38 Mean 38.7 1.0 Big Otter Creek; Sandy Silt April 24.4 3.0 33 43 24 May 34.6 0.3 38 23 29 June 40.5 1.4 53 21 26 July 28.8 1.3 29 36 35 August 58.6 3.6 27 28 45 September 61.0 3.0 25 36 44 Mean 41.3 2.1 Dedrich Creek; Sandy Silt April 162.4 0.1 22 25 53 May 129.6 1.5 24 36 40 June 198.2 0.8 19 38 43 July 138.3 1.3 2'> 48 30 August 124.4 0.1 20 43 37 September 93.0 0.4 20 45 35 Mean 141.0 0.7 Nanticoke Creek; Sandy Silt April 73.0 2.4 45 25 30 May 94.8 0 30 43 27 June 70.3 1.4 23 34 43 July 45.7 2.4 32 46 32 August 26,6 1.4 35 43 TT September 23.0 3.0 30 39 31 Mean 55.6 1.9 Long Point Bay Mouth: May-September Middle of bay: 3 miles 28.7 4.3 0.6 ND 18 46 72 48 10 6 NOTE: ND = not detected. TABLE 14. IDDT and dieldrin carried on suspended solids by Big Creek and Big Otter Creek to Lake Erie — 1971 Suspended Solids, kg/yr Average Concentration, ppb Amount Carried to Lake Erie, kg Creek iODT Dieldrin SDDT Dieldrin Big Otter Creek Big Creek Total 54,368,000 13,438,000 67,806,000 41.3 38.7 2.1 1.0 1.60 0.393 1.993 0.114 0.013 0.127 Vol. 8, No. 3, December 1974 197 C. Salmon S.M. Bass Alewtfe A. Smelt N. Redhorse W. Bass B. Crappie F. Drum Y. Perch G. Sunfish Bluegill 1 Carp 1 B. Bullhead • Pumpkinseed _ _r" R. Bass--' W. Sucker S. Shiner C. Chub C. Mudminnow B. Dace R. Trout B. Trout L.M. Bass BAY AMD LAKE FISH CREEK FISH I 2 TOTAL DDT, ppm FIGURF 4. -DDT ii>mcnlrali<>i\s in lissiicM of 23 fish species caHi;lit in creek and liike cn\ ironnienls, soiillieni Ontario — 1971 Creek Fish Among the species caught in the creeks, hiacknose dace and spottail shiners (Nolropis hiidsonius) were members oi the lowest trophic levels. Yet their tissue and fat residues of both -DDT and dieldnn were com- paratively high. Boltom-leeding white suckers (Caiosto- iiiiis comincrsoiu). which have lower tat content in the tissues, had lower residues of DDT and dieldrin than those in dace and shiner; but based on exiractible fat their residues were higher. A marked increase in -DDT was observed in both extractibic fat and total body bur- dens as the average weight of white suckers increased. Creek chub, an important link in the aquatic food chain, contained almost 0.7 ppm -DDT in the tissue and 16 ppm in the extractible fat. Dieldrin levels were 0.04 ppm in the tissue and 1.17 ppm in the fat. Piscivores caught in the creek included largcmoLith bass, and brown and rainbow trout. Largemouth bass ac- cumulated the highest residues of -DDT and dieldrin ill tissue and extractible fat. As the size of the rainbow trout increased, tissue and fat concentrations showed little change although the body load increased rapidly, [n fish weighing an average of 6.400 g body load. 8.6 mg -DDT and 0.4.'^ mg dieldrin were accumulated. Body burdens in the creek-caught species increased from 0.07 iig -DDT in tiny white suckers weighing an average of 6 g to S.640 iig in the largest rainbow trout. This represented an accumulation of 1.2 x 10^. With dieldrin. the obscr\ed increase was from 0.02 ng in tiny white suckers (Table 2) to 448 ug in large rain- how trout, an accumulation o\ 6.4 x 10-' (Table \5). Lake f'isli Plankton leeders caught in the lake had -DDT resi- dues up to 0.2."^ ppm in the tissLie biit only trace quanti- ties of dieldrin. The bottom-feeding redhorse (Moxo- stiiinii i>icicn>lci>i(liyliiiii) contained low residues similar lo those oi the plankton leeders. Black crappies feed- ing on crustaceans and small fish showed only slightly ele\ated residues, .\moiig the piscivores. smallmouth bass contained an elevated tissue and fat concentration of IDDI and dieldrin and exhibited a buildup in body loads with increasing tish size. Coho salmon contained the highest residue of i:DDT. The range Irom the lowest hodv load of 2 ug -DDT in American smelt iOsnienis niorclax) to the highest body load ol 6.652 ug in coho salmon represents an accumulation of 3.,^ x IIF. With dieldrin the range was from a nondetectable level in green sunfish (Leponiis cyiinelliis) to 11.6 iit; in smallmouth bass. 198 Pesticides Monitoring Journal TABLE 15. Biomagnificalion of ^DDT aiul dieldrin in the aquatic ciiviroiiniriit of watcrshcil creeks ami Lake Erie, soiillwrn Ontario — 197 1 iNscciEcinn Residues IN 1 g Magnification Watershed Creeks ilDDT Waler 1.5 X 10 " Scdimenl 4.1 y 10" 2.7 X 10-' While sucker (smallest ) 1.2 .■ 10 ' 8.0 X 10- Rainbow Iroiit (larjiesl) 1.4 •- 10 'i 9.3 X 10' Largemoiuh bass (liiiihesi 4.« ,• 10 « 3.2 X lO'' residues) Dickirin Waler 1.0 X 10-1- Sediment 1.9 X 10-^' 1.9 X W While sucker (smallest) 3..1 X 10-" 3.3 X W Kainbow trout (largest) 7,(1 >' I0-" 7.0 X 10< Largenioulh bass (highest 2.3 X 10" 2.3 X 10= residues) Bay and Lakt Erie ZDUT Water 6.9 X 10-12 Sediment 1.7 X 10-8 2.5 X 10^' Bluegill (lowest 3.1 X 10-'- 4.5 X 10» residues) Coho salnmn ( largest ) 2.3 X 10-» 3.3 X lO"' Dieldrin Waler 1.5 X 10-1' Sediment 3.0 X 10-in 2.0 X W White bass (large) 1 .7 X 10-» 1.1 X 10= Creek and Lake Fish Among the fish caught in both the creeks and the bay, the plankton-feeders, rock bass (Amhloplites rupestris) and pumpkinseed (Lepomis gihbosm), had residues 10 to 1 5 times higher in those members caught in the creek than in those caught in the bay or lake (Fig. 4.5). The bottom-feeders, brown bullhead (Ictalurus nehiilosns) and carp {Cypriniis carpio), contained resi- dues of similar differences in magnitude. The marsh- feeding bowfin (Ainia calya) had low residues of both DDT and dieldrin. No true piscivore was caught in both the creek and lake waters for comparison. Discussion From the data collected it is possible to prepare an inventory of -DDT present in the four watersheds and the amount being slowly removed in agricultural pro- duce and by natural processes. The soil that acts as a reservoir was estiinated to contain 324,840 kg -DDT and 14,515 kg dieldrin in 1971. Over the past 11 years (1961-71) it was estimated that 882,730 kg DDT was added to the soil. From these data it was proposed that the half-life was between 3 and 4 years. Data compiled by Edwards showed that at B. Crappie C. Salmon A. Smelt S.M. Bass N. Redhorse Y. Perch W. Bass F. Drum G. Sunfish Alewtfe r Bluegill -t Pumpkinseed- -i Carp 1 W. Sucker S, Shiner C. Chub B. Trout C. Mudmin R. Trout B. Dace L.M. Bass BAY AND LAKE FISH a CREEK FISH 0.05 010 DIELDRIN^Ippml 0,15 0 20 FIGURE 5. Dieldrin concentrations in tissues of 23 fish species caught in creek and lake environments, southern Ontario — 1971 Vol. 8, No. 3, December 1974 199 dosages between I. II and 2.80 kg/ha., disappearance of 95 percent occurred in 4 to 30 years depending on soil, weather, and location (13). In the present study 95 percent disappearance was predicted to occur in 15 to 18 years, well within the span reported by Edwards. The average percent of parent DDT reported by Ed- wards that remained after 3 years was about 50 per- cent; however, his range was from 26 to 78 percent. In this study the figure was close to 80 percent. Mean residiies of -DDT found in soil agreed closely with the findings of Harris and Sans, in which the averages of four farms taken in 1964, 1966, and 1969 amounted to 3.06, 4.56. and 3.38 ppm i:DDT, respec- tively (5). In the present survey, soils recently cropped with tobacco contained mean residues between 3.88 and 4.36 ppm for each of the four watersheds. At the end of a 2- to 3-year rotation the same soils had mean residues ranging from 2.33 to 3.44 ppm. Composition of DDT at the beginning and end of the tobacco/ grain rotation showed little change, making it difficult to attribute disappearance to degradation. In woodlots, considerable degradation of DDT to DDE was evident (Table 3) compared to rye and tobacco fields. Air did not appear to account for volatile losses from the soil. Instead, some samples indicated that most of the DDT was derived from spray drift and others indicated that particulate matter was transported by air. In the annual marketing of tobacco, milk, and meat from the four watersheds, a total of 53.5 kg i;DDT and 2.13 kg dieldrin were removed. This represented 0.18 percent of the pesticide applied, assuming it was all from the 1971 application of DDT, which is doubt- ful. It is significant, however, that both the -DDT and the dieldrin removed by tobacco represented 0.035 per- cent of that resident in the soil used to produce the 1971 crop. Residues in hay and silage appeared to come from either spray drift or the soil and amounted to 38.3 kg :DDT and 0.76 kg dieldrin. Amounts found in meat and milk were calculated to be only 0.95 kg i^DDT and 0.16 kg dieldrin, a fraction of that found in the silage and hay used as animal feed. The average residue level in milk fat from the four watersheds. 0.188 ppm, was higher than the Provincial average of 0.134 ppm reported by Frank et al. from surveys conducted between 1967 and 1969 (10). The same authors have recently determined residue levels in milk from counties in the Lake Erie watershed (1). In surveys which they conducted in 1968-69 and 1970-71, Frank et al. discovered mean residues of 0.186 and 0.122 ppm i:DDT and 0.041 and 0.035 ppm dieldrin. respectively (14). Because tobacco production was the only major area of agriculture still using DDT in 1970-71, milk from counties in the tobacco belt, unlike milk from other counties in the Lake Erie watershed, did not show a decline in -DDT residues. Total annual removal of -DDT and dieldrin dissolved in water was estimated at 5.297 g I'DDT and 354.8 g dieldrin. Data collected by the 1971 Water Survey of Canada on Big Otter and Big Creeks determined that the amount of suspended sediment carried into Lake Erie by these two creeks was 54.4 and 13.4 x 10" kg/ year (//). Bed sediments from Big Otter Creek aver- aged 41.3 ppb i:DDT and 2.1 ppb dieldrin; those from the lower reaches of Big Creek averaged 38.7 ppb -DDT and 1.0 ppb dieldrin. If it can be assumed that suspended material contained residues similar to those of bed sediments, then it can be estimated that 1,600 g -DDT and 1 14 g dieldrin could have been discharged from Big Otter Creek and 393 g i;DDT and 13 g dieldrin could have been discharged by Big Creek into Lake Erie on suspended material. In other studies finer sediments tended to contain residues of -DDT and dieldrin higher than those of coarser sediments (7); therefore, it might be reasonable to predict that sus- pended material may carry higher and not lower resi- dues than bed sediments. Because no data were available on the movement of suspended solids from Nanticoke and Dedrich Creeks, no estimate of sediment residues can be made. Figures from the present study were only slightly higher than those of Miles and Harris (6). showing close agree- ment. In the case of Nanticoke Creek, sediment at the town of Nanticoke was only 10 ppb; hence the load carried by this creek is probably small. On Dedrich Creek residues in sediment were high. Because the creek is sluggish and the discharge volume is small, the amount of -DDT and dieldrin carried on suspended material is probably small also. Residues leaving the four watersheds in water or on suspended material can be estimated to be 7.3 kg/year -DDT and 0.5 kg/year dieldrin. This represents a loss of 38 and 2.6 mg/ha./year for DDT and dieldrin on a total watershed basis. If these losses are confined to the 40,455 ha. tobacco soil in the watersheds, then the amounts lost are 180 and 12 mg/ha./year for DDT and dieldrin, respectively, and represent a loss of 0.002 percent DDT and 0.003 percent dieldrin from the soil reservoir. In 1971 Miles and Harris reported that for the pre- ceding year the average weekly delivery of organo- chlorine insecticide from Big Creek into Lake Erie had been 50 g (6). This represents 2,600 g DDT/ year. This discharge included suspended matter as well as dissolved pesticide. In the present study conducted in 1971, it was calculated that 2.872 g iDDT and 130 g dieldrin were discharged into Lake Erie each year from Big Creek. Riverbed shift and suspended sediments could have car- ried an additional estimated 393 g -DDT and 13 g dieldrin into Lake Erie (Table 14). These figures from the present study are only slightly higher than those from Miles and Harris (6), showing close agreement. 200 Pesticides Monitoring Jouiwal The mean residues in water from Big and Dedrich Creeks were 20.0 and 14.9 ppt i;DDT and 0.9 and 1.1 ppt dieldrin. In Long Point Bay close to the outlets of these two creeks, residues in water were 12.5 ppt -DDT and 0.3 ppt dieldrin. Three miles off the mouth in Long Point Bay i;DDT residues were 1.3 ppt and dieldrin could not be detected. At the edge of Inner Long Point and Outer Long Point Bays, residues of -DDT were down to a trace. A marked dilution of residues occurred in water passing from creek to bay to lake. Mean residues in sediment were 38.7 and 141 ppb -DDT and 1.0 and 0.7 ppb dieldrin from the lower reaches of Big Creek and from Dedrich Creek. Close to shore in Long Point Bay the mean residLies were 28.7 ppt -DDT and 0.6 ppt dieldrin; in the middle of Long Point Bay sediments contained only 4.3 ppb 2DDT and no dieldrin. These data also illustrate dilution. Fish species caught in the creeks had markedly higher residues than those caught in the lake. Tissue levels in 10 species caught in the lake ranged from 0.043 to 2.38 ppm -DDT compared to tissue levels of 0.01 to 3.86 ppm -DDT in eight species caught in the creeks. Tissue levels of dieldrin ranged from nondetectable to 0.024 ppm in 10 species from the lake and 0.004 to 0,190 ppm for eight species from the creeks. These data emphasized differential accumulation of -DDT and dieldrin according to contamination of the location and the species involved. It is significant that of the six species caught in both systems, those members caught in the creeks had much higher residues than those caught in the lake. In both creek and lake, magni- fications of -DDT and dieldrin from water to the highest concentrations in fish tissue were of the order of lO-"'. The -DDT and dieldrin carried to Lake Eric in the four watersheds represented only 0.003 percent and 0.004 percent of those resident in the soil in 1971. The presence of this amoLmt in the aquatic environment was reflected by the higher residues in fish caught in the creek than in the lake. However, the level of -DDT in fish tissue remained below the 5 ppm tolerance per- mitted in commercial fish. Acknowlcilgiuenls Authors wish to thank H. Hoggarth and B. McAllister, Ontario Ministry of the Environment, for collecting air. water, soil, and sediment samples; E. H. Smith, Ontario Ministry of Agriculture and Food, for collecting milk; and members of the Ontario Ministry of Natural Re- sources for collecting fish. Thanks to G. Kurys. Ontario Ministry of the Environment, for his involvement in air monitoring. We gratefully acknowledge the assistance of Mrs. G. E. Bradshaw and J. W. McWade, Ontario Ministry of Agriculture and Food, for the extraction and cleanup of samples before determination, and that of R. Boelens. Ontario Ministry of the Environment, for sample collections from Long Point Bay. LITERATURE CITED ( 1 ) Agriciilliiral Statistics for Ontciiio. 1971 . Publication 20, prepared by the Statistics Section, Economics Branch, Ontario Ministry of Agriculture and Food. (2) Harris. C. R.. G. F. Manson. and J. H. Mazurek. 1962. Development of insecticidal resistance by soil insects in Canada. J. Fcon. Hntomol. 55:777-780. (3) Harris, C. R.. and H. J. Svec. 1968. Toxicological studies on cutworms. 1. Laboratory studies on the toxicity of insecticides to the dark-sided cutworm. J. Econ. Entomol. 61:788-793. (4) Harris. C. R.. H. J. Svec. and W. W . Sans. 1968. Toxicological studies on cutworms. II. Field studies on the control of the dark-sided cutworm with soil insecticides. J. Econ. Entomol. 61 :961-965. (5) Harris, C. R., and W. W. Sans. 1971. Insecticide resi- dues in soils on 16 farms in southwestern Ontario — 1964. 1966, and 1969. Pestic. Monit. J. 5(3 ) :259-267. (6) Miles. J. R. W.. and C. R. Harris. 1971. Insecticide residues in a stream and a controlled drainage system in agricultural areas of southwestern Ontario, 1970. Pestic. Monit. J. 5( 3 ) :289-294. (7) Frank, R., A. E. Armstrong, R. G. Boelens, H. E. Braiin, and C. W. Douglas. 1974. Organochlorine in- secticide residues in sediment and fish tissue, Ontario, Canada. Pestic. Monit. J. 7(3/4) : 165-180. (8) Langlois, B. E., A, R. Stemp, and B. J. Liska. 1964. Analysis of animal food products for chlorinated in- secticides. J. Milk Food Tech. 27:202-204. (V) Hamenee, H. H., P. S. Hall, and D. J. Caverley. 1965. The identification and determination of chlorinated pesticide residues. Analyst 90:649-656. UO) Frank. R., H. E. Braun. and J. W . McWade. I97U. Chlorinated hydrocarbon residues in the milk supply of Ontario, Canada. Pestic. Monit. J. 4(1):3I-4I. ( / / ) Water Survey of Canada, Inland Waters Directorate, Environment Canada. Dec. 31, 1973. Surface water and sediment data for Canadian rivers, 1965-71. (/2| Berst, A. H., and H. R. McCrimmon. 1966. Compara- tive summer limnology of Inner Long Point Bay, Lake Erie, and its major tributary. J. Fish Res. Board Canada 23:275-291. (/.') Edwards, C. A. 1966. Insecticide residues in soils. Residue Reviews 13:83-132. (14) Frank, R., E. H. Smitli, H. E. Biaun, M. Holdrinet, and J. W. McWade. 1975. Organochlorine insecticides and industrial pollutants in the milk supply of the Southern Region of Ontario, Canada. J. Milk Food Technol. (in press). (/5) .Annual Report. July 19, 1972. Ontario Flue-Cured Tobacco Growers Marketing Board, Tillsonburg, Ontario. il6) Unpublished Records. 1971. Ontario Milk Marketing Board, 50 Maitland Street, Toronto. (/7) Annual Livestock .Market Review. 1971. Ontario cattle marketings by counties, 72:43-44. Markets Infor- mation Section, Production and Marketing Branch, Agriculture Canada, Ottawa. (/iS) Clinuitological Station Report. 1971. Research Station, Agriculture Canada, Delhi, Ontario. Vol. 8, No. 3, December 1974 201 Persistence and Movement of BHC in a Watershed, Mount Mitchell State Park, North Carolina— 1967-72 ' M. D. Jackson,2 T. J. Sheets,3 and C. L. Moffett * ABSTRACT An experimental area in Mount Mitchell State Park, North Carolina, was sprayed with BHC, benzene bexaehloride, at an average rate of 1 1.2 kg/ha. to control the balsam woolly aphid (Adelges piceae). Residues were 31 and 585 ppm in soil and litter, respectively, I month after spraving. A high percentage of the residue appeared to remain in the treated area; contamination of streams draining the area was minimal. BHC present in surface litter after applica- tion slowly moved into surface soil. The residue level in surface soil reached a high of 58 ppm 1.5 years after applica- tion. At this time the concentration in litter averaged 134 ppm. Residues in soil and litter were 32 and 27 ppm, re- spectively, 5 years after application. Concentrations of BHC in animals were unrelated to trapping locations. Introdiiclinn In North Carolina infestations of balsam woolly aphid {Adelges piceae) were first observed in 1957 among stands of Fraser fir (Ahies frascri) in the western North Carolina mountains (/). Since that time stands have been severely reduced in many areas; in some areas all mature trees have been killed. Seedlings devel- oping in such areas are also attacked and succumb within 1 to 7 years (2). Except for commercial plantings for Christmas tree production, the value of Fraser fir is primarily, if not ' Journ.il Series No. 414S, North Carolina .State tiniver-iity Agriciil- liiral Experiment Station Supported in part by Division of Slate Parks. North Carohna Department of Conservation and Develop- ment (now Nt>rth Carolina Department of Natural and Economic Resources) . -Former employee of Pesticide Research Laboratory. North Carolina State University. Raleigh. N C. Present address: Pesticides and Toxic Substances Effects Laboratory (NERO, U.S. Environmental Protec- tion Agency. Research Triangle Park, N.C. ^ Professor. Pesticide Research Laboratory, North Carolina State L'ni- versity. Raleigh. N.C. 27607. Reprints available from this address. ^ Former employee of North Carolina Department of Conservation and Development. Division of State Parks, Raleigh. N.C. Present address: Division of Refuges, Bureau of Snort Fisheries and Wildlife, U.S. Fish and Wildlife Service. Washington, D.C. entirely, aesthetic. The tree occurs naturally in North Carolina only at altitudes above 1200 m. Both lime sulfur (calcium polysulfides) and BHC (1 .2.3,4,5.6-Hexachlorocyclohexane) have been used to control the balsam woolly aphid. Because lime sulfur has no residual activity, retreatment for several years is required for effective control. Therefore, control with lime sulfur is more expensive than it is with BHC. However, the potential for accumulation in nontarget biota is greater with BHC. Several investigators have studied the depletion of BHC and lindane, the gamma isomer of BHC. from soil. Using electron-capture/gas-liquid chromatography (EC/GLC), Nash and Woolson (.?) found an average residue of 10 percent from two BHC applications of 56 and 224 kg/ha. 14 years later. Lichtenstein and Polivka (4) recovered an average of 41 percent by a colorimetric method 1 1 years after application of 0.3, 2.S, 5.6, and 1 1 kg/ha. BHC. In another study Lich- tenstein et al. (5) detected 0.2 percent lindane, origi- nally applied at rates of II and 112 kg/ha.. 15 years later. Although BHC and lindane are less persistent than most other chlorinated hydrocarbon insecticides (3,5). residues remained for several years in soils. Longevity was influenced by soil type, moisture level in soil, evap- oration of water from soil, temperature, and other fac- tors (6.7.8). Soil microorganisms appear to be involved in decomposition of lindane (9) and other isomers of BHC (10) in submerged soils. The residual properties of BHC which contribute to its long-term control of the balsam woolly aphid may be undesirable from the standpoint of nontarget biota. The investigation reported here was undertaken to study the persistence of BHC in a typical Fraser fir stand. In 202 Pesticides Monitoring Journal conjunction with a spray operation on the north side of the Mount Mitchell peak in 1967, experimental plots and sampling schemes were established for sev- eral reasons: to study the deposition and persistence of BHC in surface litter and soil within the sprayed area; to determine residues in water and sediment of small streams within and below the treated area; and to measure residues in small mammals in sprayed and unsprayed areas. Experimental Procedures Three 0.04-ha. plots were marked in Mount Mitchell State Park, North Carolina, on the north side of the peak at an elevation of about 1900 m. Plot I contained an intermediate stand of Fraser fir; plot II, a relatively young stand of Fraser fir; and plot III, an old stand of Fraser fir. BHC was applied to a 13-ha. area on the north side of the mountain June 28 — August 30. 1967. Each tree was sprayed individually with a 1:100 dilution of BHC con- centrate containing 11.1 percent of the gamma isomer and 16.7- percent of other isomers. The 13-ha. area was sprayed at an average rate of approximately 1 1.2 kg/ha. BHC. The three experimental plots lay within the sprayed area. Plot III was sprayed June 28, 1967; plots I and II were sprayed July 5 the same year. Surface litter was defined as the organic material above, but not including, the 02 horizon (//). A sample con- sisted of litter from 20 random sites (grab subsamples) within each experimental plot. Each plot was sampled twice on each sampling date. Researchers randomly took two 20-core samples of soil, each core 2.5 cm in diameter and 15 cm deep, from all three plots several times during a 5-year period after application. Because the area treated in 1967 had been sprayed with BHC, control samples of soil and litter were taken from an unsprayed area at Indian Gap in the Great Smoky Mountains. Water samples were collected several times during a 2-year period from site 1, a small stream immediately below plot II and within the sprayed area. Site 2 was established below the treated area in a small stream which had been formed by drainage from the treated area. Because the streams were very small, the entire cross section of the stream was sampled. On each sam- pling date, three 3.8-liter samples were taken from each site. The water was stored at 3°-5°C in brown glass jugs. Samples were usually transported to the laboratory on the same day and extracted on the fol- lowing day. Water was not filtered before extraction. Grab samples of sediment were taken from the same sites. Each sample was a composite of sediment from five locations 2 to 5 cm deep along an 8-m length of the stream. Field personnel took three samples at each sampling site. Small mammals were caught periodically in snap-traps within each plot and outside the treated area during a 3-year period after spraying. During trapping periods, trap lines were run at 12-hour intervals. The type and number of animals were limited; therefore, all trapped animals were sacrificed and analyzed for BHC residues. All samples except water were stored at — 18°C until analysis. The extraction method for water was identical to that used by Bradley et al. {12). Soil and sediment were air-dried and passed through a No. 18 sieve. Sub- samples of 100 g each were extracted by a procedure adapted from one used in the National Soils Monitor- ing Program (13,14). Forest litter samples were extracted by the aqueous- acetonitrile method for low-moisture and low-fat sam- ples (15). Animals of the same species which weighed less than 40 g were grouped by areas. Animals weighing more than 40 g were analyzed separately. The entire animal, including skin and digestive tract, was analyzed. A frozen animal sample was chopped in a Hobart food chopper with sufficient anhydrous sodium sulfate to absorb the moisture. The entire sample was transferred to 500-ml centrifuge bottles. Each bottle was extracted three times with petroleum ether (200, 100, and 100 ml, respectively). The bottles were centrifuged at 1500 rpm for 5 minutes and the solvent layer was decanted through anhydrous sodium sulfate. Extracts were com- bined, and the solvent was evaporated with a stream of dry air. Fatty materials remained. A 3-g aliquot of animal fat was subjected to the stan- dard acetonitrile partitioning procedure (15). Extracts from the acetonitrile partitioning of the fat and the litter extract were chromatographed on an activated florisil column; BHC was eluted with 6 percent diethyl ether in petroleum ether (15). Water, soil, and sediment extracts did not require cleanup before gas chroma- tography. The alpha, beta, and gamma isomers of BHC were determined with a model MT-220 gas chromatograph equipped with a ^'Ni electron-capture detector. A 183- cm-by-0.3-cm, U-shaped, glass column packed with 6 percent QF-1 and 4 percent SE-30 on Gas Chrom Q (60/80 mesh) was used with a nitrogen flow rate of 100 cc/min. Column, injection port, and detector tem- peratures were 175°, 220°, and 250°C, respectively. All gas chromatographic measurements were made by the peak height method; amounts present were calcu- lated against standards run daily. Recoveries of added Vol. 8, No. 3, December 1974 203 amounts of BHC fror.\ soil, fat, litter, and water are in Table 1. Results ami Discussion Rainfall and temperature data were taken near the experimental area at the same elevation (Table 2). These data show that the weather was typical for the high mountainous area of the Southeastern United States. Sampling sites received little or no direct sun and the area was cool and damp, even in summer months. The experimental area had been sprayed with BHC at approximately 11 kg/ha. in 1963. Litter and soil sam- ples collected before the 1 967 application contained BHC residues of 67 and 17 ppm, respectively (Tables 3.4). Initially, the forest litter contained the highest level of BHC. The concentration in the litter averaged 585 ppm about 1 month after application; residues decreased gradually over the next 5 years to 27 ppm. Because pretreatment BHC levels in soil averaged 1 7 ppm, the initial deposit in the surface soil from the 1967 spraying was 31 minus 17 ppm, or about 14 ppm: surface soil was usually but not always beneath a layer of litter. BHC residues in soil increased to a peak of 58 ppm 1.5 years after application. Thereafter, the concentration slowly declined to 32 ppm 3.5 years later. Although no specific weights of soil and litter layers were recorded, authors do know that the 1 5-cm soil layer weighs several times as much as the overlying litter. Therefore, the increase of BHC concentration in the soil from about 31 ppm at the first sampling date after application to the peak concentration of 58 ppm 1.5 years later may account for 50 percent or more of the BHC lost from the litter (585 ppm to 134 ppm) during this same period. Extended persistence of the insecticide in this soil probably can be attributed to conditions unfavorable to microbial activity: highly acid soils, cool soil temperatures most of the time, and low intensity of solar radiation within the forest area (3.10). The present data on persistence of BHC in soil are in general agreement with those of other authors who showed that BHC persisted at least 14 years in soils of a climate more moderate than the mountains of North Carolina (3.5). Residues in the water from site 2, which drains the entire watershed, were usually below the limit of de- tection (0.06 ppb:Table 5). On August 30, 1967, after several heavy rains between August 19 and August 27 totaling 18 cm, the stream flow was above normal. Insecticide spraying had been completed by August 30, and the BHC level in samples from the stream aver- aged 6.3 ppb. April 16, 1969, was the only other sam- pling date on which there was an appreciable amount of BHC in the stream below the sprayed area. TTiis sampling was made during runoff of melting snow. Water collected from the small stream within the treated area usually contained low concentrations of BHC (Table 5). Sediment from site 1 , the stream within the treated area, contained residues from 0.49 to 3.17 ppm (Table 6). Residues in sediment from site 2, the stream below the treated area, ranged from <0.03 to 0.21 ppm. Total residues in the fat of animals taken from treated areas ranged from 0.6 to 106 ppm, and those in fat of animals from outside the treated area ranged from 0.1 to 176 ppm (Table 7). There was no relation between BHC residues in animals and the trapping area (treated versus untreated): therefore, no animals were sampled after 1969. Most animals sampled in 1967 and 1968 TABLE 1. Recoveries of BHC isomers from animal fat, forest litter, soil or sediment, and water Average Recovery Type BHC No. AMOUNT RECOVERY, RANGE, Sensitivity SAMPLE ISOMER SAMPLES ADDED ^ % % LEVEL 1 Animal Alpha 8 0.1-100 92 87- 95 0.02 fat Beta g 0.3-300 92 82- 97 0.06 Gamma 8 0.1-100 91 84- 95 0.02 Forest Alpha 15 0.2-40.0 99 80-112 0.01 litter Beta 15 0.2-200 98 81-110 0.01 Gamma 15 0.2-200 103 82-142 0.01 Soil or Alpha 18 0.05-10.0 98 82-127 0.01 sediment Beta 18 0.05-20.0 94 54-136 0.01 Gamma 18 0.05-20.0 97 78-126 0.01 Water Alpha 8 0.02-1.0 80 65- 91 0.02 Beta 8 0.02-5.6 93 73-148 0.02 Gamma 14 0.02-2.0 82 70- 90 0.02 NOTE: BHC was added immediately before extraction. ^ Units are ppm for animal fat, forest litter, soil, and sediment, and ppb for water. 204 Pesticides Monitoring Journal and all those sampled in 1969 were from outside the treated area, although they normally travel across treated and some untreated portions of the area. Attempts to trap animals in the treated area wore unsuccessful, and no dead animals were found during routine searches. AckiiowleclgmeiUs Authors thank K. W. Greenlee, J. R. Wilson, and B. J. Taylor, Division of State Parks, North Carolina Depart- ment of Natural and Economic Resources, for their assistance in conducting this study. LITERATURE CITED (/) Speers, C. F. 1958. The balsam woolly aphid in the southeust. J. Forest 56(7) :515-5 16. (2) Cieshi, W. M., H. L. Lambert, and R. T. Franklin. 1965. Status of the balsam woolly aphid in North Carolina and Tennessee. 1964. USDA Forest Service, Report 65-1-1. 10 pp. (3) Nash, R. G., and E. A. WooLson. 1967. Persistence of chlorinated hydrocarbon insecticides in soils. Science 157(3791 );924-927. (4) Lichtenslein, E. P., and J. B. Polivka. 1959. Persis- tence of some chlorinated hydrocarbon insecticides in turf soils. J. Econ. Entomol. 52(2)289-293. (5) Lichtenstein, E. P., T. W . Fuhremann, and Kenneth R. Schidz. 1971. Persistence and vertical distribution of DDT, lindane, and aldrin residues, 10 and 15 years after a single soil application. J. Agr. Food Chem. 19(2):718-721. (6) Bowman, M. C, M. S. Schechler, and R. L. Carter. 1965. Behavior of chlorinated insecticides in a broad spectrum of soil types. J. Agr. Food Cheni. 13(4): 360-365. (7) Lichtenstein, E. P., and K. R. Schidz. 1959. Persistence of some chlorinated hydrocarbon insecticides as influ- enced by soil types, rate of application and tempera- lure. J. Econ. Entomol. 52( 1 ): 124-13 1. (8) Yide, W. N.. M. Chiba, and H. V. Morley. 1967. Fate of insecticide residues. Decomposition of lindane in soil. J. Agr. Food Chem. 15(6) : 10(10-10(14. (9) Raghii, K., and L C. MacRae. 1966. Biodegradation of the gamma isomer of benzene hexachloride in sub- merged soils. Science 154(3746 ) :263-264. (10) MacRae. I. C, K. Raghii, and T. F. Castro. 1967. Persistence and biodegradation of four common isomers of benzene hexachloride in submerged soils. J. Agr. Food Chem. 15(5 ) :yi 1-914. (//) Bnol. S. W., F. D. Hole, and R. J. McCracken. 1973. Soil genesis and classification. The Iowa State Univer- sity Press. Ames, Iowa. 360 pp. {12} Bradley, J. R., Jr., T. J. Sheets, and M. D. Jackson. 1972. DDT and toxaphene movement in surface water from cotton plots. J. Environ. Quality 1 ( 1 ): 102-105. (13) Sheets, T. J., M. D. Jackson, W. J. Mislric, and W. V. Campbell. 1969. Residues of DDT and dieldrin in pea- nuts and tobacco grown on contaminated soil. Pestic. Monit. J. 3(2):80-86. (14) U.S. Department of Agriculture. 1966. Monitoring agricultural pesticide residues. ARS 81-13. U.S. Gov- ernment Printing Office, Washington, D.C. (15) Burke, J. A., J. A. Gaul, and P. E. Corneliussen. 1971. Pesticide Analytical Manual Vol. I, U.S. D HEW, Wash- ington, D.C. Vol. 8, No. 3, December 1974 205 TABLE 2. Cliniatologkal Jala near BHC experimental area. North Carolina— 1967-72 TABLE 3. BHC residues in forest litter, North Carolina — 1967-72 Temperature High Low Avo Rainfall, Year Month (°C) (°C) (°C) CM 1967 June 21.1 2.8 11.7 28.3 July 20,0 3.3 11.7 22.7 Aug. 20.0 2.2 11.1 19.9 Sept. 18.9 —6.1 6.1 11.2 Oct. 18.9 —5.6 6.7 12.1 Nov. l.V.l — 17.2 —2.2 10.5 Dec. 14.4 —16.7 —1.1 19.3 1968 Jan. 12.8 —18.9 —3.3 19.4 Feb. 10.0 —21.1 —5.6 1.8 Mar. 17.2 —17.8 —0.6 21.8 Apr. 20.0 —3.3 8.3 17.0 May 20.0 —3.3 8.3 14.1 June 24.4 3.3 13.9 31.7 July 21.1 2.2 11.7 10.3 Aug. 22.8 5.6 13.9 12.6 Sept. 20.0 3.3 11.7 9.4 Oct. 17.8 —8.9 4.4 32.6 Nov. 18..1 —14.4 1.7 12.0 Dec. 18.3 —14.4 1.7 12.7 1969 Jan. 8.9 —21.7 —6.7 13.1 Feb. 7.8 —17.8 —5.0 22.3 Mar. 7.8 —17.8 —5.0 19.4 Apr. 20.0 —7.8 6.1 20.3 May 21.1 —5.6 7.8 15.3 June 23.3 0 11.7 19.9 July 22.8 10.0 16.1 18.0 Aug. 20.0 6.1 12.8 18.2 Sept. 17.8 3.3 10.6 12.8 Oct. 21.1 —7.8 6.7 15.1 Nov. 13.3 —20.0 —3.3 24.3 Dec. 8.9 —14.4 —2.8 23.1 1970 Jan. 8.9 —31.1 —11.1 9.6 Feb. 14.4 —24.4 —5.0 12.0 Mar. 12.2 —17.8 —2.8 13.0 Apr. 19.4 —11.1 3.9 15.7 May 21.1 —1.1 10.0 8.7 June 21.1 3.3 12.2 12.3 July 21.1 5.6 13.3 20.9 Aug. 21.1 7.8 14.4 21.8 Sept. 22.8 —2.2 10.0 6.3 Oct. 16.1 —4.4 5.6 45.6 Nov. 12.2 —22.8 —5.6 14.8 Dec. 10.0 —20.0 —5.0 10.3 1971 Jan. 10.0 —18.9 — 4.4 16.8 Feb. 11.7 —18.9 —3.9 19.8 Mar. 13.3 —18.9 —2.8 16.3 Apr. 17.8 —7.8 5.0 11.5 May 20.0 —8.9 5.6 14.1 June 21.1 7.8 14.4 10.9 July 21.1 4.4 12.8 22.2 Aug. 20.0 7.2 13.3 24.0 Sept. 20.0 5.6 12.8 14.2 Oct. 18.9 —3.3 7.8 34.9 Nov. 15.6 —14.4 0.6 19.7 Dec. 15.6 —14.4 0.6 19.6 1972 Jan. 12.2 —12.2 0 21.7 Feb. 15.0 —17.8 —1.7 12.9 Mar. 14.4 —14.4 0 2C.5 Apr. 20.0 —12.2 3.9 11.3 May 15.6 —2.2 6.7 24.9 June 18.9 —2.2 8.3 33.4 July 21.1 3.3 12.2 20.2 Aug. 20.0 8.9 14.4 4.9 Plot BHC ISOMER Sampling DATE Alpha (PPM) Beta (PPM) Gamma (PPM) Total (PPM) 5/18/67 Check 1 0.04 0.01 0.01 0.06 5/19/67 4.68 11.9 4.78 19.1 75.9 40.4 9.00 26.4 10.1 32.78 114.2 55.37 7/7/67 6/30/67 145 88.6 119 298 188 302 252 158 205 696.0 434.6 626.0 8/2/67 98.2 71.2 46.0 254 226 152 188 142 91.4 540.2 439.2 289.4 9/1/67 66.5 44.4 60.0 256 148 210 142 95.4 126 464.5 287.8 396.0 10/31/67 60.3 35.8 47.4 262 150 238 130 80.8 106 452.3 266.6 391.4 5/23/68 74.9 44.5 73.5 236 170 286 188 101 178 498.9 315.5 537.5 6/25/68 42.6 33.8 22.8 182 134 98.7 102 79.4 48 326.6 247.2 169.5 11/6/68 17.2 13.5 19.2 91.5 56.2 83.4 41 32.1 47.9 149.7 101.8 150.5 8/20/693 19.6 17.8 18.7 73.0 58.2 71.0 41.1 33 40.6 133.6 109.0 130.3 10/29/70 6.46 4.86 8.33 11.2 9.22 19.2 39.4 30.4 58 57.06 44.48 85.53 7/19/72 4.07 2.14 3.62 20.3 10 22.0 7.38 3.88 7.20 31.75 16.02 32.82 NOTE: Residues expressed are as received. All values are averages of two samples except for those from check plot. 1 Average of three samples from Indian Gap, Great Smoky Moun- ■ tains. - Samples collected May 19, 1967, are pretreatment samples from experimental area. ^ Average of three samples. 