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Foreword

The first issue of the "Pesticides Monitoring
Joiirnal" is a direct result of cooperation among
four Federal Departments, each of which is
responsible for a distinctive mission in the field
of pesticide usage. Because the ultimate objec-
tive of all of these programs is the enhancement
of man's welfare, their collaboration is in the
best public interest.

The initiative for this joint effort came from
the Departments themselves. In 1961, the Sec-
retaries of Defense, Interior, Agriculture, and
Health, Education, and Welfare undertook the
formation of the Federal Pest Control Revieiv
Board with the intent that it would review ". . .
the various programs conducted by Federal
agencies for control of forms of invertebrate
and plant life which adversely affect man's in-
terests, and 'shall' consider problems and devel-
opments in the field of chemical control, loith
particular reference to possible adverse effects
and the adequacy of provisions for the proper
use of pesticidal chemicals to insure the greatest
public and national benefit." The Board ivas
directed to turn its attention to all aspects of
pest control, including the need; safety to man,
domestic animals, ivildlife, and the environment
in general; and alternative methods. The Board
ivas instructed to advise the Departments on
modifications in plans that ivould be in the best
public interest in view of these and related
matters.

In 196 Jf, in response to the report of the Presi-
dent's Science Advisory Committee on "Use of

Pesticides" and with the advice and encourage-
ment of the Executive Office of the President,
especially the Office of Science and Technology
and the Bureau of the Budget, these four Sec-
retaries reorganized the Board as the Federal
Committee on Pest Control. The reorganization
was necessary to expand the collaboration in
two directions: first, to permit the new Com-
mittee to cover all aspects of pest control —
research, monitoring of the environment for
pesticides, and public information programs —
as ivell as to review operational programs; sec-
ondly, to extend their council to all Federal pro-
grams involving pests and their control.

The "Pesticides Monitoring Journal" is an out-
growth of one of the recommendations of the
President's Science Advisory Committee that
the concerned agencies "develop a continuing
network to monitor residue levels in air, water,
soil, man, wildlife, and fish."

To implement this recommendation the Federal
Committee on Pest Control established a Sub-
committee on Pesticide Monitoring which peri-
odically evaluates the activities in this area
throughout the Nation. Much of the work of
monitoring levels of pesticides in the environ-
ment is being done by universities, State Agri-
cultural Experiment Stations, conservation
groups, and other non-Federal agencies. The re-
sults of many of these studies are not published,
or appear in journals or individual reports that
are scattered and difficult to locate. For this
reason, the Subcommittee recommended that a
Journal be established to assure accessibility of
monitoring data to the scientists who need it.
It is hoped that such data will not only provide
information on the present levels of pesticide
residues in various elements of the environment,
including man, but will provide a base line from
which we can determine whether these levels
are increasing, decreasing, or remaining sub-
stantially unchanged.

There is an inherent risk in such an endeavor.
Data of the type which tvill appear in this
Journal are subject to misinterpretation. The
significance of "parts per million" levels of a
given chemical in the soil or in river water is
not entirely clear. There is disagreement as to
^vhether or not a particular level in a particular
place at a particular time represents a signifi-

A

^^ lH.^-.(-3

cant hazard to man or u-ildlife, or other envi-
ronmental components. If then, such tifjitrcs are
published in a journal, is there not a possibility
that special interest groups will quote them
to "prove" their oirn preconceived biases?
The Federal Covimittee on Pest Control has
decided that such a risk must be taken. The
alternative would be to encourage the scientists
who gather such data to release only their oivn
interpretations; hoivever, no matter how ivell
intentioned, such interpretations would not
necessarily provide a sound basis for evaluation
of changes in levels that may occur in the future.
It will be the intent of this Journal to publish
the data in a form that will permit each reader
to interpret the results for himself. Information
on sampling procedures a7id analytical methods
used to gather each set of data will be included.
Through this interdepartmental venture, the
Federal Committee on Pest Control is demon-
strating the practicability of collaboration be-
ttveen agencies with such diverse missions as
food production, disease prevention, protection
of human health and food supplies, and conser-
vation of our nxitural resources.

The Committee has no direct appropriation to
undertake such a venture. Therefore, the
responsibility for staffing and financing this
Journal has been delegated to one of the mem-
ber agencies, but the editorial policy and guid-
ance will continue to be the responsibility of an
Editorial Board ivith members drawn from six
different agencies of three of the cooperating
Departments. The Editorial Board is appointed
by and responsible to the Federal Committee on
Pest Control. Thtis the initiative of the four
Federal Departments in establishing this Com-
mittee has been snatched by the ingenuity of
the Committee itself in finding a method of im-
plementing programs without proliferating new
authorities and agencies. It is for these reasons
that the Office of Science and Technology has
encouraged the Federal Committee on Pest
Control to look beyond the four initiating
Departments and to advise on all aspects of
pests and their control in which any agency of
the Federal Government is involved.

Ivan L. Bennett, Jr., M. D.

iJoputy I>i rector.

Office of Science and TechnoloKy,

Executive Office of the President

CONTENTS

Volume 1

June 1967 Number 1

Foreword

Ivan L. Bennett, Jr.

Introduction

John M. Geary

NATIONAL PESTICIDE
MONITORING PROGRAM

Page
Residues in Food and Feed

Assessments include raw food and feed com-
modities, market basket items prepared for

consumption, meat samples taken at slaughter „.. 1

R. E. Duggan and F. J. McFarland

Pesticides in People

Criteria for monitoring pesticides in people in-
clude high- and low-exposure conditions, age,

sex differences — 6

Anne R. Yobs

Residues in Fish, Wildlife, and Estuaries

Indicator species near top of food chain chosen
for assessment of pesticide base levels in fish
and wildlife — clams, oysters, and sediment
in estuarine environment „. 7

R. E. Johnson, T. C. Car\'er, and E. H.

Dustman

Pesticides in Water

Network to monitor hydrologic environment

covers major drainage rivers 13

R. S. Green and S. K. Love

Pesticides in Soil

National soil monitoring program studies high-,

low-, and nonuse areas _ -16

P. F. Sand, J. W. Gentrj', J. Bongberg, and

M. S. Schechter

Chemicals Monitoring Guide for the National

Pesticide Monitoring Program -20

Milton S. Schechter

The Pesticides Monitoring Journal is published quarterly under the auspices of the

Federal Committee on Pest Control and its Subcommittee on Pesticide Monitoring as a

source of information on pesticide levels relative to man and his environment.

The parent committee is composed of representatives of the U. S. Department of

Agriculture, Defense, the Interior, and Health, Education and Welfare.

The Pesticide Monitoring Subcommittee consists of representatives of the Agricultural

Research Service, Consumer and Marketing Service, Federal Extension Service,

Forest Service, Department of Defense, Fish and Wildlife Service, Geological Survey,

Federal Water Pollution Control Administration, Food and Drug Administration,

Public Health Service, and the Tennessee Valley Authority.

Responsibility for publishing the Pesticides Monitoring Journal has been accepted

by the Pesticides Program of the Public Health Service.

Pesticide monitoring activities of the Federal Government, particularly in those agencies

represented on the Pesticide Monitoring Subcommittee which participate in operation

of the national pesticides monitoring network, are e.xpected to be 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 non-government research institutions, both within and without the United

States. 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 Subcommittee. Authors are given the benefit of review

comments prior to publication.

Editorial Advisory Board members are:

Reo E. Duggan, Food and Drug Administration, Chairman

Anne R. Yobs, Public Health Service

James B. DeWitt, Fish and Wildlife Service

Richard S. Green, Federal Water Pollution Control Administration

S. Kenneth Love, Geological Survey

Milton S. Schechter, Agricultttral Research Service

Paul F. Sand, Agricultural Research Service

Trade names appearing in the Pesticides Monitoring Journal are for
identification only and do not represent endorsement by any Federal
agency.

Address correspondence to:

Editorial Manager
PESTICIDES MONITORING JOURNAL

Pesticides Program

National Communicable Disease Center

Atlanta, Georgia 30333

-J

INTRODUCTION

PESTICIDES

AND

THE TOTAL

ENVIRONMENI

John M. Geary'

The Federal Committee on Pest Control is con-
cerned with assuring necessary control of pests
without hazard to the environment and its
inhabitants, including man. The Committee,
while encouraging the use of other types of
control, recognizes that chemical methods will
continue to be needed for some time to come. It
is therefore necessary to evaluate the long-term
effects of such chemicals and their residues in
the environment.

' Chairmnn. Kc<lcrnl Committee on Pest Control. Walter Keed Army
.Me<llral Center. Waihinirlon, D. C. 20012.

The effects of pesticides may be directly on
organisms that are the target of control or on
closely associated organisms. The effects may
also be indirect or considerably delayed, with
a certain amount of movement of pesticides in
the environment after application. To evaluate
the indirect effects it is necessary to know
something about the distribution of pesticides
in the various elements of the environment and
the changes in these levels with time. The
determination of this information is what the
Federal Committee on Pest Control considers
to be "pesticide monitoring."

The application of pesticides, depending on the
target of control, may contaminate air, water,
soil, plants, wildlife, and man. That portion
which gets into the air may then settle on other
parts of the environment or be carried for a
considerable distance in air currents. It is likely
that only a small amount will become uniformly
distributed throughout a large mass of air. The
selection of truly representative air samples is
difficult even if the concentrations are large
enough to make chemical detection easy.

The distribution of contaminating pesticides
that may settle on water depends upon many
factoi's — the solubility or suspendibility of the
formulation in water, the movement of the
water as well as its physical and chemical
characteristics, and the presence of biological
organisms.

Pesticides settling on soil may remain on the
surface and later be moved by wind or washed
off by rain — or they may penetrate to some
depth. Penetration will depend on the character-
istics of the material and the soil as well as on
rainfall and other conditions. A portion of the
pesticides in soil may be absorbed by plants or
other organisms, and a small amount may be
translocated.

Pesticides falling on plant surfaces may directly
affect the plant, degrade with weathering, wash
off into soil or water, or remain as residues.
Residues may be carried away with falling
leaves, consumed by animals, or harvested with
crops. Ill the latter case they may be redistrib-
uted dui'ing processing or be ingested by the
final consumer.

Pesticides making primary contact with man
and animals may be absorbed and stored or

excreted — or they may eventually reach other
parts of the environment. Even that portion
that reaches man directly, through handling of
pesticides or by drift, may be washed off the
skin and contaminate the soil or water.

An important consideration in the complex
problem of the physical distribution of pesti-
cides is the length of time required for break-
down of pesticides to other compounds. The
time required for such degradation varies with
each particular material. Some pesticides hydro-
lyze in the presence of moisture, and others may
oxidize in air. Sunlight may act as a catalyst in
decomposition, and this effect will certainly be
modified by the concentration of the pesticide as
well as by the amount and nature of the light.
If pesticides occur in large aggregates, are ab-
sorbed in solid particles, or are deeply buried
in the ground, the decomposition may be radi-
cally affected.

Finally, pesticides may be decomposed by bio-
logical processes. Such decomposition may vary
greatly between organisms and may also be
affected by chemical and physical conditions.

The interpretation of monitoring data must
always be restricted to the exact portion of the
environment of which it is representative.
Within this limitation, the reliability of the
interpretation will be affected by the adequacy
of the sampling design, and the sensitivity and
accuracy of the analytical procedures employed.

The many complexities of monitoring make it
clear that there can be no simple and prompt
answers to the questions of what pesticide
residues are now present in our environment,
where they are, and at what levels. The Federal
Committee on Pest Control feels that such data
must be permanently recorded in a form that
will permit comparison between different
studies and must be readily available to those
who need such information. It is for these rea-
sons that the "Pesticides Monitoring Journal"
was proposed.

It is the intention of the Committee and the
Editorial Board that the data published in the
Journal be sufficiently detailed to show the
precise portion of the environment sampled and
to enable the user to judge the accuracy and
dependability of the work. The reliability of the
sampling, handling, cleanup, and chemical anal-

yses is, of course, the responsibility of the
author of each paper. The Editorial Board can
only reject those papers in which the presenta-
tion leaves some doubt as to the adequacy of
these procedures. The competence of the reader
will be relied upon to avoid unwarranted gen-
eralizations from data that represent only a
small segment of the total environment.

In an effort to provide a minimal base from
which adequate information can be gained, the
Subcommittee on Pesticide Monitoring recom-
mended a national pesticide monitoring pro-
gram which is described in this first issue of
the "Pesticides Monitoring Journal." In addi-
tion to reporting the results of the national
program, the Journal should serve as the pub-
lishing medium for essentially all pesticide
monitoring efforts in this country. This should
include individual work, as well as large pro-
grams, and foreign contributions also will be
welcomed. The degree to which the goals of the
Journal can be achieved will depend on the coop-
eration of interested workers.

The Federal Committee on Pest Control is the
sponsor of the "Pesticides Monitoring Journal,"
but it was the members of the Subcommittee on
Pesticide Monitoring who contributed long
hours of devoted work to its establishment and
to the initiation of the national program. The
Subcommittee is made up of the people who
most adequately represent the philosophy of
the Federal government as to what a national
pi-ogram and journal should be.
The Federal Committee on Pest Control is not
an independent agency of the Federal govern-
ment with its own operating funds, but is an
association of personnel designated by the Sec-
retaries of the four departments which are
most concerned with pesticides. Therefoi'e, the
actual publication of the Journal cannot be
undertaken by the Committee. The Department
of Health, Education, and Welfare, one of the
member departments of the Committee, has
agreed to be the publisher, through the Pesti-
cides Program of the U. S. Pubhc Health Service,
National Communicable Disease Center, Atlan-
ta, Georgia. The Federal Committee on Pest
Control appreciates this cooperation and ex-
tends its thanks. The Federal Committee on
Pest Control remains responsible for editorial
policies, guidance, and general content.

IHTIONAl
FESrieiDE

Monitoring

PROERAM

This initial issue of the PESTICIDES MONITORING
JOURNAL is devoted in its entirety to a description
of the National Pesticide Monitoring Program as rec-
ommended by the Subcommittee on Pesticide Monitor-
ing of the Federal Committee on Pest Control and es-
tablished by the responsible agencies. For the most
part, the program as described represents the mini-
mum effort necessary for adequate assessment of
pesticide levels in man and his environment- it does
not include all of the monitoring activities being
conducted by the various Federal agencies.

Publication of original data derived from the National
Pesticide Monitoring Program and from other Federal
and non-Federal monitoring programs will commence
with the second issue.

RESIDUES IN
FOOD AND FEED

ASSESSMENTS INCLUDE RAW
FOOD AND FEED COMMODITIES,
MARKET BASKET ITEMS
PREPARED FOR CONSUMPTION,
MEAT SAMPLES TAKEN AT
SLAUGHTER

R. E. Duggani and F. J. McFarland-

The Federal program for monitoring pesticide
residues in food and feed primarily is comprised
of surveillance programs maintained by the
Food and Drug Administration, U. S. Depart-
ment of Health, Education, and Welfare. Data
on residues in meat samples will be provided by
the Livestock Slatighter Inspection Division,
Consumer and Marketing Service, U. S. Depart-
ment of Agriculture.

Monitoring Objective

The objective of this program is to determine
the levels of pesticide residues in unprocessed
and commercially processed consumer food com-
modities, animal feeds, and composites of food
items prepared for human consumption. Studies
being carried out to accomplisli this objective
include (1) a continuing Market Basket study
to assay pesticide residues in the basic 2-week
diet of a 19-year-old male, statistically the
Nation's largest eater, and (2) nationwide sur-
veillance of unprocessed food and feed. In addi-
tion, the Livestock Slaughter Inspection Divi-
sion of the Consumer and Marketing Service,
U. S. Department of Agriculture, will provide
significant data on the analysis of meat samples
taken from animals at slaughter.

Factors Influencing Program Design

Numerous interrelated factors necessarily have
been considered and evaluated in defining a
minimum monitoring efi'ort for pesticide resi-
dues in food and feed.

Many individual commodities entering the
Nation's food supply are produced in various

1 Office of Associate Commissioner for Compliance, Food and DruK
Administration. U. S. Department of Health, Education, and Wel-
fare, Washinirton, D. C. 20204.

- Bureau of Science, Food and Drucr Administration, U. S. Depart-
ment of Health. Education, and Welfare. Washington. D. C. 20204.

geographical areas. Because the distribution
system which brings these commodities to mar-
ket is rapid, a constant shifting of commodity
origins exists within a given consumption area.
Since there are no crossroads in time or geo-
graphy to permit concentrated or liighly selec-
tive sampling which could be considered
sufficiently representative of the food supply,
monitoring of residues in food and feed must be
programmed on a continuing and broadly
geographical basis.

It should be recognized that the important im-
pact of pesticide residues in human and animal
food, insofar as environmental effects are con-
cerned, lies in their consumption. Therefore,
the examination of foods as they are prepared
and ready for consumption is of special interest
to this monitoring program. Residues in wastes
from food processing, of course, may be of con-
cern with regard to soil, water, or the atmos-
phere, depending upon their final disposition.
Their efl'ect on these elements of the environ-
ment, however, would be detected by other
monitoring programs.

Because no unifoiTnity may be expected within
even a single food item due to extreme varia-
tions in local growing, harvesting, and proces-
sing procedures, sampling patterns taking
geographical and seasonal variables into account
must be used. Moreover, examination of the 82
individual food items in the Market Basket
Survey was considered impractical because of
the spectrum of unknown residues potentially
present and the limitations in analytical meth-
ods to detect and measure moi'e than one class
of residues. The dilution factor, technical prob-
lems in methodology, and variations in dietary
habits suggested that composites representing
a "total diet" also would be unsatisfactory. To
minimize these problems, a practical compro-
mise was reached, that is, the compositing of
foods by classes, e.g., meats, dairy products,
green vegetables, etc.

Data yielded by this method, especially when
correlated with that obtained on unprocessed
foods, may be used to calculate the approximate
residue intake associated with any diet pattern.
Such correlations, however, would be a subject
for special research projects and are not specifi-
cally contemplated as a function of the monitor-
ing program.

Vol. I, No. 1, June 1967

Geographical Disti'ibution of
Sampling Stations

Sampling in the Market Basket Survey (for
analysis of composites of food items prepai-ed
for consumption) is carried out in five regions
representing the northeastern, soutlieastern,
north central, central, and western United
States. Sampling sites within each region are
chosen from different cities, one representing a
stiindard metropolitan statistical area and one
representing a smaller population center (less
that 50,000 population).

Samples in the nationwide surveillance of un-
processed foods are collected at all major grow-
ing, processing, and marketing centers. Animal
food ready for consumption is included in this
part of the progi'am. Collection headquarters
are in each of the 18 Regional Districts of the
Food and Drug Administration, with offices in
Boston, New York City, Bufi'alo, Philadelphia,
Baltimore, Atlanta, Cincinnati, Detroit, Chi-
cago, St. Louis, New Orleans, Minneapolis,
Kansas City, Dallas, Denver, Los Angeles, San
Francisco, and Seattle.

Meat samples at slaughter will be taken at each
of the Nation's major meat processing centers.

Sampling Frequency, Number of Samples

Market Basket samples are collected six times
per year in each of the five geographic regions
mentioned above, making a total of 30 Market
Basket samples annually.

The surveillance program encompasses an esti-
mated 2.5 million carloads of raw agricultural
products annually shipped in interstate com-
merce. In addition, there are thousands of lots
of other foods, e.g., milk, eggs, fish, and pro-
cessed animal feeds, produced each year.

A minimum coverage sampling procedure has
been designed whereby product samples will be
collected throughout the year in the following
broad categories :

Leaf and stem vegetables Fish and shellfish

Root vegetables Eggs and egg products

Fruits Fluid Milk

Grains Manufactured Dairy
flay and silage Products

Sampling locations are selected at random from
wholesale markets and warehouses located in
85 cities. Approximately 12,000 random sam-

ples^ are examined annually. This provides 95%
confidence that the true percentage of samples
exceeding guidelines will not be greater than
3.1% if the observed percentage is 2%. When
the observed percentage of samples exceeding
guides approaches 3''r , the sampling rate is in-
creased to provide more reliable estimates.

Commodities to be Sampled

In the Market Basket Survey, samples are
collected according to a series of 82 items listed
by commodity groups in Appendix A. Adjust-
ments are made in this list to reflect local
dietary patterns in each geographical area. The
list also will be evaluated periodically and
changed as necessary to reflect changes in
dietary patterns, particularly in the area of
"convenience" and frozen foods.

Commodities sampled under the nationwide
surveillance of unprocessed foods are listed in
Appendix C. A sampling schedule for these
commodities is included as Appendix D.

Sample Preparation

Market Basket items which normally require
further processing by cooking, such as fresh
meats and certain raw vegetables, or prepara-
tion for eating raw, such as tomatoes, carrots,
celery, lettuce, cucumber, cabbage, and fresh
fruits are delivered to a diet kitchen for
preparation under the direction of a dietician.
Some food items, e.g., cabbage, are included in
both the raw and cooked foiTns. Instructions to
the diet kitchen for preparing these food items
are contained in Appendix B.

Market Basket items normally consumed as pur-
chased or which do not otherwise require
further processing — e.g., dairy products, lunch-
eon meats and frankfurters, canned meats,
some fruits and vegetables, potato chips, canned
fruit juices, concentrated fruit juices, and fro-
zen fruits — are to he retained by the examining
laboratory for compositing by commodity
groups with the foods prepai'ed by the diet
kitchen.

Guidelines for compositing food items sampled

■' The IViod .in<l Druj^ A<!ministration also examines about the same
number of samples selected liocause of susi>ecte<i residues as an
accompanyinjT ]»art of its enforcement activities. This is not con-
sidered a monitoring activity, and the prot:rani is not described.

Pesticides Monitoring Journal

in the surveillance of unprocessed foods are
given in Appendix E.

Sample Atmlysis Procedures

All analytical procedures used in this program

are described in FDA's Pesticide Analytical

Manuals.

For the Market Basket Survey, procedures for
examining each commodity group are outlined
as follows:

1. Chlorinated Organic Pesticides ■ — Examine all
commodity groups at sensitivity levels equivalent
to 0.003 parts per million heptacldor epoxide using
electron capture, gas-liquid chromatography. Resi-
dues above these limits are to be checked by thin-
layer chromatography or Dohmnann glc, and re-
stdts reported to the nearest 0.001 ppm. Multiple
detection procedures are to be used to detect the
25 chlorinated organic compounds included in Ap-
pendix F.

2. Organic Phosphate Pesticides — Examine all
commodity groups in conjunction with chlorinated
organic residue analyses at a sensitivity level of
0.05 ppm of parathion. Confirm positive findings
by thin-layer chromatography.
Herbicides — Examine all commodity groups by
Dohrmann glc, confirvi by paper chromatography.
Examine all commodity groups in Appendix A,
except groups 1, 2, and 10 for .S-AT (3-aminotri-
azole). Confirm residues by paper chromatography.
Carbamates — Examine all cotnmodity groups for
carbaryl, except groups 1, 2, and 10, Appendix A,
and confirm positive findings. Use general methods
for dithiocarbamates in examining above groups
and report residts as zineb.
Bromides — Examine all commodity groups.
Arsenic — Examine all commodity groups.

For the nationwide surveillance of unprocessed
food and feed, all samples are to be examined
for chlorinated organic pesticides and organic
phosphate pesticides (See Appendix F) using
multiple detection procedures at sensitivity
levels equivalent to 0.03 ppm heptachlor epoxide
using electron capture, gas-liquid chromatogra-
phy. Individual samples selected at random are
to be examined for residues of chlorophenoxy
compounds, carbaryl, and carbamates. Analyt-
ical procedures are described in the Food and
Drug Administration's Pesticide Analytical
Manuals.

s.

u.

5.
6.

Appendix A

MARKET BASKET COMPOSITION BY COMMODITY GROUPS

1. Dairy Products

Cottage Cheese

Processed Cheese (American)

Milk, Fresh Fluid

Evaporated Milk

Natural Cheese

Nonfat Dry Milk

Butter or (alternate)

Ice Cream

Margarine

i. Meat, Fish, anil Poultry

Green Beans, raw
Green Beans, frozen
Green Beans, canned
Beans, w/pork, canned
Lima Beans, raw
Lima Beans, canned
Lima Beans, frozen

Lamb or Mutton

Roast Beef

Ground Beef

Pork Chops

Pork Sausage (cured)

Bacon

Chicken (eviscerated — fresh

or frozen)
Fish Fillet, fresh or frozen
Tuna or Salmon, canned
Luncheon Meat
Frankfurters
Liver, beef
Eggs, large

7. Root Vegetables

Carrots, raw
Carrots, canned
Onions, dry, (raw V2)
Onions, dry, (boil ',2)
Beets, w/o tops, raw
Beets, w/o tops, canned
Turnips, w/o tops, raw
Rutabagas, raw

3. Grain and Cereal Products

Flour, General Purpose
Flour, Self-Rising
Pancake Mix
Corn Flakes
Shredded Wheat

or Wheat Cereal
Rice Flakes or Puffed Rice
Oatmeal
Rice

Corn Meal
Macaroni, elbow
Bread, White (enriched)
Bread, White (unenriched)
Bread, Italian style
Bread, Whole Wheat
Rolls (sweei, cinnamon,

Bismarcks, doughnuts)
Saltines
Cookies, Plain
Rolls (frankfurter)
Graham Crackers
Pie Crust

(fruit filling — item 9)
Cake
Corn, raw
Corn, canned
Corn, frozen

8. Garden Fruits

Pepper, raw
Tomatoes, fresh
Tomatoes, canned
Catsup

Cucumbers, raw
Eggplant, raw
Summer Squash, raw
Summer Squash, frozen

9. Fruits

Fruit filling from pie
Oranges, fresh
Orange Juice

frozen cone,

canned
Bananas
Raisins
Peaches, fresh
Peaches, canned
Apples, fresh
Apples, canned
Strawberries, raw-fresh

or frozen
Apricots, raw
Apricots, canned
Cherries, raw
Cherries, canned
Cherries, frozen
Grapes, raw
Pears, raw
Pears, canned
Pineapple, raw
Pineapple, canned
Pineapple, frozen
Plums, raw
Plums, canned
Rhubarb, w/o top,

raw-fresh or frozen
Watermelon, raw

4. Potatoes

Potatoes, white (bake V,)
Potatoes, white (boil \i)
Potatoes, white (fry Vt)
Potato Chips
Sweet Potatoes or Yams,
fresh or canned

i. Leafy Vegetables

Beet Tops, raw
Beet Tops, canned
Collards, raw
Collards, canned
Mustard, raw
Mustard, canned
Spinach, raw
Spinach, canned
Spinach, frozen
Celery, raw
Lettuce, raw
Cabbage (raw V2)
Cabbage (boil V2)
Broccoli, fresh
Broccoli, frozen
Asparagus, canned
Asparagus, fresh
Asparagus, frozen
Mushrooms, raw
Mushrooms, canned
Cauliflower, raw
Cauliflower, frozen

10. Oils, Fats, and Shortening

Salad Dressing - French

Mayonnaise

Salad Oil

Shortening

Peanut Butter

11. Sugar and Adjuncts

Sugar, White
Jam or Jelly
Pudding Mix
Syrup, blended
Molasses
Candy Bar
Baking Powder
Salt
Vinegar, cider

12. Beverages

Tea Leaves
Cocoa

Drinking Water
Coffee, ground
Soft Drinks

S. Legume Vegetables

Peas, raw
Peas, canned
Peas, frozen

Vol. I, No. 1, June 1967

Appendix B

INSTRUCTIONS FOR FOOD PREPARATION AND CHECK LIST OF ITEMS IN SAMPLE (Market BasKet Surrey)
The food items listut Inlow an those requiring preparation. The preparation may co7isist of roasting, baking, broil-
ing, frying, or boiling. Some vegetables are to be prepared to eat raw. After processing, wrap in aluminum foil and
place in labeled containers.

FOOD ITEM
Roast heel

Ground Bee/
Pork chops

Pork sausage

Bacon

Chicken

Fish fillet
Liver, beet
Potatoes, white

Tomatoes. Iresh
Oranges, raw
Carrots, raw
Greens

(Beet tops,

Collards, Mustard,

Spinach)
Green Pepper
or broccoli

Sweet potatoes

Celery

Lettuce

Cucumber

Cabbage

Onions, dry

Peas

INSTRUCTIONS

Roast, medium-well rfo?ie. Remove bone
and discard. Save drippings.
Make into patties, broil, save drippings.
Broil. Remove bone and discard. Save
drippings.

Make into patties, broil.
Broil.

Discard neck and tail portion. Bake. Re-
move edible meat from bone a/ter baking.
Save drippings.
Broil.

Broil, save drippings.
Bake. Leave skin on. (5 lbs.)
Fry. (2'.'2 lbs.)

Boil. Peel and discard skin before boihng.
(2^2 lbs.) Discard cooking water.
Wash, remove core, do not peel.
Remove peel and seeds.
Wash, scrape, slice ready-to-eat raw.
Fresh or frozen. Wash, trim, cook fresh
item. Cook frozen item. Discard cooking
water.

One item only. Pepper, fresh. Broccoli,

fresh or frozen. Pepper, prepare to eat

raw. Fresh broccoli — wash, trim, and cook.

Frozen broccoli, cook. Discard cooking

water.

Wash, bake, and peel.

Wash, trim, cut.

Trim, quarter.

Wash with detergent to remove wax.

(1) Raw. Trim and chop for slaw.

(2) Cook after trimming. Discard cooking
water.

(1) Raw. Clean and quarter.

(2) Cooked. Clean and boil. Discard
cooking water.

Fresli i7i season. Remove pods, cook.
Frozen, cook. Discard cooking water from
both.

Green Beans
Com, sioeet

Peaches, raw

Apples
Strawberries

Other Vegetables
Asparagus

Beets

Mushrooms

Turnips
Lima Bearis

Cauliflower

Eggplant
Rutabagas

Summer Squash

Other Fruits

Apricots

Cherries

Crapes

Pears

Pineapple

Plums

Rhubarb

Watermelon

Cooking Oil

Fresh if available. Wa-th and cook fresh
or frozen. Discard cooking leater.
Fresh if available. Remove husk, trim,
cook iji boiling water. Discard water. Re-
move cooked corn from ear. Cook frozen
corn and discard water. Discard cobs.
Fresh when available. Wash, peel, remove
pits, and halve.

Wash, remove core, do not peel.
Fresh in season. Wash, remove stems.
Halve.

Fresh and frozen vegetables will be cooked.

Fresli in season. Wasli and cook. Frozen,
cook. Discard cooking water.
Fresh beets. Wash, trim, and cook. Dis-
card cooking water.

Wa.ih. trim, and boil. Discard cooking
'water.

Wash, trim, and cook. Discard cooking
water.

Fresh, shell and cook. Frozen, cook. Dis-
card cooking water.

Fresh, wash, trim, and cook. Frozen, cook.
Discard cooking water.
Trim and cook. Discard cooking water.
Trim and cook. Discard cooking water.
Fresh, trim and cook. Frozen, cook. Dis-
card cooking water.

Fresh fruits only to be processed.

Wash and pit.
Wasli and pit.

Wash, remove seeds, and stems.
Wash and core.
Trim and core.
Wash and pit.
Trim.

Trim and remove seeds.
(Unused cooking oil to be returned with
processed foods and included in com-
posite.)

Appendix C

NATIONWIDE SURVEILLANCE

COMMODITIES

Large fruit

Small fruit

Leaf and stem vegetables

Vine and ear vegetables

Beans

Root vegetables

Nuts

Hay and silage

Wheat

Corn

Oats

Rye

Sorghum

Barley

Flax

Rice

Soybeans

Fluid Milk

Shell Eggs

Fish and Oysters

Meats (beef, pork, mutton,
lamb, and poultry)

Mfgd. Animal Feed

Vegetable Oil

Fish Liver Oil

Other [e.g., coffee beans,
cocoa beans, spices (black
pepper, ginger) paprika, etc.)

Appendix D

SAMPLING SCHEDULE FOR NATIONWIDE
SURVEILLANCE COMMODITIES

Treat each identifiable grower's mark
or lot number in the shipment as a
separate sample. Sample, as a single
lot, shipments containing commingled
and uyiidcntifiable lots from several
growers. Be careful not to collect more
than the proportional amount from
facing layers. When sampling from
loading cars, select subsamplcs at in-
tervals to obtain a sample represen-
tative of the carload. For bulk lots
select subs at random throuqhout the
lot.

Collect a composite .sample closely ap-
proximating 20 lbs. by taking a S lb.
sub from each of 10 different shipping

containers selected at random. DO
NOT cut or divide individual produce
items to adjust sub weights.
SPECIAL NOTE: Some produce
items wcigliing 2 lbs. or more each,
such as melons, pineapples, large
heads of cabbage, large cauliflower,
large celery stalks, large rutabagas,
etc., do not lend themselves to the
above sampling approaches. In such
cases, collect a total composite sample
of 10 subs taking one item from each
shipping container.

For light bulky produce, e.g., collards,
spinach, leaf lettuce, other leafy prod-
ucts, hay, etc., collect a 10-lb. com-
posite sample taking 1 lb. from each
of 10 different shipping containers
selected at randonn.
Hold samples in cold .storage until
ready to be shipped or delivered to the
laboratory only if normally held or
shipped under refrigeration in com-
mercial practice.

Pesticides Monitoring Journal

Appendix E

GUIDELINES FOR COMPOSITING UNPROCESSED FOOD SAMPLES

ANIMAL TISSUE

DAIRY PRODUCTS

EGGS

FEED, ANIMAL

FORAGE

FRUITS

GRAINS

HAY
MILK

NUTS

OILS
SEEDS
SPICES
VEGETABLES

LARGE

SMALL

HEAD

LEAFY

POD

ROOT

STALK

Appendix F

Grind about nalj of each sub (-meat
grinder); composite 100 g from, each sub
and grind again.

Equal weight from, each sub. Grind, dice,
or blend.

Equal number of units from each sub,
for total of 6-12. Blend.

200 g from each sub (quarter subs down
to 200 g where necesary); wet feeds (sil-
age) 100 g from each sub.

Quarter each sub down to 200 g: compos-
ite 200 g from each sub. Chop fine. Where
necessary, grind in Wiley Mill without
screen; then with screen in.

(apples, pears, tomatoes, etc.). Equal

number of units from each sub. Chop or

blend.

200 g from each sub. Chop or blend.

100 g from each sub. Grind in Wiley Mill

or equivalent.

200 g from each sub. Chop or grind.

100 g (ml) from each sub after thorough

shaking.

Remove shells. Composite equal number
of units (equal weight) from each sub.

Chop or grind.

Equal weight or volume from each sub.
100 g from each sub. Grind.
200 g from each sub. Grind or chop.
Quarter each head in the sub. Take two
opposite quarters from each head and
chop into I- to 2-inch pieces with a knife.
Mix well. Composite 200 g of chopped
product from each sub and chop entire
composite in a food chopper.
Leaf Cut — Mix sub well and select leaves
at random until a 200-g portion is ob-
tained. Composite in a food chopper,
(beans, peas, etc., also asparagus) 200 g
from each sub. Chop or grind.
Equal number of units from each sub.
Chop or grind.

(celery, broccoli, etc.) Quarter each sub
lengthwise and proceed as in "Head Veg-
etables."

aUANTITATlVE AND QUALITATIVE
COMMONOR TRADENAME

CHEMICAL NAME

1. Aldrin

BHC (benzene
hexachloride)

3. Bulan®

4. Butyl ether ester,
2,4-D

5. n-Butyl ester, 2,4-D

6. n-Butyl ester,
2,4,5-T

7. Chlorbenside

8. Chlorobenzilate

1 2 3,4,10,10-hexachloro-l,4,4a,5.8,8a-

hexahydro-l,4-endo-eio-5,8-dimethano=

naphthalene

1 ,2 .3 ,4 ,5,6-hexachlorocyclohexane
2-nltro-l,l-bis(p-chlorophenyl) butane

butyl ether ester of 2,4-dlchlorophenoxy=

acetic acid

7j-butyl ester ot 2,4-dichlorophenoxyacetic

acid

n-butyl ester of 2,4,5-trlchlorophenoxyacetlc

acid

p-chlorobenzyl-p-chlorophenyl sulfide

ethyl 4,4'-dichlorobenzllate

9. Chlordane

10. Chlorothion

11. CIPC

12. Dacthal®

13. DDE

14. DDT lo,p'+p,p';
o,p'; p.p')

15. Diazinon

16. Dichloran

17. Dieldrin

18. Dilan (See Bulan®
and Prolan®)

19. Dyrene®

20. Endrin

21. Ethion

22. Ethyl hexyl ester,
2,4-D

24. Folpet

25. Heptachlor

26. Heptachlor Epoxide

27. Hexachlorobenzene

28. Isobutyl ester,
2,4-D

29. Iso-octyl ester
2,4,5-T

30. Iso-octyl ester,
2,4-D

31. Isopropyl ester,
2,4,5-T

32. Isopropyl ester,
2,4-D

33. Kelthane®

34. Lindane

35. Malathion

36. Methoxychlor

37. Methyl parathion

38. Ovex

39. Parathion

40. PCNB

41. Perthane® & olefin

42. Prolan®

43. Ronnel

44. Strobane®

45. TCNB

46. TDE

47. Tedion®

48. Telodrin®

49. Tetraiodoethylene

50. Thimet®

51. Thiodan I®

52. Toxaphene

53. Trithion®

54. Vegadex®

l,2,3,5,6.7,8,B-octachloro-2.3.3a,4,7,7a-
hexahydro-4,7-methanolndene

0.0-dlmethyl 0(3-chloro-4-nitrophenyl) =

phosphorothloate

Isopropyl N- (3-chlorophenyl) carbamate

dimethyl 2,3,5,6-tetrachloroterephthalate

dichlorodlphenyl dichloroethylene

dichloro-diphenyltrlchloroethane
0,0-dlethyl 0- (2-lsopropyl-4-methyl-
6-pyrlmidyl ) phosphorothloate
2,6-dlchloro-4-nitroanlllne

1,2,3.4, 10, 10-hexachloro-6,7-epoxy-

l,4,4a,5,6,7,8,8a-octahydro-l,4-endo-ea:o-

5,8-dlmethanonaphthalene

2.4-dlchloro-6-(p-chloroanllino) -s-
trlazine

1,2,3.4, 10,10-hexachloro-6,7-epoxy-

1.4,4a,5,6,7,8,8a-octahydro-l,4-en(io-en(Jo-

5.8-dimethanonaphthalene

O.O.O'.O'-tetraethyl-S-S'-methylenebls-

phosphorodithloate

ethyl hexyl ester ot 2.4-dlchlorophenoxy=

acetic acid

O-ethyl O-p-nltrophenyl phenylph06=

phonothloate

N-trichloromethylthlophthallmlde

l,4.5,6,7.8.8-heptachloro-3a,4,7,7a-tetra=

hydro-4,7-endo-methanoindene

1.4,5.6,7,8,8-heptachloro-2.3-epoxy-3a.4,7,

7a-tetrahydro-4.7-methanoindan

Same

Isobutyl ester ol 2.4-dlchlorophenoxyacetlc

acid

iso-ocytl ester of 2,4,5-trlchlorophenoxy=

acetic acid

iso-ocytl ester of 2,4-dlchlorophenoxyacetlc

acid

Isopropyl ester of 2,4,5-trlchlorophenoxy=

acetic acid

isopropyl ester of 2,4-dichlorophenoxyacetlc

acid

l,l-bis(p-chlorophenyl)-2,2.2-trichloro=

ethanol

y Isomer of benzene hexachloride

S-(1.2-bis(ethoxycarbonyl)ethyl]0,0-

dimethyl phosphorodithioate

l,l,l-trlchloro-2,2-bis(p-methoxyphenyl) =

ethane

0,0-dimethyl O-p-nitrophenyl phosphoro=

thioate

p-chiorophenylp-chlorobenzenesulfonate

0.0-diethyl-O-p-nitrophenyl phosphoro=

thioate

pentachloronltrobenzene

l,l-dichloro-2,2-bls(p-ethylphenyl) ethane

2-nltro-l,l-bls(p-chlorophenyl) propane

dimethyl 2.4,5-trichlorophenyl phosphoro=

thioate

terpene polychlorinates

l,2,4,5-tetrachloro-3-nitrobenzene

tetrachlorodlphenylethane

p-chlorophenyl-2,4.5-trlchlorophenylsulfone

l,3,4,5,6.7,8,8-octachloro-3a,4,7.7a-
tetrahydro-4,7-methanophthalan

Same

0,0-diethyl S- (ethylthlo) methyl phosphoro=

dithioate

6,7.8,9. 10,10-hexachloro-1.5,5a,6,9 ,9a-
hexahydro-6,9-methano-2.4,3-benzo=
dloxathiepin 3-oxide

octachlorocamphene

S-[[(p-chlorophenyl)thlo]inethyl]0,0-
diethyl phosphorodithioate

2-chloroallyl dlethyldlthiocarbamate

Vol. I, No. 1, June 1967

PESTICIDES
IN PEOPLE

CRITERIA FOR MONITORING

PESTICIDES IN PEOPLE INCLUDE

HIGH- AND LOW-EXPOSURE

CONDITIONS,

AGE, SEX DIFFERENCES

Anne R. Yobs'

As described here the program for assessing
pesticide residue levels in the Na.t ion's populace
is being carried out by the Pesticides Program,
National Communicable Disease Center, Bureau
of Disease Prevention and Environmental Con-
trol, Public Health Service, U. S. Department
of Health, Education, and. Welfare.

Monitoring Objective

The pui-pose of the human monitoring program
is to determine on a national scale the levels and
trends of certain more commonly used pesticide
chemicals, both in the general population and
in population segments where the occurrence of
more extensive exposure to pesticides is known
or suspected.

In the past, studies were made by various in-
vestigators assessing the concentration of
pesticides and or their metabolites in human
beings. These studies have provided a useful
body of infoiTnation concerning the relationship
of exposure to the human body's storage of
pesticides. However, such assessments were
limited in regard to the geographic coverage
of the sampling, the variety of conditions of
exposure, and the spectrum of pesticides in-
vestigated. They were also limited in the age
range, the sex distribution, and the size of the
sampled population. Probably the greatest
weakness in the earliei- studies was the
limited variety of body tissues tested. In
fact, this earlier work was essentially limited
to body fat. The present monitoring program
will provide statistically and epidemiologically
sound information for use in the evaluation of
the significance of man's total exposure to
pesticides.

■ Pesticide* ProKram. Nutiuniil Communicable Diieasc Center. Public
Hfiilih .Service. Bureau of Disease Pre\'ention and Environmental
Omlrol, U. .S. Di'i-nrtniont of Health. E<luratiun, and Welfare.
Atlanta, CeofKia 3033.1.

Progravi Design, Samples, and
Sampling Sites

Monitoring studies will be of two types, a
limited national survey of the general popula-
tion and an in-depth study of selected com-
munities in high-use areas.

In the sur\'ey being activated in calendar year
1967, tissues will be collected regularly from
the general population in 12 different areas of
the country. The number of specimens will be
relatively small at first to permit evaluation of
the approach and correction of any problem
areas. The program will be expanded later as
data indicate. Samples will be collected at post-
mortem examinations and from hospitalized
patients. Only body fat samples will be analyzed
at this time from post-mortem examinations.
Sample tissues from living patients will include
blood serum and adipose tissue removed in-
cidentally at surgery.

In-depth community studies, including moni-
toring, are in progress at these locations:

Arizona — Pivia and Maricopa Counties

California — State-wide

Colorado — Weld County

Florida — Dade County

Hawaii — Island of Oahii

Iowa — Johnson County

Louisiana — LaForche and Jefferson Parishes

Mich iga n — Berrien County

New Jersey — Monmouth County

Texas — Cameron and Hidalgo Counties

Washington — Wenatchee and Quincy Basins

Plans are under way for the initiation of addi-
tional studies in Idaho, South Carolina, Missis-
sippi, and Utah.

These studies are sampling three population
groups: (1) occupationally exposed workers,
(2) individuals not occupationally exposed but
known to be repeatedly exposed, and (3) the
general urban population. The occupationally
exposed group consists of one or more of the
following: agricultural applicators, workers in
pesticide formulating plants, pest control op-
erators, greenhouse workers, and aerial spray
pilots. Representatives in the repeatedly ex-
posed i)opulation are people living in environ-
ments where they may be expected to have
I'epeated nonoccupational exposure — these areas
are usually heavily agricultural. The general
urban population gi-oup represents individuals
whose exposures arc largely limited to pesticide
traces in food, water, and air plus occasional

Pesticides Monitoring Journal

household or garden use of pesticides. Since the
occupationally exposed group consists predom-
inantly of men, sampling of this group will
be restricted to adult males. However, the two
remaining groups will be equally divided be-
tween males and females with a reasonable age
spread.

Study procedures for each participant include
a detailed history of pesticide exposure and
usage, a complete medical history and physical
examination, and hematologic and biochemical
testing. Pesticide residue analyses will be per-
formed on urine and blood of all participants
and, when available, on body fat and other
tissues also. In addition, each study performs
an area pesticide-usage profile and analyzes
samples from the local environment. Tissues
taken by the Community Studies for general
population studies will be secured at post-
mortem examinations of accidental deaths.

Pesticides and Analytical Methods

Chlorinated hydrocarbon pesticides are known
to concentrate in animal and human fat and to
persist there for prolonged periods. Primary
emphasis will be given to detecting residues
from these chemical compounds and assessing
their levels. A serious problem is the lack of
suitable analytical procedures for detecting
different classes of pesticide chemicals in the
ranges expected in the general population and
applicable to several tissues. It is recognized
that human tissue samples may well contain
several pesticides of the same or other classes,
and the analysts will be expected to be alert to
identify them. As research progress may in-
dicate and require, and as technological develop-
ments permit, other tissues and other pesticides
may be added to this monitoring program.

Each Community Study has or is developing
laboratory competence in pesticide analysis and
the required hematological and biochemical test-
ing. They perform all testing for their own
locations, and some will perform the analytical
testing for the general monitoring program
using standardized procedures. All participat-
ing laboratories are required to maintain a
satisfactory standard of technical performance
as demonstrated in a quality control program
conducted by the Pesticide Research Laboratory
(Florida) of the Pesticides Program.

Standardization of Procedures

Guidelines and forms have been developed to
standardize the collection and recording of in-
formation, the sampling and handling of tissue
specimens, and laboratory test procedures. This
will permit the comparison of data among the
several participants in the monitoring program.
Information from the several study areas will
be combined to give an overall picture for the
Nation as a whole.