206 Pesticides Monitoring Journal TABLE 4. BHC residues in soil, North Carolina— 1967-72 TABLE 5, Plot BHC ISOMER 1 Sampling DATE ALPHA (PPM) Beta (PPM) Gamma (PPM) Total (PPM) 5/18/67 Check ■ 0.01 0.01 0.08 0.10 5/19/67 1 = 2 3 0.81 1.50 4.72 3.36 6.36 10.6 1.82 3.46 17.2 5.99 11.32 32.52 7/6/67 6/30/67 1 2 3 4.15 3.48 4.69 14.2 13.4 27.6 8.34 6.72 9.79 26.69 23.60 42.08 8/2/67 1 2 3 1.44 4.44 2.89 4.56 12.9 12.1 2.80 7.59 6.14 8.80 24.93 21.12 9/1/67 2 3 4.82 5.54 5.26 13.3 16.0 18.2 8.44 10.8 10.7 26.56 32.34 34.16 10/31/67 1 2 3 4.03 4.18 4.72 16.4 14.8 17.6 9.33 7.77 9.40 29.76 26.75 31.72 5/23/68 2 3 5.21 3.77 7.34 15.7 14.6 30.6 10.6 8.68 16.0 31.51 27.05 53.94 6/25/68 1 2 3 4.50 2.53 6.66 14.7 10.9 25.4 8.50 5.92 15.2 27.70 19.35 19.35 11/6/68 1 2 3 10.3 7.58 7.42 34.6 26.6 25.8 21.9 21.8 17.6 66.80 55.98 50.82 8/20/69 ■' 1 2 3 8.68 3.84 6.09 17.3 9.43 12.4 17.7 8.03 13. T 43.68 21.30 32.19 10/29/70 1 2 3 3.72 3.22 8.16 13.0 12.0 22.6 11.7 11.7 24.1 28.42 26.92 54.86 7/19/72 1 2 3 2.74 3.42 5.30 11.1 20.2 28.6 6.16 7.27 11.7 20.00 30.89 45.60 BHC residues in water within and below treated area, North Carolina — 1967-69 NOTE: Residues expressed are oven-dry weight. Values are the average of two samples except as indicated. J- Average of three samples from Indian Gap, Great Smoky Moun- tains. 2 Samples collected May 19, 1967, are pretreatment samples from experimental area. ^ Average of three samples. BHC isomer Sampling Alpha Beta Gamma TOT/IL DATE Site " (PPB) (PPB) (PPB) (PPB) 5/18/67 Control <0.02 <0.02 <0.02 <0.06 5/19/67 1 = 0.04 <0.02 0.12 0.17 -) <0.02 <0.02 0.02 <0.06 7/ 6/67 I 1.14 1.13 2.63 4.90 2 <0.02 <0.02 0.03 <0.06 7/ 7/67 23 <0.02 <0.02 0.03 <0.06 7/29/67 2' <0.02 <0.02 <0.02 <0.06 8/ 1/67 23 <0,02 <0.02 <0.02 <0.06 8/ 2/67 1 0.86 1.82 2.11 4.79 2 <0.02 <0.02 <0.02 <0.06 8/22/67 1 3.57 4.18 8.67 16.42 2 <0.02 0.04 0.03 0.08 8/30/67 1 <0.02 <0.02 <0.02 <0.06 2 1.15 2.42 2.70 6.27 10/31/67 1 0.06 0.10 0.16 0.32 2 <0.02 <0.02 <0.02 <0.06 5/23/68 1 0.21 2.75 1.71 4.67 2 <0.02 <0.02 <0.02 <0.06 6/25/68 1 0.75 5.04 2.28 8.07 2 <0.02 0.03 <0.02 <0.06 11/ 6/68 2 <0.02 <0.02 <0.02 <0.06 4/16/69 1 0.92 2.43 2.18 5.52 2 0.19 0.28 0.27 0.74 NOTE: Values are averages of three samples analyzed separately ex- cept as indicated. 1 Site 1 is within treated area; site 2 is below. ' Only two samples analyzed. ^ Values are averages of three samples collected during a rainstorm on each date. TABLE 6. BHC residues in sediment within and below treated area. North Carolina — 1967-69 BHC isomer Sampling Alpha Beta Gamma Total DATE Site (PPM) (PPM) (PPM) (PPM) 7/ 6/67 1 2 0.34 0.01 2.00 0.02 0.83 0.10 3.17 0.13 8/ 2/67 9/ 1/67 2 1 2 0.07 <0.01 0.07 0.01 0.71 0.01 0.46 0.03 0.16 <0.01 0.15 0.01 0.94 <0.03 0.68 0.05 10/31/67 5/23/68 2 1 -> 0.01 0.06 0.01 0.06 0.42 0.18 0.01 0.16 0.02 0.08 0.64 0.21 6/25/68 1 2 0.04 0.02 0.34 0.05 0.11 0.02 0.49 0.09 11/ 6/68' 2 0.02 0.03 <0.01 0.06 NOTE: Residues expressed are oven-dry weight. Values are the aver- age of three samples except as indicated. 1 Average of two samples. Vol. 8, No. 3, December 1974 207 TABLE 7. BHC residues in animal fat, North Carolina— 1967-69 Number and Lipid, BHC ISOMERS Sampling Alpha Beta Gamma Total YEAR Location TYPE OF animal 1 % (PPM) (PPM) (PPM) (PPM) 1%7 Plot I 3 Deer mice 12.6 0.70 1.20 0.30 2.20 1 Little brown myotis bat 1.4 0.63 2.93 0.86 4.42 3 Deer mice 7.9 36.4 42.2 27.5 106.1 1 Boreal redb^ck vole 5.8 0.37 n.55 0.27 1.19 Plot II 1 Deer mouse 9.7 3.06 6.01 4.85 13.92 1 Boreal redback vole 7.6 0.15 0.19 0.25 0.59 Plot III 1 Deer mouse 7.4 0.32 0.60 0.57 1.49 1 Boreal redback vole 3.3 2.51 3.96 2.58 9.05 Outside 2 Deer mice 7.0 0.38 0.52 0.24 1.14 treated 1 Deer mouse 4.1 16.9 17.6 4.58 39.08 area 1 Dear mouse 5.9 0.20 0.78 0.31 1.29 3 Deer mice 8.2 0.51 0.67 0.43 1.61 I Boreal redback vole 3.3 0.11 0.45 0.20 0.76 1 Boreal redback vole 2.9 0.09 0.34 0.18 0.61 1 Boreal redback vole 5.7 0.07 0.27 0.12 0.46 1 Boreal redback vole 6.3 0.11 0.18 0.28 0.57 2 Boreal redback voles 6.3 61.4 29.6 56.8 147.8 1 Boreal redback vole 2.9 36.0 127. 12.5 175.5 1 Shorttail shrew 2.4 0.27 0.84 0.51 1.62 3 Shorttail shrews 8.2 11.9 33.6 13.3 58.8 1 Woodland jumping mouse 3.0 0.23 0.47 0.35 1.05 1 Woodland jumping mouse 1.4 5.23 24.2 7.10 36.53 1968 Plot I 3 Deer mice 13.8 1.55 2.98 0.97 5.50 3 Deer mice 16.2 0.98 1.38 0.71 3.07 3 Deer mice 19.8 0.69 0.83 0.38 1.90 1 Boreal redback vole 8.8 9.33 2.94 3.00 15.27 1 Boreal redback vole 33.4 1.25 1.54 0.69 3.48 Outside 3 Deer mice 10.2 1.24 1.67 0.90 3.81 treated 3 Deer mice 13.2 0.10 0.16 0.08 0.34 area 3 Deer mice 11.3 0.20 0.25 0.11 0.56 1 Boreal redback vole 8.1 0.03 0.11 0.03 0.17 1 Boreal redback vole 15.3 0.02 0.07 0.02 0.11 1 Boreal redback vole 3.5 3.46 3.91 1.29 8.66 1 Norway rat 6.5 0.45 0.14 0.20 0.79 1 Woodchuck 4.8 0.29 0.41 0.19 0.89 1969 Outside 2 Deer mice 11.7 0.09 0.34 0.21 0.64 treated 2 Deer mice 13.4 <0.02 <0.06 0.02 <0.10 area 1 Boreal redback vole 13.0 0.02 0.06 0.02 0.10 1 Boreal redback vole 19.2 0.05 0.24 0.02 0.31 NOTE: Residues expressed as extractable lipids. * Deer mouse, Peromyscus numiculatus; little brown bat, Myotis lucifugiis; Boreal redback vole, Clethrionomys gapperi; short-tailed shrew, BUirtna brevicauda; woodland jumping mouse, Napueozapus insignis; Norway rat, Rallus norvegicus; and woodchuck, Marmota monax. 208 Pesticides Monitoring Journal Contribution of Household Dust to the Human Exposure to Pesticides * Herbert G. Starr, Jr.,- Franklin D. Aldrich,' William D. McDougall III,' and Laurence M. Mounce ■ ABSTRACT Preliminary analysis of environmental contributions to pesti- cide body burden revealed household dust as a major reser- voir of pesticides in the environment. A year-long monthly study of the households of pesticide-exposed persons and control households in Weld County, Colo., in 1968 revealed appreciable levels of selected chlorinated pesticides in the exposed group. Exposed subjects varied from entire farm families with high agricultural use of pesticides to house- holds with at least one member who formulated pesticides, either as an employee of a local plant or as a professional applicator who mixed and loaded pesticides for commercial use. In the overall data no quantitative relationships were demonstrated between pesticide levels in household dust and pesticide levels in blood, although circumstantial data from individual households indicate that a certain connection does exist. No correlation could be shown between levels of p,p'-DDT and p.p' -DDE in household dust. Pesticide levels in the dust indicate a probable influence on body burden contributing to total environmental exposure of the individ- ual to pesticides. Introdiiclion Pesticide body burden in the general population can be augmented by dust, air (I), water, food, and the nature of the home and working environments. Differences in pesticide levels distinguishing the general population from a pesticide-exposed population may then be attrib- uted to occupational exposure and to pesticides carried into homes on clothing and other items. Surveys of ' Research supported by contract PH-86-65-62 witli ttie Division of Community .Studies, Atlanta, Ga. Formerly part of Food and Drug Administration. U.S. Department of Healtli. Education, and Welfare. Presently under Technical Services Division. Office of Pesticide Pro- grams. U.S. Environmental Protection Agency, Washington. D.C. 2 Former employee of Colorado Epidemiologic Pesticide Study Cen- ter, Colorado Slate University, Fort Collins, Colo. 8052.1. (Reprints available from this address.) * Environmental Medical Service, Massachusetts Institute of Tech- nology, Cambridge, Mass. * Denver Police Forensic Laboratory, Denver, Colo. pesticide content in food, soil, and water have been con- ducted for many years without attempting to relate levels found to pesticide body burden. The present study was initiated to determine the relationship between pes- ticide levels in household dust and pesticide levels in human blood sera in Weld County. Colo., which has a high annual usage of pesticide chemicals because it is primarily an agricultural area. Preliminary analyses of soil, water, and household dusts conducted in 1966 revealed pesticide levels to be higher in household dusts than in soil and water, and indicated some correlation between DDT levels in dust and DDT and DDE levels in sera of householders (2). In order to minimize the variables associated with different sam- pling times and seasonal peaks in pesticide use. the study was conducted for 1 year. From multiple regres- sion analyses and available data, the following hypoth- esis was formed: a definable relationship exists between blood serum chlorinated hydrocarbon pesticide levels and household dust levels. Reviews of published literature revealed no attempt to measure pesticide levels in household dusts. Modifica- tions of other soil assay methods were incorporated in the analytical method for this study to provide sensi- tivity and specificity needed for accurate analyses of pesticides in household dusts. Storm dust fall has been analyzed (3) for organochlo- rine pesticide levels using microcoulometric/gas-liquid chromatography (MC/GLC) in the quantitative step. Pesticide levels in soils have been studied by many re- searchers who have designed a variety of soil extraction and cleanup techniques for adaptation to current needs (4-6). This paper attempts to report results obtained from a year's study of pesticide levels in both house- hold dusts and blood sera, and, when possible, to dem- onstrate the relationship of pesticide levels between these two substrates. Vol. 8, No. 3, December 1974 209 Analytical Methods HOUSEHOLD DUST Household dust from a month's sweepings was collected in disposable vacuum cleaner bags and sifted. Particles passing through a Number 60 mesh screen were taken for analysis. One gram of air-dried dust was soxhlet- extracted with 1:1 hexane: acetone and cohimn-chro- matographed on deactivated aluminum oxide using hexane as the eluent to remove waxes and other ex- tractables. Final separation of chlorinated hydrocarbon pesticides was obtained by florisil column chromatog- raphy. Final concentrations of household dust extracts were 2.0 |^(g/fil for analysis by electron-capture/ gas- liquid chromatography (EC/GLC) and 1.0 mg/|.il for analysis by MC/GLC. An arbitrary sensitivity limit of 1.50 parts per million fppm) was established so that all residues could be qualitatively confirmed by MC/ GLC. Organochlorine insecticide recovery data for fortified household dust screenings are given in Table 1 . All recoveries were within acceptable analytical ranges. Relatively high standard deviations may have been caused by difficulty in obtaining uniform fortification rather than by the method. TABLE 1. Recovery of pesticides from fortified hoiisediist samples. Colorado — 1968 No. Mean Standard Compound Samples Recovery, % Deviation ■ Lindane 12 87.3 6.99 Heptachlor Epoxide 12 80.6 12.86 Dieldrin 11 86.. 1 12.89 fji'-DDT 9 S7.8 20.20 P.P'-DDE 12 82.9 6.35 P.P'-DDD 12 90.0 9.72 ' Relatively high standard deviations possibly caused by difficulty in obtaining uniform fortification rather than by method. BLOOD Blood samples were collected in 10-ml Vacutainer tubes containing no anticoagulant. After clotting, 2.0 ml serum was extracted with hexane using the method of Dale, Curley, and Cueto (7). Samples were analyzed by EC/GLC with an arbitrary lower sensitivity limit of 5 parts per billion ( ppb ) . GAS CHROMATOGRAPHY All analyses were performed on Micro-Tek 220 gas chromatographs equipped with H'' electron-capture de- tectors. Household dust residues were qualitatively veri- fied with a chlorine-specific microcoulometric detector. Gas-chromatographic liquid phases used for this study were coated on acid-washed and silanized support. Phases were 1,5 percent OV-17/1.95 percent QF-1 (column A); 4 percent SE-30/6 percent QF-1 (column B); and 3.5 percent QF-1/6.5 percent DC-200 (column C). Operating conditions were established to yield opti- mum separation with reasonable retention time for the latest eluting compounds. Experimental Design During the 12 months of 1968, household dust samples were collected from 28 households; blood samples were obtained from 28 heads of households and 27 spouses on the same sampling dates. Household dust samples consisted of the entire month's vacuum sweepings with at least two sweepings a week. Homes were selected on the following bases: location in Weld County, past co- operation with the program, pobability and consistency of continuing participation, and willingness of the fam- ily to participate in a long-term study requiring monthly sampling. The sampling group consisted of 16 urban control households, 4 farm households, and 8 house- holds with at least one member who was a pesticide formulator. Control participants had no known occupational expo- sure to pesticides and only minimal home-use exposure. Farm participants were selected from those with highest agricultural usage of pesticides among participants in the Colorado Community Study on Pesticides (former- ly under Food and Drug Administration. U.S. Depart- ment of Health, Education, and Welfare; currently un- der Technical Services Division, U,S. Environmental Protection Agency — Office of Pesticide Programs). The fornuilator group ranged from those employed in local pesticide formulating plants to professional appli- cators who mixed and loaded pesticides for commercial use. Results and Discussion Table 2. a summary of each study group and substrate, lists total number of samples, frequency of occurrence, mean of values above the minimtim sensitivity limits, and range of values for all pesticides detected during the I -year sampling period, Gas-chromatographic col- umn C was utilized to analyze all samples through June 1968 and did not provide adequate resolution or sensi- tivity for determining chlordane in blood. Samples an- alyzed after June were analyzed on gas-chromato- graphic columns A and B utilizing a modification of the florisil cokimn eltition of two fractions improving reso- lution of pesticides occurring in household dust samples. Because of widespread usage of DDT over the past 25 years, residues occtirring most frequently in blood were p.p'-DDT and its metabolite, p,p'-DDE; in household dust, the most common residue was p.p'-THyT. Also occurring frequently in household dust was o,p'-DDT, the technical isomer of p,p'-DDT. It did not occur in the blood samples because it is rapidly metabolized and excreted. Table 3 is a statistical evaluation of the 12- month average of p,p'-DDT and p,p'-DDE in blood, and p,p'-DDT in household dust. Correlation coefficients based on all 28 households showed a statistically significant positive correlation. It was observed, however, through inspection of data from each household that one household consistently yielded 210 Pesticides Monitoring Journal TABLE 2. Pesticide residues in human blood sera and household dust in 28 households, Weld County, Colo. — 1968 Control Group Farmer Formulator Control Group Farmer Formulator No. Samples M 187 F 171 M 46 F 47 M 88 F 87 182 45 95 Blood Residues, ppb HousEDusT Residues, ppm p,p'-DDT F R X 94 5-14 7.7 96 5-68 8.1 36 5-19 9.5 11 5-6 5.4 84 5-68 17.9 56 5-21 9.4 159 1.56-35.44 6.90 42 1.60-37.80 8.87 95 2.34-226.15 30.66 P,P'-DDE F R X 187 5-95 28.3 170 5-105 24.3 46 14-116 48.4 47 7-28 17.0 88 16-209 54.8 87 9-82 30.1 4 1.50-12.28 4.37 9 1.63-7.55 3.28 46 1.50-17.10 4.83 o,p'-DDT F R X 56 1.52-10.20 2.67 16 1.62-5.31 2.91 55 1.63-21.99 6.31 p.p'-DDD F R X 1 5 5.0 2 1.73-1.81 1.77 8 2.22-19.21 6.11 9 1.59-7.03 2.95 Methoxychlor F R X 78 1.53-28.57 6.09 21 1.58-102.90 14.93 56 1.92-144.44 18.24 Lindane F R X 11 " 7-23 16.8 1 5 5.0 3 1.75-2.27 2.05 43 1.54-13.72 5.85 g-BHC F R X 15 5-8 5.9 7 9-15 10.9 19 5-27 12.4 10 5-12 8.8 25 6-59 18.7 11 6-22 9.5 Chlordane F R X 6> 60-233 151 45 1.79-41.36 7.59 10 1.92-10.72 5.79 77 2.15-135.78 23.11 Dieldrin F X 1 5 5.0 47 5-140 43.5 13 = 5-14 10.1 25 1.59-10.21 2.94 15 1.59-10.63 4.42 59 1.59^0.42 8.92 Heptachlor Epoxide F R X 21 7-22 10.9 Endrin F £. X 27 1.50-27.73 7.14 Dacthal F R X 14 1.53-32.59 7.11 22 1.90-59.71 18.50 19 1.55-41.91 7.28 NOTE: F = Frequency of values above minimum sensitivity limit. R — Range of values above minimum sensitivity limit. X = Mean of values above minimum sensitivity limit. ' One participant only. 2 Eleven values from one participant. residue levels several magnitudes above the means of the other households. By omitting data from the one household, the correlations remained positive but lost any statistical significance, as demonstrated in Table 3. Hence the contribution of the one household to the sig- nificant correlation was disproportionate. Mean p,p'-DDT in the one household's dust was 131.14 ppm; in the other seven formulator households the mean was 16.13 ppm. Mean p,p'-DDT and p.p'-DDE in blood of the male occupant of the household with highest resi- dues were 46.8 ppb and 143.3 ppb, respectively, where- as male occupants of the other seven households had means of 12.3 ppb and 40.9 ppb, respectively. Although these data suggest a trend, they do not prove statistical correlations between p,p'-DDT levels in household dust and p, p' -DDT / p,p' -DDE levels in human blood. Interpretations of frequencies and ranges of some pesti- cides listed in Table 2 can be made. The relatively high frequency of chlordane occurrence in control household dust was caused by its use in many common household insecticides. Occurrence of the herbicide Dacthal in farm household dusts was caused by its use on certain produce croplands in the Weld County area. Dacthal occurred in lower levels among formulators and control groups, probably because these individuals had no history of occupational exposure to this pesti- cide. Three values requiring further explanation are chlordane and lindane in the blood of a male formulator and dieldrin in the blood of his wife. These unusual findings of chlordane and lindane in blood and a related medical problem in this male participant have been published previously (8). Mean chlordane and lindane levels in the dust from this home were 76.46 ppm and 10.90 ppm, respectively; in the other seven formulators' homes mean values were 10.18 ppm and 1.58 ppm. respec- VoL. 8, No. 3, December 1974 211 lively. Because neither chlordane nor lindane residues are commonly found in human blood except in acute exposure situations, this observation seems to present circumstantial evidence relating household dust residues to body burden. The mean dieldrin level in this subject's household dust was 28.02 ppm compared to 2.29 ppm for the other seven formulators' households. As a result. 11 of 13 occurrences of dieldrin in the blood of formu- lators' wives were obtained from this participant. This case provides further circumstantial evidence relating household residue levels to blood residue levels because this woman had no history of occupational exposure to dieldrin. TABLE 3. Correlation coefficients for levels of DDT in household dust with levels of DDT and DDE in human blood, Colorado— 1968 Household Dust ' P.P'-DDT 28 Households = 27 Households Blood P.p'-DDT P.P'-DDE 0.81 0.75 0.25 0.11 NOTE; Product-moment correlation coefficients. Coefficients represent 12-month average. ^ Coefficients first calculated for 28 households, then recalculated omit- ting one household with extremely high residues which overly influ- enced correlations; data could not be considered statistically signifi- cant. -Values significant at the 0.01 level. Conclusions Data from this study have shown no quantitative rela- tionships between pesticide levels in household dust and pesticide levels in blood. Circumstantial data from indi- vidual households indicate that at least some relation- ships do exist; however, no statistical significance can be demonstrated. It is evident from the current litera- ture as well as data presented here that a simplified ap- proach to the problem of relating pesticide body burden to a specific type of exposure, either environmental or dietary, is difficult because of the many variables in this complex problem. Acknowledgments Authors are indebted to Dr. Wayne D. Guenzi, Soil Conservation Service, U.S. Department of Agriculture, Fort Collins, Colo., for his suggestions in helping to develop the analytical method, and to the staff of the Colorado Community Studies on Pesticides for many of the analyses. We are particularly grateful to Dr. Strother Walker and Dr. Gary Zerbe. Department of Biometrics. University of Colorado Medical Center, Denver, Colo., for statistical analyses. LITERATURE CITED (/) Campbell, J. E., L. A. Richardson, and M. L. Schafer. 1965. Insecticide residues in the human diet. Arch.i. Environ. Health. 10:831-836. (2) Walker, S. 1967. Chairman, Biometrics Department,, University of Colorado Medical Center, Denver, Colo.-. Personal communication. (3) Colen, J. A/., and C. Pinkerton. 1966. Widespread! translocation of pesticides by air transport and rainout.; Organic Pesticides in the Environment. Advan. Chem.i Ser. No. 60. 163-175. {4) Johnson, R. E.. and R. I. Starr. 1967. Ultrasonic ex- traction of insecticides in soil. In Comparison of extrac- tion methods and solvent systems over three time in- tervals. J. Econ. Entomol. 60:1679-1682. (.";) Fahcy, J. E.. J. W. Butcher, and R. T. Murphy. 1965. Chlorinated hydrocarbon insecticide residues in soils of urban areas. Battle Creek, Michigan. J. Econ. Entomol. 58:1026-1027. (rt) Burchfield, H. P., D. E. Johnson, and E. E. Storrs. 1965. Extraction of soil. Guide to the Analysis of Pesticide Residues, Vol. 1, section II; A 2 (1); A 2a (1); A 2b (1) U.S. DHEW-PHS. (7) Dale, W . E., A. Ciirley, and C. Cueto. 1966. Hexane extractable chlorinated insecticides in human blood. Life Sci. 5:47-54. (S) Starr. H. G.. Jr.. and N. J. Clifford. 197 1. Absorption of pesticides in a chronic skin disease. Arch. Environ. Health. 22:396-400. 212 Pesticides Monitoring Journal BRIEF A Nomograph for the Conversion of 2,4-D Ester Concentrations in Air from ^g/nf to ppb^ and Vice Versa^ Raj Grover and Barry McCashin ABSTRACT A nomograph for the conversion of 2,4-D {2 ,4-Dichloro- phenoxyacelic acid) ester concentrations from ng/m^ to pph^. and vice versa lias been prepared to provide a simple and direct means for such conversions. Results obtained are shown to he comparable to those calculated. Introduction Data on direct measurements of pesticide residue levels in the atmosphere are being reported in increasing num- bers. Air pollution aspects (/) and hazards from off- target drift of agriculturally generated pollutants (2-4), especially pesticides, have recently been reviewed. Mea- surements of pesticide levels in the atmosphere are gen- erally expressed in weight/volume (w/v) units, i.e., nanogram or microgram of pesticide per cubic meter of air (ng or itg/m^). However, the cubic meter unit is dependent on ambient conditions of temperature and pressure which should be designated. When dealing with gaseous pollutants in the atmosphere, it is often desir- able to denote concentrations in relative volumes of w/v units, i.e., parts of pesticide per million or billion parts of air (ppm^. or ppb>). Because the ratio of volumes is unaffected by fluctuations in temperature and pres- sure during sampling, ppb^ or ppm, units do not change as volumes change. Procedures for conversion of concentrations from \igfmP to ppb^ or ppm^ have been discussed in recent texts on air sampling methodology (.5.6). However, nomographs provide a simple and direct means for such conversions and have been developed for a variety of gaseous compounds (7). ' Contribution from the Herbicide Behavior in Environment Section. Research Station. Agrictilturc Canada. Region. Saskatchewan. S4P 3A2. Canada. Concentrations of any gaseous compound in the air can be converted from (.ig/m^ to ppb,, using the following expression: |_ m3 J w X 10'' |_ ng J [_g mole vol J [parts "I billion parts J X 109 g mole vol = z [ppb,] where y = concentration of the residue in ug/m', w = gram molar weight of the compound, V = gram molar volume of air in m^, and z = concentration of residue in ppb. Example 1. Convert 40 jig/m^ methyl ester of 2,4-D sampled at 20°C and 1 atm pressure (STP) to parts per billion (ppb,.). 40 L ^' \ w = 235.08 (g mole wt) v = 0.024 r "• 1 [_ g mole vol J at STP Substituting the values for y, w, and v in the above ex- pression, 40 X 1 235.08 X 108 X 0.024 X 109 = 4.08 ppb^. Similar conversions for other 2,4-D esters were also carried out, using different temperature and pressure conditions (Table 1). In all these conversions, deter- mination of g mole volumes of air in m^ at various pressure and temperature conditions is necessary. While preparing the nomograph, extremes of pressure and temperature conditions most likely to occur in air Vol. 8, No. 3. December 1974 213 TABLE I. Relative conversion values for various 2,4-D esters, from ^g/m-' to ppb,, or vice versa, using calculations and the nomograph Sampling Conditions Conversion Values From To Pressure (MM Hg) Temi> (K) P/T Factor ' 2,4-D ESTER Calculation Nomograph methyl i50-propyl n-butyl /50-octyl 733 760 700 800 293 293 323 313 2.50 2.59 2.17 2.56 10 „g/m' 50 yg/m ■■' 8 ppb, 2 ppbv 1.1 ppbv 4.6 ppbv 77.1 ^e./m< 26.6 ^g/m ■' 1.1 ppbv 4.5 ppbv 77.0 ^g/m ■' 26.3 jig/m ■ 1 Pressure/temperature factor. sampling were selected. These were 20° to 50°C and 700 to 800 mm Hg. Pressure/temperature factors (P/T in mm Hg/K), in the range of 2 to 3, were then plotted against the ppb,. scale on the left side of the log-log paper (Fig. 1). Corresponding ester concen- trations were plotted on the right side of the log-log paper in units of i^ig/m^. A number of conversions similar to those calculated were read from the nomograph using the procedure shown in Figure 1. The two sets of values, i.e., those calculated and the ones obtained from the nomograph. showed close approximation (Table 1). Conversionsi beyond the range of the nomograph can also be carried out by simply shifting the decimal point to obtain the appropriate range. The resultant conversion value can then be adjusted to the correct value by placing the decimal point in the reverse direction. In conclusion, the nomograph greatly facilitates con^ version of atmospheric residue levels of 2,4-D esters from fig/m-' to ppb,. and vice versa. It provides a simple and direct means for such conversions while still main-i' taining a relatively high degree of accuracy. FIGURE 1. Nomograph for converting concentrations of four esters of 2,4-D from ng/m' to ppb,^ dry air at various pres sure/temperature factors (P/T in mm Hg/K) 214 Pesticides Monitoring Journai LITERATURE CITED /) Finkehteiii, H. 1969. Air pollution aspects of pesticides. Litton Systems Inc., Environ. Syst. Div., Bethesda, Md. 173 pp. 2) Grover, R.. J. May bank. K. Yo.shUta. and J. R. Ptim- mer. 1973. Droplet and volatility drift hazards from pesticide application. Preprint No. 73-106. 29 pp. J. Air Pollut. Contr. Ass., 66th Ann. Mtg., Chicago, 111. 3) Hay. J. R.. R. Grover. and K. S. McKinlay. 1971. Bio- logical significance of deposited pesticides. 59-64. In J. T. Bergsteinsson and W. Baier (Eds.) Meteorological Aspects of Pollution in Relation to Agricultural Pesti- cides. Canada Comm. Agr. Meteorol., Research Branch, Ottawa, Canada. (4) Heck. W. W., O. C. Taylor, and H. E. Heggestad. 1973. Herbaceous and ornamental plants and agriculturally generated pollutants. J. Air Pollut. Contr. Ass. 23:257- 266. (5) Anonymous. 1972. Methods of air sampling and analy- sis. Amer. Pub. Health Ass., Washington, D.C. 480 pp. (6) Ledbetter, J. O. 1972. Air pollution: Part A — Analysis. Marcel Dekker, Inc., New York. 424 pp. (7) Sheehy, J. P., W. C. Achinger, and R. A. Simon. 1968. Handbook of air pollution, U.S. Dept. of Health, Educ, and Welfare, Environ. Health Serv., Natl. Center for Air Poll. Cont., Durham, N.C. Vol. 8, No. 3, December 1974 215 APPENDIX Chemical Names of Compounds Discussed in This Issue ALDRIN BHC (Benzene hexachioride) CHLORDANE 2,4-D DCPA (Dacthal®) DDD DDE DDT DICHLOBENIL DIELDRIN ENDRIN FENAC HEPTACHLOR HEPTACHLOR EPOXIDE HEXACHLOROBENZENE (HCB) LINDANE METHOXYCHLOR POLYCHLORINATED BIPHENYLS (PCB's) TDE (DDD) Not less than 95% of I,2,3,4,10,10-Hexachloro-l,4.4a,5.8,8a-hexahydro-l,4-fndo-p.vo-5.8-dimethanonarhthaIene 1,2.3.4,5,6-HexachlorocycIohexane (mixture of isomers). Commercial product contains several isomers of which gammti is most active as an insecticide. l,2,3,5,6,7,8.8-Octachloro-2,3.3a,4,7,7a-hexahydro-4,7-methanoindene. The technical product is a mixture of several compounds, including heptachlor, chlordene, and two isomeric forms of chlordane. 2,4-Dichlorophenoxyacelic acid. Dimethyl 2,3,5,6-tetrachloroterephthalate See TDE. Dichlorodiphenyl dichloro-ethylene. (Degradation product of DDT.) Main component; l,l-Dichloro-2.2-bis(p-chlorophenyl) ethylene o,p'-DDE l.l-Dichloro-2-(o-chlorophenyl)-2-(/'-chlorophenyl ) ethylene p.p'-DDE l.l-Dichloro-2,2-bis(p-chlorophenyl ) ethylene -Bis (p-chlorophenyl) , , -trichloroethane. Numerous isomers in addition to p.p'-DDT are possible, and* some are present in the ctmimercial product. o,p'-DDT [I,l,l-Trichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl) ethane] 2,6-Dichlorobenzonitrile Not less than 85% of 1,2, 3,4, 10.10-Hexachloro-6.7-epoxy-l, 4.4a. 5. 6.7,8, 8a-octahydro-l,4-endo-fio-5.8,-dimethano- naphthalene l,2,3,4,10,10-Hexachloro-6,7-epoxy-l,4,4a,5,6,7,8,8a-octahydro-l,4-e«do-e'n(fo-5,8-dimethanonaphthalene 2,3,6-Trichlorophenylacetic acid l,4,5,6,7,8,8-Heptachloro-3a.4,7,7a-tetrahydro-4,7-endo-methanoindene 1.4.5,6,7,8,8-Heptachloro 2,3-epoxy-3a,4.7,7a-tetrahydro^.7-methanoindane Perchlorobenzene Gamma isomer of benzene hexachioride (1.2.3,4.5.6-hexachlorocyclohexane) of 99 + % purity 1 ,1 , l-Trichloro-2,2-bis (p-methoxyphenyl ) ethane Mixtures of chlorinated biphenyl compounds having various percentages of chlorine. 