RESIDUES IN
FISH, WILDLIFE,
AND ESTUARIES

INDICATOR SPECIES NEAR TOP
OF FOOD CHAIN CHOSEN FOR
ASSESSMENT OF PESTICIDE BASE
LEVELS IN FISH AND WILDLIFE—
CLAMS, OYSTERS, AND SEDIMENT
IN ESTUARINE ENVIRONMENT

R. E. Johnson', T. C. Carver=, and E. H. Dustman'

Federal efforts to determine pesticide levels in
fish and wildlife are being carried out by the
Bureau of Sport Fisheries and Wildlife, U. S.
Department of the Interior. Monitoring estuar-
ine pesticide levels in clams, oysters, and sedi-
ments is a joint endeavor of the Bureau of
Commercial Fisheries, U. S. Department of the
Interior, and the Water Supply and Sea Re-
sources Program of the National Center for
Urban and Industrial Health, Public Health
Service, U. S. Department of Health, Educa-
tion, and Welfare.

Monitoring Objective

These monitoring programs will ascertain on a
national scale and independent of specific treat-
ments the levels and trends of certain pesticidal
chemicals in the bodies of selected forms of
animals and in estuarine sediments.

iBureau of Sport Fisheries and Wildlife, U. S. Department of the
Interior. Washincton. D. C. 20240.

-Patuxent Wildlife Research Center, Bureau of Commercial Fish-
eries. U. S. Department of the Interior, Laurel, Maryland 20810.

3Patuxent Wildlife Research Center, Bureau of Sport Fisheries and
Wildlife, U. S. Department of the Interior, Laurel, Maryland 20810.

Vol. I, No. 1, June 1967

National Pesticide Monitoring Site;

*

I

•

/

/

* •

\

^

Pesticides Monitoring Journal

•i^lr<^^ •

•4

FISH

t

*

PEOPLE

MARKET

SHELLFISH &

■

BASKET

ESTUARIES

SURVEY

«

SOIL

REGIONAL

#

FOOD COMMODITY •

WATER

SAMPLING

^t

WILDLIFE

r

%

I

Vol. I, No. 1, June 1967

-J

MONITORING FISH

Complete standardization of one fish species
for nationwide analysis is not possible; there-
fore, a minimum of three species has been des-
ignated for collection at each sampling site.
As with the wildlife forms, fish being sampled
are at or near the top of the food chain. These
include — listed in their order of preference,
depending upon availability at individual collec-
tion sites — carp, buffalo, black bass, channel
catfish, green sunfish, yellow perch, rainbow
trout, and squawfish.

Collection Sites and Savipling Frequency

Fifty locations have been chosen as collection
sites. These sites were selected to coincide
wherever possible with sampling locations for
monitoring pesticides in estuarine environments
and in fresh water. In some instances selection
of alternate locations was necessary to provide
for collection sites at points where appropriate
resident fish populations can be sampled, where
nets can be placed in streams with some perma-
nence and where commercial fishermen may be
relied upon to take desired fishes if State or
Federal crews are not available to do so. Some
collection sites are in the immediate vicinity
of other U. S. Fish and Wildlife Sei-\ace facil-
ities where manpower and equipment are read-
ily available.

Collections are taken twice a year, as close to
April and October as possible, at each of the
50 sampling locations; the amount of fish per
collection is from 1.5 to 2.5 lbs. Measurement of
pesticide levels at these times of the year in-
dicates body burdens immediately pi'ior to
spawning of some fish species and at a time of
maximum body fat content of neai-ly all species.
Sampling at these times also reflects levels
before and after major seasonal uses of
pesticides.

Sampling locations are listed by regional drain-
age systems:

Atlantic Coastal Drainage

Penobscot River, vicinity of Orono, Maine
Connecticut River, Windsor Locks, Connecticut
Hudson River, Poughkeepsie, New York
Delaware River, Camden, New Jersey
Susquehanna River, Conowingo Dam, Maryland
Potomac River, Little Falls, Maryland
Roanoke River, Weldon, North Caroliyia
Cape Fear River, Wilmington, North Carolina

Cooper River, Lake Moultrie or Marion, South

Carolina
Savannah River, above Savannah, Georgia
St. .Johns River, Welaka, Florida
St. Lucie Canal, Indiantown, Florida

Gulf Coastal Drainage

Apalachicola River, Jim Woodruff Dam, Florida
Tombigbee River, above Mobile, Alabama
Mississippi River, commercial fisheries. New

Orlcajtii, Louisiana
Rio Grande, above Brownsville, Texas

Great Lakes Drainage

Genessee River, near Avon, New York
Commercial fishery landings at:

Port Ontario, New York

Erie, Pennsylvania

Bay Port, Michigan

Port Washington, Wisconsin

Bayfield, Wisconsin

Mississippi River System

Kanawha River, Winficld, West Virginia
Ohio River, near Marietta, Ohio
Cumberland River, Clarksvillc, Tennessee
Illinois River, Beardstown, Illinois
Upper Mississippi River, Guttenberg, Iowa
Arkansas River, near Pine Bluff, Arkansas
Arkansas River, Keystone, Oklahoma
White River, near Dc Vails Bluff, Arkansas
Missouri River, Nebraska City, Nebraska
Missouri River, Garrisoti Dam, North Dakota
Missouri River, Fort Benton, Montana

Hudson Bay Drainage

Red River, near Noyes, Minnesota

Colorado River System

Green River, near Vernal, Utah
Colorado River, Imperial Dam, Arizona

Interior Basins

Lower Truckee River, Derby Dam, Nevada
Utah Lake, near Provo, Utah

California Strea^ns

Sacramento River, Sacramento, California

San Joaquin River, near Los Banos, California

Columbia System

Snake River, near Hagerman, Idaho
Snake River, Lcwiston Dain, Lewiston, Idaho
Salmon River, near Riggins, Idaho
Yakima River, near Prosser, Washington
Willamette River, above Oregoti City, Oregon
Columbia River, Bonneville Dam, Oregon

Pacific Coastal Streams

Klamath River, near Klamath River, California
Rogue River, near Grayits Pass, Oregon

Alaskan Strea7ns

Yukon-Tanana system, Fairbanks or Tanana,

Alaska
Kenai River, Soldatna, Alaska

10

Pesticides Monitoring Journal

Methods of Collecting, Preserving, and
Shipping Specimens

Fish are collected by seining, gill-netting, elec-
tric shocking, or by any other means which
insures that no extraneous chemicals are in-
troduced to complicate the analysis. Poisons
such as rotenone are not being used for collect-
ing samples, because they may interfere with
the analysis. Approximately 1 lb. of fish is taken
for each sample when the whole fish is to be
ground for analysis. When individual tissues
are to be analyzed, more than a pound of whole
fish may be required to furnish large enough
specimens of specific tissues.

Fish are wrapped in aluminum foil for pres-
ervation by freezing. Samples of fish are
wrapped separately to avoid contamination
between and among samples. Labels are made
for each sample on paper of a durable quality
and printed with soft black pencil rather than
ink to avoid smearing when wet. Each label
shows the name of the collector, specimen
number, species, sex and age if known, and
date and place of collection.

Frozen specimens are packed in a strong card-
board carton or drum with crumpled newspaper
or styrofoam for insulation and kept refriger-
ated with dry ice at the rate of 10 lbs. for 10-15
lbs. of fish. Samples are transported by air
freight or air express.

A telegram is sent to the receiving laboratory
before or at the time of shipment to prevent
delay in the pickup of specimens at destination
points. Also at the time of shipment, or soon
thereafter, a detailed list of the specimens is
forwarded to the laboratory.

MONITORING WILDLIFE

Since it is impossible to sample representatives
from each major group of animals occurring
in the United States, it is necessary to select,
at least for the time being, several species of
wildlife which occur reasonably close to the
top of a food chain. Later, as more is learned
about the significance of pesticide residues in
animal tissues, and as more is learned about
effects in various groups of animals, adjust-
ments can be made in the breadth and scope

of monitoring coverage in keeping with new
knowledge.

Several criteria are important in the selection
of forms for monitoring. The species selected
should not be extremely sensitive to chemicals
to be monitored. They should be geographically
well distributed, reasonably numerous, and easy
to collect. Residues in species close to the top of
a food chain also will reflect residues in organ-
isms at lower levels.

Species chosen for monitoring pesticides in
wildlife include the mallard duck, starling, and
the bald and golden eagles. The closely related
black duck is being substituted for the mallard
in States where adequate samples of the mallard
cannot be obtained.

DiLck Sampling

In cooperation with State agencies, the U. S.
Bureau of Sport Fisheries and Wildlife an-
nually collects thousands of waterfowl wings
of game species from all parts of the United
States. These are sent to several collection sites
throughout the country where they are housed
in freezers until they are examined by water-
fowl biologists to determine sex and age ratios.

The mallard is universally distributed in the
United States. It is a migrant form, moving
each spring into the northern United States
and Canada to breed. In the fall it moves south-
ward to winter. It is omnivorous and feeds in
both aquatic and terrestrial environments. Ap-
proximately fifty thousand mallard wings, and
where necessary black duck wings, are collected
annually during the hunting season in each of
the 48 conterminous States.

Through a process of systematic subsampling,
a series of wings are drawn from each State.
Approximately 12,500 wings annually are com-
posited into samples of 25 wings each for
analysis.

Starling Sampling

The starling is a ubiquitous bird which lends
itself well to sampling. Being omnivorous, it
feeds heavily on many kinds of insects and
fruits in spring, summer, and fall, and on crop
remnants and a miscellanea of other foods in
wnter months.

Vol. I, No. 1, June 1967

11

Beginning- in calendar year 1967, starlings will
be collected from various areas of the countiy
at two periods of the year, late summer and
winter. Three composite samples of 10 birds
each will be collected per seasonal sampling
period at each of 44 collection sites widely
distributed geographically. One set of samples
will be taken by trapping or shooting in August
when pesticide body burdens will reflect ap-
plications made during the growing season.
Another set will be taken in December or
January to assess body burdens during a period
of minimum pesticide usage throughout most
of the country.

Eagle Sampling

The golden and bald eagles currently are being
monitored by the U. S. Bureau of Sport Fish-
eries and Wildlife. Specimens found dead or
incapacitated and beyond recovery are sub-
mitted to Bureau laboratories for analysis. No
well-established sampling pattern is possible
with these forms, largely because of their pro-
tected status and their relatively low population
levels. They have been included in the pesticide
monitoring program because they are carnivo-
rous and at the top of food chains.

Methods of Handling Specimens

Wildlife specimens are handled and packaged
in the same manner as fish samples.

MONITORING ESTUARIES

The Federal estuarine pesticide monitoring
program is conducted jointly by the Bureau
of Commerical Fisheries of the U. S. Depart-
ment of the Interior and the Water Supply and
Sea Resources Program of the National Center
for Urban and Industrial Health, Public Health
Service, U. S. Department of Health, Education,
and Welfare.

Shellfish of interest are oysters and clams. These
filter-feeding pelecypod mollusks of commercial
value occur in all large estuarine systems in
the United States. They are particularly well
suited for pesticide monitoring because, as ses-
sile forms, they filter vast quantities of water.
They also are abundant and easily obtained.
Preliminary experimental work indicates that
oysters and clams will tolerate chlorinated

hydrocarbon pesticides and retain the residues
of these chemicals for extended periods follow-
ing exposure.

Sediment is of keen interest, because it is an
important part of the total aquatic environ-
ment. Pesticides adsorbed to soil particles
usually are chemically inactive although a slight
decrease in pH values can result in release of
the chemical from the soil particles. A similar
pH change is usually encountered in the upper
animal digestive tract. Many estuarine forms
are susceptible to this type of exposure.

While the basic orientation of this program is
toward commercial estuarine fisheries, other
areas of interest related to sediment monitoring
are recognized. For this program, sampling of
sediment is at the interface, which is the upper-
most portion of bottom sediments.

Sampling Sites

Samples for analyses are collected by agencies
at both the Federal and State level from estu-
arine systems and major river drainages con-
taining commercial quantities of shellfish. In
the interest of continuity, unifoiTn sampling
procedures are observed by each cooperating
organization. A total of 24 sampling locations
have been selected which serve as collection
sites for both shellfish and sediment. The samp-
ling point within each estuary is selected on the
basis of available hydrographic data, particu-
larly current patterns, and on the availability
of suitable shellfish populations.

Estuaries being studied, chosen on the basis of
water mass, are:

Penobscot Bay
Narragansett Bay
Long Islatid Sound
Pcconic Bay
Delaware Bay
Raritan Bay
Mid Chesapeake Bay
Lower Chesapeake Bay
Palmico Sound
Cape Fear River
Savannah River
Indian River

Tampa Bay
Apalacliicola Bay
Mobile Bay
Mississippi Sound
Lake Calcasieu
Barataria Bay
Galveston Bay
Humboldt Bay
San Francisco Bay
Willipa Harbor
Lower Puget Sound
Tillamook Bay

Sampling Frequency, Number of Samples

In order to evaluate adequately the annual trend
of pesticide residues in shellfish and estuarine

12

Pesticides Monitoring Journal

sediment, a minimum of three samples per year
are planned — in mid-March, mid-September,
and mid-November. However, existing pro-
grams of cooperating agencies are being incor-
porated in this program resulting in monthly
samplings at most sampling sites.

At each sampling station, 3 pools of 10 oysters
constitute the mollusk sample. Samples taken
3 times yearly from all collection sites will total
approximately 2,200 specimens per year.

Sampling Procedures

Oysters are the preferred mollusk at all moni-
toring sites. If commercial oysters are not avail-
able, any two species of local clams will be
substituted. Only adult oysters are taken. If
not endemic to the area, oysters are selected
that have a 1-year history in the water mass
from which they are taken. Persons collecting
samples are requested to furnish a complete
history of the oyster stock, a description of the
sampling site, and of the collection gear used.

Samples are preserved immediately and for-
warded without delay to appropriate regional
residue testing centers. Data accompanying
the samples include hydrographic observations
as well as the station data previously described.

Sampling procedures for sediment are the same
as those for mollusks with regard to frequency,
area, sample treatment, and shipment. Sample
origin is at the interface or uppermost layer
of the bottom sediment.

CHEMICALS MONITORED

IN FISH, WILDLIFE, AND

ESTAURIES

Of the many pesticidal chemicals now in use
and occurring in natural ecosystems, the follow-
ing are considered most important to fish, wild-
life, and estuaries: DDT (dichloro-diphenyltri-
chloroethane) and its metabolites, dieldrin (1,
2,3,4,10,10-hexachloro-6,7-epoxy-l,4,4a,5,6,7,8,
8a-octahydro-l,4-e?ido-exo-5,8-methanonaphtha-
lene), endrin (1, 2,3, 4,10, lO-hexachloro-6,7-
epoxy-l,4,4a,5,6,7,8,8a-octahydro-l,4-endo-enf/.o-
5,8-dimethanonaphthalene) , heptachlor (1,4,5,
6,7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-
enfZo-methanoindene) , heptachlor epoxide (1,4,
5,6,7,8,8-heptachloro-2,3, epoxy-3a, 4,7,7a-tetra-

hydro-4,7-methanoindan) , benzene hexachlor-
ide, lindane (gamma isomer of benzene hexa-
chloride), chlordane (1,2,3,5,6,7,8-octachloro-
2,3,3a,4,7,7a-hexahydro-4, 7-methanoindene) ,
and toxaphene (octachlorocamphene) . Each of
these compounds is included in the guide to
chemicals which has been established for the
overall national pesticide monitoring program
(p. 20) . This list may be modified as new chemi-
cals make their appearance or as new informa-
tion on present chemicals dictates.

CHEMICAL METHODOLOGY

The primary method of analysis will be the
latest methodology associated with gas chroma-
tographic techniques, with a randomized system
of cross checking with thin-layer chromato-
graphy.

The degree of sensitivity of residue determina-
tions will be no less than 1 x 10" (parts per
million) on a wet weight basis. Dry weights
of samples also will be obtained.

PESTICIDES

IN

WATER

NETWORK TO

MONITOR HTDROLOGIC

ENVIRONMENT

COVERS MAJOR DRAINAGE

RIVERS

by R. S. Greeni and S. K. Love=

This program for continuous surveillance of
pesticides in surface ■waters has been proposed
for joint operation by the Federal Water Pollu-
tion Control Administration and the Geological
Survey of the U. S. Department of the Intenor.
The proposal has been partially implemented.

1 Division of Pollution Surveillance. Federal Water Pollution Control
Administiation. U. S. Department of the Interior, Washington,
D C. 20242.

2 Quality of Water Branch, Geological Survey. U. S. Department of
the Interior, Washington, D. C. 20240.

Vol. I, No. 1, June 1967

13

Purpose and Objective

The purpose of this program is to provide con-
tinuing information on the overall extent of
pesticide contamination of the Nation's water
resources. The objective has been to develop
the minimum program that will enable an ade-
quate assessment of conditions. Within this ob-
jective, monitoring currently is confined to the
examination of surface waters in the major
drainage rivers of the United States through a
nationwide network of sampling locations. Over
a period of years, it is expected that data ob-
tained from this network will reflect impoi-tant
changes in pesticide levels in these rivers.

Design of River Network

Selection of sampling locations for a minimum
network to detect long-term or other significant
changes in pesticide levels in the water environ-
ment has required that consideration be given
primarily to area coverage, and only in a
secondary sense to the factors of pesticide use
or production.

Thus, the following criteria were used to select
locations on rivers for pesticide monitoring:
(1) locations to be near the mouths of rivers
that represent major river drainages through-
out the country; (2) river systems to be
sampled at other points where there is reason
to believe that a reasonable measure of pesti-
cide contamination cannot be obtained merely
by sampling at the mouth; (3) stations to be
at or near stream-gauging sites ; (4) considera-
tion to be given to locating sites where the
quality of river water is now being affected by
use of pesticides; (5) wherever practicable,
stations to be located at sites where other
kinds of water-quality data have been or are
being collected ; ( 6 ) wherever practicable, sta-
tions to be coordinated with suitably located
points from which historical data in the form
of carbon filter extracts are available.

Location of Sampling Sit^^s

Within the framework of the above criteria,
53 water sampling locations have been chosen
to provide preliminary information on the dis-
charge of pesticides in fresh water draining
from the conterminus United States. Monitor-
ing stations are located near the river mouths,

except on those streams discharging to tidal
waters. The latter stations are above areas of
salt-water intrusion.

Sampling points near the mouths of the follow-
ing streams were selected :

Connecticut River
Hudson River
Delaware River
S^isquehanna River
Potomac River
James River
Roanoke River
Cape Fear River
Pee Dee River
Santee River
Savannah River
Altamaha River
St. Jolins River
Siiwanee River
Apalachiocola River
Alabama River
Tombigbee River
Pearl River
Mississippi River
Atchafalaya River

Sabine River
Trinity River
Brazos River
Colorado River (Texas)
Nueces River
Rio Grande
Colorado River

(Arizona-California)
Los Angeles Aqueduct
San Joaquin River
Sacram.ento River
Klamath River
Columbia River
Ohio River
Illinois River
Missouri River
Arkansas River
Yakima River
Willamette River
Snake River

Streams selected for sampling at other locations
are as follows:

Upper Colorado River
(Arizona)

Middle Rio Grande

Upper Rio Grande

Middle Missouri River

Upper Missouri River

Middle Arkansas River

Upper Arkansas River

Middle Ohio River

St. Mary's River

(Michigan)
Lake Erie Outlet
St. Lawrence River

Red River of the North
(near Canadian Border)

Middle Mississippi River
(below Ohio and
Missouri Rivers)

Upper Mississippi River
(above Ohio and
Missouri Rivers)

Sample Collection Procedures, Frequency,
Preparation

Distinctive types of sampling programs are
required when monitoring the effects of a point
source of pollution as compared to monitoring
a stream at a site subject to a diffused source
of pollution. Because of its geographically broad
but minimal scope, this program is concerned
only with procedures for monitoring the latter
kind of sites.

14

Pesticides Monitoring Journal

The number and frequency of sampling is one
grab sample collected once a month at each
sampling site.

The following sampling procedures have been
prescribed:

All samples are to be collected in glass bottles.
Prior to collection, scrupulous cleansing of
sample containers is required. Chromic acid
cleaning solution or other suitable cleansing
agents are to be used, followed by several
rinsings with organic-free distilled water. Con-
tainers are to be further treated as necessary
to destroy remaining traces of organic matter ;
heat treating of containers at 300° C. has been
found satisfactory. Bottles are to be stoppered
immediately to prevent air-borne contamina-
tion. The sample must have no contact with
rubber, cork, and most plastics ; Teflon, how-
ever, will not contaminate the sample. Rubber
or cork stoppers may be used if wrapped care-
fully with a double layer of organic-free tin-
foil or aluminum foil, taking care to avoid rup-
ture of the foil cover when stoppering the
bottle.

The sample is to be collected in the prepared
glass bottle by lowering it in a weighted bottle
holder in a vertical section of the stream which
is representative of the stream cross section.
The bottle is to be lowered as nearly as possible
to the bottom of the stream and returned to
the surface so that all points in the vertical
section are represented in the sample.

Local conditions may prevent, or make unneces-
sary, fulfillment of all of the above conditions.
However, prior reconnaissance sampling at sev-
eral vertical sections of the stream may be re-
quired to determine degree of uniformity in
the cross section. If lack of complete mixing
(including floating of pesticides at or near the
surface) is suspected, notation of this effect
should be made.

It is important that the sample not be trans-
ferred from one container to another. Separate
containers must be used for determination of
any parameters that may be desired in addition
to pesticide levels. A sample tag or label pro-
viding appropriate identification is then com-
pleted and fii-mly affixed to each sample con-
tainer. Recorded information is also to include

river flow or stage, temperature, physical ap-
pearance of the water, or unusual physical
stream features.

Sample Shipment

Samples are to be shipped in suitable packing
cases to the laboratory for analysis as soon as
possible after collection, using parcel post, rail-
way express, air parcel post, or air express. It
is highly desirable that samples arrive at the
laboratory and extraction be commenced within
2 days after collection.

Sample Storage

When it is not possible for samples to be
analyzed within 1 week, the samples or their
extracts are to be stored in the dark and in a
cool place to retard growth of algae.

Extraction and Analysis of Compounds

Water analysis techniques involving instru-
mentation with electron capture and coulomet-
ric titration are sensitive in the parts per tril-
lion range, but interferences from organic
contaminants in the laboratory air, reagent
solutions, and sampling containers often pose
problems that must be overcome to utilize this
degree of sensitivity. Therefore, extreme care
must first be taken to maintain the sample as
pure as possible while handling and during the
analytical procedure.

Principal chemicals for identification include
lindane (gamma isomer of benzene hexachlor-
ide) ; heptachlor (1, 4,5,6,7,8, 8-heptachloro-3a,
4,7,7a-tetrahydro-4,7-endo-methanoindene) ;
heptachlor epoxide (l,4,5,6,7,8,8-heptachloro-2,
3,epoxy-3a,4,7,7a-tetrahydro-4,7-methanoin-
dan) ;aldrin (l,2,3,4,10,10-hexachloro-l,4,4a,5,
8,8a-hexahydro-l, 4-endo-€xo-5, 8-dimethano-
naphthalene) ; dieldrin (1,2,3,4,10,10-hexachlo-
ro-6,7-epoxy-l,4,4a,5,6,7,8,8a-octahydro-l, 4-
endo-e.TO-5, 8-dimethanonaphthalene) ; endrin
(l,2,3,4,10,10-hexachloro-6,7-epoxy-l,4,4a,5,6,
7,8, 8a-octahydro-l, 4-ewrfo-enrfo-5,8-dimethano-
naphthalene) ; o,p'-DDT, p/p'-BBT (dichloro-
diphenyltrichloroethane) ; and also the herbi-
cides, 2, 4-D (2, 4-dichlorophenoxyacetic acid)
and 2,4,5-T (2,4, 5-trichlorophenoxyacetic
acid). When other primary pesticides are

Vol. I, No. 1, June 1967

15

kno\\^l to be used in the drainage areas, these
arc to be souglit as will other comiiounds listed
in the standard guide for the national pesticide
monitoring program (p. 20).

Reporting Results

Results are to be reported in parts per trillion.
Because of varied and rapidly evolving analyti-
cal methodology, detailed records should be
maintained, giving the methods used for each
analysis. In addition to normal information
about dates of sampling and analysis and sam-
ple location, such additional details as the fol-
lowing are to be maintained : volume of sample
extracted, volume of extract injected, methods
used (i.e., electron capture, coulometric, infra-
red, etc.), and columns used (i.e., QF-1 fluorin-
ated silicone coated 5% by weight, etc.).

Beyond the Minimum Program

In addition to the overall national pesticide
monitoring program, more specific studies on
many aspects of pesticide contamination of
water resources are needed. These studies are
required to enable official and private agencies
to understand and predict the behavior of the
total water system and to discharge their re-
sponsibilities in evaluating, regulating, and
managing the Nation's water resources.

Detailed studies are especially necessary to
understand pesticide contamination in relation
to groundwater aquifers, sediment transport,
irrigation return flows and other drainage from
agricultural lands, near-ground precipitation,
industrial waste discharges, and other factors.

Although not a part of this national program,
the Great Lakes and key inland bodies of water
should be sampled witii proper attention being
paid to major lake currents. Because individual
sampling points in lakes are less valuable than
sampling points in rivers, lake sampling is best
collated with points of water use. This provides
useful information with resi ect to water use
and contributes to the general understanding
of pesticide contamination.

River sampling at low flows provides clues on
the pesticide content of ground water entering
the stream. Special studies of these complex
hydrologic situations may be required.

Observations in estuarine waters and bays
should be correlated with the sampling of major
river systems, thus providing a base for relating
contamination of the marine environment with
fresh-water contamination. Because of the com-
plexity of sampling within estuaries, the ap-
proach taken should be similar to that employed
in lake sampling; that is, most sampling points
should be associated with some beneficial use.

Public Water Supplies

Monitoring of pesticides in finished waters
of public water supplies does not yield signifi-
cantly more information about contamination
of the environment than would already be
known from monitoring of raw waters as-
sociated with the systems. Finished water
sampling, therefore, is not considered an essen-
tial part of this program. It is recognized, how-
ever, that a sufficient number of finished waters
should be examined to establish the general
level of pesticides in finished supplies, to show
the extent of removal of pesticides in the treat-
ment process, and to forestall potential problem
areas. Sampling of raw and finished water at
a few river locations that coincide with sources
of water supplies for large cities, for example,
will help delineate areas of significant pesticide
levels.

PESTICIDES

IN SOIL

NATIONAL SOIL MONITORING
PROGRAM STUDIES HIGH-,
LOW-, AND NONUSE AREAS

p. F. Sand», J. W. Gentryi, J. Bongberg^, and

M. S. Schochter^

Miich of the described soil monitoring program
is being carried out by the U. S. Department of
Agriculture as an established program. Other
j)hases of monitoring are to be conducted by the
USDA in cooperation with State and other
Federal agencies.

1 Plant Pest Control Division. AjjricuUural Research Service, U. S.

Department of Agriculture. Ilyattsvilie, Maryland 20TH2.
- Division of I'orest Post Control. Forest Service, U. S. Department

of .\^'riculLure. Arlin^rton. Virginia 22209.
3 Entomoloiyy Hesearch Division. Apricxiltural Research .Service,

U. S. Department of Apriculture. Beltsville. Maryland 20705.

16

Pesticides Monitoring Journal

Monitoring Objective

The objective of this program is to detei'mine
existing levels of pesticide residues in soils of
selected areas in the conterminous United States
and to detect any significant changes in these
levels. Soil monitoring sites were chosen where-
ever possible to coincide with sampling sites of
other agencies in the Federal pesticide moni-
toring network so that soil data may be coi'-
related with pesticide levels in other environ-
mental media.

Samples and Sampling Locations

In monitoring the effects of pesticides on the
agricultural environment, soils are being
studied extensively in areas of high pesticide
usage, as well as in areas of low use and non-
use.

Selection of high-use areas for monitoring pur-
poses was based on pesticide-use records ob-
tained from the literature and through local
surveys. In each case, responsible State agencies
were consulted concerning site selection before
the program was undertaken in any particular
area. Next, an intensive, direct survey was con-
ducted among landowners or commercial pes-
ticide applicators. Only those farms having ac-
curate pesticide-use records over a period of
years were chosen for the high-use studies.
Wherever possible, these records were compiled
by year for the past 10 years.

Intensive study areas are currently at single
locations in Alabama, Arizona, and in the Red
River Valley of North Dakota. Studies in these
areas were set up to run for a 3-year period
and will be phased out at the end of the 1967
season. Operations at Stuttgart, Arkansas;
Greenville, Mississippi ; and Utica, Mississippi,
were phased out in the fall of 1966 after a 3-
year sampling period was completed. Special
soil monitoring activities have been extended
to numerous other areas of the country where
pesticides are extensively employed in agricul-
ture. Five farms are included at each location.

These areas, and the principal crops they pro-
duce, include :

• Texas Lower Rio Grande Valley* — • cotton

• Dade County, Florida* ■ — vegetables

• Western North Carolina — apples

• Eastern South Carolina — vegetables

• Central Georgia — peaches

• Eastern Virginia — peanuts

• Monmoiitli County, New Jersey* — vegetables

• Adams County, Pennsylvania — fndts

• Berrien County, Michigan* — -fruits and vegetables

• Urbana, Illinois — com

• Western Iowa — com and soybeans

• Weld County, Colorado* — root crops

• Yuma County, Arizona — citrus fruits

• Wcnatchee Basin area, Washington* — fruits and
root crops (2 locations)

• Kern County, California — cotton, vegetables

• Tulelake area, California — small grains, root crops
* Soil monitoring sites coincide with U. S. Public

Health Service sites to monitor pesticides in
human beings.

Altogether, 23 study locations have been estab-
lished and maintained in high-use areas.

For comparative purposes, areas that have
received only one or two applications of pes-
ticides also have meaning in the study of resi-
dues in soils. These conditions were found in
forest areas where insect infestations have re-
quired only periodic control and on western
rangelands where insecticides have been used
periodically to control grasshoppers and
Momion crickets.

Areas in which there was no knowii previous
use of pesticides were included to indicate pos-
sible distribution of pesticides in soil environ-
ments not directly exposed to pesticide applica-
tion. Monitoring sites that were selected for
low- and nonuse phases of this program meet
the following specific criteria :

• Sites are on noncidtivated lands, and no site is in-
cluded which has been cultivated within the past
10 years;

• Some, but not all of the sites, are located near areas
chosen for high-2ise studies;

• Sites arc at least 1 mile from any known treated
areas and, where possible, include lands subject to
contamination from treated areas

• Sites are in locations remote from currcrit culti-
vation, e.g., ranges, forests, and wildlands.

In selecting these areas, cooperation again was
sought from Federal and State agencies re-
sponsible for management of public lands. Such
areas as parks, forests, and western rangelands
afforded preferable sites for these soil studies.
Records of land use were available from appro-
priate agencies. If suitable sites could not be
obtained through public land agencies, desirable
sites were determined by direct survey on a
local basis.

Vol. I, No. 1, June 1967

17

A total of 35 sites were selected for low- and
nomise area studies. Sites are distributed evenly
within each categmy to include forest areas,
arid rangcland, plains areas, and the eastern
hardwood region.

Sites selected for low-use studies, listed accord-
ing to their principal insect conti'ol efforts, are:

• Grasshopper control (dieldrin^, aldrirv') — Klamath
County, Oregon; Lincoln County, Idaho; Phillips
County, Montana; and Fremont County, Wyoming

• Japanese beetle control (dicldrin) — Pike County,
Kentucky

• Mosquito control (DDT''^) — Camp Drum, New York

• Forest insect control (DDT) — Davy Crockett Na-
tional Forest, Texas; Manistee-Huron National For-
est, Michigan; Thomas Jefferson National Forest,
Virginia; Chippewa National Forest, Minyiesota;
Coconino National Forest, Arizona; Lincoln Na-
tional Forest, New Mexico; Stanislaus National
Forest, California; Chcquamegon National Forest,
Wisconsin; Allegheny State Forest, Pennsylvania;
Eagle Lake State Forest, Maine; and Chattahoo-
chee National Forest, Georgia

Sites for nonuse studies include:

• Forest Service Lands — Pisgah National Forest,
North Carolina; Oconee National Forest, Georgia;
Francis Marion National Forest, Sozitli Carolina;
Cross Timbers Grasslands, Texas; Ozark National
Forest, Arkansas; Mark Twain National Forest,
Missouri; Buffalo Gap Grasslands, South Dakota;
Cache National Forest, Utah

• National Wildlife refiiges — Gulf Islands National
Wildlife Refuge, M ississippi ; Kofa National Wild-
life Refuge, Arizona; Okefenokce National Wild-
life Refuge, Georgia; Scney National Wildlife Ref-
uge, Michigan; Ravalli National Wildlife Refuge,
Montana; Ft. Niobrara National Wildlife Refuge,
Nebraska; San Andres National Wildlife Refuge,
New Mexico; Anahunr National Wildlife Refuge,
Texas; Missisquoi National Wildlife Refuge, Ver-
mont; Pea Island National Wildlife Refuge, North
Carolina

The Agricultural Research Sei-vice has devel-
oped a plan for expanding the national soil
monitoring progi'am. The proposed program
has been designed on a statistical basis for the
conterminous United States to provide informa-

■• 1.2J.4,I0,10-hexachloro-6,7-epoxy-1.4.4u,5,6.7,8.8a-octahydro-l

4-cndo-cio-5.8-dlmethanoniiphthalenc.
■■• 1.2,3,4. 10,10-hfxachloro-l. 4,4a, 5,8.8a-hcxahydro-1.4-endo-Pio-5

8-<llnuthanonaphthalfiuv
o dlchloru-dlphenyltrlchlorocthanc.

lion that will pinpoint major trouble areas
which then will require additional monitoring.
The objectives of the program are:

1. To establish the level of pesticide residues m soils
in reference to major land-use areas in the United
States.

2. To continue sampling the saine sites over a period

of tijne to provide information on rates of change
of pesticide residue levels in soils.

It is planned to initiate this program in fiscal
year 1968. Soil will be collected from approxi-
mately 15,000 sites over the conteraiinous
United States during a 4-year period.

Sampling Frequency, Number of Samples

Under the present program : samples are col-
lected once a year in high-use areas after sea-
sonal control measures. Approximately 2,600
soil samples were collected annually when all
23 sites were being sampled.

One sampling per year also is made in each
of the 35 low- and nonuse monitoring areas.
Ten samples are collected for each site, totaling
350 samples per year.

About 2,950 soil samples have been collected
annually for all phases of the soil monitoring
program.

Sample Collection Procedures

Each of the large-scale study areas in Ala-
bama, Arizona, and North Dakota contains ap-
proximately 1 square mile (610 acres) of agri-
cultural land. Each area is divided for soil
sampling purposes into 12 to 15 blocks of ap-
proximately 35-50 acres each. Collection proce-
dures involve both block and plot sampling.

• Block Sampling — Three samples are taken per
block at each sampling. Each sample consists of a
cotnposite of one core per acre per block. Cores are
spaced as equally as possible throughout the block
and arc taken both from the row and between rows
in cidtivated crops.

• Plot Sauipling — Intensive sampling of 20-acrc
plots witliin certain designed blocks is made to ob-
tain more precise data on accumulation or deple-
tion of pesticide residues, and to develop data on
rate of movement of pesticides with water.

The 20-acre plot is divided into 1-acre sections,
each to be sampled and analyzed separately.
One i-eprosentative sample consisting of 50

18

Pesticides Monitoring Journal

cores is taken from each acre. The location of
the intensive study plot within a selected block
is determined by watershed surveys. Where
possible, samples of each crop grown within a
study area are analyzed for pesticide residues
as a part of this study.

In the special soil studies in the high-use areas,
one representative field of 20 acres or more on
each of the five study farms is chosen for samp-
ling. Five 1-acre plots are laid out in this field
and samples collected on a stratified random
basis throughout the plot. Establishment of
these plots is made in reference to the field's
topography.

Ten 1-acre plots are laid out in each of the
low- and non-use areas. Fifty cores are collected
on a sti'atified random basis as in the high-use
area plots.

A uniform procedure for taking the cores, com-
positing the sample, and general handling of
the sample has been developed. Soil is sampled
to a depth of 3 inches with a corer 2 inches in
diameter. All cores contain the surface cover of
the soil, e.g., debris, sod, leaves, or any other
material which penetrates through normal
sampling. In forest areas of heavy duff, samples
are taken in spots where cover is liglitest. Cores
are collected in a large container, such as a 5-
gallon pail, and the combined cores are passed
twice through a i4.-inch screen to facilitate mix-
ing. Stones, roots, twigs, grass, etc., that do
not pass through the screen are discarded;
however, lumps of soil are forced through the
screen. A Y^S^^^on container is filled with the
mixed, screened soil and sealed with an air-
tight lid. The container then is labeled witli
sample number and date. A sample data sheet,
in an envelope, is fastened securely with tape
to the outside of the container. Equipment is
thoroughly cleaned after each sample collection,
and other measures are taken to guard against
contamination in all phases of the operation.

Pesticides and Analytical Methods

Analyses are performed to detect and identify
as many pesticides and important degradation
or metabolic products as possil^le. A general
guide for pesticides to be identified is that de-
veloped for the national pesticide monitoring
program (p. 20). This is augmented by

such knowledge and records as can be obtained
of pesticides used for agriculture, control of
forest pests, and for any other uses.

In the laboratory, subsamples of soil are used
for analysis; the remainder of each sample is
then stored until it is determined to be of no
further use. The latest and most sensitive
methods of analysis are employed, including
various procedures based on gas chromato-
graphy and paper and thin-layer chromato-
graphy. In doubtful cases, infrared spectropho-
tometry or colorimetric analyses also are
employed if a sufficient amount of pesticide is
present to peiTnit using these techniques.

In addition to other sources, the Guide to the
Analysis of Pesticide Residues (H. P. Burch-
field, 1965, Supt. of Docs., U. S. Government
Printing Office, Washington, D. C), is used to
select appropriate analytical procedures. Sample
sizes and sensitivity of the methods employed
are sufficient to permit reasonable detection
without making the analytical method unduly
cumbersome or complicated.

Factors which affect the sensitivity of the analy-
tical methods include:

1) sample size: 2) efficiency of extraction; 3) efficiency
of cleanup procedure (solvent partitions, column chrom-
atography, etc.); A) background due to naturally occur-
ring interferences; 5) interference from instrument
noise or fluctuations ; 6) interferences from solvents and
reagents; 7) cross interference of one pesticide (or its
degradation and metabolic prodxicts) with another; 8)
sensitivity of the final detection step, such as gas chrom-
atography irith its vaj-ious attached detectors as well as
spectrophotometry in the visible, ultraviolet, or infrared
regions, etc.

Importantly, sensitivities of the analytical
method may vary from one insecticide to an-
other, and even for the same insecticide from
one soil type to another. Sandy soils with low
organic content usually are less troublesome
to analyze than muck soil with high organic
content. Due consideration also is given to sig-
nificant pesticide degradation and metabolic
jiroducts to the extent that suitable analytical
procedures are available.

Reporting Results

Results of analyses are expressed in parts per
million on a diy-weight basis, and where pos-
sible in pounds per 3-inch acre.

Vol. I, No. 1, June 1967

19

CHEMICALS MONITORING GUIDE FOR

NATIONAL PESTICIDE MONITORING

PROGRAM'

Milton S. Schechter2
The purpose of this guide is to focus attention
on certain pesticides of special concern to the
national pesticide monitoring program. Two
lists of chemicals are presented as an aid to
participating Federal agencies in designating
pesticides to be identified in their initial moni-
toring studies. The primary listing contains
chemicals believed at present to be of most
interest because of their (1) extent and/ or
volume of usage; (2) degree of hazard to man,
fish, and wildlife; and (3) degree of persistence.
Pesticides on the secondary list are considered
to be of lesser importance or interest at
present. Both are minimal listings in keeping
with the minimum scope of the national pesti-
cide monitoring program; however, these lists
are not to be considered exclusive. All identifi-
able pesticides found in significant quantities
should be reported. This includes metabolic
and/ or breakdown products which are pesticidal
or toxic.

Not all of the pesticides listed, of course, can or
should be determined in all samples. Available
information on the use patterns of pesticides in
the areas \vliere samples are taken should be
used as a guide and consideration given to
possible movement of pesticides in the air
(drift), water (run-off), or soil (percolation).

These lists may be revised periodically to allow
for addition or deletion of pesticides according
to changes in their use, introduction of new
pesticide chemicals, and advances in analytical
methodology.

Because of difliculties involved in screening
samples for a multiplicity of pesticides and
their important metabolites and degradation
liroducts, care should be used not only in the
sampling and quantitative aspects of monitor-
ing studies but especially in the identification
aspects in order to assure reliability of reported
results.

20

' This Kuiiie was (Irnwn up by a proup of representatives (with
Milton S. Scliechtcr as chairman) from the U. S. Departments
of Agriculture, Defense, the Interior, and Health, Education,
and Welfare, under the sponsorship of the Subcommittee on
Pesticide Monitoring of the Federal Committee on Pest
Control.

- Entomology Research Division, Agricultural Research Service,
U. S. Department of Agriculture, BeltsvlUe, Maryland 20V05.

Pesticides Monitoring Journal

Primary List of Chemicals for Monitoring-i

Mercury-containing
pesticides (inor-

Common or Trade Name

Chemical Name

ganic and organic)

Aldrin

not less than 95% of 1,2,3,4, 10, 10-hexa=^

chloro-l,4,4a.5,8,8a-hexahydro-1.4-endo-

Mettioxychlor

l.l,I-trlchloro-2,2-bls(p-methoxyphenyl) =

eio-5,8-dimethanonaphthalene

ethane; technical methoxychlor contains
some o,p'-lsomer also

Amitrole

3 -amlno-s-triazole

Methyl parathion

0,0-dlmethyl 0-p-nitrophenyl phosphoro-

Arsenic-containing

thioate

pesticides (inor-

•

ganic and organic)

Mirex

dodecachlorooctahydro-1.3,4-metheno-
2H-cyclobuta[cd]pentalene

Azinptiosmetliyl

0,0-dlmethyl phosphorodlthloate S-ester

(Guthion®)

with 3-(mercaptomethyl) -1,2,3-

Parathion

0,0-dlethyl 0-p-nltrophenyI phosphoro

benzotrlazln-4 (3if ) -one

thloate

Benzene Inexa-

1 .2,3,4,5,6-hexachlorocyclohexane, consist-

clnloride (BHC)

ing of several Isomers and containing a

Silvex (including

2- (2.4, 5-trlchlorophenoxy) propionic acid

specified percentage of gamma Isomer

salts, esters, and
other derivatives)

Chlordane

l,2.4,5,6,7,8,8-octachloro-3a,4,7,7a-

tetrahydro-4,7-methanolndan; at least 60%

Strobane®

terpene polychlorlnates containing 65%

of l,2,4,5,6,7,8,8-octachloro-2,3,3a,4,7.

chlorine

7a-hexahydro-4.7-methanolndene and not

over 40% of related compounds

2,4, 5-T (including

2,4.5-trlchlorophenoxyacetlc acid

2,4-D (including
sales, esters, and

2,4-dichIorophenoxyacetlc acid

salts, esters, and
other derivatives)

other derivatives)

TDE (DDD) (includ-

l,l-dlchloro-2.2-bls(p-chlorophenyl) =

DDT (including its

l,l,l-trichIoro-2,2-bls(p-chlorophenyl) =

ing its isomers

ethane; technical TDE contains some

isomers and de-

ethane; technical DDT consists of a mix-

and dehydro-

o.p'-isomer also

hydroctilorination

ture of the p.p'-isomer and the o,p'-lsomer

chlorination

products)

(in a ratio of about 3 or 4 to 1)

products)

Dieldrin

not less than 85%; of 1.2,3,4, 10, 10-hexa =

Toxaphene

chlorinated camphene containing

chloro-6.7-epoxy-l,4,4a.5,6,7,8,8a-octa=

67-69% chlorine

hydro-l,4-en(Jo-eio-5,8-dimethanano=

Dithiocarbamate
pesticides:

naphthalene

Secondary List of Chemicals for Monitoring

IVlaneb;

( ethylenebls ( dlthlocarbamato) ]manga-

Demeton (Systox®)

mixture of 0,0-diethyl S(and 0) -

Ferbam;

nese;

trls ( dlmethyldithlocarbamato ) iron :

[2-(ethylthio) ethyl] phosphorothioates

Zineb;

[ethylenebls (dlthlocarbamato) Jzlnc;

etc.

Disulfoton

0.0-diethyl S-[2-(ethyIthlo) ethyl]

(Di-Syston®)

phosphorodlthloate

Endrin

1,2,3,4, 10, 10-hexachloro-6,7-epoxy-

l,4,4a,5,6,7,8,8a-octahydro-l,4-en(Jo-

Endosulfan

1.4.5,6,7,7-hexachloro-5-norbomene-2,3-

endo-5,8-dlmethanonaphthalene

(Thiodan® )

dimethanol cyclic sulfite

Heptachlor

l,4,5,6,7,8,8-heptachloro-3a,4,7,7a-

Inorganic Bromide

tetrahydro-4,7-methanolndene

from bromine-
containing

Heptachlor epoxide

1.4,5,6,7 ,8,8-heptachloro-2,3-epoxy-
3a,4,7,7a-tetrahydro-4,7-methanolndan

pesticides
Triazine-type

Lindane

1,2,3,4, 5,6-hexachlorocyclohexane,

herbicides;

gamma Isomer of not less than 99% purity

Atrazine;

2-chloro-4-(ethylamino)-6-
(Isopropylamlno) -s-triazine;

Malathion

diethyl mercaptosucclnate S-ester with

Simazine;

2-chloro-4,6-bls(ethylamlno) -s-trlazlne;

0,0-dimethyl phosphorodlthloate

etc.

^ The first chemical name given aft^r the common or trade

mark name Is according to Chemical Abstracts. The second

chemical name, if one Is given, is taken from "A List of

Insect Control Chemicals to be Used In Entomology Research

Note: A chemical name occupying two lines separated by an

Division Manuscripts" compiled by E. M. Osborne and Ruth L.

Busbev. Pesticide Chemicals Research Branch. Entomology

equall=) sign is joined together as one word, without

Research Division, ARS, USDA, June 1966.

the equal sign, if written on one line.