2.2-Bis (p-chlorophenyl)-l,l-dichloroethane (including isomers and dehydrochlorination products) 216 Pesticides Monitoring Journal Information for Contributors The Pesticides Monitoring Journal welcomes from all sources qualified data and interpretive information which contribute to the understanding and evaluation of pesticides and their residues in relation to man and his environment. The publication is distributed principally to scientists and technicians associated with pesticide monitoring, research, and other programs concerned with the fate of pesticides following their application. Additional circulation is maintained for persons with related in- terests, notably those in the agricultural, chemical manu- facturing, and food processing industries; medical and public health workers; and conservationists. Authors are responsible for the accuracy and validity of their data and interpretations, including tables, charts, and refer- ences. Accuracy, reliability, and limitations of the sam- pling and analytical methods employed must be clearly demonstrated through the use of appropriate procedures, such as recovery experiments at appropriate levels, confirmatory tests, internal standards, and inter-labora- tory checks. The procedure employed should be ref- erenced or outlined in brief form, and crucial points or modifications should be noted. Check or control samples should be employed where possible, and the sensitivity of the method should be given, particularly when very low levels of pesticides are being reported. Specific note should be made regarding correction of data for percent recoveries. Preparation of manuscripts should be in con- formance to the CBE Style Manual, 3d ed. Coun- cil of Biological Editors, Committee on Form and Style, American Institute of Biological Sciences, Washington, D. C, and/or the Style Manual of The United States Government Printing Office. An abstract (not to exceed 200 words) should accompany each manuscript submitted. All material should be submitted in duplicate (original and one carbon) and sent by first-class mail in flat form — not folded or rolled. Manuscripts should be typed on 8'/2 x 1 1 inch paper with generous margins on all sides, and each page should end with a completed paragraph. All copy, including tables and references, should be double spaced, and all pages should be num- bered. The first page of the manuscript must con- tain authors' full names listed under the title, with affiliations, and addresses footnoted below. Charts, illustrations, and tables, properly titled, should be appended at the end of the article with a notation in text to show where they should be inserted. -Charts should be drawn so the numbers and texts will be legible when considerably reduced for publication. All drawings should be done in black ink on plain white paper. -Photographs should be made on glossy paper. Details should be clear, but size is not important. -The "number system" should be used for litera- ture citations in the text. List references in the order in which they are cited in the text, giving name of author/ s/, year, full title of article, exact name of periodical, volume, and inclusive pages. The Journal also welcomes "brief" papers reporting monitoring data of a preliminary nature or studies of limited scope. A section entitled Briefs will be included, as necessary, to provide space for papers of this type to present timely and informative data. These papers must be limited in length to two journal pages (850 words) and should conform to the format for regular papers accepted by the Journal. Pesticides ordinarily should be identified by common or generic names approved by national scientific so- cieties. The first reference to a particular pesticide should be followed by the chemical or scientific name in parentheses — assigned in accordance with Chemical Abstracts nomenclature. Structural chemical formulas should be used when appropriate. Published data and information require prior approval by the Editorial Advisory Board; however, endorsement of published in- formation by any specific Federal agency is not intended or to be implied. Authors of accepted manuscripts will receive edited typescripts for approval before type is set. After publication, senior authors will be provided with 100 reprints. Manuscripts are received and reviewed with the under- standing that they previously have not been accepted for technical publication elsewhere. If a paper has been given or is intended for presentation at a meeting, or if a significant portion of its contents has been published or submitted for publication elsewhere, notations of such should be provided. Correspondence on editorial matters or circulation mat- ters relating to official subscriptions should be addressed to: Paul Fuschini, Editorial Manager, PESTICIDES MONITORING JOURNAL, Technical Services Divi- sion. Office of Pesticides Programs, U. S. Environmental Protection Agency, Room B49 East, Waterside Mall, 401 M Street, S.W., Washington, D. C. 20460. ■,:, U.S. GOVERNMENT PRINTING OFFICE: 1974-621-556/2 Pesticides Monitoring Journal The Pesticides Monitoring Journal is published quarterly under the auspices of the FEDERAL WORKING GROUP ON PEST MANAGEMENT (responsible to the Council on Environ- mental Quality) and its MONITORING PANEL as a source of information on pesticide levels relative to man and his environment. The WORKING GROUP is comprised of representatives of the U.S. Departments of Agricul- ture; Commerce; Defense; the Interior; Health, Education, and Welfare; State; Transportation; and Labor; and the U.S. Environmental Protection Agency. The pesticide MONITORING PANEL consists of representatives of the Agricultural Research Service, Animal and Plant Health Inspection Service, Extension Service, Forest Service, Depart- ment of Defense, Fish and Wildlife Service, Geological Survey, Food and Drug Administration, Environmental Protection Agency, National Marine Fisheries Service, National Science Founda- tion, and Tennessee Valley Authority. Publication of the Pesticides Monitoring Journal is carried out by the Technical Services Divi- sion, Office of Pesticides Programs of the Environmental Protection Agency. Pesticide monitoring activities of the Federal Government, particularly in those agencies repre- sented on the pesticide MONITORING PANEL which participate in operation of the national pesticides monitoring network, are expected to be the principal sources of data and interpretive articles. However, pertinent data in summarized form, together with interpretive discussions, are invited from both Federal and non-Federal sources, including those associated with State and community monitoring programs, universities, hospitals, and nongovernmental research institu- tions, both domestic and foreign. Results of studies in which monitoring data play a major or minor role or serve as support for research investigation also are welcome; however, the Journal is not intended as a primary medium for the publication of basic research. Manuscripts received for publication are reviewed by an Editorial Advisory Board established by the MONITORING PANEL. Authors are given the benefit of review comments prior to publication. Editorial Advisory Board members are: John R. Wessel, Food and Drug Adminislration, Chairman Paul F. Sand, Agricultural Research Service, Vice Chairman Anne R. Yobs, Center for Disease Control William F. Durham, Environmental Protection Agency Thomas W. Duke, Environmental Protection Agency G. Bruce Wiersma, Environmental Protection Agency William H. Stickel, Fish and Wildlife Service Milton S. Schechter, Agricultural Research Service Herman R. Feltz, Geological Survey Mention of trade names or commercial sources in the Pesticides Monitoring Journal is for identification only and does not represent endorsement by any Federal agency. Address correspondence to: Paul Fuschini Editorial Manager PESTICIDES MONITORING JOURNAL U.S. Environmental Protection Agency Room B49 East, Waterside Mall 401 M Street, S.W. Washington, D. C. 20460 Martha Finan Joanne Sanders Editors CONTENTS Volume 8 March 1975 Number 4 PESTICIDES IN PEOPLE Page Blood organochlorine pesticide levels in Virginia residents 219 Francis D. Griffith, Jr., and Robert V. Blanke PESTICIDES IN WATER Fate of copper in ponds 225 A. W. Mcintosh RESIDUES IN FOOD AND FEED Preliminary survey of ethylenethioiirea residues in the Canadian food supply 232 Zigmund Pecka, Patricia Baulu, and Harvey Newsome RESIDUES IN FISH, WILDLIFE, AND ESTUARIES Pesticide and mercury residues in commercially grown catfish 235 A. B. Crockett, G. B. Wiersma, H. Tai, and W. Mitchell Residues of methoxychlor and other chlorinated hydrocarbons in water, sand, and selected fauna following injections of methoxychlor black fly larvicide into the Saskatchewan River, 1972 241 F. J. H. Fredeen, J. G. Saha, and M. H. Balba Organochlorine residues in starlings, 1972 247 Paul R. Nickerson and Kyle R. Barbehenn Organochlorine residues in Alaskan peregrines 255 David B. Peakall. Tom J. Cade, Clayton M. White, and John R. Haugh GENERAL Comparison between two methods of subsampling blubber of northern fur seals for total DDT plus PCB's 261 Raymond E. Anas and Donald D. Worliind Degradation of parathion applied to peach leaves 263 Wray Winterlin, J. Blair Bailey, Larry Langbehn, and Charles Mourer APPENDIX Chemical names of compounds discussed in this issue 270 ERRATUM 271 ACKNOWLEDGMENT 271 ANNUAL INDEX (VOLUME 8, JUNE 1974— MARCH 1975) Preface ^ 272 Subject index 273 Author index 278 Information for contributors 280 ^/^^t^ PESTICIDES IN PEOPLE Blood Organochlorine Pesticide Levels in Virginia Residents Francis D. Griffith, Jr.,i and Robert V. Blanke 2 ABSTRACT This study attempts to establish 1972 baseline levels for 31 organochlorine pesticides anil industrial chemicals in post- mortem human whole blood in Virginia. These pesticides and industrial chemicals have been detected previously in other parts of the food chain and environment. In the present study DDT and its metabolites, DDE and TDE, were detected most frequently and at highest levels. DDT and DDE tended to appear more frequently as people grew older although TDE levels remained constant. Diel- drin and lindane showed peak levels in the middle age group. Analyzing distribution of pesticides in blood by sex showed that females had higher levels of lindane and dieldrin and rtiales had higher levels of DDT, DDE, and TDE. Analyzing racial distribution showed blacks with higher levels of DDT, TDE, and DDE and indicated little difference from whites for lindane and dieldrin. Higher levels were found in Rich- mond and Norfolk than in the Fairfax and Roanoke re- gions. Introduction This study attempts to establish 1972 baseline levels of organochlorine pesticides in human whole blood in Virginia. It follows by 6 to 12 months the completion of a basic study by the Virginia Department of Agri- culture and Commerce (/) required by the Virginia General Assembly (2) on the occurrence of economic poisons in the environment. Until now, little human monitoring has been done nationally and no studies have been performed on the population of Virginia. Pesticides which have been detected in the lower part of the food chain and the environment are those most 1 Division of Consolidated Laboratory Services, Commonwealth of Virginia, Richmond, Va. 23219. = Department of Pathology, Medical College of Virginia, Richmond, Va. likely to occur in human blood. The study tested for hard organochlorine pesticides as well as some of their alternatives, such as metho.xychlor and endosulfan, and some fungicides, such as captan. Polychlorinated bi- phenyls (PCB's), manufactured as industrial chemicals since the late 1920's, have worked their way through the environment to become manifest in eggs, poultry, fish, beef, packed cereal, and water. Another reason for monitoring PCB's is that several estuaries in Virginia have been contaminated with heavy industrial dis- charges. Postmortem blood was selected as the specimen to be examined for the following reasons : Collection techniques and consent requirements from living patients residing in geographically dif- ferent regions of the State required facilities be- yond authors' control. Future monitoring can most conveniently be per- formed on postmortem material; the existing state- wide Medical Examiner's system provides a ready mechanism for specimen collection. Four centers throughout Virginia are assigned to collect specimens from subjects varying widely in age, sex, race, residence, and occupation and make retrievable computer records available on each case. Similar studies in other areas have used post- mortem material (5). The disadvantage of postmortem blood is that it differs from blood of living patients. Some decomposition has occurred, clotting factors are altered, intracellular and extracellular distributio/i of diffusible substances are altered, and the ratio of formed elements to serum may be modified. Vol. 8, No. 4, March 1975 219 Sampling Procedures Blood samples were taken by pathologists in the four regions of the Office of the Chief Medical Examiner: Richmond. Roanoke, Norfolk, and Fairfax. The choice of samples for analysis was based on availability. Vari- ables such as age. race. sex. occupation, residence, and cause of death were not considered initially. Deaths had resulted from trauma, violence, or suspicious, unusual, or unnatural causes; during imprisonment; without med- ical attendance; suddenly, to a victim who had been in apparent good health; or following surgical or anes- thetic procedures (4). Pathologists were requested to take blood within 24 hours after death, although samples drawn as late as 72 hours after death were accepted. Samples were drawn from the heart and placed in 15-cc tubes containing 120 mg sodium fluoride and 45 mg potassium oxalate. To the tube was attached a label bearing the name and address of the deceased, date of death, and the Medical Examiner's signature. In Richmond the tubes were sealed and refrigerated until analysis. In the other regional offices, the tubes were sealed, mailed to Richmond in cardboard containers, and refrigerated upon arrival. After analysis samples were frozen at —25° C for fu- ture use. A nalytical Procedures Aldrin and dieldrin have been extensively used through- out Virginia in recent years. Thus analytical procedures, especially the extraction step, were keyed to the recov- ery of dieldrin (5). which has been detected more fre- quently in the food chain than has DDT and is five times more toxic to rats and mice on an LDr,,, basis (6). EXTRACTION The whole-blood extraction procedure was a modifica- tion of the sulfuric acid method of Henderson. De Boer, and Stahr used in the 1972 Association of Official An- alytical Chemists (AOAC) collaborative study for mul- tiresidues in whole blood (7). This method was chosen because sulfuric acid released bound pesticides and sul- fonated and removed some of the unsaturated interfer- ences (8). Two ml whole blood was pipetted into a 50-ml centrifuge tube; 1.5 ml 60 percent H^SO, was added and mixed on a vortex mixer for 5 seconds; a second 1.5 ml 60 percent H^SO, was added and mixed for 10 seconds; and 2 ml 60 percent H^S04 was added and mixed for 30 seconds. Samples were then cooled for several minutes. Pesticides were extracted with 3 ml 9:1 pesticide-grade hexane:acetone. Samples were mixed 20-30 seconds on a vortex mixer and then centrifuged at 2000 rpm for 10 minutes. The hexane-acetone layer was removed with a disposable capillary pipette and the pesticide solvent layer was put into a 1 3-ml Kontes graduated centrifuge tube. Hexane-acetone extraction was repeated twice more. All three solvent layer extrac- tions were combined in the same tube. The solvent was concentrated to 0.5 ml by using a gentle stream of clean dry nitrogen on the surface of the extract. ANALYSIS Gas-liquid chromatographic (GLC) determinations were performed as described by Griffith and Blanke at the 87th AOAC Annual Meeting (9). Samples were run on a Dohrmann Model 2468 gas chromatograph with a microcoulometric gas titrating system (GTS-20) for halogens. Confirmations were made on a Micro-Tek GC-2000-R gas chromatograph equipped with a tritium foil electron-capture detector. The column used in the microcoulometric system was a 6-ft-by-4-mm-ID glass column packed with 5 percent OV-210 on 80/100 mesh Gas-Chrom Q. The tempera- ture was programmed from 210° to 234° Cat 2° C/min. The column used in the electron-capture system was a 6-ft-by-4-mm-ID glass column packed with 4 percent SE-30/6 percent QF-1 on 80/100 mesh Supelcoport. The temperature was isothermal at 205° C. Carrier gas for both systems was nitrogen. Flow through the OV- 210 column was 90 ml/min.; flow through the SE-30/ QF-1 column was 120 ml/min. Qualitative results were based on the relative retention of aldrin to the retention of the pesticide in question. Quantitation of pesticides was based on peak area using a disc integrator. Recovery data were obtained for all compounds dis- cussed. Most analyses were single determinations be- cause precision experiments showed no improvement with triplicate analyses. The desired minimum detect- able recovery for all pesticides was 1 ppb. Alpha BHC. lindane, heptachlor. aldrin. heptachlor epoxide. CIPC, and dieldrin were recovered at the 1 ppb level; the limit of detection for DDE, DDT, TDE, dacthal, en- drin, endosulfan, and atrazine was 2 ppb. Recoveries for chlordane. dicofol. folpet. captan, chloropropylate. PCNB, carbophenothion. phosphamidon. methoxychlor, and toxaphene varied between 10 and 40 ppb. The min- imum detectable amount for PCB's (Aroclor 1221. 1232. and 1242) and PCN's (Halowax 1099) was 100 ppb because of interference from the background of the blood extract. PCB's (Aroclor 1254 and 1260) and PCN (Halowax 1014) had later eluting fractions that did not overlap the backgrounds of the blood extracts; hence the minimum detectable amounts for these chemi- cals was 50 ppb. Results and Discussion When all cases examined in this study were grouped by age (Fig. 1 ) a normal distribution was apparent, with the majority of samples in the 41- to 60-year age group. The mean (x) age was 45.98 years. The age distribution in the Richmond and Fairfax regions followed this pat- 220 Pesticides Monitoring Journal tern but distribution showed a majority in the 21- to 40-year age group in the Norfolk and Roanoke regions. The mean age of Virginia residents in 1972 was 26.8 years; the mean age at death was 68.0 years. The Nor- folk region showed a slightly younger living population of 24.2 years. Richmond and Roanoke residents aver- aged 28.3 and 30.2 years, respectively. The mean age of Fairfax residents and the mean age at death of resi- dents of the other regions are unknown {10). Thus the population included in this study represents an older group than the average living Virginia resident but a younger group at death than that which occurs nor- mally. Se.x and race distributions also deviated from the aver- age population (Fig. 2). The racial distribution of the population examined showed a ratio of three whites to two blacks. In the Richmond and Norfolk regions, the racial distribution was 1:1. but whites represented a majority of those samples from the Fairfax and Roan- oke regions. The male: female distribution was about 3:1 in all four regions of the State. The male: female ratio throughout Virginia is about 1:1 and the average white: black distribution is 4:1. Thus data from the present study represent a greater propor- tion of males and blacks than is typical of the State's population {10). If all data from this study are aver- aged, the resulting baseline of pesticide levels would be misleading. However, when results in each group are taken on the basis of age, sex, race, and geographical residence, they should be comparable to those in cor- responding living groups. Similar techniques are com- monly used to evaluate data derived from mortality tables (//). On a statewide basis, I 5 pesticides and industrial chem- icals were positively identified. Ten of these are shown in Figure 3. The remaining five did not occur in enough samples to be included. The most frequently detected pesticides were DDT and its metabolites, dieldrin. lin- dane, and alpha BHC. Less frequently detected were methoxychlor, heptachlor epoxide, CIPC, and PCB's. Captan was detected at levels of 20 to 30 ppb. and carbophenothion was found at levels of 8 to 60 ppb in two samples each, Chloropropylate. PCNB, and en- "■49' 40 214 20- %ot 142 %of,5 74 95 ^°^*^ y= in. 36 ZO 1 0-1'^ ,v.*^ av-^ 6^-60 W 0-lP ZV'*° >,^-^ 6^-«° 6^' 1 AGE. YEARS AGE, VEARS 1 NORFOLK , FAIRFAX , ROArWKE | n=e9 r-76 n=40 10 41 '° 44 10 33 %ot %ot 5 19 %o( 5 TOTAL TOTAL ^ TOTAL ^ '^ 13 _fl^ 6 1 5 8 Ho ^nVio 0 '^t,-»°..-*6>-«° 8'" [a-'°.>-^6-*°«^- K> '°..-***v-«^ •^• AGE. YEARS AGE, YEARS AGE, YEARS 24 20 t le: i t 12 8 4 1 1 116 H'-A^, 7 n PI Mean. Poiilive Sample n Mean, All Samples H Kl >.•.. L c« ^.•-c )•• r a "' •1 1 1 UNMOfOUIIVU j-iB J- HI j-m m-m ,.„ LD-a '.« 1-1' FIGURE 1. ^,£?e distribution of humans sampled for pesticide residues FIGURE 3. Organochlorine residues detected in human blood STATE TOTAL RICHMOND 50 ■ 221 "-'S' 25- n = 292 liL, 105 if. %ot 30 TOTAL '5 TOTAL 71 41 35 10 n n ^] 1 1 ■ULf nttUS HALE FEMALf HALE FEMALE MALE 1 NORFOLK , FAIRFAX ROANOK E 10- n = 89 10 p-| n:76 10- n=40 J ^ M H %ot ^ 27 TC TAI t. TOTALS-' 10 n h nh 2 6 rn 7 n 0 FIGURE 2. Sex and race distribution of humans sampled for pesticide residues drin were each detected in one sample at 27, 1 5, and 5 ppb, respectively. Of equal importance are the 16 pesticides and industrial chemicals that were not detected. Chlordane was not detected; nor were the herbicides atrazine and dacthal, probably because of their rapid degradation. Toxa- phene, the PCN's, and most PCB's did not appear, Heptachlor and aldrin were not found, although their metabolites dieldrin and heptachlor epoxide were. The latter occurred in only 1 .4 percent of the samples stud- ied at an average concentration of 0.06 ppb. This is a significant decrease from the mid-sixties when that metabolite was first detected in milk {12). The current findings may indicate that milk is no longer a significant Vol. 8, No. 4, March 1975 221 source of PCB contamination, although such a conclu- sion could be verified only through extensive research into several areas, including the dietary habits of sub- jects whose blood was sampled. Methoxychlor. CIPC, and PCB's were detected in a small percentage of samples with wide difTerences be- tween frequency and concentration means (Fig. 3). Authors interpret this to indicate that they do not have widespread distribution and, when detected, probably indicate recent exposure. Concentrations and frequencies of DDE and DDT in blood by age showed similar patterns (Fig. 4,5). On a statewide basis, higher concentrations and occurrences were noted with advanced age. On a regional basis. Richmond and Norfolk had generally higher DDE and DDT concentrations and occurrences than had Fairfax and Roanoke. Distribution of TDE by age differed from DDT and DDE: it was fairly uniform except in the Norfolk region (Fig. 6). Average concentrations were higher in the 0- to 20-year age group although percent occurrence was lower in this group. Dieldrin and lindane occurred less frequently but gen- erally showed peak levels at middle age (Fig. 7.8). In the case of lindane, no positive cases were found among the very old and only one was detected in the very young. Dieldrin distribution was skewed to an earlier age group in the Roanoke and Norfolk regions than in the other two. In Norfolk, dieldrin levels were generally lower except in the very young. Distribution of detected pesticides by sex and race did not show any remarkable trends (Fig. 9). Females had higher levels of lindane and dieldrin; males had higher levels of DDT. DDE. and TDE. Blacks had higher levels of DDT and its metabolites but displayed little difl'erence from whites in lindane and dieldrin concen- trations. Higher pesticide levels and greater percent occurrences were found in Richmond and Norfolk than in Fairfax and Roanoke. No correlations between residue levels and occupation could be considered because of the wide variation in oc- cupations listed on Medical Examiner's certificates. Many people over 60 were listed as retired; others were unemployed. Although some pesticides are known to alter micro- somal enzyme activity and, consequently, dmg metab- olism (13). the low number of deaths by barbiturate overdosage prohibited observation of any relationship between pesticide levels and drug levels. In no case was pesticide poisoning considered the cause of death. Had it been, the pesticide would have been present in much higher levels, as observed in published findings on known overdosages and exposures (14). FIGURE 4. DDE residues delected in human blood by age distribution 20 le 16 1 « 1 '2 t 10 8 6 4 2 1 in 1 1 JIflL 1 1 1 1 HMOKD 1 1 i ,. 1 Mean. t^iiiiH Sainp4ei O Mean, All Sample! ■ « 1 ROANOKE .„ ..i.» „ . ...„ t-. ,.., ...,„- „,l.,. .,. FIGURE 5. DDT residues detected in human blood t>y a^e distribution 20 18 «:: a 12 ^ 10 8 6 4 2 s 1 T»TE TOTAL 1 ON 1 1 N 1 .. ' Mean. P AIRFAX n. 1 dmpiei ■ RMNOKE 1 1 1 1 ■ i - 1 1 ■ -| \ . < I> > 1 •.u vt> ).•> x'l )■<• )->* - !■■> ,. u - - >.^ t-» i-ti FIGURE 6. TDE residues detected in human blood b\ (liie distribution 222 Pesticides Monitoring Journal FIGURE 7. Diclclrin residues detected in human blood by age distribution Ntean, PosHife Sampfei C wwn, All^efnpKi ■ 7 STlTt TOTAL RCHPiOND NORFOLK FAIRFAX ROANOKE 6 1' - r — 1 2 3 2 r _ . ■ ■J ■ J 1 I " 1 1 'U • •' °-n !'-• !>■ ..-« ti- EkRi .,.- •.. I^-B.I-« • 1' - U-.C. . >.• ,.„ .« ... ». . V. . ..« >->: - ,-t - FIGURE 8. Lindane residues detected in human blood by age distribution Conclusions Pesticide levels in human blood of Virginia residents generally coincide with levels found in other studies. Exceptions probably stem from improvements and re- finements in analytical methods, differences in time peri- ods during which subjects were exposed to environ- mental pollutants, and variations in sample populations. Levels of DDT and its metabolites are very similar to those reported by Dale et al. in Georgia (15), slightly higher than those obtained by Davis and Edmundson et al. in Florida (16,17) and by Watson et al. in Idaho (18), and lower than results reported by Keil et al. in South Carolina (19). The percent occurrence of DDT and its metabolites found in this study is much lower than that reported by Watson et al. (18). This may parallel the decline of DDT residues in lower parts of the food chain in Virginia which was first noticed in 1970. The slightly higher occurrence of dieldrin in samples from the central region of Virginia coincides with the frequency with which this pesticide is detected in the food chain. Most samples of water from the James River basin contain low levels of dieldrin. Lin- dane occurred more frequently than had been antici- pated from previously reported pesticide residue analy- ses (1,20). This may result from the common use of lindane in commercial vaporizers and its presence in cigarette smoke (21). Differences in residue levels and percent occurrence by sex were not striking. Racial differences observed for all pesticides can be accounted for by various socio- economic factors discussed by Davis et al. (16). Residue levels detected did not indicate a health hazard from environmental contamination by pesticides in Vir- ginia communities (22). Although none of the differ- ences are statistically significant when applying the chi-square test, hopefully the data acquired will estab- lish a baseline for residents of this State. Continuous monitoring should detect significant changes. DDT levels are expected to drop but residues of DDT metab- olites may remain relatively constant or decrease at a lower rate. Dieldrin concentrations will probably re- main relatively constant or decrease at a lower rate than DDT. Future studies should be expanded to include additional pesticides and specimens such as adipose tissue and liver to better define overall pesticides body burden. It would also be useful to compare results in this study with a limited number of blood specimens from living patients of similar age, sex, race, and geographic origin to determine the suitability of postmortem blood as a sample for monitoring pesticide residues in humankind. FIGURE 9. Pesticide residues detected in human blood by sex and race distribution Vol. 8, No. 4, March 1975 223 LITERATURE CITED (/) Rone, M. B. {Commissioner). 1971. The 1971 Virginia pesticides study pursuant to house joint resolution 51. Virginia Department of Agriculture and Commerce, Richmond, Va. (2) DuVul, Van Clief, el at. (Patrons). Offered February 6, 1970. House joint resolution no. 51. Directing Dept. of Agriculture and Commerce to conduct a study on the need for regulation and control of economic poisons, Virginia General Assembly, Richmond, Va. (.?) Davies, J. £., W. F. Edmundson, M. J. Schneider, and J. C. Cassady. 1968. Problems of prevalance of pesti- cide residues in humans. Pestic. Monit. J. 2(2):8(l-85. (4) Medical E.xaminer's Law. Code of Virginia, Sec. 19; 1-41. (5) Saha, J. G., B. Bhavaraju, and Y. W. Lee. 1969. Validity of using soil fortification with dieldrin to measure solvent extraction efficiency. J. Agr. Food Chem. 17(4):874-876. (6) Sunshine, J. (ed.). 1969. Handbook of Analytical Toxi- cology. The Chemical Rubber Co., Cleveland, Ohio. Pp. 507 and 511. (7) Stretz, P. E., and H. M. Stahr. 1972. Collaborative study — determination of chlorinated pesticides in whole blood. Presented at 86th Annual Meeting of the Association of Official Analytical Chemists. Wash- ington, D. C. (8) Henderson. S. J., J. G. DeBoer, and H. M. Slahr. 1971. Improved method for determination of chlori- nated hydrocarbon pesticide residues in whole blood. Anal. Chem. 43(3) :445-447. (9) Griffith, F. D., Jr., and R. V. Blankc. 1974. Micro- coulometric determination of organochlorine pesticides in human blood. J. Ass. Offic. Anal. Chem. 57(3): 595-603. (10) Derr, B. P. 1972. Personal communication. Virginia Department of Health. Bureau of Vital Records and Health .Statistics, Richmond, Va. (//) National Center for Health Statistics. 1972. Vital Sta- tistics of the United States — 1968. Vol. II— Mortality. U.S. Department of Health, Education, and Welfare, Public Health Service, Health Services and Mental Health Administration, Washington, D. C. (12) Midyette, J. W . (Director). 1965. 1964-65 Annual Report. Commonwealth of Virginia, Department of Agriculture, Division of Technical Services, Rich- mond, Va. (13) Goth, Andres. 1970. Medical Pliarmacology. The C. V. Mosby Company, Saint Louis, Mo. Pp. 24-33. (14) Schafer, M. L. 1968. Pesticides in blood. Residue Rev. 24:19-39. (15) Dale, W. £., A. Ciirley, and W . ]. Hayes, Jr. 1967. Determination of chlorinated insecticides in human blood. Ind. Med. Surg. 36(4) :275-280. (16) Davis, J. £., W. F. Edmundson, D. Maceo, A. Bar- que t, and J. Cassady. 1969. An epidermiologic appli- cation of the study of DDE levels in whole blood. Amer. J. Pub. Health 59( 3) :435-441. (17) Edmundson, W. F., J. E. Davis, G. A. Nachman, and P. L. Roeth. 1969. F,p'-DDE in blood samples of oc- cupationally exposed workers. Pub. Health Rep. 84 (l):53-58. (18) Watson, A/., W . W . Benson, and J. Gabica. 1970. Serum organochlorine pesticide levels in people in southern Idaho. Pestic. Monit. J. 4(2):47-50. ( 19) Keil, J. £.. W. Weston III. C. B. Loadholi, S. H. Sandi- fer, and J. J. Colcobougli. 1972. DDT and DDE resi- dues in blood from children, South Carolina — 1970. Pestic. Monit. J. 6(1); 1-3. (20) Rowe, M. B. (Commissioner). 1971. Economic poisons and ecology first annual report 1970-71. Virginia De- partment of Agriculture and Commerce, Richmond, Va. (21) Schmid, K., and A. Rastetter. 1970. Gas chromato- graphic determination of insecticide residues in dried and fermented tobacco samples from field and growth experiments. Beitr. Tabakforsch 5(5 ) :201-206. (22) Hayes. W. J., Jr., W. E. Dale, and C. I. Pirkle. 1971. Evidence of safety of long-term high oral doses of DDT for man. Arch. Environ. Health 22:119-135. 224 Pesticides Monitoring Journai PESTICIDES IN WATER Fate of Copper in Ponds ^ A. W. Mcintosh 2 ABSTRACT Treatmenis of 3 ppm copper sulfate iCuSOi'^H,0) were applied to two small aquatic systems in Michigan in 197 1. To study the pathways of the added copper, samples of water, sediment, aquatic macrophytes, filamentous algae, and fish were collected and analyzed by atomic absorption. Sampling was initiated before treatment and continued up to 4 months in one of the ponds. Dissolved copper concentrations in water decreased rapidly immediately after treatment and then gradually to hack- ground levels. Reduction of dissolved copper may have involved initial precipitation of an insoluble compound, such as malachite, followed by sediment adsorption of soluble copper complexes and copper released from aquatic plants. Levels of copper in sediment increased rapidly at first and gradually later in the study. Aquatic plants and filamentous algae accumulated very high levels of copper. Uptake rates were apparently affected by water temperature and growth stages of the plants. Data indicate that aquatic macrophytes developing in one pond 10 weeks after treatment took up copper from the sediment. Although green siinfish (Lepomis cyanellus) accumulated copper soon after treatment, levels returned to background later in the study. Introduction Levels of copper generally below 20 ppb are routinely observed in natural waters. When higher values are found in surface waters, it is likely that the metal has been added by copper and brass tubing, industrial efflu- ents, or copper compounds used for control of unde- sirable aquatic organisms or plants (/). ' Deparlment of Fisheries and Wildlife. Michigan Slate University. East I ansiny. Mich. Research supported by Predoctoral Research Fellowship No. 5-Fl-WP-26.4.1h-02. Water Quality Office. U.S. En- vironmental Protection Agency, Washington. D.C. ^ Department of Bionucleonics, Purdue University, West Lafayette, Ind. 47906. Reprints available from this address. Limited research has been conducted concerning the fate of copper in aquatic systems, including copper pathways in canals and irrigation ditches (2-4), bogs (5), ponds (6,7), and lakes (8). In assessing the eco- logical effects of a heavy metal, components concentrat- ing it must be identified and characterized. Little effort has been devoted to determining the role of these aquat- ic system components in copper dispersal. In the present study in which copper sulfate (CuSOj-SHoO) was added to two small ponds in Michi- gan in 1971, intensive sampling of water, sediment, al- gae, plants, and fish was conducted for periods of 7 weeks to 4 months to assess the role of each component in copper distribution. Methods and Materials STUDY SITE A circular settling pond about 13.7 m in diameter and 2 m deep served as the study site. Located at the old East Lansing Sewage Treatment Plant, the pond is among a number of sites used by the Michigan State University Department of Fisheries and Wildlife for limnological research. In June 1970 the pond was drained and the bottom was covered with 10 to 15 cm black soil. The system was refilled and left undisturbed while populations of macrophytes and filamentous algae developed. Early in May 1971 the system was divided with a poly- ethylene sheet, effectively forming two separate ponds. By June, 100 green sunfish {Lepomis cyanellus) had been collected from other sources. They were weighed, marked by fin-clipping, and released into each system. Copper sulfate solutions were prepared in distilled water from commercial grade copper sulfate and applied by hand to the surface of each pond. Application dates and dosages appear in Table 1. Vol. 8, No. 4, March 1975 225 TABLE 1. Copper sulfate dosage rales and schedules Site Application Date Dosage, ppm Pond A Pond B July 12. 1971 September 26, 1971 3 3 NOTE: Ponds A and B are halves of one settling pond. Water chemistry parameters including pH, alkalinity, hardness, and dissolved oxygen were determined throughout the study (Fig. 1,2). Techniques outlined in Standard Methods for the Examination of Water and Wastewater (9) were used for chemical tests. Duplicate water samples were collected for analysis from the top and bottom quarters of each pond with a Kemmerer water sampler. r boliom o> pond FIGURE 1. Changes in water chemistry parameters. Pond A NOTE: Each point represents average of two samples. Shaded area denotes treatment with 3 ppm copper sulfate. Sampling and Analysis Water for copper analyses was collected at three stations established near the outside, middle, and inside of each semicircular pond. A water column was removed with a polyethylene tube and a subsample was collected. Preparation followed procedures outlined in Methods for Chemical Analysis of Water and Wastes (10). Cop- per passing through a 0.45-u membrane filter repre- sented dissolved copper; the fraction remaining on the filter was considered suspended copper. Samples When near top ol pond Samples taken neat bottom of pond 6-11 7-1 7-11 e-10 a- 30 9-1 DATE FIGURE 2. Changes in water chemistry parameters. Pond B NOTE: Each point represents average of two samples. Shaded area denotes treatment with 3 ppm copper sulfate. At each sampling period, two sediment samples were collected with a core sampler and frozen in dry ice. The top 1- 2-in. layer was removed and dried at 100° C for 96 hours. Samples were ground and 5 ml 8N HCI/g sediment was added. This slurry was held at a tem- perature near boiling for 20 hours, cooled, filtered, and diluted to volume with deionized distilled water. Plant samples were collected, rinsed in distilled water, blotted, and dried at 100° C for 96 hours. Tissues were digested by adding 10 ml 8N HNO^ and boiling to dryness. Five ml 6N HCl was then added and the mix- ture was reheated to dissolve the residue. The solution was filtered and diluted to the desired volume. Fish were trapped, sacrificed, rinsed in distilled water, weighed, and digested in 5 ml concentrated HNO;i/g fish for 24 hours. The solution was filtered and diluted to volume. 226 Pesticides Monitoring Journal Samples of all components were taken before treatment to determine background levels of copper (Table 2). Samples were collected after treatment for 4 months in Pond A and for 7 weeks in Pond B. TABLE 2. Background topper levels in two Michigan ponds, 1971 Average No. Samples Component Concentration, Copper, ppm Range, ppm WATER Dissolved copper ' 0.0145 0.00-0.05 38 Suspended copper - 0.0.157 0.00-0.09 38 Total 0.0502 0.00-0. n 38 SEDIMENTS ^ 27.68 1.08-73.60 51 PLANT TISSUE ^ Chara sp. 15.92 6.09-36.70 20 Oedogoniiim sp. .14.40 12.54-51.68 19 Elodea Ntillallii 26.31 2.33-50.51 28 Potamogeton crispiis 26.31 4.69-55.80 20 FISH« Green sunfish 0.56 0.00-1,64 22 1 Filtered through 45-^ membrane filter. - Fraction left on membrane filter. 2 Dry-weight basis. ^ Whole-body wet-weight basis. The presence of copper was determined with a Jarrell- Ash atomic absorption spectrophotometer Model 800. Sample concentrations were determined by comparing their readings to those of copper sulfate standards pre- pared several times during the tests. To insure accuracy, three standards were compared to those of another investigator; the discrepancy was less than 10 percent. During analyses, uncontaminated samples were spiked with copper to determine percent recovery. Recoveries for plant tissues ranged from 89.9 to 1 14.4 percent with a mean of 101.2 percent; recoveries from sediments ranged from 94.7 to 120.0 percent with a mean of 109.9 percent. Reproducibility was determined by reading the same samples on different occasions. When tested with a paired-t test, pairs of readings were not significantly different at the 5 percent level. Copper added to the ponds accumulated on upper sedi- ment surfaces. Because little downward movement of copper into sediment was expected during the test pe- riod, sediment copper values were expressed as weight accumulated on sediment surfaces. Average pretreat- ment copper concentrations were calculated in ^ig/g for pond sediment, and total |,ig copper present in the upper layer was determined for each core sample taken after treatment. The average jig/g pretreatment copper con- centration was subtracted from the total |^ig copper present in the upper layer after treatment to yield |^ig copper deposited on the upper surface of each core. Results and Discussion WATER Dissolved copper concentrations in Pond A diminished gradually after rapid decline of initial peaks (Fig. 3). High concentrations at stations Al and A2 shortly after treatment were probably caused by slow dispersal of copper sulfate. Malachite (Ciu(0H).jC03) precipita- tion, a slow process reaching equilibrium only several days after application ( / / ) . may have played a large role in the decline. ■ Dissolved copper concentrations -..•Suspended copper concentrations FIGURE 3. Changes in dissolved and suspended copper concentrations. Pond A, three sites NOTE: Shaded area denotes treatment with 3 ppm copper sulfate. Levels of organic acids in the range of 4 to 40 ppm carbon in water may increase the amount of metal stabilized in solution by several orders of magnitude {12). Ten sediment samples from Pond A contained an average of 1 1 percent organic matter; organic complexes in the overlying water may have slowed the decline of dissolved copper. Suspended copper levels in Pond A showed no trends. Although an initial peak occurred, concentrations of suspended copper soon declined to background levels (Table 2). Vol. 8, No. 4, March 1975 227 Dissolved copper concentrations in Pond B followed a pattern similar to that of Pond A following treatment (Fig. 4). Copper levels peaked initially, and subsequent- ly decreased. However, sudden increases in dissolved copper occurred at all stations about 3 weeks after treatment. This rise, coinciding with the decomposition of extensive masses of Chara, was probably caused by release of copper from its tissues. After these secondary peaks, a rapid decrease in dissolved copper levels oc- curred. Detrital particles from the decomposing Chara may have aided in removing copper from solution. FIGURE 4. Changes in dissolved and suspended copper concentrations, Pond B, three sites NOTE: Shaded area denotes treatment with 3 ppm copper sulfate. Suspended copper concentrations in Pond B increased slightly after treatment and then rose to levels between 0.3 and 0.5 mg/ liter after 21 days. Copper-bearing par- ticles from disintegrating Chara may have caused the elevated levels. In general, a rapid loss of dissolved copper from solu- tion has occurred in most experiments performed in alkaline water (5,] 3). In the present study, significant levels of dissolved copper remained after several weeks, possibly because of copper-containing organic com- plexes in the pond water. 228 SEDIMENTS After treatment in Pond A, copper accumulated in sedi- ment rapidly at first and then gradually (Fig. 5). Pre- cipitation of insoluble compounds probably accounted for the initial increase. Later accumulations may have been caused by gradual adsorption of dissolved copper and of copper released from plants and algae. Although sediments collected copper more quickly in Pond B than in Pond A, overall accumulation patterns of the two ponds were similar. Published research indicates that copper is rapidly adsorbed by pond sediments and that the amount of copper fixed is determined by the amount of organic matter and nature of clay minerals present (7). Because sediment in the current study contained a high level of organic matter (11 percent) copper was probably fixed once it reached the bottom. Evidence indicates that sediment acts as the ultimate repository of copper added to an aquatic system. Pre- cipitation of copper compounds followed by gradual adsorption of copper complexes from water probably occurred in the present study. 10000 - .o^. 1000 • • • • •. • • •' t z 5 100 _ • s •: • * < ' *' < 10 [- • o • • • i 1 ' B-17 9-6 9-26 10-16 11-6 11-26 DATE FIGURE 8. Copper accumulated by plant tissues, Pond B NOTE: Shaded area denotes treatment with 3 ppm copper sulfate. estimated amount of copper in sediment at the end of the experiment was greater than 90 percent of the total applied. _j i_ 7-20 8-12 9-1 10-5 10-25 11-14 DATE FIGURE 9. Copper accumulated by green sunfish. Ponds A and B NOTE: Shaded area denotes treatment with 3 ppm copper sulfate. COMPONENTS OF COPPER DISTRIBUTION The fraction of applied copper in each major compo- nent of Ponds A and B was estimated on each sampling date. Plant values were calculated by estimating the weight of plant masses and total copper accumulated in the masses. Water concentrations included copper in dissolved and suspended states. Weight of copper on the total sediment surface at each sampling time was estimated by multiplying the average copper value of an individual core sample by a factor calculated by dividing total sediment surface area by core surface area. Figure 10 graphs only water and sediment copper for Pond A because plant tissues always contained less than 1 percent of the total copper applied. A clear relation existed between water and sediment copper fractions in Pond A: a decrease of copper in water with a con- comitant increase in sediment copper occurred. The FIGURE 10. Total applied copper present in water and sediment. Pond A 230 Pesticides Monitoring Journal Plant tissue, mostly Chara, played a significant role in Pond B copper dynamics (Fig. II). Percentage of cop- per in plant tissues 14 days after treatment was esti- mated to be about 10 percent of the total applied. Copper was released shortly thereafter during decom- position. Sediment accumulation followed a pattern sim- ilar to that of Pond A. Copper values in the water were influenced by release of copper from dying Chara tissue. Conclusion As expected, copper moved rapidly from water to sedi- ment in the ponds observed in this study. However, phenomena such as binding of metals by organic com- pounds in the water and rapid uptake of metals by aquatic plants and algae cannot be overlooked in as- sessing dispersal of a metal in aquatic systems. LITERATURE CITED (1) Kopp, ]. F., and R. C. Kroner. 1967. A five-year sum- mary of trace metals in rivers and lakes of the United States. U.S. Dep. Interior; Compilation of Data, Oc- tober 1, 1962— September 30, 1967. 28 pp. (2) Bartley, T. R. 1969. Copper residue on irrigation canal. Rept. Bur. Reclamation, U.S. Dep. Interior. 16 pp. (3) Nelson, J. L., V. F. Brims, C. C. Coutant. and B. L. Carlile. 1969. Behavior and reactions of copper sulfate in an irrigation canal. Pestic. Monit. J. ^(3 ): 186-189. (4) Chancellor, R. J., A. V. Coombs, and H. S. Foster. 1958. Control of aquatic weeds by copper sulphate. Proc. 4th Brit. Weed Cont. Conf. Pp. 80-84. (5) Deuberl, K. H., and I. E. Demoranrille. 1970. Copper sulfate in flooded cranberry bogs. Pestic. Monit. J. 4(1):11-I3. (6) Toth, S. J., and D. N. Riemer. 1968. Precise chemical control of algae in ponds. J. Amer. Water Works Ass. 60(3):367-371. (7) Riemer, D. N., and S. J. Toth. 1970. Adsorption of copper by clay minerals, humic acid and bottom muds. J. Amer. Water Works Ass. 62(3 ): 195-197. (8) Riley, G. A. 1939. Limnological studies in Connecticut. Ecoi. Monogr. 9(1) : 54-94. (9) American Public Health Association. 1965. Standard Methods for the Examination of Water and Waste- water. 12th ed. New York. 873 pp. (10) U.S. Environmental Protection Agency. 1971. Methods for Chemical Analysis of Water and Wastes. Water Quality Office. Cincinnati, Ohio. 312 pp. (//) Stiff, M. J. 1971. The chemical states of copper in pol- luted fresh water and a scheme of analysis to differ- entiate them. Water Res. 5:585-599. (12) Ong, H. L., V. E. S»anson, and R. E. Bisque. 1970. Natural organic acids as agents of chemical weathering. U.S. Geol. Survey Prof. Paper 70O-C:C-I30-C-137. (13) Mulligan, H. F. 1969. Management of aquatic vascular plants and algae. In Eutrophication: Causes, Conse- quences, Correctives. National Academy of Sciences, Washington, D.C. 661 pp. FIGURE 1 1. Percent of total applied copper present in water, sediment, and plant tissue. Pond B Vol. 8, No. 4, March 1975 231 RESIDUES IN FOOD AND FEED Preliminary Survey of Ethylenethiourea Residues in the Canadian Food Supply, 1972 Zigmund Pecka,' Patricia Baiilu,- and Harvey Newsome 3 ABSTRACT A preliminary monitoring program was initiated in 1972 to determine ethylenethiourea (ETU) content of the Canadian food supply. Of 167 samples analyzed, 90 were domestic and 77 were imported. Samples were analyzed by electron- capture/gas-liquid chromatography. Thirty-three percent of the samples contained delectable ETU residues; most of these were 0.020 ppm or less. Highest levels, 0.047 and 0.083 ppm, were found in canned spinach and orange peel, respectively. Introduction Ethylenebisdithiocarbamates are widely used as fungi- cides and may. under conditions of aeration (!) or cooking (2). degrade to ethylenethiourea (ETU). The compound has been identified as a component of com- mercial ethylenebisdithiocarbamate formulations (3.4). Toxicological studies have shown ETU to be goitro- genic (5), carcinogenic (6). and teratogenic (7). In view of these findings, the present survey was conducted in 1972 to establish ETU levels in the Canadian food supply. Sampling Procedures A total of 167 samples were analyzed. Of these, 85 were obtained by the Inspection Services of the Quebec Re- gion, Health Protection Branch, and 82 were obtained by the Inspection Services of the Ontario Region. Both domestic and imported products were analyzed: 90 of the former and 77 of the latter. Of the 77 imports. 56 were from the U.S.A.. 9 from Mexico. 5 from Hol- land. 2 from Switzerland. 2 from Chile. 1 from Moroc- co. 1 from Israel, and 1 from Poland. ' Quebec Regional Laboratory, Health Protection Branch, Derartmenl of National Health and Welfare. 1001 St-Laurent, Longueliil, Quebec. Canada. = Ontario Ministry of the Environment, Toronto, Ontario, Canada. 3 Food Research Laboratories, Health Protection Branch. Department of National Health and Welfare, Ottawa, Canada. For small fruits and vegetables (e,g., grapes, Brussels sprouts, beans), minimum sample size was 3 lb. For medium-size fruits and vegetables (e.g., apples, oranges, potatoes), minimum samples were 3 lb or 10 units. For large commodities (e.g.. cabbage, lettuce), a minimum number of 3 was used for each sample. For canned goods, 5 cans of each type were combined to make 1 sample. A 1-lb minimum of cereal or wheat product was obtained for each sample. All samples were cut, blended, and stored in mason jars in a freezer before analysis. A nalytical Procedures The method of analysis was a modification of one described previously (5). Stored samples were thawed and a 5.0-g portion was homogenized with 50 ml abso- lute ethanol in a Sorvall Omni-Mixer. Solids were re- moved from the homogenate by filtration through Whatman No. 1 paper using a slight negative pressure and a filtrate diluted to 100 ml with distilled water. Twenty-ml aliquots of the diluted extract were placed in 50-ml round-bottomed flasks, and 0.1 ml benzyl chloride was added. The benzyl chloride had been puri- fied previously by passage through a column of alumina as described by Onley et al. (9) for n-bromobutane. After refluxing the contents of the flasks for 30 min- utes samples were cooled and transferred to 1 25-ml separatory funnels with 30 ml distilled water. Hydro- chloric acid (1 N\ 1.0 ml) was added and samples were extracted with two (1 X 10 ml and 1 X 5 ml) solutions of chloroform which were later discarded. Potassium hydroxide (I N\ 5.0 ml) was added to the aqueous phase and the s-benzyl ETU was immediately extracted with 10 ml chloroform. Extracts were dried by passage through a small bed of sodium sulfate and placed in 12-ml vials containing 10 |^il paraffin oil. Chloroform was removed by evaporation under a stream of nitro- 232 Pesticides Monitoring Journal gen. By varying nitrogen flow, the rate of evaporation was controlled enough that 1.5 hours or more were required to remove 10 ml. More rapid evaporation sometimes leads to losses of derivative. Samples were trifluoroacetylated by adding a solution of 10 percent trifluoroacetic anhydride in 0.50 ml ben- zene and permitting them to react for 15 minutes at room temperature. The solvent was evaporated almost to dryness under a gentle stream of nitrogen and the trifluoroacetylation was repeated with an additional 0.50 ml reagent for 1 5 minutes. After removal of the solvent, samples were dissolved in 1.0 ml benzene and a 5.0-^(1 aliquot was analyzed by gas-liquid chromatography (GLC). GLC was performed on a Varian Aerograph 1400 fitted with a tritium foil electron-capture detector and a 6-ft- by-Vs-in.-ID glass column. The column was packed with 4 percent SE-30, 6 percent QF-1 on Chromosorb W HP and operated at 200° C with a nitrogen flow of 100 ml/min. The injection port and detector were main- tained at 220° C. Under these conditions trifluoro- acetylated s-benzyl ETU had a retention time of 12 minutes. ETU, added at levels of 0.05 and 0.10 ppm to 17 com- modities, was recovered at a mean of 96.0 percent ± a standard deviation of 8.9 percent. The minimum detect- able limit, defined as twice background, was 0.01 ppm. Because of low levels of ETU encountered in the sam- ples, GLC/mass spectral confirmation (8) was not attempted. Results and Discussion Results of the analyses are presented in Table 1. Of the 167 composite samples analyzed for ETU residues, 112 samples (67 percent) did not contain detectable levels and 55 samples (33 percent) contained residues ranging from 0.01 to 0.15 ppm. Most of the samples (92 percent) contained 0.02 ppm or less ETU. Notable exceptions were some samples of canned spinach with an average of 0.047 ppm and orange peels with an average of 0.083 ppm. Of the 90 domestic samples, 29 composites (32 percent) contained detectable levels of ETU compared to 26 composites (34 percent) of the 77 imported foods containing detectable ETU levels. Acknowledgments Authors wish to thank K. A. McCuUy, Field Opera- tions Directorate, Health Protection Branch, for orga- nizing this project; G. Leveille, Health Protection Branch, Montreal, Canada, for advice in preparing this report; and C. Fortin and R. Gauthier, Health Pro- tection Branch, Montreal, Canada, for technical assis- tance. TABLE 1. Elhxienethiourea residues in foods sampled from Quebec and Ontario, Canada— 1972 Food Potatoes Tomatoes Tomatoes Apples Lettuce Carrots Carrots Green beans Green beans White kidney beans Cabbage Cabbage Sauerkraut Broccoli Brussels sprouts Celery Cherries Spinach Spinach Cucumber Dill pickles Grapes Oranges (peel) Oranges (pulp) Wheat Flour Pufled wheat Shredded wheat Wheat germ Wheat germ flakes Wheat hearts Bran flakes Cereals Type Fresh Fresh Canned Fresh Fresh Fresh Canned Fresh Canned Canned Fresh Canned Canned Fresh Fresh Fresh Canned Fresh Canned Fresh Jarred Fresh Fresh Fresh Fresh Bagged Boxed Boxed Boxed Boxed Boxed Boxed Boxed No. Samples Analyzed 13 10 5 10 10 9 1 4 5 1 8 1 3 10 8 7 10 7 5 7 3 10 5 5 5 3 1 I 1 1 1 1 1 NOTE: ND = none detected. Canned implies use of tin can; jarred implies use of glass jar. Vol. 8, No. 4. March 1975 No. Positive Samples 4 3 4 4 3 5 0 0 2 0 1 0 0 4 2 4 0 2 3 1 0 5 3 3 2 0 1 0 0 0 0 Mean of Positive Samples, ppm 0.025 0.020 0.018 0.028 0.013 0.010 ND ND 0.020 ND 0.010 ND ND 0.020 0.020 0.015 ND 0.025 0.047 0.050 ND 0.020 0.083 0.020 0.015 0.040 ND 0.010 ND ND 0.010 ND ND Range of Positive Samples, ppm 0.010.O.050 0.010-0.030 0.010-0.020 0.020-0.050 0.010-0.020 0.010-0.010 ND ND 0.020-0.020 ND ND ND ND 0.010-0.040 0.020-0.020 0.010-0.020 ND 0.020-0.030 0.040-0.050 ND ND 0.010-0.050 0.030-0.150 0.020-0.020 0.010-0.020 ND ND ND ND ND ND ND ND 233 LITERATURE CITED (/) Ludwig, R. A., G. D. Thorn, and D. M. Miller. 1954. Studies on the mechanism of fungicidal action of di- sodium ethylenebisdithiocarbamate (Nabam). Can. J. Bot. 32:48-54. (2) Newsome, W. H., and G. W. Laver. 1973. Effect of boiling on the formation of ethylenethiourea in zineb- treated foods. Bull. Environ. Contam. Toxicol. 10(3): 151-154. (i) Johnson, E. I., and J. F. C. Tyler. 1962. Occurrence of ethylenethiourea in thiocarbamate fungicides and its detection in fruit juice. Chem. Ind. (London) 305- 306. {4) Bonloyan, W. R., J. B. Looker, T. E. Kaiser, P. Giang, and B. M. Olive. 1972. Survey of ethylene- thiourea in commercial ethylenebisdithiocarbamate formulations. J. Ass. Offic. Agr. Chem. 55:923-925. (5) Graham, S. L., and W. H. Hansen. 1972. Effects of short term administration of ethylenethiourea upon thyroid function of the rat. Bull. Environ. Contam. Toxicol. 7:19-25. (6) Meland, B. M., 1. H. Weisburger, E. K. Weisburger, J. M. Rice, and R. Cypher. 1972. Thyroid cancer in rats from ethylene thiourea intake. J. Nat. Cancer Inst. 99:583-584. (7) Khera, K. S. 1973. N,N'-ethylenethiourea: teratogenic- ity study in rats and rabbits. Teratology 7:243-252. (8) Newsome, W . H. 1972. Determination of ethylene- thiourea residues in apples. J. Agr. Food Chem. 20: 967-969. (9) Onley, ]. H., and G. Yip. 1971. Determination of ethylene thiourea residues in food, using thin layer and gas chromatography. I. Ass. Offic. Anal. Chem. 54(1): 165. 234 Pesticides Monitoring Journal RESIDUES IN FISH, WILDLIFE, AND ESTUARIES Pesticide and Mercury Residues in Commercially Grown Catfish ^ A. B. Crockett,2 G. B. Wiersma.s h. Tai,3 and W. Mitchell 3 ABSTRACT In 1970, 54 commercial catfish farms in Arkansas and Mis- sissippi were sampled for pesticide and mercury residues. Pesticide residues above FDA action levels were detected in 15 percent of the fish samples. Data on residues in sedi- ment, fish feed, and source water suggest that fish were not being contaminated from these sources. Average fish residue per county was, however, strongly correlated with the per- cent of total acres planted in cotton and soybeans. Results strongly suggest that cotton production was the primary source of contamination. Actual routes of movement have not been clearly defined but aerial transport seems most probable. Introduction In February 1970 the Plant Protection Division, Agri- cultural Research Service, U.S. Department of Agricul- ture, initiated a study to determine the magnitude and source of pesticide residues in commercially grown cat- fish. The study was undertaken to investigate reports of illegal residues of DDT, endrin. and dieldrin in com- mercially raised catfish of the Mississippi delta, which is the major catfish-growing area of the United States. Analyses for mercury were included for scientific in- terest. Although little information is available on pesticide resi- dues in cultured catfish, it is generally known that fish can concentrate chlorinated hydrocarbon pesticides. 1 Technical Services Division, Office of Pesticide Programs, U.S. En- vironmental Protection Agency. Washington, D.C. 20460. 2 National Environmental Research Center, U.S. Environmental Pro- tection Agency, Box 15027, Las Vegas, Nev. 89114. Reprints avail- able from this address. ' Technical Services Division, Office of Pesticide Prograins, U.S. En- vironmental Protection Agency, Mississippi Test Facility, Bay St. Louis, Miss. Bevenue et al. U) reported that chlorinated pesticides can be concentrated to 33,000 and 36,000 times the level found in the water by carnivorous and detrital fish, respectively. Morris and Johnson (2) studied the dieldrin level of fish in Iowa streams and found that catfish contained higher pesticide levels than other rough fish and much higher concentrations than game fish. A study of wild catfish in Nebraska revealed the frequent presence of DDTR (DDT + DDE -f TDE) and diel- drin (i). Channel catfish have been included regularly as a pre- ferred species for sampling in the National Pesticides Monitoring Program for fish because they are near the top of the food chain (4). The two monitoring sites located in the commercial fish-raising area of Arkansas consistently reported endrin residues during the spring and fall of 1967 and 1968 (5). DDT residue levels in fish from these two areas are among the highest detected in the Mississippi River System (6). Sampling Procedures Sampling was conducted in Arkansas and Mississippi because those States encompass the most intensive cat- fish farming area in the United States. Fifty catfish farms were selected on a probability basis: i.e.. each farm had an equal chance of being selected. Approxi- mately 1 farm in 12 was examined. At each farm, fish, source water, pond water, and sediment were sampled from one pond; a feed sample was also collected. Sam- ple collection commenced in March 1970 and was completed by the end of April. Catfish samples were composed of two or three fish, each about 2 years old and weighing 0.5-0.7 kg. As Vol. 8, No. 4, March 1975 235 specified in the U.S. Department of Health, Education, and Welfare Food and Drug Administration regulatory methodology, only edible fish flesh was examined for pesticide residues. One gallon of source water and a composite gallon sample of pond water collected from 5 to 10 different pond locations were taken from each farm. One composite sediment sample was collected from each pond with a weighted dredge. At least five drag subsamples were taken within 10 feet of shore; another five drags were taken farther from shore. A sample of catfish feed was collected: when possible, from different feedbags or lots. Analytical Methods SEDIMENT After decanting and discarding excess water, the remain- ing portion was mixed for 5 minutes in a paint shaker and a 300-g sediment sample was taken. The sample was extracted with 600 ml 3:1 hexane:isopropanol by con- centric rotation for 4 hours. Alcohol was removed by three water washes and the hexane extract was dried through anhydrous sodium sulfate. It was then ready for gas-liquid chromatographic (GLC) analysis. A sep- arate portion of sediment was dried at 120°C for 24 hours to obtain a moisture content value. Analytical data on sediment samples were calculated on a dry- weight basis. WATER The gallon water sample was shaken before removal of a 500-g subsample. The sample contained some sus- pended matter and was not filtered before extraction. The 500-ml sample was extracted three times with 60 ml methylene chloride by shaking in a 1,000-ml sepa- ratory funnel. The three extracts of about 180 ml were combined and concentrated to about 5 ml, 100 ml hex- ane was added, and the sample was concentrated again to 5 ml. All concentrations were performed under a three -ball Snyder column except the final adjustment to 2.5 ml which was performed in a centrifuge tube with a gentle stream of air. At this point the sample extract was ready for analysis. TISSUE. ALGAE, AND FEED SAMPLES Feed samples were received in 1 -gallon and Vi -gallon cans and thoroughly mixed to give a representative sample. Catfish were beheaded, skinned, and eviscerated and the meat and bone were thoroughly macerated in a Hobart food grinder. A 20-g sample was mixed with 100 ml isopropanol for 2-3 minutes in a Waring blender. Three hundred ml hexane was added and the sample was rotated concen- trically for 2 hours. An aliquot representing 15 g was taken, the isopropanol was removed by two water washes, and the hexane was extract-dried through so- dium sulfate. Algae and feed were processed in the same manner as tissue except that 100-g samples of feed were used. Partitioning of Samples — An aliquot of 15 g was parti- tioned as follows: the 50-ml hexane sample extract was shaken with 100 ml acetonitrile in a 500-ml separatory funnel, the bottom acetonitrile layer was saved, and the hexane layer was discarded. This step was carried out three times and the acetonitrile layers were combined. The combined acetonitrile extracts were backwashed with 25 ml acetonitrile-saturated hexane and the hexane layer was discarded. The acetonitrile sample extract was concentrated to approximately 10 ml under a three-ball Snyder column and 100 ml hexane was added. The latter two operations were repeated twice and the sam- ple was essentially in hexane. The hexane extract was adjusted to appropriate volume and held at low tem- perature for subsquent florisil column cleanup and frac- tionation. Cleanup and Fractionation — A chromatographic column consisting of a 125-ml flask reservoir attached to an ll-by-500-ml glass tube with a teflon stopcock and a removable glass tip was prepared by placing a small pad of hexane-washed glass wool in the bottom of the column and adding anhydrous granular sodium sulfate to the 1-inch level. Eighteen g 60-120 mesh florisil was poured into the column and evenly packed by tapping the column with a light mallet. The column was prewashed with 100 ml nanograde hexane and the sample extract representing 5 g of the original sample was quantitatively added to the column when the level of the hexane prewash had reached the top of the upper layer of Na^SO^. When the sample extract level reached the top of the column, 100 ml 5 percent methylene chloride in hexane was added and a 250-ml flask marked Fraction 1 was placed under the column. When the liquid level again drained to the top of the column, 100 ml nanograde methylene chloride was added to the reservoir and the original flask was replaced by a second, marked Fraction 2. One ml 0.01 percent Nujol in hexane was added to each elution. Each elution was concentrated to approximately 5 ml under a Snyder column. One hundred ml hexane was added to each elution and the fractions were again concentrated to approximately 5 ml. Each elution was quantitatively transferred to a 15-ml centrifuge tube and placed in a 40°C water bath. A gentle stream of air was directed into the tubes and the sample volume was reduced to 2.5 ml. Fractions were injected separately onto a gas-liquid chromatograph. Gas-Liquid Chromatographic Analysis — At least two columns were used for each sample; a third column was also used when necessary. Operating parameters varied for the three columns: 236 Pesticides Monitoring Journal Column 1. 4.8 percent OV-17/6.2 percenl QF-1 on Gas-Chrom Q Temperatures: Injector 250° C Oven 200° C Detector 210° C Column 2. 3 percent DC-200 on Gas-Chrom Q Temperatures: Injector 245° C Oven 175° C Detector 205° C Column 3. 9 percent QF-1 on Gas-Chrom Q Temperatures: Injector 230° to 245^ C Oven 70° C Detector 200° to 205° C Carrier gas was 5 percent methane in argon or pre- purified nitrogen at 80-100 ml/min. Recoveries were performed for each type of sample. Recoveries were consistent with values from the same type of samples in previous work completed in this laboratory. Chlorinated pesticides used for recovery studies were heptachlor, heptachlor epoxide, gamma chlordane, o,p'- DDE, p.p'-DDE, dieldrin, endrin. o,p'-DDT. p.p'-DDT. p./^'-TDE. and aldrin; organophosphorus compounds used were phorate, diazinon, methyl parathion. ethyl parathion, DEF, trithion, and ethion. Recovery was 90 to 100 percent on water and sediment, and 80 percent on tissue, algae, and feed. Toxaphene was quantitated by comparing the four major peaks with corresponding peaks on a calibrated stan- dard chromatogram whenever possible. In some cases only two peaks could be compared with corresponding peaks from the standard. Chlordane was quantitated by use of the gamma chlordane peak or. if necessary, by comparison with a technical chlordane standard. Quan- titations were confirmed by dual-column or triple-col- umn cross-checking and use of p-values (7). Limits of Detection — Minimum detection limts (MDL) in fish, algae, feed, and sediment were 0.01 ppm for all pesticides except mixtures such as chlordane. tox- aphene. polychlorinated biphenyls (PCB's). and a few compounds such as EPN or Guthion, which elute later on GLC columns. In these cases, the MDL was 0.03 to 0.10 ppm, depending on the compound and the noise level. Corresponding limits for water were 0.01 or 0.03 to 0.10 ppb. Mercury Analysis — A sample of mercury weighing approximately 2 g was placed in a 250-ml Erlenmeyer flask and 25 ml concentrated sulfuric acid was added. The sample was heated until the organic matter had dissolved. Organic matter was further oxidized by add- ing 1 .0 ml 30 percent hydrogen peroxide until the sam- ple was colorless. Fifty ml distilled water was added to the sample. After cooling, 5 percent potassium per- manganate was added, one drop at a time, until a per- manent pink color was attained. Twenty ml sodium chloride — hydroxylamine sulfate was added and the flask was positioned in the aeration sys- tem. Ten ml stannous sulfate was added and the mer- cury was swept into the ceil as elemental mercury. The absorbance reading of the mercury was compared with a standard curve plotted from known amounts of mer- cury which had been analyzed by the same methods applied to the samples. The MDL was 0.01 ppm. This method of analysis was patterned after that of Hatch andOtt (S). Results AH fish examined showed pesticide residues. Results of the fish analyses (Table 1 ) indicate that DDTR and mercury were found in all samples; dieldrin, endrin, and toxaphene were present in 89, 76, and 96 percent of the samples, respectively. Chlordane was found once and aldrin existed at low levels in three samples. No other chlorinated hydrocarbon pesticides were detected. Although most fish samples contained residue levels below the FDA action level or tolerance, a number of samples exceeded the limit. FDA action levels were ex- ceeded by 2 percent of the DDTR samples, 6 percent of the aldrin/dieldrin samples. 4 percent of the endrin samples, and 7 percent of the toxaphene samples. In total, 1 5 percent of the fish samples exceeded present limits for one or more residues. Toxaphene appears to be the most serious contaminant because it exceeded the action level most frequently and its average con- centration was closest to its limit. Average concentra- tion of the four samples exceeding the 5 ppm limit was 13.0 ppm. Possible sources of pesticide residues in fish would ap- pear limited to source water, sediment, feed, and crops. While it is conceivable that stocked fingerlings were a source of pesticides, no evidence in the study indicates this to have been the case. All the farmers claimed that TABLE I. Pesticide and mercury residues in commercial catfish No. Samples Percent with Residues Average Concentration, PPM FDA Action Limit, ppm Percent Samples Exceeding Limit Range Detected Residues, ppm DDTR Aldrin/Dieldrin Endrin Toxaphene Mercury 54 54 54 54 50 100 89 76 96 100 1.07 0.07 0.06 2.1 0.07 5 0.3 0.3 5 0.5 n. 09-8. 71 0,01-0.87 0.01-0.41 0.2-20.7 0.02-0.35 NOTE: Samples represent edible portions of catfish. Vol. 8. No. 4. March 1975 !37 they had not applied any of the contaminating pesti- cides to the fish ponds. WATER Thirty-five samples of pond and source water were examined for chlorinated hydrocarbons. In no instance was any pesticide detected at the 0.01 ppb level. No fish from wells were analyzed for chlorinated hydrocarbons because these compounds are insoluble in water and are not leached to any appreciable extent. To determine whether lower-level contamination existed, residues in catfish were statistically compared according to the water source in which the fish had been caught. A comparison of log-transformed means of DDT, diel- drin, endrin. and toxaphcne residues was made between catfish raised in well water and those raised in surface water. Well-water catfish had significantly higher pesti- cides (95 percent level) in all cases. These data do not, however, indicate that the pesticide came from wells but, rather, that the pesticide source was associated with wells. Further explanation appears under the crop section of this pajDcr. During a resampling of some of the fish ponds, several algae samples were collected and analyzed. All five samples showed DDT and toxaphene residues. DDTR and toxaphene levels ranged from 0.10 to 0.97 and from 0.20 to 1.41 ppm, respectively. The absence of chlorinated hydrocarbons from pond water can prob- ably be explained by their low solubility in water. Algae residues, however, indicate that pesticides must have been in the water at some time. Surface runoflf could be a source of f)esticide residues but the absence of residues from sediment, which is also transported in runoff, does not support this theory. In addition, commercial fish ponds are generally con- structed to avoid surface runoff if clean water is avail- able. SEDIMENT Fifty-three sediment samples were examined for chlo- rinated hydrocarbons. The only chemical found was DDTR. which was present in 18 samples. A correlation analysis of all samples was run to estimate the level of association between DDTR in fish and DDTR in sedi- ment. Results showed a correlation coefficient (r) of 0.287, which indicates a significant deviation from zero at the 95 percent level. If DDTR were derived from the original soil prior to pond formation, the DDT level probably would decrease as the pond aged. No signifi- cant difference from zero could be established. Another consideration is that DDTR and toxaphene in fish are closely correlated (99 percent level, r = 0.60, n = 54), which suggests they are derived from the same source. The absence of toxaphene from sediment samples sug- gests that sediment is not the source of fish residues for this insecticide. The absence of pesticide residues from sediment sam- ples could also indicate poor sampling techniques. In the resampling of ponds. DDTR was the only pesticide found. Toxaphene, however, was found in algae which eventually becomes detritus and enters the sediment. Absence of residues could mean that fine surface sedi- ment was not adequately sampled. F EED All fish feed from the 43 farms sampled had detectable residues (Table 2). In addition to pesticides shown in Table 2, small amounts of heptachlor, heptachlor epox- ide, and Aroclor 1254 were detected in a few samples. A correlation analysis did not indicate a significant asssociation between DDTR. dieldrin. endrin, and tox- aphene levels in feed and fish (Table 2). In addition, DDTR and toxaphene residues, which correlate closely in fish, are not correlated in feed (r = 0.200). Pesticide residues in feed certainly contribute to the residues found in fish, but the amount appears negligible com- pared with other sources. CROPS To determine whether pesticides used on crops had con- tributed to contamination of catfish, authors compared the percent of total county acres to which insecticides had been applied (9) with the average fish residue for that county. Resulting correlation coefficients indicate a significant deviation from zero (99 percent level) in TABLE 2. Principal pesticide residues in fish feed No. Samples Percent with Residues Average Concentration, PPM Range Detected Residues, ppm Correlation COEFFICIEN r. Fish: Feed DDTR 43 91 0.120 0.02-0.84 0.005 Dieldrin 43 74 0.007 0.01 0.166 Endrin 43 14 0.002 0.01-0.02 -0.149 Chlordane 43 21 0.016 0.01-0.28 ND Toxaphene 43 42 0.06 0.1-0.3 0.099 Malathion 43 74 0.051 0.01-0.32 ND NOTE: n = 43, r = 0.301 at 95% confidence level; ND r^ data insufficient to calculate. r = 0.391 at 99% confidence level. 