Vol. I, No. 1, June 1967

21

22

Information For Contributors

The Pesticides Monitoring Journal welcomes from all
sources qualified data and interpretive information
which contributes 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 cir-
culation is maintained for persons with related in-
terests, notably those in the agricultural, chemical
manufacturing, 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 references. Accuracy, reliability, and limitations of
the sampling 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 in-
terlaboratory checks. The procedure employed should be
referenced or outlined in brief form, and crucial points
or modifications should be noted. Check or control sam-
ples should be employed where possible, and the sen-
sitivity 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 conformance
to the Style Manual for Biological Journals, American
Institute of Biological Sciences, Washington, D. C,
and/or the Style Manual of the United States Govern-
ment Printing Office, and an abstract (not to exceed
two hundred words) should accompany each manuscript
submitted.

Pesticides ordinarily should be identified by common or
generic names approved by national scientific societies.
The first reference to a particular pesticide should be
followed by the chemical or scientific name in paren-
theses— 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 information by any specific Federal agency
is not intended or to be implied. Authors of accepted
manuscripts will receive edited tjTDescripts 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, notation of such
should be provided.

Correspondence on editorial and circulation matters
should be addressed to: Editorial Manager, Pesticides
Monitoring Journal, Pesticides Program, National Com-
municable Disease Center, Atlanta, Georgia 30333.

Pesticides Monitoring Journal

The Pesticides Monitoring Journal is published quarterly under the auspices of the Federal
Committee on Pest Control and its Subcommittee on Pesticide Monitoring as a source of
information on pesticide levels relative to man and his environment.

The parent committee is composed of representatives of the U. S. Departments of Agriculture,
Defense, the Interior, and Health, Education, and Welfare.

The Pesticide Montoring Subcommittee consists of representatives of the Agricultural Research
Service, Consumer and Marketing Service, Federal Extension Service, Forest Service, Depart-
ment of Defense, Fish and Wildlife Service, Geological Survey, Federal Water Pollution
Control Administration. Food and Drug Administration. Public Health Service, and the
Tennessee Valley Authority.

Responsibility for publishing the Pesticides Monitoring Journal has been accepted by the
Pesticides Program of the Public Health Service.

Pesticide monitoring activities of the Federal Government, particularly in those agencies repre-
sented on the Pesticide Monitoring Subcommittee which participate in operation of the national
pesticides monitoring network, are expected to be 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 nongovernment research
institutions, both within and without the United States. 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 Subcommittee. Authors are given the benefit of review comments prior to
publication.

Editorial Advisory Board members are:

Reo E. Duggan. Food and Drug Administration, Chairman

Andrew W. Breidenbach, Public Healtli Service

Anne R. Yobs, Public Health Service

James B. DeWitt, Fish and Wildlife Service

S. Kenneth Love. Geological Survey

Milton S. Schechter, Agricultural Research Service

Paul F. Sand, Agricultural Research Service

Trade names appearing in the Pesticides Monitoring Journal are for identification only and do
not represent endorsement by any Federal agency.

Address correspondence to:

Mrs. Sylvia P. O'Rear

Editorial Manager
PESTICIDES MONITORING JOURNAL

Pesticides Program

National Communicable Disease Center

Atlanta, Georgia 30333

CONTENTS

Volume 1 September 1967 Number 2

Page

EDITORIAL 1

R. E. Duggan

RESIDUES IN FOOD AND FEED

Pesticide residues in lolcd diet samples (II) 2

R. E. Duggan, H. C. Barry, and

L. Y. Johnson
Chlorinated hydrocarbon pesticide residues
in or on alfalfa grown in soil with a previous
history of aldrin and hcptachlor application 13

R. J. Moiibry, G. R. Myrdal, and

H. P. Jensen

PESTICIDES IN PEOPLE

Storage of DDT in the people of Israel 15

M. Wassermann, Dora Wassermann,
L. Zellermayer, and M. Gon

RESIDUES IN FISH, WILDLIFE, AND
ESTUARIES

Insecticide concentrations in wildlife at

Presidio, Texas 21

Dudley D. Culley and

Howard G. Applegate
An evaluation of the effects of the Aedes
aegypti Eradication Program on wildlife in
south Florida - 29

Philip N. Lehner, Thomas O.

Boswell, and Frank Copeland
Pesticides in hatchery trout — differences
between species and residue levels occurring
in commercial fish food —^ 35

H. Cole, A. Bradford, D. Barry,

P. Baumgamer, and D. E. H. Frear

PESTICIDES IN WATER

Pesticides in selected western streams —

a contribution to the national program 38

F. Brown and Y. A. Nishioka
Persistence and movement of paralhion
in irrigation waters 47

C. W. Miller, W. E. Tomlinson,

and R. L. Norgren

GENERAL

Systemic activity of Zectran, Matacil,

and Bidrin injected into conifer trunks 49

John F. Larson, G. R. Pieper, and

H. C. Ratsch
Problems in im'nitoring DDT ami its
metabolites in the environment 54

Donald A. Spencer

EDITORIAL

PUBLICATION of original papers and data on pesti-
cide monitoring begins with this issue. We have been
pleased by the submission of many excellent articles
and by the gracious response of the authors to the
comments and suggestions of the review board.

Since the original announcement of plans for publica-
tion, there have been numerous questions concerning
the type and scope of reports acceptable to the
Journal. We are hesitant to define limits of subject
matter acceptability except in the rather broad terms
of significant information and data on the levels of
pesticides in all portions of the environment.

Monitoring, as used in the title of this Journal, may
be defined as the systematic recording of information
relating to the distribution and movement of pesticides
and their residues in the environment. Reports con-
cerned with this broad field are within the purview of
the Journal. There is no publication specializing in
baseline data and the everchanging levels of pesticides
in influential and significant environmental factors. The
policy of this Journal is to publish reliable information
on this broad subject in such a form as to be readily
used, compared, interpreted, and correlated by scientists
and others concerned with pesticides and their residues.

The National Pesticide Monitoring Program can be
only a frame to support and maintain a continuity with
other monitoring activities. Data derived from many
sources other than the National Pesticide Monitoring
Program described in the first issue are needed for a
more complete understanding of the effects of pesticides
on the environment. Informative articles may be pre-
pared from certain methodology and research projects
where the monitoring part of the investigation is second-

ary, and the monitormg data cannot be properly re-
ported elsewliere in sufficient detail for use by others.
Also, surveillance data from water, forest, or wildlife
conservation programs and that obtained during the en-
forcement of tolerances for residues should be valuable
sources of information. The Editorial Advisory Board
wishes to renew the invitation for manuscripts concerned
with monitoring pesticides from all sources.

There have been a few expressions of opinion that the
requirements for information on confirmatory analyses,
recovery experiments, sensitivity, and other items men-
tioned in Information to Contributors are too stringent.
We do not have requirements beyond sound and reason-
able criteria for scientific investigations. The uses for
monitoring data are extensive, and information in suffi-
cient depth is needed to permit the reader to make an
independent judgment concerning the data and their
range of usefulness. The editorial policy of the Journal
is to fulfill this need insofar as practical in order that
the readers will find the Journal a dependable, accurate,
and useful source of information. For example, we have
suggested that lengthy tabular data be presented in a
summarized form. The participants in the National Pesti-
cides Monitoring Program have agreed to respond to
requests from readers for more detailed data. We hope
other contributors will be equally responsive to such
requests. In future issues, we plan to devote this page to
discussions by members of the Editorial Advisory Board
on special requirements and important aspects of de-
finitive monitoring programs.

R. E. Duggan

Chairman, Editorial Advisory Board

Vol. 1, No. 2, September 1967

1

RESIDUES IN FOOD AND FEED

Pesticide Residues in Total Diet Samples (II)

R. E. Duggan', H. C. Barry-', and L. Y. Johnson'

ABSTRACT

Pesticide residue levels detected in ready-to-eat foods re-
mained at low levels during the second year of the total diet
study. Food samples were taken in 36 different markets
which were located in 25 different cities representing five
geographical regions. Averages and ranges of pesticides
found are reported for each year by region and food class.

THE study of pesticide residues in ready-to-eat foods
which was conducted by the Food and Drug Adminis-
tration from June 1964 through April 1965 was de-
scribed in an earlier report (/). The sampling, composi-
ting, and analytical schemes were given in detail in the
Food and Feed section of the initial issue of the Pesti-
cides Monitoring Journal which described the National
Pesticide Monitoring Program (2). This paper presents
data collected from June 1965 through April 1966.
More complete data for both periods are included in
tabular form.

The study was expanded beginning in August 1965 to
include samples from Minneapolis, Minn., and Balti-
more, Md. Sampling was not confined to the five metro-
politan areas; some samples were collected from smaller
cities.

Two significant procedural changes were made in Au-
gust 1965. One was the use of an improved analytical
procedure in which gas-liquid chromatography (3).
the thermionic detector, and the isolation procedure for
chlorinated organic compounds (4) were used to detect
organic phosphorus compounds. This procedure deter-
mines ethion, ronnel, carbophcnothion, malathion, di-
azinon. methyl parathion, and parathion at a sensitivity
level of 0.05 ppm. Thin layer chromatography (5) was
used for confirmatory analyses. The second procedural
change was the analysis of carbaryl by a thin layer
procedure (6).

' OITice of Associaic Commissioner for Compliance, Food and Drue
AdmmiMr.ition, II. S. Department of Health, Education, and Wel-
fare. Washintlon, D C. 20204.

' New Orleans District, Food and Drug Administration, New Orleans,
Louisiana 701. '0.

• Cincinnati District, Food and Drug Administration, Cincinnati,
Ohio 45210.

All methods used in these studies are described in the
FDA Pesticide Analytical Manual (1965), and may in-
clude refinements not described in the basic references
listed above. Recoveries of specific pesticide chemicals
vary within product classes, usually within a range of
85 9f to 115% at these levels. No correction was made
for recovery.

Quantitative values reported for chlorinated organic
compounds were obtained by electron capture gas-
liquid chromatography and confirmed by thin layer
chromatography, microcoulometric gas-liquid chroma-
tography, or both.

Results

Thirty-three different residues were found in the
samples in 1966. The frequency of the residues and
the ranges of their amounts are shown in Table 1. The
most common residues, maximum levels of these resi-
dues, and residues reported less frequently are discussed
for each food class.

DAIRY PRODUCTS: Thirteen chlorinated organic
pesticides in varying combinations were detected in 21
of 22 composites. The most common and their maxi-
mum values on a fat basis were DDE (0.58 ppm), DDT
(0.19 ppm), dieldrin (0.06 ppm), heptachlor epoxide
(0.077 ppm). and TDE (0.065 ppm). Aldrin, BHC,
lindane, methoxychlor, 2,4,5-TP, 2-4-DB, and PCP
and MCP were also present. Bromides were found (0.5
ppm to 21.4 ppm) in 25 of 28 composites.

MEAT, FISH, AND POULTRY: Nine chlorinated
organic pesticides were present in varying quantities in
22 of 26 composites. DDT, DDE. TDE, heptachlor
epoxide, dieldrin, and BHC were the most common,
with maximum values of 1.39 ppm, 1.0 ppm, 0.78 ppm,
0.29 ppm, 0.20 ppm, and 0.20 ppm, respectively, on a
fat basis. Lindane, tetradifon, and PCP were also present.
Bromides were detected (0.5 ppm to 44 ppm) in 23 of
28 composites, and arsenic (0.1 ppm to 0.5 ppm As-jO-,)
in 5 of 28 composites. Diazinon (0.051 ppm) and
ronnel (0.011) were found in 1 composite each.

Pesticides Monitoring Journal

TABLE 1. — Number of composites where pesticide residues were found and ranges in the

amounts (June 1965 - April 1966}

Pesticide

No.

COMPOSITES

WITH

RESIDUE

No. OF POSITIVE

COMPOSITES WITH

RESIDUES BELOW

SENSITIVITY LEVEL •

Range at and
above sensi-
tivity level

(PPM)

BROMIDES

DDT

1 , 1 , 1-trichloro-2,2-bis(p-chlorophenyI ) ethane

DDE

l,l-dichloro-2,2-bis(p-chloroptienyl) ethylene

TDE

l,l-dichIoro-2,2-bis(p-chlorophenyl) ethane

DIELDRIN

not less than SSrh of I,2,3,4,10,I0-hexachloro-6,7-epoxy-l,4,4a.5.6.7,8,8a-octahydro-
l,4-endo-f.vo-5,8-dimethanonaphthalene

LINDANE

1,2,3,4,5.6-hexachlorocyclohexane, 99'}r or more gamma isomer

HEPTACHLOR EPOXIDE

l,4,5.6,7,8,8-heptachIoro-2,3-epoxy-3a,4,7.7a-tetrahydro-4,7-methanoindan

BHC

1,2,3.4,5,6-hexachlorocyclohexane, mixed isomers

MALATHION

diethyl mercaptosuccinate, 5-ester with f).0-dimethyl phosphorodithioate

ALDRIN

not less than 95% of I,2,3,4,10,10-hexachloro-1.4,4a,5.8.8a-hexahydro-1.4-ciirfo-fio-
5,8-dimethanonaphthaIene
KELTHANE®

4, 4'-dichloro-a-(trichloromethyl) benzhydrol

PCP

pentachlorophenol

ARSENIC (AsiOa)
2,4-D

2,4-dichIorophenoxyacetic acid
DIAZINON

0.0-diethyl 0-(2-isopropyl-6-methyl-4-pyrimidinyl) phosphorothioate

CARBARYL

1-naphthyl methylcarbamate
ENDRIN

l,2.3,4,10,10-hcxachloro-6,7-epoxy-l,4,4a,5.6,7,8.8a-octahydro-l,4-eiirfo-fndo-
5,8-dimethanonaphthalene

ENDOSULFAN

6,7,8,9.10,10-hexachloro-l,5,5a.6,9.9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin

3-oxide
METHOXYCHLOR

1 , 1 , l-trichloro-2,2-bis ( p-methoxyphenyl ) ethane
MCP

4-chloro-2-methyl-phenoxyacetic acid
PERTHANE®

l,l-dichloro-2,2-bis(p-ethylphenyl) ethane
PARATHION

0,0-diethyl 0-p-nitrophenyl phosphorothioate
TOXAPHENE

chlorinated camphene containing 67% to 69% chlorine
RONNEL

0,0-dimethyl 0-2,4,5-trichlorophenyl phosphorothioate

CIPC

isopropl TS'-CS-chlorophenyl) carbamate
TETRADIFON

p-chlorophenyl 2,4,5-trichlorophenyl sulfone
2,4,5-TP

2-(2,4,5-trichlorophenoxy) propionic acid

TCNB

1,2,4, 5-tetrachloro-3-nitrobenzene
2,4-DB

4-(2,4-dichIorophenoxy) butyric acid

DACTHAL®

dimethyl ester of tetrachloroterephthalic acid

CHLORDANE

1,2,4,5,6,7,8, 8-octachloro-3a, 4,7,7a-tetrahydro-4,7-methanoindane

PCNB

pentachloronitrobenzene
ETHION

0,0,0' ,0'-tetraethyl S,S' methylene bisphosphorodithioate

244
119

119

83

75

49

42
21
16
13

12

10

10
9

9

8
6

0

7

32
13

22

21
9
3

13

3

0
5

0

3

8
5
0

0.5-117.0
O.0O4-1.39

0.003-1.00

0.003-0.78

0.003-0.20

0.003-0.080
0.004-0.29
0.008-0.20
0.053-0.15
0.005-0.070

0.013-0.21

0.036-0.31

0.1-0.5
0.051-0.10

0.051

0.2-0.4

0.004-0.052

0.006-0.016

0.004-0.073

0.039-0.58

0.007-0.057

0.089

0

0.048-0.38

3

0

0.20-0.36

0

0.011-0.076

0

0.018-0.029

1

0.37

2

2

0

0.37

0

0.005

1

Pesticide chemicals capable of being detected by the specified analytical methodology :
when they are present at concentrations below the sensitivity level.

be confirmed qualitatively but are not quantifiable

Vol. 1, No. 2, September 1967

GRAIN AND CEREAL PRODUCTS: Thirteen
chlorinated organic pesticides were found in 23 of 26
composites, with the most common being lindane, DDT,
and DDE at maximum values of 0.028 ppm, 0.024 ppm,
and 0.004 ppm, respectively. Other chlorinated organic
pesticides present were aldrin, BHC, dieldrin, heptachlor
epoxide, methoxychior, TDE, 2,4-DB, PCNB, Perthane,
and PCP. Bromides were present (1.1 ppm to 66.7
ppm) in 27 of 28 composites, and arsenic at 0.1
(As^.O;,) ppm in 1 composite. Eleven composites con-
tained detectable malathion with the maximum level
0.15 ppm. Diazinon and ronnel were also present.

POTATOES: Dieldrin and DDT at maximum values of
0.003 and 0.010 ppm, respectively, were the most com-
mon of 10 chlorinated organic pesticides found in 14
of 26 composites. Also, BHC, DDE, endrin, heptachlor
epoxide, lindane. TCNB, CIPC, and TDE were present.
Bromides were found (0.7 ppm to 68.5 ppm) in 20 of
28 composites, and parathion at 0.003 ppm in 1
composite.

LEAFY VEGETABLES: While DDT, DDE, and TDE
with maximum values of 0.048 ppm, 0.033 ppm, and
0.024 ppm. respectively, were the most common of 10
chlorinated organic pesticides found in 20 of 26 com-
posites, others were also detected. They were Dacthal,
dieldrin, lindane, endosulfan, toxaphene, 2,4-D, and
MCP. Bromides were found (0.6 ppm to 14.8 ppm) in
22 of 28 composites. Diazinon and parathion were de-
tected in 3 composites with maximum levels of 0.031
ppm and 0.089 ppm. respectively; malathion was found
in 1 composite.

LEGUME VEGETABLES: Ten of twenty-six com-
posites were found to contain seven chlorinated organic
pesticides. The most common were DDE and TDE,
with maximum values of 0.003 ppm and 0.064 ppm,
respectively; but DDT, aldrin, heptachlor epoxide, and
trace amounts of dieldrin and lindane were also de-
tected. Bromides were found (0.5 ppm to 14.5 ppm) in
22 of 28 composites.

ROOT VEGETABLES: The 6 chlorinated organic
pesticides found in 9 of 26 composites were DDE, dield-
rin (most common: maximum values of 0.01 1 ppm and
0.028 ppm, respectively), DDT, endrin, TCNB, and
TDE. Bromides were found (0.6 ppm to 17.0 ppm) in
21 of 28 composites, and arsenic (O.I ppm As-.O.)) was
found in 1 composite. Malathion (0.22 ppm) and
carbaryl (0.1 ppm) were also found in 1 composite.

GARDEN FRUITS: A total of 1 1 chlorinated organic
pesticide residues were found in 22 of 26 composites.
DDT. TDE. lindane, and DDE were most common,
with maximum levels of 0.17 ppm, 0.34 ppm, 0.012
ppm. and 0.064 ppm, respectively. The remainder were

dieldrin, aldrin, endrin, heptachlor epoxide, endosulfan,
toxaphene, and chlordanc. Bromides were found (0.5
ppm to 7.5 ppm) in 24 of 28 composites; arsenic (0.1
ppm As.O.) in 1 composite; and diazinon (0.005 ppm)
in 1 composite.

FRUITS: The most common of 1 1 chlorinated organic
pesticide residues found in 19 of 26 composites were
DDE. Kelthane, and DDT; their maximum levels were
0.043 ppm, 0.21 ppm, and 0.045 ppm, respectively. Less
frequently present were aldrin, Dacthal, dieldrin, Per-
thane, lindane, TDE, tetradifon, and endosulfan. Bro-
mides were present (0.7 ppm to 25.2 ppm) in 19 of 28
composites, carbaryl (maximum level 0.2 ppm) in 4
composites, and ethion (maximum level 0.019 ppm) in
I composite.

OILS, FATS AND SHORTENING: A total of 9
chlorinated organic pesticide residues were found in 17
of 26 composites. DDE, DDT, and TDE were the most
common, with maximum levels on a fat basis of 0.029
ppm. 0.038 ppm, and 0.12 ppm, respectively. Aldrin,
dieldrin, endrin, heptachlor epoxide, Perthane, and PCP
made up the other 6. Bromides were present (0.9 ppm
to 90.8 ppm) in 21 of 28 composites. The organophos-
phate, malathion (maximum level 0.18 ppm), was
found in 4 composites,

SUGARS AND ADJUNCTS: Of 7 chlorinated organic
pesticide residues found in 9 of 26 composites, 2,4-D, at
a maximum level of 0.1 ppm, was the most common,
DDE, DDT, dieldrin, MCP, and trace amounts of
lindane and heptachlor epoxide were also found. Bro-
mides were present (0.7 ppm to 1 17 ppm) in 25 of 28
composites, but arsenic was found (0.1 ppm As^O;;) in
only 1. Carbaryl was found in 2 composites at a maxi-
mum level of 0.2 ppm, and ronnel was found in 1 com-
posite in trace amounts.

BEVERAGES: A total of 3 chlorinated organic pesti-
cides were found in 2 of 26 composites. PCP (0.02
ppm) was found in 1 composite, and trace amounts of
lindane and heptachlor epoxide were also reported.
Bromides in concentrations ranging from 0.5 ppm to
13.7 ppm were found in 9 of 28 composites. Carbaryl
was found in 1 composite at 0.4 ppm.

Bromide residues were in excess of the quantitative
sensitivity limits established for the investigation in 258
of 336 composite samples. This incidence is 76.8%
and does not differ significantly from, the 1964-1965
results; however, there was a lower incidence of resi-
dues exceeding 25 ppm — 3.8% compared to 11.6%.
The presence of chlorinated organic pesticides was
confirmed in 168 of 312 composites examined (53.8%)
for this class of chemicals; this percentage of incidence
also is not significantly diflcrent from the 1964-1965

Pesticides Monitoring Journal

data. In contrast, the finding of residues of chlorophen-
oxy compounds in 24 composites was an increase over
the previous period. Carbaryl was found in 8 com-
posites— again a lower incidence than in the preceding
period. Organic phosphorus compounds were found in
27 composites; we attribute the finding of this class of
compound primarily to the improved analytical pro-
cedures used. Amitrole and dithiocarbamate residues
were not found at or above the prescribed sensitivity
limits.

Discussion

Levels of residues for the interval of this study remain
on the same order of magnitude as those reported in
the earlier studies. In addition, the frequency of resi-
dues encountered has not changed significantly as a
whole or within food classes. Residues of toxaphene,
Dacthal, endosulfan, 2,4.5-TP, diazinon, ronnel, mala-
thion, parathion, 2.4-DB, CIPC, and ethion have not
been reported previously in studies of foods ready for
consumption.

Data obtained during both periods of study are reported
in more detail in Table 2a where the findings are ar-
ranged by food class, region, and sampling period.
Period average, number of positive composite samples
and range of positive findings are given for those pesti-
cide residues commonly found. Similar information on
pesticide residues found infrequently is given in Table
2b. Trace amounts, < 0.001 ppm, were not in-
cluded in the averages. Where no average value is
given, results on individual composites are shown.

On the basis of these data, average daily pesticide in-
take from the diet was calculated and is reported else-
where (7). There was no statistically significant diff'er-
ence in the dietary intake between the two reporting
periods, and the calculated levels were not seen to be
approaching the acceptable daily intakes established for
certain pesticides chemicals by the World Health
Organization.

TABLE 2a. — Levels of Pesticide Residues Commonly Found — hy Food Class, Region, and Sampling Period

IT = Trace<0.001 ppm]

Pesticide

Boston
1965 1966

Kansas City
1965 1966

Los Angeles
1965 1966

Baltimore
1966

Minneapolis
1966

I. DAIRY PRODUCTS (8-13% fat) i
Residues in Parts Per Million — Fat Basis

DDT

Average

Positive Composites
Number

O.OIO

2

0.029
4

0.065
6

0.039
6

0.031
2

0.073
4

0.035
3

None

examined.

Range

0.011-0.048

T(a)-0.173

0.019-0.153

0.014-0.112

0.060-0.080

0.046-0.190

0.021-0.076

DDE

Average

Positive Composites

Number

Range

0.009

4
T-0.0.17

0.025

4
0.028-0.050

0.020

4
0.005-0.063

0.044

6
0.007-0.091

0.150

6

0.072-0.222

0.251

6

0.073-0.579

0.030

4
0.013-0.077

do.

TDE

Average

Positive Composites

Number

Range

0.006

2
0.005-0.031

0.020

3
0.031-0.050

0.014

3
0.010-0.050

0.012

5
0.004-0.029

1
0.053

0.018

2
0.051-0.058

0.018

2
0.008-0.065

do.

DIELDRIN

Average

Positive Composites

Number

Range

0.019

3
0.012-0.056

0.013

3
0.013-0.038

0.019

3
0.027-0.045

0.031

6

0.011-0.059

0.015

4
0.013-0.028

0.023

5
0.014-0.039

0.002

2
0.003-0.005

do.

HEPTACHLOR EPOXIDE
Average
Positive Composites

Number

Range

0.004

3
0.004-0.014

0.008

3
0.010-0.019

0.010

3

0.009-0.028

0.019

6

0.012-0.029

0.003

2
0.008-0.009

1
0.022

0.031

3
0.014-0.077

do.

BHC

Average

Positive Composites

Number

Range

0

1
0.032

0.019

4
0.008-0.072

0.026

5

0.016-0.057

0

1
0.026

0

do.

TOTAL BROMIDES

Average

Positive Composites

Number

Range

15.8

5
3.5-31.7

2.8

5
0.5-8.6

9.4

6

4.9-13.8

4.8

6

1.5-8.2

3.7
6

1.1-9.3

3.3
6

1.0-9.8

4.8
4

1.0-21.4

1.9

4

0.5-5.4

Vol. 1, No. 2, September 1967

TABLE 2a. — Levels of Pesticide Residues Commonly Found— In Food Class. Region, and Saniplinf; Period — Continued

PESnCIDE

Boston
1965 1966

Kansas City
1965 1966

Los Angeles
1965 1966

Baltimore
1966

Minneapolis
1966

II. MEAT. FISH AND POULTRY (17-23% fat) ■
Residues in Paris Per Million — Fat Basis

DDT

Average

Positive Composites

Number

Range

0.247

6
0.010-0.862

0.679

6
0.503-1.180

0.096

6

0.068-0.182

0.313

6
0.056-0.874

0,228

4
0.306-0.406

0.4.37

6
0.128-1.390

0.096

3
0.068-0.165

1
0.476

DDE

Average

Positive Composites

Number
Range

0.113

6
0.003-0.328

0.317

5
0.230-0.726

0.095

6
0.041-0.156

0.252

6
0.028-0.868

0.437

6

0.020-0.915

0.513

6

0.231-0.997

0.04S

4
0.017 3.066

1
0.S80

TDE

Average

Positive Composites

Number

Range

0.098

4
0.055-0.251

0.446

5
0.36I-0.78I

0.057

5
0.031-0.115

0.092

6

0.018-0.246

0.109

4
0.118-0.204

0.158

5
0.047-0.697

0.016

2
0.030-0.034

1
0.533

DIELDRIN

Average

Positive Composites

Number

Range

0.046

6

0.003-0.142

0.116

5
0.097-0.203

0.013

3
0.012-0.036

0.039

5
0.024-0.070

0.068

4
0.006-0.141

0.030

5
0.028-0.053

1
0.008

0

HEPTACHLOR EPOXIDE

Average

Positive Composites

Number

Range

0.015

5
0.002-0.059

0.097

5
0.053-0.290

0.012

3
0.011-0.049

0.021

6

0.010-0.030

0.031

4
0.018-0.082

0.017

4
0.006-0.051

1
0.006

0

BHC

Average

Positive Composites

Number
Range

0

0.140

6
0.070-0.203

0.047

5
0.027-0.141

0.022

3

T-0.063

I
0.065

1
0.043

0

0

LINDANE
Average
Positive Composites

Number
R:mgc

O.OOI

3
0.001-0.002

0.024

2
0.080-0.061

0

0

1
0.111

1
0.027

0

0

TOTAL BROMIDES

Average

Positive Composites

Number

Range

9.0

4
T-23.4

4.0

6
2.2-6.2

14.8

6

3.5-35.5

8.7

3
2.4-44.0

4.7

6
2.2-7.1

2.9

5
0.9-6.6

'8.7

4
0.5-34.3

'8.1

5
1.0-30.1

III. GRAIN AND CEREAL 2
Residues in Parts Per Million

DDT

Average

Positive Composites
Number

0.003

4

0.011

5

0.010
5

0.007
5

0.012

5

0.004
3

1

0

Range

T-O.OlO

0.006-0.024

0.005-0.021

T-0.015

0.008-0.026

0.004-0.012

0.007

DDE

Average

Positive Composites
Number

0

<0.001

2

0.002

3

0.002
4

<0.(X)l

->

O.OOI

->

0.002

2

0

Range

T-0.002

T-0.007

T-0.004

T-0.002

0.001-0.002

0.002-0.004

TDE

Average

Positive Composites
Number

0

1

0.004
3

0.001
3

1

1

0

Range

T

T-0.013

T-0.005

0.003

0.002

0.002

DIELDRIN

Average

Positive Composites
Number

1

1

0.005
5

1

0.001
2

1

0

Range

0.007

0.003

T

T-0.018

0.004

0.002-0.004

0.(X)6

Pesticides Monitoring Journal

TABLE 2a. — Levels of Pesticide Residues Commonly Found — iv Food Class, Region, and Sanipliiin Period — Continued

Pesticide

Boston
1965 1966

Kansas City
1965 1966

Los Anoeles
1965 1966

Baltimore
1966

Minneapolis
1966

III. GRAIN AND CEREAL :^( Continued)
Residues in Parts Per Million

LINDANE
Average
Positive Composites

Number

Range

0.004

4
T-0.012

0.007

5
0.004-0.014

0.009

6
T-O.0I6

0.018

6

0.005-0.028

0.007

4
0.001-0.032

O.OOI

3
0.002-0.003

0.004

3
0.002-0.009

0.006

3
0.006-0.010

MALATHION
Average
Positive Composites

Number

Range

0

1
0.035

0

0.018

4

0.013-0.035

0

0.041

4
0.025-0.153

0

0.011

2

0.024-0.018

TOTAL BROMIDES

Average

Positive Composites

Number

Range

20.7

6

10.0-31.2

15.7

6

11.2-20.4

52.0

6
10.4-111.0

12.2

6
1.1-18.6

6.5

6
4.4-9.6

6.8

6
2.4-11.7

•17.4

4
4.0-51.4

»19.5

5

2.3-66.7

IV. POTATOES =

Residues in Paris Per Million

DDT

Average

Positive Composites

Number

Range

0

0.003

4
T-O.OlO

0

0

1

0.006

1
0.007

1
0.004

0

DDE

Average

Positive Composites

Number

Range

0

1
T

0

1
0.010

0.001

4
T-0.002

1
0.003

1
0.005

0

DIELDRIN

Average

Positive Composites

Number

Range

0

1
0.003

1
0.006

<0.001

3
T-0.002

1
0.003

0.001

3
T-0.002

0

0

TOTAL BROMIDES

Average

Positive Composites

Number

Range

9.0

5
7.4-28.0

4.1

5
1.1-13.1

18.4

6

1.5-38.0

7.7

4
1.1-41.0

5.6

4
4.0-17.6

1.8

3
0.7-5.7

S9.7

3
4.0-38.3

'15.2

5
1.3-68.5

V. LEAFY VEGETABLES =
Residues in Parts Per Million

DDT

Average

Positive Composites

Number

Range

1
0.008

0.017

2
0.009-0.099

0.004

2
0.004-0.017

0.004

2
0.006-0.016

0.017

4
0.010-0.047

0.022

5
0.009-0.048

0.007

2
0.006-0.022

0.016

2
0.028-0.035

DDE

Average

Positive Composites

Number

Range

0.007

3
0.019-0.025

0.004

2
0.002-0.023

0.007

3
0.003-0.032

0.007

2
0.006-0.033

0.002

3
0.003-0.006

0.005

5
0.004-0.008

0.002

2
0.002-0.006

1
0.015

TDE

Average

Positive Composites

Number

Range

1
T

0

0.061

3
0.006-0.291

0.005

3
0.001-0.024

1
T

0.004

2
0.008-0.014

0.001

2
0.002-0.003

0.011

3
0.012-0.017

TOTAL BROMIDES

Aveiage

Positive Composites

Number

Range

4.6

5
1.3-16.3

3.3

6

1.3-7.2

4.4

6

1.7-10.5

2.0

6

0.7-3.4

1.9

5
0.7-6.8

1.3

5
0.6-2.9

3

1

14.8

»1.7

4
0.7-5.0

Vol. 1, No. 2, September 1967

TABLE 2a. Levels of Pesticide Residues Commonly Foiiml — hy Food Class. Region, and Samplinf; /'cr/of/— Continued

Pesticide

Boston
1965 1966

Kansas City
1965 1966

Los Angeles
1965 1966

Baltimore
1966

Minneapolis
1966

VI. LEGUME VEGETABLES =
Residues in Paris Per Million

DDT

Average

Positi\e Composites

Number

Ranxe

1
(I.IBSI

1
0.004

0

0

0 024

4
T-0.126

I
0.010

1
".i49

0

IDE

Average

Positive Composites

Number

Range

1

n.(>5l

1

0.010

0

1
T

1
11.006

0.001

3
0.001-0.004

1

o.oe-'

0

TOTAL BROMIDES
Average

Positive Composites
Number
Range

4.4

4
2.1-14.1

3.4

5
1.6-12.0

6.1

6
1.0-17.9

1.4

6
0.5-2.4

4
0.9-15.2

0.7

4
0.7-1.7

••■2.3

3
0.5-9.9

»4.6

4
1.1-14.5

VII. ROOT VEGETABLES =
Residues in Parts Per Million

DDT
Average
Positive Composites

Number
Range

1
0.009

0

0.018

2
0.039-0.071

0.003

2
0.007-0.011

0.002
0.003-0.009

0.005

2
0.006-0.021

0

0

DDE

Average

Positi\ e Composites

Number

Range

1

0.010

0

0.009

2
0.0204)051

0.003

4
T-O.OU

0,010

4
11,004-0.045

0.003

4
T-0.009

0

0

DIELDRIN

Average

Positive Composites

Number

Range

1

O.IX)8

0

1
0.018

0.008

4
0.003-0.028

1
0.004

0.002

3
0.001-0,005

0

0

TOTAL BROMIDES
Average

Positit e Composites
Number
Range

5,6

5
4.3-9.3

4.1

4
5.5-7.2

10.9

6

2.8-22.1

3.1

5
2.1-5.4

1.5

3
2.6-3.5

2.8

5
1.5-7.5

' 3.0

3
0.7-8.9

"4.1

4
0.6-17.0

VIII. GARDEN FRUITS =
Residues in Parts Per Million

DDT

Average

Positive Composites

Number

Range

1

0.011

3
0.008-0.033

0.014

4
T-0.034

0.034

4
0.018-0.149

0.059

4
0.011-0.168

0.032

5
0.018-0.086

0.048

6

0.022-0.115

0.013

2
0.012-0.038

0.007

2
0,014-0.030

DDE
Average
Positive Composites

Number

Range

1

r

0.001
2

T-i>"no2

1
0.002

0.0 1 1

4

T-0.064

0.003

6
■I'.0.(X)9

0.004

3
T-0,023

0.002

2
0.003-0.005

0

roil

Average

Po\ili\e Composites

Number

Range

0.017

3
0.017-0.048

0,006

3
0.005-0.020

0,010
0.009-0.049

0.072

6
T-0.338

0.001

3
T-0.003

0.006

-»
0.011-0.023

1
0.015

0.005

2
0.01.3^.017

DIELDRIN

Average

Positive Composites

Number

Range

0.002

2
T-0.012

1
0.004

0.004
2

0.010-0.011

0.003

3
0.002-0.013

0,002

4
T-0.(K)5

<0.001
0.001-0.003

t
0,004

0.006

2
0.006-0.017

LINDANE

Average

Positive Compt}\iles

Number

Range

O.IK)3
O.U08-O.OII

0.001

3
T-O.005

0.007

3

0.008-0.025

0.003

4
T-0.012

0.003
0.002-0.017

<0.001

2
T-O.OOl

1
0.002

0,002
0.004-0.005

Pesticides Monitoring Journal

TABLE 2a. — Levels of Pesticide Residues Commonly Found — by Food Class, Region, and Sampling Period — Continued

Boston
1965 1966

Kansas City
1965 1966

Los Angeles
1965 1966

Baltimore
1966

Minneapolis
1966

VIII. GARDEN FRUITS =— (Continued)
Residues in Parts Per Million

TOTAL BROMIDES

Average

9.1

3.0

6.4

3.5

1.5

2.5

»2.6

«2.3

Positive Composites

Number

5

5

6

6

4

5

3

5

Range

5.7-18.9

1.1-7.5

4.0-8.3

1.2-7.5

1.7-2.8

1.7-7.1

1.5-9.2

0.5-7.2

IX. FRUITS =
Residues in Parts Per Million

DDT

Average

Positive Composites

Number

Range

0.008

3
0.008-0.027

0.012

4
0.007-0.035

1

0.006

1
T

0.008

5
0.007-0.019

0.010

2
0.012-0.045

0.006

3
0.004-0.014

0

DDE

Average

Positive Composites

Number

Range

<0.001

2
T-0.003

0.001

2
0.002-0.004

1

0.005

0.001

4
T-0.005

0.001

5
T-0.003

0.008

3
0.003-0.043

0.003

3
0.001-0.006

0

TDE
Average
Positive Composites

Number

Range

0

0

0

1

0.007

1
0.004

1

0.007

0.003

3
0.002-0.006

0

ALDRIN
Average
Positive Composites

Number

Range

0.008

4
0.003-0.020

0.015

3
0.007-0.070

1
0.007

0

0.001

3
0.002

1
0.002

0.004

2
0.005-0.012

0

KELTHANE
Average
Positive Composites

Number

Range

0

0.032

3
0.013-0.107

1
0.166

0.068

6

0.016-0.161

0

0.041

2
0.032-0.212

1
0.015

0

TOTAL BROMIDES
Average

Positive Composites
Number
Range

3.1

2
3.2-15.3

3.2

5
0.8-10.2

7.9

6
1.2-31.4

3.2

6

0.7-6.4

1.0

4
0.7-2.4

0.6

3
0.7-1.9

3
1

20.4

•6.0

4
1.1-25.2

X. OILS, FATS AND SHORTENING =
Residues in Parts Per Million

DDT

Average

Positive Composites

Number

Range

1
0.028

0.010

3
T-0.032

0.008

2
0.008-0.038

0.008

5
T-0.018

1
0.049

0.011

2
0.027-0.038

0.010

2
0.009-0.031

0

DDE
Average
Positive Composites

Number
Range

1

0.014

0.003

2
0.009-0.010

0.003

2
0.004-0.012

0.004

6
T-0.011

1
0.006

0.011

3
0.013-0.029

0.006

3
0.007-0.009

0

TDE

Average

Positive Composites

Number

Range

1
0.018

0.025

2
0.034-0.117

0.010

2
0.025-0.037

0.007

5
T-0.027

1
0.032

0.021

3
0.025-0.068

1
0.010

0

HEPTACHLOR EPOXIDE

Average

Positive Composites

Number

Range

1

0.004

0

0.001

3
0.001-0.004

0.001

3
T-0.004

1
0.002

0

0

0

TOTAL BROMIDES

Average

Positive Composites

Number

Range

12.6

6
1.1-29.0

5.3

5
1.9-11.8

53.0

5
7.2-261.0

7.5

6
0.9-15.2

4.4

5
1.7-9.8

6.2

2
5.7-7.3

3 12.8

3
1.3-48.8

»23.0

5
1.1-90.8

Vol. 1, No. 2, September 1967

TABLE 2a. Levels of Pesticide Residues Commonly Found — by Food Class, Region, and Sampling Period — Ci)ntinued

PESnCIDE

Boston
1965 1966

Kansas City
1965 1966

Los ANOEtES

1965 1966

Baltimore
1966

Minneapolis
1966

XI. SUGARS AND ADJUNCTS -
Residues in Parts Per Million

2, 4.D
Average
Positive Composites

Number

Ranee

0

1
0.01

0.020

4

0.020-0.040

0.030

4
0.020-0.058

0.07

4
0.04-0.16

0.038

3
0.057-0.100

0

0

TOTAL BROMIDES

Averace

Positive Composites

Number

Range

12.8

6
4.0-26.4

11.2

6

7.0-17.7

30.8

6
12.0-55.1

10.5

6
6.0-16.5

4.3

6
0.7-9.2

3.8

5
2.3-7.7

M2.8

3
2.1-55.4

"29.7

5
0.7-117.0

XII. BEVERAGES 2
Residues in Parts Per Million

TOTAL BROMIDES

Avcrape

5.7

4.1

7.9

1.5

s

»0.5

Positive Composites

Number

3

4

5

1

2 0

1

3

Range

1.2-16.2

1.3-13.7

2.8-15.0

3.2

0.9-8.2

8.7

0.5-1.2

^ Six composilc samples examined each year at Boston, Kansas City, and Los Angeles: four composite samples examined October 1965-April
1966 at Baltimore; for bromides, five composite samples examined beginning August 1965-April 1966 at Baltimore and Minneapolis.

- Six composite samples examined each year at Boston. Kansas City, and Los Angeles; four composite samples examined October 1965-April
1966 al Baltimore and Minneapolis.

^ Five composites examined beginning August 1965.

Note: 1965 = June 1964-April 1965
1966 = June 1965-April 1966

TABLE 2b. — Pesticides Found Infrequently —

by Food Class, Region, and Sa

mpling Period

No.

No.

Pesticide

District

Com-
posites

Year

Amount (ppm)

Pesticide

District

Com-
posites

Year

Amount (ppm)

I (a). DAIRY PRODUCTS (8-13% fat) '

III (a). GRAIN AND CEREAL i

Residues in Parts Per Million — Fat Basis

Residues in Parts Per Million

ALDRIN

Kansas City

1

1966

T

ALDRIN

Boston

1965

0.001

LINDANE

Kansas City
Boston

2

1

1965-1966
1965

T, 0.210
0.006

Baltimore
Los Angeles

1966
1966

0.016
0.014

MCP

Boston

1

1966

0.583

BHC

Kansas City

1966

T

Kansas City

1

1966

0.039

CARBARYL

Kansas City

3

1965

0.42, 0.20, 0.30

METHOXY=

Kansas City

3

1966

T, T 0 073

DIAZINON

Kansas City

1966

0.024. 0.024

CHLOR

Minneapolis

1966

0.004, 0.030

PC?

I

1966

0.310

DITHIOCAR=

Kansas City

1965

0.5

Kansas City

1

1966

0.009

BAMATES

2,4-DB

Los Angeles

1

1966

0.025

ENDRIN

Boston

1966

0.001

2,4. 5-TP

Boston

2

1966

0.018, 0.029

HEPTACHLOR

Los Angeles
Kansas City

1966
1965

0.004

II (a). MEAT, FISH AND POULTRY (17-23% fat) '

Los Angeles

1965

T

Residues in Parts Per Million — Fat Basis

HEPTACHLOR

Boston

1966

0.005

EPOXIDE
MCP

Kansas City
Boston

1966
1965

T, 0.005

ALDRIN

Kansas City

I

1965

0.008

0.10

DIAZINON

Kansas City

1

1966

0.051

METHOXY=

Boston

1966

O.0O4

ENDRIN

Kansas City

1

1965

T

CHLOR

Kansas City

1966

0.007

HF.PTACHLOR

Kansas City

1

1965

0.008

PCNB

Boston

1966

0.005

PCP

Kansas City

2

1965

0.01,0.03

PCP

Kansas City

1965-1966

0.02, 0.036

Boston

2

1966

0.005,0.051

Boston

1966

0.004

Los Angeles

1

1966

0.051

PERTHANE

Kansas City

1966

0.057, 0.049

RONNEL

Kansas Citv

1

1966

0.011

2,4-DB

Kansas City

1966

0.013

TETRADIFON Los Angeles

1

1966

0.076

ARSENIC Los Angeles

6

1965-1966

0.12,0.2,0.1,

RONNEL

Kansas City

1966

T

(AiiOi)

0.1,0.1,0.2

ARSENIC

Boston

1965

0.10

1 Kansas City

'

1966

0.5

(AsjOs)

Los Angeles

1966

0.10

10

Pesticides Monitoring Journal

TABLE 2b. — Pesticides Found Infrequently — by Food Class, Region, and Sampling Period — Continued

No.

No.

Pesticide

District

Com-
posites

Year

Amount (ppm)

Pesticide

District

Com-
posites

Year

Amount (ppm)

IV (a). POTATOES!

VI (a). LEGUME VEGETABLES'

Residues in Parts Per Million

Residues in Parts Per Million

BHC

Kansas City

1

1966

0.008

ALDRIN

Boston

1966

0.006

CARBARYL

Kansas City

I

1965

0.28

DDE

Los Angeles

1965-1966

0.003, T. 0.002,
0.002

CIPC

Kansas City

2

1966

0.360,0.199

Baltimore
Boston

1966
1966

0.003
T, 0.003

ENDRIN

Kansas City
Los Angeles

1
3

1965
1965-1966

T

0.005, 0.002,

Kansas City

1966

T

0.006

DIELDRIN

Los Angeles

1965

0.002

Boston

1

1966

0.004

Kansas City

1966

T

HEPTACHLOR

Kansas City

4

1965-1966

0.015, 0.020,

HEPTACHLOR

Los Angeles

1966

0.001

EPOXIDE

0.002, T

EPOXIDE

LINDANE

Boston
Baltimore

1

1965
1966

0.008
0.011

LINDANE

Boston

1966

T

Kansas City

2

1966

T, 0.002

ARSENIC

Boston

1965

0.11

Los Angeles
Los Angeles

1
1

1966
1966

T

0.003

(AsiiOs)

PARATHION

TCNB

Boston

1

1965

0.216

VII (a). ROOT VEGETABLES'

Baltimore

1

1966

0.370

Residues in Parts Per Million

TDE

Los Angeles

Los Angeles

2

1

1965-1966
1966

T. 0.001
4.7

ARSENIC

(AsiOa)

CARBARYL
CHLOR=

Kansas City
Kansas City

2
1

1965-1966
1966

0.20,0.10

0.010

BENSIDE

V (a). LEAFY VEGETABLES >

ENDRIN

Kansas City

2

1965-1966

T, 0.052

Residues in Parts Per Million

MALATHION

Kansas City

1

1966

0.022

TCNB

Kansas City

1

1965

0.011

BHC

Kansas City

1

1965

0.015

Los Angeles

1

1966

T

CARBARYL

Kansas City

2

1965

0.3, 0.2

TDE

Kansas City
Los Angeles

2
1

1965-1966
1965

0.021,0.009
0.004

CHLOR=
BENSIDE

Kansas City
Los Angeles

Los Angeles

2

1965
1965

1966

0.023, 0.038
0.002

0.006

ARSENIC

(As^Oa)

Boston

Minneapolis

1

1

1965
1966

0.10
0.10

DACTHAL

DIAZINON

Los Angeles

1966

0.015, 0.012

Minneapolis

1966

0.031

VIII (a). GARDEN FRUITS'

DIELDRIN

Baltimore
Kansas City

Kansas City

1966
1966

1965

0.002
T

0.4. 0.7, 0.8

Residues in Parts Per Million

DITHIOCAR=
BAMATES

ALDRIN

Boston

1966

0.005

2,4-D

Kansas City
Boston

1965
1966

T
0.017

ARSENIC

(As:Os)

Minneapolis

1966

0.10

ENDRIN

Kansas City

1965

T

BHC

Kansas City

1965

0.004

HEPTACHLOR

Kansas City

1965

0.004

CARBARYL

Kansas City

1965

0.19

EPOXIDE

CHLORDANE

Boston

1965-1966

0.033, 0.006

LINDANE

Los Angeles
Boston

1965
1966

0.004
T, 0.005

Los Angeles

1966

0.002

Minneapolis

1966

0.012

DIAZINON

Minneapolis

1966

0.005

MALATHION

Kansas City

1966

0.017

ENDRIN

Kansas City

1965-1966

0.007, 0.005

MCP

Boston

1966

0.114

HEPTACHLOR
EPOXIDE

Los Angeles
Boston

1965
1966

T
T

PARATHION

Boston

1966

0.012

Los Angeles

1966

0.016

ENDOSULFAN

Boston

1966

0.006, T

Minneapolis

1966

0.089

Los Angeles

1966

0.002

ENDOSULFAN

Los Angeles

1966

0.016

TOXAPHENE

Boston
Los Angeles

1966
1966

0.048
0.050

TOXAPHENE

Baltimore

1966

0.386

Vol. 1, No. 2, September 1967

11

TABLE 2b. — PesikUlcs Foiiml hijrequeiuly — Ay Food Class, Region, and Sampling Period — Continued

Pesticide

1

DisrmcT

No.
Com-
posites

Year

Amount (ppm)

Pesticide

DlSTKICT

No.
Com-
posites

Year

Amount (ppm)

IX (a)

. FRUITS >

X (a). OILS. FATS AND SHORTENING ' -(Continued)

Residues in

Parts Per Million

Residues in Parts Per

Million

CARBARYL

K.ii:!..is City

4

1965-1966

0.19, 0.20, 0.10,

0.20
0.17
0.05

0.004

0.004. T. 0.002
T, T, T

0.019

PERTHANE

Kansas City

1

1966

0.032

DACTHAL

Boston

Los Angeles

Boston

Boston
Kansas City

Boston

I

1

1

3

3

1

1966
1966

1966

1965-1966
1966

1966

2,4-D
TBA

Boston
Kansas City

1
1

1965
1965

0.030
0.02

DIELDRIN

ETHION

XI (a). SUGARS AND ADJUNCTS'
Residues in Parts Per Million

LINDANE

Kansas City
1 OS Angeles
Boston

I
2
2

1965
1966
1966

0.009

0.002. 0.005
T, 0.002

ALDRIN
BHC

Kansas City
Kansas City

1
I

1965
1965

0.003
0.015

PC SB

Kansas City

1

1965

0.00.1

CARBARYL

Kansas City

4

1965-1966

0.7, 0.2, 0.1, 0.2

PERTHANE
TETRADIFON

Boston

Boston
Los Angeles

2

1
2

1965-1966

1965
1965-1966

0.016. 0.007

0.044
0.006.0.011

DDT
DDE

Kansas City
Los Angeles

Los Angeles

2
3

3

1965
1965-1966

1965-1966

0.021,0.085
T, 0.005, 0.008

0.003, T, 0.004

ENDOSULFAN

Kansas City

1

1966

0.014

DIELDRIN

Los Angeles

2

1965-1966

T, 0.002

ARSENIC
(As:03)

Boston

1

1965

0.18

HEPTACHLOR
EPOXIDE

LINDANE

Los Angeles
Kansas City

Kansas City
Los Angeles
Boston

2
1

2
1

1

1965
1966

1965-1966
1965
1966

0.002, T
T

X (:

1). OILS, FAT
Residues in

S AND S
Parts Per

HORTENl

Million

NG'

T, T

0.001
T

MCP
RONNEL

Boston
Los Angeles

1

1

1966
1966

0.022

ALDRIN

Kansas City

2

1965-1966

T, T

T

BHC

Kansas City

1

1965

0.007

TDE

Los Angeles

2

1965

0.012, T

DIELDRIN

Kansas City
Los Angeles

Kansas City
Los Angeles

Kansas City

Kansas City
Los Angeles

Boston
Kansas City
Minneapolis

3
4

2
1

1

1

1

2

1

1965-1966
1965-1966

1965-1966
1966

1965

1965
1965

1966
1966
1966

T, 0.005, T
0.007, 0.070,
0.005, 0.009

0.017, 0.006
0.012

0.002

0.003
0.004

0.053

0.053, 0.013
0.18

ARSENIC

(AS.O3)

Boston

1

1966

0.1

ENDRIN

XII (a). BEVERAGES'
Residues in Parts Per Million

HEPTACHLOR
LINDANE

MALATHION

CARBARYL

DDE

HEPTACHLOR
EPOXIDE

Kansas City

Los Angeles
Kansas City

4

1

1

1965-1966

1965
1966

0.37, 0.50, 0.50,
0.40

T

T

PCP

Boston
Los Angeles

I
1

1966
1966

0.012
0.193

LINDANE
PCP

Kansas City
Boston

1
I

1966
1966

T
0.02

' Six composite s;:mples examined each year at Boston, Kansas City,

1966 at Baltimore and Minneapolis.
Note: 1965 = June 1964-April 1965
1966 = June 1965-April 1966

and Los Aiitieles; four composite samples examined October 1965-April

A cknowledgment

The authors gratefully acknowledge the analytical work
from the FDA Laboratories in Baltimore, Md.; Boston,
Mass.; Kansas City, Mo.; Los Angeles, Calif.; and
Minneapolis, Minn.

LITERATURE CITED

(1) niiRgan. R. £., //. C. Barry, and L. Y. Johnson. 1966.
Pcslicidc residues in total diet samples. Science 151:101.

(2) Diif-aan. R. E.. and F. J. McFurland. 1967. Assessments
include raw food and feed commodities, market basket
items prepared for consumption, meat samples taken
at slaughter. Pesticides Monit. J. 1( I ):!.

13) CidPrida. Laura. 1964. A flame ionization detector
highly sclcclive and sensitive to phosphorus— a sodium

thermionic detector. J. Ass. Offic. Agr. Chem. 47:293;
//)/(/. ]964. Investigation of two gas chromatographic
techniques for the determination of organophosphate
pesticide residues. 47: 1 1 1 2.

14) Mills, P. A., J. H. Onley, and R. A. Gailhcr. 1963.
Rapid method for chlorinated pesticide residues in
non-fatty foods. J. Ass. Offic. Agr. Chem. 46:186-191.

15) Kovacs. Mariin F., Jr. 1964. Thin layer chromatography
for organo thiophosphate pesticide residue determina-
tion. J. Ass. Offic. Agr. Chem. 47: 1097.

(6) Finoccliiuro, J. M.. and \V. R. Benson. 1965. Thin layer
chromatographic determination of carbaryl (Scvin) in
some foods. .1. Ass. Offic. Agr. Chem. 48:736.

(7) Diingan, R. £.. and J. R. IVealhcrwa.x. 1967. Dietary
intake of pesticide chemicals. Science (Accepted for
publication).

12

Pesticides Monitoring Journal

Chlorinated Hydrocarbon Pesticide Residues

in or on Alfalfa Grown in Soil With a Previous History

of Aldrin and Heptachlor Application

R. J. Moubry", G. R. Myrdal', and H. P. Jensen=

ABSTRACT

Samples of soil, alfalfa, and alfalfa roots were collected from
acreage with a past history of aldrin and heptachlor applica-
tion. The samples were analyzed with the final determination
by gas liquid chromatography (GLC). Data obtained are
presented on both the wet, or as is, and the dry weight basis.

LOW level dieldrin residues in milk from herds located
in corn producing areas of the State prompted an in-
vestigation into the possible contamination of alfalfa
grown in soil with a past history of aldrin use. The
common use of aldrin involved the placement of in-
secticidal granules in the corn row as a narrow band
behind the seed shoe. In an effort to determine the effect
of this practice, personnel of the Wisconsin Department
of Agriculture, in cooperation with J. W. Apple, Pro-
fessor of Entomology. College of Agriculture, Univer-
sity of Wisconsin, collected samples from an alfalfa
field which had a known soil insecticide application and
cropping history.

A 40-acre field in Columbia County was selected for
this study. Corn had been grown on this field in 1962,
1963, and 1964. Pesticide application was by band
treatment, with 1 lb/ acre heptachlor in 1962, 1 lb/ acre
aldrin in 1963, and 1 lb/ acre aldrin in 1964. In 1965
the field was seeded with alfalfa, with oats planted as a
nurse crop.

The east one-half of the field was sampled in a diagonal
pattern on August 30, 1965. Samples of soil (Carrington
silt loam), alfalfa and alfalfa roots were randomly col-
lected at approximately 20-foot intervals. Composites
of the samples of alfalfa and alfalfa roots were extracted

Wisconsin Department of Agriculture, General Laboratory Division,
4702 University Avenue, Madison. Wise. 53705.
Present address: Ciba Corporation, Agricultural Chemicals Test
Laboratories, Vero Beach. Fla. 32960.

and cleaned up by the acetonitrile extraction procedure
(1). Composites of the samples of soil were extracted by
the hexane-acetone procedure (2). The soil extracts were
cleaned up with Florisil il). Prior to analysis the alfalfa
and root samples were ground and mixed in a Hobart
food chopper. A portion of each homogeneous sample
was selected for a moisture determination. Analysis was
made on the wet weight basis. The dry weight residue
results were obtained by calculation, using the percent
moisture obtained for each sample.
The determination of the amount of pesticide residue
present in the sample was by GLC.

Conditions of Gas Liquid Chromatography Deter-
mination
Instrument — Jarrell-Ash, Model 28-710
Column — 4 ft. x 0.156 in. bore glass
Packing— 10% DC200 on 80-90 mesh Anakrom

ABS
Detector — Electron affinity, source 100 mc H^
Amplifier — Sensitivity 1 x 10"'' A, voltage, 18 v
Flow Rate — 196 ml Nn/min.; pressure, 30 lbs/

sq. in.
Temperatures — Injector, 240 C; oven, 203 C;
detector, 209 C

The presence of the pesticide residue in the samples was
confirmed by GLC, using different column systems.
These were (a) mixed bed consisting of one part 10%
DC200 on 80-90 mesh Anakrom ABS and two parts
5% QFl on 60-80 mesh Chromosorb W, AW; and (b)
5% QFl on 60-80 mesh Chromosorb W, AW.

The sample size used for GLC injection was selected to
provide detection and confirmation of residues at or
above 0.001 ppm on the wet weight or as is basis.

Vol. 1, No. 2, September 1967

13

TABLE 1. — Detection of cyclodiene insecticide residues in alfalfa and soil following use on corn

Residues in ppm

Sample

Wet Weight Basis

Dry Weight Basis

Hepta-

CHLOR

Hepta-

CHLOR

Epoxide

AlDRIN

DIEIDRIN

Hepta-
chlor

Hepta-

CHLOR

Epoxide

Aldrin

Dieldrin

Soil, Top Vy Layer

0.139

0.226

0.007

0.019

Soil, 6" Cores

0.211

0.221

0.032

0,036

Alfalfa Roots >— Group 1

0,098

0.460

0,014

0,142

0,293

\.?7n

0.041

0,424

Alfalfa Roots with Tops ' — Group 2

0.049

0.360

0.0 lU

(1,120

0.146

0.970

0.030

0.358

AUalfa '

0.020

0,IH).1

0,111

0.015

Alfalfa, Lower Half

0.031

0,004

0.165

0,020

Alfalfa. Upper Half '

0.010

0.001

0,059

0,008

Alfalfa «

0.010

0.002

0.061

0.012

* These roots were washed and separated into two groups. The aerial portions of the plants were removed from the first group; V2 to 1 inch

of the aerial portions were left remaining on the second group,
= This alfalfa was cut '/i inch above ground. It was thoroughly washed before analysis,
•■^ A portion of the alfalfa plants cut V2 inch above ground was randomly selected from the original sample (b). These plants ranged between

4 and 10 inches in height. Each plant was water washed and cut in half. The lower and upper halves were grouped, ground, and analyzed.
' This sample of alfalfa was collected at the same time and in the same manner as that defined in (b) except that it was cut Hi inches above

ground. This alfalfa was not washed prior to analysis.

Heptachlor l,4,5,6,7.8.8-heptachloro-3a,4,7,7a-tetrahydro^,7-methanoindene

Heptachlor epoxide 1 ,4,5.6,7.8.8-heptachloro-2,3-epoxy-3a,4,7,7a-tetrahydro^,7-methanoindan

Aldrin not less than 95% of I,2.3,4.10,l0-hexachloro-1.4.4a,5.8.8a-hexahydro-l.4-f«do-f-vo-dimethanonaphthalene

Dieldrin not less than 85^^ of l.:.,'<.4,10,I0-hexachloro-6.7-epoxy-l,4,4a,5,6,7.8.8a-octahydro-l,4-eiiifo-«o-5,8-dimcthanonaphthalene

Inasmuch as this was an exploraton' survey, recovery
studies were not conducted in conjunction with these
samples. The data presented are the results obtained
using the methodoiogy specified. The results are detailed
in Table 1.

In the analysis of forages in this laboratory we custom-
arily report results in the range of 0.005 ppm to 0.001
ppm as a "trace." In this study results below this level
are defined numerically, because they present informa-
tion useful in comparison to the amount of residue found
in the different portions of the plants. Data obtained
in this initial investigation showed aldrin residues in the
root and aerial portions of alfalfa grown in soil where
aldrin had been applied as a band treatment in corn
rows 1 to 2 years previously; similar treatment with
heptachlor 3 years previously also resulted in residues
in this crop.

Feeding of this forage to dairy animals will provide
a minor source of dieldrin to the diet which could be
a contributing cause of low level residue in the milk
production of these animals (3). These data also indi-
cated that the residue levels were higher in the roots

than in the aerial portion of the plant. Analysis of the
aerial portion of the plants divided into upper and
lower halves showed higher residues in the lower halves
of the plants. Washing of the aerial portions did not
shown any effect on the residue present. The data also
showed that residues in the soil were appreciably lower
in the top Vi-inch layer than in the upper 6 inches of
the soil.

LITERATURE CITED

(1) Barry. Helen C, Joyce G. Hundley, and Lorcn Y.
Johnson. Pesticide Analytical Manual Vol. 1, 2.21(B),
U. S. Depanment of Health, Education and Welfare,
FiK)d and Drug Administration, Washington. D.C.
20204.

(2) Lichlenstein, E. P., G. R. Myrdal, and K. R. Schultz.
1964. Effect of formulation and mode of application
of aldrin on the loss of aldrin and its epoxide from soils
and their translocation into carrots. J. Econ. Entomol.
57:133-136,

(.?) Williams, S., P. A. Mills, and R. t". McDowell. 1964.
Residues in milk of cows fed rations containing low
concentrations of five chlorinated hydrocarbon pesti-
cides. J. Ass. Offic. Agr. Chem. 47(6); 1124-1128.

14

Pesticides Monitoring Journal

PESTICIDES IN PEOPLE

Storage of DDT in The People of Israel'

M. Wassermann, Dora Wassermann, L. Zellermayer, and M. Gon

MEASUREMENT of the storage of chlorinated hydro-
carbon insecticides in the body fat constitutes a valuable
tool for the appraisal of exposure of the general popula-
tion to these compounds.

Their storage is encountered in populations of diffeient
continents all over the world. The main source of insec-
ticide absorption is the dietary intake, but air pollution
produced by the household use of insecticides may also
contribute to storage. Use of new analytical techniques,
especially gas chromatography, has revealed that, be-
sides DDT and its metabolite DDE, other organo-
chlorine insecticides are stored in the body fat of people
without known occupational exposure. The compounds
include: DDD and /3-isomer of BHC in the general
population of the USA (New Orleans) (14); BHC in
body fat in the USA (1.15.16) in France (13}, and India
(2); y-isomer of BHC in the general population of Eng-
land (23); dieldrin in body fat in the USA (1.15.16.24).
in Southern England (17,23) and in India (2); hepta-
chlor epoxide in persons in the USA (14.29). and in
India (2); and DDD and dieldrin — in some cases also
7-isomer of BHC — in the general population and in
farm workers of USA (Dade County, Florida) (8).
This paper reports on a further study that has been
carried out on the general population of Israel in order
to follow up the evaluation of organochlorine insecticide
storage in this country. A previous study was performed
by us on 254 specimens of fat tissue, obtained in 1963-
64, from persons without occupational exposure (26).
It revealed that, at that time in the body fat of the gen-
eral population of Israel, the mean concentration of

The systematic names of compounds mentioned in this paper are:

DDT l,l,l-trichloro-2,2-bis(p-chloroplienyl) ethane

DDD 1 .1 -dichloro-2.2-bis{ p-chlorophenyl ) ethane

DDE l,l-dichloro-2.2-bis(p-chlorophenyl) ethylene

BHC 1,2.3,4,5.6-hexachIorocyclohexane. mixed isomers

Dieldrin not less than 85% of l,2,3.4.IO,IO-hexa=

chloro-6.7-epoxy-l,4,4a,5,6,7.8.8a-octa=

hydro- 1 ,4-eHdo-e.vo-5,8-dimethanonaphthalene
Heplachlor epoxide l.4,5,6.7,8,8-hepta=

chloro-2,3-epoxy-.3a,4,7,7a-tetrahydro-4,7-

methanoindan

DDT was 8.5 ppm, that of DDE was 10.7 ppm and
that of DDT-equivalent (expressed as the numerical
sum of DDT and DDE) was 19.2 ppm; DDE averaged
55.6% of the total DT-derived material.

Material and Metlwds

The survey was carried out on 204 samples (144
autopsy and 60 biopsy specimens) from five hospitals
and the Forensic Medicine Institute. They were ob-
tained in 1965 and 1966 from persons without known
occupational exposure to pesticides. A survey sheet
containing information regarding name, sex, country of
origin, occupation, dietary habits, and operative diag-
nosis or cause of death was completed for each sample.
Each specimen was preserved in 4% formaldehyde
(Hayes et al. (12) have shown that this method of pres-
ervation is suitable for such survey purposes). The
analyses for DDT were performed by the Schechter-
Haller spectrophotometric method with the modification
described by the Technical Development Laboratories,
Communicable Disease Center (25). A Shimadzu XV
type automatic Recording Spectrophotometer model
SV.50A served to record the visible spectrum. Since the
method fails to distinguish between DDE and DDD, the
values given for DDE must be considered to represent
the sum of the two compounds.

The distribution of samples according to age and geo-
graphic origin is summarized in Table 1. From the

TABLE \. — Distribution of samples according to age and
geographic origin

From the Department of Occupational Health, the Hebrew Univer-
sity, Hadassah Medical School, Jerusalem, Israel.

Area of Origin

Ace Group

No. OF

(IN YEARS)

Samples

Israel

Europe

Asia

Africa

0- 9

71

69

1

1

10- 19

■}

1

—

—

1

20 ■ 29

5

1

1

1

2

30-39

11

—

3

5

3

40-49

24

1

13

6

4

50-59

31

4

23

2

2

60-69

37

4

15

9

9

70 - 79

18

2

8

5

3

80-89

5

—

5

—

—

TOTAL

204

82

68

29

25

Vol. 1, No. 2, September 1967

15

epidemiological point of view, it wus important to deter-
mine thai these samples were from persons who them-
selves, or whose parents, immigrated at least 8 years
previously, and it can be assumed they have had un-
changed living conditions since immigration. There are
great ditferences in the dietary and cooking habits
among the dilferent ethnic groups of this country. How-
ever, the basic foods for the entire population of the
country have a similar origin, and it appears therefore
that these people have similar exposure to the dietary
intake of DDT.

In the previous study we found no significant differences
between the mean values of DDT-derived material in
different ethnic groups. In this study, likewise, there
were no differences by ethnic group. For this reason
the data in this paper are presented by sex and age only.
For purposes of comparison with the previous study in
which there were only three cases in the age group 0 - 9
years, mean values are given separately for the groups
0 - 9 years and 10-89 years.

The group 0- 9 years, consisting of 71 cases, or 34.4%
of the total number of cases, provided a valuable op-
portunity for studying storage at these ages (Table 2).

Results

A total of 204 samples of fat tissue originating from
persons without known occupational exposure have been
analyzed for DDT-derived material. The results are
summarized in Table 2.

There are no significant differences between the mean
values found in different ethnic groups.

In the general population of Israel, age 10-89 years,
the mean of total DDT-derived material in 133 samples
is 18 +- 12.6 ppm. DDT is present in an average con-
centration of 8.2 zt 6.1 ppm, and DDE in 9.9 ± 7.1
ppm. DDE averages 53.9 ± 6.8% of the total DDT-
derived material.

There is no significant difference between the present
results and those obtained in our previous study on
Israelis (p>0.1 ). In the period 1965-66. the storage of
DDT-derived material continued to be maintained at a
high level in comparison with those levels reported
during the last decade in other countries with the
exception of India (Table 3).

In the population aged 10- 89 years, there is no signifi-
cant difference at the 5% level in the DDT-derived
material stored in difference age groups.

Males store significantly higher amounts of DDT-derived
material than females, namely, for DDT, p<0.02-
DDE, p<0.02: DDT j DDE, p = 0.01. For percent
of DDE the difference was not significant (p>0.05).

When the age group considered is restricted to 60 - 69

yearr. there is a significant difference (p<0.05) for
DDE and for DDT -\- DDE. There is no significant
difference lor DDT and for the DDE percent (p>0.05).

In the age group 0-9 years, the average concentration
of DDT -f DDE is 10.2 ± 9.2 ppm. The mean for
DDT is 4.6 ± 4.2 ppm and for DDE is 5.6 ± 5.9
ppm. The DDE percent is 53.3 ± 7.9.

As far as concentrations of DDT. DDE. and DDT -(-
DDE are concerned, a significant difference (p<0.001)
does exist in the storage of DDT-derived material be-
tween the group aged 0-9 years and that of 10-89
years even though certain unusually high values found
in the 0-9 age group are used in the calculations. From
the total of 71 cases belonging to this age group, 69
were aged from 0-2 years (Table 2). No significant
difference was found in the storage of DDT-derived
material among stillborns, neonates, and infants.

Mathematically there is a significant difference
(p<0,05) for the DDE percent in the female sex
between the stillborn, neonate, and infant categories.
In stillborns. there is a significant difference (p<0.05)
between sexes for DDE percent.

However, the number of cases in every category is too
small to justify a biological conclusion.

Comments

The data presented in Table 3 suggest that DDT is a
current constituent of human fat in the general popula-
tion of the world at this time.

Twelve samples of this study were from stillborns and
24 from neonates (15 in the first week of life). DDT-
material was found in all these samples and. in fact, in
all the samples analyzed in this study. Denes (3) found
DDT in a day-old neonate and Halacka el at. (9) in
three neonates. In our previous study in 1963-64, we
found 16.1 ppm DDT-derived material in the fat tissue
of a day-old neonate.

A note on the results of 50 samples as part of the present
study has been presented at the First World Congress on
Air Pollution (27). At that lime we found a mean of
10.2 ppm DDT-derived material in the fat tissue of still-
borns, neonates, and infants of Israel.

Fiserova-Bergerova et al. (S) found in the fat tissue of
four stillborns and two fetuses 5.65 ppm total DDT.
Zavon el al. (30) in 64 samples originating from children
dying between the 36th week of gestation and 2 weeks
postpartum, have found DDT-derived material,
heptachlor epoxide, and dieldrin. All these studies show
that the stored DDT-derived material in fetuses, new-
borns, and infants in their first weeks are in a lower
concentration than in the adult population. In animal
studies Finnegan et iil. (7) found DDT in the offspring
of dogs and Pillmorc cl til. (20) in rabbits.

16

Pesticides Monitoring Journal

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r-, vo

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Range

Mean-S
N

Range
Mean-S

N

Range
Mean-S

N
Range

Mean-S
N

Range
Mean-S

N

Range

Mean-S
N

Range
Mean-S

N

2

cfl

Cfl

2

a

a

n

4>

>.

1)

>.

>^

>,

>%

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r-

00

Vol. 1, No. 2, September 1967

17

TABLE 3. Conceniralion of DDT and its metabolite DDE in the body jal oj the general

population of various countries in the period 1955-67

Cminn\

Year

No.
Samples

DDT

(PPM)

DDE
AS DDT

(PPM)

Total
AS DDT

(PPM)

DDE
AS DDT

(%)

Reference

USA

1955

r

49

7.4

12.5

19.9

62.8

(11)

USA

1954-56

61

4.9

6.8

11.7

58.1

(12)

USA

l%l-62

130

4.0

8.6

12.6

68.3

(21)

USA

1963

28

2.4

4.2

6.7

62.0

(1)

USA

1963

282

2.9

8.2

11.1

73.9

(15, 16)

USA"

1955-56

16

2.3

3.6

5.9

61.0

(12)

USA'

1960

20

0.8

2.2

3.0

73.3

(5)

USA

1964

25

2.4

7.9

10.3

77

(14)

USA"

6

3.9

8.3

Fiserova-Bergerova et al

(1967)

USA«

10

7.2

12.7

Fiserova-Bergerova el al

(1967)

Canada

1959-60

62

1.6

3.3

4.9

67.0

(22)

Germany

1958-59

60

1.0

1.3

2.3

56.5

(18)

France

1961

10

1.7

3.5

5.2

67.3

(13)

Hungary

1960

50

5.7

6.7

12.4

51.3

(3)

England

1961-62

131

2.2

(17)

England

1964

100

4.0

(23)

India. Group I

1964

67

16.9

lO.I

26.0

39.0

(2)

India. Group 11

1964

19

20.3

10.7

31.0

34.0

(2)

India, Group III

1964

16

8.1

4.7

12.8

37.0

(2)

USA

1964-65

13

3,1-8.6

(24)

Czechoslovakia

1963-64

229

5.5

3.7

9.2

40.2

(9)

Israel

1963-64

254

8.5

10.7

19.2

55.6

(26)

Israel, Group I "

1965-66

71

4.6

5.6

10.2

52.1

This paper

Israel, Group II "

1965-66

133

8.2

9.9

18.1

53.9

This paper

' Persons eaiing no meal.

' Eskimos (Alaska).

• Age group 31-83, M.

* Farm workers, age group 18-57, M.

* Age group 0-9 years.

• Age group 10-89 years.

The further observations in this study support the con-
cept of placental transmission of DDT, which can be
considered a characteristic feature of pregnancy in our
epoch.

Neonates continue to receive DDT through their dietary
intake and probably in other ways too, since the storage
level of this insecticide does not diminish but, on the
contrary, increases with age.

Storage of high amounts of DDT-derived material can
occur in very early childhood. Thus, in this study, the
following amounts of total DDT have been found:
39.1 ppm in a female stillborn; 32.8 ppm in a day-old
neonate; 42.0 ppm in a male infant of 7 months and
60.0 ppm in a male infant of 1 I months. Our epidemi-

ological investigations carried out in order to clarify
the occurrence of these relatively high storage levels,
did not provide any meaningful explanation. In the
fat of two Indian children Dale et al. (2) found 291
ppm in a 3-year-old hoy and 180 ppm in a 7-year-old
girl. These authors attribute the high storage level of
these two children to the possibility of unusual exposure.

The population sampled for this study is exposed to
presumably similar environmental and living conditions,
but analysis of the storage of DDT-derived material re-
veals age and sex differences.

If one considers the results of the whole group 10-89
years, higher amounts of DDT, DDE. and DDT -|-
DDK are stored in comparison with the 0-9 year age
group (and particularly 0-2 years).

18

Pesticides Monitoring Journal

The results of this study also reveal a significant differ-
ence between the sexes in the storage of DDT-derived
material in the 10-89 year group, the men storing
more DDT and more DDE than the females. These
results accord with those of Hunter et al.. (17) who in a
sample of 1 3 1 necropsy fats in England found that the
total DDT concentration in specimens from males was
significantly higher than that in specimens from females.
In a further study on a sample of 100 (50 biopsy and
50 necropsy) specimens, Robinson et al. (23) found that
the mean DDT-derived material was 4.9 ppm in males
and 3.4 ppm in females. The mean concentrations of
p,p-DDE in the male samples, both biopsy and necropsy,
were significantly greater than those in the female
samples.

In this study sex differences are observed in all age
groups and even in the aged people in whom it is
thought that hormonal differences tend to disappear.
Higher storage of DDT-derived material in males could
be attributed to the fact that males generally eat larger
amounts of food than females.

In rats submitted to substantial dosage levels, Ortega
et al. (19) and Durham et al. (4) have found that the
female rat stores more DDT and much more DDE than
the male rat. Other species have shown small or no dif-
ferences between sexes in the storage of DDT: Woodard
et al. (28) in dogs, Harris et al. (10) in hogs, and Dur-
ham et al. (6) in monkeys.

Regarding the intensity of the population exposure to
DDT, the results of this study do not differ significantly
from those of our previous study (1963-64). As can
be seen from Table 3, the storage level of DDT-derived
material in the fat of people in Israel is persistently
high.

Summary and Conclusions

A total of 204 samples of human body fat were col-
lected in 1965-1966 from Israelis with no known occu-
pational exposure. The samples were analyzed by the
Schechter-Haller spectrophotometric method. Since the
method fails to distinguish between DDE and DDD, the
values given for DDE must be considered to represent
the sum of the two compounds. The mean total DDT-
derived material in 133 samples from persons aged
10-89 years is 18.1 ± 12.6 ppm: the mean for DDT
is 8.2 ± 6.1 ppm; and that for DDE is 9.1 ± 7.1 ppm.
DDE averages 53.9 ± 6.8% of the total DDT-derived
material. No significant difference was found between
the results obtained in this study and those of the
previous study (1963-64) on Israelis. There is no sig-
nificant difference for the DDT-derived material stored
in different age groups within the group aged 10-89
years.

In this group (10-89 years) males store higher amounts
of derived material than females, namely, DDT 9.3

Vol. 1, No. 2, September 1967

ppm versus 6.7 ppm; DDE 1 1.2 ppm versus 8.2 ppm;
DDT -|- DDE 20.5 ppm versus 14.8 ppm.

In the age group 0-9 years, the mean total DDT is
10.2 ± 9.2 ppm; DDT averages 4.6 ± 4.2 ppm; and
DDE 5.6 ± 5.9 ppm. DDE constitutes 53.3 ± 7.9%
of the total DDT-derived material. In this group of 71
cases, there were 12 stillborns, 15 neonates aged 0-7
days, 9 neonates aged 7 - 30 days, 33 infants aged 30
days - 2 years, and 2 older children. No significant dif-
ference was found in the storage of DDT-derived ma-
terial among stillborns, neonates, and infants.

There is a significant difference (p<0.001) in the
storage process of DDT-derived material between the
0-9 year group and that aged 10-89 years, as far as
the concentrations of DDT, DDE, and DDT + DDE
are concerned.

In the light of the data published during the past 10
years on the storage of DDT-derived material in the body
fat of people from various countries, this study supports
the -contention that DDT represents a current constitu-
ent of human body fat in the general population, and
that it is transmitted through the placenta to the fetus.
Its storage may present variations according to sex.
Further research is needed to clarify the mechanism of
this last feature.

This research study was supported by the U. S. Department
of Health. Education, and Welfare, Public Health Service,
National Communicable Disease Center, Atlanta, Georgia,
research grant No. BSS-CDC-IS-9.

LITERATURE CITED

(1) Dale, W. E. and G. E. Qumby. 1963. Chlorinated
insecticides in the body fat of people in the United
States. Science 142(3592): 593-5.

(2) Dale, W. £., M. F. Copclaud. and W. J. Hayes, Jr.
1965. Chlorinated insecticides in the body fat of people
in India. Bull. World Health Organ. 33:471-7.

(3i Denes. A. 1962. Problems of food chemistry concern-
ing residues of chlorinated hydrocarbons. Die Nahr.
6:48-56.

(4) Durham, W. F., C. Ciielo, Jr., and W. J. Hayes, Jr.
1956. Hormonal influences on DDT metabolism in the
white rat. Amer. J. Physiol. 187(2):373-7.

(5) Durham, W. F., F. F. Armstrong, W. M. Upholt, and
C. Heller. 1961. Insecticide content of diet and body
fat of Alaskan natives. Science 134(3493): 1880-1.

(6) Durham, W. F.. P. Ortega, and W. J. Hayes. Jr. 1963.
The effect of various dietary levels of DDT on liver
function, cell morphology, and DDT storage in the
rhesus monkey. Arch. Intern. Pharmacodyn 141(1-2):
111-29.

(7) Finnegan, J. K., H. B. Haag, and P. S. Larson. 1949.
Tissue distribution and elimination of DDD and DDT
following oral administration to dogs and rats. Proc.
Soc. Exp. Biol. Med., 72:357-60.

(8) Fiserova-Bergerova, V.. J. L. Radomski, J. E. Davics,
and J. H. Davis. 1967. Levels of chlorinated hydro-

19

1

carbon pesticides in human tissues. Ind. Med. Surg.
36(1); 65-70.
(9) llalacka. H.. J. Hakl. and F. Vy metal. 1965. Effect of
massive doses of DDT on human adipose tissue. Cesk.
Hyg. 10(3-4): 188-92.
110) Harris. L. E.. J. R. Harris. F. L. Mangelson. D. A.
Greenwood. C. Biddiilph. W. Binns. and M. L. Minncr.
1953. Effect of feeding DDT-treated alfalfa hay to
swine and of feeding the swine tissues to rats. J. Nutr.
51(4):491-505.

(11) Hayes. H'. J.. Jr.. IV. F. Durham, and C. Ciieto, Jr.
1956. The effect of known repeated oral doses of
chlorophenothane (DDTO in man. J. Amer. Med. Ass.
16:(9):890-7.

(12) Hayes. IV. J.. Jr.. C. E. Quinhy. K. C. Walker, J. W.
Ellioll. and W. M. Upholt. 1958. Storage of DDT
in people with different degrees of exposure to DDT.
Amer. Med. Ass. Arch. fnd. Health 18(5): 398-406.

(13) Hayes. W. J., Jr.. W. E. Dale, and R. LeBreton. 1963.
Storage of insecticides in French people. Nature
199(4899): 1189-91.

(14) Hayes. W. J., Jr.. W. E. Dale, and V. W. Burse. 1965.
Chlorinated hydrocarbon pesticides in the fat of peo-
ple in New Orleans. Life Sci. 4( 16): 161 1-15.

(15) Hoffman. W. S., W. I. Fishhein. and M. B. Andelman.
1964. The pesticide content of human fat tissue. Arch.
Environ. Health 9:387-94.

(16) Hoffman. W. S.. W. 1. Fishbein. and M. B. Andelman.
1964. Pesticide storage in human fat tissue. J. Amer.
Med. Ass. 188(9):819.

(17) Hunter, C. C. J. Robin.son. and A. Richardson. 1963.
Chlorinated insecticide content of human body fat in
Southern England. Brit. Med. J. 1:221-4. •

(18) Maicr-Bode. H. 1960. DDT in human body fat. Med.
Exp. 1:146-52.

(19) Onega. P.. W. J. Hayes, Jr.. W. F. Durham, and A.
Maitson. 1956. DDT in the diet of the rat. Public
Health Monograph No. 43, PHS, Publ. No. 484,1-27.

(20) Fillmore. R. £., /. O. Keith, L. C. McEwen, M. H.
Mohn. R. A. Wilson, and G. H. Ise. 1963. Cottontail
rabbit: feeding test. US Fish Wildlife Serv. Circ. No.
167.47-50.

(21) Quinby, G. E.. W. J. Hayes. Jr., J. F. Armstrong, and
W. F. Durham. 1965. DDT storage in the United
States population. J. Amer. Med. Ass. 191(3): 175-9.

(22) Read, S. T. and W. P. McKinley. 1961. DDT and DDE
content of human fat. Arch. Environ. Health 3:209-11.

(23) Robinson. J.. A. Richardson. C. G. Hunter, A. N.
Crabtree, and H. J. Rees. 1965. Organo-chlorine in-
secticide content of human adipose tissue. Southeast
Engl. Brit. J. Ind. Med. 22:220-9.

(24) Schafer, M. L. and J. E. Campbell. Distribution of
residues in human body tissues from Montgomery
County, Ohio. In: Organic Pesticides in the Environ-
ment. Edited by R. F. Gould, 1966, Advances in
Chem. Ser. No. 60, Amer. Chem. Soc, Wash., D. C,
p. 89-98.

(25) Technical Development Laboratories. 1953. Method
for the chemical analysis of DDT and DDE in samples
of fat. Chem. Mem. No. 1, Commun. Dis. Center,
USPHS, Savannah. Ga. (mimeogr.).

(26) Wasscrman. A/., M. Gon, D. Wa.ssermann, and L.
Zellermayer. 1965. DDT and DDE in the body fat of
the people in Israel. Arch Environ. Health 11(3): 375-9.

(27) Wassermann. M. and A. Hale. 1965. The role of air
pollution by insecticides as a body burden for the
general population. Proc. First World Congr. on Air
Pollution, Buenos Aires. (In press).

(28) Woodard. G.. R. R. Ofner. and C. M. Montgomery.
1945. Accumulation of DDT in the body fat and its
appearance in the milk of dogs. Science, 102:177-8.

(29) Zavon. M. R.. C. H. Mine, and D. K. Parker. 1965.
Chlorinated hydrocarbon insecticides in human body
fat in the United States. J. Amer. Med. Ass. 193(10):
837-9.

(30) Zavon. M. R.. R. Tyre, and L. Latorre. 1967. Chlor-
inated hydrocarbon pesticide levels in the newborn.
Proc. Con. Biol. Effects Pesticides Mammalian Syst.,
N. Y. Acad. Sci. (To be published).

20

Pesticides Monitoring Journal

RESIDUES IN FISH, WILDLIFE, AND

ESTUARIES

Insecticide Concentrations In Wildlife At Presidio, Texas

Dudley D. CuUey' and Howard G. Applegate=

ABSTRACT

Data are given on insecticide concentrations in representative
species of reptiles, birds, and mammals from Presidio, Texas.
Various tissues and organs were examined by the means of
gas chromatography. As specimens were taken farther from
the cultivated area, the insecticide concentrations decreased.
Within and adjacent to the cultivated area, insecticide con-
centrations showed a complex pattern. Possible reasons for
the complexity are discussed.

Introduction
PRELIMINARY studies on insecticides in an eco-
system at Presidio, Texas during June, July, and August
1965, have been reported in a series of papers (1-3,5,8).
This paper is an expansion of the paper by Culley and
Applegate (5) giving more data and reporting on an ex-
tended period of study.

The study was undertaken to provide training to students
in Departments of Wildlife, Plant Sciences, and Meteor-
ology in the detection of insecticides and to assess the
value of the Presidio Valley as an area in which to con-
duct long-term studies on the movement of insecticides
through an ecosystem and the effects and fate of insecti-
cides within the ecosystem. The data gathered during
the year must be regarded as preliminary and, perhaps,
indicative. They cannot be considered as conclusive.

Methods and Materials
The Presidio Valley has approximately 384.000 acres,
of which 2,900 acres are in cultivation. The area is geo-
graphically part of the Chihuahuaian Desert. The valley
supports excellent populations of a variety of reptiles,
birds, and mammals. The valley itself is at an elevation
of 2,600 feet, while the surrounding mountains in the
United States are above 7,000 feet, and in Mexico above
8,000 feet. Thus, we have essentially a point source of
insecticide application within a large enclosed area.
A map of the area is presented in Fig. 1 . Sites 1 and
4 were the westernmost and easternmost farms, respec-

tively, in the Presidio growing region. Site 2, located
between sites 1 and 4, was near the town. Each of these
sites (1,2, and 4) had two sampling stations: one in the
cotton fields and one in the desert peripheral to the
fields. Sites 7 through 9 were in the desert and located

UNITED STATES

3 3 MILES

1 Mississippi State University. State College, Mississippi.

2 Texas A & M University, College Station, Texas.

Vol. 1, No. 2, September 1967

FIGURE 1. Diagram of sampling area at Presidio, Texas.
Sites I 2 3 and 4 are in cotton fields; sites 5 and 6 are
water sampling areas; sites 7, 8, 9, and 10 are in the desert
and aerodynamicaily related to the Presidio region; site 11
is in the Chinati Mountains and site 12 north of the Chinati
Mountains.

21

3, 6, and 9 miles from the center of the growing region.
Site 12 (30 miles from the growing area) was located on
the north site of the Chinati Mountains and not normalh
related aerodynamically or hydrographicaily to the
Presidio Valley; this was used as the control area.

Representative specimens of reptiles, birds, and mam-
mals were obtained by shooting or trapping. Birds and
lizards were shot and placed on ice within an hour of
death. Mammals were trapped and killed by freezing.
All specimens were kept frozen until tissues were taken
for analysis. A minimum of four specimens per site of
each kind were collected on each date. Most of the
values presented represent mean insecticide concentra-
tions of five to six specimens.

Samples were analyzed by the method of Langlois,
Stemp. and Liska (9). Briefly, 1 g of frozen tissue was
ground with 20 g of Florisil. The sample-Florisil mix-
ture was placed on top of 25 g of partially deactivated
Florisil in a chromatographic column. Partial deacti-
vation of the Florisil was done by heating it to 90 C for
48 hours, adding 5'~r water (v/w) and then stoppering
tightly for 4<S hours. Insecticides were eluted from the
column with 1 liter of a mixture of 20% methylene
chloride in petroleum ether (v/v). The eluate was
evaporated to dryness, the residues taken up in 5 ml of
hexane and 5 ^1 injected into a gas chromatograph. The
gas chromatograph was an Aerograph 680 with an
electron capture detector. The column was 5 ft. x Vs in.
Pyrex and packed with 57c Dow-1 I on Chromasorb
W. The column temperature was 180 C, nitrogen pres-
sure was 20 psi, and the gas flow rate was 50 ml/min.
Thin layer chromatography was used to confirm the
identification of the insecticides.

All solvents used for extraction and chromatography
were purified by the methods of Burke and GiufTrida
(4).

Periodically, samples were spiked with all the insecti-
cides and their breakdown products for which data are
reported in this paper. The spiked samples were then
carried through the proper analytical procedure and
percent recovery of the compounds calculated. Re-
coveries ran from 83% to 96%.

In 1965, the first application of insecticides in the
Presidio Valley was to onions in March. This applica-
tion consisted of a fungicide (usually sulfur) and DDT.
Cantaloupes were sprayed with a fungicide and DDT
starting in May. Only small acreages were devoted to
these crops. The vast bulk of insecticides was applied
on cotton from late June to the middle of September.
During 1965, the followini; amoimts of insecticides (cal-
culated as pounds of the pure chemical) were applied
by the commercial growers: DDT - 20.750 lb,; methyl
parathion- 15.900 Ih.; Sevinf^ - 2,600 lb.; BHC - 2,585
lb.; ethyl parathion - 2,000 lb.; endrin - 200 lb. In

addition, seven sprays (three in late September and four
in October) of low volume, high concentration mala-
thion were applied under a Federal program. A total of
17,640 lb. of malathion was applied in the seven sprays
to the Presidio Valley.

Results

No differences could be detected in insecticide concen-
trations among three species of lizards — Cnemido-
phorus Tessellatus Say, C. r/VrA- B-ird and Girard, and
C. inornatus Baird. The data presented here are a com-
posite for all species. The data on insecticide concen-
trations in lizard tail muscle (Table 1), brain tissue
(Table 2), and liver tissue (Table 3), in general, show
that an increase in insecticide concentrations occurred /
during June, July, and August. There appeared to be
little difference in concentrations between samples from
the various sites in the cotton fields or from the desert
peripheral to the cotton fields. Concentrations decreased
up to 9 miles from the cotton fields. Thereafter they re-
mained essentially static.

Insecticide concentrations in post-coelomic fat bodies
of lizards are given in Table 4. Since many of the
samples contained no fat bodies, caution must be used
in drawing conclusions. There appeared to be a sharp
rise in concentrations from June through July followed
by a drop during August. In general, the concentrations
decreased up to 9 miles from the cotton fields. There-
after they remained essentially static.

The analysis of the stomach contents of the lizards
(Table 5) also showed that as specimens were gathered
at greater distances from the cotton fields, the insecticide
concentrations decreased. With the exception of DDE,
there were only slight increases in concentrations from
June through August. DDE showed pronounced in-
creases at all sites from July to August.

In the latter part of July many of the female lizards
that were collected contained eggs. Separate determina-
tions were made of the gravid female muscle tissue and
of the egg. In every case, the egg contained higher in-
secticide concentrations than did the gravid female
muscle tissue (Table 6). Then, concentrations in the
muscles of gravid females were compared with concen-
trations in the muscles of non-gravid females collected
on the same dates and at the same sites. No significant
differences could be detected.