238 Pesticides Monitoring Journal TABLE 3. Correlations between land use practices and insecticide residues in catfish Acres Receiving Insecticide, % Acres IN Harvested Crops, % Acres IN Cotton, % Acres IN Soybeans, % Mississippi' DDTR Corr. Coet. 0.794 0.701 0.830 0.636 95% Conf. Inlerval 0.551-0.913 0.386-0.870 0.621-0,929 0.281-0.838 DIELDRIN Corr. Coef. 0.662 0.587 0.706 0.590 95% Conf. Interval 0.323-0.851 0.208-0.813 0.395-0.872 0.213-0.814 ENDRIN Corr. Coef. 0.732 0.690 0.818 0.587 95% Conf. Interval 0.439-0.884 0.369-0.864 0.596-0.923 0.208-0.813 TOXAPHENE Corr. Coef. 0.687 0.675 0.767 0.563 95% Conf. Interval 0.363-0.863 0.343-0.857 0.502-0.901 0.174-0.800 Arkansas= DDTR: Corr. Coef. 0.295 ^0.285 -0.211 -0.238 DIELDRIN: Corr. Coef. -0.007 0.176 0.143 0.170 ENDRIN: Corr. Coef. 0.547 -0.195 -0.046 -0.171 TOXAPHENE: Corr. Coef. 0.197 -0.019 -0.021 0.024 Mississippi but not in Arkansas (Table 3). Similar re- sults were obtained when the percent of total county acres in harvested crops (9) was compared with aver- age fish residues. The major crops and primary recipients of insecticides in Arkansas and Mississippi are soybeans and cotton (9). In the period preceding this study, DDT, endrin, dieldrin, and toxaphene were registered for use on cot- ton. Of these insecticides, only DDT and toxaphene had been used for more than seed treatment of soybeans. Frequently a combination of DDT and toxaphene had been used several times a year on cotton but usually soybeans had been treated just once every few years to control insect outbreaks (W). Correlations comparing the percent of total county acres in cotton or soybeans in Mississippi and average insecticide residues in fish showed significant deviation from zero at the 99 percent level for both crops (Table 3). Correlation coefficients for cotton and soybeans were not significantly different, but cotton always re- sulted in a more significant deviation from zero for each insecticide. Correlations in Arkansas were again not significant. The absence of correlation in Arkansas may be due to two factors. Most Arkansas sites sampled were located in the highly agricultural eastern counties, but in Mis- sissippi, sites were scattered throughout the State. Only in Mississippi, therefore, could agricultural areas be compared with nonagricultural areas. A second reason for the insignificant correlations in Arkansas may be that the northeastern counties have fewer insect prob- lems and generally apply a smaller amount of pesticides than do the southeastern counties. Because Mississippi data were considered more informa- tive, they were chosen for further examination. Missis- sippi data were segregated into high-cotton areas (>10 percent of the total county acres in cotton) and low- cotton areas (<10 percent of the total county acres in cotton). Mean pesticide levels in catfish were calculated after logarithmic transformation; both cotton areas were compared using a 95 percent confidence interval about the mean. Since residue data were not normally distributed, log transformation was used to help normal- ize them. Results indicate that fish in high-cotton areas have sig- nificantly higher concentrations of DDTR, endrin, and toxaphene than they have in low-cotton areas (Table 4). This relationship could account for the high cor- relation between DDT and toxaphene in fish, for DDT and toxaphene had often been applied together on cotton. TABLE 4. Mean pesticide residues and 95 percent con- fidence intervals for fish from high-cotton versus low-cotton areas, Mississippi DDTR Dieldrin Endrin Toxaphene Residues, ppm Geometric mean, high- cotton area 1.48 0.042 0.063 2.73 Geometric MEAN, LOW- COTTON AREA 0.34 0.014 0.010 0.42 95% CONF. Interval, high-cotton AREA 2.40 -0.91 0.090-0.017 0.122-0.030 4.42 -1.68 95% Conf. Interval, low-cotton AREA 0.53 -0.22 0.024-0.007 0.019-0.005 0.85 -0.21 High pesticide levels found in fish grown in well water can also be explained by the association between wells and high-cotton areas: 16 of the 19 farms in Mississippi which use wells as the sole water source are located in high-cotton counties. Residues in sediment were also related to high-cotton counties. Of 12 sediment samples with pesticide residues, 1 1 were located in such counties. Vol. 8, No. 4, March 1975 239 Pesticide residues in catfish are closely correlated with crops and pesticide use. Considering that larger amounts of pesticides are applied to cotton than to any other crop in this area of the South, this study strongly sug- gests that cotton is the primary source of contamination in fish ponds. Data indicate that neither source water nor surface runoff appear to be the mechanism of pesti- cide transport. These data and the general absence of pesticide residues in sediments indicate that aerial trans- port of pesticides may be the major route of catfish contamination. LITERATURE CITED (/) Bevemie, A., J. W. Nylin, Y. Kawano, and T. W. Kelley. 1972. Organochlorine pesticide residues in water, sediment, algae and fish, Hawaii — 1970-71. Pestic. Monit. J. 6(l):56-64. (2) Morris, R. L., and L. G. Johnson. 197 1. Dieldrin levels in fish from Iowa streams. Pestic. Monit. J. 5(1): 12-16. (i) Stucky, N. P. 1970. Pesticide residues in channel cat- fish from Nebraska. Pestic. Monit. J. 4(2): 62-66. (4) Johnson, R. E., T. C. Carver, and E. H. Dustman. 1967. Residues in fish, wildlife, and estuaries. Pestic. Monit. J. 1(1):7-13. (5) Henderson, C, W. L. Johnson, and A. Inglis. 1969. Organochlorine insecticide residues in fish. Pestic. Monit. J. 3(3):145-I7I. (6) Henderson, C, A. Inglis, and W . L. Johnson. 1971. Organochlorine insecticide residues in fish — fall 1969. Pestic. Monit. J. 5(1): 1-11. (7) Bowman, M. C, and M. Beroza. 1965. Extraction p- values of pesticides and related compounds in six bi- nary solvent systems. J. Ass. Offic. Anal. Chem. 48(5):943-952. (8) Hatch, W. R., and W. L. Ott. 1968. Determination of submicrogram quantities of mercury by atomic ab- sorption spectrophotometry. Anal. Chem. 40(14): 2085-2087. (9) Bureau of the Census. United States Census of Agri- culture. 1969. Vol. 1, Section 2, Tables numbered 8 for Arkansas and Mississippi. {10) Ford, Rutledge F. 1973. Area Agronomist, U.S. De- partment of Agriculture, Extension Service, Ark. Per- sonal communication. 240 Pesticides Monitoring Journal Residues of Methoxychlor and Other Chlorinated Hydrocarbons in Water, Sand, and Selected Fauna Following Injections of Methoxychlor Black Fly Larvicide into the Saskatchewan River, 1972 ^ F. J. H. Fredeen, J. G. Saha, and M. H. Balba ABSTRACT In May 1972, 0.309 ppm methoxychlor black fly larvicide was applied in a single lest on the North Saskatchewan River. Eight to nine days later residues of 0.05-0.10 ppm methoxychlor occurred in sand 21-22 km downstream from the point of injection. Methoxychlor was not detected in water, insect larvae, shellfish, or muscle tissues of three fish species on the same sampling date. Perhaps because of rela- tively high oil content in goldeye fish, methoxychlor resi- dues in muscle tissues were 1.0-1.5 ppm in 8 percent of those sampled, 0.21-0.99 in 21 percent, and 0.02-0.20 in 37 percent. In 34 percent of the goldeye fish no residues were detected. Goldeye and other fish collected before or 17 weeks after this injection did not contain detectable levels of methoxychlor. River water in two samples of the injected slug of water collected 6.5 km downstream from the point of injection contained 0.14 and 0.16 ppm methoxychlor. The suspended solids filtered from these samples contained 40 and 47 percent of this methoxychlor (437 and 892 ppm, respectively). Thus methoxychlor may act selectively against filter-feeding species, especially black fly larvae. Introduction DDT black fly larvicide was injected once or twice into one or both branches of the Saskatchewan River during most years from 1948 through 1967. By 1968 residues in muscle tissues of fish from the river included up to 0.05 ppm DDT, 0.05 ppm DDD, and 0.06 ppm DDE (1). Methoxychlor black fly larvicide was injected experi- mentally on 1 1 occasions into the north and south branches of the river from 1968 through 1972 (2). 1 Contribution No. 530, Research Station, Canada Department of Agri- culture, Saskatoon, Saskatchewan, Canada. At the conclusion of these tests, and specifically in con- junction with a single test on the North Saskatchewan River in May 1972, samples of water, riverbed sands, clams, insect larvae, and fish were analyzed for meth- oxychlor residues. It is the single test in May 1972 from which most of the specific data in this paper have been compiled. Tests with single 15-minute injections of methoxychlor as a black fly larvicide commenced in the South Sas- katchewan River in 1968 and in the North Saskatche- wan River in 1969. The total number of 15-minute injections of methoxychlor at each of four sites is shown in Figure 1. A commercial 24 percent emulsi- fiable concentrate was used in seven of the eight experi- mental injections into the South Saskatchewan River and in all six 15-minute injections into the North Saskatchewan River; a commercial 50 percent wettable powder was used in one 1968 application in the South Saskatchewan River. A total of 285 kg technical meth- oxychlor was used in the eight injections into the South Saskatchewan; 450 kg was applied in the six injections into the North Saskatchewan. These were the only potential sources of methoxychlor residues in either branch of the Saskatchewan River up to and including May 23, 1972. That day marked the beginning of the study reported here, a study designed specifically to determine whether methoxychlor residues existed before or after a single 15-minute injection of 0.309 ppm methoxychlor into the North Saskatchewan River at Cecil Ferry (Fig. 1) on May 23. The injection period at Cecil Ferry lasted from 2:37 to 2:52 p.m. The injected mass of water passed the Cecil Rapids site about 4:40 to 5:15 p.m., as indicated by the solvent odor and by drogues that Vol. 8, No. 4, March 1975 241 had been released at the Cecil Ferry site at the leading and trailing edges of the treated mass of water. The treated water is assumed to have passed the Lacolle Falls site during the night of May 23-24. Figure 1 shows all three sites where samples were collected for analyses of residues. Also on May 23. 0.186 ppm methoxychlor was injected for 1 5 minutes into the South Saskatchewan River from Birch Hills Ferry. To- tal injected amounts of technical methoxychlor were 65.0 and 32.7 kg, respectively. FIGURE 1. North and Si I'lh Saskatchewan Rivers, cen- tral Saskatchewan, showing injection sites and numbers of methoxychlor hlacl: fly larricide injections, 1968-72 Prior to May 23, 1072, methoxychlor had last been injected into the river May 21 and June 4. 1971, when two 15-minute injections of 0.143 and 0.301 ppm were made from the Birch Hills Ferry and two 15-minute injections of 0.299 and 0.301 ppm were made from Cecil Ferry. Sampling Samples for residue analyses were collected only from the North Saskatchewan River, either from the Cecil Ferry crossing, site of the methoxychlor injection; from Cecil Rapids, 6.5 km downstream from Cecil Ferry; or from Lacolle Falls, 21-22 km downstream from Cecil Ferry (Fig. 1). Average width of the river was about 250 m. Its volume discharges were 236 m^/second on May 23, about 340 m'^/second on June 1, and about 140 m^/ second on September 19. The riverbed con- sisted of gravel and rocks interspersed with occasional beds of sand. There were numerous rapids including Lacolle Falls in this section of the river, all navigable by canop Water samples were collected only at midriver sites, either the Cecil Ferry crossing or Cecil Rapids (Table 1 ) ; samples were taken before, during, and after pas- sage of the injected water. These samples were col- lected by hand in Teflon-coated jars which were opened and filled beneath the water surface. Sand was collected beneath about 60 cm flowing water several meters from both river margins near Lacolle Falls. A hand shovel with 5-cm sides was guided along the riverbed to collect only the uppermost layer of sand to a depth of 0.5 cm. Mussels ranging from about 0.2 to 0.6 cm in diameter were washed out of sand collected from under 30-60 cm water along the north side of the North Saskatchewan River at TABLE 1. Methoxychlor residues in samples of water and sand collected from North Saskatchewan River, Mav-June — 1972 Date Collected Material Collected Methoxychlor Content, ppm River Water May 20: 2 samples Cecil Ferry Whole water ND Suspended solids ND Filtrate ND May 23: prior to Cecil Rapids Whole water ND arrival of treated Suspended solids ND water Filtrate ND May 23: 15 min after Cecil Rapids Whole water 0.160 arrival of leading edge Suspended solids 892.0 of treated water FUtrate 0.085 May 23: 30 min after Cecil Rapids Whole water 0.140 arrival of leading edge Suspended solids 437.0 of treated water Filtrate 0.084 May 30 Cecil Rapids Whole water ND Suspended solids ND Filtrate ND June 1 Cecil Rapids Whole water ND Suspended solids ND Filtrate ND Sand May 16 Lacolle Falls, N. side NA ND May 16 Lacolle Falls, S. side NA ND June 1 Lacolle Falls, N. side NA 0.10 June 1 LacoUe Falls, S. side NA 0.05 NOTE: ND = not detected (<0.01 ppm methoxychlor). NA = not applicable. 242 Pesticides Monitoring Journal Lacolle Falls. Odonata larvae about 1.5 cm long were also extracted from the sand along with the mussels. Trichoptera larvae about 1.0 cm long were collected from submerged rocks at midriver sites near Lacolle Falls. About 100 small mussels, 100 Odonata larvae, and 30 Trichoptera lirvae were collected on each sam- pling date. Black fly 'arvae were not available for this study. A nylon net wi h mesh openings of less than 0.1 mm was used to co'lect insects drifting in water at Cecil Rapids before and during passage of methoxychlor- injected water. A seine was used to collect fish at Lacolle Falls on May i6 but overnight gill net sets were used on May 17 and 31, June 1, and September 19. All samples of Gand, mussels, in.^ect larvae, and fish were wrapped immediately in aluminum foil and frozen in solid COo. They were stored at — 18° C until analysis. Analytical Procedures All solvents were pesticide analysis grade and were redistilled in glass. Water samples were analyzed within 24 hours of collection. Two-liter aliquots of thoroughly mixed water samples were filtered under suction to remove suspended solids. The filtrate was extracted with 200 ml hexane three times. The combined hexane extract was dried over anhydrous sodium sulfate, con- centrated to a suitable volume, and analyzed by electron-capture/gas-liquid chromatography (EC/GLC). Suspended solids were collected on filter paper, air dried, and weighed. Samples weighing about 0.2 g each were extracted by shaking for 1 hour with 25 ml 4:1 hexane: acetone mixture and filtered. Residues were reextracted twice with the same amounts of solvent mixture. The combined extract was dried over anhy- drous sodium sulfate and concentrated to about 10 ml. The concentrated extract was chromatographed on a 20-g florisil column containing 3 percent water and eluted with 200 ml 6 percent diethyl ether in petroleum ether, followed by 200 ml 16 percent diethyl ether in petroleum ether. Combined eluents were concentrated to a suitable volume and analyzed by GLC. All fish specimens were filleted and only edible muscle tissues without any skin were analyzed for pesticide residues. Fish specimens of the same species and similar sizes were first analyzed as composite samples. For this purpose 4- or 5-g muscles from 4 or 5 fish were pooled into 20-g samples: pesticide residue. levels were determined by the method described previously by Fredeen et al. (/). If any pooled samples showed residues of methoxychlor, all fish specimens represent- ing the composite were then analyzed separately by the same method. Mussels, drift net collections, and insect larvae samples were also analyzed by Fredeen's method although they were extracted with 5 ml acetonitrile for each gram of sample. Riverbottom material which consisted mostly of sand was partly air-dried and thoroughly mixed. Twenty-g aliquot samples were extracted with 1 :1 hexane: acetone by the method described earlier by Saha (3). Extracts were cleaned and analyzed as described by Fredeen et al. (7). GAS CHROMATOGRAPHY An Aerograph Hi-Fy gas chromatograph, Model 600D, was used for quantitative analysis. Operating conditions were: Column: Detectors: Temperatures: Carrier gas: Flow rate: Electrometer: Aluminum, 5 ft by '/e in. ID, packed with 4 per- cent SE-30 and 6 percent QF-1 on 80-100 mesh chromosorb W. Electron-capture, with tritium ionizing source. Column 185° C. Injector 200° C. Detector 200° C. Oxygen-free nitrogen. 40 ml/min Range 1 Sensilivitv 4 Under these conditions retention time for aldrin was 3.0 minutes. Retention times for other organochlorine insec- ticides relative to aldrin were: heptachlor, 0.94; hepta- chlor epoxide, 1.81; p,p'-DDE, 2.47; dieldrin, 2.81; p.p'-DDD, 3.25; p,p'-DDT, 3.78; and p.p'-methoxychlor, 7.25. All samples were analyzed for the above-mentioned organochlorine insecticides. Percent recoveries obtained when these chemicals were added at 0.10 and 0.25 ppm to 20-g samples of fish, extracted, and analyzed as above were: heptachlor, 78-92 percent; aldrin, 68-82 percent; heptachlor epoxide, 87-101 percent; p.p'-DDE, 68-85 percent; dieldrin, 84-102 percent; p.p'-DDD, 71-87 percent; p.p'-DDT, 86-98 percent; and p,p'- methoxychlor. 86-108 percent. The lower limit of de- tection was 0.01 ppm. Data reported below are not corrected for percent recovery. Any sample showing more than 0.1 ppm of any of the above organochlorine insecticides was also analyzed by thin-layer chroma- tography (TLC) according to the method of Kovacs (4). GAS CHROMATOGRAPHY/MASS SPECTROMETRY Presence of methoxychlor in excess of 0.2 ppm in gold- eyes was confirmed by gas-liquid chromatography/ mass spectrometry (GLC/MS). A Finnigan Model 3000 quadrupole mass spectrometer, connected to a Varian Aerograph Model 1400 gas chromatograph by means of an all-glass single-stage jet separator, was used. A S-ft-by-Vs-in.-ID glass column packed with 3 percent OV-1 on 60- to 80-mesh Gas Chrom Q was used for GLC separation. Column temperature was 210° C and helium flow rate was 15 ml/min. Mass spectra were recorded at 70 eV electron energy and sweep time of 10 seconds. Mass spectrum could be obtained when Vol,. 8, No. 4. March 1975 243 2.5 ^ig reference methoxychlor was injected into the gas chromatograph. About 100 g fish containing more than 0.2 ppm methoxychlor as shown by gas chromatographic analysis was extracted and cleaned by the method described above. Cleanup extract was concentrated to about 100 |.il, and 5- to 25-|xl samples were injected into the GLC/MS system. Recorded spectra were compared with those of reference methoxychlor. Presence of methoxychlor in water was also confirmed by GLC/MS. Results and Discussion RESIDUES IN RIVER WATER Methoxychlor was detected only in river water collected directly from treated water that passed the sampling point in Cecil Rapids (Table 1). No residues were detected in water collected either before or 1 week after the passage of the injected water. The leading edge of the treated water, indicated by drogues and the odor of the concentrate, required 2 hours to travel from the injection point to Cecil Rapids. Water collected at Cecil Rapids 1 5 minutes after arrival of the leading edge contained 0.16 ppm methoxy- chlor; water collected another 15 minutes later con- tained 0.14 ppm. The injected water thus required at least 30 minutes to pass this site, indicating a doubling of its volume during its travel 6.5 km downstream from the point of injection. At the same time methoxychlor concentration was reduced by 50 percent from 0.309 to 0.14-0,16 ppm. This indicates that the treated mass of water became progressively more diluted, simul- taneously expanding in volume as it moved down- stream to the point where sand and faunal samples were collected. The two water samples collected from Cecil Rapids 15 and 30 minutes after arrival of the leading edge contained about 85 and 125 ppm suspended solids, respectively. More than half the suspended solids con- sisted of very fine sand with particle diameters of 0.05- 0.1 mm. The remainder consisted of silt and clay particles and a small proportion of organic material. Suspended solids in these two water samples contained 892 and 437 ppm methoxychlor, respectively: filtrates contained 0.085 and 0.084 ppm. respectively. Thus the suspended solids contained about 47 and 40 percent, respectively, of the total methoxychlor extracted from the two water samples. Merna et al. (5) noted that when methoxychlor was added to lake water containing plankton, most of the compound eventually became associated with the par- ticulate fraction. Fredeen et al. (6) showed that sus- pended solids filtered from the South Saskatchewan River 68 miles downstream from the application point of DDT black fly larvicide contained 0.24 to 2.26 ppm DDT. The DDT absorbed onto these suspended parti- cles was believed able to act as a selective poison against filter-feeding insect larvae, including Simulium arcticum, the target species. Because methoxychlor is also readily adsorbed onto suspended particles, it could also be more toxic to filter-feeding insect larvae in the Saskatchewan River than to nonfilter feeders. Methoxy- chlor applied to the river did prove much more destruc- tive to Siiuiiliiim larvae than to Trichoptera larvae. However, nonfilter feeders such as Plecoptera larvae were also severely affected (2). RESIDUES IN SAND Nine days after methoxychlor injection, sand and silt collected from the riverbed 21-22 km downstream from the point of injection also contained methoxychlor (Table 1 ) . Sand from the north side of the river contained 0.1 ppm methoxychlor; sand from the south side contained 0.05 ppm. The difference in concentra- tions may have been caused by differences in sampling depths or by the varied proportions of sand and silt deposited on each side. Unfortunately no information is available on these characteristics. No residues were detected in samples collected before the May 23 in- jection. The limited number of samples did not enable authors to calculate the rate of methoxychlor loss from the injected water. The fact that no methoxychlor was detected in any pretreatment samples proves that resi- dues did not persist from treatments in 1971 or earlier. RESIDUES IN MUSSELS No methoxychlor residues were detected in mussels collected from riverbed sand 21-22 km downstream from the point of injection (Table 2). In 1968 Bedford et al. (7) showed that two species of freshwater mussels in the Red Cedar River in Michigan contained DDT and its metabolites, plus aldrin and methoxychlor. Concentrations of methoxychlor in these mussels range from nondetectable to 0.22 ppm. At the time of their study in the mid 1960s, methoxychlor, DDT, and mala- thion were used annually in the vicinity of the Red Cedar River to control elm bark beetles and mosquitoes. Thus it is presumed that mussels in the Red Cedar River were continuously exposed to these chemicals for long periods of time. Mussels collected from the river had been exposed to about 0.15 ppm methoxy- chlor in the water for only an hour or so; they were exposed to about 0.1 ppm in sand for about 8 days. Kapoor et al. (8) showed that in an aquarium ecosys- tem, snails exposed to methoxychlor for about 13 days accumulated relatively large amounts of the compound, whereas the methoxychlor content of fish remained in dynamic equilibrium with levels in their environment. RESIDUES IN INSECT LARVAE No methoxychlor residues were detected in the larvae of Odonata or Trichoptera collected alive from the river- bed either 6-7 days before or 8-9 days after treatment 244 Pesticides Monitoring Journal of the river with methoxychlor on May 23, 1972 (Table 2). However, larvae of these and other families of aquatic insects, including Plecoptera, Ephemeroptera, and Diptera, that were disabled by methoxychlor and collected with a net during the actual passage of injected water, contained an average of 17.5 ppm methoxychlor. Presumably fish that ate insects disabled or killed by such a treatment would be exposed to relatively high concentrations. RESIDUES IN PRETREATMENT FIS}) No methoxychlor was found in any pretreatment fish except goldeye (Table 3), although other chlorinated hydrocarbons were detected. DDT and related com- pounds were detected in 60 percent of the goldeye and 64 percent of the suckers; heptachlor and heptachlor epoxide were detected in 60 percent of the goldeye. Levels of these chemicals indicated by GLC were less than 0.1 ppm, too low to be confirmed by TLC. No residues of chlorinated hydrocarbons were detected in pickerel, sauger, or northern pike. RESIDUES IN POSTTREATMENT FISH Methoxychlor was detected in the muscle tissues of 66 percent of the goldeye collected 8-9 days after the 15-minute injection of 0.309 ppm methoxychlor about 21-22 km upstream from the fish-collecting site. In 37 percent of the goldeye, methoxychlor concentrations in muscle tissues ranged from 0.02 to 0.20 ppm; in 21 percent, concentrations ranged from 0.21 to 0.99 ppm; in 8 percent of this species, they ranged from 1.0 to 1.5 ppm; and in 34 percent, there were no detectable residues. Presence of methoxychlor could be confirmed in 47 percent of the goldeye by TLC, and in 21 percent by mass spectrometry. Mass spectra in these samples were the same as those of the reference methoxychlor. No methoxychlor was detected in any other species of fish collected 8-9 days after treatment. As in pretreatment samples, other chlorinated hydro- carbons were also detected in these fish. Residues were less than 0.1 ppm except in the goldeye where levels of up to 0.16 ppm aldrin and dieldrin and up to 0.51 ppm DDT and related compounds were detected. TABLE 2. Methoxychlor residues in clams and insect larvae collected from North Saskatchewan River, May -June — 1972 Date Methoxychlor Collected Location Specimen Content, ppm May 16 Lacolle Falls, N. side About 100 small mussels ND May 17 Lacolle Falls, N. side About 100 small mussels ND May 31 Lacolle Falls. N. side About 100 small mussels ND May 16 Lacolle Falls, N. side About 100 Odonata larvae ND May 17 Lacolle Falls, N. side About 100 Odonata larvae ND May 31 Lacolle Falls, N. side About 100 Odonata larvae ND June 1 Lacolle Falls, N. side About 30 Trichoptera larvae ND June 1 Lacolle Falls, S, side About 30 Trichoptera larvae ND May 23: before treatment Cecil Rapids, midriver About 50 insect larvae ND May 23: during treatment Cecil Rapids, midriver About 1,000 disabled insect larvae 17.5 NOTE: ND = nol detected «0.01 ppm methoxychlor). TABLE 3. Residues of chlorinated hydrocarbon insecticides in muscle tissues of fish collected from North Saskatchewan River before and after methoxychlor injection. May 1972 No. Fish Analyzed Heptachlor and Heptachlor Epoxide Aldrin and Dieldrin DDT, DDE, AND DDD Methoxychlor Common Name Percent OP Positive Sampli s Rancf, PPM Percent OF Positive Samples Range, ppm Percent OF Positive Samples Range, ppm Percent OF Positive Samples Range, ppm May 16 .tnd 17, 1972 Goldeye 25 60 0.02-0.05 40 0 III-0-08 60 0.01-0.04 0 ND Sucker 14 0 ND 0 ND 64 0.01-0.04 0 ND Pickerel and Sauger 12 0 ND 0 ND 1) ND 0 ND Northern Pike 8 0 ND 0 ND n ND 0 ND May 31 and June I. 1SI72 Goldeye Sucker Pickerel and Sauger Northern Pike 3 7 19 39 0 100 26 0.01-0,05 ND 0.02 0.02 29 (.6 14 0.01-0.16 0.02-0113 0.01 ND 0.03-0.51 0.03 ND 0.01 66 0 0 0 0.02-1.5 ND ND ND Scplcuihcr 19. 1972 Goldeye 6 0 ND 0 ND 0 ND 0 ND Sucker 9 0 ND 0 ND 0 ND 0 ND Pickerel and Sauger 6 0 ND 0 ND 0 ND 0 ND Northern Pike 2 0 ND {I ND 0 ND 0 ND NOTE: ND = not detected (<0.0I ppm). Vol. 8, No, 4, March 1975 245 RESIDUES IN FISH: 17 WEEKS POSTTREATMENT Methoxychlor could not be detected in the muscle tissues of any species of fish including goldeye col- lected from this same site September 19, 1972. RESIDUES IN FISH: GENERAL CONSIDERATIONS Considering their similar trophic levels, it was sur- prising that goldeye and other fish species had such disparate residues of methoxychlor 8-9 days after injection. The authors had expected relatively low concentrations of residues in goldeye, which graze mainly on insect colonies attached to rock surfaces, and relatively high concentrations in fish-feeding spe- cies such as pickerel, sauger. and northern pike, and in suckers, which are bottom-feeders and could have engorged on masses of immobilized larvae. The relatively higher concentrations of methoxychlor and other chlorinated hydrocarbons in goldeye muscle tissues may be explained by the relatively high oil content of their muscle tissues. Schmidt (9) reported that muscle tissue of one sample of six goldeye con- tained 6.57 percent fat; three samples of pickerel including 18 fish contained 2.84-3.34 percent fat; three samples of suckers including 18 fish contained 3.01-3.68 percent fat; and two samples of pike including 1 1 fish contained 1.40-1.86 percent fat. Let us assume that methoxychlor extracted from muscle tissues of goldeye fish in this study originated only from the fat fraction, and that these muscle tissues contained 6.57 percent fat. Then, in 37 percent of the goideyes, methoxychlor concentrations in fat fractions could have ranged from 0.3 to 3.0 ppm; in 21 percent, from 3.2 to 15.0 ppm; in 8 percent, from 15.2 to 22.7 ppm; and in 34 percent, concentrations would be less than 0.3 ppm. These levels do not demand govern- ment action, both because of the residues' ephemeral nature, presumably due to brief exposure in the river and rapid elimination from the fish, and because fish fat, unlike the fat of livestock, is not used separately as a food item for humans. In the case of livestock fat. concentrations of methoxychlor above 3 ppm rate government intervention. Conclusions Results indicate that a single 1 5-minute injection of 0.309 ppm methoxychlor into the Saskatchewan River contaminated water and biota for a short time at least as far as 22 km downstream. However, no detectable residues of methoxychlor remained from similar treat- ments of previous years. Like other organochlorine insecticides, notably DDT, methoxychlor was readily adsorbed onto suspended particles in water resulting in very high residue con- centrations. No residues were detected in insect larvae or mussels that inhabited the riverbed. Of all fish species examined here, only goldeye showed tendencies to accumulate methoxychlor. Concentrations which did accumulate in the edible flesh of about 80 percent of the goldeye were smaller than the concentration injected into the water 8-9 days earlier. The absence of methoxychlor residues in other species of fish may have been related to the relatively low oil content of their muscle tissues, as well as their ability to rapidly eliminate it from their systems as indicated by Kapoor et al. for Gambusia affinis fish (8). No methoxychlor residues were detected in goldeye or other fish 17 weeks after the experimental treatment. Acknowledgment Authors are indebted to Gordon Glen, Canada De- partment of Agriculture Research Station, Saskatoon, for assistance in collecting samples, and to R. H. Burrage and K. S. McKinlay, also of the Research Station, for assistance in preparing this manuscript. We are also indebted to N. Tomlinson, Assistant Director, Vancouver Laboratory. Fisheries Research Board of Canada, for permission to quote Mr. Schmidt's data from unpublished reports. LITERATURE CITED (/) Fretleen, F. J. H., J. G. Saha. and L. M. Rover. 1971. Residues of DDT, DDE and DDD in fish in the Sas- katchewan River after using DDT as a blackfly larvi- cide for twenty years. J. Fish. Res. Bd. Can. 28(1): 105-109. ^2) Fredeen, F. J. H. 1974. Tests with single injections of methoxychlor black fly (Dipleru: Simiiliidae) larvicides in large rivers. Can. Entomol. 106(3 ) :285-305. (.*) Sciha, J. G. 1971. Comparison of several methods for extracting chlordane residues from soil. J. Ass. Oflfic. Anal. Chem. 54( 1 ): 170-174. (4) Kovacs, M. F., Jr. 1963. Thin-layer chromatography for chlorinated pesticide residue analysis. J. Ass. Offic. Anal. Chem. 46( 5 ) :884-893. (5 ) i\teriw. J. W., M. E. Bender, and J. R. Novy. 1972. The effects of methoxychlor on fishes. I. Acute toxicity and breakdown studies. Trans. Amer. Fish. Soc. 101(2): 298-301. (6) Fredeen, F. J. H., A. P. Arnason, and B. Berck. 1953. .Adsorption of DDT on suspended solids in river water and its role in blackfly control. Nature (London) 171: 700. (7) Bedford. J. W .. E. W. Roelojs. and M. J. Zabik. 1968. The freshwater mussel as a biological monitor of pesti- cide concentrations in a lotic environment. Limnol. Oceanogr. 1 3( I ): 1 18-126. (S) Kapoor. I. P.. R. L. Melcalf. R. F. Nyslrom. and G. K. San!;ha. 1970. Comparative metabolism of methoxy- chlor. methiochlor and DDT in mouse, insects, and in a model ecosystem. J. Agr. Food Chem. 18(6) : 1145- 1152. (9) Schmidt. P. J. 1948. 1949. Analyses of freshwater fishes from Canadian interior provinces. Industr. Memo. F.R.B. Canada No. 9 and 12. Pacific Fish. Exp. St. Vancouver. Mimeo. 15 pp. 246 Pesticides Monitoring Journal Organochlorine Residues in Starlings, 1972 Paul R. Nickerson ' and Kyle R. Barbehenn ' ABSTRACT During the fall of 1972 starlings were collected from 130 sites in conjunction with the National Pesticide Monitoring Program. They were analyzed for DDT and its metabolites, dieldrin, heptachlor epoxide, benzene hexachloride. poly- chlorinated biphenyls and, for the first time in the series, oxychlordane and HCB. Mean DDT and dieldrin residue levels have declined significantly since 1967 and a regression analysis suggests that levels of DDT and its metabolites should fall below a mean of 0.1 ppm for the 1974 starling collection. Introduction As part of the National Pesticide Monitoring Program, a nationwide sample of starlings is collected and ana- lyzed biennially to help measure environmental levels of persistent organochlorine compounds. Data for resi- dues of these contaminants were first derived from an- alyses of three collections taken in 1967 and 1968 and reported by Martin in 1969 (/). The rationale for selecting starlings as a subject species appeared in that pilot study: they are a terrestrial avian species found year-round throughout most of the contiguous United States; they are generally regarded as expendable; and their omnivorous feeding habits can be expected to reflect pesticide intake from insects, fruits, grain, and miscellaneous other foods. Collections from fall 1 970 were reported by Martin and Nickerson in 1972 (2). The present paper presents data from the fall 1972 collection and compares portions of the data to those 1 Division of Technical Assistance, Fish and Wildlife Service, U.S. Department of the Interior, Washington, D.C. 20240. =* Criteria and Evaluation Division, Office of Pesticide Programs, Environmental Protection Agency, Washington, D.C. U.S. obtained in previous collections. Using these data, au- thors can examine some trends with greater confidence than in past studies and make tentative estimates for residue levels of DDT and dieldrin in future collections. Focus of the estimates will be national rather than re- gional, leaving the problem of substantial geographical variation for the future. Sampling As described in previous publications, the sampled area is composed of 40 blocks of 5° latitude and longitude, ranging from 24° to 49° latitude and 64° to 124° longi- tude {1,2). One to four starling sampling sites were randomly selected from each block. The same sites have been used for all collections. Figure 1 shows the location of all sampling sites and Table 1 lists locations successfully sampled during the November-December 1972 collection. Each sample location is identified by a row number, a column letter, and a site number: e.g., the site near Tacoma, Wash., is designated 1-A-l. In 1972, collections were made at 130 (94 percent) of 139 sampling locations. In previous years, Texas, a State of high pesticide use, posed a major problem. Thus collection efforts were intensified in 1972 and collections were successful in five of the nine preselected sites. Hopefully sampling in Texas will be improved further by extending the col- lection period into lanuary when the starlings are in larger flocks. Most starling samples consisted of a pool of 10 birds which had been trapped or shot. Each pool was wrapped in aluminum foil, placed in polyethylene bags, and frozen for shipment to the laboratory. Vol. 8, No. 4, March 1975 247 FIGURE 1. Starling monitoring sites, 1972 A nalytical Methods Residue analyses were performed as described previ- ously (.1,2) by WARF Institute, Inc., under contract with the U.S. Department of Interior — Bureau of Sport Fisheries and Wildlife (predecessor of the Fish and Wildlife Service). Technicians prepared the birds by removing their beaks and feet, then skinning their bod- ies. Each pool was ground in a Hobart food chopper until homogenized. The only major change in methodol- ogy involved use of 11 percent OV-17 -|- QF-1 (mixed phase) on 80/100 Gas-Chrom Q in the second column. Flow rate involved a retention time of 4-5 minutes for heptachlor epoxide. This new procedure separated oxy- chlordane from heptachlor epoxide, and HCB from BHC. For the determination of DDE, DDD, DDT, dieldrin, and polychlorinated biphenyls (PCB's), instrument con- ditions were: Temperatures: Column 200° C. Injector 230° C. Detector 240° C. Column: Carrier Gas: Flow: 4-ft-by-4-mm glass packed with 5 percent DC-200 on 80/100 Gas-Chrom Q. Nitrogen. Involved retention time of 6-8 minutes for p,p'- DDT. For the determination of HCB, alpha BHC, gamma BHC (lindane), beta BHC, heptachlor epoxide, and oxychlordane, instrument conditions were: Temperatures: Column: Carrier Gas: Flow: Column 180° C. Injector 220° C. Detector 235° C. 4-ft-by-4-mm 11 percent OV-17 + QF-1 (mixed phase) on 80/100 Gas-Chrom Q (available from Applied Science Cat. No. 12970). Nitrogen. Involved retention time of 4-5 minutes for hepta- chlor epoxide. Recovery rates for DDE, DDD, DDT, PCBs. dieldrin, and BHC ranged from 78 to 97 percent. Residue data were not corrected for recovery. Recovery and confir- matory tests were performed internally by the commer- cial laboratory on a quality control basis. Limits of detection ranged from 0.005 to 0.1 ppm. 248 Pesticides Monitoring Journal TABLE I. Starlini^ samplini^ sites listed by State and county, 1972 State Alabama Arizona Arkansas California Colorado Connecticut Florida Georgia Idaho Illinois Indiana Iowa Kansas Kentucky Louisiana