The chemical names of compounds menlioned in this paper are:
BHC 1.2..^,4,5,6-hexachIorocyclohexane, mixed isomers

DDT I.I.I -irichloro-2.2-bis( p-chlorophcnyl ) ethane

TOE 1. 1 -difhloro-2.2-bis(p-chlorcphenyl) ethane

DDE I . I -dichloro-2.2-bis( p-chlorophenyl ) ethylene

Endrin 1 .2.:l.4.lll.l()-hcxachlo^o-6.7-epoxy-l.4.4a,.s.6.7.«,l^a-

oct,^hydro-l.4-<*f»/^»-(•/»/(;-5.S-dimell^a^onaphthalcne
Methyl Parathion o.rt-dimcthyl 0-;j-nitrophenyl phosphorothioate
Parathion ".^-diethyl ^'-;>-nitrophenyl phosphorothioate

Malathion dicth\l mcrcaplosiiccinate. .V-cstcr with O.rt-dinicthyl

phosphorodithioatc
Scvin'"' l-naphthyl mclhylcarbamatc

22

Pesticide.s Monitoring Johrnai

TABLE 1. — Mean insecticide concenirations (ppm) in lizard tail muscle; each sample is five to six tails

BHC

W
MP

Site 1
EST Cotton Field
P DDE TDE

DDT

BHC

Site 2

Center Cotton Field

MP P DDE TDE

DDT

BHC

Site 4

East Cotton Field

MP P DDE TDE

DDT

June

2.0

0.5

0.8 2.9

0.3

0.6

0.9

1.2

1.8 3.9

0.9

0.8

0.9

0.3

0.8 3.1

0.0

0,4

July

0.1

0.6

1.0 1.5

1.7

0.5

0.0

1.1

0.7 2.0

1.9

2.0

0.0

0.7

1.8 1,9

1.9

2,2

August

0.0

I.l

1.7 2.5

2.5

0.5

0.0

1.5

2.8 2.8

4.7

0.0

0.0

1.1

1.5 2.7

1.2

0.8

BHC

0.4

Wes
MP

n.9

Site 1
T Desert Periphery
P DDE TDE

DDT

BHC

Site 2
Center Desert Periphery
MP P DDE TDE

DDT

BHC

Site 4

East Desert Periphery

MP P DDE TDE

DDT

June

0.5 4.1

1.8

0.4

0.6

1.7

0.2 3.7

1.3

0.7

0.7

1.7

1.2 2.2

0.9

0,1

July

1.2

3.7

2.9 2.6

1.5

2.1

1.6

3.7

4.1 3.4

2.2

2.5

1.2

1.8

2.5 3.0

1.4

1.4

August

0.6

4.9

4.6 7.2

6.0

3.4

1.3

1.6

3.8 3.0

3.1

2.5

0.2

4.4

4.4 6.9

5.6

4.7

BHC

MP

Site 7
3 Mile Desert
P DDE

TDE

DDT

BHC

MP

Site 9

9 Mile Desert

P DDE

TDE

DDT

BHC

MP

Site 12
30 Mile Prairie
P DDE

TDE

DDT

June

0.2

0.6

0.3 0.9

0.4

0.6

0.3

0.1

0.0 0.1

0.1

O.I

0.1

0.1

0.0 0.1

0.2

0.2

July

0.6

1.2

1.0 1.9

1.1

1.4

0.2

0.6

0.4 0.3

0.2

0.4

0.0

0.1

0.1 0.3

0.1

0.1

August

0.5

1.5

2.4 2.5

1.4

1.7

0.4

0.6

1.1 1.1

1.0

0.4

0.2

0.7

0.8 0.6

0.4

0.3

TABLE 2. — Mean insecticide concentrations (ppm) in lizard brain tissi4e; each sample is five to si.\ brains

BHC

Site 1

West Cotton Field

MP P DDE TDE

DDT

BHC

Site 2

Center Cotton Field

MP P DDE TDE

DDT

BHC

E
MP

Site 4
AST Cotton Field
P DDE TDE

DDT

June

0.1

0.7

1.4 1.9

0.6

0.4

0.0

0.4

0.4 1.1

0.9

0,0

0.3

0.6

0.0 1.0

0.0

1.6

July

0.1

0.6

1.0 1.5

1.7

0.5

0.0

1.1

0.7 2.0

1.9

2.0

0.0

0.7

1.8 1.9

1.9

2.2

August

0.0

1.1

1.7 2.5

2.5

0.5

0.0

1.5

2.8 2.8

4.7

0.0

0.0

1.1

1.5 2.7

1.2

0.8

BHC

Site 1
West Desert Periphery
MP P DDE TDE

DDT

BHC

Site 2
Center Desert Perip
MP P DDE

hery
TDE

DDT

BHC

Site 4

East Desert Periphery

MP P DDE TDE

DDT

June

0.0

0.5

0.3 1.6

0.4

0,2

0.4

0.2

0.5 0.7

0.6

0.1

0.3

0.7

0.0 1.8

0.4

0.2

July

0.0

0.6

1.3 1.8

0.7

0.9

0.0

0.8

1.8 1.5

1.4

0.9

0.2

0.6

0.3 3.1

1.2

1.3

August

0.0

1.5

1.9 2.4

0.0

2.7

0.0

1.1

1.8 2.5

1.0

1.1

0.0

1.1

1.5 3.8

1.4

0,0

BHC

MP

Site 7
3 Mile Desert
P DDE

TDE

DDT

BHC

MP

Site 9

9 Mile Desert

P DDE

TDE

DDT

BHC

MP

Site 12
30 Mile Prairie
P DDE

TDE

DDT

June

0.0

0.2

0.4 0.3

0.6

0.1

0.0

0.0

0.0 1.5

0.5

1.1

0.0

0.1

0.3 0.3

O.I

0,2

July

0.0

0.7

0.6 0.8

1.2

1.2

0.0

1.2

1.1 0.6

1.2

1.6

0.0

0.1

0.9 0.6

0.4

0,1

August

0.0

1.0

0.7 1.6

1.4

0.8

0.0

0.4

1.8 0.7

1.5

0.6

0.0

0.1

0.9 0.8

0.0

0,0

TABLE 3, — Mean insecticide concentrations (ppm) in lizard liver; each sample is five to si.x livers

BHC

Site 1

West Cotton Field

MP P DDE TDE

DDT

BHC

Site 2
Center Cotton Fi
MP P DDE

eld
TDE

DDT

BHC

Site 4

East Cotton Field

MP P DDE TDE

DDT

June
July
August

0,0
1,0
0,0

1,1 0,6 1,0 1,2
1,0 0,8 2.1 1.6
1.5 1.2 3,7 2,0

0.3
0.6
1.3

0.0

I.l

0.1

1.0 1.3 2.1
0.4 0.8 3.3
1.0 1.1 3.4

1.3
1.6
0.9

1.0
0.5
0.6

0.0
0.3
0.0

0.2 0.1 2.5 0.1
0.4 0.4 1.5 0.6
0.9 0.6 3.8 2.1

0.2
0.6
0.7

Vol. 1, No. 2, September 1967

23

TABLE 3. — Mean insecriciile concentrations (ppm) in lizard li.er; each sample is five to six livers — Continued

BHC

Site 1
West Desirt Pehiphery
MP P nnF TDE

DDT

BHC

Site 2
Center Deseri Periphery
MP P DDE TDE

DDT

BHC

Site 4

East Desert Periphery

MP P DDE TDE

DDT

June

1.1

1 <

11 h .11

0.4

0.0

0.1

0.5

0.2 0.5

0.2

0.0

0.1

0.4

0.7 1.8

0.4

0.3

July

0.3

0,9

0.9 2.4

0.9

0.8

0,1

0.1

0,3 1,7

0.3

0.2

0.0

0.1

0.2 0.5

0.1

0.1

August

0.0

0.8

1.7 2.7

0.7

1.1

0.0

0.5

0.3 1.6

1.0

0.0

0.0

1.5

0.6 1.7

0.0

1.0

BHC

MP

Site 7
3 Mile Desert
P DDE

TDE

DDT

BHC

MP

Site 9
9 Mile Deseri
P DDE

TDE

DDT

BHC

MP

Site 12
30 Mile Prairie
P DDE

TDE

DDT

June

0.0

0.4

0.3 0.1

0.4

0.0

0.3

0.3

0.1 0.4

0.0

0.0

0.1

0.1

0.1 0.2

0.6

0.5

July

0.5

0.4

0.5 1.0

0.6

0.8

II.O

0.4

0,6 0.2

0.4

0.6

0.3

0.6

0.5 0.3

04

0.7

August

0.0

0.8

0.7 1.4

0.2

0.5

0.8

0.6

1.5 1.3

1.3

0.0

O.ll

0.5

0.1 0.5

0.1

0.1

TABLE 4. — Mean insecticide concentrations (ppm) in coelom fat of lizards: each sample is five to six cocloiii fat bodies

BHC

Sue 1

West Cotton Field

MP P DDE TDE

DDT

BHC

Site 2

Ci NTER Cotton Field

MP P DDE TDE

DDT

BHC

Site 4

East Cotton Field

MP P DDE TDE

DDT

'unc

No Data

1.5

0.0

0.3 31.5

19.2

14.5

No Data

July

11.5

4.2

2.8 17.4

34.3

43.0

8.9

0.0

1.4 45.9

32.0

44.3

No Data

August

8.5

2.1

1.0 21.2

20.8

25.1

1.2

0.0

0.0 18.8

7.6

7.6

0.4

0.3

0.6 5.8

3.4

4.0

BHC

Site 1
West Desert Periphery
MP P DDE TDE

DDT

BHC

Site 2
Center Desert Periphery
MP P DDE TDE

DDT

BHC

Site 4

East Desert Periphery

MP P DDE TDE

DDT

June

No Data

0.7

0.0

0.0 25.2

0.0

2.8

0.2

0.0

0.3 19.4

2.1

1.5

July

6.8

3.7

4.1 31.4

7.9

26.1

2.1

1.6

3.6 35.4

10.3

8.4

No Data

August

7.0

0.0

0.0 22.4

10.4

16.2

2.3

0.0

0.0 28.2

7.3

4.0

II. 1

0.1

0.1 17.7

0.7

1.9

BHC

MP

Site 7

3 Mile Desert

P DDE

TDE

DDT

BHC

MP

Site 9

9 Mile Desert

P DDE

TDE

DDT

BHC

MP

Site 12

30 Mile Prairie

P DDE

TDE

DDT

June

0.0

1.9

1.1 8.4

3.1

4.8

0.0

1.0

0.0 7.9

3.7

3.6

No Data

July

1.0

1.8

2.3 10.4

4.0

4.3

1.1

2.0

1.3 10.1

6.9

3.6

1.2

0.6

0.0 5.4

2.2

1.7

August

1.4

2.4

1.8 15.7

5.1

4.0

No Data

0.0

0.0

0.0 9.4

1.5

0.0

TABLE 5. — Mean insecticide concciUralions (ppm) of the stomach contents of lizards: each sample is five to six stomach

contents

BHC

Site 1

West Cotton Field

MP P DDE TDE

DDT

BHC

Site 2

Center Cotton Field

MP P DDE TDE

DDT

BHC

Site 4

East Cotton Field

MP P DDE TDE

DDT

June
July
August

0.0
0.6
1.5

1.1
0.8
1.4

0.7 0.9

1.5 2.0
1.3 7.7

0.6

2.7
1.4

0.0
4.2
1.3

0.1
0.6
1.1

0.9

0.6
1.7

0.8 1.2

1,1 2,7
1.3 4.7

0.7

1.8
1.5

0.2
1.4
1.6

0.1

0.0
0.2

0.4

0.5
1.9

0.2 1.3

0.9 1,7
1.7 3.4

0.2
0.6
0.9

0.2

0.3
0.7

BHC

Site I
West Desert Periphery
MP P DDE TDE

DDT

BHC

Site 2
Center Desert Periphery
MP P DDE TDE

DDT

BHC

SrrE4

East Desert Periphery

MP P DDE TDE

DDT

June
July
Audusl

0.0
0.0
0.0

0.6
0.3
0.8

0.6 0.4
0.4 0.2
1.3 4.5

0.4
0.4
0.2

0.3
0.4
0.0

0.0
0.0
0.3

0.6
0.2
1,0

0,4 0.6
0.1 0.3
0.3 3.2

0.4
0.3
0.2

0,0
0,3
0.5

0.1
0.0
0.3

0.9
0.3
1.7

0.2 0.4
0.4 0.5
0.5 1.9

0.1
0.3
0.8

0.1
0.2
0.4

BHC

MP

Site 7
3 Mile Desert
P DDE

TDE

DDT

BHC

MP

Site 9
9 Mile Desert
P DDE

TDE

DDT

BHC

MP

Site 12
30 Mile Prairie
P DDE

TDE

DDT

June
July
AuguM

0.0

0.0
0.1

0.4
0.2
0.6

0.2 0.4
0.3 0.9
0.4 1.5

0.4
0.3
0.3

0.2
0.3
0.4

0.1
0.0
0.0

0,1
0,5
0.5

0.1 0.2
0.1 0.4
0.4 1.0

0.2
0.3
0.2

0.1
0.2
0.2

0.1
0.0
0.0

0,1
0,0
0.1

0.1 0.1
0.2 0.2
0.2 0.5

0.0
0.1
0.1

0.0
0.0
0.1

24

Pesticides Monitoring Journal

TABLE 6. — Mean tnscclicide concentrations (ppm) in

muscle of four gravid and four non-gravid female lizard

species and eggs: all lizards were collected in tlie same field

BHC

MP

P

DDE

TDE

DDT

Females, gravid

1.1

2.5

1.4

3.4

2.8

2.1

Eggs

5.6

11.6

8.5

16.4

7.3

10.7

Females, non-gravid

0.8

2.3

2.1

3.4

2.8

2.0

Insecticide concentrations found in the breast muscle
(Table 7). brains (Table 8), livers (Table 9), and
gizzard contents (Table 10) of English sparrows
(Passer domesticiis L.) are given for specimens collected
in the cotton fields. Not enough birds were collected
from the desert to furnish reliable data. In general, in-
secticide concentrations increased from June through
August at all three sites. Birds collected from the west
end of the valley, however, had lower MP, P, DDT,
DDE, and TDE concentrations in their breast muscles
in August. Most samples contained less insecticides in
November than they did in August. There appeared to
be a slight rise in concentrations from November to
January for DDT, DDE, and TDE. Two species of
pocket mice {Perognathus penicillatus Woodhouse and
P. intermedins Merriam) and one species of kangaroo
rat (Dipodomys merriami Mearns) were used for
analysis. Approximately 2% of all mammals collected
were D. merriami. 8% were P. intermedins, and 90%
were P. penicillatus.

None of the above mammals could be found in the
cotton fields. They were collected in the desert less than
30 meters from the fields, however. In general, at all
sites on the periphery of the cotton fields insecticide
concentrations increased from June to July, remained
essentially static in August, and dropped in November.
In January an increase is apparent at all sites except
site 4 (Table 11) and the 9- and 30-mile stations
(Tables 1 1 and 12). Mammals collected from the desert
at the western periphery of the growing area had
greater insecticide concentrations than did those col-

lected from the desert peripheral to the center of the
growing area. Animals from the desert adjacent to the
easternmost cotton fields had the least concentrations of
insecticides. The farther the specimens were collected
from the cotton fields the less were the insecticide con-
centrations.

Discussion

No malathion was detected in any sample obtained
during the period of study reported in this paper. Since
some of the samples (those reported for November)
were obtained within 6 weeks after malathion applica-
tions, this indicates that the compound rapidly disap-
peared from the Presidio ecosystem.

Only a small amount of endrin was applied during the
1965 spraying season. It was detected in human urine
and cotton leaves, but not in leatherstem leaves, Rio
Grande water, or silt in the Rio Grande water (1,5,8).
Endrin was not detected in any of the reptiles, birds, or
mammals.

An attempt was made to obtain samples from each
class of vertebrates, except fish. Amphibians were not
found in sufficient numbers to sample during the
3-month period. From birds, the English sparrow was
selected due to both its numbers and year-round resi-
dence in the valley (13). The nearest areas in which
sparrows established residence were at Shafter and
Redford — 19 miles north and east of the valley, respec-
tively. It is unlikely there is interchange between these
three areas. P. penicillatus. P. intermedins, and D.
merriami were selected to represent the mammals due
to their small home range — 0.12 acres to 0.46 acres
(12.14). Reptiles were originally supposed to be repre-
sented by C. tigris. which has an estimated home range
of 0.26 acres (12). However, there were areas where this
species was not abundant. Therefore, C. tesselatus and
C. inornatns. which were abundant in such areas, were
also sampled. Milstead (//) has reported that all three
species have similar food habits and apparently occupy
similar ecological niches.

TABLE 7. — Mean insecticide concentrations (ppm) in breast iitiiscle of sparrows;
each sample is five to .■six breasts

BHC

Site 1

West Cotton Field

MP P DDE TDE

DDT

BHC

Site 2

Center Cotton Field

MP P DDE TDE

DDT

BHC

Site 4

East Cotton Field

MP P DDE TDE

DDT

June

0.0

2.0 1.7 5.2 4.1

3.4

0.0

3.2

2.0 4.4 2.7

1.9

0.3

2.7 3.9 1.6 2.7

3.1

July

1.7

4.4 6.5 7.0 8.1

5.0

0.0

3.5

2.7 8.0 6.0

5.0

0.6

3.6 3.2 4.5 2.6

4.6

August

1.9

2.1 1.5 6.8 4.6

3.2

1.9

4.9

5.3 16.2 10.2

7.1

0.9

5.5 3.4 11.8 7.8

6.6

Nov.

0.0

2.3 0.5 1.8 0.5

0.7

0.0

1.5

1.6 2.7 0.4

0.6

0.0

1.6 1.1 6.4 0.2

0.5

Jan.

0.5

4.3 2.5 3.9 1.5

0.9

0.3

2.9

2.0 5.7 0.4

1.2

0.0

1.9 1.0 7.2 1.2

1.0

Vol. 1, No. 2, September 1967

25

TABLE 8. — Mean iiiseclicitle concenlrations Ippml in hraiiis of sparrows: tiuli sample is five to six brains

BHC

Site I

West Cotton Field

MP P DDE TDE

DDT

BHC

Site 2

Center Cotton Field

MP P DDE TDE

DDT

BHC

Site 4

East Cotton Field

MP P DDE TDE

DDT

June

0.5

0.0 0.2 5.4 0.2

0.3

0.5

1,0 0 8 3.9 0.3

0.0

0.0

0.1 0.0 1.6 0.1

0.0

July

0.0

0.8 08 1.7 0.2

0.0

0.0

1.5 1.1 1.2 1.2

1.0

0.6

0.3 0.8 1.2 0.2

0.5

Ausust

2.0

0.9 0.7 6.3 0.0

0.5

1.7

1.3 0.9 5.0 0.5

0.5

0.2

0.7 0,9 1.8 0.8

0.5

Nov.

0.1

0.8 0.6 2.5 0.3

0.3

0.1

0.6 0.8 1.2 0.5

0.4

no

0.4 0.6 0.8 0.9

1.0

Jan.

It -

1 •; I.l 4.9 1.2

1.1

0.0

2.4 1.2 5.3 0.9

0.2

0.3

2.6 1.8 5.9 0.4

0.4

TABLE 9. — Mcim insecticide concenlialions (ppni) in liveis of sparrows: cacii sample is live to six livers

BHC

MP

Site 1

rsT CnrroN Field
P DDE TDE

DDT

BHC

Site 2

Center Cotton Field

MP P DDE TDE

DDT

BHC

Site 4

East Cotton Field

MP P DDE TDE

DDT

June

0.3

0.2

0.9 5.8 1.9

0.1

0.0

0.4 0.3 3.5 0.2

0.3

0.3

0.9 0.7 2.7

0.3

0.4

July

1.9

1.2

0,8 6.7 3.3

0.9

0.0

2.8 1.1 5,5 4,6

2.1

0.4

0.4 0.3 3.5

0.6

0.4

August

1,2

1.4

1.7 9.6 1.6

0.4

0.9

1.6 1.3 6.5 0,7

0.0

1)5

0.8 1,2 2.9

0.0

0.0

Nov.

0.0

1.0

0,5 4.6 1.1

0.8

0.4

1.0 1.7 3.5 0.9

0.4

0.0

0.5 0.3 1.4

0.3

0.3

Jan.

0.0

1.6

1.9 7.9 1.5

0.9

0,0

2.8 1.9 3.8 0.1

0.1

0.0

1.6 0.8 3.0

0.1

0.2

TABLE 10. — Mean insecticide concenlrations (ppm) in gizzards of sparrows: each sample is five to six gizzards

BHC

Site 1

West Cotton Field

MP P DDE TDE

DDT

BHC

Site 2

CtNTER Cotton Field

MP P DDE TDE

DDT

BHC

Site 4

East Cotton Field

MP P DDE TDE

DDT

June

0.5

0.9

0.6

2,1

0.0

0.4

no

0,3

0.3

2.3

0.6

0,5

0.0

0,4

0.2

2.6

0.2

0.2

July

0,1

1.2

1.1

3.2

0,0

0.3

0.2

3,2

0.8

5.4

1.5

0.8

n.l

0.7

1.3

3.6

0.3

0.3

AURUSt

11

1.7

1.8

8.6

1.0

0.0

1.9

0.8

2.5

6.5

0.0

0.3

n.o

1,6

1,3

4,1

0.0

0.3

Nov.

0,7

05

0.8

5,1

0,8

0.3

0.0

0.9

0.9

2,3

0.4

0.4

0.0

0,6

0.8

3.1

1.1

0,8

Jan.

0.4

0.7

1.2

5.8

0.5

0.9

0.0

0.9

0.5

2.7

0.3

0.3

0.2

0.9

0.2

1.3

0.5

0.8

TABLE 1 1. — Mean insecticide concenlrations (ppm) in leg muscles of pocket mice and
kangaroo rats: each sample is five to six leg muscles

BHC

SlTT 1
WpsT Desert Periphery
MP P DDE TDE

DDT

BHC

Site 2
Center Desert Periphery
MP P DDE TDE

DDT

BHC

Site 4

East Desert Periphery

MP P DDE TDE

DDT

June

0.3

:.<*

2,8 1,2

2.2

1,8

1.6

1,4

0.7 0.7

1.6

1.2

0.6

n,5

0.2 2.6

0.6

0.9

July

0.2

4.0

5,6 3.2

4,3

2.7

0.1

5,4

5.1 3.2

3.6

3.1

0.5

1.6

1,8 3.3

1.4

1.2

Aufiust

0 5

4.6

5.1 5.2

5.8

5.7

1.9

3.8

3.1 2.2

0.8

0.1

Lost

Nov.

0.0

0,7

0.9 0.8

1.1

1.2

0.1

1.3

0.9 2.6

0.3

0.2

0.4

1,1

1.2 3.3

0.7

0.7

Jan.

0.0

1.3

1.1 5.1

0,6

0.9

0.1

2.5

1.9 4.3

0.7

0.3

0.0

1.2

0.5 2.0

0.9

1.0

BHC

MP

Site?
3 Mile Desert
P DDE

TDE

DDT

BHC

MP

Site 9
9 Mm e Desert
P DDE

TDE

DDT

BHC

MP

Snx 12

30 Mile Prairie

P DDE

TDE

DDT

June

":

0.3

0.1 0.7

0.4

0.5

0.1

0.1

0,1 0.3

0.2

0.4

0.1

0.0

0.0 0.1

0.1

0.0

July

0.3

0.5

0.5 1.0

1.0

1.0

0.1

0.3

0.2 0.5

0.6

0.5

0.2

0.1

0.1 0.2

0.2

0.2

August

0.4

0.7

0.8 1.3

1.2

1.2

0.3

0.5

0.5 0.6

0,6

0.8

i),n

0.3

0.2 0.3

0.3

0.3

Nov.

0.1

0.5

0.7 0.3

0.2

0.3

0.2

0.6

0.8 0.4

0.1

0.5

on

0.9

0.4 0.2

0.3

0.4

Jan.

0.0

0.9

0.9 2.3

1.0

0.4

0.0

0.8

0.7 1.0

0.5

0.4

o,n

0.3

0.2 0.4

0.2

0.1

26

Pesticides Monitoring Journal

TABLE 12. — Mean inseclickle concenirations (ppmj in livers of pocket mice and kangaroo
rats: each sample is five lo six livers

BHC

SITC 1
West Desert Periphery
MP P DDE TDE

DDT

BHC

Site 2
Center Desert Periphery
MP P DDE TDE

DDT

BHC

Site 4

East Desert Periphery

MP P DDE TDE

DDT

June

0.1

0.9

0.8 2.6

0.9

1.7

0.2

0.6

0.7 2.1

0.7

0.7

0.0

0.4

0.6 1.9

0.4

0.8

July

0.1

1.4

1.2 0.8

1.0

0.8

0.1

0.7

0.5 2.6

0.4

0.8

0.0

1.1

r.2 2.9

0.8

0.7

August

03

1.2

1.5 4.2

0.3

0.9

0.0

1.1

1.2 3.7

0.6

0.9

Lost

Nov.

0.0

0.4

0.7 1.2

0.4

0.4

0.0

0.8

0.9 1.2

0.3

0.3

0.0

I.I

0.8 2.4

0.7

0.5

Jan.

0.0

1.1

0.6 2.9

0.2

0.9

0.3

1.7

1.9 2.8

2.0

1.2

0.8

1.9

1.8 2.9

3.4

1.5

BHC

MP

Site 7

3 Mile Desert

P DDE

TDE

DDT

BHC

MP

Site 9

9 Mile Desert

P DDE

TDE

DDT

BHC

MP

Site 12

30 Mile Prairie

P DDE

TDE

DDT

June

0.2

0.6

0.7 0.8

0,5

0.1

0.0

0.2

0.4 0.3

0.7

0.3

0.0

0.2

0.2 0.3

0.2

0.0

July

0.3

0.8

0.6 1.3

0.5

0.4

0.0

0.7

0.4 0.6

0.1

0.5

0.0

0.3

0.4 0.2

0.4

0.2

August

0.3

0.9

0.7 1.8

0.3

0.7

0.0

1.0

0.5 1.3

0.4

0.5

0.0

0.7

0.2 0.3

0.5

0.1

Nov.

O.I

0.7

0.4 0.9

0.5

0.3

0.5

0.9

0.4 0.8

0.5

0.4

0.0

(1.1

0.3 0.3

0.2

0.2

Jan.

0.0

0.8

0.6 1.3

0.6

0.5

0.1

0.1

0.4 0.6

0.9

0.0

0.0

0.1

0.3 0.6

0.4

0.2

It should be noted that I 5,900 lb. of methyl parathion
and 2,000 lb. of ethyl parathion were sprayed in the
Presidio Valley in 1965. This is an 8:1 ratio. However,
an inspection of our data shows that this ratio of methyl
parathion to ethyl parathion was not found in either the
biological material or in the soil and water. The reason
for the unbalance between the ratios applied and the
ratios detected is not apparent. In tail muscle, brain,
and liver tissues of the lizards, BHC concentrations de-
creased from June through August. During this same
period, MP, P, DDT, DDE, and TDE increased in con-
centration in these tissues. With the exception of DDE
(insecticide concentrations in the stomach contents in-
creased only slightly in the same period. DDE residues
in the stomach contents showed pronounced increases
during this period. The stomach contents of lizards from
the cotton fields consistently had higher insecticide con-
centrations than did the stomach contents of lizards
collected adjacent to the fields. An examination of the
stomach contents showed that lizards in the cotton
fields ate mainly grasshoppers, while those in the desert
ate mainly termites. The grasshoppers were more
directly exposed to insecticide applications than the
termites. In addition, the grasshoppers' diet of fresh
foliage as contrasted to the termites' diet of organic
debris could also account for the differences in stomach
concentrations between the two lizard populations.

Insecticides in the coelomic fat increased sharply from
June to about the first of August in lizards from the
cotton fields. Thereafter, they dropped greatly in con-
centration until, at the end of August, the concentra-
tions were, in many cases, less than at the end of June.

However, the coelomic fat even in August contained
greater concentrations of chlorinated insecticides than
did the muscle, liver, brain, and stomach contents. The
r.torage of chlorinated insecticides in the fat of mammals
has been well documented. It was not surprising to find
that reptiles also store these compounds in fat.

Whiptails in the Presidio area hibernate from Septem-
ber to May (Milstead. personal communication and our
observations). The adults of any given year were born
the previous year. Breeding appears to take place during
the entire summer (11). In contrast to Milstead, who
found mature eggs complete with shell in early June,
we did not find any eggs until the latter part of July.
These eggs were small and immature. Milstead (personal
communication) believes that a post-coelomic fat body
is used in egg development. Hahn and Tinkle (6) re-
ported that post-coelomic fat may serve a reproductive
function with Uta stanshuriano, a species living in a
habitat similar to C tesselatus and tigris. Hoddenbach
(7) confirmed this in female race runners (C. sexlineatus)
in western Texas. It appears likely that the post-coelomic
fat body in Cnemidophorus spp. has a similar reproduc-
tion function. This being the case, insecticide concen-
trations in coelomic fat would be transported directly to
the developing egg. The peak concentrations in the
coelomic fat occurred in late July — concomitant with
the appearance of eggs.

Tinkle (15) and Maslin (10) have reported on the preva-
lence of females in C. tesselatus. We found two imma-
ture males in 3 months' collecting in 1965 and one
mature male in 2 weeks' collecting in 1966. All three

Vol. 1, No. 2, September 1967

27

specimens were collected adjacent to the cotton fields.
Only one male, C. tesselalus. has heen reported previ-
ously (10).

There is a much closer correlation between insecticide
concentrations found in the food taken from the gizzard
of sparrows and their tissue concentrations than between
the insecticide concentrations in the stomach contents
of lizards and their tissue concentrations. As the spar-
row tissue varied in insecticide concentration from date
to date and from site to site, the gizzard content concen-
trations varied in a similar manner. The variations in
insecticide concentrations in all tissues and food from
site to site are difficult to assess due to lack of informa-
tion concerning movement, behavior, feeding habits,
and food quality of the sparrows and lizards.

Concentrations in samples fell in November (after all
spraying had stopped) as would be expected. The
slight rise noted in January may or may not be signif-
icant, but appeared in mammals as well as birds. The
use of fat reserves might cause such a rise. Further
work is needed to clarify this point.

Due to the use of bait, no concentrations in stomach
contents could be obtained for mammals. In general,
the variations for the mammals followed the variations
observed in the birds rather than those of lizards. Varia-
tions of insecticide concentrations from site to site were
more closely correlated with changes in soil and vegeta-
tions (2.8) than were changes in the birds and lizards.

There were cases where methyl and ethyl parathion
residues in June exceeded those in July in reptiles, birds,
and mammals. Applegate (2) reported that leaves of
leatherstem (a perennial) had higher methyl and ethyl
parathion concentrations in June than did leaves of cot-
ton (an annual). This was interpreted as an indication
that these insecticides could accumulate in leatherstem.
It would appear, in the Presidio area, that reptiles, birds,
and mammals can also accumulate methyl and ethyl
parathion from one spraying season to the next spraying
season.

It is apparent that very complex interactions exist in
the movement of insecticides through the air to soil,
plants, water, insects and, ultimately, into reptiles, birds,
and mammals. More information is needed about the
concentrations of insecticides present at various levels
in food chains. The entrance of insecticides into organ-
isms as mediated by the organisms' shelters, movements,
and behavior is not presently available.

Technical Article No. 5762, Texas Agriciihural E.xpeiimeiil
Sinlion. College Stalion. Texas. This research was supported
in pan by Grant AP 28-02 from the Division of Air Pollu-
tion, Bureau oj State Services, Public Health Service.

28

Acknowledgments

We wish to thank Dr. W. B. Davis, Department of Wild-
life Science, Texas A & M University, for aid in identifi-
cation of the mammals and Dr. T. Paul Maslin, Curator
of Herpetology, University of Colorado Museum,
Boulder, for aid in identification of the lizards. We
appreciate the critical reading of this paper by Dr.
Denzel E. Ferguson. Department of Zoology, Mississippi
State University and Dr. Jack Inglis, Department of
Wildlife Sciences, Texas A & M University.

LITERATURE CITED

It) Applegate. H. G. 1966. Pesticides at Presidio I. General
survey. Tex. J. Sci. 18:171-178.

(2) Applcs;alc, H. G. 1966. Pesticides at Presidio II. Vege-
tation. Tex. J. Sci. (Accepted for publication).

(3) Applegate, H. G. 1967. Pesticides at Presidio V. Sum-
mary. Tex. J. Sci. (Accepted for publication).

(4) Burke. J., anil L. Giufjriila. 1964. Investigations of
electron capture gas chromatography for the analysis
of multiple chli>rinated pesticide residues in vegetables.
J. Ass. Agr. Chem. 47:326-342.

(5) Cullcy. D. D.. and H. G. Applegate. 1967. Pesticides
at Presidio IV. Reptiles, birds, and mammals. Tex. J.
Sci. (Accepted for publication).

(6) Hahn, W. E., and D. W. Tinkle. 1965. Fat body cycl-
ing and experimental evidence for its adaptive signifi-
cance to ovarium follicle development in the lizard
Vta slansburiana. J. Exp. Zool. 158:79-86.

(7) Hoddenhach, G. A. 1966. Reproduction in Western
Texas Cnemidophorus sexlinealus (Sauria: Teiidae).
Copeia 1:110-113.

(8) Laliser, C, and H. G. Applegate. 1966. Pesticides at
Presidio III. Soil and water. Tex. J. Sci. (Accepted
for publication).

(9) Langlois, B. E.. A. R. Stemp. and B. J. Liska. 1964.
Analysis of animal food products for chlorinated in-
secticide residues I. Column clean-up of samples for
electron capture gas chromatographic analysis. J. Milk
and Food Tech. 27:202-204.

(10) Maslin, T. P. 1962. All-female species of the lizard
genus Cnemidophorus, Teiidae. Science 135:212-213.

(11) Milslead, W. W. 1957. Some aspects of composition
in natural populations of whiptail lizards (genus
Cnemidophorus). Tex. J. Sci. 9:410-447.

(12) Milstead, W. W. 1961. Observations of the activities
of small animals (Reptillia and Mammalia) on a
quadrat in southwest Texas. Amcr. Midland Natur.
65:127-138.

(13) Peterson, R. T. 1963. A field guide to the birds of
Texas. Houghton Mifflin Co. Boston. 304 p.

(14) Reynolds. H. G. 1960. Life history notes on Merriam's
Kangaroo rat in southern Arizona. J. Mammalogy.
41:48-58.

(15) Tinkle. D. W. 1959. Observations on the lizards
Cnemidophorus tigris, Cnemidophorus te.i:setalus and
Crotaphytus wistizeni. Southwestern Natur. 4:195-200.

Pesticides Monitoring Journal

An Evaluation of the Effects of the Aedes aegypti Eradication
Program on Wildlife in South Florida'

Philip N. Lehner, Thomas O. Bosweli", and Frank Copeland'

Introduction

THE objective of this study was to evaluate the effects
of the Aedes aegypti Eradication Program in South
Florida on Wildlife other than Aedes aegypti. the target
organism. The field investigations and collection of
specimens for this evaluation were carried out during
the period May 10 to August 27, 1965.

The authors were furnished a selected group of written
complaints from among those received by the Aedes
aegypti Eradication Program in Atlanta, Georgia. These
complaints were then categorized by time and type.
Newspaper and magazine articles were reviewed to de-
termine the extent and type of problems inciting public
antagonism. Naturalists, civic leaders, and personnel
conducting the Aedes aegypti eradication operations
were interviewed personally or by telephone.

Sick and dead birds of various species were collected
from the treated areas, and healthy house sparrows
and mockingbirds from both treated and untreated
zones. Bird eggs also were collected. Specimens were
shipped at intervals to the National Communicable

' From the U. S. Department of Health, Education, and Welfare,
Public Health Service, Bureau of Disease Prevention and Environ-
mental Control. National Communicable Disease Center, Aedes
aegypti Eradication Program, Atlanta, Georgia 30333.

- Presently graduate research assistant with the Utah Cooperative
Wildlife Research Unit, Utah State University, Logan, Utah 84321.

^ Presently graduate research assistant with the Department of Biol-
ogy, New Mexico State University, University Park, New Mexico
88070.

* Formerly Analytical Chemist, Toxicology Section, Technology
Branch, Communicable Disease Center, Atlanta, Georgia; present
address. NCDC Pesticide Research Laboratory. P. O. Box 490,
Perrine, Florida 33157.

Disease Center, Toxicology Laboratory, Atlanta,
Georgia, to be analyzed for chlorinated hydrocarbon
insecticides. Pathological analyses were conducted on
some specimens by the Animal Diseases Diagnostic
Laboratory, Miami Section, Florida Department of
Agriculture. In addition, bioassay tests were made by the
Diagnostic Laboratories Section, Division of Animal
Industry, Florida Department of Agriculture, Kissim-
mee, Florida.

Field Investigation

OBSERVATIONS OF SPRAY OPERATIONS

The Dade County spray operations of the Aedes aegypti
Eradication Program are conducted on a zone basis.
The zones generally are those delineated for city census
purposes. There is a great variance in the size of zones;
they usually contain 50 to 100 blocks and hundreds of
premises. The premises likewise vary widely in size,
ranging from individual home sites to large parcels of
vacant land, some as large as 20 acres.

Before spraying is begun, the zones are checked by in-
spectors to determine the number of premises in the
zone that contain Aedes aegypti larvae. From these data
are derived two indices: (1) block index — the percent
of positive blocks in the zone — and (2) premises index —
the percent of positive premises in the zone.

Using these indices, a decision is made as to how the
zone will be sprayed. Following are definitions of the
three degrees of application employed.

Vol. 1, No. 2, September 1967

29

1 . Comprehensive Spraying — The treatment of all
breeding and potential breeding containers and the
area immediately around these containers on all
premises of all blocks in a given area. In areas
where excessive vegetational growth precludes posi-
tive detection of hidden containers, spray applica-
tions will be applied.

2. Encompassmeni Sprayinfi — The treatment of all
breeding and potential breeding containers and the
area immediately around these containers on all
premises in infested blocks and in the blocks im-
mediately adjacent to infested blocks.

COLLECTION OF SPECIMENS

Seventy-four specimens — 55 birds and 19 bird eggs —
were collected by the authors in the Miami area. Of
the 55 bird specimens, 41 were taken in healthy con-
dition, 7 were found sick and later died, and 7 were
dead when found. Four additional bird specimens were
received from a member of the Florida Audubon Society
in West Palm Beach. Of these one was found dead
following spraying, and the other three were found
sick and ultimately died. Unfortunately, all of the eggs
collected were in such poor condition upon arrival at
the Atlanta laboratory that analysis for insecticide
content was not practical.

3. Spot Treatments — This type of treatment refers
to a special situation, for example, application of
insecticides to bromeliads or to areas around fish
ponds where routine spraying might be impractical.

Besides the various degrees of application employed,
there are three basic methods of application: spraying
by truck-mounted power sprayers, spraying by hand
compression sprayers, and dispensing of dust by hand
equipment. The decision as to method of application
in specific situations must often be made by spraymen,
using general guidelines provided by the area super-
visors.

The spray formula used during this study was a xylene-
water emulsion containing 1.25% DDT by weight.
Spray used during 1964 and early 1965 was 2.50%
DDT. The 1.25% spray contains approximately 0.1 lb.
of DDT/gallon of spray.

A total of 64 hours was spent observing spray opera-
tions. The following is a list of the spray applications
that were inconsistent with operational standards and
represent a hazard to wildlife:

No. of Observations Misuse of Spray

9 Spraying areas obviously clean of

containers
5 Blanket spraying

3 Spraying pond or waterway

3 Carelessness with equipment result-

ing in excess spray deposits

Apparently there was not only great inconsistency in
spray application between and within spray crews, but
also nearly complete lack of knowledge as to prescribed
spray procedures. This can be partly accounted for by
Ihc numerous individual circumstances that arise in the
field: however, there was also a great difference observed
in the treatment of similar objects, such as birdbaths.
animal dishes, shrubbery, and ornamental plants.

Laboratory A nalysis of Specimens

All of the bird specimens were analyzed for brain levels
of chlorinated hydrocarbons. Also, pathological analyses
were made on 11 of the 14 specimens found sick or
dead. Tables 1, 2, and 3 present data for the 59 speci-
mens analyzed. For the purposes of this study, only the
levels of DDT and its metabolites have been listed.
Previous work at the Patuxent Wildlife Research Center
suggests that the brain level of DDE is not a good indi-
cation of lethality (1).

In the laboratory, the bird brain samples were ground
with sodium sulfate and then extracted with 25 ml of
nano-grade «-hexane for 1 hour with the aid of a wrist
action shaking machine. After extraction, the samples
were filtered through a small plug of glass wool into
50-ml test tubes. The solvent was then evaporated down
to 4 ml in a 40 C water bath with the aid of a gentle
stream of clean dry air. As a clean-up procedure the
4-ml hexane extract was partitioned three times with 4
ml of nano-grade acetonitrile which had been equili-
brated with hexane (1:1). The acetonitrile phases were
then combined and evaporated down to 1 ml as de-
scribed above. Two ml of distilled water were added,
and the acetonitrile-water phase was extracted three
times with 2-ml portions of nano-grade hexane. The
hexane extracts were dried with sodium sulfate and
then combined in a 15-ml centrifuge and evaporated
down to 0.2 ml and appropriate aliquots subjected to
gas chromatography.

A Microtek 2503R gas chromatograph equipped with
a microcoulometric detector and a strip chart recorder
was used. In addition to the microcoulometric detector
which is specified for chlorine, two columns were also
used to confirm the identity of the materials in the
eftlucnt gas. Both columns consisted of an aluminum
tube 6 ft. X Va in. O.D. Column No. I was packed with
2.5% diethylene glycol succinate (D.E.G.S.) on 60/80
mesh, acid-washed chromosorb G. Column No. 2 was
packed with .3% QF-1 on 60/80 mesh acid-washed
chromosorb Ci. Both columns were operated under the

30

Pesticides Monitoring Journal

following conditions: inlet and outlet blocks 230 C;
column oven 170 C; transfer line 230 C; combustion
furnace 800 C; carrier gas N^ 60 cc/min; oxygen 100
cc/min; bias 250 mv. The retention times in minutes of
columns 1 and 2, respectively, were as follows: p.p'-
DDT, 24.0 and 33.8; o.p'-DDT, 12.8 and 22.4; p.p'-
DDE, 9.2 and 17.6; o,p'-DDE, 7.2 and 13.4; p./p'-DDD,
30.5 and 31.2; alpha-BHC, 3.8 and 5.6; beta-BHC,
18.9 and 8.3; gamma-BHC, 5.9 and 7.0; delta-BHC,
17.1 and 9.2; heptachlor epoxide, 8.0 and 16.3; dieldrin,
11.9 and 25.4.

Results and Discussion

Tables 1 through 3 report "less than" values in order
to give the reader an idea of the sensitivity of our
method. The "less than" values show variation because
the sample sizes and therefore the weight represented by
any given aliquot varied. The "less than" values give
more information to the reader than a simple "not de-
tectable" notation. It was also felt that to show a "zero"
would have been false. The authors believe that with
larger samples or with more sensitive detectors, the
number of positive readings could have been increased.

Since gas chromatography is a more sensitive technique
than paper, thin layer, or infrared, and since we were
not able to detect anything by gas chromatography in
many samples, pesticides would not have been detected
by these less sensitive methods. Therefore we resorted
to the use of two columns of different polarity and the
microcoulometric detector which is specific for halogens
as reasonable proof of identity of the compounds in the
effluent gas.

Tables 1 and 2 show the results of analyses of 33
nestling house sparrows, 1 8 from zones treated twice
with DDT and 1 5 from untreated zones. House sparrows
obtained from treated zones were collected at least a
block inside the periphery of the zone. It is immediately
apparent that there is no observable difference between
the insecticide levels in brain tissue found in these two
sets of samples. The insecticide levels for Specimen No.
29 were much higher than for the remainder of the
population from the treated area, indicating that al-
though this bird was probably not killed by DDT, it had
received amounts far above what would be expected in
that zone.

Treatment of Zone 9C for the fifth time was begun
just previous to termination of the study and did not
allow sufficient time for thorough investigation. Because
Zone 9C was receiving treatments greatly in excess of
other zones observed, five specimens were collected from
it (four alive and one dead).

Specimen No. 68 was a young domestic turkey allowed
to run loose in a yard in Zone 9C. It was taken inside the
house while the premises were sprayed in the morning,
but was released into the yard again that same afternoon

Vol. 1, No. 2, September 1967

and died by mid-afternoon. The owner's description of
the turkey's death suggested loss of motor control and
periodic muscle spasms indicative of neurotoxic poison-
ing. The brain level of DDT + DDD was 21.82 ppm.
Although a few birds have been known to die of DDT
poisoning with brain levels this low, the level does not
approach the tentatively accepted minimal lethal brain
level of 30 ppm (/). Circumstantial evidence indicates
DDT poisoning but is not fully supported by results of
brain analysis in light of the available knowledge today.

Specimen No. 75 was an adult loggerhead shrike col-
lected in Zone 9C to serve as an indicator of contam-
ination of the food chain, since shrikes are almost ex-
clusively carnivorous and insectivorous. The shrike gets
most of its food from one trophic level higher than
songbirds. The high level of DDE and very low level of
DDT + DDD found in this specimen suggest a long-
term buildup of DDE from the environment but little
recent exposure to DDT. Present available knowledge
does not permit an interpretation of the significance of
this level of DDE; however, this shrike was apparently
healthy when collected.

Three dead birds (a duck, a mockingbird, and a coot)
were received by the Diagnostic Laboratories Section,
Florida Department of Agriculture, from cities sprayed
by the Program. Results of bioassay tests made on these
birds were all negative. There was no evidence, however,
that these birds were from actual sprayed areas within
the cities.

Four specimens. Nos. 69-72, received from a resident
of West Palm Beach (Resident No. 1) on August 12,
1965, were alleged to have been killed by heavy spray
applications made by the Aedes aegypti Eradication
Program in late 1964 and early 1965. Specimen No.
69 (myrtle warbler) was found dead by this individual
at her residence 2 days after the surrounding premises
had been sprayed. Brain analysis showed that death
cannot be attributed to DDT poisoning.

Specimen Nos. 70-72 were collected by another resi-
dent of West Palm Beach (Resident No. 2), who froze
each of them after death and sent them with an accom-
panying letter describing the deaths to a third resident of
the city (Resident No. 3). The available information
indicates that Resident No. 3 then sent the specimens to
Resident No. 1, who kept them frozen until she turned
them over to the authors on August 12.
Brain analysis of specimen No. 70 (crow) showed a
sizeable quantity of DDE but only small amounts of
DDT and DDD. This indicated either a heavy exposure
to DDE through the food chain or a past heavy exposure
to DDT and/ or DDD and their metabolism to DDE and
storage in the bird.

The brain level of DDT + DDE in specimen No. 71
(cardinal) was 27.31 ppm. Thus, it is probable that the

31

TABLE 1. — Pesticide analyses of brain tissue (ppm) from nestling sparrow colh'clions from
zones treated twice in 1965 with 1.25% DDT

Condition

WHEN
COLLECTED

Date
collected

Date of

LAST

treatment 1

ACCREGATB

Ave. Gal./
Premises =

Pesticide levels in brain issue (ppm)

No.

DDT

DDD

p,p'-DDE

o,p'-DDE

2

Healthy

5/11

4/22

4.92

< .58

< .30

.40

< .30

6

Healthy

5/1 J

4/22

4.92

<1.16

< .58

< .58

< .58

7

Healthy

5 '14

4/22

4.92

.63

.25

1.07

< .24

8

Healthy

5/14

4/22

4.92

< .78

.39

.25

< .39

9«

Healthy

5/14

4/22

4.92

<1.08

< .54

1.56

< .54

10 ♦

Healthy

5/14

4/22

4.92

< .88

< .44

2.67

< .44

26

Healthy

6/24

6/18

11.78

< .44

< .22

.33

< .22

27

Healthy

6/24

6/18

11.78

< .50

< .25

.86

< .25

28

Dead

6/24

6/18

11.78

<1.36

< .68

1.50

< .68

29

Dead

6/24

6/18

11.78

2.81

< .54

5.68

< .54

33 •

Healthy

7/6

6/6

6.06

.55

< .86

1.19

< .86

37

Healthy

7 14

6/6

6.06

< .88

< .44

.56

< .44

38

Healthy

7/14

6/6

6.06

< .60

< .30

.12

< .30

39

Healthy

7/14

6/6

6.06

.31

< .53

.26

< .53

40

Healthy

7/14

7/28

1.46

< .53

< .26

.65

< .26

41

Healthy

7/14

7/28

1.46

.18

< .30

.57

< .30

46

Healthy

7/20

6/6

6.06

< .88

< .44

.21

< .44

47

Healthy

7/22

6/6

6.06

< .85

< .43

.66

.19

* Type of treatment: Comprehensive.

3 The sum of the two averages, one for each treatmenl.

* Sample contains three nestlings.
« Sample contains five nestlings.

^ Sample contains four nestlings.

TABLE 2. — Pesticide analyses of brain tissue (ppm) from nestling house sparrow collections
fron} untreated zones, 1965 {healthy}

Specimen

No.

Date
collected

Approx. distance

(miles) to nearest

treated zone

Pesticide levels in brain tissue (ppm)

DDT

DDD

p,p'-DDE

o,p'-DDE

19

6/15

13

.34

< .62

.39

< .62

20

6/15

13

< .82

< .41

.22

< .41

21

6/15

13

<1.07

< .53

.66

< .53

22

6/16

7

< .36

< .18

.25

.10

23

6/16

7

< .38

< .19

.28

< .19

24

6/16

7

< .37

< .19

.16

< .19

SO

7/22

4

< .50

< .25

.72

< .25

SI

7/22

4

< .70

< .35

.81

< .35

52

7/22

4

< .62

< .31

.43

< .31

53

7 '22

4

< .91

< .45

.70

< .45

55

7 23

10

<2.04

<1.02

.32

<1.02

56

7/23

10

< .74

< .37

.18

< .37

57

7/23

10

.80

< .47

.33

< .47

S8

7. '23

10

< .79

< .40

.25

< .40

59

7 23

10

< .57

< .28

.18

< .28

32

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Vol. 1, No. 2, September 1967

.= o —

33

bird died of DDT poisoning: if it did not. it certainly
was very close to reaching a lethal brain level. Cardinals
were most often mentioned in reports of past wildlife
damage. Their disappearance particularly was related to
spra\ing. although some people mentioned finding dead
cardinals. Specimen No. 71 was the only cardinal that
was saved during the period when wildlife damage was
supposed to be greatest. No cardinals were collected
during the present study period. A letter dated Decem-
ber 16, 1964, which accompanied the dead bird when
it was submitted as a specimen, stated that when the bird
was collected, it could not balance itself but repeatedly
fell on its back until its death. This description of the
bird's death would fit several types of poisoning wherein
the organism loses motor control. It does not include a
description of the tremors that accompany DDT poison-
ing and are usually obvious to the observer. This bird
was collected less than 13 days after the area was
.sprayed, which is a reasonable time lapse in which to
expect detrimental effects to wildlife to appear.

Brain analysis of the ground dove (specimen No. 72)
collected by the same resident showed that this bird was
carrying 52 ppm of DDT ^ DDD, an amount well in
excess of what is tentatively considered to be the lower
lethal level. This bird was collected 8 days after the
zone was sprayed. In her accompanying letter of De-
cember 21. 1964, the collector described the bird's
death as being accompanied by uncontrolled twitching
and convulsive movements of the feet. This description
is consistent with the symptoms of DDT poisoning,
which are similar, of course, in any neurotoxic poison-
ing. With the combination of high brain level of DDT -f
DDD and the 8-day time lapse after spraying, it can be
stated with some certainty that this bird died of DDT
poisoning — circumstantial evidence indicated that the
Aedes aegypii Eradication Program could have been
the source of the DDT.

Conclusions

No evidence was found of mass kills of vertebrates that
could be attributed to the Aedes aegypti Eradication
Program. Because of the operational methods employed
by the spray program during this investigation (with the
exception of Zone 9C), there was little reason to suspect
immediate and widespread damage to wildlife. By far
the greatest number of personal complaints was directed
at the use of DDT and not first-hand accounts of wild-
life damage.

Analyses of specimens from West Palm Beach tended
to support claims of some wildlife damage reported
from that area before this investigation. Brain analysis
of a ground dove indicated a high probability that this
bird was killed by DDT poisoning. In one cardinal, the
brain level of DDT + DDD was high enough to seri-
ously endanger the bird, if not to cause its death.

34

Circumstantial evidence in these cases indicated that
the Aedes aegypti Eradication Program could have been
the source of the DDT. One crow showed heavy ex-
posure to DDE. probably through the food chain, and
probably not mainly from the Aedes aegypti spray
program.

In the Miami area, a domestic turkey showed a brain
level of DDT -)- DDD high enough to seriously en-
danger the bird, possibly to be the cause of its death;
and one healthy loggerhead shrike showed heavy ex-
posure to DDE, probably through the food chain.

Seven other sick birds and five other dead birds col-
lected in the treated areas had brain levels of pesticides
that were so low as to rule out DDT as being the cause
of their illness or death.

Brain levels of DDT and its metabolites were not sig-
nificantly different between house sparrow nestlings
from treated zones and those from untreated zones in
the Miami area.

The great decrease in complaints and reports of wild-
life damage was probably correlated with the change in
operational spray methods employed; i.e., to more
selective applications and reduced rate of dosage with
insecticide.

The data herein presented are limited in sample size.
Specimens were collected over a span of only 4 months
and within only a small portion of the total area covered
by the spray program. The data are presented solely as
an indication of the effects of the Aedes aegypti Eradi-
cation Program on wildlife, with the recommendation
that studies of this type continue for the duration of
the program.

The chemic.ll names of compounds mentioned in this paper are:
DDT 1.1. 1 -trichIoro-2.2-bis( p-chlorophenyl ) ethane

DDD l.l-dlchloro-2.2-bis(p-chlorophenyl) ethane

DDE 1 . 1 -dichloro-2,2-bis( p-chlorophenyl ) ethylene

BHC 1,2.-^4. 5. 6-hexachlorocyclohexane. mixed isomers

Dieldrin not less than 85^- of 1.2..1.4.10.1()-hexachloro-

6, 7-epoxy-l, 4.4a. 5. 6,7,8, 8a-octahydro-l, 4, fnrfo-

c.To-5.8-dimethanonaphthalene
Hepiachlor epoxide I.4,5.6.7.8,8-heptachloro-2.3-epoxy-

.'^a.4.7.7a-tetrahydro-4.7-methanoindan.

A cknowledgment

The authors are indebted to the Patuxent Wildlife Re-
search Center of the U. S. Fish and Wildlife for assist-
ance in directing the study, especially to Mr. William
H. Stickel and Dr. Lucille F. Stickel, who provided
timely direction and assistance in the interpretation of
findings.

LITERATURE CITED

(I) Stickel. Lucille F.. William IL Stickel. and R. Christcn-
xen. 1966. Residues of DDT in brains and bodies of
birds that died on dosage and in survivors. Science
15U3717):1549-1551.

Pesticides Monitoring Journal

Pesticides in Hatchery Trout — Differences Between
Species and Residue Levels Occurring in Commercial Fish Food

H. Cole", A. Bradford-', D. Barry', P. Baiimgarner', and D. E. H. Frear'

ABSTRACT

Samples of commercial fish food from four manufaclurers
and trout of three species, brook, brown, and rainbow, were
analyzed for persistent chlorinated pesticides. The trout
were in the 8- to 9-inch size range at the time of analysis.
Small quantities of heptachlor, heplachlor epo.xide, dicldrin,
DDE, ODD (TDE), o,p'-DDT, and p.p'-DDT were found
in the fish food. One source contained all of the pesticides
except DDD. The trout were analyzed on the basis of
chloroform-methanol extractable lipids from the whole fish.
The rainbow trout with one exception contained all seven
pesticides. The rainbow trout contained greater quantities
of all pesticides than the brook or brown trout. The brown
trout contained significantly more heplachlor. heptachlor
epoxide, dieldrin, DDE, and DDD than the brook trout.

Introduction

VARIOUS investigations have demonstrated the pres-
ence of trace quantities of DDT and other persistent
chlorinated pesticides in feed stuffs including grains,
meat scraps, alfalfa meals, and fish oils. Many of these
ingredients are normally included in the manufacture
of commercial fish foods used in the production of
trout. In the investigation reported here an attempt was
made to determine the levels of certain persistent
chlorinated pesticides in commercial fish food and the
levels of pesticides occurring in trout being fed this
food at the Pennsylvania Fish Commission's Benner
Spring Research Station.

Sampling Methods

The fish meal samples were collected from commercial
packages of pellets. A 2-lb composite sample of pellets
was collected from lots of each of four different manu-
facturers. Three species, rainbow (Salmo gairdneri).
brown (Salmo trutta), and brook trout (Salvelinus fon-
tinalis) were collected from hatchery pools. Seven fish
of each species ranging in size from 8 to 9 inches were
used for analysis. All three species had been fed the
same brand of food (listed in Table 1 as source No. 1 )

' Pesticide Research Laboratories. Departments of Plant Pathology
and Entomology, The Pennsylvania State University, University
Park, Pennsylvania 16802. (Authorized for publication as paper
No. .1265 in the Journal Series of the Agricultural Experiment Sta-
tion on May 24, 1967.)

- Chief Fishery Pathologist, Pennsylvania Fish Commission, Benner
Spring Research Station, Bellefonte, Pennsylvania 16823.

in the same manner throughout their growth from the
fingerling stage until collected for analysis.

Analytical Methods

FISH FOOD

The samples of food consisting of 2 -lb of pellets were
ground in a Wiley Mill. A 100-g subsample of the
resulting meal was extracted for 16 hours in a large
Soxhiet apparatus with chloroform-methanol (2:1.
v/v). The extract was filtered with suction through a
Biichner funnel and placed in a flask equipped with a
Synder column. This was heated on a steam bath to
evaporate the chloroform. Two hundred ml of n-hexane
were added to the methanol and after thorough shaking
the mixture was washed three times with water to re-
move the methanol. The n-hexane extract was then
passed through an anhydrous sodium sulfate column to
remove the water. The extract was then passed through
a column of alumina, Celite, and Nuchar activated
carbon (2:1:1). This removed any pigments and other
interfering substances. The purified extract was con-
centrated in a Kuderna-Danish evaporator to a volume
of 2 ml and an aliquot injected into the gas chromato-
graph.

FISH

Each fish was weighed and then macerated in a Waring
Blendor with 300 ml of chloroform-methanol (2:1);
approximately 100 g of anhydrous sodium sulfate were
added during the blending process. The liquid was
decanted, and the blending repeated with another 300-mI
portion of chloroform-methanol. The extracts and
slurry were combined and filtered with suction through
filter paper in a Biichner funnel. The filtered extract
was then placed in a flask equipped with a Snyder
column. This was placed on the steam bath and the
chloroform removed. One hundred ml of n-hexane
were added, the liquid transferred to a separatory fun-
nel and washed three times with 200-ml portions of
water to remove the methanol. The washed extract was
filtered through anhydrous sodium sulfate, then
evaporated to a small volume in a flask on the steam
bath and then to dryness with a stream of air at room
temperature.

Vol. 1, No. 2, September 1967

35

At this point the residue consisted of lipid material in
a semisolid state. Tsvo g of this were weighed into a
small separalory funnel and dissolved in 25 ml of
petroleum elhcr. This solution was extracted by shaking
for 1 minute with 25 ml of acetonitrile saturated with
petroleum ether: the acetonitrile layer was drawn off and
the lipid solution re-extracted with three additional
25-ml portions of acetonitrile saturated with petroleum
ether. The combined acetonitrile extracts were evapo-
rated to a small volume and taken up in n-hexane. This
was then evaporated in a Kuderna-Danish evaporator
to exactly 2 ml and an aliquot injected into the gas
chromalograph.

Instrumental Procedure
All analyses were made on a Research Specialties Gas
Chromalograph Model 600, equipped with a 6-foot
glass column packed with Gas Chrom Q impregnated
with 59c DC-200. An electron capture detector was
used in all studies reported in this paper. The column
temperature was maintained at 210 C, the detector at
270 C, with a nitrogen flow of 60 ml per minute.
Samples of standard solutions were run periodically
to check on recovery. All results were calculated on
the basis of ppm of pesticide in the 2-g aliquot of lipid
material. Thus the results are on a "fat" basis derived
from the chloroform-menthanol extractable lipids. Con-
sidering the size of sample and analytical method, the
minimum level of detectability was considered to be
0.002 ppm. Residue traces less than 0.002 ppm were
reported as NR (no residue).

The pesticides included for analysis were heptachlor,
heptachlor epoxide, dieldrin, DDE, DDD (TDE),
o.p'-DDT. and p.p'-DDT. The identities of questionable
compounds were confirmed with a QF-1 column and
by thin layer chromatography.

Results

Tables I and 2 summarize the findings from analysis of
the fish meal and trout samples. The No. 1 food sample
was of the same brand that composed the diet of the
trout selected for analysis. It contained all the pesticides
included for analysis except DDD.

All three species of trout contained pesticides. All rain-
bow trout samples contained with a few exceptions
every pesticide included in the analysis. The rainbow
trout contained greater quantiiios of all pesticides than
the brook and brown trout I lu hrook and brown trout

The chemical names of compuundii mcnlioned in this paper are:
DDT 1 , 1 . l-lrichloro-2.2-bis(p^:hlorophenyl ) ethane

DDD l.l-<)ichloro-2.2-bis(p.chlorophcnyI)cthane

DDE 1 . 1 -dichloro-2,2-bis( p-chlorophcnyl ) ethylene

BHC 1,2,-1. 4. 5.6-hexachIorocyclohcxane, mixed isomers

Dieldrin not less than 85% of 1.2,3.4. lO.lO-hexachloro-

6-7-epoxy-l,4.4a,5.6.7.8.8a-octahydro-l,4-0Hdo-

fjrr»-5.8-dimethanonaphlhalenc
Heptachlor 1.4.5,6.7.8.K-hcpiachloro-3a.4,7.7a-letraliydro-

4.7-mcthanoindenc
Heptachlor epoxide l,4.5.6.7,8,8-heplachloro-2.3-epoxy-.1a,4,7,7a-

tetrahydro,4,7-me(hanoindan

did not contain any of the p.p'-DDT isomer. Statistical
treatment of the results by analysis of variance and
studcntized range test indicated that the rainbow trout
contained significantly more (0.05 confidence level)
heptachlor, heptachlor epoxide, dieldrin, DDE, DDD,
o.p'-DDT. and p.p'-DDT than either the brook or brown
trout, and that the brown trout contained significantly
more heptachlor, heptachlor epoxide, dieldrin, DDE,
and DDD than the brook trout. The lipids extractable
in chloroform-methanol represented about 2.5% of the
fresh weight of the fish. Thus, an approximate fresh
weight pesticide content may be obtained by dividing
the results in Table 2 by a factor of 40.

Discussion

Previous research has shown that fish vary in their
tolerance to pesticides as evidenced by widely different
LCr,,, values from species to species (4). It has also been
shown that pesticide resistance is present in certain
strains of mosquitofish (Gamhiisia affinis) golden shiners
(Notemiqonus crysoleucas), green sunfish (Lepomis
cyonelliis). bluegills (Lepomis macrochirus). and yellow
bullhead (Iclaliinis nalalis) (1-3).

It also has been shown that individual lots of fish from
different sources vary markedly in their LC.^,, values.
For example Marking (4) found that with p.p'-DDT,
rainbow trout lots varied from LCr,,, ppb values of 2.4
to 17 and brook trout from 1.8 to 20. In the present
study it has been shown that when three species of
trout are fed the same diet throughout a prolonged
period, the whole body accumulation of certain pesti-
cides varies considerably with the species. All trout in
the pools from which the samples were selected ap-
peared to be in normal health and all trout in the
hatchery including breeding stock were being fed the
No. 1 brand of fish food. The hatchery at the Research
Station has indicated no reproductive problems to date.
Analyses of water from sources entering the hatchery
have failed to show the presence of pesticides in waters
entering the hatchery.

It is also of interest that while pesticide residue toler-
ances for fish have not been established, the levels found
in rainbow trout were above the levels accepted by the
FDA for beef fat.

It is not known whether the differences between species
represent differences in uptake, excretion, or degradation
of pesticides. It is also not known how much the diet of
the fish may influence uptake and accumulation. How-
ever, it appears in the case of DDT at least, degradation
abilities between species may vary since the brook and
brown trout under the conditions in the study did not
contain any p.p'-DDT isomer.

At present studies are underway to determine if various
genetic lines within species with uniformity for other
characters will exhibit uniform differences in pesticide
accumulation when fed similar diets.

36

Pesticides Monitoring Journal

LITERATURE CITED

(/) Boyd, C. E. and E. E. Ferguson. 1964. Susceptibility

and resistance of mosquitofish to several insecticides.

J. Econ. Entomol. 57:430-431.
(2) Ferguson. D. E. and C. R. Bingham. 1964. Endrin

resistance in the yellow bullhead, Iclalurus natalis.

Trans. Amer. Fish Soc. 95:325-326.

(3) Ferguson, D. £., D. D. CuUey, W. D. Cotton, and
R. P. Dodds, 1964. Resistance to chlorinated hydro-
carbon insecticides in three species of fresh water fish.
Bioscience 14(1 1):43-44.

(4) Marking, L. L. 1966. Evaluation of p.p'-DDT as a ref-
erence toxicant in bioassays. USDI Resource Publica-
tion No. 14:1-10.

TABLE 1 . — Pesticides in commercial fish food

PPM

Food

Heptachlor

SOURCE

Heptachlor

EPOXIDE

DlELDRIN

DDE

ODD

o,p'-DDT

p,p'-DDT

1

0.073

0.014

0.060

0.096

NR

0.305

0.087

2

0.198

NR

0.085

0.315

0.189

NR

NR

3

0.101

0.211

NR

NR

NR

NR

NR

4

0.031

NR

NR

0.016

NR

NR

NR

Notes: Each source represents the product of a different manufacturer. Samples were collected on 3/31/66.
NR = Less than 0.002 ppm.

TABLE 2. — Pesticides in hatchery trout of three species from Benner Spring Hatchery'

PPM

2

Sample

Heptachlor

No.

Species

Heptachlor

epoxide

DlELDRIN

DDE

DDT

o,p'-DDT

p,p'-DDT

1

Rainbow

0.5

1.3

0.8

1.3

3.2

1.1

4.0

2

Rainbow

0.5

1.0

0.7

1.3

1.8

1.0

2.7

3

Rainbow

0.4

0.9

0.6

1.0

2.1

0.9

2.7

4

Rainbow

0.5

1.3

0.8

1.3

3.2

1.2

4.0

5

Rainbow

0.7

1.4

0.8

1.4

2.2

0.8

2.8

6

Rainbow

0.4

1.2

0.9

1.3

3.3

0.9

3.5

7

Rainbow

2.8

7.5

0.5

NR

4.4

NR

5.8

Species mean

0.8

2.1

0.7

1.1

2.9

0.8

3.6

8

Brown

0.03

NR

0.05

0.81

1.3

0.02

NR

9

Brown

NR

NR

0.006

0.34

0.28

NR

NR

!0

Brown

0.06

NR

NR

0.82

1.3

NR

NR

11

Brown

0.02

NR

NR

0.68

0.81

NR

NR

12

Brown

0.08

0.09

NR

0.12

0.19

NR

NR

13

Brown

0.003

0.13

O.008

0.20

0.40

NR

NR

14

Brown

0.006

0.14

0.03

0.19

0.41

NR

NR

Species mean

0.03

0.05

0.01

0.45

0.67

0.003

15

Brook

0.48

NR

0.02

0.07

0.68

NR

NR

16

Brook

NR

NR

NR

0.45

NR

NR

NR

17

Brook

NR

NR

NR

0.15

0.24

NR

NR

18

Brook

NR

NR

NR

NR

NR

0.002

NR

19

Brook

NR

NR

0.10

0.15

0.70

NR

NR

20

Brook

0.009

NR

NR

NR

0.003

NR

NR

21

Brook

NR

NR

0.003

0.05

0.34

NR

NR

Species mean

0.007

0.002

0.12

0.28

I Fish selected at random from hatchery pool of each species at Benner Spring. All lish in 8- to 9-inch size category. Each sample number

represents a single fish.
- Based on amount of pesticide in the lipids exiractable by chloroform-methanol.
Note: NR = Less than 0.002 ppm.

Vol. 1, No. 2, September 1967

37

PESTICIDES IN WATER

Pesticides in Selected Western Streams — A Contribution
to the National Program'

E. Brown and Y. A. Nishioka

ABSTRACT

Since October 1965. samples of a waler-suspcndcd sediinenl
mixliire from II streams in the western United SlcUcs liave
been analyzed montlily for 12 pesticides. The compounds
determined include the insecticides aldrin. DDD, DDE,
DDT, dieldrin, endrin, heptachlor, heptachlor epoxide, and
lindane; and the herbicides 2,4-D: 2.4.5-T: and silvex. No
herbicide was found at any station during the first year of
the sampling program. All insecticides were found at one
lime or another, but not at all stations. Tlie amounts ob-
served were quite snuill, ranging from le.^s than 5 parts per
trillion of lindane to 110 parts per trillion of DDT.

Inirodiictioii

IN the fall of 1965, the U. S. Geological Survey initiated
a limited program of pesticide monitoring on 1 1 streams
in the western United States, selected from the Survey's
program of water-quality studies of irrigation-network
sites. Purpose of the program was to determine the
extent and magnitude of pesticide contamination. To
accomplish this, the streams were analyzed initially for
nine of the more commonly used insecticides; analysis
for herbicides was begun later in the program. Insecti-
cides chosen for analysis included aldrin, DDD, DDE,
DDT, dieldrin. endrin, heptachlor. heptachlor epoxide,
and lindane. The herbicides consisted of 2,4-D; 2,4,5-T;
anil silvex.

Pesticides selected for analysis were chosen mainly from
the primary list of pesticide compounds established in
March 1964 by the Subcommittee on Monitoring, Fed-
eral fommittee on Pest Control.

Data Collection Sites

In selecting the actual sampling sites, consideration was
given to the following criteria: (I) each station should
be a designated or operating U. S. Geological Survey
irrigation-network location; (2) where practical, each
station should be considered as one of the sites for the
minimum national pesticide monitoring program recom-

' Water Resources Division. U. S. Geological Survey, Sncr.imento.
California 95KI4. (Puhlication auihorized by Ihc Director. U. S.
GcoloKical Survey).

mended by the Federal Committee on Pest Control; (3)
each site should be one at which other types of data are
being obtained; (4) no station should overlap the
activities of other agencies. It was felt that irrigation-
network stations were highly desirable because: (1)
several years of record of inorganic water quality and
stream discharge are available; and (2) these stations
represent mainly agricultural areas where the proba-
bility of observing pesticide residues would be greater.

Stations selected for sampling are listed in Table 1 and
their location shown in Fig. I. Complete hydrologic

TABLE I. — Pesticide monitoring stations in western
United Stales

Irri-

Geological

gation

Survey

Inorganic

NETWORK

STATION

analysis

NO.l

IDENT. NO.

Stream and location

STARTED

4

6-8070

Missouri River at Nebraska
City. Neb.

1-4-51

24

7-1305

Arl<ansas River below John
Martin Reservoir, Colo.

1-10-51

27

-2505

Arkansas River at Van
Buren, Ark.

10-1-45

37

8-1140

Brazos River at Richmond,
Tex.

9-1-45

40

-1620

Colorado River at Wharton,
Tex.

4-11-44

52

^625

Rio Grande below Anzalduas
Dam, Texas

1944

63

9-5255

Colorado River (Yuma Main
Canal) below Colorado
River Siphon, at Yuma,
Ariz.

1042

86a

11-4255

Sacramento River at Verona,
Calif.

94

12-5105

Yakima River at Kiona,
Wash.

12-30-52

97

13-1545

Snake River at King Hill,
Idaho

3-27-51

102

14-1057

Columbia River near The

12-1-50

Dalles, Ore.

Number refers to list of irrigation-quality network stations (U. S.
Geological Survey, 1954, p. 3).

38

Pesticides Monitoring Journal

data for these stations are published in annual reports
of the U. S. Geological Survey entitled "Quality of sur-
face waters for irrigation, western United States." These
reports include inorganic water-quality data, drainage
area and stream discharge figures, as well as an indica-
tion of the period of record available. The first report
was issued in 1954 as U. S. Geological Survey Water-
Supply Paper 1264 (4) and covers the period October
1, 1950 through September 30, 1951. The latest report
in this series was released in 1966 as Water-Supply
Paper 1946 (5) and contains data for the period October
1, 196! through September 30, 1962.

Sampling Procedures

At the beginning of the program, samples were collected
monthly in I -gallon Pyrex bottles by personnel of the

U. S. Geological Survey, with the exception of the
station below Anzalduas Dam, Texas. In this case,
samples were provided by personnel of the International
Boundary and Water Commission. United States and
Mexico, United States Section. The bottles were tightly
stoppered with rubber stoppers, wrapped in aluminum
foil, and promptly shipped in wooden boxes to the
laboratory for analysis. The size and weight of the bot-
tle and container required shipment by rail, so that in
most cases 2 to 3 weeks elapsed between collection and
analysis. In addition, containers were broken and
samples lost in transit.

As soon as analytical methods improved to the point
that a smaller sample could be used without sacrifice of
accuracy, the duo-pak container was put into use. This
container (/ ) is lightweight, small, and sufficiently sturdy

FIGURE 1. — Pesliciile stations in western United States

EXPLANATION

Boundary line of principal drainage basin

Pesticide station in operation, 1966

Number refers to station selected from "Quality of Surface Waters for Irrigation,
Western United States— 1951." U.S. Geological Survey Water Paper 1264

Vol. 1, No. 2, September 1967

39

FIGURE 2. — Class sample conuiiner, screw-cap (Teflon
lined}, and expanded polystyrene proieclive case

to permit shipment by air express. No breakage in
transit has been observed during 6 months of use. The
collection unit— consisting of plastic foam case, bottle,
cap. and teflon capliner — is shown in Fig. 2.

Two bottles were collected at each station, one being
used for insecticide analysis and the other for herbicides.

It was originally intended that a depth-integrated sample
be collected to most nearly represent the average water-
quality condition at the time of sampling. Small-mouthed
gallon jugs provide a reasonable approximation of this
type of sample; wide-mouth quart jars however, fill
almost instantly when lowered into the water. The type
of sample obtained is not representative of the vertical
section, but only of the upper-most water layer.

At the present time, a study is underway to modify
existing sediment-sampling equipment to provide both
depth and point-integrated samples,

A nalyiical Procedures
Because the total insecticide or herbicide concentration
was desired, each sample was analyzed as received,
with no attempt to separate the water and sediment for
individual analysis. Chlorinated pesticides were analyzed
by the method described by Lamiw et al. (3) which is
summarized below: one liter sample of water was ex-
tracted three limes with equal volumes of hexane to a
total of 75 ml of hexane. Hexane was dried over an-
hydrous Na,SO, and concentrated to 5 ml in a Kuderna-
Danish evaporator. Of the concentrated hexane, 5 ^1

40

were injected into the gas chromatograph. In all cases,
injections were made into two dilTerent chromatographic
columns to effect separation.

Recovery data reported by Lamar el al. (3) range from
about 80"^ to I 159f . No adjustments were made to the
data reported in this paper, because many of the values
are so near the lower limits of sensitivity that they are
rounded off to the nearest 5 ppt.

Herbicide analysis was conducted according to a pro-
cedure developed by Goerlitz and Lamar (2). using
boron irifluoride methanol reagent for esterification. As
in the analysis of insecticides, a liter sample was used,
with the final volume of extractant being reduced to 5
ml prior to injection into the gas chromatograph. Re-
covery data reported by Goerlitz and Lamar (2) ranged
from about 75% to 120%. No adjustments were made
in the data reported for herbicides.

Operating conditions for the chromatographic procedure
were as follows:

Instrument:

Columns;

Aerograph Hy-FI Model 600-D,
with a Wilkens Model 328
Isothermal temperature con-
troller

Oven temperature:

Detector:

Carrier gas:

Injection volume;

Vs" X 5' pyrex glass, packed
with 60/80 mesh Gas-Chrom
Q-coated with 5% DC 200.
Vs" X 5' pyrex glass, packed
with 60/80 mesh Gas-Chrom
Q-coated with 5% QF-L

187C

Electron-capture, concentric tube
design, D.C. mode

Nitrogen at 40/min

5/il

Using these procedures, accurate analysis of most water
is routinely practical if it contains the minimum concen-
tration of pesticides indicated in Table 2. Amounts less
than that can be detected, but are not considered ac-
curately measurable unless a larger sample volume is
taken for analysis, or the extractant volume is reduced
to less than 5 ml. For example, water containing as little
as 10 nanograms per liter of 2,4-D may be analyzed if
the final extraction volume is reduced to 0.5 ml instead
of 5.0 ml. Not all extracts, however, can be reduced to
such a low volume without an accompanying buildup of
excessive interferences. The extensive cleanup necessary
to remove such interference is not alwavs practical in
routine analysis.

Pesticides Monitoring Journal

TABLE 2. — Minimum measurable concentrations of pesticides in water

Parts pi:r

Parts per

TRltLlON

TRILLION

Pesticides

( NANOGRAMS

Pesticides

(NANOGRAMS

PER LtTER)

PER LITER)

ALDRIN

5

HEPTACHLOR

5

not less Ihan <)5'7 of 1,2.3,4,10,10-hexachloro-

1,4,5 ,6.7.8, 8-heptachloro-3a,4,7,7a-tetrahydro-4.7-

1,4,4a, 5.8. Sa-hexahydro-l,4-endo-MO-5,8-

methanoindene

dimethanonaphlhalene

DDD

l,l-dichloro-2,2-bis(;i-chloriiphenyI)elhane

5

HEPTACHLOR EPOXIDE

5

1,4,5,6,7, 8, 8-heptachloro-2.3-epoxy-3a,4,7,7a-
tetrahydro-4,7-methanoindan

DDE

5

I. l-dichloro-2,2-bis(p-chlorophenyl) ethylene

LINDANE

1,2.3,4,5,6-hexachlorocyclohexane, 99% or rnoic

5

DDT

10

gamma isomer

1,1,1 -trichloro-2,2-bis( p-chloropheny 1 ) ethane

2,4-D

100

DIELDRIN

not less than 85'S of l,2,3.4,10,10-hexachloro-6.7-

5

2,4-dichlorophenoxyacetic acid

epoxy- 1 ,4,4a,5,6,7,8,8a-octahydro-l ,4-e«do-e.vc'-

2,4, 5-T

5

5,8-djmethanonaphthaIene

2,4,5-trichlorophenoxyacetic acid

ENDRIN

5

l,2.3,4,10,10-hexachloro-6,l-epoxy-l,4,4a, 5.6.7,8, 8a-

SILVEX

5

Octahydro-l,4-piido-cndo-5,8-dimethanonaphthalene

2-(2,4,5-trichlorophenoxy Ipropionic acid

Discussion of Results

Pesticide analyses of samples of a water-suspended
sediment mixture obtained during the first year of this
study are arranged in downstream order and presented
in Table 3. Data in Table 3 have been summarized and
presented in Table 4 to indicate frequently occurring
pesticides, and the stations at which they occurred.
Based on data obtained, the following observations can
be made:

1. No herbicide was found at any time at any station.
The absence of herbicides from any of the samples
analyzed may be due at least in part to their suscepti-
bility to degradation. Recent studies by Goerlitz and
Lamar (2) on the stability of herbicides added -to water
samples, indicate that 2,4-D may be degraded rapidly,
especially in samples obtained from areas repeatedly
treated with 2,4-D.

2. All insecticides were found at one time or another,
but not at all stations.

3. The insecticide concentrations found were very
small, amounting to 5 ppt in slightly more than 50%
of all positive results.

4. Although no definite pattern could be noted in
pesticide occurrence, positive results were most fre-
quently found in February, March, April, and May.

5. The most frequently found insecticide was lindane,
which occurred 46 times out of a total of 165 positive
results. The most infrequently occurring insecticide was
aldrin, which was observed only four times at all stations
during the year.

6. On a geographical basis, most frequent occurrence
of pesticides was at the Rio Grande Station below

Anzalduas Dam, Texas. Thirty-two positive results
were obtained at this station, or about 20% of the total.
The least number of positive results were noted at the
Snake and Columbia River stations, each of which
recorded only seven pesticide occurrences.

7. No relationship can be noted between pesticide oc-
currence and the various factors in pesticide application
since the latter parameters cannot be ascertained in
most areas.

8. No evaluation of other forms of pesticide movement,
such as might be involved in sediment transport, can be
made on the basis of the data obtained. Data presented
will provide a basis for efforts into other areas of pesti-
cide relationships, such as mode transport, time of
travel, and improved sampling techniques.

LITERATURE CITED

(I) Breidenbacli, A. W.. J. J. Liclitenberg. C. F. Henke,
D. J. Smith. J. W. Eichelberger. Jr., and H. Stierli.
1964. The identification and measurement of chlorinated
hydrocarbon pesticides in surface waters. U. S. Dep.
of Health, Educ, and Welfare, Public Health Serv., Pub.
No. 1241, p. 11.

{2) Goerlitz, D. F., and W . L. Lamar. Determination of
phenoxy acid herbicides in water by electron-capture
and microcoulomelric gas chromatography. U. S. Geol.
Surv. Water-Supply Paper 18I7-C. (in press).

(3) Lamar, W. L., D. F. Goerlitz. and L. M. Law. 1965.
Identification and measurement of chlorinated organic
pesticides in water by electron-capture gas chromatog-
raphy. U. S. Geol. Surv. Water-Supply Paper 1817-B.

(4 ) U. S. Geological Survey. 1954. Quality of surface waters
for irrigation, western United States, 1951. U. S. Geol.
Surv. Water-Supply Paper 1264. 153 p.

(5) U. S. Geological Survey. 1966. Quality of surface waters
for irrigation, western United States. 1962. U. S. Geol.
Surv. Water-Supply Paper 1946, 143 p.

Vol. 1, No. 2, September 1967

41

TABLE 3. — Pesticide content of selected streams in western United States
nd — not determined; — — not present

Parts per trillion (nanograms per liter.

rounded to nearest 5 PPT)

DC

M

o

2"

Instan-

z

ul

taneous

2

g

z

o

U X

z

H

X

Dis-

cc

Q

u

H

u

PC

H

£ 2

o

Q

*n

Date

Time

THARGE
(CFS)

<

a
a

Q
Q

Q

Q

z

X

aw

X

z

3

fi .

[75

IRRIG. NETWORK NO. 4— USGS NO. 6-8070 MISSOURI RIVER AT NEBRASKA

10/18/65

10:30A

38.700

—

—

—

—

10

—

—

—

—

nd

nd

nd

11/2.1/65

lO.OOA

39,700

-

-

-

—

-

—

-

-

5

nd

nd

nd

12/08/65

1I:30A

25,900

-

—

—

-

—

-

-

-

-

nd

nd

nd

01/20/66

2:30P

20,400

-

-

—

-

-

-

-

-

-

nd

nd

nd

02/09/66

2:30?

47,900

—

—

—

—

5

—

-

-

-

nd

nd

nd

0.VO.V66

10:45A

32,600

-

-

-

—

15

—

-

-

5

nd

nd

nd

04/04/66

I0:45A

40,000

-

-

-

—

5

—

-

~

5

-

—

—

05/17/66

1:30?

35,600

5

-

50

15

35

-

5

5

-

—

—

06/15/66

10:00A

37,200

—

-

—

—

—

-

-

—

-

—

—

07/07/66

10:45A

35,900

—

—

—

—

—

—

—

5

—

-

—

—

08/04/66

10:00A

37,200

-

—

-

-

—

—

5

-

-

-

—

—

09/16/66

10:30A

36,000

—

—

—

45

—

—

—

—

—

—

—

—

IRRIG. NET. NO. 24— USGS NO. 7-1305 ARKANSAS RIVER BELOW JOHN MARTIN RESERVOIR, COLO.

10/16/65

7:45A

272

—

—

5

—

5

—

5

5

5

nd

nd

nd

11/26/65

3:00?

297

—

—

—

—

—

—

—

—

10

nd

nd

nd

12/21/65

4:00?

331

—

—

—

—

—

—

—

5

nd

nd

nd

01/20/66

4:00?

297

—

—

—

—

—

—

—

—

—

nd

nd

nd

02/19/66

8: 30 A

448

—

—

—

—

5

—

—

5

10

nd

nd

nd

03/22/66

8:40A

87

—

—

—

—

—

—

—

—

5

nd

nd

nd

04/18/66

2:40?

774

—

—

—

—

5

—

—

—

10

—

—

—

05/16/66

3.20?

1,030

—

10

.s

75

—

—

in

—

10

—

—

—

06/13/66

3:15?

870

—

—

—

—

—

—

—

—

—

—

—

07/25/66

4:20?

978

—

—

15

—

—

—

5

5

—

—

—

—

08/23/66

9:45A

283

—

—

—

—

—

—

—

—

—

—

—

—

09/19/66

3:50?

506

—

—

—

—

—

—

-

—

5

—

—

—

IRRIG. NET. NO. 27— USGS NO. 7-2505 ARKANSAS RIVER AT VAN BUREN. ARK.

10/21/65

3:40?

12,800

—

20

.

nd

nd

nd

11/22/65

I1:00A

3,960

—

—

—

—

—

—

—

—

—

nd

nd

nd

12/30/65

'' o.SA

7,200

—

—

—

—

5

—

5

—

5

nd

nd

nd

01/11/66

12:20?

6,250

—

—

—

—

—

—

—

—

—

nd

nd

nd

02/07/66

12:30?

5,070

—

—

—

—

5

—

—

—

5

nd

nd

nd

03/09/66

9:00A

11,000

-

5

5

—

—

—

—

5

in

nd

nd

nd

04/06/66

8:50A

5,520

—

—

—

—

5

_

—

—

5

—

05/10/66

1:05?

16,700

—

—

—

—

—

—

—

—

5

06/16/66

12:45?

23,100

5

-

-

-

—

—

5

-

—

—

—

—

42

Pesticides Monitoring Journal

TABLE 3.— Pesticide conlent of selected streams in western United Slates— Conlinued
nd = not determined; — r^ not present

Parts per trillion (nanograms per liter

ROUNDED TO NEAREST 5 PPT

)

Instan-

z

S^fl

taneous

z

2

u

U X

Dis-
charge

Q
-J

D
O

Q

s

a

OS

o

z

a.
u

H a.

z

Q

UJ

>

Date

Time

(CFS)

<

Q

Q

"

Q

w

I

X

.J

<N

ri

c/)

IRRIG. NET. NO. 27— USGS NO. 7-2505 ARKANSAS RIVER AT VAN BUREN, ARK.— Continued

20A
30A
30A

3,280
24,400
13,400

10

70
110

10
10

IRRIG. NET NO. 37— USGS NO. 8-1140 BRAZOS RIVER AT RICHMOND, TEX.

Oct. 1965

No sample

Nov. 1965

No sample

Dec. 1965

No sample

02/01/66

12.00?

5,620

—

5

—

—

10

—

—

5

5

nd

nd

nd

Mar. 1966

No sample

04/12/66

12:50?

1,930

—

—

5

—

—

—

—

5

5

—

—

—

05/19/66

2:00?

22,200

-

10

—

55

15

—

—

—

5

—

—

—

06/22/66

1I:30A
3:15?
11:00A

2,900
1,570
3,460

—

—

—

45

10

—

15

—

—

—

—

—

08/30/66

-

-

10

105

—

—

—

—

—

IRRIG. NET. NO. 40— USGS NO. 8-1620 COLORADO RIVER AT WHARTON, TEX.

10/22/65

11:30A

1,600

—

—

—

25

5

5

5

5

—

nd

nd

nd

Nov. 1965

No sample

Dec. 1965

No sample

01/07/66

9: 00 A

1,400

—

—

—

—

10

—

—

5

10

nd

nd

nd

02/02/66

3:15?

897

—

—

—

—

10

—

—

—

5

nd

nd

nd

Mar. 1966

No sample

04/13/66

8:30A

1,630

—

—

15

70

—

—

-

-

20

-

—

—

05/24/66

5:30?

5,490

—

—

5

—

—

—

-

—

—

—

—

—

06/27/66

3:00?

1,320

—

—

—

—

—

—

—

—

—

—

—

07/28/66

5:30?

1,380

—

—

—

—

—

—

-

-

—

—

-

—

Aug. 1966

No sample

09/01/66

9:30A

483

-

—

—

—

—

—

—

—

—

—

—

—

IRRIG. NET. NO. 52— USGS NO. 8^625 RIO GRANDE BELOW ANZALDUAS DAM, TEX.

10/15/65

9:10A

500

5

5

10

—

10

5

10

nd

nd

nd

11/26/65

8:30A

300

—

10

—

—

15

—

—

—

5

nd

nd

nd

12/15/65

1:45?

260

—

—

—

—

15

—

15

-

10

nd

nd

nd

01/14/66

8:30A

300

—

—

—

—

10

—

5

—

5

nd

nd

nd

02/16/66

9: 30 A

850

—

—

5

—

—

10

—

—

5

nd

nd

nd

03/14/66

9:10A

920

-

—

—

—

—

—

-

10

10

nd

nd

nd

Vol. 1, No. 2, September 1967

43

TABLE 3. — Pcslicitic conlciit of selected streams in western United Stales — Continued
nd = not determined; — = not present

Parts pek trillion (nanograms per liter

ROUNDED TO NEAREST 5 PPT)

tA

tc

o

9 UJ

INSHN-

2

i

^e

u

MNIOUS

7

g

z

a
z

<

<g

z

H

X

Dis-

(HARUb

Q
<

a
a

Q

UJ

D

n

H
Q
Q

Q

X

a
z

0
4,

>

Dais

Time

(CFS)

IRRIC. NET. NO. 52— USGS NO. 8-4625 RIO GRANDE BELOW ANZALDUAS DAM, TEX.— Continued

04/15/66

9:. 10 A

300

—

—

—

—

—

—

—

—

—

—

—

—

05/17/66

9:30A

280

—

10

10

—

—

40

15

-

10

—

—

—

06/16/66

8:30A

2,500

-

-

—

-

—

25

-

—

-

—

—

—

07/12/66

8:00A

7,950

-

15

-

50

10

—

—

—

—

—

—

—

08/15/66

9:00A

900

-

—

10

—

—

—

-

—

5

—

—

—

09/16/66

I:OOP

12.360

-

10

—

—

—

—

—

—

—

—

—

—

IRRIG. NET. NO. 63— USGS NO. 9-5255 COLORADO RIVER (YUMA MAIN CANAL) AT YUMA, ARIZ.

10/12/65

I0:30A

502

—

—

—

—

—

—

—

—

—

nd

nd

nd

12/01/65

2:00P

239

—

—

—

—

—

—

—

—

—

nd

nd

nd

01/05/66

IO:OOA

161

-

-

—

—

-

—

-

5

-

nd

nd

nd

02/01/66

IrOOP

239

—

—

—

—

—

—

—

—

—

nd

nd

nd

03/02/66

9: 30 A

616

—

—

—

70

5

—

—

—

5

nd

nd

nd

04/06/66

11:00A

216

—

—

—

—

—

—

—

—

5

—

—

—

05/03/66

I:30P

687

—

—

5

—

—

—

—

—

5

—

—

—

06/02/66

3:30P

661

—

—

10

—

—

15

—

90

—

—

—

—

07/05/66

9:30A

557

—

—

—

—

—

—

—

—

—

—

—

—

08/08/66

9:0OA

560

—

—

—

—

—

—

—

—

—

—

—

—

09/01/66

9:0OA

671

-

—

-

-

—

—

—

—

—

—

—

—

IRRIC. NET. NO. 86a— USGS NO. 11^255 SACRAMENTO RIVER AT VERONA, CALIFORNIA

Oct. 1965

No sample

11/29/65

lOiOOA

23,400

—

—

—

—

—

—

—

—

—

nd

nd

nd

12/17/65

10:05A

20,000

—

—

—

—

—

—

—

—

—

nd

nd

nd

01/06/66

10:OOA

39,600

—

—

—

—

—

—

5

—

nd

nd

nd

02/04/66

3: OOP

29,900

—

—

—

—

—

—

—

—

—

nd

nd

nd

03/03/66

in:2nA

l«,90fl

--

—

—

—

10

—

—

5

5

nd

nd

nd

04/01/66

2:00P

24,600

—

—

—

—

—

—

—

—

5

—

—

—

05/02/66

9:30A

11,700

—

—

—

—

5

—

—

—

5

—

—

06/03/66

1I:OOA

8,390

—

—

—

—

—

—

—

—

""

07/05/66

lOiOOA

8,940

—

—

—

—

—

—

—

08/16/66

9:0OA

10.700

—

10

—

—

—

Sept. 1966

No sample

IRIG. NET. NO. 94— USGS NO. 12-5105 YAKIMA RIVER AT KIONA, WASH.

10/11/65

1:15?

2,100

_

_

_

nd

nd

nd

Nov 1<w,^

Nn ■iampic

1.