Maine Maryland Michigan Minnesota Mississippi Missouri Montana County Marion Talladega Navajo Yavapai Maricopa Graham Yell /Pope Lonoke/Pulaski Colusa Shasta Modoc Ventura Stanislaus Monterey Inyo Kern Imperial Weld Montrose Crowley New London Bay Madison Polk Hardee Pike Wayne Nez Perce Owyhee Franklin Minidoka Stephenson Adams Kane Hendricks Fremont Jasper Marshall Rawlins Phillips Kearny Nemaha Marion Ohio Hopkins JeflFerson Rapides Penobscot Prince Georges Chippewa Grand Traverse Kent Ingham Aitkin Leake Harrison Jackson Butler Bollinger Meagher Blaine Missoula Riciiland Yellowstone Site Number State 3-H-l 4-H-3 3-C-3 3-C^ 4-C-l 4-C-2 3-G-2 3-G-3 2-A-l 2-A-2 2-A-3 3-A-l 3-A-2 3-A-3 3-B-l 3-B-4 4-B-l 2-D-4 3-D-l 3-D-2 2-D-2 4-H-l 4-1-3 5-1-1 5-1-2 4-H^ 4-1-2 1-B-l 2-B-l 2-C-3 2-C^ 2-G-l 2-G-3 2-H-2 2-H-3 2-F-3 2-G-2 2-G^ 2-E-l 2-E-2 3-E-l 2-F-4 3-F-2 3-H-2 3-H^ 4-G-3 4-G-4 l-K-2 2-J-l Nebraska New Mexico New York North Carolina North Dakota Ohio Oregon Pennsylvania South Carolina South Dakota Tennessee Texas Utah County l-H-I l-H-2 2-H-l Vermont 2-H^ Virginia I-G-4 4-G-l 4-G-2 Washingto 4-H-2 3-G-l 3-G^ Wisconsin 1-C-l l-C-2 Wyoming 1-C^ 1-D-l l-D-4 Keith Brown Lancaster Clay While Pine Humboldt Nye Clark Bernalillo Torrance Luna Olero Chaves Quay Jefferson Rensselaer Wilkes Union Macon Pender McLean Grand Forks Ransom Pickaway Wood Noble Greer Canadian Nowata Okmulgee Yamhill Lane Klamath Baker Harney Somerset Luzerne Aiken Potter Butte Hughes Brown Davidson Kinney Cochran Comal Clay San Patricio Weber Duchesne Sevier/Millard Grand Addison Amherst Prince George Caroline Pierce Yakima Spokane Whitman Clark Trempealeau Big Horn Crook Goshen Washakie Site Number 2-E-3 2-E-4 2-F-l 2-F-2 2-B-3 2-B-4 3-B-2 3-B-3 3-D-3 3-D-4 4-D-l 4-D-2 4-D-3 3-E-2 2-J-4 2nK-l 3-1-1 3-1-2 3-1-3 3-J-l l-E-3 1-F-l l-F-4 2-H 2-1-2 2-1-3 3-E-4 3-F-l 3-F-3 3-F-4 l-A-3 l-A-4 2-A-4 l-B-4 2-B-2 2-J-2 2-J-3 4-1-1 1-E-l l-E-2 l-E-4 l-F-3 3-H-3 4-E-3 4-E-4 4-F-l 4-F-3 5-F-l 2-C-l 2-C-2 3-C-l 3-C-2 1-K-l 3-1-4 3-J-2 3-J-3 1-A-l l-A-2 l-B-2 l-B-3 l-G-2 l-G-3 l-D-2 l-D-3 2-D-l 2-D-2 Vol. 8. No. 4. March 1975 249 Statistical Procedures National averages for DDT with its metabolites and dieldrin from ail five collections were calculated by two methods (Table 2). The geometric mean provides an appropriate measure of central tendency and should ap- proximate the median value in these skewed distribu- tions (see reference 2, Table 4). The arithmetic means average 2.2 times higher than the geometric means for DDT and metabolites, and 2.7 times higher for dieldrin. The arithmetic mean is a better measure of environ- mental load and is the appropriate value to use in deter- mining, for example, how much pesticide a Cooper's hawk would be likely to ingest if it ate 100 starlings. The estimate cannot be precise because of the low prob- ability of encountering starlings with unusually high concentrations of pesticides (Tables 3,4). In calculating the means, values given as <0.0I5-<0.010 ppm in the pilot study of this series (/) were assigned values of 0.005 ppm. For 1972. trace was called 0.002 ppm. The confidence intervals around the geometric means reflect the nationwide variability in residue levels and are not the most sensitive indicators of differences be- tween the annual means. Residue values of DDT and its metabolites from each station were compared with those of dieldrin for the periods summer-winter 1967-68 ver- sus fall 1968, fall 1968 versus fall 1970, and fall 1970 versus fall 1972. Statistically significant correlation co- efficients ranged from 0.59 to 0.70 for DDT and from 0.26 to 0.56 for dieldrin, indicating that a sign test (3) for paired values is an appropriate measure of temporal differences, even though the residual variance is high. Having established that certain differences between years were statistically significant, regression analysis TABLE 2. Geometric and arithmetic means of DDT and dieldrin residues in starlings, 1972 collectino Period No. Stations Sampled Lipids, % Wet Weight DDT & Metabolites, ppB Wet Weight Dieldrin, PPB Wet Weight Summer 1967 116 3.20 (3.01-3.41) 679 (536-861) AM 1755 27 (21-34) AM 82 Winter 1967-68 122 7,42 (6,99-7.87) 830 (670-1028) AM 2019 99 (74-134) AM 240 Fall 1968 122 5.84 (5.47-6.23) 569 (473-685) AM 1135 40 (32-49) AM 84 Fall 1970 125 5.40 (5.00-5.82) 445 (366-539) AM 916 36 (29^6) AM 117 Fall 1972 130 6.24 (5.89-6.61) 442 (420-464) AM 847 35 (28-45) AM 98 NOTE: Values in parentheses represent 95 percent confidence intervals for geometric i AM = Arithmetic mean. TABLE 3. Sites with residue levels of DDT and its meta- bolites greater than 3.0 ppm in baseline, 1970, or 1972 collections TABLE 4. Sites with residue levels of dieldrin greater than 0.3 ppm in baseline, 1970, and/or 1972 collections DDT Residues, ppm Site Number 1967-68 1970 1972 3-A-l 1.903 3.660 1.930 3-B^ 4.376 2,837 2.330 2-C-2 9.551 NS 0.640 3-C-2 3.163 NS 0.610 4-C-l 23.902 14.874 0.140 4-D-l 1.930 4,780 1.100 4-D-3 19.680 1.479 3.550 2-E-l 0.199 0,202 4.130 3-E-4 4.948 5.318 0.330 3-G-3 5.950 5.313 8.040 4-G-l 8.128 3.413 4.950 4-G-2 1.580 4.801 0.710 4-G-4 4.220 1.210 11.800 4-H-3 2.347 3,060 3.290 4-H-4 3.510 2,546 0.880 4-1-1 5.483 3.026 5.080 4-1-3 5.668 3.872 1.680 NOTE: NS = No sample collected. Dieldrin Residues, ppm Site Number Baseline Data, 1967-68 1970 1972 l-A-3 0.528 0.160 0.160 l-A-4 0.492 0.140 0.130 1-B-l 0.115 0.050 1.560 l-B-2 0.237 0.280 0.710 l-B-3 0.587 0.600 0.037 l-B-4 0.418 0.018 0.009 3-E-l 0.102 0.420 0.041 2-G-l 0.2B0 0.590 0.090 2-G-3 0.657 3.590 0.390 2-G-4 0.032 1.520 0.310 3-G-l 0.403 0.230 0.260 3-G-3 0.317 0.067 0.110 3-G-4 0.207 0.520 0.350 4-G-2 0.970 0.067 0.011 2-H-2 0.208 0.330 0.170 2-H-3 0.056 0.260 0.340 4-H-l 0.135 0.087 0.310 2-1-2 0.193 0.230 0.730 3-1-1 1.385 0.018 0.076 4-1-2 0.027 0.750 0.022 4-1-3 0.055 0.044 1.200 3-J-l 0.333 0.160 0.530 250 Pesticides Monitoring Journal was used to determine the relationship between residue levels and estimates of pesticide usage. The residue values for fall 1967 were estimated by converting the summer and winter values to a lipid basis and inter- polating on a logarithmic scale. Mean lipid values from the three fall collections (Table 2) were then used to estimate the wet-weight residue concentrations for the missing collection. Residue levels of organochlorine compounds from the 130 stations sampled in 1972 are presented in Table 5. Resulls and Discussion DDT AND METABOLITES Differences between the residue levels in collections from summer 1967 and winter 1967-68 were not statis- tically significant (Table 2). Levels in the fall 1968 samples, however, were consistently lower than those found in the corresponding averages for the summer/ winter samples, suggesting that a significant change in residue levels had occurred within what had been con- sidered the baseline jjeriods (/). Pesticide use preceding the four fall periods was assumed to be proportional to U.S. Department of Agriculture values for domestic disappearance (4). Residue levels and pesticide use are highly correlated (Fig. 2) and an extrapolation from the data points predicts a mean residue level well below 0.1 ppmfor 1974. Considerable caution should be exercised in projecting estimates far beyond this data base. The high correla- tion from only four data points could be fortuitous; authors believe use patterns to be more relevant than mere tonnage in determining exposure. Results from 1974 should provide a more reliable fix on the slope. Sites having DDT and metabolite residues greater than 3.0 ppm in baseline. 1970. and/or 1972 are listed in Table 3. Sites containing the highest DDT and meta- bolic levels in 1972 (greater than 3.0 ppm) were found in southeastern New Mexico (4-D-3), northwestern Kansas (2-E-l). central Arkansas (3-G-3), central Louisiana (4-D-3). north-central Alabama (4-H-3), and South Carolina (4-1-1 ). 3„0W / ^967 1968^/^ J)DT and Metabolites DDT y/^ Dieldrin logY=A+BlogX 1972-^^1970 X=1.138 X=1.422 ^ Y= 1.637 Y=2.763 / coef. of corr. coeff. of corr. ^ 0.969 / 0.8A1 r2=0.939 y A =0.879 / R^=0.708 . A=0.636 43 a B =1.325 • Antj.logs ^ 3=0.880 GO o X=26.4 million J^s. Antilogs ^ Y=579 ppb y ^ -2.0 X=13.7 million pounds o / 1967. Y=A3.0 ppb 03 >-< 4J / y^ c o u c: _ / ^^^Dieldrin Y / y^'^ o u / 1970^X4-1968 / y^\,ii m U c CO ■ / y X / ^ 1„0 / . ^ . . ^ . ( 5 1.0 2.0 Domestic disappearance of DDT and aldrin sales, log million of pounds. FIGURE 2. Regressions of pesticide residues in starlings on indices of pesticide use, 1967-68, 1970, 1972 Vol. 8, No. 4, March 1975 251 TABLE 5. Organochlorine residue levels in starlings, 1972 Site No.i Wet Weight, o Lipid Weight, c DDE DDD DDT DDT/ Meta- bolites PCB's = DiELDRIN Hepta- CHLOR Epoxide oxychlor- dane BHC HOB 1-A-l l-A-2 l-A-3 1-A-t 20.03 19.97 20.24 20.00 1.28 1.50 0.83 1.19 0.310 1.390 0.950 0.780 0.017 0.006 0.015 0.017 0.031 0.011 0.017 0.019 0.360 1.410 0.980 0.820 0.360 0.063 0.140 0.160 0.086 0.140 0.160 0.130 0.008 0.026 TR 0.006 0.007 TR TR TR 0.006 0.036 ND TR 0.007 3.330 0.036 0.011 2-A-l 2-A-2 2-A-3 2-A-4 20.16 20.08 20.02 20.09 0.53 0.85 1.69 1.13 0.470 0.630 0.077 0.490 0.008 TR TR 0.520 0.010 0.010 0.005 0.017 0.490 0.640 0.082 1.030 0.074 0.100 0.056 0.110 0.021 0.055 0.016 0.050 TR 0.009 TR 0.013 TR TR TR TR 0.011 TR 0.007 0.007 0.011 0.014 0.011 0.066 3-A-l 3-A-2 3-A-3 20.10 20.10 20.00 0.85 1.06 1.32 1.900 0.940 0.300 0.014 0.022 0.011 0.019 0.053 0.019 1.930 1.010 0.330 0.170 0.420 0.190 0.016 0.037 0.019 0.005 TR 0.006 0.008 0.005 0.006 TR TR 0.007 0.012 TR TR 1-B-l l-B-2 l-B-3 1-B^ 20.02 20.03 20.06 20.33 1.89 2.91 0.95 1.36 0.660 0.300 0.190 0.150 0.028 0.014 TR 0.008 0.021 0.021 0.01 1 0.012 0.710 0.330 0.200 0.170 0.120 0.170 0.037 0.100 1.560 0.710 0.037 0.009 0.009 0.010 0.210 TR TR 0.007 0.010 TR 0.006 TR 0.035 0.010 2.340 0.590 0.550 0.036 2-B-l 2-B-2 2-B-3 2-B-4 20.09 20.15 20.04 20.20 0.98 1.27 1.96 1.58 0.270 0.170 0.650 0.240 0.011 0.008 0.006 TR 0.029 0.017 0.014 0.006 0.310 0.200 0.670 0.250 0.230 0.120 0.250 0.099 0.060 0.019 0.046 0.019 0.007 TR 0.025 0.011 TR TR ND 0.006 0.006 0.009 0.009 0.006 0.057 0.006 0.007 0.009 3-B-l 3-B-2 3-B-3 3-B-4 20.02 20.29 20.05 20.01 0.62 0.91 1.56 1.39 2.340 0.180 0.620 2.310 0.009 TR 0.047 TR 0.034 0.006 0.160 0.022 2.380 0.190 0.830 2.330 0.370 0.099 1.870 0.140 0.070 0.039 0.270 0.037 0.017 TR 0.053 TR 0.017 0.009 0.031 0.006 0.005 0.006 0.009 TR 0.018 0.005 0.037 0.024 4-B-l 20.01 0.75 1.440 TR 0.017 1.460 0.190 0.026 TR TR TR 0.009 1-C-l l-C-2 l-C-4 20.54 20.05 20.48 1.26 1.28 1.05 0.091 0.078 0.081 0.009 0.016 0.013 0.032 0.019 0.024 0.130 0.110 0.120 0.370 0.210 0.290 0.025 0.021 TR 0.046 0.050 0.009 0.030 0.009 ND 0.010 0.016 0.005 0.006 0.007 0.009 2-C-l 2-C-23 2-C-3 2-C^ 20.18 20.24 20.11 20.14 1.39 1.13 1.02 1.66 0.350 0.630 0.530 0.970 TR TR 0.006 0.006 0.015 0.007 0.012 0.012 0.370 0.640 0.550 0.990 0.150 0.068 0.120 0.087 0.017 TR 0.017 0.028 0.008 0.008 0.027 0.021 0.006 TR 0.009 0.007 0.006 TR 0.005 0.006 TR TR TR 0.006 3-C-l 3-C-2' 3-C-3 3-C-4 20.00 20.16 20.02 19.98 1.66 1.60 0.99 1.01 0.630 0.590 0.590 0.490 TR 0.006 0.005 TR 0.009 0.011 0.020 0.014 0.640 0.610 0.610 0.500 0.150 0.200 0.250 0.110 0.034 0.160 0.012 0.300 0.011 0.012 TR 0.007 0.008 0.007 TR 0.006 0.006 0.006 TR 0.006 TR 0.006 TR TR 4-C-l 4-C-2 19.99 19.99 1.82 0.91 0.130 1.490 TR 0.011 0.009 0.022 0.140 1.520 0.081 0.160 0.021 0.020 0.008 0.006 TR TR 0.011 0.006 0.025 TR 1-D-l 1-D-2S l-D-3« l-D-4 20.34 20.09 20.02 20.38 1.39 1.02 1.66 1.85 0.025 0.077 0.058 0.076 TR TR 0.011 TR TR 0.006 0.052 0.009 0.025 0.083 0.120 0.085 0.037 0.100 0.750 0.061 0.018 0.005 0.009 0.025 0.005 0.006 0.007 0.007 TR TR TR TR 0.006 TR 0.006 0.005 TR TR 0.005 0.007 2-D-l 2-D-2' 2-V)-i 19.95 20.16 20.06 1.49 0.72 1.75 0.071 0.050 0.220 0.008 TR TR 0.017 0.006 0.016 0.096 0.056 0.240 0.250 0.099 0.190 0.006 0.006 0.049 0.010 0.009 0.011 ND ND 0.011 TR 0.007 0.007 0.005 TR TR 3-D-l 3-D-2 3-D-3 3-D^ 20.05 20.02 20.04 20.06 0.92 1.34 0.91 1.25 0.260 0.079 0.470 0.790 TR TR 0.017 0.005 0.012 0.009 0.042 0.014 0.270 0.088 0.530 0.800 0.120 0.094 0.450 0.160 0.066 0.007 0.007 0.021 0.013 TR TR TR 0.013 TR TR TR TR 0.009 TR 0.010 TR TR TR TR 4-D-l 4-D-2 4-D-3 19.99 19.90 19.99 0.85 0.61 1.26 1.090 0.630 3.520 TR TR 0.013 0.013 0.009 0.019 1.100 0.640 3.550 0.160 0.130 0.190 0.010 0.005 0.028 TR TR 0.019 TR TR 0.008 TR TR TR 0.005 TR 0.007 1-E-l l-E-2 l-E-3 l-E-4 20.01 20.11 20.12 20.13 2.00 1.60 0.95 2.10 0.120 0.068 0.078 0.023 0.009 0.008 0.005 0.008 0.022 0.017 0.007 0.027 0.150 0.093 0.085 0.058 0.120 0.190 0.099 0.220 TR 0.007 0.009 0.007 0.005 0.005 0.005 0.006 ND ND TR ND TR 0.006 TR 0.009 TR TR TR TR 2-E-l 2-E-2 2-E-3 2-B-4 19.98 20.01 20.16 20.13 0.87 1.42 1.26 1.16 4.070 0.190 0.110 0.056 0.023 0.014 TR 0.005 0.040 0.036 0.006 0.012 4.130 0.240 0.120 0.073 0.360 0.270 0.081 0.160 0.041 0.005 0.005 0.024 0.019 0.017 TR TR 0.005 0.005 0.011 TR 0.008 0.016 0.006 0.007 TR 0.007 TR TR 3-E-l 3-E-2 3-E^ 20.03 20.02 20.03 1.62 0.95 0.93 0.120 0.150 0.320 TR 0.014 TR 0.005 0.059 0.007 0.120 0.220 0.330 0.062 0.520 0.094 0.041 0.017 O.IOO 0.009 0.019 0.038 TR TR 0.009 0.007 0.006 0.007 0.009 0.027 0.006 4-E-3 4-E-4 20.14 20.23 1.23 1.03 2.060 1.020 0.006 TR 0.013 0.006 2.080 1.030 0.200 0.074 0.048 0.043 0.011 0.019 0.007 0.009 TR TR 0.007 0.009 1-F-l l-F-3 l-F-4 20.09 20.08 20.28 0.51 1.60 2.14 0.270 0.032 0.044 TR 0.005 TR 0.007 0.012 0.005 0.280 0.049 0.049 0.100 0.160 0.068 0.005 0.008 TR 0.006 0.006 TR TR TR TR TR TR 0.007 TR TR TR 2-F-l 2-F-2 2-F-3 2-F-4 20.21 20.21 20.09 20.04 1.86 1.77 1.33 1.41 0.330 0.230 0.170 0.150 0.027 TR TR TR 0.046 0.007 0.012 0.012 0.400 0.240 0.180 0.160 0.520 0.099 0.110 0.094 0.060 0.074 0.260 0.024 0.027 0.020 0.034 0.009 0.013 0.012 0.007 TR 0.009 0.012 TR 0.011 TR 0.005 TR 0.006 252 Pesticides Monitoring Journal TABLE 5 '(cont'd). Organochlorine residue levels in starlings, 1972 Site No." Wet Weight, G Lipid Weight, g DDE ODD DDT DDT/ META- BOLITES PCB's 2 DlELDRlN Hepta- CHLOR Epoxide oxychlor- dane BHC HCB 3-F-l 3-F-2 3-F-3 3-F-4 20.52 19.97 20.03 20.12 1.21 1.19 1.54 2.14 0.130 0.420 0.370 0.440 TR TR 0.032 TR 0.014 0.012 0.080 0.019 0.140 0.430 0.480 0.460 0.150 0.130 0.840 0.190 0.072 0.012 0.017 0.033 0.017 TR 0.008 0.007 0.007 TR 0.006 TR TR TR 0.012 TR 0.023 TR TR TR 4-F-P' 4-F-3 20.10 20. 1 3 1.09 1.10 1.060 0.620 TR TR 0.011 0.016 1.070 0.640 0.200 0.200 0.027 0.016 0.012 0.017 0.012 0.011 TR TR 0.025 TR 5-F-l 20.18 1.75 1.040 0.029 0.036 1.110 0.280 0.061 0.011 0.005 TR 0.008 l-G-2 I-G-3 l-G-4 20.15 20.11 20.28 1.26 2.11 1.67 0.230 0.390 0.150 0.008 0.023 0.017 0.022 0.053 0.035 0.260 0.470 0.200 0.220 0.550 0.320 0.027 0.016 0.020 0.009 0.017 0.020 TR 0.014 0.007 0.006 0.009 0.020 0.022 0.006 0.007 2-G-l 2-G-2 2-G-3 2-G-4 20.10 20.01 20.25 20.08 2.15 2.21 1.63 2.57 0.680 0.110 0.140 0.230 0.022 0.006 0.009 0.022 0.042 0.013 0.033 O.063 0.740 0.130 0.180 0.320 0.420 0.100 0.340 0.410 0.090 0.280 0.390 0.310 0.082 0.077 O.IIO 0.039 0.032 0.016 0.033 0.034 0.007 0.006 0.007 0.008 TR TR 0.019 TR 3-G-l 3-G-2 3-G-3 3-G-4 20.04 20.05 20.04 19.97 1.49 1.04 1.14 1.37 0.550 0.220 7.950 0.940 0.025 0.012 0.044 0.019 0.029 0.077 0.049 0.039 0.600 0.310 8.040 1 .000 0.210 0.720 0.220 0.290 0.260 0.035 0.110 0.350 0.034 0.057 ND 0.031 0.014 0.028 ND 0.018 0.006 0.008 TR 0.014 0.011 0.014 0.044 0.080 4-G-l 4-G-2 4-G-3 4-G-4 20.02 20.09 20.11 20.29 0.81 0.92 0.84 1.71 4.920 0.680 0.710 11.700 0.013 TR TR 0.089 0.020 0.028 0.091 0.046 4.950 0.710 0.800 11.800 0.160 0.290 1.020 0.220 0.039 0.011 0.016 0.150 0.021 0.012 ND 0.065 0.009 0.009 ND 0.018 TR TR TR 0.011 0.024 0.009 0.044 TR 1-H-l l-H-2 20.04 19.96 1.85 1.26 0.180 0.680 0.021 0.054 0.069 0.059 0.270 0.790 0.700 0.370 0.023 0.023 0.011 0.008 TR 0.008 0.007 0.005 0.007 TR 2-H-l 2-H-2 2-H-3 2-H^ 20.02 20.03 20.04 20.04 1.69 1.40 1.15 2.31 0.780 0.440 0.560 0.500 0.032 0.018 0.011 0.018 0.096 0.046 0.026 0.057 0.910 0.500 0.600 0.580 0.990 0.410 0.250 0.530 0.025 0.170 0.340 0.024 0.054 0.049 0.031 0.026 0.054 0.011 0.007 0.024 0.006 0.009 TR 0.024 0.006 TR 0.550 0.029 3-H-l 3-H-2 3-H-3 3-H-4 20.97 20.00 20.06 20.07 1.14 1.79 1.16 1.28 1.790 0.130 0.440 0.660 0.016 0.020 0.006 TR 0.030 0.049 0.031 0.01 1 1.840 0.200 0.480 0.670 0.170 0.520 0.370 0.110 0.065 ND 0.030 0.084 0.037 0.016 0.011 0.026 0.029 0.009 0.012 0.009 0.007 ND TR 0.008 TR 0.140 TR 0.095 4-H-l 4-H-2 4-H-l 4-H-4 20.07 19.98 22.04 20.11 0.91 1.03 1.06 0.86 1.060 0.780 0.890 0.870 TR 0.013 0.840 TR 0.022 0.028 1.560 0.013 1.080 0.820 3.290 0.880 0.290 0.300 19.900 0.250 0.310 0.025 0.029 0.280 0.091 0.053 0.071 0.028 0.100 0.025 ND TR TR ND TR TR 0.044 0.059 TR TR 2-1-1 2-1-2 2-1-3 19.98 20.05 20.02 1.75 1.79 1.44 0.180 0.500 0.260 0.041 0.046 0.009 0.036 0.065 0.031 0.260 0.610 0.300 0.320 0.650 0.250 0.170 0.730 0.028 0.041 0.150 0.024 0.021 0.034 0.010 ND 0.012 0.018 0.210 0.280 0.046 3-1-1 3-1-2 3-1-3 3-1-4 20.02 20.03 20.01 20.90 0.77 0.94 0.90 0.77 0.370 1.090 0.150 0.570 0.016 0.009 0.006 TR 0.042 0.040 0.016 0.011 0.430 1.140 0.170 0.580 0.470 0.440 0.160 0.160 0.076 0.160 0.028 0.031 0.023 0.011 0.008 0.020 0.025 0.009 0.006 0.008 TR TR TR TR TR TR TR 0.007 4-1-1 4-1-2 4-1-3 20.03 20.00 20.15 0.88 1.07 0.97 5.040 0.600 1.640 0.015 0.011 TR 0.024 0.053 0.037 5.080 0.660 1 .680 0.190 0.860 0.410 0.050 0.022 1.200 0.041 ND 0.014 0.012 ND 0.038 TR TR TR TR 0.010 TR 5-1-1 5-1-2 20.02 20.17 0,78 0.83 0.460 0.420 TR 0.013 0.020 0.029 0.4SO 0.460 0.190 0.320 0.033 0.017 ND TR ND 0.011 TR ND TR ND 2-J-l 2-J-2 2-J-3 2-J-4 20.08 20.12 20.09 20.09 0.45 1.36 1.43 2.02 0.380 0.310 0.380 0.150 0.008 0.007 0.007 0.007 0.021 0.019 0.023 0.016 0.410 0.340 0.410 0.1 70 0.230 0.170 0.210 0.200 0.1 20 0.047 0.068 (1.011 0.022 0.030 0.02S TR 0.015 0.021 0.012 TR TR 0.030 0.007 TR 0.042 0.035 TR TR 3-J-l 3-J-2 3-J-3 21.21 20.05 20.01 1.12 1.05 1.25 1.110 1.610 0.530 0.014 0.008 0.031 0.039 0.056 0.046 1.160 1.670 0.610 0.390 0.520 0.540 0.530 0.130 0.097 0.038 ND 0.021 0.035 ND 0.027 ND TR ND ND TR 0.014 1-K-l l-K-2 20.07 20.03 0.96 1.58 0.370 0.390 0.006 0.011 0.018 0.026 0.390 0.430 0.200 0.250 0.016 0.027 0.005 0.014 0.005 0.012 TR TR TR TR 2-K-l 2-K.2 20.28 20.22 1.11 1.17 0.890 0.250 0.007 0.012 0.052 0.026 0.950 0.290 0.960 0.290 0.006 0.021 0.007 0.019 TR 0.019 0.005 TR 0.008 0.007 NOTE: Weights expressed in grams; residues expressed in ppm (yg/g) wet weight. TR = trace. ND = not detected. ^ PCB levels estimated by examining peak between DDD and DDT with Aroclor 1254 as standard. 2 Samples from each site composed of 10 birds except as indicated. 3 Sample composed of 9 birds. * Sample composed of 7 birds, s Sample composed of 5 birds. •* Sample composed of 8 birds. Vol.. 8, No. 4, March 1975 253 DIELDRIN The difference between summer 1967 and winter 1967- 68 dieldrin residue levels was much greater than that for DDT, and this seasonal difference was highly sig- nificant (Table 2). Residue levels the following fall were significantly lower than the summer/winter means. The vast majority of dieldrin residues found in the en- vironment are derived from use of aldrin; aldrin/diel- drin sales (J) are roughly correlated with residue levels in starlings (Fig. 2). Results suggest that dieldrin levels may decline slightly in 1974 but should drop sharply in 1975 with the suspension of most uses of aldrin/ dieldrin. Again, a more reliable fix on the slope will be generated from future collections. Highest dieldrin residues (Table 4) were found in Washington (l-B-2), Idaho (1-B-l). Illinois (2-G-3), Iowa (2-G-4), Missouri (3-G-4), Indiana (2-H-3), Florida (4-H-l and 4-1-3), Ohio (2-1-2), and North Carolina (3-J-l). BHC The compound BHC, including alpha, beta, and gamma isomers, was found in nearly all samples taken. levels were extremely low; the highest was 0.036 ppm. BHC levels from the 1972 collection are not compared with those from past collections because HCB is being re- ported separately for the first time. HCB Although HCB has been used primarily outside the United States as a grain fungicide, it is possible that the compound has contaminated the environment in this Nation through industrial uses. In general, HCB levels were quite low, 0.1 ppm or less; almost half the samples had residues of 0.005 ppm or less. There were two sites which had high residue levels: l-A-2 in Washington (3.330 ppm) and 1-B-l in Idaho (2.340 ppm). Two other sites in Washington (l-B-2 and l-B-3) had HCB levels exceeding 0.5 ppm. HEPTACHLOR EPOXIDE Nearly all samples collected in 1972 contained residues of heptachlor epoxide. Levels were relatively low, rang- ing from 0.005 to 0.210 ppm; only three sites had levels exceeding 0.1 ppm. Oxychlordane, which has been re- ported with heptachlor epoxide in the past, is being re- ported separately this year. Direct comparisons between previous heptachlor epoxide levels and the 1972 results are not made at this time. OXYCHLORDANE A breakdown byproduct of chlordane, oxychlordane, was found in nearly all samples at very low levels. The highest residue detected was 0.1 ppm. PCB'S Except for three sites, PCB's were found in all samples at levels below 1 ppm. Exceptions were sites 3-B-3 in Nevada, 1.87 ppm; 4-G-3 in Louisiana, 1.020 ppm; and 4-H-3 in Alabama, 19.9 ppm. The 1970 Alabama level was estimated to be 24.3 ppm; further monitoring is planned at this site to attempt to identify the source of contamination. A cknowledgmen ts Collections were made by field personnel of the Bureau of Sport Fisheries and Wildlife. Regional Pesticides Specialists of the Divisions of Wildlife Services, Fishery Services, and Ecological Services were responsible for coordinating and reporting collections and assuring that samples were received in proper condition by the con- tract laboratory. These specialists are John W. Peterson, Boston, Mass.; Donald W. Hawthorne, Atlanta, Ga.; James B. Elder, Twin Cities, Minn.; Harry Kennedy. Albuquerque, N. Mex.; Robert H. Hillen, Denver, Colo.; and David J. Lenhart, Portland, Ore. Dr. Elder and William E. Martin, Ecological Services Division, Twin Cities, Minn., both provided valuable editorial assistance and I. C. T. Nisbet. Massachusetts Audubon Society, suggested the method for extrapolating seasonal estimates. LITERATURE CITED (/) Martin, William E. 1967. Organochlorine insecticide residues in starlings. Pestic. Monit. J. 3(2) : 102-1 14. (2) Marliii. William £., and Paul R. Nickerson. 1972. Organochlorine residues in starlings — 1970. Pestic. Monit. J. 6(l):33-40. (.?) Remington, Richard D., and M. Anthony Schork. 1970. Statistics with Applications to the Biological and Health Sciences. Prentice-Hall, Inc., Englewood Cliffs, N.J. 418 pp. (4) Fowler, D. Lee, and John N. Mohan. 1973. The Pesti- cide Review 1973. Agricultural Stabilization and Con- servation Service, USDA, Washington, D.C. 60 pp. (5) Sloan. J. 1973. Shell Ex. 111. Consolidated Aldrin/ Dieldrin Hearings, FIFRA Docket Numbers 145 et al., Washington, D.C. 254 Pesticides Monitoring Journal Organochlorine Residues in Alaskan Peregrines^ David B. Peakall,2 Tom J. Cade,2 Clayton M. White,3 and John R. Haugh < ABSTRACT Organochlorine residue levels in eggs of Alaskan peregrines have remained essentially constant over the period 1969-73 despite decreased usage of these compounds in the United States and Canada. Studies on reproductive success in Great Britain and data on eggshell-thinning suggest that DDE resi- dues above 20 ppm wet weight in peregrine eggs are asso- ciated with inability to maintain population levels. Residues in mainland Alaska are well above this critical figure and the reproductive rate is low. On the Colville River in north- western Alaska, the last young falcons will fledge in 1975 and the remaining adult population will disappear by 1980 unless the present rate of reproductive failure is drastically and quickly reversed. In the Aleutians, however, levels range from 5 to 7 ppm and the reproductive rate is adequate to maintain the population. Introduction The decline of the peregrine (Falco peregrinus) has been one of the most dramatic events ascribed to the use of persistent pesticides ( / ) . Although the species is too rare for specimens to be collected solely for analy- sis, a number of infertile or addled eggs and dead young have been obtained from Alaskan locales in re- cent years. This limitation on collection means that the data are not selected as randomly as researchers would 1 Supported in part by National Institutes of Healtli (DHEW) Grant 00306, State University of New York Research Foundation grants-in- aid, U.S. Atomic Energy Commission Contract AT (26-l)-I71, and Brigham Young University Research Division funds. ^ Ecology and Systemalics Section, Cornell University, Ithaca, N.Y. 14850 3 Zoology Department, Brigham Young University, Provo, Utah. * Biological Sciences Department, State University of New York, Binghamton, N.Y. like, a common problem with data on many top preda- tors. Following restrictions on uses of organochlorines instituted in North America in recent years, residue levels of DDT and its metabolites in lower trophic levels have decreased (2-5). Because it has been pre- dicted that organochlorine residues in higher trophic levels will continue to rise for several years after the input decreases (6). authors considered it important to examine levels in a top predator over this period. Alaskan peregrines can be divided into two main groups: interior and maritime. The mainland population is highly migratory and winters in Central and South America (/); the maritime population is essentially resident (7). Although both populations are essentially bird eaters, each group preys upon different species. White et al. (8) found that 65 percent, by weight, of the food of Aleutian peregrines consisted of alcidae, which winter offshore in the Aleutians. Residue levels of DDE ranged from 0.001 to 0.094 ppm wet weight for eight individuals of four alcid species (7). Truly migratory species comprised less than 3 percent of the prey. Cade et al. (9) found that migratory waterfowl com- prised half the food of the peregrines along the Yukon River; shorebirds and gulls represented about one quar- ter. Only one major prey species, the Canada jay (Perisoreus canadensis), is nonmigratory. In 1966 this species accounted for 1 1 percent by weight of the prey observed at aeries. A similar pattern was exhibited on the Colville River in 1967-69: of more than 45 species of prey found at aeries, only Ptarmigan (Lagopus lago- pus and L. mutus) are permanently resident in Alaska. The Ptarmigan comprise only 14 of 433 prey species {10). Vol. 8, No. 4, March 1975 255 Thus the mainland population not only migrates into areas of heavy pesticide use, but most of its prey species are also migratory. On the other hand, because Aleutian peregrines have little direct contact with pesticides, their levels reflect the background contamination of the North Pacific. Melliods In most cases eggs were blown by routine oological methods and the contents were immediately preserved in 10 percent formalin. If possible, eggs were rapidly shipped intact. Expression of residue levels on a wet- weight basis requires that all eggs be at the same stage of development and weighed shortly after collection. These conditions cannot be met for much of the field work in Alaska. Therefore, authors prefer to use the oven-dried weight. The amount of extractable fat was measured so that results could also be expressed on a lipid-weight basis. To compare results to those in much of the published work, divide ppm dried weight by 5 to give the wet-weight value. Samples were dried for 48 hours at 45 °C and then ground with coarse anhydrous Na2S04. Samples were extracted in Soxhlet thimbles for 8 hours with a 1:3 mixture of ethyl ether: petroleum ether. A florisil col- umn was used to remove fats and other interfering substances. A varian aerograph gas chromatograph equipped with "^Ni electron-capture detectors and two 0.6-cm-by-1.8-m columns was used for pesticide quanti- fication. Liquid phases and solid supports were 2 per- cent QF-1 on 40/50 Anakrom ABS and 5 percent SE- 30 on 60/80 Chromosorb W. Operating conditions were: Temperatures : Carrier Gas: Inlet 225° C, Detector 275° C. Column 200° C. Dry Nitrogen. Polychlorinated biphenyls (PCB's) were quantitated on two peaks after saponification. Recovery rates were 85 to 95 percent and values were not corrected. Results and Discussion Levels of DDE found in peregrine eggs during 1968-73 are listed in Table 1. There is no indication of any change in residue levels for either the interior or mari- time populations. Small sample sizes and the nonran- dom nature of the collections preclude statistical analy- sis. A few individual eggs were collected from the same site in different years, so comparisons can be made on a site basis. Two eggs from cliff 68 on the Colville River had DDE levels of 146 and 199 ppm oven-dried weight and shell index figures of 1.57 and 1.27 in 1968; an egg from this site in 1971 had a DDE level of 174 ppm and a shell index of 1.54. Shell index represents weight (mg) divided by the product of the length and breadth (mm) of the empty eggshell (//). In 1968 at Shivugak, on the Colville. the residue level was 276 ppm with an index of 1.57. In 1971 two eggs from this cliff had values of 215 and 352 ppm with index values of 1.46 and 1.37. Thus, neither from a consideration of individual sites nor from overall averages is there any evidence of a decrease in residue levels. Ten young peregrines were found dead in aeries along the Colville River in 1969; eight were found in 1971 (12). Nine of the 1969 young but none of the 1971 young could be analyzed. Residue levels in the muscle, liver, and brain are given in Table 2. These values are compared with those found in four young collected in 1966 (9). although they are not strictly similar. The 1966 specimens were alive when collected from various places, whereas the 1969 sample consisted of young found dead in the nest, apparently having starved after abandonment. Starvation may account for the higher residue levels in the brain in 1969. Nevertheless, even the highest brain levels, 42 ppm dry weight, are low compared to levels normally considered lethal (13). TABLE 1. DDE levels in Alaskan peregrine eggs, 1968-73 St. Dev. Shell Index Mean St. Dev. Thinning, ' Significance OF Thinning INTERIOR ALASKA Colville River 1968 11 193.9 159.8 11 1.42 0.13 24.9 <0.001 1969 163.8 87.4 5 1.53 0.15 23.5 <0.01 1971 210.8 92.0 7 1.49 0.09 21.2 <0.005 Yukon River 1968 11 105.6 53.0 11 1.49 0.10 16.8 <0.001 Tanana River 1969 344.3 81.0 3 1.42 0.14 20.2 ND 1973 302.7 84.1 3 1.44 0.18 19.1 ND ALEUTIANS Amchitka 1969 6 25.0 3.6 6 1.74 0.10 7.4 <0.01 1970 6 39.8 26.4 6 1.73 0.21 8.0 <0.05 1971 3 24.1 15.1 3 1.73 0.19 8.0 ND 1973 7 26.3 9.2 7 1.72 0.16 8.3 <0.05 NOTE: Pre-1946 values used to calculate percentage thinning for the various areas are: Colville 1.89, Aleutians 1.88, and Yukon 1.78; the value for the Yukon is probably too low (12). Data for the Colville 1968 and 1969, Yukon 1969, and Amchitka 1969 and 1970 have been reported previously (12) but not on a year-by-year basis. NDrzno data; sample loo small. 1 Residues are ppm oven-dry weight. 256 Pesticides Monitoring Journal TABLE 2. DDE levels in young peregrines found dead in aeries along Colville River, Alaska 19691 1966 2..1 Mean St. Dev. Range Mean Range Muscle Liver Brain 5.1 ± 2.0 11.7 ± 4.0 17.6 ± 4.7 1.2-19.0 1.2-33.0 1.1^2.0 6.3 1.6 1.6 0.8-14.6 1.3- 1.9 0.5- 3.0 NOTE: Residues are ppm oven-dry weight. ' Samples of all tissue represent nine individuals. 2 Samples of all tissue represent four individuals. ^ See Literature Cited, reference 9. The possibility that organochlorines were involved in the failure of parental care remains to be explored; abnormal behavior of adult falcons at their aeries cer- tainly appears to have increased in recent years. Three clutches of peregrine eggs laid in captivity during the Cornell breeding project have also been analyzed for residues. Eggs from a bird taken as a nestling from the Colville River in 1968 had DDE levels of 7.8 ppm dry weight and a shell index of 1.82 in 1972. Eggs from a Yukon River bird taken as a nestling in 1966 and raised on a diet of fresh fowl and coturnix quail had DDE residue levels of 9.0 ppm in 1972 and 7.7 ppm in 1973. Shell index figures were 1.79 and 1.80. respectively. The bird's controlled diet of fowl and quail thus resulted in low residue levels and a shell index approaching levels common before application. PCB levels follow quite a difTerent pattern from those of DDE (Table 3). In the Alaskan interior, the ratio of total DDT to PCB's is almost 1:1, whereas in the Aleutians the ratio is down to about 1:5. No other organochlorines were detected above trace amounts. TABLE 3. PCB levels in peregrine eggs, Alaska Year PCB 2DDT: Number Mean St. Dev. PCB INTERIOR ALASKA Colville River Tanana River 1971 1973 7 3 173.4 350.1 54.9 256.9 1.22 0.66 ALEUTIANS Amchitka Amchitka 1971 1973 3 7 114.4 144.6 77.4 68.3 0.20 0.18 NOTE: Residues are ppm oven-dry weight. EFFECT OF ORGANOCHLORINE RESIDUES The relation between DDE residues in eggs and the thickness of eggshells of Alaskan peregrines was plotted in 1971 (72). Data from the present study which are plotted in Figure 1 confirm the original plot. It is sig- nificant that residues from the lightly contaminated eggs of captive birds closely fit the regression line. A de- crease in thickness of 20 percent or more is associated with population declines (14); in the peregrine this critical thickness corresponds to about 20 ppm wet weight DDE in the eggs. Equivalents for dry weight and lipid are 100 and 500 ppm. respectively. Residues in eggs do not affect shell thickness, however; they are used merely as convenient indicators of circulating levels in mother birds at the time of egg maturation. Direct embryonic effects of DDE are probably unim- portant, because high levels are necessary to affect re- production in species in which eggshell thinning does not occur and when the eggs are incubated artificially (75,76). PCB's appear to be considerably more em- bryotoxic than DDE (17). It is not clear whether eggshell breakage is purely the result of structural failure, or whether it is caused by the mother bird's abnormal behavior on the nest. Rat- cliffe (18) feels that the behavioral component may be quite important. In the bird's diet. DDE levels of 40 ppm (79) and PCB's of 10 ppm (77) have been shown to cause behavioral changes leading to decreased repro- ductive success in ringed turtle doves (Streptopelia risoria). Ideally, one would like to determine what level of organochlorine residues in peregrine eggs interferes with the productivity necessary for a stable population. The best available data are from Great Britain. Ratcliffe showed that in 1962 reproduction was low in England, Wales, and southern Scotland, and that only in the Highlands of Scotland were peregrines reproducing nor- mally (18). In southern England only 3 percent of the sites had young who hatched. The rate in Wales was 4 percent; in nothern England, 7 percent, and in south- ern Scotland, 5 percent. In the Highlands of Scotland 38 percent of the sites had young who hatched. By 1971 substantial improvement had occurred in northern England and southern Scotland, btit reproduction was still poor in Wales and southern England. In 1971 the rate of sites hatching young was 7 percent in southern England, 4 percent in Wales, 24 percent in northern England, and 24 percent in southern Scotland. In the Highlands, this index of productivity remained essen- tially constant. Total organochlorine pesticide residue levels in eggs from northern England declined from 24.8 ppm wet weight in 1962-66 to 7.9 ppm in 1967-71. Corresponding values for southern Scotland were 14.5 and 11.0 ppm. Values for the Highlands were below 10 ppm throughout; no values are available for south- ern England or Wales for either period. About 90 per- cent of the residues are DDE. Nevertheless, other compounds, especially dieldrin, remain a problem. Lockie et al. noted that, concurrent with an increase of breeding success of the golden eagle (Aqtiila chrysae- tos). mean levels of dieldrin decreased from 0.86 to 0.34 ppm (20). This correlation does not apply to the Vol. 8, No. 4, March 1975 257 2.0 - • • 1.9 - • . o INTERIOR ALASKA • ALEUTIANS a CAPTIVE BIRDS 1.8 D • • X bJ 1.7 — V. Q • \s^ 2 _ \ ° ° o Li- o ° ^° °°oo ^ ?. 