1,710

—

—

—

—

—

—

-

-

-

lid

nd

nd

44

Pesticides Monitoring Journal

TABLE 3. — Pesticide content of selected streams in western United Slates — Continued
nd = not determined; — = not present

Parts per trillion (nanograms per liter,

rounded to nearest 5 PPT)

^

oe

o m

Instan-

z

^

&B

taneous

z

z

u

U X

z

H

Dis-

g

a

u:

H

CA

f-

5!e

2

Q

UJ

charge

u

o

Q

Q

S

z

LU

Sjw

z

-1

Date

Time

(CFS)

<

a

Q

Q

Q

w

X

S

-1

(N

crt

IRRIG. NET. NO. 94— USGS NO. 12-5105 YAKIMA RIVER AT KIONA, WASH.— Continued

Jan. 1966

No sample

02/02/66

4:45P

1,820

—

—

—

—

—

—

—

—

—

nd

nd

nd

03/04/66

3:45?

2,050

—

—

—

—

5

—

—

5

10

—

—

—

04/01/66

11:00A

6,390

-

-

10

—

—

—

—

10

—

—

—

05/09/66

11:55A

5,510

—

—

10

—

—

—

—

—

5

—

—

—

06/17/66

4:00P

1,680

—

5

—

—

—

—

—

—

—

—

—

—

07/25/66

11;15A

1,380

-

10

15

—

—

—

5

5

5

—

—

—

08/30/66

ll:0OA

1,820

—

—

—

65

—

—

—

—

—

—

—

—

Sep. 1966

No sample

IRRIG. NET. NO. 97— USGS NO. 13-1545 SNAKE RIVER AT KING HILL, IDAHO

10/17/65

2;00P

14,700

—

—

—

—

—

—

—

—

—

nd

nd

nd

11/29/65

2:55P

14,700

—

—

—

—

—

—

—

—

—

nd

nd

nd

Dec. 1965

No sample

01/03/66

2: OOP

15,900

—

—

—

—

—

—

—

5

—

nd

nd

nd

02/07/66

11:45A

14,400

—

—

—

—

—

25

5

—

—

nd

nd

nd

03/21/66

4:15P

15,000

—

—

—

—

—

—

—

—

—

nd

nd

nd

04/24/66

3:30P

11,400

—

—

—

—

5

—

—

—

5

—

—

May 1966

No sample

06/09/66

1:20

10,200

5

—

—

—

—

—

—

—

—

—

—

07/08/66

12:30P

9,750

—

—

—

60

—

—

—

—

—

~

—

—

08/23/66

10:30A

4,100

—

—

—

—

—

-

—

—

-

—

09/27/66

11:00A

6,420

—

—

—

—

—

—

—

—

—

—

IRRIG. NET. NO. 102— USGS NO. 14-1057 COLUMBIA RIVER AT DALLES, ORE.

10/27/65

5:30P

104,000

—

—

—

—

—

—

nd

nd

nd

11/22/65

9:30A

105,000

—

—

—

—

-

—

—

nd

nd

nd

12/27/65

I1:15A

101,000

—

—

—

—

10

—

-

-

—

nd

nd

nd

01/27/66

4:45P

125,000

—

—

—

—

—

—

—

5

-

nd

nd

nd

02/21/66

8: 45 A

102,000

—

—

—

—

—

—

—

—

5

nd

nd

nd

03/25/66

1:30P

120,000

—

—

—

—

—

—

—

-

5

nd

nd

nd

04/29/66

9: 00 A

171,000

5

—

—

—

—

—

-

-

5

-

—

—

05/23/66

4:00P

280,000

—

—

—

—

—

—

-

-

-

-

-

—

06/22/66

7:45A

319,000

—

—

—

—

—

—

-

-

—

—

—

—

07/14/66

1:30P

286,000

—

—

—

—

—

—

-

—

—

-

—

—

08/15/66

8:00A

145,000

—

—

—

—

—

—

-

-

20

—

—

—

09/19/66

8:15A

96,000

—

—

—

—

—

—

—

—

—

—

—

—

Vol. 1, No. 2, September 1967

45

TABLE 4. — Occurrence of insecticides

U

s

Arkansas River below
John Martin
Reservoir, Colo.

< CO
2 Z

<

X

* o'
> z

si

m

2 z

ii

H
a in

n

o<

<

o:

15

O 3

u

oU

If

>■

H

5"

01 u

is

u

.J

Aldrin

DDD

DDE

DDT

Dieldrin

Endrin

Heptachlor

Heptachlor epoxide

Lindane

1

1

3

1
3

3
3
S

1

2
2
2
5

2
1

5

2
2
3
3

1
2
3

2
2
3
I
1
2
3

5

4
1
5
3
4
2
8

->

1
1

2
3

1

2

2
3

2
3

1

1

2
4

1

1

1
1

4

4
13
18
14
29

7
14
20
46

Totals

16

22

20

16

14

32

10

8

13

7

7

165

46

Pesticides Monitoring Journal

Persistence and Movement of Parathion in Irrigation Waters^

C. W. Miller , W. E. Tomlinson. and R. L. Norgren

ABSTRACT

The occurrence, persistence, and movement of parathion
(0,0-dielliyl 0-p-nilroplienyl pliospliorolliioate) in cranberry
bog irrigation waters was investigated. Tlie chemical was
found to persist for 96 hours at concentrations known to be
toxic to certain aquatic organisms. Movement of the cliemi-
cal from the irrigation waters to an associated water system
was also demonstrated to occur; liowever, tlie degree of con-
centration and persistence was not as great as williin llic bog
area.

Introduction

APPLICATION of parathion to cranberry bogs, either
by heHcopter or through overhead sprinklers.is often
made with water impounded in the irrigation ditches.
In such a situation it is impossible to avoid deposition
of the chemical into these waters. As a result, a possible
pollution problem exists since seepage of these waters
through leaky floodgates often occurs, and, in the advent
of heavy precipitation shortly after application, the
water level must be lowered by draining to prevent
prolonged soil saturation or flooding which is injurious
to the cranberry vines. For these reasons, the following
investigation was undertaken.

Materials and Methods
A section of cranberry bog measuring 2900 ft- was
treated with parathion at a rate equal to 1 lb/ acre. The
treated area was completely surrounded by an irrigation
ditch 3 feet wide by 3 feet deep. Water for this ditch

University of Massachusetts, Cranberry Experiment Station, E.
Wareham. Massachusetts 02538.

Present address; U. S Dept. of the Interior, Bureau of Commer-
cial Fisheries, Biological Laboratory, Sabine Island, Gulf Breeze,
Fla. 32561.

was pumped from an adjacent pond up through a drain-
age canal a distance of 200 yards. The bog and the
irrigation ditch surrounding it are separated from the
drainage canal by a roadway 15 feet wide, the waters
passing beneath the roadway in a culvert. A floodgate
on the bog impounded the water in the irrigation ditch
when the water level was approximately 1 to 2 inches
below the bog surface. At this time, the pumping of
water to the bog ceased and the water in the drainage
canal allowed to recede to its normal level. When this
happened, the level of the impounded irrigation water
was approximately l'/2 feet higher than that of the
drainage canal.

The chemical was applied to the test area through
overhead sprinklers when the irrigation ditch was full
and the waters impounded. The sprinkler's pattern was
such that no chemical-containing waters fell in the
drainage canal, but deposition did occur in the irriga-
tion water. Slight runoff of this application water from
the bog surface into the irrigation water was observed.
During the sampling period, seepage of the irrigation
water through the floodgate occurred, lowering the
level approximately 8 inches.

Two 1 -liter water samples were collected, from each of
two locations, from the irrigation waters prior to appli-
cation (controls), immediately following application,
and every 24 hours thereafter for a period of 96 hours.
In addition, similar samples were collected from the
drainage canal at the point where the seeping irrigation
water mixed with the canal waters (bog-canal junction),
and at 50 and 150 yards down from this point using the
same sampling sequence as above.

Vol. 1, No. 2, September 1967

47

The experiment was repeated, with 14 days elapsed
time between the first and second experiment. Data re-
ported herein represent the mean of the two experiments.

Extraction ol the water samples was by the method of
Teasley and Cox (31 Recovery from fortified samples
average Si'"c . and all data have been corrected for
this recovery value. Indentification and quantitation was
made by gas-liquid chromatography using a Varian
Aerograph model 204 equipped with an electron capture
detector. Level of sensitivity was 0.1 ppb. Confirmation
was made by thin layer chromatography. The samples
were spotted on silica gel-coated plates and developed
in chloroform containing 0.7C^ ethyl alcohol. Parathion
was resolved by spraying with palladium ammonium
chloride (0.5 g palladium ammonium chloride and 2
ml cone. HCL in 98 ml distilled water).

Re.sidls and Discussion

Samples from the irrigation ditch collected immediately
after application contained considerable quantities of
parathion (Table 1). The concentration of chemical de-
creased sharply (929f ) after 24 hours, with a subse-
quent reduction of approximately 50% for each suc-
ceeding sample until, after 96 hours, the level was 5 ppb.

TABLE 1. — Parathion concentralions. in ppb. in impounded

irrigation waters and associated drainage waters following

treatment of a cranberry bog'

Location

Time (hours)

0 =

24

48

72

96

Irrigation ditcti

750.0

60.0

25.0

10.0

5.0

(650-850)

(50-75)

(10-35)

(5-20)

(1-9)

Drainaee canal-bog

30.0

3.0

1.0

0.0

0.0

junction

(15.45)

(2.8)

(0.6-1.5)

50 yards down from

0.0 ,

0.3

0.0

0.0

0.0

junction

(0.1-0.5)

150 yards down

0.0

0.1

0.0

0.0

0.0

from junction

(0.08-0.14)

' Values reported are the mean of two separate tests with two samples
per lesl. Figures in parentheses represented the range of the four
analyses.

-■ Immediately following application.

It is possible that the high residue value obtained in the
samples taken immediately after application is a result
of the chemical being somewhat stratified and not uni-
formly distributed throughout the irrigation waters —
the lower concentration detected after 24 hours being a
result of mixing and dilution rather than actual loss.

Santo and Kubo (2) have reported that only trace
amounts of parathion could be lound in irrigation
waters 7 days after application to a rice paddy. The
results reported herein closely correlate with their
findings.

In the associated drainage canal, the presence of para-
thion at a concentration of .10 ppb was also demon-
strated. This indicated that the water seeping through

the floodgate introduced the chemical into the untreated
area, the difTerence in concentration between the two
areas being a result of dilution. After 24 hours the con-
centration in the canal-bog junction had decreased to
3.0 ppb, the rate of disappearance being approximately
the same as that in the irrigation waters. At this time
the chemical was also detected at the locations 50 and
150 yards down from the junction and, although in
lesser amounts, demonstrates movement away from the
point of application. By the end of 48 hours, parathion
could be detected only at the canal-bog junction and by
72 hours the chemical, if present, was below the level
of sensitivity.

The presence of parathion in aquatic environments has
been shown to cause undesirable effects in the associated
biota. Mulla el al. (I) reported a high degree of mor-
tality for mosquito fish [Gamhusia affinis Baird and
Girard) in shallow ponds treated with parathion at rates
of 1 and 0.4 lb/ acre. A 24-hour exposure to a concen-
tration of 65 ppb parathion caused 50% mortality in a
population of sheepshead minnows (Cyprinndnn varie-
i;atiis I.acepede). and a similar mortality value was re-
corded for the brown shrimp (Peiuuiis aztecus Ives)
following a 24-hour exposure to only 5.5 ppb parathion
(4).

It is evident, therefore, that introduction of parathion
into non-target areas can occur under the conditions
described and at concentrations which could be harmful
to certain aquatic organisms. One solution to the
problem would be to remove all waters from the bog
area prior to chemical application. Where this is not
feasible, lowering the water level in the hog to a point
where a sudden heavy rain would not necessitate drain-
ing to prevent vine injury, and insuring absolute water-
tight floodgates will greatly lessen the chances of un-
intentional pollution.

AcknowJedgmenl

This work was supported, in part, b\ funds provided
by the U. S. Department of the Interior as authorized
under the Water Resources Act of 1964, Public Law
88-379.

LITERATURE CITED

(1) Mulla. M. S.. J. O. Keith, and F. ,4. Ounther. 1966.
Pcsistciice and biological effects of parathion residues
in waterfowl habitats. J. Econ. Entomol. ."^Q: 1085-1090.

I2) .Santo. R. and II. Kubo. 1965. The water pollution
caused by organo-phosphoriis insecticides in Japan.
Proc. Second Itit. Water Polliit. Res. Conf. Tokyo. 1964.
Pergamon Press, New York, N. Y. p. 95-99.

(.?) 7(Yj.v/(v, J. I. and W. S. Co.x. 1963. Determination of
pesticides in water by micro-coulometric gas chromatog-
raphy after liquid-liquid extraction. J. Anicr. Water
Works Ass. 5.'i:I09Vl()96.

(4) I'esiicide-Wildliie Stmlies. 1963. U. S. Department of
the Interior Circ. 199.

48

PiiSTiciOKS MoNiroRiNG Journal

GENERAL

Systemic Activity of Zectran, Matacil, and Bidrin Injected
Into Conifer Trunks

John E. Larson', G. R. Pieper', and H. C. Ratsch"

ABSTRACT

Three insecticides — Zeclran® [4-{dimethylamino) 3,5-xyIyl
methyl carbamate], Matacil® {4-{dimcthylainino}-m-tolyl
melliylcarbtiinatc], and Bidrin® (3-hydroxy-N,N-dimethyl-
cis-crolonamide dimethyl phosphate) — were tested for sys-
temic activity using spruce budworm as a bioassay organism.
The materials were injected into the trunks of Douglas fir
and grand fir trees of varying sizes.

In small Douglas fir trees (3 feet or less), movement was
sufficient, and liigli mortality resulted from the injection of
these three compounds at rates of 40 and 200 mg per tree.
In larger trees (5 to 8 feet) treated with 0.2 to 1.0 g of
these chemicals per tree, only Matacil and Bidrin yielded
high mortality. Bidrin gave a higher percentage kill than
Matacil 10 and 17 days after treatment. But after 38 days,
Matacil treatments resulted in liigher percentage kill than
did Bidrin treatments.

Foliage and wood were analyzed for residues of Zectran and
Matacil. Foliage residue levels of 50 ppm and more were
consistently found for Matacil. Foliage residue levels of
Zectran did not exceed 21 ppm in large trees but reached
308 ppm in the small-tree test. Analysis of trunk sections at
points of injection revealed concentrations as high as 8.460
ppm of Zeclran.

Poor results with Zectran were probably the result of its
partitioning into the oleoresin of tree trunks.

THE U. S. Forest Service has underway an extensive
research program aimed at finding safer, more specific
chemical treatments for controlling destructive forest
insects (1). As part of this program, aerial spray tests
with the carbamate Zectran were held on the Bitterroot
National Forest in Montana in 1965 and 1966. The
target insect was the spruce budworm [Choristoneura
fumiferana (Clemens)], an important defoliator.

Along with the aerial test in 1966, a study was also
made of three systemic insecticides injected into tree
trunks. Zectran has previously shown systemic activity
when applied to the soil (2). Besides Zectran. Matacil
and Bidrin were also tested. Matacil is a close analog

Pacific Southwest Forest and Range Experiment Station, Forest

Service, U. S. Department of Agriculture. Bertieley. California

94701.

Commissioned Corps. U. S. Public Health Service, Cincinnati, Ohio

45226.

of Zectran. Bidrin has been reported to have systemic
action in controlling the European elm bark beetle
[Scolytus miiltistriatus (Marsham)], vector of the Dutch
elm disease (3) and of sawfly larvae [Diprion similis
(Hartig)] in eastern white pine (Pinus strobus L.) (4).

Methods and Materials

The test site on the Bitterroot National Forest consisted
primarily of young Douglas fir [Pseudotsuga inenziesii
(Mirh) Franco] and grand fir [Ahies grandis (Dougl.)
Lindl.] at 5,000 to 6,000 feet elevation. Trunk injections
were made in two ways: (a) in trees less than 5 feet, a
small hole was drilled and a tight-fitting glass tube was
inserted; (b) in larger trees, a Mauget injector was used
(5). Silicone rubber, diluted in heptane, was used to form
a leak-proof seal where either device was inserted into
the tree. Normally, Mauget capsules are compressed.
But in our earlier tests in Montana, there were too
many leaks when the capsules were compressed — even
with the silicone rubber seals. Therefore, the Mauget
injectors were used as gravity feeds by drilling a smaller
hole in the lid. In later tests, in which smaller volumes
of insecticide solution were uesd, the Mauget capsules
were compressed successfully.

Nearly all spruce budworm larvae used in the tests
were reared at the Pacific Southwest Forest and Range
Experiment Station, Berkeley, Calif., (Lyon. R. L.. C.
Richmond, and K, Pennington, unpublished data)
where they were fed on artificial media. However,
spruce budworms obtained in the test area were used
during one period in June.

The budworms were caged for 5 days before final ob-
servations were made. Two cages per tree were first
used, but later as many as eight were used. As soon as
evidence of budworm mortality was noted in any of
these cages, additional cages were placed on the tree to
increase sampling accuracy.

Zectran was applied as a 20% or 30% solution in
acetone: Matacil, as a 20% solution in acetone; and
Bidrin, as the technical form (7.9 lbs/gal or 79% w/w).

Vol. 1, No. 2, September 1967

49

The foliage and wood samples for residue analysis of
Zectran and Malacil were eoliected August 10. 1966,
and stored in a eoldroom (35 F) at Berkeley until they
were prepared for analysis. Foliage samples were
analyzed as described by Pieper and Miskus (6). The
wood samples were frozen in liquid nitrogen, then pul-
verized to pass a 30-mesh screen. Fach 3-g sample was
extracted four times with 30-ml portions of benzene for
10 minutes. After each e.Mraciion. the benzene was
decanted and filtered through glass wool with a small
amount of anhydrous Na.jSO,.

To determine the water solubilities of Zectran and
Matacil. each material in excess (about 20 mg Zectran
and 100 mg Malacil) was added to 100 ml water and
adjusted to pH 7.0 by adding three drops of Beckman
3581 concentrated buffer solution. The mixture was
shaken for 21 hours, and excess insecticide was removed
by filtration through a Whatman #1 filter and a #245
Nalgene filter unit (0.45 micron pores). To the Zectran
solution was added 10 ml saturated NaHCO;j. thereby
raising the pH to 9.7. The solution was then extracted
three times with 30-ml portions of benzene.

The Matacil solution was diluted a thousandfold with
acetone and analyzed directly by microcoulometric
analysis for combusted nitrogen.

A partition coefficient for Zectran between oleoresin and
water was determined, using ponderosa pine (Finns
ponderosa Laws.) oleoresin. Five g of oleoresin and 15
ml of double-distilled water were introduced into a
30-ml Squibb-type separatory funnel. To this amount
0.1 mc of Zectran (carbonyl-C") (specific activity of
6.3 mc/mmole) in 5 jxX of methyl cellosolve was added.
After 3 minutes of shaking, the funnel was spun in an
International centrifuge (size 2 240 Head) to partially
separate the tight emulsion formed between the water
and the oleoresin. The water phase was next spun at
97.550 X I.' for 30 min in a Spinco Ultracentrifuge
(Model L. #40 Head). An insignificant number of
oleoresin droplets remained in the water phase. One ml
aliquots of the water phase and 0.1 ml aliquots of the
oleoresin dissolved in toluene were added lo a PPO-
POPOP-naphthalcne-dioxane cocktail and counted in a
Packard Tri-Carb liquid scintillation counter.

Results and Discussion

Before the field test in Montana. Zectran and other
chemicals had been injected into the trunks of small
(less than 12 inches tall) white fir {A. concolor (Gord.
& Glend.) Lindl.] in a greenhouse study at Berkeley.
The Zectran treatments resulted in high percentage kill
for 9 weeks. The degree of kill with Bidrin was less
than that with Zeclran. Applied topically to spruce bud-
worms, Zectran and Matacil were about equally toxic,
and both were nearly twice as toxic as Bidrin. (Lyon.
R. L. persimal tuniiniinicaiion. April 1966). Zectran was

50

next tried in Montana in a large-scale field test on
Douglas fir on May 27, 1966. The trees were divided
into four sizes with three different concentrations of
Zectran and three replications per treatment. The first
spruce budworm bioassay. 2 weeks later, showed poor
kill. In mid-June, a light, natural infestation was found
in all but two of the treated trees. Although some scat-
tered bioassays of these trees were made later, this test
generally resulted in a low degree of mortality. There
was incomplete uptake of the Zectran solution in about
half the treatments. Bioassay data from these trees had
no validity. In the other treatments where uptake was
complete, spruce budworm mortality was still unsatis-
factory. The most likely reason for this failure was the
cool wet weather during June that did not favor rapid
transpiration rates.

Warm dry weather conducive to good transpiration
prevailed from June 27 to July 18. 1966. Several trunk
injections of Bidrin. Matacil. and Zectran were made
during this period on Douglas fir and on grand fir that
ranged from 5 to 8 feet tall. These tests showed clearly
that Bidrin and Matacil gave superior results while re-
sults with Zectran were mediocre (Table 1). No
phytotoxicity resulted from these treatments. In two of
the tests, both the compressed Mauget capsules and the
gravity-type feed were used. There was low mortality in
Zectran-treated trees whether the capsules were com-
pressed or not. Results with Bidrin- and Matacil-treated
trees were sometimes mediocre when the capsules were
compressed — a finding different from what had been
expected.

Residue data from Zectran-treated trees indicated very
poor movement of the chemical from the point of in-
jection, resulting in generally low budworm mortality

TABLE I. — Moilality of spruce budworm on 5- to 8-feet
tall Douglas fir and grand fir resulting from three trunk-
injected insecticides

Appli-
cation

(G/TREE)

Repli-
cations
(No.)

Percent mortality >

(DAYS after treatment)

Chemical

3

10

17

24

38

ZECTRAN

1.0

4

—

33

31

30

14

ZECTRAN

0.6

9

0

25

39

44

—

ZECTRAN

0.2

2

—

—

—

20

—

MATACIl,

1.0

2

—

—

57

89

92

MATACIL

0.2

2

—

—

—

62

—

niDRIN

1.0

5

—

100

82

82

73

BIDRIN

0.2

4

—

—

—

82

—

CONIROL

—

4

0

0

0

0

0

Based upon a 5-d;i.v exposure of an aveiacc of 12 spruce budwornis
(range: 6-24).

Pt^sTKiDEs Monitoring Journal

TABLE 2. — Chenucal residues from 10 injected trees and results of spruce budwonn

bioassy on foliage

Residues (ppm)

Original

AMOUNT

Height

Trunk

OF
TREE

Date

OF

Tree

TION

Final

4 FEET

AT

AT POI 3

No.

Chemical

(G/TREE)

(FEET)

TREAIMENT

BEOASSAY '

Foliage

ABOVE POI -

POI»

(Percent)

1

ZECTRAN

7.15

13.8

5/27

6/8

21.1

4.3

8460

51

2

ZECTRAN

7.15

13.0

5/27

2/6

—

2.7

4170

23

3

ZECTRAN

i.y

5.0

6/27

1/6

7.1

0.0

2820

20

4

ZECTRAN

1.0

5.9

6/27

1/8

1.9

1.1

1900

12

5

MATACIL

I.O

6.0

6/27

6/6

753.0

323.0

456

4.2

6

MATACIL

1.0

6.6

6/27

6/7

50.5

10.5

2320

15

7

ZECTRAN

1.5

10.2

7/6

3/7

1.8

0.0

2770

41

8

ZECTRAN

0.6

8.2

7/18

6/6

T-6

6.0

2470

25

9

ZECTRAN

0.6

7.5

7/18

0/8

r '^n

.10.1

2.0

2670

34

10

ZECTRAN

0.6

7.0

7/18

0/7

r '■'"!

L 10.4 _

67.5

1550

24

Made within 1 week of August 10, 1966, when foliage and wood samples were collected.

T- , No. of dead budworm found after a 5-day exposure

Fraction represents: ■ 1 ^i

Total No. of budworm found after the 5-day exposure

POI = Point of injection. Wood samples collected were 4-mch sections.

Chemical remaining in the 4-inch section.

First figure is residue in top one-third ot crown; second figure is residue in bottom one-third.

(Table 2). From the residues of the eight Zectran-
treatecJ trees, a large amount of Zectran was founci in the
4-inch section at the point of injection. In one instance,
75 days after application, 51% of the original amount
placed there still remained (Tree 1, Table 2). At the
4-inch section 4 feet above this point, only 6 ppm or
less (with the exception of tree #10) was found. There
was a slight accumulation of Zectran found in the com-
parable foliage samples from these trees. This condition
indicates high retention at the point of injection and
implies very little movement in the transpiration stream.
These residues were 99% free, unmetabolized Zectran.

The bioassay data (Table 2) for trees #1 and #8 sug-
gest rather high toxicity of Zectran to budworm. These
bioassays were completed only 3 days before the residue
samples were collected. This anomaly may be attributed
to the too few caging sites used for the insect. In trees
8, 9, and 10 it is evident that the lower one-third of the
crown received more Zectran than did the upper one-
third. These results suggest the importance of the loca-
tion of the cages during the bioassay, and they suggest
the likely pattern occurring in the crown from trunk-
injected materials. Homogeneity was and probably will
always be difficult to attain.

The distribution pattern of Matacil in tree #5 would
seem to answer the question; "What distribution of the

chemical in trees is sought in trunk-injected systemic in-
secticides?" The relatively small amount remaining at
the point of injection (4.2% of the original amount in-
jected) showed much less retention than was true of
Zectran. The superior movement of Matacil in the trans-
piration stream is strongly indicated by the presence of
323 ppm 4 feet up from the point of injection and by
the high accumulation in the foliage (753 ppm). With
this type pattern, much less chemical would give satis-
factory results. That variations occurred even with
Matacil was apparent in the pattern in tree #6, which
showed an intermediate pattern between Matacil-treated
tree #5 and the Zectran-treated trees. But even here, 50
ppm in the foliage gave a high kill.

To explain these differences in the behavior of these
two carbamates, their chemical and physical differ-
ences mav be considered. Matacil had superior mo-
bility in the transpiration stream. In these tests, pene-
tration of the bark, of course, was not necessary
because both chemicals were injected directly into the
sapwood of the trees. The formulations were the same
(20% w/v in acetone), although in certain instances
Zectran was 30% w/v in acetone. When the wood sec-
tions were cut for analysis, a large ring of discolora-
tion appeared in the wood around the point of injection.
This ring of discoloration was also in control trees re-

VoL. 1, No. 2, September 1967

51

ceiving only acetone, and so it can be assumed that the
discoloration was due to the solvent, acetone.

If it is assumed that Zectran and Matacil were carried
with acetone throughout these rings of discoloration,
then both insecticides were exposed to considerable
resin of the ray parenchyma of Douglas fir and grand
fir. In Douglas fir, this region of discoloration would
also include the olcoresin of the resin canals. True firs
of the genus A hies do not normally have resin canals
but may form them in response to wounding. Whether
this happened to the grand fir trees in this study is not
known.

The above consideration would suggest that the more
lipophilic compound tended to partition into the lipid
phase, i.e.. the resin and oieoresin. That this may have
happened for Zectran is further substantiated by its low
water solubility. 100 ppm. and by the fact that its par-
tition coefficient between oieoresin and water was 80:1.
The fact that this partitioning into the lipid phase was
not so likely with Matacil is substantiated by its greater

TABLE 3. — Morlality of spruce biidwonn' after trunk
injections of insecticides into small Douglas fir trees

Appli-
cations

(MG/
TREE)

Repli-
cations
(No.)

Percent mortality

(DAYS after treatment)

Chemical

13

20

27

ZECTRAN

200

2

100

57

100

ZECTRAN

40

1

—

75

100

BIDRIN

200

2

100

100

100

BIDRIN

40

I

—

75

75

MATACIL

200

2

75

100

88

MATACIL

40

1

—

65

100

CONTROL

—

2

0

0

0

Based upon a 5-day exposure to an average of six spruce budworms.

water solubility (1200 ppm). No determination of its
partition coefficient between oieoresin and water has
been made as yet. Since Bidrin is miscible in water, it
is assumed that it would not be tightly bound in the
lipid phase. The low water solubility of Zectran cannot
necessarily be held responsible for its poor ascent in the
larger trees. Herbicides with very low water solubilities,
such as diuron [3-(3,4-dichlorophenyl)-l,l-dimethylurea]
(42 ppm), simazine |2-chloro-4,6-bis(ethylamino)-5-
triazine] (3.5 ppm), and others exert their toxic action
only after ascending in the transpiration stream.

Chemically, the only difference between Zectran and
Matacil is the presence of -CH-, groups in both meta
positions of the benzene ring in Zectran, with only one
meta position being occupied by a -CH, group in the
case of Matacil. The remainders of both compounds are
identical. The single difference chemically does not
affect their toxicity to spruce budworm (i.e., by topical
application) (Lyon, R. L.. personal communication.
April 1966). The differences noted in this study when
both chemicals were trunk-injected, seem to be related
to their different physical characteristics reflected in
their partition coefficients between oieoresin and water.

The foregoing results and discussion of these tests con-
ducted on trees 5 to 8 feet tall gave fairly uniform re-
sults and plausible conclusions. However, a final test
was made in which nine small Douglas fir (less than
3 feet tall) were injected with Zectran. Matacil, and
Bidrin. An attempt was made to duplicate the results
from the earlier, previously mentioned greenhouse test
conducted in Berkeley. Of the nine trees, six each re-
ceived 200 mg of each of the three compounds —
Zectran, Matacil. and Bidrin. One tree treated by each
chemical at this rate was potted and watered regularly
to determine if low soil moisture impeded transpiration.
Three of the nine trees were injected at the rate of 40 mg
of each chemical per tree. All chemicals performed well
at either concentration, and the watering had no effect
(Table 3).

TABLE 4. — Residue analysis of two small Douglas fir injected witli Zectran and Matacil

Dosage

TREE
(MO.)

Heighi

Of

TREE

(INCHES)

Final
uioassay 1

Residue (ppm)

Distance of trunk sections above soil level (inches)

Chemical

0-1

POI =

12-15

Top 10
inches

Foliage »

ZECTRAN
MATACIL

200
200

34.5
26.0

4/4

4/4

23
92

3720
2480

403
113

2
595

131 (308)
437 (631)

Made wilhin 1 week of August 10. 1966. when foliage and wood samples wcie cDlkciid. The fraciicin represents:

No. of dead budworm found after a 5-day exposure

Total No. budworm found after the 5-day exposure. '

P<JI = Point of injection. In Zectran trial. POI was 5- to 6-inch section; n; Matacil. n was 2.5- to .1.5-inch section.

Hirst hKurc is residue determined after November 21. 1966, when wood analysis was made. Figure in parentheses is the caihci dclermiiialioii
made on October I, 1966.

52

Pesticides Monitoking Journal

A residue analysis (Table 4) was made of two entire
trees, one treated with Zectran and the other with Mata-
cil at 200 mg each. The striking diflference here was that
the terminal 10 inches of the Zectran-treated tree con-
tained only 2 ppm, and the same section of the Matacil-
treated tree contained 595 ppm. The foliage analysis
was high in both cases, but the Zectran-treated tree
showed considerable increase in residue from that shown
in the foliage analysis of larger trees. In an attempt to
explain the behavior of Zectran in this test, it may be
pointed out that the distance involved was much shorter
and that young Douglas fir trees contain less oleoresin.
However, the results of this test did confirm the results
of the earlier greenhouse test in Berkeley on trees of
similar size. It should be noted that the results from
small trees in greenhouse tests may be misleading if the
research is later to be applied to a field test.

A root analysis was made only of the Matacil-treated
tree from this small-tree test. The roots were all deeper
than 4 inches below the soil surface. The residue found
was only 1.7 ppm. This low value occurring 35 days
after application indicated essentially no recirculating
by Matacil in the phloem tissue. The ability to translo-
cate in the phloem is considered a very desirable prop-
erty in a systemic chemical. One such compound with
this proprty is the herbicide, amitrole (3-amino-l,2,4-
triazole). {Crisp. C. E.. D. E. Bayer. H. C. Ratsch. and
R. K. Glenn. Comparative tests on the uptake and dis-
tribution of labelled insecticides by Pinus ponderosa,
Pseudotsuga menziesii, and Abies concolor. Unpublished
data on file at Pacific Southwest Forest and Range Ex-

periment Station. Berkeley. California.) Finding a sys-
temic insecticide having both apoplastic and symplastic
mobilities, such as amitrole, could be a significant break-
through in systemic insecticide research.

A cknowledgment

The authors are indebted to Mr. Richard Tierney and
Mr. R. A. Youngberg for their technical help and to
Dr. C. E. Crisp for his critical reading of the manuscript.

The Mauget injectors used were donated by J. J. Mauget
of the Zero Mfg. Co., Burbank, California.

LITERATURE CITED

(!) U.S. Forest Service. 1965. Evaluation of chemicals for
control of forest insects. Pacific Southwest Forest and
Range Exp. Sta., Berkeley, Calif. 8 p., illus.

(2) KenoRa, E. E., A. E. Doty, and J. L. Hardy. 1962.
Laboratory insecticidal tests with 4-dimethylamino-3,5-
xylyl methylcarbamate. J. Econ. Entomol. 55:466-9.

(3) Norris, Dale M.. Jr. I960. Systemic insecticidal action
in the cortical tissue of elm twigs. J. Econ. Entomol.
53:1034-6.

(4) Coppel, Harry C. and Dale M. Norris. Jr. 1961.
Systemii. insecticidal action of certain phosphates in
Finns strobus against Diprion siniilis. J. Econ. Entomol.
54:1061-2.

(5) Coppel, Harry C. and D. M. Norris. Jr. 1966. Bark
penetration and uptake of systemic insecticides from
several treatment formulations in white pines. J. Econ.
Entomol. 59:928-31.

(6) Pieper. G. R.. and R. P. Miskus. 1967. Determination
of Zectran® residues in aerial forest spraying. J. Agr.
Food Chem. (Accepted for publication).

Vol. 1, No. 2, September 1967

53

Problems in Monitoring DDT and Its Metabolites in the

Environment

Donald A. Spencer'

ABSTRACT

DDT is dignidcd lo less harnifiil compounds by a numlicr
oj biolofskal and clicnticcd factors in tlic cnvironiiicnl, which
conversion can take place in as little as a few hours, 'nvolved
are (1) bacteria, soil fungi, and other microorganisms, (2)
enzymatic action, and (3) conversion by reduced porphyrins.
Biological samples should he acquired as quickly after the
death of the organism as possible. If too long a period at
seasonal temperatures has elapsed, there is little value in
reporting the ratio of ntetabo'iles. Biological decomposition
slioiild be arrested by cold s.'orage or dry processing within
a few hoars after collection is made. Avoid anaerobic con-
ditions of storage, even at —20 C. Shorten the period be-
tween collection of sample and chemical analysis. In every
case, report the interval and condition cf storage.

RE-SIDUE problems from persistent pesticides such as
DDT have generated a renewed interest in the means by
which these orpanic compounds can be degraded and
removed from the environment. Research emphasis in
the past 4 years has focused on the role of microorgan-
isms in converting DDT to progressively less toxic
metabolites, and the mechanisms by which man, do-
mestic anim.-iN, fish, and wildlife store, metabolize, and
excrete the DDT-complex. Paradoxically, in the nation-
wide monitoring programs for pesticide residues in the
environment, there is a need to arrest these very same
degradation processes in the interval between the col-
lection of the sample and its eventual chemical analysis.

Bacteria and certain other microorganisms are highly
cfTectivc in converting DDT to TDE, then more slowly
continuing the degradation to simpler compounds. In an
excellent paper recently presented to the Water Pollution
Control Federation Meetings in Kansas City, Hill and
McCarty <l) of -Stanford University studied the degra-
dation of DDT. TDE. lindane, aldrin. dieldrin,
hcptachlor. and endrin by sewage sludge. The active
cultures of anaerobic methane-producing and sulphate-
reducing bacteria produce conditions where fatty acids
are biologically destroyed. DDT was converted to TDE

' Consullanl. National Agricullural Chemicals Association. 11.^5 Fif-
teenth Sirecl. NW.. Washington. D. C. 20005.

almost immediately (DDT was detected only in samples
taken 20 minutes after injection into anaerobic sludge
held at 35 C). TDE then underwent further degradation,
showing a half life of about 4 days. When DDT was
injected into the same culture daily at 1.0 ppm for 57
consecutive days, instead of accumulating, the rate of
conversion of DDT to lower metabolites improved.
Fcllov.ing larger doses of DDT (100 ppm) there was
slower degradation of TDE. ". . . possibh' because a
complexing capacity of the sludge for TDE became
saturated, or because degrading organisms became
poisoned.""

Cope and Sanders (2j became interested in the possi-
bilities of altering the structure of pesticides in water
so as to reduce the hazard to fish. They experimented
with five species of bacteria and found that four of them
(Microitionospora chalrca. Pseudotnonas aeruginosa. P.
fliiorescens. and Corynehacterium pyogenes) "appreci-
ably reduced"" concentrations of DDT in water within
the period of 7 to 16 days. In studies with isotope-labeled
DDT these investigators found that a high portion of
the DDT present was taken up by the bacterial cells or
by contaminating protozoans which were present, and
that one or both microorganisms "apparently metabo-
lized DDT to TDE and possibly to DDE."

At McGill University in Quebec. Barker and Morrison
(3) isolated several microorganisms from the gut of a
DDT-resistant mouse and plated them out on agar-
brain-heart infusion media. When DDT was added and
the cultures incubated for 5 days at 30 C, one isolate,
Proteus vulgaris, dcchlorinated DDT to TDE. Since this
bacterium is one of the primary invaders of animal tis-

Thc clicmical names of compounds mentioned in this paper are:

DDT 1.1.1 •irichloro-2.2-bis( p-chlorophenyl ) ethane

'ri3F: l.l-dichloro-2.2-bis( p-chlorophenyl) ethane

Lindane 1 .2..'l,1..'i.(»-hf\a(hIoror\(lolic\anc, 99% or more gamma

isomer
Aldrin not less than SSf^f of 1.2.3.4. 10.10-hexachloro-I.4.4a,5.8.8a-

hexahydro-1.4-cfK/o-eA'o-5.8-dimethanonaphthalene
Ilieldrin not less than 85% of I.2..1.4.10.10-hex.nchloro-6.7-epoxy-

l.4.4a.S.6.7.8,8a-ociahydro-l,4-CH(/o-exo-5.8-dimethano=

naphthalene
Hcptachlor 1.4.5.6.7.8.8-heptachloro-.1a.4.7,7a-telrahydro-4,7-

mclhanoindcnc
Endrin I.2.'.4.in.lO-hexachloro-6.7-epoxy-1.4,4a,5,6,7,8,8a-

ociahytlro-l,4-('/i(/o-efi(/o-5,8-dimethanonaphthaIene

54

Pesticides Monitoring Journal

sues after death, it is of particular import in the handling
of samples for residue analysis. Mendel and Walton (4).
working in the pharmacological laboratories of the Food
and Drug Administration, Washington, D. C on a
study of the coliform bacteria in the intestines of rats,
clearly demonstrated the role of Escherichia coli and
Aerohacler aerogenes in conversion of DDT. p.p'-DDT
was administered to one series of rats by stomach tube
and to another by intraperitoneal injection. Feces were
collected for 48 hours. From the rats receiving p.p'-DDT
by stomach tube the major chlorinated pesticide in the
feces was TDE with, at most, a trace of DDT. When
these rats were sacrificed 48 hours after dosing, the ratio
of DDT to TDE in the livers ranged from 1:1 to 3:1.
In rats where the gastro-intestinal track was by-passed
by intraperitoneal injection, essentially no chlorinated
pesticides wer& found in the feces collected in the first
48 hours, and the livers of these rats had a 24:1
DDT:TDE ratio. In another phase of the investigation
cultures of E. coli and A . aerogenes were maintained for
24 hours at 37 C resulting in the conversion of 359f to
50% of the introduced DDT to TDE. No more than
30% to 55% of the chlorinated pesticide could be ac-
counted for as DDT or TDE, yet no such loss was re-
corded when- DDT. was incubated with plain culture
medium, indicating that the bacteria had caused the
production of other unidentified degradation products.

Stenersen (5). working at Oslo University for the Nor-
wegian Plant Protection Institute, isolated three bacteria,
Serralia inarcescens, E. coli, and an unidentified strain,
from the excrement of flies. These were grown on bouil-
lon containing C'^-labeled DDT under nitrogen. "All
converted DDT almost completely to TDE (90%) and
DDE (5%)."

In the Department of the Interior's Fish Pesticide
Laboratory at Denver, Wedemeyer (6) chose the facul-
tative anaerobe, A. aerogenes, for detailed studies.
Grown under anaerobic conditions in trypticase-soy
broth containing 5.0 ppm DDT, this bacterium effects
up to 80% conversion to TDE. In a refinement of the
study, A. aerogenes bacteria were disrupted sonically,
resulting in a cell-free system. Under these conditions
the average conversion to TDE was about 70%. leading
Wedemeyer to conclude that reduced cytochrome oxid-
ase is probably the cellular agent in reductive dechlori-
nation of DDT to TDE.

Johnson (16) at the Bureau of Commercial Fisheries,
Biological Laboratory, has now extended our informa-
tion on bacterial degradation by studies on the marine
bacteria (Pseudomonas piscicida). The uptake of DDT
from culture media by this bacteria is very rapid. Within
48 hours, TDE and DDE within the cell walls of the
bacteria constitute 25% of the total DDT-complex. Also
significant in this study of isotope-labeled DDT is the
finding that both bacteria and oysters are capable of

metabolizing small fractions of DDT so completely that
it becomes part .of the metabolic pool and is utilized in
the biochemistry of the cells.

Chacko, Lockwood, and Zabik (7) at Michigan State
University in 1966. demonstrated for the first time the
ability of certain aerobic soil fungi ( Actinomycetes:
Nocardia erythropolis, and five species of Streptomyces)
to convert DDT to TDE. The test organism was cul-
tured for 6 days in a nutrient medium containing 5 to
10 |U,g/ml DDT resulting in 25% conversion of DDT
to TDE in 6 days.

Johnson ct al. (17) found that 23 out of 27 pathogenic
and saprophytic bacteria associated with plants could
convert from a trace to better than 50% of p,p'-DDT
to p,p'-JDE in a space of 2 weeks under anaerobic con-
ditions. In the majority of cases, the pace of conversion
quickened during the second week.

Clear Lake in California is a large, relatively shallow,
warm body of water (41,600 surface acres) with bottom
deposits of soft, deep, black ooze. The lake is rather
turbid most of the time (8). Miskus, Blair, and Casida
(9) in 1965 collected a sample of this lake water near
Lakeport, introduced ring-labeled C'^-DDT at 0.01 ppm
and incubated it in a stoppered flask for 7 days at room
temperature. In the I -week time, 70% to 80% of the
DDT was converted to TDE. The results were verified
by two additional methods. Six additional samples were
collected later from various parts of the lake and
similarly tested. These samples varied markedly in the
amounts of DDT converted to TDE, but the two samples
that contained large amounts of plankton converted
83% and 95%, respectively, of DDT to TDE within
the week.

To this point, the importance of biological systems in
the degradation of DDT has been stressed. There is,
however, no agreement that this is the only factor, or
even the most important. Ecobichon and Saschenbrecker
(10). working at the University of Guelph in Ontario,
Canada, repeated the analysis of a frozen sample of
avian blood that they had studied 3 weeks before, by
mistake. The ratio of DDT to its metabolites TDE and
DDE were so different from their original readings that
it presented a possible source of error in analytical
procedures. They then prepared a single large sample
of heparinized chicken blood, introduced an acetone
solution of technical DDT at 1.0 ppm, sealed the flask
and stored it at —20 C. Each week the sample was
removed, quickly thawed, a 2-ml subsample removed
for analysis, and the basic sample returned to —20 C.
This was repeated for 12 consecutive weeks. Both p,p'-
DDT and o.p'-DDT completely disappeared by the 10th
week while DDE and TDE increased in quantity until
the 7th week and then in turn began to slowly decrease.
At the same time, a plasma sample that contained DDT
serving as a control showed no evidence of degrada-

Voi.. 1, No. 2, September 1967

55

lion. Wiih the repeated freezing and thawing the
eryihrocyles in the hlood were hcniolyzcd. thus expos-
ing the insecticides to high concentrations of free
hemoglobin. The authors suggest that tissues and
microorganisms which contain large quantities of re-
duced coenzymes, porphyrins, and other metaliopro-
teins could carry out these steps by simple chemical
redox reaction.

Castro (11) at the University of California, Riverside,
exposed dilute solutions of iron porphyrins (Fe+ +
deuterioporphyrin) to DDT at room temperatures. The
porphyrin complex was rapidly oxidized. Castro points
out that iow-valent iron porphyrin complexes are mani-
fest in all aerobic organisms.

Miskus, el at. (9) at the University of California,
Berkeley, were also interested in the role of reduced
porphyrins. They added C"-labeled DDT to solutions
of hemoglobin or of hematin in a Thunberg tube and
shook the mixture for 4 hours at room temperature. No
conversion took place unless the color remained red,
representing the state of reduced porphyrins. By adding
sodium dilhionite to the hemoglobin mixture the con-
version of DDT to TDE ranged from 60% to 75%.

At the Monks Wood Experimental Station in England,
JefTeries and Walker (12) were studying the acute and
chronic toxicity of p.p'-DDT to Bengalese finches by
feeding caged birds concentrations of pure DDT in their
diet. Birds that died, or were sacrificed at different pe-
riods, were placed in ~1 1 .5 C to — 14.5 C refrigeration.
The livers of two treated birds were analyzed within 10
minutes of death and the ratio of DDT:TDE was 100:1,
a negligible conversion. Thereafter at different intervals,
groups of treated birds were withdrawn from refrigera-
tion, dissected, and the livers analyzed. By the 67th day
in cold storage, the conversion of DDT to TDE was
1:1 — fairly convincing evidence that cold storage of a
little over 2 months permits significant changes in the
ratio of DDT with that of its metabolites.

At Mississippi State University. Walley, Ferguson, and
Culley (13) have attempted to segregate the chemical
and bacterial factors as they pertain to the degradation
of DDT in the liver in vitro. Livers were removed by
sterile techniques from newly sacrificed birds, the livers
sliced and transferred to cultural vials containing a
sterile medium in which 50 fxg of purified p.p-DDT had
been added. Incubated at 37 C. subsamples showed the
presence of TDE and traces of DDE by the end of 24
hours. By 96 hours much of the DDT had been con-
verted to TDE. No bacterial contamination could be
demonstrated in the liver cultures at the end of the study.
Again, the capability of tissues, independent of living
microorganisms, to convert DDT to lower metabolites is
indicated. Basically there are three factors responsible
for the degradation of p.p'-DDJ. and the shift in ratio
between metabolites: (1) the continuing activity of

bacteria and other microorganisms, (2) enzymatic ac-
tion, and (3) conversion by reduced porphyrins. The •
activity of bacteria and reduced porphyrins to convert
DDT is greatly enhanced by anaerobic conditions —
which is quite characteristic of the handling of many
samples for residue analysis.