1.5 _ • \ ° CJ o o \>„^ 1- m \ < o O \. o o cr o \o 1.4 1 -X o o ° X„ \ . 3 o 1.2 1 o Ill c 10 20 30 40 60 80 100 200 300 400 DDE ppm (dried weight) FIGURE I. RcUilion belueen shell thickness index and DDE residues in eggs of Alaskan Peregrines. peregrine. RatclifTe's data show that the highest mean level of dieldrin. 1.2 ppm. occurred in the Highlands in 1962-66 when the peregrine was reproducing well {19). Residue levels in northern England decreased from 1962-66 to 1967-71. but they increased in south- ern Scotland. Although the residue levels represent 5-year periods, and the inlormation on reprodtictive performance rep- resents single years, and residiie levels are not related to reproduction at the specific aeries from which the eggs were collected, a tentative conclusion can be formed: namely, that the critical level of DDE in egg content is about 15 to 20 ppm wet weight. This value corresponds with the suggested critical value of 20 ppm DDE, wet weight, derived from consideration of the cggshell-lhinning data. These lines of evidence indi- cate that peregrines should continue to maintain normal population levels in the Aleutians where wet-weight means of egg content are 5 to 7 ppm. but not in the Alaskan interior populations where wet-weight means are 20 to 40 ppm. This hypothesis agrees with pub- lished observations of peregrines in the Aleutians (8) and interior Alaska (10.12,21) on reproductive suc- cess and population changes. As shown in Table 4, this relationship is particularly dramatic for peregrines nest- ing on the Colville River. If the present rate of repro- ductive failure is not drastically reversed soon, the last young falcons will fledge from the Colville aeries in 1975. and the remaining adult population will slowly disappear by 1980. The use of DDT in the United States has decreased almost linearly from a peak of 78 million pounds in 258 Pesticides Monitoring Journal 1959 to essentially zero in 1973 (22). Various moni- toring surveys have clearly indicated decreased levels of DDT and its metabolites over this period. Residue levels in estuarine moUusks surveyed from 1965 to 1972 showed a clear trend toward decreased levels of DDE beginning in 1969-70 (2). In 1970 Martin and Nickerson found that residue levels in starlings (Slums vulgaris) had decreased from 1967-68 levels at 35 of 38 sampling points (4); overall reduction of total DDT was 53 percent. Henderson et al. found that total DDT residues in fish had decreased at 33 of 50 sampling points from 1968 to 1969 (3). Ware et al., while study- ing the effect of a 2-year moratorium on the agricul- tural use of DDT in Arizona, found that levels in alfalfa and beef fat decreased significantly, but soil levels changed only negligibly (5). The rate of clearance indi- cated by these studies appears to be more rapid than had been predicted by Harrison et al. in 1970 (6); they estimated that the half-life of DDT for a given trophic level would be four times the average life span of the longest-lived species. TABLE 4. Number of breeding pairs and productivity of peregrines, Colville River, Alaska YEAR Total No. Pairs No. Pairs Producing Young i No. Young Produced i 1952-59 2 32-36 20-25 40-50 1967 3 27 18 34 1968' 32 16 34 1969 33 13 26 1971' 25 9 14 1973 14 4 9 1 Ranges are estimated. The entire river was not monitored every year. 2 Refer to Literature Cited, reference 26. =* Refer to Literature Cited, reference 12. * Refer to Literature Cited, reference 10. Samples of a few flickers (Colaptes auratus) killed by migrating falcons in Maryland and Virginia had residue levels of only a trace to a few tenths of a ppm. Flickers, which migrate the same time as the falcons, are an important food source for peregrines. The fact that levels of organochlorines in arctic peregrine eggs remain high suggests that these residues are obtained largely in Central and South American countries where their use has not been banned. Although the use of DDT in the United States, Canada, and many European counties has decreased consider- ably, there is no clear evidence that use is decreasing on a global scale. United States production of DDT remained between 100 and 180 million pounds from 1955 to 1970 and a steadily increasing proportion of the DDT was exported (22). Many countries in Cen- tral and South America, Africa, and Asia now manu- facture DDT. Monitoring levels of organochlorines in ocean plankton would trace the concentration of total DDT in the world's ultimate sink. Findings of the current study cause little optimism about the ultimate fate of North American peregrines. Residue levels remain high and reproduction low in mainland Alaskan peregrines. There is little to suggest that the 1970 forecast of extinction within a decade (21) was incorrect for some of the northern popula- tions, and the same bleak picture applies to the remain- ing Rocky Mountain population (23). The situation in Greenland, fortunately, appears more encouraging (24). The first major success in artificial breeding occurred in 1973 when 20 young peregrines were raised; two females from the Colville River mothered 13 of those 20 hatchlings (25). This was more than the entire natural productivity from the Colville River in 1973. Authors' findings suggest that reintroduction of captiv- ity-produced peregrines will be successful only if the released falcons do not migrate south of the United States. A cknowledgments Authors wish to thank the Alaska Department of Fish and Game for providing logistic support on the Col- ville River in 1973 and on the Tanana River in 1971-73. Thanks are also due F. Prescott Ward, who collected flickers in Maryland and Virginia for this study. LITERATURE CITED (/) Hickey, J. J., editor. 1969. The peregrine falcon popu- lations: their biology and decline. University of Wis- consin Press, Madison, Wise. (2) Butler, P. A. 1972. Organochlorine residues in estu- arine mollusks, 1965-72. Pestic. Monit. I. 6(4);238- 362. (3) Henderson, C, A. Inglis, and W. L. Johnson. 1971. Organochlorine insecticide residues in fish — fall 1969. Pestic. Monit. J. 5(1):1-11. (4) Martin, W. E., and P. R. Nickerson. 1972. Organo- chlorine residues in starlings — 1970. Pestic. Monit. J. 6{l):33-40. (5) Ware, G. W., B. J. Estesen, and W. P. Cahill. 1971. DDT moratorium in Arizona — agricultural residues after 2 years. Pestic. Monit. I. 5(3) :276-280. (6) Harrison, H. L., O. L. Loucks, J. W. Mitchell, D. F. Parkhurst, C. R. Tracy, D. G. Watts, and V. J. Yanna- cone, Jr. 1970. Systems studies of DDT transport. Science 170(3957) :5()3-508. (7) While, C. A/., W. B. Emison, and F. S. L. Williamson. 1971. Dynamics of raptor populations on Amchitka Island, Alaska. BioScience 21(12) :623-627. (S) White, C. M., W. B. Emison, and F. S. L. Williamson. 1973. DDE in a resident Aleutian Island peregrine population. Condor 75(3) : 306-3 11. (9) Cade, T. J., C. M. White, and J. R. Haugh. 1968. Peregrines and pesticides in Alaska. Condor 70(2): 170-178. (10) White, C. M., and T. J. Cade. 1971. Cliff-nesting rap- tors and ravens along the Colville River in arctic Alaska. Living Bird 10: 107-150. (//) Ratcliffe. D. A. 1967. Decrease in eggshell weight of certain birds of prey. Nature 215(5097) :208-210. (12) Cade, T. J., J. L. Lincer, C. M. White, D. G. Roseneau, and L. G. Swartz. 1971'. DDE residues and Vol. 8, No. 4, March 1975 259 eggshell changes in Alaskan falcons and hawks. Sci- ence 172(3986) :955-957. {IS) Stickel, L. F., W. H. Siickel, and R. Christensen. 1966. Residues of DDT in brains and bodies of birds that died on dosage and in survivors. Science 151(3717): 1549-1551. {14) Anderson. D. W ., and 1. 1. Hickey. 1972. Eggshell changes in certain North American birds. Proc. 15th Int. Ornithol. Congr. pp. 514-540. {15) Dewitt, J. B. 1956. Chronic toxicity to quail and pheasants of some chlorinated insecticides. J. Agr. Food Chem. 4(8) :863-866. {16) Lillie, R. J.. C. A. Denton, H. C. Cecil, ]. Bitman, and G. F. Fries. 1972. Effect of p,p'-DDT, o,p'-DDT and p,p'-DDE on the reproductive performance of caged white leghorns. Poultry Sci. 50(5 ): 1597-1598. (17) Peakalt, D. B., and M. L. Peakall. 1973. Effect of the polychlorinated biphenyl on the reproduction of arti- ficially and naturally incubated dove eggs. J. Appl. Ecol. 10(3):863-868. {IS) Ralcliffe, D. A. 1972. The peregrine population of Great Britain in 1971. Bird Study 19(3) : 1 17-156. {19) Haegele, M. A., and R. H. Hudson. 1973. DDE effects on reproduction of ring doves. Environ. PoUut. 4(1): 53-56. {20) Lockie, J. D., D. A. Ratcliffe, and R. Balharry. 1969. Breeding success and organo-chlorine residues in golden eagles in west Scotland. J. Appl. Ecol. 6(3): 381-389. (21) Cade, T. J., and R. Fyfe. 1970. The North American peregrine survey, 1970. Can. Field Nat. 84(3) :231- 245. (22) U.S. Department of Agriculture. 1972. Pesticide Re- view 1972. (23) Walker, W., W. G. Mattox, and R. W. Risebrough. 1973. Pollutant and shell thickness determinations of peregrine eggs from West Greenland. Arctic 26(3): 255-256. (24) Enderson, J. H., and J. Craig. 1974. Status of the peregrine falcon in the Rocky Mountains in 1973. Auk. 91(4):727-736. (25) Laboratory of Ornithology Newsletter. Summer 1973. No. 69. Cornell University, Ithaca, N.Y. (26) Cade, T. J. 1960. Ecology of the peregrine and gyr- falcon populations in Alaska. Univ. Calif. Publ. Zool. 63(2): 151-290. 260 Pesticides Monitoring Journal GENERAL Comparison Between Two Methods of Subsampling Blubber of Northern Fur Seals for Total DDT Plus PCB's ' Raymond E. Anas and Donald D. Worlund ABSTRACT Samples of 100 g blubber were collected from each of twelve 8- to 13-year-old fur seals (Callorhinus iirsinus) taken off the coast of Washington State in March 1972. Two methods of subsampling the blubber were compared. The mean level of total DDT (DDE, DDD, and DDT) plus polychlorinated biphenyls (PCB's) from a 5-g chunk of blubber taken from a 100-g sample was significantly less than the mean level from a 5-g subsample taken from the remainder of the blubber sample after it had been thorough- ly ground. Total DDT plus PCB residues ranged from 5.66 to 72.17 ppm, with a mean of 23.69 ppm in the chunks, and from 5.33 to 95.70 ppm, with a mean of 28.64 ppm in the homogenized blubber. Introduction Northern fur seals {Callorhinus ursimis) breed each summer on islands in the Bering and Okhotsk Seas, where they are harvested for their furs. Small breeding colonies are found on the Kuril Islands. Japan, and the Channel Islands off southern California. Fur seals are pelagic except during the breeding season. Their range extends from California to Japan. In the sampling of marine mammals for pesticide resi- dues the size of some organs and the blubber layer makes it necessary to take not only a relatively small sample from each animal, but also a subsample. Meth- ods of subsampling tissues from marine mammals for pesticide residues have varied widely. Holden and Mars- den (/) used 5-g aliquots of tissue from an original sample of unspecified weight. Anas and Wilson (2) and Aucamp et al. (i) collected and analyzed 10 g 1 Northwest Fisheries Center, National Marine Fisheries Service, Na- tional Oceanic and Atmospheric Administration, 2725 Montlake Boulevard East, Seattle, Wash. 98112. tissue; Arndt (4) collected 15-100 g tissue and ana- lyzed 10-g subsamples. Wolman and Wilson (5) col- lected 100 g tissue and analyzed subsamples of un- specified weight. The present report compares total DDT (DDE. DDD. and DDT) plus PCB (polychlori- nated biphenyl) residues from two methods of sub- sampling northern fur seal blubber to see whether the subsampling method affects results. Analytical Methods Two methods of subsampling the blubber were used: removing and analyzing a random 5-g chunk of frozen blubber from a 100-g sample; and grinding the re- mainder of the 100-g sample while frozen, stirring thoroughly, and removing and analyzing a random 5-g subsample. The -100-g sample of blubber consisted of a cross section taken from the belly area near the mid- line and anterior to the mammaries of 12 adult female northern fur seals taken off the coast of Washington State in March 1972. Samples were kept frozen at —23° C. Ages of the seals, determined by counting lines of den- tine in longitudinally sectioned upper canine teeth (6,7), ranged from 8 to 13 years. Both parous and nonparous seals were included. Unpublished data by the authors on 51 adult female northern fur seals ranging from 8 to 13 years have shown that mean organochlorine resi- dues are not associated with age or parous condition (P>0.05), so pregnant and nonpregnant seals from 8 to 13 years were pooled for the current study. Organochlorine residues were determined by WARF Institute, Inc., Madison, Wise. Methodology has been described previously; total DDT and PCB's were sum- med to reduce possible errors (8). Vol. 8, No. 4, March 1975 261 Results and Conclusions Total DDT plus PCB's ranged from 5.66 to 72.17 ppm. with an average of 23.69 ppm in the chunks, and 5.33 to 95.70 ppm with an average of 28.64 ppm in the homogenized blubber (Table 1). A paired t-test indi- cated that the average total DDT plus PCB level for samples from homogenized blubber was significantly greater than the level for chunks (P <0.05). Compari- son of values in Table 1 , however, suggests that the difference in results between the two sampling methods increases with increasing residue levels. The average difference of 4.95 ppm, therefore, may not be strictly applicable to the range of residue levels observed in this sample of 12 individuals. TABLE 1. Total DDT plus PCB's in blubber from 12 adult female fur seals, Washington State, March 1972 Type OF Sample Difference Sample No. Chunks Homogenized Ratio 1 5.66 5.33 -0.33 1.08 2 8.40 11.79 -1-3.39 0.71 3 14.78 16.68 + 1.90 0.89 4 17.15 17.34 -f0.19 0.99 5 15.05 17.93 4-2.88 0.84 6 17.60 17.96 +0.36 0.98 7 20.90 25.27 +4.37 0.83 8 25.86 28.04 +2.18 0.92 9 26.55 28.96 +2.41 0.92 10 16.73 29.78 + 13.05 0.56 11 43.47 48.89 +5.42 0.89 12 72.17 95.70 +23.53 0.75 Average 23.69 28.64 +4.95 0.83 NOTE: Data expressed in ppm; mg/kg wet weight. Ages ranged from 8 to 12 years. Ratios of residue values (chunk; homogenized) in Ta- ble 1 do not exhibit a trend with increasing residue levels. This hypothesis was indirectly tested by com- puting a weighted regression of residue values from chunks on those from homogenized samples and com- paring the estimated intercept with zero. The variance about the line was assumed proportional to the residue level, so weights used in the regression were constructed from reciprocals of residue values for homogenized samples and adjusted to sum to the sample size of 12. The intercept and its standard error were estimated to be 1.60 and 3.98, respectively. Because the standard error of the intercept is nearly 2.5 times larger than the intercept itself, there is little reason to reject the hypothesis that the intercept is zero. Therefore it ap- pears that residue values from chunks are a constant fraction of those from homogenized samples and that a regression through the origin adequately describes the relationship between results from the two sampling methods. Again, assuming the variance proportional to the res- idue level, the slope of the above regression is best estimated from the ratio of the average for chunks to the average for homogenized samples This value, 0.83, is significantly less than 1 (P <0.05) and indicates, as did the paired t-test, that significantly different residue values were obtained from the two subsampling methods used in this experiment (9). Analyses of residue levels expressed on a fat basis rather than a wet-weight basis produced the same results. The reason for the differences in pesticide levels by the two methods is not known. Obviously, one or both of the methods was not random. Differences could be caused by uneven distribution of organochlorine com- pounds or lipids in the blubber, or unequal separation of these compounds during subsampling. It is known, for example, that in finback whales (Balaenoptera physalus). the percent lipids in three sections of blub- ber is highest in the outer section and lowest in the inner section (70). Whatever the reason for the differ- ences, this study demonstrates that careful considera- tion should be given to the way in which subsamples are taken for pesticide analyses. LITERATURE CITED (/) H olden, A. V., and K. Marsden. 1967. Organochlorine pesticides in seals and porpoises. Nature 216(5122): 1274-1276. (2) Anas, R. E., and A. J. Wilson, Jr. 1970. Organo- chlorine pesticides in nursing fur seal pups. Pestic. Monit. J. 4(3):114-116. (i) Aucamp, P. J., J. L. Henry, and G. H. Slander. 1971. Pesticide residues in South African marine animals. Mar. Pollut. Bull. 2(12) : 190-191. {4) Arndt, D. P. 1973. DDT and PCB levels in three Washington State harbor seal (Phoca vitiilina richardii) populations. M.S. Thesis, Univ. Washington, Seattle, Wash. 65 pp. (.5) Wolman, A. A., and A. J. Wilson, Jr. 1970. Occur- rence of pesticides in whales. Pestic. Monit. I. 4(1): 8-10. (6) Scheffer, V. B. 1950. Growth layers on the teeth of Pinnipedia as an indication of age. Science 112 (2907);309-311. (7) Fisciis. C. H., G. A. Baines, and F. Wilke. 1964. Pe- lagic fur seal investigations, Alaska waters, 1962. U.S. Fish Wild!. Serv. Spec. Sci. Rept. Fish. 475. 59 pp. (8) Anas, R. E. 1974. DDT plus PCB's in blubber of har- bor seals. Pestic. Monit. J. 8(1) : 12-14. (9) Cochran, W. C. 1953. Sampling techniques. John Wiley and Sons, Inc. New York. 330 pp. (10) Ackman, R. G., C. A. Eaton, and P. M. Jangaard. 1965. Lipids of the fin whale (Balaenoptera physalus) from North Adantic waters. Canad. J. Biochem. 43(9):1513-1520. .262 Pesticides Monitoring Journal Degradation of Parathion Applied to Peach Leaves Wray Winterlin,' J. Blair Bailey," Larry Langbehn,' and Charles Mourer' ABSTRACT Parathion was applied to peach trees in three different formulations 70 days before harvest. Leaf samples were taken periodically through the 70-day period and gas-liquid chromatographic analyses were conducted for dislodgable and penetrated residues. Analyses were also conducted for paraoxon and the s-ethyl isomer of parathion. Punched samples were compared to whole-leaf samples; generally residue levels for both types corresponded closely. A new ex- perimental formidation, encapsulated parathion, produced highest levels of total parathion throughout the 70-day study, but even this formulation resulted in low total resi- due levels around I ppm at time of harvest. Degradation of the s-ethyl isomer of parathion was generally very rapid in all formulations studied. Dislodgable residues of paraoxon may be significant in some formulations and should he in- cluded in parathion degradation studies. Much of the par- athion found on peach leaves throughout the growing sea- son was dislodgable residue, but this depended considerably on the formulation used. Introduction Recent reports (1-6) have shown that foliage and other plant parts may accept and retain deposits of pesticide residues in much greater quantities and for longer times than does fruit, and that foliage may be a very important factor in considering worker reentry into insecticide-treated orchards. Gunther et al. (7) observed that a freestone peach tree about 10 years old has a leaf-to-fruit surface area ratio approximating 53:1; the 1 Department of Environmental Toxicology, University of California, Davis, Calif. 95616 - Extension Service, U.S. Department of Agriculture, University of California, Berkeley, Calif. same ratio for a clingstone peach tree is 28:1. Because foliage appears to be the greatest source of exposure of potentially toxic insecticides to farm workers, authors of the present study aimed to ascertain leaf residue levels of parathion and its toxic degradation products. Formulations can have an appreciable effect on de- posit and penetration levels as well as toxicity of insec- ticides (3,8,9). This study compares residue levels of two commonly used formulations, emulsifiable concen- trate (EC) and wettabie powder (WP), and an experi- mental formulation, encapsulated parathion (Enc). Both dislodgable and penetrated residues were mea- sured periodically for 70 days following application of each formulation. EC and Enc. formulations were ap- plied July 10, 1971, and the WP formulation was ap- plied August 3, 1971. El-Rafai and Hopkins (10) reported that paraoxon and possibly the s-ethyl isomer of parathion accumu- lated on both grass and leaf surfaces and were principal metabolites within the plant. The first few days after treatment, according to these investigators, paraoxon accumulated faster on leaf surfaces than it did inter- nally. The adverse was true several days later, however: surface paraoxon degraded more rapidly. Both paraoxon and the s-ethyl isomer are stronger cholinesterase in- hibitors than parathion, which in accumulated amounts could pose a threat to workers through dislodgable deposits. Therefore, the study reported here includes both these products as well as their parent compound, parathion. Analyses were conducted on punches taken at or near the center of leaves and on the whole leaf. Results Vol. 8. No. 4, March 1975 263 were compared on a weight basis. This portion of the study was designed to determine whether punched grape leaves would be representative for expressing dislodg- able residue in ug/cm- without sacrificing accuracy to sampling. Volunteer workers were permitted to enter the fields a few days following parathion application; samples of blood and urine were taken before and after their en- trance into the fields. Because this study involves only residues on peach foliage, findings involving human sub- jects will be reported elsewhere (I J). Materials and Methods FIELD APPLICATION AND SAMPLING A 60-acre orchard of freestone peaches southwest of Modesto. Calif., was subdivided into three 20-acre plots. Parathion was applied to each plot in one of the three formulations mentioned above: WP, EC, or Enc. The latter formulation encloses parathion in a porous micro- plastic capsule. This permits parathion to be released more slowly, over a longer period of time, than are most conventional insecticide formulations. The formulations were applied with a ground sprayer at rates of 1 lb active parathion per acre for both the EC and Enc, and 2 lb per acre for WP, Parathion was applied at 400 gallon/ acre for each of the three 20-acre plots, which is a standard applica- tion for peaches in most counties of California, In all three applications, sulfur was also mixed with the for- mulations and applied at a rate of 20 lb active sulfur per acre. Each 20-acre plot was subdivided into four replicate plots. Ten trees near the center of each repli- cate were selected for sampling. Whole-leaf and 2.5 cm- leaf-punch samples were taken one day before application, the day of application, and 3. 7. 14. 21. 28. and 70 days afterward. Twenty leaves and punches were taken from each tree selected at random in a cir- cular fashion at approximately 18^ intervals, 5 feet above ground. Samples from each 10-tree plot were combined, labeled, and frozen immediately in dry ice. Samples were soon transferred to a walk-in freezer where they were kept frozen at subzero temperatures until analyzed. E.XTRACTION AND ANALYSIS Samples of punched leaves were weighed and the grams per sample were recorded with the total number of punches. Whole-leaf samples were handled on a weight basis only and no attempt was made to measure leaf area. Each sample was extracted for dislodgable residue and for remaining residue according to the method of Gunther (7). Using this procedure, 200 leaf punches were transferred to a heavy-walled 500-ml Erlenmeyer flask. To each flask was added 200 ml of a 1:25.000 dilution of Sur-Ten wetting agent, also called aerosol OT 75 (trimethyl laurel ammonium chlo- ride). Contents of the flask were shaken for 60 minutes on a Gyrotory shaker at 180 rpm. The aqueous solution was decanted into a 1 -liter separatory funnel and an- other 200 ml diluted wetting agent was added. The solution was shaken for 30 minutes. The second portion of the wash liquid was combined with the first portion and a final 25 ml of wash solution was added to the leaves and shaken for 5 seconds. All Sur-Ten wash solutions were combined and extracted for 30 seconds with four 200-ml portions of chloroform. After filtering through sodium sulfate the organic solvent extracts were combined and stored in the dark until analyzed. The remainder residue, that which had penetrated, was extracted by transferring leaf contents remaining from Sur-Ten washings to a Waring blender container with 200 ml chloroform. Forty g sodium sulfate was added and the contents were blended for 2 minutes. The solvent was filtered through Whatman No. 1 filter paper into a storage bottle containing 20 g sodium sul- fate. An additional 200 ml chloroform was added to the blender cup and the contents were blended for 30 seconds and filtered. The extraction was repeated for 30 seconds with 200 ml chloroform, filtered, and the combined chloroform extracts were stored in the dark until analyzed. Whole-leaf samples were handled as the punched sam- ples had been. A 50-g portion of leaves was washed twice with 450 ml Sur-Ten solution in a 1 -liter flask, then washed again with a 100-ml solution. The solvent extraction was carried out by transferring 100 ml Sur- Ten wash solution representing 5 g leaves into a 250-ml separatory funnel and extracting four times with 50 ml chloroform. The remaining residue was handled in the same manner as the punches except that a larger quan- tity of sodium sulfate was added to the blending opera- tion: 100 g instead of 20 g. To remove residues from fruit samples, four peaches were selected, weighed, and measured, taking the diam- eter of each peach through three mutually perpendicu- lar axes. Surface area was calculated by averaging the diameters and assuming the peach was a sphere. The four peaches were transferred to a 1 -gallon can, sealed with 500 ml Sur-Ten solution, and rolled for I hour. The wash solution was decanted and the peaches were given a final wash with 500 ml Sur-Ten solution for 30 minutes. Washes were combined and an aliquot equiva- lent to 50 g was extracted as described for leaf sam- ples. Remainder or penetrated residues were extracted bv chopping only the edible portion of the fruit in a Hobart food cutter. Fifty-g chopped peaches were trans- ferred into a blender containing 50-g sodium sulfate. Two hundred ml chloroform was added to the blender and the contents were blended for 2 minutes. The ex- traction process was then handled according to the pro- cedure discussed for remaining residue in leaf tissue. 264 Pesticides Monitoring Journal LABORATORY ANALYSIS Because leaf samples contained high levels of sulfur, which interferes with the analysis of parathion, para- oxon. and the s-ethyl isomer of parathion, the extract had to be cleaned for detection by gas-liquid chroma- tography (GLC). It was also desirable to develop a pro- cedure by which paraoxon and the s-ethyl isomer could be separated from parathion prior to gas chromatog- raphy. Most GLC columns do not satisfactorily separate these materials, particularly when parathion has con- siderably more residue than the other two products. As a result there is an overlapping of GLC peaks which prevents satisfactory quantitative results. The cleanup procedure involved column chromatog- raphy using florisil as the adsorbant. A glass column, 2 cm in diameter and 12 cm long with a 150-ml reser- voir, was packed with 8 g PR grade florisil and pre- washed with 50 ml benzene. An extract of the sample in chloroform was evaporated to dryness and redis- solved in 5 ml benzene. The sample was quantitatively transferred to the column and eluted with 50 ml ben- zene; the eluate caused an interfering sulfur response which was discarded. Eighty ml 5 percent ethyl ether by volume in benzene was transferred to the column; this fraction contained the parathion that was collected. The column was then eluted with 75 ml 8 percent ace- tone in benzene; this fraction contained paraoxon and the s-ethyl isomer of parathion. Depending on the quan- tity of residue present, the sample was evaporated to dryness and the volume was adjusted to facilitate gas chromatographic analysis. Analyses were carried out with a Varian Aerograph Model 204 gas chromatograph equipped with a cesium bromide thermionic detector. Two gas chromatographic columns satisfactorily separated the three products. One column was a 2-ft-by-y8-in.-OD pyrex glass column packed with 5 percent (w/w) Dexsil 300 on 80/100 mesh Gas Chrom Q; the other column was a 2-ft-by-V8-in.- OD pyrex glass column packed with 5 percent Apiezon L on Gas Chrom Q, 80/100 mesh. Both columns were operated at 225° C with a slightly higher injection and detector temperature. Gas flow conditions were 19 cc/ min for nitrogen, the carrier gas; 16 cc/min for hydro- gen; and 150 cc/min for air. Recovery studies for parathion, paraoxon, and the s-ethyl isomer of parathion were performed before and during the study. Samples of control leaves were taken from plots prior to application and were fortified on leaf tissues during extraction before addition of sur- factant solution. Samples of control peaches were taken at harvest from trees not sprayed with parathion. Two levels of fortification at 0.1 and 1.0 ppm were made for each chemical analyzed. All recovered residues were based on total residue: that found in the surfactant solution plus the remainder found in the chloroform extract. Results and Discussion The method employed to separate and detect para- thion, paraoxon, and the s-ethyl isomer of parathion on and in peach leaves and fruit is reproducible and can detect residues as low as 0.01, 0.02, and 0.02 ppm, respectively, for each of the three chemicals. Fortified control studies at the 0.1 and 1.0 ppm level ranged between 90 and 100 percent, 85 and 95 percent, and 70 and 85 percent, respectively, for parathion, paraoxon, and the s-ethyl isomer. None of the data reported were based on or corrected for percent recovery. Residues found were frequently confirmed by repeated analysis and by using both the Apiezon L and the Dexsil 300 columns. Further confirmation would have been desir- able through gas-liquid chroniatography/mass spec- trometry and other detection systems; however, such instrumentation was not available. Recent reports {5,7) have shown that dislodgable resi- due is the principal means by which agricultural work- ers are exposed to pesticides. There are two means of measuring this dislodgable portion of the pesticide: one uses a weight basis in which data are usually re- ported in ppm; the other involves surface area only and is expressed as |,ig/cm-. Figures 1-3 illustrate the difference between expressing data as ppm and as l^ig/cm^ X 10^2. Taking into consideration normal field and laboratory variability, there appears to be very little difference between the two measurements with peach leaves. This is likely a result of the uniform density of the peach leaves. Once such uniformity has been estab- lished, data can be expressed in either form so long as one remembers that ultimately all dislodgable residue data must be understood in terms of surface residue to which a worker may be exposed. Table 1 compares this relationship with a final standard deviation between in- dividual samplings for the ratio of ppm/^g cm^ X 10~^. For each formulation the standard deviation did not exceed 0.09. The most practical way of measuring the dislodgable residue on surface foliage is through a representative punched sample. On the other hand, the punched sam- ple is usually taken near the center of the leaf or fo- liage. This may not be representative of the total leaf surface because on high-volume applications some in- vestigators have observed that residues on peach leaves appear to have their greatest deposition on the periph- ery of the leaf, particularly the tip. Therefore this study involved both whole-leaf sampling and punch sampling. Figures 4-6 show total and dislodgable residues ex- pressed on a weight basis (ppm) of the whole-leaf ver- sus the punched samples for each of the three formula- VoL. 8, No. 4, March 1975 265 tions. In all three formulations total residues found were similar in both types of sampling; yet the dislodgable portion of the total residue appeared to differ depend- ing on whether punched or whole-leaf samples had been taken. It is difficult to understand why this difference exists, but current studies indicate that higher dislodg- able residues in punched samples are primarily due to the extraction procedure, which is slightly different from that of whole-leaf samples in the ratio of surfac- : y WETTABLE POWDER PARATHION 50 1 10 }\ 5 ■ 1- X Ui 5 , U3 Z3 ■ ^0 = ■ v;;]l^*-~-____^ppM . ■ ^g/cm^xio'^ ' 0 1 T 0 05 ■ 0 01 3 10 20 30 40 60 60 7 DAYS POSTTREATMENT FIGURE 1. Paraihion residues from punched peach leaf sattzples following application of wettable powder formula- tion at 2 Ih a. i. /acre tant to total crop material. Until this difference can be resolved it is recommended that punched samples be used for analysis of dislodgable residues. When dislodgable residues are discussed for the en- capsulated formulation, they do not indicate residue levels to which a worker may be exposed: the capsule minimizes actual dermal exposure of the worker to the pesticide while extending the residual life of the active ingredient. The extraction procedure in this study re- moved the microcapsule containing the insecticide and any other residue that might be considered dislodgable from the leaf surface. Once Ihe encapsulated product has been removed, the pesticide contained in the capsule 100 2 — 1 1 1 1 1 1 - EMULSIFIABLE CONCENTRATE PARATHION 50 - 10 i 5 \ X -} g 2 ' O WO 6 ■ (E ' ■ ^^^J\,,__^^PPM 0 1 0 05 r ^ g/c m^ X id ^^~""^~^:;;:~^^^ 0 01 — ..1 . . — 1 1 . .1 . -_ 30 40 50 DAYS POSTTREATMENT FIGURE 2. Paraihion residues from punched peach leaf samples following application of emulsifiable concentrate formulation at 1 lb a.i./acre is extracted with a soluable organic solvent and nearly all the active pesticide is retrieved. Hence the plastic capsule does not act as a barrier between the pesticide and the solvent. Formulations played a major role in comparing per- cents of dislodgable residue removed. The percentage of dislodgable residue in the EC formulation from punched samples ranged between 17 and 33 percent with an average of 25 percent for seven sampling peri- ods. The WP formulation gave a higher percent of dis- lodgable residue and ranged between 43 and 61 percent with an average of 47 percent for seven sampling periods. Residue levels throughout the growing season degraded as one might expect with EC and WP parathion. With Enc. parathion the degradation as expressed by the slope was flatter, as anticipated. The percentage of dislodgable parathion compared to the remaining resi- due was considerably higher: about two-thirds of the total residue was dislodgable. Again it should be empha- sized that with this particular formulation dislodgable residues include both capsulated and noncapsulated 266 Pesticides Monitoring Journal 1 ; ENCAPSULATED PARATHION 1 50 V 10 \ ^ 5 : ^^ • ^^~>^:::r::~--P^ «• 1 - U (j/cn\ XI? ~ ■ -=^ D G w 0,5 • [C ■ 0 1 7 0 05 ■ 0 01 I 1 1 1 1 1 10 20 30 40 60 60 7 0 DAYS POSTTREATMENT FIGURE 3. Parathion residues from punched peach leaf samples following application of encapsulated formulation at I Ih a. i. /acre forms. What this means in actual exposure through oral and dermal contact cannot be ascertained from this study; additional work is needed to determine whether the parathion is actually in an exposed form. Enc. par- athion resulted in punched-Ieaf residues of 1.12 ppm or 10 fig/cm- at time of harvest. Whole-leaf samples had residues of 0.76 ppm or 7 ug/cm^ on 70-day har- vest samples. Paraoxon and the s-ethyl isomer, two potentially toxic degradation products of parathion, were also included in the study. Punched samples were limited so all analyses conducted for these two products are re- ported for whole-leaf samples only. Figure 7 shows the degradation of paraoxon throughout the 70-day study for each of the three formulations. Analyses were also separated according to their total and dislodgable resi- 30 40 DAYS POSTTREATMENT FIGURE 4. Parathion residues on and in whole peach leaf samples followini' application of wettahle powder for- mulation at 2 Ih a.i.l acre TABLE 1. Parathion residues in punched peach leaf samples Days Wettable Powder EMULStFIABLE CONCENTRATE Encapsulated Parathion POST- ^C/CM= X 10-2 PPM „C/CM= X 10-2 PPM jjC/CM=X 10-2 PPM j^G/CM^X 10 = „G/CM2 X 10-2 ;iG/CM2 X 10-2 0 69.5 60.8 1.14 18.4 14.7 1.25 34.9 31.5 1.11 3 16. U 11.4 1.41 3.16 2,64 1.20 19.6 14.6 1.34 7 5.12 41.5 1.23 1,00 0,815 1.23 10.4 8.02 1.30 14 1.88 1.45 1.30 n.47 0.387 1.21 5,18 4.27 1.21 21 0.62 0.488 1.27 0.28 0.214 1.31 3.11 2.48 1.25 28 0.54 0.415 1.30 0.24 0.204 1.18 2.40 1.96 1.22 70 0.20 0.157 1.27 0.065 0.060 1.08 1.12 1.03 1.09 Average 1.27 1.21 1.22 Standard 0.0653 0.0852 deviation 0.0755 Vol. 8. No. 4, March 1975 267 — I 1 1 1 1 1 EMULSIFIABLE CONCENTRATE PARATHION 30 40 DAYS POSTTHEATMENT ; 1 1 1 1 r 1 ENCAPSULATED PARATHION 50 ^ 10 5 e □. Q UJ cr h \ '^N^ "■"■-w 0,6 - KEY -, 0 1 7 whole-ieat tissues O ODrslodgable residue. 0 06 L whole-leaf tissues A ATotal residue. punched samptes A ADislodgable residue, punched samples 0 01 IC 20 30 40 50 60 70 DAYS POSTTHEATMENT FIGURE 5. Piinilhion resiiliies on anil in whole peach leaf samples jollowinf; application of enuilsiftahle concen- trate foriniilation at I Ih a. i. /acre FIGURE 6. Parathion residues on and in peach leaves fol- lowing application of encapsulated formulation at I lb a.i.l acre dues. Paraoxon was lowest with the EC formulation and was below the detectable limit the second day of sampling. Samples sprayed with the WP and Enc. formulations contained considerably higher levels of paraoxon than the EC formulation; the rate of degra- dation was similar to parathion, particularly after the third post-application day. Paraoxon was barely detect- able (0.01 ppm) at harvesttime. The greater percentage of paraoxon residue was found in the dislodgable form. This could be attributed to cither the fact that paraoxon has greater polarity than parathion. which would make it more extractable in the aqueous solvent, or to the diflference in rate of surface degradation compared to metabolic degradation. application. With the WP formulation, the residue de- clined from 0.28 ppm to 0.05 ppm during this same period. Nearly all residue from the Enc. application was in the dislodgable form; .^O percent of the WP application was in dislodgable form. This study indi- cates that residues of the s-ethyl isomer were low enough to be exckided from future studies involving parathion on peach trees. All dislodgable and remainder residues on harvested fruit samples were less than 0.01 ppm parathion. Neither paraoxon nor the s-ethyl isomer could be found on fruit samples. The s-ethyl isomer of parathion was detected in only very small quantities the first few days after application of the WP and Enc. formulations and was not at all detectable after the EC applications. Using the Enc. formulation, the residue declined from 0.11 ppm the day of application to 0.04 ppm the third day following Avkiiowledgwent Authors gratefully acknowledge the assistance of War- ren Dow. who supplied the 60 acres of peaches on which this study was conducled. Our thanks go also to Chevron Chemical Company and the Penwalt Cor- poration, who supplied formulations used in this study. 268 Pesticides Monitoring Journal 100 1 1 1 1 1 \ PARAOXON 50 . 10 - 5 ' 6 a a UJ KEY □ 1 - • • Total residue. WP Ui O 0 Dislodgable residue WP (T ■f^^. • • Total residue, ENC 0 5 ^ O O Dislodgable residue. ENC •- • Total residue. EC 0 1 r ^ ?-■--- 0 05 i 1 1 0 01 10 20 30 40 50 60 7 0 DAYS POSTTREATMENT FIGURE 7. Paraoxon on and in peach leaves following parathion application LITERATURE CITED (!) Bailey, J. B., D. Mengle, and D. H. Flaherty. 1972. Pesticide residues on grape leaves evaluated for ad- verse effects on grape pickers as related to worker re- entry periods. Unpublished report. (2) Carman, G. E., W. E. Westlake, and F. A. Gunther. 1972. Potential residue problem associated with low volume sprays on citrus in California. Bull. Envirop. Contam. Toxcol. 8(l):38-45. {3} Gunther, F. A. 1969. Insecticide residues in California citrus fruits and products. Residue Rev. 28:1-119. (4) Kilgore, W. W ., N. Marei, and W . Winterlin. 1971. Parathion in plant tissues: new considerations. Notes from National Academy of Sciences symposium on degradation of organic molecules in the biosphere, San Francisco, pp. 291-312. (5) Westlake, W. E., F. A. Gunther, and G. E. Carman. 1973. Worker environment research. Dioxathion (Del- nav) residues on and in orange fruits and leaves, in dislodgable particulate matter, and in the soil beneath sprayed trees. Arch. Environ. Contam. Toxicol. 1(1) :60- 83. (6) fVinterlin, W., C. Moiirer, and J. B. Bailey. 1974. Deg- radation of four organophosphate insecticides in grape tissues. Pestic. Monit. I. 8(I):59-65. (7) Gunther, F. A., W. E. Westlake, J. H. Barkley, W. Winterlin, and L. Langbehn. 1973. Establishing dis- lodgable pesticide residues on leaf surfaces. Bull. En- viron. Contam. Toxicol. 9(4) :243-249. {8) Durham, W. F. 1967. The interaction of pesticides with other factors. Residue Rev. 18:21-103. (9) Hull, H. 1970. Leaf structure as related to absorption of pesticides and other compounds. Residue Rev. 31:1-151. (10) El-Rafai, A., and T. L. Hopkins. 1966. Parathion ab- sorption, translocation and conversion to paraoxon in bean plants. I. Agr. Food Chem. 14(6) :588-592. (11) Bailey, J. B. 1973. Pesticide residues on peach leaves related to worker reentry periods. 168th Nat. Am. Chem. Soc. Meeting, Los Angeles, Calif., April 3, 1973. Vol. 8, No. 4, March 1975 269 APPENDIX Chemical Names of Compounds Discussed in This Issue ALDRIN BHC (BENZENE HEXACHLORIDE) CHLORDANE DDD DDE DDT DIELDRIN ENDRIN ETHYLENETHIOUREA HCB HEPTACHLOR HEPTACHLOR EPOXIDE LINDANE MALATHION METHOXYCHLOR OXYCHLORDANE PCB'S (POLYCHLORINATED BIPHENYLS) TDE TOXAPHENE Not less than 95% of l,2,3,4,10,10-Hexachloro-l,4,4a,5,8,8a-hexahydro-l,4-endo-cio-5,8-dimethanonaphthaIene 1,2,3,4.5,6-HexachlorocycIohexane (mixture of isomers). Commercial product contains several isomers of which gamma is most active as an insecticide. l,2,3,4,5,6,7,8,8-Octachlor-2,3,3a,4,7,7a-hexahydro-4,7-methanoindane. The technical product is a mixture of several compounds including heptachlor, chlordene, and two isomer forms of chlordane. See TDE. Dichlorodiphenyl dichloro-ethylene (degradation product of DDT) Main component (p.p'-DDE): l.l-Dichloro-2,2 bis(p-chlorophenyl) ethylene o,p'-DDE: l.l-Dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl) ethylene Main component (p.p'-DDT): l,l.l-Trichloro-2,2-bis(p-chlorophenyl) ethane. Other isomers are possible and some are present in the commercial product. o,p'-DDT: [l,l,l-Trichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl) ethane] Not less than 85% of l,2,3,4,10.10-Hexachloro-6.7-epoxy-l,4.4a,5.6,7.8,8a-octahydro-l,4-<'ndo-fio-5,8-dimethano- naphthalene l,2,3,4,10.10-Hexachloro-6,7-epoxy-l,4,4a,5,6,7,8,8a-octahydro-l,4-*ndo-<'ndo-5,8-dimethanonaphthalene 2-Iimdazolidmethione Hexachlorobenzene l,4,5,6,7,8,8-Heptachloro-3a,4,7,7a-tetrahydro-4,7-ffndo-methanoindene 1,4,5,6,7,8,8-Heptachloro 2,3-epoxy-3a.4,7,7a-tetrahydro-4,7-methanoindane Gamma isomer of benzene hexachloride (1,2,3,4,5,6-hexachlorocyclohexane) of 99+% purity 5-.[I,2-Bis(ethoxycarbonyl)ethyl] 0.0-dimethyl phosphorodilhioate l,l,l-Trichloro-2,2-bis(p-methoxyphenyl) ethane l,2.4,5.6.7,8,8-Octachloro-l,2-epoxy-3a,4,7,7a-tetahydro-4,7-methanoindan Mixtures of chlorinated biphenyl compounds having various percentages of chloride l,i-Dichloro-2,2-bis(p-chlorophenyl) ethane. Technical TDE contains some o,p'-isomer also. Chlorinated camphene (67-69% chlorine). Product is a mixture of polychlorinated bicyclic terpenes with chlorinated camphenes predominating. 270 Pesticides Monitoring Journal ERRATUM PESTICI'DES MONITORING JOURNAL, Volume 7, Number 3/4, p. 139. In the paper "Levels of Mirex and Some Other Organochlorine Residues in Seafood from Atlantic and Gulf Coastal States," a quotation of P. A. Butler's article "Monitoring Pesticide Pollution" [Bio- Science, 19(10)], which stated that mirex was "one of the most abundant of the organochlorine pesticides found in shellfish off the Atlantic, Gulf, and Pacific coasts," was misleading. Butler actually stated, "Al- though each sample is screened for 10 or more pesti- cides, DDT (including its metabolites) is the only one commonly present. Dieldrin is next in frequency of oc- currence, followed by endrin, toxaphene, and mirex." Acknowledgment The Editorial Advisory Board wishes to thank the fol- lowing persons for their valuable assistance in review- ing papers submitted for publication in Volume 8. Number 1-4, of the Pesticides Monitoring Journal: U.S. DEPARTMENT OF AGRICULTURE Kenneth R. Hill A. J. Malanoski Ralph G. Nash Donald W. Woodham U.S. ENVIRONMENTAL PROTECTION AGENCY Warren R. Bontoyan Henry F. Enos Cynthia M. Hemdon Alfred J. Wilson U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE Food and Drug Administration Paul E. Comeliussen LaVeme R. Kamps Sidney Williams George Yip U.S. DEPARTMENT OF INTERIOR Gary H. Heinz Vol. 8, No. 4, March 1975 271 SUBJECT AND AUTHOR INDEXES Volume 8, June 1974 — March 1975 Preface Primary headings in the subject index consist of pesti- cide compounds, the media in which residues are moni- tored, and several concept headings, as follows: Pesticide Compounds (listed alphabetically by common name or trade name when there is no common name) Media and Concept Headings Air Degradation Experimental Design Factors Influencing Residues Food and Feed Household Items Humans Plants (other than those used for food and feed) Sediment Soil Water Wildlife Compound headings are also used as secondary headings under the primary media and concept headings and vice versa.* When a particular paper discusses five or more organochlorines, the compounds are grouped by class under the media or concept headings; in the primary headings, however, all compounds are listed individu- ally. The specific compounds or elements which have been grouped in various combinations by class for cer- tain papers are as follows: Organochlorines aldrin BHC/ lindane chlordane chlorobenzilate DDE DDMU DDT dicofol dieldrin endosulfan endrin heptachlor heptachlor epoxide methoxychlor mirex nonachlor oxychlordane perthane Strobane® TDE toxaphene In the author index, the names of both senior and junior authors appear alphabetically. Full citation is given, however, only under the senior author, with a reference to the senior author appearing under junior authors. •Note: With the exception of 8(2):69-97 and 8(2):1I0-124 in which no compounds are used as secondary headings. 272 Pesticides Monitoring Journal SUBJECT INDEX Air Experimental Design 2,4-D 8(3):213-215 Rural DDT 8(3): 184-201 dieldrin 8(3):184-201 Urban DCPA 8(1): 53-58 Alachlor Soil 8(2):69-97 Aldrin Factors Influencing Residues 8(l):23-32 Household Items 8(2): 140-141 Sediment 8(4): 241-246 Soil 8(2): 69-97 Water 8(l):23-32 8(4): 241-246 Wildlife 8(1): 23-32 8(1):37^3 8(2): 142-143 8(4): 235-240 8(4): 241-246 Amiben, see Chloramben Amitrole Soil 8(2): 69-97 Aroclor, see PCB's Arsenic Food and Feed 8(2): 110-124 Atrazine SoU 8(2): 69-97 Azinphosmethyl Food and Feed 8(2):59-65 SoU 8(2):69-97 Azodrin, see Monocrotopbos Barban Sou Benefin Sou B 8(2):69-75 8(2):69-97 BHC/Lindane Factors Influencing Residues 8(3): 180-183 8(3): 209-212 8(4):219-224 Food and Feed 8(1):8-11 8(2):110-124 8(3):180-183 Household Items 8(2): 140-141 8(3):209-212 Humans 8(l):l-7 8(3):209-212 8(4):219-224 Sediment 8(3): 202-208 SoU 8(2):69-97 8(3): 202-208 Water 8(3): 202-208 Wildlife 8(2): 142-143 8(3):202-208 8(4): 247-254 Bidrin, see Dicrotophos CDEA Sou 8(2): 69-97 Borax SoU 8(2):69-97 Bordeaux Mixture SoU 8(2):69-97 Botran, see Dichloran Bromides Food and Feed 8(2): 110-124 Buturon SoU 8(2):69-97 Butylate Soil Bux Sou 8(2):69-97 8(2):69-97 Cadmium Food and Feed 8(2): 110-124 Calcium Arsenate SoU 8(2):69-97 Captan Soil 8(2):69-97 Carbaryl Food and Feed 8(2): 110-124 SoU 8(2): 69-97 Carbofuran SoU 8(2):69-97 Carbophenothion SoU 8(2): 69-97 CDAA Sou 8(2):69-97 Ceresan L® SoU 8 (2): 69-97 Ceresan M'B), see Granosan Ceresan Red Sou 8(2):69-97 Chevron RE-3535® Sou 8(2):69-97 Chloramben Soil 8(2):69-97 Chlordane, see also Oxychlor- dane Factors Influencing Residues 8(3):209-212 Food and Feed 8(1):8-11 8(4): 235-240 Household Items 8(3): 209-212 Humans 8(3):209-212 Sediment 8(l):33-36 Soil 8(2):69-97 Water 8(l):33-36 Chlorobenzilate Soil 8(2):69-97 Chloroneb Sou 8(2):69-97 Chloroxuron SoU 8(2):69-97 Chlorpropham Food and Feed 8(2): 110-124 SoU 8(2):69-97 CIPC, see Chlorpropham Copper Oxide SoU 8(2): 69-97 Copper-8-quinolinolate Sou 8(2):69-97 Copper Sulfate Degradation 8(4):225-231 Plants 8(4):225-231 Sediment 8(4): 225-231 SoUs 8(2):69-97 Water 8(4):225-231 Wildlife 8(4):225-231 Cotoran®, see Fluometuron Vol. 8, No. 4, March 1975 273 D 2,4-D Air 8(3):2I3-215 Food and Feed 8(2):110-124 Sediment 8(3): 173-179 Soils 8(2):69-97 Water 8(3): 173-179 Wildlife 8(2): 69-97 Dacthal®, see DCPA Dalapon Sou 2,4-DB Soil 8(2):69-97 8(2):69-97 DCPA Air 8(l):53-58 Factors Influencing Residues 8(3):209-212 Food and Feed 8(2):110-124 Household Items 8(3):209-212 Humans 8(3):209-212 SoU 8(2):69-97 Water a(l):53-58 WUdlife 8(l):53-58 DDD, see TDE DDE, sec also DDT Factors Influencing Residues 8(3):209-212 8(4): 219-224 Food and Feed 8(1):8-11 8(2): 110-124 8(3): 180-183 Household Items 8(2): 140-141 8(3):209-212 Humans 8(l):l-7 8(3): 148-156 8(3):209-212 8(4):219-224 Sediment 8(l):33-36 8(4):241-246 Water 8(l):33-36 8(4):241-246 Wildlife 8(1): 15-22 8(l):37-43 8(2): 105-109 8(2): 142-143 8(3): 162-166 8(3): 167-172 8(4):241-246 8(4): 247-254 8(4): 255-260 DDMU Wildlife 8(1):37^3 DDT, see also DDE, TDE Air 8(3): 184-201 Degradation 8(2):98-101 Experimental Design 8(4): 261-262 Factors Influencing Residues 8(l):23-32 8(3):209-212 8(4):219-224 Food and Feed 8(1):8-11 8(2):98-101 8(2): 110-124 8(3): 180-183 8(3):184-201 8(4):235-240 Household Items 8(2): 140-141 8(3):209-212 Humans 8(l):l-7 8(3): 148-156 8(3):209-212 8(4):219-224 Plants 8(3): 184-201 Sediment 8(l):33-36 8(3): 184-201 8(4):241-246 Soil 8(2): 69-97 8(2):98-101 8(3): 184-201 Water 8(I):23-32 8(1): 33-36 8(3):184-201 8(4):241-246 Wildlife 8(1): 12-14 8(l):15-22 8(I):23-32 8(1):37^3 8(21:105-109 8(2): 142-143 8(3): 162-166 8(3):167-172 8(3): 184-201 8(41:235-240 8(4):241-246 8(4): 247-254 8(4):261-262 DEF Soil 8(2):69-97 Degradation Food and Feed DDT 8<2):98-101 Fruit Trees parathion 8(4): 263-269 General mirex 8(2): 135-139 Water copper sulfate 8(4):225-231 8(21:69-97 Demeton SoU 2,4-DEP Soil 8(2):69-97 Diazinon Food and Feed 8(2): 110-124 Soil 8(2):69-97 Dicamba Soil 8(2):69-97 Dichloran Food and Feed 8(2):110-124 Dichloropropane/Dichloropro- pene SoU 8(2):69-97 Dicblorprop Sou 8(2):69-97 Dicofol Food and Feed 8(2): 110-124 Sou 8(2):69-97 Dicrotophos Sou 8(2):69-97 Dieldrin Factors Influencing Residues 8(l):23-32 8(3):209-212 8(4):219-224 Food and Feed 8(1):8-11 8(2):110-124 8(4):235-240 Household Items 8(2):140-141 8(3):209-212 Humans 8(l):l-7 8(3): 148-156 8(3):209-2I2 8(4):219-224 Sediment 8(41:241-246 SoU 8(2): 69-97 Water 8(l):23-32 8(4): 241-246 WUdlife 8(l):15-22 8(l):23-32 8(l):37-t3 8(2): 142-143 8(3): 162-166 8(31:167-172 8(4):235-240 8(4):241-246 8(4): 247-254 Dimethoate Soil 8(2):69-97 Dinex Sou 8(2):69-97 Dinitrobutylpbenol, see Dinoseb Dinitrocresol, see DNOC Dinitrocyclohexylphenol, see Dinex Dinoseb Sou 8(2):69-97 Dioxatbion SoU 8(2):69-97 274 Pesticides Monitoring Journal Diphenamid Soil 8(2):69-97 Disulfoton Soil 8(2):69-97 Dithane lVI-45®, see Mancozeb Diuron Soil 8(2):69-97 DNOC (Dinitrocresol) Soil 8(2): 69-97 Dodine Soil DSMA Soil 8(2):69-97 8(2):69-97 E Endosulfan Food and Feed 8(2): 110-124 Soil 8(2):69-97 Endothall Soil 8(2): 69-97 Endrin Factors Influencing Residues 8(3): 209-212 Food and Feed 8(2):110-124 8(4): 235-240 Household Items 8(3):209-212 Humans 8(3):209-212 Soil 8(2):69-97 Wildlife 8(l):37-43 8(2): 142-143 8(4): 235-240 EPN Soil EPTC SoU 8(2):69-97 8(2):69-97 Ethion Food and Feed 8(2):59-65 8(2):110-124 Soil 8(2):69-97 Ethylene Dibromide Soil 8(2):69-97 Ethylenethiourea Food and Feed 8(4):232-234 Experimental Design Air 2,4-D 8(3):213-215 Wildlife DDT 8(4):261-262 PCB's 8(4): 261-262 Factors Influencing Residues Biological Magnification aldrin 8(l):23-32 DDT 8(l):23-32 dieldrin 8(l):23-32 Formulation parathiou 8(4): 263-269 Interactions BHC/lindane 8(3):180-183 Occupation DCPA 8(3):209-212 organochlorines 8(3):209-212 Sex organochlorines 8(4):219-224 Species, Strain, or Race organochlorines 8(4):219-224 Falonel?', see 2,4-DEP Fensulfothion Soil 8(2):69-97 Fenthion SoU Ferbam Soil 8(2):69-97 8(2):69-97 Fluometuron Soil 8(2):69-97 Folex Soil 8(2): 69-97 Food and Feed Animal Feed malathion 8(4):235-240 organochlorines 8(4):235-240 Cereals ethylenethiourea 8(4):232-234 Fruits and Vegetables ethylenethiourea 8(4):232-234 General DDT 8(3): 184-201 dieldrin 8(3): 184-201 Grain and Forage DDT 8(2):98-101 Grapes organophosphates 8(l):59-65 Meat, Fish, and Poultry organochlorines 8(1):8-11 8(3): 180-183 PCB's 8(1):8-11 Total Diet 8(2): 110-124 Furadan®, see Carbofuran Granosan Soil 8(2):69-97 H HCB Food and Feed 8(2):110-124 Wildlife 8(4): 247-254 Heptachlor/Heptachior Epoxide Factors Influencing Residues 8(3):209-212 Food and Feed 8(1):8-11 8(1):110-124 Household Items 8(3):209-212 Humans 8(l):l-7 8(3):209-212 Sediment 8(4):241-246 SoU 8(2):69-97 Water 8(4):241-246 Wildlife 8(l):15-22 8(1):37^3 8(2): 142-143 8(4):241-246 8 (4): 247-254 Hexachlorobenzene, see HCB Household Items Carpeting organochlorines 8(2): 140-141 Dust DCPA 8(3):209-212 organochlorines 8(3):209-212 Humans Adipose organochlorines 8(I):l-7 Blood DCPA 8(3):209-212 organochlorines 8(3):209-212 8(4):219-224 MiUc DDE 8(3):148-156 DDT 8(3): 148-156 dieldrin 8(3):148-156 Isopestox®, see Mipafox Lasso's, see Alachlor Lead Arsenate Sou 8(2):69-97 Lindane, see BHC/Lindane Linuron Soil 8(2):69-97 Vol. 8. No. 4. March 1975 275 M Malathion Food and Feed 8(2): 110-124 8(4):235-240 Soil 8(2):69-97 Maleic Hydrazide SoU 8(2):69-97 Mancozeb Soil 8(2):69-97 Maneb Soil MCPA Soil 8(2):69-97 8(2): 69-97 Mercury Food and Feed 8(2):110-124 Wildlife 8(1): 15-22 8(2): 102-104 8(4):235-240 Methoxychlor Factors Influencing Residues 8(3):209-212 Food and Feed 8(2):110-124 Household Hems 8(3):209-212 Humans 8(3):209-212 Sediment 8(4):241-246 SoU 8(2): 69-97 Water 8(4):241-246 Wildlife 8(2): 142-143 8(4):24l-246 Methyl Demeton Soil 8(2):69-97 Methylmereury Dicyandiamide Soil 8(2): 69-97 Methyl Parathion, see also Parathion Food and Feed 8(2):110-124 Soil 8(2):69-97 Methyl Trithion Soil 8(2):69-97 Mevlnphos Soil 8(2):69-97 Mipafox SoU 8(2):69-97 Mirex Plants 8(2):135-139 Sediment 8(2): 135-139 SoU 8(2):69-97 8(2): 135-139 Water 8(2): 135-139 276 Wildlife 8(1): 15-22 8(2): 125-130 8(2): 131-134 Monocrotophos Sou 8(2):69-97 Monuron SoU PCNB Sou 8(2):69-97 MSMA Soil 8(2):69-97 8(2): 69-97 N Naled Food and Feed 8(2): 59-65 Naptalaiu Soil 8(2):69-97 Nitralin Sou Nitrate SoU 8(2):69-97 8(2):69-97 Norea Soil 8(2):69-97 NPA, see Naptalam o Oxychlordane, see also Chlordane Wildlife 8(4):247-254 Paraquat Sou 8(2):69-97 Parathion, see also Methyl Parathion Degradation K(4):263-269 Factors Influencing Residues 8(4): 263-269 Food and Feed 8(2): 110-124 Soil 8(2):69-97 PCB's Experimental Design 8(4): 26 1-262 Food and Feed 8(1):8-11 8(2): 110-124 Plants 8(3): 157-161 Sediment 8(l):33-36 8(3):157-161 Water 8(l):33-36 8(3):157-161 WUdlife 8(1):12-14 8(l):15-22 8(1): 37-43 8(2): 105-109 8(2): 142-143 8(3): 157-161 8(4): 247-254 8(4):255-260 8(4):261-262 PCP Food and Feed 8(2):110-124 Pentachlorophenol, see PCP Perthane Food and Feed 8(2):110-124 Pesticide Sales and Usage Ontario DDT 8(3): 184-201 o-Phenylphenol Food and Feed 8(2): 110-124 Phorate Soil 8(2):69-97 Phosalone Food and Feed 8(2):59-65 8(2): 110-124 Picloram Soil 8(2):69-97 Plants (other than those used for food and feed) Aquatic copper sulfate 8(4):225-231 PCB's 8(3): 157-161 Grasses mirex 8(2): 135-139 toxaphene 8(1):44^9 Tobacco DDT 8(3): 184-201 dieldrin 8(3): 184-201 Trees (Forest) resmethrin 8(l):50-52 Promctryne Soil 8(2):69-97 Propachlor SoU 8(2):69-97 Propanil SoU 8(2):69-97 R Ramrod « , see Propachlor Resmethrin Plants 8(l):50-52 Water 8(l):50-52 Ronnel Food and Feed 8(2): 110-124 Sediment, see also Soil, Water Lakes and Ponds copper sulfate 8(4): 225-231 Pesticides Monitoring Journal 2,4-D 8(3): 173-179 mirex 8(2): 135-139 PCB's 8(3):157-161 Rivers and Streams BHC/lindane 8(3): 202-208 chlordane 8(l):33-36 DDE 8(l):33-36 DDT 8(l):33-36 8(3): 184-201 dieldrin 8(3): 184-201 organochlorlnes 8(4):241-246 PCB's 8(l):33-36 TDE 8(l):33-36 Silvex Sou Simazine Soil 8(2):69-97 8(2): 69-97 Sodium Chlorate Soil 8(2):69-97 Soil, see also Sediment Croplands 8(2):69-97 DDT 8(2):98-101 8(3): 184-201 dieldrin 8(3): 184-201 Forest BHC/lindane 8(3):202-208 Pasture mirex 8(2):135-139 Strobane Soil SulfUT Soil 8(2): 69-97 8(2):69-97 Sutan®, see Butylate T 2,4,5-T Soil TCA Soil 8(2):69-97 8(2):69-97 TDE (DDD) Factors Influencing Residues 8(3):2()9-212 8(4):219-224 Food and Feed 8(2):110-124 Household Items 8(3):209-212 Humans 8(l):l-7 8(3):209-212 8(4):219-224 Sediment 8(1): 33-36 8(4): 241-246 Water 8(l);33-36 8(4): 241-246 Wildlife 8(l):15-22 8(1) 37^3 8(2) 105-109 8(2) 142-143 8(3) 162-166 8(4) 241-246 8(4) 247-254 Terbacil Soil 8(2): 69-97 Tetradifon Soil 8(2): 69-97 Thiram Soil 8(2):69-97 Toxaphene Food and Feed 8(2):110-124 8(4):235-240 Plants 8(i):44^9 Sediment 8(l):44^9 SoU 8(2):69-97 Wildlife 8(l):44-49 8(4): 235-240 Trifluralin Soil 8(2): 69-97 V Veriiolate Soil 8(2):69-97 w Water, see also Sediment Estuaries and Marshes aldrin 8(l):23-32 DDT 8(l):23-32 dieldrin 8(l):23-32 Groundwater PCBs 8(3):157-161 Lakes and Ponds copper sulfate R(4): 225-231 2,4-D 8(3): 173-179 DDT 8(3): 184-201 dieldrin 8(31:184-201 mirex 8(2):135-139 PCB's 8(3):157-161 Rivers and Streams BHC/lindane 8(3):2O2-208 chlordane 8(!):33-36 DCPA 8(l):53-58 DDE 8(l):33-36 DDT 8(l):33-36 8(31:184-201 dieldrin 8(31:184-201 organochlorines 8(41:241-246 PCB's 8(l):33-36 8(31:157-161 resmethrin 8(1):50-52 TDE 8(1) : 33-36 Wildlife Aquatic PCB's 8(3):157-161 Birds DDE 8(4):255-260 mercury 8(l):13-22 8(2): 102-104 organochlorines 8(l):15-22 8(l):37-43 8(4): 247-254 PCB's 8(l):15-22 8(41:247-254 8(4):255-260 Fish copper sulfate 8(4):225-231 2.4-D 8(3):173-179 DCPA 8(l):53-58 DDE 8(2):105-109 DDT 8(2): 105-109 8(3): 184-201 dieldrin 8(3):184-201 mercury 8(4): 235-240 organochlorines 8(4): 235-240 8(4):241-246 PCB's 8(2): 105-109 TDE 8(21:105-109 toxaphene 8(i);44-49 General mirex 8(2): 125-130 Invertebrates mirex 8(21:131-134 Mammals BHC/lindane 8(31:202-208 Plankton/Algae aldrin 8(l):23-32 DDT 8(l):23-32 dieldrin 8(l):23-32 Seals DDT 8(1):12-14 8(4):261-262 PCB's 8(1):12-I4 8(4):261-262 Shellfish aldrin 8(l):23-32 DDE 8(3):162-166 8(3): 167-172 DDT 8(l):23-32 8(3):162-166 8(31:167-172 dieldrin 8(l):23-32 8(3):162-166 8(3): 167-172 organochlorines 8(4):261-262 TDE 8(31:162-166 toxaphene 8(1):44^9 Wolves organochlorines 8(2): 142-143 Vol. 8, No. 4. March 1975 277 AUTHOR INDEX Aldrich, F. D., see Starr, H. G., Jr. Anas, R. E. DDT plus PCB's in blubber of harbor seals. 8(1): 12-14 Anas, R. E., and Worlund, D. D. Comparison between two methods of subsampling blubber of northern fur seals for total DDT plus PCB's. 8(4):261-262 Anderson, J. W., see Petrocelli, S. R. Andrews, T. L. Resmethrin residues in foliage after aerial applica- tion. 8(1): 50-52 B Bailey, J. B., see Winterlin, W. Balba, M. H., see Fredeen, F. J. H. Barbehenn, K. R., see Nickerson, P. R. Baulu, p., see Pecka, Z. Berst, a. H., see Frank, R. Blanke, R. v., see Griffith, F. D., Jr. Braun. H. E., see Frank, R. Cade, T. J., see Peakall, D. B. Cahill, W. p., see Ware, G. W. Campbell, D.. see Mes, J. Carey, A. E., see Crockett, a. B. Choi, P. M. K., see Zitko, V. Clark. D. E., Smalley, H. E.. Crookshank, H. R , and Farr. F. M. Chlorinated hydrocarbon insecticide residues in the feed and car- casses of feedlot cattle, Texas— 1972. 8( .1 ): 1 80-183 Clark, D. R., JR , and McLane, M. A. R. Chlorinated hydrocarbon and mercury residues in woodcock in the United States, 1970-71. 8n):15-22 Clegg, D. E. Chlorinated hydrocarbon pesticide residues in oysters (Crassostrea commerc talis) in Moreton Bay, Queensland, Aus- tralia—1970-72. 8(.1):162-166 Coffin, D. E., see Mes, J. Collins, H. L., Markin, G. P., and Davis, J. Residue accumulation in selected vertebrates following a single aerial application of mirex bait, Louisiana — 1971-72. 8( 2 ): 125-1.10 Collins, H. L., see Markin, G. P. Corneliussen, p. E., see Manske, D. D. Cr\btree, D. G., see Reidinger, R. F , jR Crockett, A. B , Wiersma, G. B , Tai, H., Mitchell, W G., Sand, P.F., and Carey, A. E. Pesticide residue levels in soils and crops, FY-70 — National Soils Monitoring Program (II). 8(2):69-97 Crockett, a. B , Wiersma. G. B., Tai, H., and Mitchell. W. G. Pesticide and mercury residues in commercially grown catfish. 8(4): 2.15-240 Crookshank, H. R., see Clark, D E. Crump-Wiesner, H. J., Feltz, H. R., and Yates, M. L. A study of the distribution of polychlorinaled biphenyls in the aquatic en- vironment. 8(3): 157-161 D Davis, J., see Collins, H. L. Davis, J., see Markin, G. P. Day, N. E., see Wassermann. M. Durant, C. J., see Reimold, R, J. Estesen, B. J., see Ware, G. W. Farr, F. M., see Clark, D. E. Feltz, H. R., see Crump-Wiesner. H. J. Fisher. F. M., Jr . see Ginn, T. M. Frank, R., Montgomery, K., Braun, H. E., Berst, a. H., and Loftus, K. DDT and dieldrin in watersheds draining the tobacco belt of southern Ontario. 8(3): 184-201 Fredeen, F. J. H., Saha, J. G., and Balba, M. H. Residues of melhoxychlor and other chlorinated hydrocarbons in water, sand. and selected fauna following injections of melhoxychlor black fly larvicide into the Saskatchewan River, 1972, 8<4):241-246 GiNN, T. M., and Fisher, F. M., Jr. Studies on the distribution and flux of pesticides in waterways associated with a ricefield — marshland ecosystem. 8(l):23-32 Goerlitz, D, F,, see Law, L. M. Gomes, E. D., see Miller, F. M. Graca, I.. SiLVA Fernandes, a. M. S., and Mourao, H. C. Organo- chlorine insecticide residues in human milk in Portugal. 8(3);148- 156 Greichus, Y. a., see Schneeweis, J. C. Griffith, F. D., Jr., and Blanke, R. V. Blood organochlorine pesti- cide levels in Virginia residents. 8(4):219-224 Groner, Y., see Wassermann, M. Grover, R., and McCashin. B. A nomograph for the conversion of 2.4-D ester concentrations in air from ^g/m^* to ppby and vice versa. 8(31:213-215 H Hanks, A. R., see Petrocelli, S. R. Harman, P. D., see Schultz, D. P. Harvey, E. J., Sr.. see Knight. L. A., Haugh. J. R , see Peakall, D. B. Hetzler, H., see Mick, D. L. Jr. Jackson, M. D-, Sheets, T. J., and Moffett, C. L. Persistence and movement of BHC in a watershed. Mount Mitchell State Park, North Carolina— 1967-72. 8(31:202-208 K Knight. L. A.. Jr.. and Harvey. E. J.. Sr. Mercury residues in the common pigeon {Cohtmha U\ta) from the Jackson, Mississippi, area— 1972. 8{2):102-104 Lanobehn, L., see Winterlin, W. Law, L. M., and Goerlitz, D. F. Selected chlorinated hydrocarbons in bollom material from streams tributary to San Francisco Bay. 8(1):33-16 Lazarovici, S., see Wassermann, M. LiNDER, R. L.. see Schneeweis, J. C. Lister, N. A., see Zitko, V. Loftus, K., see Frank. R. M Manske. D. D., and Corneliussen. P. E. Pesticide residues in total diet samples (VII). 8(2):110-I24 Markin. G. P. Collins, H. L., and Davis, J. Residues of the in- secticide mirex in terrestrial and aquatic invertebrates following a single aerial application of mirex bait, Louisiana — 1971-72. 8(2): 131-134 Markin, G. P., see Collins, H. L. Markin, G. P., see Spence, J. H. McCashin, B.. see Grover. R. McDougall. W D . III. see Starr. H G.. Jr. McIntosh. A. W. Degradation of copper in ponds. 8(4):225-231 McLane. M. A. R , see Clark, D. R. Mes. J.. Coifin. D. E.. and Campbell, D. Polychlorinaled biphenyl and organochlorine pesticide residues in Canadian chicken eggs 8(11:8-11 Mick, D. L., Hetzler, H , and Slach, E. Organochlorine insecticide residues in carpeting. 8(2): 140-141 Miller, F. M.. and Gomes, E. D. Detection of DCPA residues in environmental samples. 8(l):53-58 Mitchell. W. G.. see Crockett, A. B. Moffett, C. L., see Jackson, M. D. Monachan. D. J., see Zitko. V. Montgomery. K.. see Frank. R. MouNcE. L. M., see Starr. H. G., Jr. Mourao, H. C, see Graca, I. MouRER, C, see Winterlin. W. 278 Pesticides Monitoring Journal N Newsome, H., see Pecka, Z. NiCKERSON, p. R., and Barbehenn, K. R. Organochlorine residues in starlings, 1972. 8(4):247-254 Spence, J. H., and Markin, G. P. Mirex residues in the physical environment following a single bait application, 1971-72. 8(2);135- 139 Starr, H. G., Jr., Aldrich, F. D., McDougall, W. D., Ill, and MouNCE, L. M. Household dust as an index of human exposure to pesticides. 8(3):209-212 PE4KALL, D. B., Cade, T. J., Whiie, C. M., and Hauch, J. R. Organo- chlorine residues in Alaskan peregrines. 8(4): 255-260 Pecka, Z., Baulu, P., and Newsome, H. Preliminary survey of ethylenethiourea residues in the Canadian food supply, 1972. 8(4):232-234 Petrocelli, S. R., Anderson, J. W., and Hanks, A. R. DDT and dieldrin residues in selected biota from San Antonio Bay, Texas —1972. 8(3): 167-172 R Reidinger, R. F., Jr., and Crabtree, D. G. Organochlorine residues in golden eagles. United States— March 1964-July 1971. 8(l):37-43 Reimold, R. J,, and Durant, C. J. Toxaphene content of estuarine fauna and flora before, during, and after dredging toxaphene- contaminated sediments. 8(l):44-49 ROSENFELD, D., see Wassermann, M. Saha. J. G., see Fredeen, F. J. H. Sand, P. F., see Crockett, A. B. SCHNEEWEis, J. C, Greichus, Y. A., and Under, R. L. Organo- chlorine pesticide residue levels in North American timber wolves —1969-71. 8(2): 142-143 SCHULTZ, D. P., and Harman, P. D. Residues of 2,4-D in pond waters, mud, and fish, 1971. 8(3): 173-179 Sheets, T. J., see Jackson, M. D. SiLVA Fernandes, A. M. S., see Graca, I. Slach, E., see Mick, D. L. Smalley, H. E., see Clark, D. E. Tai, H., see Crockett, A. B. ToMATis, L., see Wassermann, M. w Ware, G. W., Estesen, B J., and Cahill, W. P. DDT moratorium in Arizona — agricultural residues after 4 years. 8(2):98-101 Wassermann, D., see Wassermann, M. Wassermann, M., Tomatis. L., Wassermann, D., Day, N. E., Groner, Y., Lazarovici, S., and Rosenfeld, D. Epidemiology of organochlorine insecticides in the adipose tissue of Israelis. 8(1): 1-7 White, C. M., see Peakall, D. B. WiERSMA, G. B., see Crockett, A. B. WiLDisH, D. J., see Zitko, V. Winterlin, W., Mourer, C, and Bailey, J. B. Degradation of four organophosphate insecticides in grape tissues. 8(l):59-65 Winterlin, W., Bailey, J. B., Lancbehn, L., and Mourer, C. Degra- dation of parathion applied to peach leaves. 8(4):263-269 WORLUND, D. D., see Anas, R. E. Yates, M. L., see Crump-Wiesner, H. J. Zitko, V., Choi, P. M. K.. Wildish, D. J , Monaghan, C. F., and Lister, N. A. Distribution of PCB and p,p'-DDE residues in Atlantic herring (Clupea harengus harengus) and yellow perch (Perca fiavescem) in eastern Canada. 8(2): 105-109 Vol. 8, No. 4. March 1975 279 Information for Contributors The Pesticides Monitoring Journal welcomes from all sources qualified data and interpretive information which contribute to the understanding and evaluation of pesticides and their residues in relation to man and his environment. The publication is distributed principally to scientists and technicians associated with pesticide monitoring, research, and other programs concerned with the fate of pesticides following their application. Additional circulation is maintained for persons with related in- terests, notably those in the agricultural, chemical manu- facturing, and food processing industries; medical and public health workers; and conservationists. Authors are responsible for the accuracy and validity of their data and interpretations, including tables, charts, and refer- ences. Accuracy, reliability, and limitations of the sam- pling and analytical methods employed must be clearly demonstrated through the use of appropriate procedures, such as recovery experiments at appropriate levels, confirmatory tests, internal standards, and inter-labora- tory checks. The procedure employed should be ref- erenced or outlined in brief form, and crucial points or modifications should be noted. Check or control samples should be employed where possible, and the sensitivity of the method should be given, particularly when very low levels of pesticides are being reported. Specific note should be made regarding correction of data for percent recoveries. Preparation of manuscripts should be in con- formance to the CBE Style Manual, 3d ed. Coun- cil of Biological Editors, Committee on Form and Style, American Institute of Biological Sciences, Washington, D. C, and/or the Style Manual of The United States Government Printing Office. An abstract (not to exceed 200 words) should accompany each manuscript submitted. All material should be submitted in duplicate (original and one carbon) and sent by first-class mail in flat form — not folded or rolled. Manuscripts should be typed on S'/i x 11 inch paper with generous margins on all sides, and each page should end with a completed paragraph. All copy, including tables and references, should be double spaced, and all pages should be num- bered. The first page of the manuscript must con- tain authors' full names listed under the title, with affiliations, and addresses footnoted below. Charts, illustrations, and tables, properly titled, should be appended at the end of the article with a notation in text to show where they should be inserted. li-U.S. Government Printing Office: 1975 — 621-567/3 -Charts should be drawn so the numbers and texts will be legible when considerably reduced for publication. All drawings should be done in black ink on plain white paper. -Photographs should be made on glossy paper. Details should be clear, but size is not important. -The "number system" should be used for litera- ture citations in the text. List references in the order in which they are cited in the text, giving name of author/ s/, year, full title of article, exact name of periodical, volume, and inclusive pages. The Journal also welcomes "brief" papers reporting monitoring data of a preliminary nature or studies of limited scope. A section entitled Briefs will be included, as necessary, to provide space for papers of this type to present timely and informative data. These papers must be limited in length to two journal pages (850 words) and should conform to the format for regular papers accepted by the Journal. Pesticides ordinarily should be identified by common or generic names approved by national scientific so- cieties. The first reference to a particular pesticide should be followed by the chemical or scientific name in parentheses — assigned in accordance with Chemical Abstracts nomenclature. Structural chemical formulas should be used when appropriate. Published data and information require prior approval by the Editorial Advisory Board; however, endorsement of published in- formation by any specific Federal agency is not intended or to be implied. Authors of accepted manuscripts will receive edited typescripts for approval before type is set. After publication, senior authors will be provided with 100 reprints. Manuscripts are received and reviewed with the under- standing that they previously have not been accepted for technical publication elsewhere. If a paper has been given or is intended for presentation at a meeting, or if a significant portion of its contents has been published or submitted for publication elsewhere, notations of such should be provided. Correspondence on editorial matters or circulation mat- ters relating to official subscriptions should be addressed to: Paul Fuschini, Editorial Manager, PESTICIDES MONITORING JOURNAL, Technical Services Divi- sion, Office of Pesticides Programs, U. S. Environmental Protection Agency, Room B49 East, Waterside Mall, 401 M Street, S.W., Washington, D. C. 20460. 280 Pesticides Monitoring Journal BOSTON PUBLIC LIBRARY 3 9999 05571 195 4