Most programs today that monitor pesticide residues
in the environment attempt not only to analyze for
DDT, TDE, and DDE, but both the para-para and
ortho-para isomers of all three. The cost of chemical
analysis and the time required to make such studies
is appreciably greater than simply searching for p.p'
-DDT. Nevertheless there is considerable merit in the
new approach. Not only do the metabolites of DDT have
less toxicity for man, domestic animals, and wildlife,
but the ortho-para isomer of DDT is also less toxic by a
factor of 5 to 9 times in tests with rats (14). These six
pesticides also differ in their persistence in the environ-
ment, the rate of storage in neutral fat, the elimination
of residue from fat depots, and the routes by which they
are degraded (J5).

The problems of monitoring the DDT-complex in the
environment begin with the choice of samples. Too
often samples are collected an unknown time after the
death of the organism, when it was the actual meta-
bolites present at the time of death that were important.
For example, a fish kill is reported in a stream draining
cotton fields on which pesticides have been used. Fre-
quently it is several days to a week before responsible
investigators reach the scene to collect samples. At
summer temperatures, conversions of the insecticide
progress very quickly as some of the foregoing labora-
tory studies indicate. In other cases the program may
make use of parts of game animals contributed by
hunters from broad sections of the country. There is
considerable lag at seasonal temperatures before these
samples can be properly stored or processed. Bttt it
can also be the deliberate action of an investigator who
eventually collects specimens of eggs "one week past
expected hatching date in advanced state of decomposi-
tion." There is little point to reporting metabolites in
fractional parts per million at this late date if the study
is concerned with the possible effect on hatching success
of the egg.

However, most samples collected for monitoring pesti-
cide residues correctly reflect the ratio of metabolites
at the time: silts from a stream bed, muck from a tidal
marsh, water and its suspended organic matter, vegeta-
tion, pools of small invertebrates, even blood and fat
samples taken by biopsy. But the collector is commonly
4 to 8 hours away from facilities for processing or
otherwise storing his samples. Deep freeze is com-
monly employed for checking any further spoilage of
biological samples; however, certain bulky samples such
as soil, muck soils, water with suspended organic matter,

56

Pesticides Monitoring Journai,

and vegetation often lack this protection. In fact, some
bottom sediments, sealed in metal or glass containers
have exploded from gases generated by anaerobic bio-
logical action before the sample could be handled by
the chemist. Lastly, the field collection commonly is a
seasonal matter, and numbers of samples sufficient to
keep a residue analysis laboratory busy for a whole year
are collected in a matter of weeks. It is a fact that
samples have been held 1 to 2 years in crowded chest-
type refrigerators awaiting analysis.

(16) Johnson. Robert F. 1967. Food chain studies. USDI,
Bur. Commercial Fish. Circ. 260, p. 9-11.

(17) JohiLsoii, B. Thoma.^, Robert N. Goodman, and Herbert
S. Goldberg. 1967. Conversion of DDT to ODD by
pathogenic and saprophytic bacteria associated with
plants. Science 157(3788):560-561.

LITERATURE CITED

(J) Hill. David W., and Perry L. McCarly. 1966. The
anaerobic degradation of selected chlorinated hydro-
carbon pesticides. Presented: Water Pollut. Contr.
Fed. Meeting, Kansas City.

(2) Cope, O. B., and H. O. Sanders. 1963. Microorganisms
and hydrocarbons. USDI: Fish and Wildlife Circ,
Pesticide-Wildlife Studies. 167:27.

(3) Barker, P. S., and F. O. Morrison. 1965. The metab-
olism of TDE by Proteus vulgaris. Canad. J. Zool.
43:652-654.

(4) Mendel, J. L., and M. S. Walton. 1966. Conversion
of pp'-DDT to pp'-DDD by intestinal flora of a rat.
Science 151(3717): 1527-1528.

(5) Stcnersen, J. H. V. 1965. DDT-metabolism in resistant
and susceptible stable flies and in bacteria. Nature
(London) 207(4997):660-661.

(6) Wedemcyer, Gary. 1966. Dechlorination of DDT by
Aerobacter Aerogenes. Science 152(3722):647.

(7) Chacko, C. /., J. L. Lockwood, and M. Zabik. 1966.
Chlorinated hydrocarbon pesticides: Degradation by
microbes. Science I54(3750):893-895.

(8) Hunt, Eldridge G., and Arthur I. Bischoff. 1960.
Inimical effects on wildlife of periodic DDE applica-
tions to Clear Lake, Calif. Fish and Game 46(1):
91-106.

(9) Miskiis. Raymond P., Deanna P. Blair, and John E.
Casida. 1965. Conversion of DDT to DDD by bovine
rumen fluid, lake water, and reduced porphyrins.
J. Agr. Food Chem., 13(5):48I-483.

(10) Ecobichon, D. J., and P. W. Saschenbrecker. 1967.
Dechlorination of DDT in frozen blood. Science
156(3775):663-665.

(11) Castro, C. E. 1964. The rapid oxidation iron (Fe")
porphyrins by alkyl halides. A possible mode of intoxi-
cation of organisms by alkyl halides. J. Amer. Chem.
Soc. 86(11):2310-2311.

(12) Jefferies. D. L., and C. H. Walker. 1966. Uptake of
Pp'-DDT and its post mortem breakdown in the avian
liver. Nature (London) 212(5061):533-534.

{13) Walley, W. Wayne. Denzel E. Ferguson, and Dudley
D. Culley. 1966. The toxicity, metabolism, and fate
of DDT in certain icterid birds. J. Miss. Acad. Sci.
XII:281-300.

(14) Dale. W. £., M. F. Copeland. G. W. Pearce, and J. W.
Miles. 1966. Concentration of op'-DDT in rat brain
at various intervals after dosing. Arch. Int. Pharma-
codyn. 162(l):40-43.

(15) Menzie, Calvin M. 1966. Metabolism of pesticides.
USDI Fish and Wildlife Serv., Spec. Sci. Rep. No. 96,
274 p.. May.

Vol. 1, No. 2, September 1967

57

Infornuilion for Contributors

The Pesticides Monitoring Journai 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
sampling 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-
laboratory checks. The procedure employed should be
referenced or outlined in brief form, and crucial points
or modifications should he 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 Styi E Manual for Biological
JoLRNAi s. American Institute of Biological
Sciences, Washington, D. C, and/or the Styi e
Manual of the United States Government Print-
ing Office.

. An abstract (not to exceed 200 words) should

accompany each manuscript submitted.

All material should he submitted in duplicate

(original and one carbon) and sent by first-class
mail \n flat form — not folded or rolled.

Manuscripts should he typed on 8' i x I 1 inch
paper with generous margins on all sides, and
each page should end with a completed para-
graph.

All copy, including tables and references, should

be double spaced, and all pages should be num-

bered. The first page of the manuscript must
contain 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 alpha-
betically, giving name of author/s/, year, full title
of article, exact name of periodical, volume, and
inclusive pages.

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 lhe\' 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 publishetl
or submitted for publication elsewhere, notation of such
should he pro\ided.

Correspondence on editorial and circulation matters
should he addressed to: Mrs. Sylvia P. O'Rear. Editorial
Manager, Pesticides Monitoring Journal, Pesticides
Program, National Communicable Disease Center, At-
lanta, (ieorgia .10.VVV

58

Pesticides Moniiorinc Journai.

The Pesticides Monitoring Journal is published quarterly under the auspices of the Federal
Committee on Pest Control and its Subcommittee on Pesticide Monitoring as a source of
information on pesticide levels relative to man and his environment.

The parent committee is composed of representatives of the U. S. Departments of Agriculture,
Defense, the Interior, and Health, Education, and Welfare.

The Pesticide Mentoring Subcommittee consists of representatives of the Agricultural Research
Service, Consumer and Marketing Service, Federal Extension Service, Forest Service, Depart-
ment of Defense, Fish and Wildlife Service, Geological Survey, Federal Water Pollution
Control Administration, Food and Drug Administration, Public Health Service, and the
Tennessee Valley Authority.

Responsibility for publishing the Pesticides Monitoring Journal has been accepted by the
Pesticides Program of the Public Health Service.

Pesticide monitoring activities of the Federal Government, particularly in those agencies repre-
sented on the Pesticide Monitoring Subcommittee which participate in operation of the national
pesticides monitoring network, are expected to be 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 nongovernment research
institutions, both within and without the United States. 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 Subcommittee. Authors are given the benefit of review comments prior to
publication.

Editorial Advisory Board members are:

Reo E. Duggan, Food and Drug Administration, Cliairman

Andrew W. Breidenbach, Public Health Service

Anne R. Yobs. Public Health Service

James B. DeWitt, Fish and Wildlife Service

S. Kenneth Love, Geological Survey

Milton S. Schechter, Agricultural Research Service

Paul F. Sand, Agricultural Research Service

Trade names appearing in the Pesticides Monitoring Journal are for identification only and do
not represent endorsement by any Federal agency.

Address correspondence to:
Mrs. Sylvia P. O'Rear
Editorial Manager

PESTICIDES MONITORING JOURNAL

Pesticides Program

National Communicable Disease Center

Atlanta, Georgia 30333

CONTENTS

Volume 1 December 1967 Number 3

Page

EDITORIAL

Units for reporting pesticides 1

S. K. Love

RESIDUES IN FOOD AND FEED

Chlorinated pesticide residues in
fluid milk and other dairy products

in the United Slates 2

R. E. Duggan

RESIDUES IN FISH, WILDLIFE,
AND ESTUARIES

Chlorinated pesticide levels in the

eastern oyster (Crassostrea virginica)

from selected areas of the South

Atlantic and Gulf of Mexico 9

John C. Bugg, Jr., James E. Higgins,
and Eric A. Robertson, Jr.

Galveston Bay pesticide study — water
and oyster samples analyzed for
pesticide residues following mosquito

control program 13

Victor L. Casper

Investigation of effects of large-scale
applications of 2,4-D on aquatic fauna

and water quality 16

Gordon E. Smith and Billy G. Isom

PESTICIDES IN SOIL

Monitoring for chlorinated hydrocarbon
pesticides in soil and root crops in the

Eastern States in 1965 22

W. L. Seal, L. H. Dawsey, and G. E. Cavin

EDITORIAL

Units for Reporting Pesticides

The choice of units for reporting pesticide residues and
concentrations is largely arbitrary but is influenced by
custom, the magnitude of values commonly measured,
and the nature of the environment.

Custom is perhaps the strongest influence in scientific as
well as in lay circles. If residues in soils are customarily
reported in parts per million, the tendency will be to
continue the practice unless convincing reasons are pre-
sented to change to other units.

The magnitude of values commonly measured generally
dictates the size of the unit selected but not the system.
For example, milligrams per kilogram (mg/kg) will
serve very well for values in the range of 1 to 1,000 mg.
but micrograms per kilogram (^xg/kg) would be better
if the values are in the range of 0.001 to 0.1 mg which
is equivalent to 1 to 100 /xg. However, choice of the
metric (dimensional) system of milligrams and micro-
grams per kilogram rather than the nonmetric (dimen-
sionless) system of parts per million and parts per
billion is an arbitrary decision based on custom or
preference.

It has been pointed out that parts per million can be
considered metric or nonmetric. This is true. For sim-
plicity in this discussion, however, milligratns, micro-
grams, etc., will be considered metric and parts per mil-
lion, parts per billion, etc., nonmetric.

The nature of the environment for which pesticides are
being reported is also a factor in the choice of units. For
example, in the water environment, concentrations gen-
erally are measured in units from one to two orders of
magnitude smaller than those measured in soils, food,
and fish and wildlife. Thus, the principal Federal agencies
that measure pesticides in water have chosen micrograms
per liter (/j,g/l) as the primary unit for reporting pesti-
cide concentrations. Micrograms per kilogram (/j,g/kg)
is used for concentrations in water-associated sediments.
A canvass was made of the Federal agencies represented
on the Monitoring and Research Subcommittees of the
Federal Committee on Pest Control to ascertain presently
used units for reporting pesticide residues and concen-
trations. Of 12 respondents, 6 preferred using metric
units, 3 preferred nonmetric units, and 3 had no strong
preference.

Those agencies that prefer nonmetric units have frequent
contacts and dealings with nontechnical people who have
become accustomed to these units. There is strong reluc-
tance to convert to different units that might cause con-
fusion. This attitude may be valid for the short haul.
However, if the United States is ever going to change
to metric units for all measurements, it seems clear that
both public and private agencies and groups will have to
provide examples by getting on the bandwagon.

The Editorial Board of the Journal is not contemplating
requiring authors to report pesticide residues and con-
centrations in any particular system or unit. Authors
know, or should know, what units are best suited to the
profession and to the reader audience. However, the
Board strongly recommends the use of metric units
wherever possible. Furthermore, to aid in transition from
nonmetric to metric, it will be acceptable practice to
report values in metric followed by nonmetric values in
parentheses. For example, 200 mg/1 (200 ppm). The
decision as to this form of expression versus the cus-
tomary practice will be the author's.

More than one system of units or orders of magnitude in
a single system should be avoided in a given paper. It is
particularly confusing to use two or more orders of mag-
nitude in a single table. For example, micrograms per
liter (/ig/l) and milligrams per liter (mg/1) should not
be used in the same table. Erroneous impressions are
easily formed by the reader in such instances.

Metric (dimensional) units have the advantage of show-
ing actual weights of pesticides, whereas nonmetric (di-
mensionless) units do not. Knowledge of oiiual weights
of specific pesticides in an environment is significant to
many investigators. Nearly all continental European
countries and many others throughout the world use
metric units.

Views of Journal readers will be welcomed by the Editor.
If there is sufficient reader interest in this and other topics
related to monitoring pesticides, it may be desirable to
include a section in the Journal on "Communications to
the Editor."

S. K. Love

Member, Editorial Advisory Board

Vol. 1, No. 3, December, 1967

RESIDUES IN FOOD AND FEED

Chlorinated Pesticide Residues in Fluid Milk and Other Dairy Products in the United States

R. E. Duggan'

ABSTRACT

The findings on 12,836 objective samples of milk and dairy
products examined by the U. S. Food and Drug Administra-
tion from domestic and imported lots during the period July
I, 1963, through June 30, 1966, are reported. A majority
of the samples contained pesticide residues. Residues of
DDT, DDE, TDE, dicldrin. heptachlor epoxide. BHC, lin-
dane, aldrin, heptachlor, and methoxychlor account for
99.3% of the residues. About 95% of the values were below
0.51 ppm on a fat basis, and 71.5% of the values were
below 0.11 ppm on a fat basis. The average level for DDT
and its analogs was 0.134 ppm on a fat basis, slightly more
than one-tenth the legal tolerance of 1.25 ppm for the
combined DDT compounds. The average levels for dietdrin
and heptachlor epoxide were 0.042 and 0.036 ppm, fat basis,
respectively — slightly more than one-tenth of the current
administrative guides for each chemical. The levels and
kinds of pesticides are in good agreement with the findings
on total diet samples examined during this period.

Introduction

Recently, tolerances were established for DDT and its
analogs, singly or combined, at levels of 0.05 ppm in
fluid milk and at 1.25 ppm in the fat of other dairy
products. Tolerances were requested by a petition sub-
mitted by the State of California. Following a review of
the petition and evaluation of other data, a Committee
appointed by the National Academy of Sciences re-
commended that these tolerances be established.

In 1957 and 1959, Clifford et at.. (I. 2) reported results
of surveys by the Food and Drug Administration on resi-
dues of pesticides in market milk. Since that time, major
advances in gas-liquid chromatography and other im-
provements have been made in the methods of analysis
used to determine the kind and quantity of residues in
milk. Thus, we do not consider the earlier data com-
parable to current findings because of significant changes
in analytical procedures.

' Deputy Associalc Commissioner for Compliance, Food and DruR
Adminlsiralion, U. S. IJcpanmcnt of Health, Education, and Wel-
fare. Washington, D. C. 20204

The principal purpose of this paper is to report and eval-
uate the findings on 8,548 samples of fluid milk and
3,598 samples of other dairy products e.xamined by the
U. S. Food and Drug Administration between July 1,
1963, and June 30, 1966, within the United States. The
findings on 690 samples of manufactured dairy products
imported into the United States from 29 countries and
examined for pesticide residues are also reported.

Sampling Procedures

Samples of fluid milk and manufactured dairy products
were collected nationwide as a part of FDA's surveil-
lance program on pesticide residues in foods. The pro-
gram predetermined the total number of samples to be
collected at each of its 18 District Laboratories but did
not specify how many of these were to be "objective"
versus "selective." The selection of sampling points and
the scheduling of samples also was left to the discretion
of the District offices. In the surveillance program, sam-
ples classified as "objective" are unknown with respect
to suspicion of residue content or actual misuse of pesti-
cide chemicals. Samples collected because of suspected
excessive residues are classified as "selective." Only the
"objective" samples are included in this report.

Fluid milk samples were collected from bulk tank trucks,
bulk storage tanks at milk and dairy product plants, and
from stocks of bottled milk. Therefore, each sample
represents composites from one or more dairy herds.
The samples of domestic manufactured dairy products
(butter, cheese, condensed milk, frozen desserts, and
other products) were collected under similar program
instructions.

Samples of imported dairy products were collected from
shipments at the time the products were offered for entry.

The physical sample was taken after mixing fluid milk,
or by removing several portions of solid products, for
compositing in the laboratory. Several units of products
in containers were collected as a sample. Where codes
or balch numbers were used, each batch was sampled
separately.

Pesticides Monitoring Journal

Samples were collected from 45 States in FY 1964, 47
States in FY 1965, and from 42 States in FY 1966.
Samples were collected from the District of Columbia
each year. No samples were obtained from Alaska, and
only one sample was reported from Hawaii.

Analysis

Generally, samples were examined promptly after col-
lection.

All analyses were performed in FDA District Laboratoines
using multi-residue gas-liquid chromatographic methods.
Microcoulometric and electron capture detectors were
employed. The official A.O.A.C. method (3) to detect
multi-residues was used in FY 1965 and 1966. The quan-
titative sensitivity of 0.25 ppm (fat basis) was based on
V2 full-scale deflection (1 x 10'^ AFS) for 1 nanogram
of aldrin. These procedures are described in detail in
FDA's Pesticide Analytical Manual — Volume I (4). Resi-
dues above these sensitivity levels were confirmed by thin
layer chromatography. Quantitative figures are reported
below these sensitivities but were not confirmed by check
analysis and are recognized as having reduced accuracy
common to all quantitative estimations at the lower
ranges of method sensitivity. All FDA laboratories par-
ticipated in a collaborative study (5) of the method

using heptachlor epoxide and dieldrin. This study showed
an average recovery of 113% for heptachlor epoxide and
95.9% for dieldrin. Standard deviations of ±0.039 ppm
at the 0.29 ppm level for heptachlor epoxide and ±0.052
ppm at the 0.26 ppm level for dieldrin were reported.
Additional data are being published (6) describing the
application of this method to other pesticide chemicals.
All results are reported on a fat basis, and no correction
for recovery has been made.

Results

Although a majority of the samples were collected within
milk-producing States, some were not. In order to eval-
uate the distribution of the sampling, the samples were
grouped in Table 1 according to the U.S.D.A. Crop Re-
porting Divisions for comparison with milk production
(7).

A total of 12,836 samples were collected and examined
during the 3-year period, distributed by year and product
class as shown in Table 2.

Residues were reported in 7,346 (57%) of the total
samples examined. More than one pesticide chemical was
found in 5,154 samples.

The percent of samples containing residues and multiple
residues are shown for each year in Table 3.

TABLE 1. — Comparison of samples and incidence of residues with production of milk

Total Samples

Percent Distribution of

Samples

Percent
Production

Percent

Percent
containinq

By Fiscal Year

FY 1966

Distribution

RESIDimS

1964

1965

1966

North Atlantic

19.3

16.1

74.3

12.8

17,2

20.8

E. North Central

28.8

27.9

30.3

29.3

23.9

29.6

W. North Central

20.9

15.6

48.7

16.4

13.2

17.0

S. Atlantic

7.0

13.4

68.0

12.8

15.9

11.5

S. Central

10.6

15.4

74.2

14.9

17.1

14.2

West

13.3

11.7

77.6

13.8

12.7

6.8

1 U. S. Department of Agriculture, Crop Reporting Divisions.

TABLE 2. — Distribution bv year and product class of

12,836 samples collected and examined during fiscal years

1964-1966

TABLE 3. — Percent of samples, by fiscal year, containing
residues and multiple residues

Percent of Total Samples

Samples

1964

1965

1966

DOMESTIC:

Fluid Milk

28.7

18.6

19.3

Mfd. Dairy Products

14.9

8.4

4.8

IMPORTED:

Mfd. Dairy Products

2.1

1.3

1.9

TOTAL

45.7

28.3

26.0

Samples

Percent Samples
with RESiDires

Percent Samples

WITH

Multiple Residues

1964

1965

1966

1964

1965

1966

DOMESTIC:

Fluid Milk

41.0

72.8

69.3

24.9

51.6

50.7

Mfd. Dairy Products

47.8

68.1

62.8

36.2

52.4

42.9

IMPORTED:

Mfd, Dairy Products

38.0

55.8

60.4

22.4

33.7

45.6

Vol. 1, No. 3, December, 1967

Ten chemicals— DDE, dicldrin, DDT, heptachlor epox-
ide, TOE, BHC, lindane, aldrin, heptachlor, and me-
Ihoxychlor— account for 99..irr of Ihc residues. Twenty-
three other pesticide chemicals representing 131 residues
also were found. However, except for pentachlorophenol
found in 20 samples, these were found too infrequently
to be considered significant.

Table 4 shows the incidence of the above 10 specific
residues in percent of total samples. Since more than
one residue is found in many samples, the total exceeds
100%. The factor for number of residues per sample
was 1.5 based on all samples and 2.6 based on the posi-
tive samples only.

Table 5 shows the percent of residues at arbitrarily se-
lected ranges in levels of residues and is based on the
total number of residues of the specific chemical found.
The percent of residues in the various ranges was rela-
tively uniform between fluid milk and manufactured
dairy products, both domestic and import. There was a
definite break between the range 0.11-0.50 ppm and the
next higher range of 0.51-1.00 ppm. Ninety-five percent
of all residues were below 0.51 ppm, and 71.5% were
below 0.1 1 ppm.

The average pesticide level for each chemical shown in
Table 5 includes all samples and was calculated by using
the mid-point of each range and the percent of samples
falling in the range. 1 he actual average values were used
for the range exceeding 2.00 ppm. The standard devia-
tion and 95'^'r confidence limits are shown for each
chemical. The large standard deviation is not unexpected
because of the large number of negative samples. The
large number of negative findings and low values must
be considered in using the standard deviation.

Table 6 shows the percent distribution of residues, by
year and product class, in difTercnt quantitative ranges.

This information is shown for total residues, as well as
for residues of individual chemicals.

Generally, there arc no significant changes in the inci-
dence and relative levels of residues when individual
chemicals are considered on an annual or commodity
basis.

TABLE 4. — Frequency of specific chemicals,
July I, 1963 — June 30, 1966

Percent

OF Samples Containino Residues

Pesticide

Total

Milk

Other Dairy Products

Domestic

Import

DDE

40.6

40.7

41.8

34.3

Dieldrin

27.9

30.0

26.2

17.0

DDT

24.t

23.5

25.3

24.5

Heptachlor epoxide

23.6

25.4

23.2

3.2

TDE

16.9

15.5

20.6

16.4

BHC

8.0

6.7

9.6

15.5

Lindane

5.6

4.9

7.1

5.9

Aldrin

1.4

1.6

0.5

3.9

Heptachlor

0.9

1.0

0.8

0

Methoxychlor

0.7

1.0

0.6

0

DDE

Dieldrin

DDT

Heptachlor epoxide

TDE
BHC

Lindane

Aldrin

Heptachlor

Metho.xvthlor

l,l-dichloro-2,2-bisCp-chIorophenyI)ethyIene

not less than 85'!^ of l,2,3,4.10.10-hexachloro-6,7-

epoxy-1.4.4a.5,6,7.8.8a-octahydro-l,4-enJo-exo-5,8-

dimeihanonaphthalene

l.l.l-trichloro-2.2-bi5;{p-chlorophenyn ethane

1.4. 5.6.7,8, 8-heptachIoiu-2,3-epoxy-3a.4,7.7a-

tetrahydro-4,7-methanoindan

l,l-dichIoro-2,2-bis{p-chlorophenyl) ethane

1,2,3,4,5.6-hexachlorocyclohexane, mixed isomers

1.2,3,4,5.6-hexachlorocycIohexane, 99^c or more

gamma isomer

not less than 95*?^ of 1,2,3,4,10,10-hexachloro-l,

4.4a.5.8,8a-hexahydro-1.4-i';u/n-p,vo-5.8-dimethano=

naphthalene

1.4.5. 6.7.8. 8-heptachloro-3a.4.7.7a-tetrahydro-4,7-
methanoindene

l.l,l-trichloro-2.2-bis{p-methoxyphenyl) ethane

TABLE 5. — Percent distribution of residues of

specific chemicals in different ranges — 3-yea

- total

Percent Distribution of Positive Samples

(ppm)
(fat basis)

All
Residues

DDT

DDE

TDE

Dieldrin

Hepta-
chlor
epoxide

BHC

Lindane

Aldrin

Hepta-
chlor

Methoxy-
chlor

T-0.03

45.5

48.2

43.8

51.6

38.2

36.3

62.1

67.1

84.3

66.1

24.7

0.04^.10

26.0

23.6

26.6

28.0

28.7

27.8

19.7

20.7

8.4

18.3

17.2

0.11-0.50

25.3

24.0

25.9

17.7

30.0

33.1

16.1

10.4

7.3

14.8

44.1

0.51-1.00

2.3

i.6

2.6

1.4

2.6

2.2

1.8

1.4

—

0.9

7.5

1.01-1.50

0.5

0.6

0.7

0.5

0.2

0.5

0.3

0.3

—

—

3.2

1.51-2.00

0.2

0.3

0.2

0.1

0.1

0.1

—

—

—

—

2.2

>2 <^

0.3

0,7

0.2

0.7 n.i

<0.03

—

0.1

—

—

1.1

AvcraKc'

0.042

0.066

0.026

0.042

0.036

0.007

0.004

0.001

0.0O2

0.001

Standard

Deviation'

0.278

0.358

0.264

0.138

0.110

0.049

0.042

0.010

0.045

0.013

95% Con

Odcncc Limiii

±0.005

±0.006

±0.005

±0,002

±0.002

±0.0009

±0.0007

±0.0002

±0.0008

±0.0002

'All Hinpic] were used to calculate the averages, standard deviations, and confidence limits.

4

Pesticides Monitoring Journal

TABLE 6. — Percent distribution of residues, by fiscal year and product class, in different quantitative ranges

Percent Distribution of Positive Samples

Ranoe

Fluid Milk

Manufactured Dairy Products

(PPM)
(FAT BASIS)

Domestic

Domestic

Imported

1964

1965

1966

1964

1965

1966

1964

1965

1966

I. ALL RESIDUES

T-0.03

41.5

44.5

49.7

43.5

48.3

47.8

44.0

35.8

42.9

0.04-0.10

26.6

25.7

24.8

28.7

24.4

26.4

22.9

31.1

25.4

0.11-0.50

27.6

26.2

22.0

26.0

25.3

24.5

27.7

24.3

26.6

0.51-1.00

3.3

2.7

1.8

1.5

1.7

1.1

2.4

3.8

4.3

1.01-1.50

0.5

0.5

0.7

0.2

0.2

0.1

2.4

1.3

0.5

1.51-2.00

0.2

0.1

0.4

0.04

0.05

0.1

0.4

>2.00

0.3

0.3

0.6

0.04

0.5

3.4

0.3

II.

DDT

T-0.03

35.1

50.5

60.1

38.1

53.5

57.5

28.2

23.3

35.6

0.04-0.10

34.3

25.0

18.4

27.9

14.9

15.5

33.3

27.9

14.9

0.11-0.50

25.7

20.8

16.5

31.5

28.3

25.9

23.1

34.9

46.0

0.51-1.00

3.2

2.3

2.6

2.0

2.7

1.1

5.1

7.0

3.4

1.01-1.50

0.6

0.5

0.4

0.2

0.6

10.3

2.3

1.51-2.00

0.3

0.8

>2.00

1.2

0.6

1.1

0.2

4.7

III.

DDE

T-0.03

43.4

45.2

44.4

42.9

40.8

45.3

54.1

43.9

34.2

0.04-0.10

27.3

25.5

26.0

28.7

26.6

27.5

22.9

29.8

25.2

0.11-0.50

25.8

25.3

24.8

26.2

29.9

24.5

23.0

19.3

32.4

0.51-1.00

2.8

3.3

2.2

2.0

2.3

2.0

3.5

7.2

1.01-1.50

0.4

0.4

1.5

0.4

0.3

1.8

1.1

1.51-2.00

0.1

0.2

0.5

0.2

0.3

>2.00

0.1

0.2

0.6

1.8

IV.

TDE

T-0.03

55.1

52.2

56.1

50.3

44.2

43.3

44.0

50.0

48.9

0.04-0.10

25.3

30.9

25.1

29.4

31.0

31.7

8.0

20.6

27.7

0.11-0.50

17.9

14.1

15.3

18.7

22.9

23.3

40.0

11.8

19.1

0.51-1.00

1.2

1.3

1.1

1.4

1.6

1.7

4.0

2.9

4.3

1.01-1.50

0.4

0.9

0.3

0.4

4.0

2.9

1.51-2.00

0.2

0.2

2.9

>2.00

0.2

1.0

1.3

8.8

V. DIELDRIN

T-0.03

37.8

39.2

36.0

37.4

38.3

42.2

51.7

45.9

47.6

0.04-0.10

26.2

25.8

27.8

35.7

28.2

34.7

37.9

43.2

45.2

0.11-0.50

30.2

31.0

33.8

24.8

32.4

23.1

10.3

8.1

4.8

0.51-1.00

5.0

3.7

1.8

1.2

0.8

2.7

1.01-1.50

0.2

0.2

0.2

0.5

2.4

1.51-2.00

0.2

0.1

0.1

0.2

0.3

>2.no

0.5

0.3

0.2

VI. HEPTACHLOR EPOXIDE

T-0.03

30.5

28.7

45.0

37.9

53.8

30.0

57.1

33.3

33.3

0.04-0.10

22.9

26.1

37.9

27.9

22.5

39.0

28.6

16.7

33.3

0.11-0.50

40.5

41.1

17.0

33.4

23.4

31.0

14.3

50.0

33.3

0.51-1.00

4.5

3.2

0.2

0.7

0.3

1.01-1.50

0.8

0.9

1.51-2.00

0.6

>2.00

0.1

VII.

BHC

T-0.03

30.2

71.5

85.5

45.0

60.6

72.0

24.1

23.1

29.7

0.04-0.10

28.1

21.1

9.0

20.2

29.1

10.8

10.3

38.5

24.3

0.11-0.50

34.4

7.4

5.1

33.7

7.3

17.2

58.6

33.3

37.8

0.51-1.00

5.2

1.1

3.0

6.9

5.1

8.1

1.01-1.50

2.1

0.4

1.51-2.00

>2.0U

Vol. 1, No. 3, December, 1967

TABLE 6. — Percent dislrihiilion of residues, by fiscal year and product class, hi difjcrciit quantitative ranges — Continued

"

Percent Distribution of Positive Samples

Range

Fluid Milk

Manufactured Dairy Products

(PPM)
(FAT BASIS)

Domestic

Domestic

Imported

1964

1965

1966

1964

1965

1966

1964

1965

1466

VIII. LINDANE

T-0.03

67.2

66.1

54.4

65.6

84.7

40.0

52.9

28.6

94.1

0.04-0.10

24.5

24.2

10.5

22.9

10.6

20.0

17.6

42.9

0.11-0.50

7.5

9.7

26.3

10.2

4.7

20.0

23.5

28.6

5.9

OJl-1.00

0.8

7.0

1.3

13.3

1.01-1.50

1.8

6.7

1.51-2.00

>2.0C

5.9

IX. ALDRIN

T-0.03

55.6

86.7

81.8

100

100

91.7

96.3

0.04-0.10

44.4

6.7

8.2

8.3

3.7

0.11-O.50

6.7

10.0

0.51-1.00

1.01-1.50

1.51-2.00

>2.00

X. HEPTACHLOR

T-0.03

57.9

55.6

80.0

83.3

100

0.04-0.10

20.9

ll.I

20.0

8.3

100

0.11-0.50

19.4

22.2

8.3

0.51-1.00

11. 1

1.01-1.50

1.51-2.00

>2.00

XI. METHOXYCHLOR

T-0.03

20.0

26.9

38.7

33.3

100

0.04-0.10

68.0

23.1

6.5

33.3

40.0

0.11-0.50

8.0

46.2

25.8

33.3

60.0

0.51-1.00

4.0

16.1

1.01-1.50

3.8

3.2

1.51-2.00

6.5

>2.00

3.2

Discussion

From Table I, it is obvious that within the broad geo-
graphic sections, the samples were reasonably related to
milk production. The relationship was more variable on
an individual State basis. In our opinion, there were
enough samples collected over a wide geographic range
reasonably proportionate to milk production to consider
the findings representative of residues in fluid milk and
other dairy products during the 3-year period.

Over hait of all samples contained one or more residues.
No geographic division was free of residues. The inci-
dence of residues in the East North Central States was
less than half that in most of the country, and the inci-
dence of residues in the adjoining West North Central
States was also significantly lower. The incidence of resi-
dues in the other four ^;eographic divisions — 68.0, 74.2,
74.3, and 77.6% — was not significantly different.

Although almost half of the samples were collected
during FY l'./64, the number of samples collected each

year was considered large enough to yield significant
results.

The data show beyond question that a majority of the
milk and other dairy products marketed in the United
Slates contain detectable quantities of one or more pes-
ticide chemicals. Almost half of the lots examined con-
tained more than one pesticide residue.

The incidence of residues reported was lower in FY 1964
than in FY 1965 or 1966. The closer relationship between
the 1965 and 1966 findings suggests that either there
was a significant increase in residues in milk or the labo-
ratories were more proficient in detecting residues. We
believe the latter is the most logical explanation, because
1964 was the initial year of the program and the first
year that gas chromatography was in general use. In our
opinion, there has been no significant change in the inci-
dence of residues during this period.

The pesticide chemicals shown in Table 3 were found
each year and in each commodity grouping, which is not

I'EsriciDES Monitoring Journal

surprising. The order of frequency varies slightly, but
not by order of magnitude. DDE and TDE are metabo-
lites of DDT, and their presence in milk is to be expected.
The incidence of dieldrin residues in domestic samples
is almost double that found in imported products. The
incidence of BHC residues in domestic products is about
half that found in imported products. The findings on
heptachlor and heptachlor epoxide are noteworthy in
their very low incidence in imported products and the
frequent occurrence of heptachlor epoxide in domestic
milk fat. Heptachlor and aldrin are metabolized and
normally excreted in the milk as heptachlor epoxide and
dieldrin, respectively. The low incidence of heptachlor
and aldrin suggests analytical error, external contamina-
tion after milking, or incomplete conversion to the
epoxide. We are inclined toward the latter two possibili-
ties because of positive findings in several different labo-
ratories in each year and confirmation in the total diet
samples.

The 95% confidence ranges for the averages for each
chemical shown in Table 5 are rather narrow. Specific
attention is directed to the averages of dieldrin and hep-
tachlor epoxide residues. Although each average is equiv-
alent to the average of the individual DDT compounds,
the latter are usually considered in combination which
makes the averages of dieldrin and heptachlor epoxide
about one-third of that resulting from DDT. Considering
the sampling program and procedures, in our opinion,
the averages are reliable indices of the pesticide residue
content of milk and dairy products throughout the United
States during this period. They may be useful as base-
lines to compare future results.

The data were not amenable to consideration of the
various combinations of residues in samples. It is well
known that the DDT metabolites, DDE and TDE, are
most often found in combination with DDT. The rela-
tively high incidence of dieldrin and heptachlor epoxide
suggests that either of these two chemicals may often be
found in milk fat containing the DDT group.

The percent distribution of residue levels is about the
same for each chemical and product when only the
samples containing that chemical are considered as shown
in Table 6. As expected, deviations from the overall
averages become greater as the data are classified in more
detail, but the deviations are not great enough to invali-
date the general statement. These patterns are typical of
residue levels ' i all food classes. A tendency can be ob-
served toward fewer extreme values, above 0.51 ppm, of
the more toxic pesticide chemicals such as dieldrin, hep-
tachlor epoxide, and BHC. It is significant that the per-
cent of values above 0. 1 1 ppm for dieldrin and hepta-
chlor epoxide was substantial and relatively constant,
with the exception of dieldrin in imported products as
noted above.

Table 7 compares the 3-year average values for the 10
most commonly found residues with the averages found
in composites of the dairy portion of 40 total diet sam-
ples examined by FDA from April 1964 through June
1966. The total diet samples are collected at the retail
level representing a different point in the distribution
chain. The results of both investigations are reported on
a fat basis, and since processing techniques used in manu-
facturing dairy products probably do not affect the pesti-
cide residue content of the fat, each should serve as a
check on the reliability of the results.

TABLE 7. — Average levels of pesticide chemicals in dairy
products

(Parts per Million — fat basis)

Pesticide

Objective

Samples
(3-year average)

Total Diet
Samples

(2-YEAR average)

DDE

0.066

0.074

Dieldrin

0.042

0.017

DDT

0.042

0.037

Heptachlor epoxide

0.036

0.010

TDE

0.026

0.013

BHC

0.007

0.008

Lindane

0.004

0.005

Aldrin

0.001

0.001

Heptachlor

0.002

—

Methoxychlor

0.001

0.002

These averages are in remarkably good agreement con-
sidering the extremes in the number of saiiiples repre-
sented. The average levels of dieldrin and heptachlor
epoxide found in the total diet samples are lower than
in the objective samples. These differences exceed the
standard deviation calculated for the total diet samples.
No logical explanation for the lack of agreement in these
two residues is immediately apparent; a search is being
made for the reason. The average level in parts per mil-
lion does not change the order of magnitude for any
pesticide residue.

There are no known approved uses of pesticide chemi-
cals which might result in residues in milk above the
legal tolerance levels. Their presence in milk fat results
from indirect sources, some of which (air, dust, and
drift) are beyond control of the dairyman. Other
sources, such as feed, equipment, and direct application
to animals, are controllable.

Recently, there has been a reduction in the approved
uses and use patterns of some of the more persistent
chlorinated organic pesticides. This reduction in use
would not be reflected in this report because of the time
periods involved.

It is unlikely that the current levels will be reduced or
even remain constant without continued specific attention

Vol. 1, No. 3, December, 1967

by industry and government to eliminating ail controllable
sources and maintaining the residue load from uncon-
trollable sources at a minimum.

No satisfactory system has been designed to identify for
sampling only those lots containing unsanctioned or
excessive residues. While such a system would be the
most effective control, the factors influencing residues
change so rapidly and are so complex and interrelated,
that it is unlikely such control will be practical in the
foreseeable future. There continues to be a need for
information as described in this report concerning the
character and levels of all pesticide residues being con-
sumed.

Significant monitoring programs at production and dis-
tribution centers are capable of identifying problems at
early stages. Corrective measures by government and
industry for consumer protection are most effective dur-
ing these early stages. This type of program serves to
prevent local situations from spreading into national
problems affecting the Nation's health.

Summary and Conclusions
A representative annual sampling of milk and dairy
products marketed during FY 1964, 1965, and 1966
shows that DDE, dicldrin. DDT, heptachlor epo.xide,
TDE, BHC, lindane, aldrin, heptachlor, and methoxy-
chlor account for over 99% of the chlorinated organic
residues in milk and dairy products. TTiese chemicals
were found in each of the 3 years. Twenty-three other
chemicals were found at low levels in 1 or more of the
12,836 samples examined.

Over half of the samples contained residues, and most
of these contained more than one pesticide chemical. The
incidence of residues in the U.S.D.A. East and West
North Central Crop Reporting Divisions was lower than
in other portions of the United States.

A substantial majority, 95%, of the residues found were
below 0.5 ppm on a fat basis, and 71.5% were below
0.1 1 ppm.

No substantial annual variations were noted in these
observations with respect to fluid milk, domestic manu-
factured dairy products, and imported dairy products,
except for dieldrin, heptachlor epoxide, and BHC in
imported dairy products.

The average levels and kinds of pesticide chemicals
found in the objective samples are in good agreement
with the findings on the dairy portion of tola! diet sam-
ples collected at a different point in the distribution
chain, and add a measure of confidence to the total diet
studies as a whole as a broad index to the quantities
of pesticides being consumed in the diet.

The average levels reported are approximately one-tenth
the established tolerance of 1.25 ppm (fat basis) for

DDT, DDE, and TDE residues combined. The average
levels of dieldrin and heptachlor epoxide are approxi-.
mateiy one-tenth of the current administrative determina-
tion of 0.3 ppm (fat basis) for excessive residues for
each chemical. The averages of the remaining five chem-
icals are much lower.

The total pesticide content consists, in a majority of
samples, of a combination of chemicals. The most prob-
able combinations will include one or more members
of the DDT group with dieldrin or heptachlor epoxide.

It is obvious that the total residue content of milk fat
should not be permitted to increase since this is the
source of 13.6% of the total dietary intake (8) of chlori-
nated organic pesticides. The residue pattern indicates
that increases would be accompanied by considerable
loss in economic terms and food value through the
control mechanisms at city, county, State and Federal
levels designed to prevent consumption of dairy products
containing excessive residues.

Even though no major nationwide problem is obvious,
there have been several instances of considerable con-
cern to specific localities during this period. The eflfects
of these incidents were minimized through the coopera-
tive efforts of all sharing the responsibility for an ade-
quate and safe supply of dairy products. Reductions in
the residue content of dairy products can only be made
through a general continued and cooperative effort by
the dairy industry and all agencies of government.

A cknowledgments

Recognition must be given to the chemists, too numerous
to mention as individuals, among the 18 FDA District
Laboratories responsible for these analyses and to R. K.
Dawson, Division of Program Operations, for his as-
sistance in processing the data.

LITERATURE CrFED

(/) Clifford, Paid A. 1957. Pesticide residues in fluid mar-
ket milk. Public Health Rep. 72:729.

(2) Clifford, Paid A., Jonas L. Bassen, and Paid A. Mills.
1959. Chlorinated organic pesticide residues in fluid
milk. Public Health Rep. 74:1109.

13) Association of Official Analytical Chemists. Changes in
official methods of analysis. J. Ass. Offic. Anal. Chem.
49:222 (1966); ihid. 50:210 (1967).

(4) U. S. Department of Health, Education, and Welfare,
Food and Drug Administration. 1963. Revised 1964,
1965. Pesticide analytic;il manual. Vol. 1,

15) Johnson, L. Y. 1965. Colhihorative .study of a method
for multiple chlorinated pesticide residues in fatty
footls. J. Ass. Omc. Ayr. Chem. 48:668.

(6) Welts, Clyde E. December 1967. Validation study of a
niethtxl for pesticide residues in foods and animal
feeds. J. Ass. Oflic. Anal. Chem. (In press).

(7j U. S. Department of Agricidture. 1966 (issued Feb.
1967). Annual statistical summary, milk production
and dairy pnxlucts.

(5) Diignan. R. E. and J. R. Weatherwa.x. 1967. Dietary in-
take of pesticide chemicals. Science 157:1006.

8

Pesticides Monitoring Journal

RESIDUES IN FISH, WILDLIFE
AND ESTUARIES

Chlorinated Pesticide Levels in the Eastern Oyster

(Crassostrea virginica) From Selected Areas of the South

Atlantic and Gulf of Mexico

John C. Bugg, Jr.', James E. Higgins-, and Eric A. Robertson, Jr.''

ABSTRACT

Oysters were collected from estuarinc areas in South
Carolina, Georgia, Florida, Mississippi, Louisiana, and
Texas and analyzed for pesticide residues.
Pesticide levels were determined by the electron capture
gas chromatography method and were confirmed by thin
layer chromatography and dual-column electron capture
gas chromatography.

In general, chlorinated pesticides were either not detected
or were found at relatively low levels in samples collected
from the Atlantic and Gulf Coast areas. Of a total of 133
samples, 94.7% contained I or more pesticides: 89.5%
contained 2 or more; 81.2% contained 3 or more; 63.9%
contained 4 or more; and 31.9% contained 5 or more.
The level of sensitivity for pesticide residues was 0.01 ppm.
Some correlation was found between spraying operations
and pesticide levels in the oysters.

Purpose
The purpose of this report is to present data on the oc-
currence of chlorinated pesticides in oysters in selected
areas of the South Atlantic and Gulf of Mexico as
determined through research conducted at the Gulf
Coast Marine Health Sciences Laboratory on the devel-
opment and evaluation of methodology for the analyses
of chemical contaminants and natural toxins in shell-
fish.

Factual Data
Oysters for this study were obtained from South Caro-
lina, Georgia, Florida, Mississippi, Louisiana, and Texas.
The oysters were either collected directly from oyster-
growing areas by representatives of the State health and
conservation agencies or purchased from oyster dealers
who verified the general locations of the sampling sites.

1 Gulf Coast Marine Health Sciences Laboratory. U. S. Public Health
Service, Dauphin Island, Ala. 36528.

Present Address: Humble Oil & Refining Company, 909 Jefferson
Davis Parkway. New Orleans, La. 7016(1.

- Bureau of Commercial Fisheries Exploratory Fishing and Gear Re-
search Base. LI. S. Fish and Wildlife Service, Pascagoula, Miss. 39567.

3 Gulf Coast Marine Health Sciences Laboratory, U. S. Public Health
Service, Dauphin Island, Ala. 36528.

The oysters were chilled in ice immediately after collec-
tion and then frozen. Frozen shellstock or shucked
oysters were shipped to the Research Laboratory in
insulated containers with dry ice. Immediately upon
arrival, shellstock was shucked and drained of liquor.
Samples not analyzed immediately upon receipt were
stored at — 10 C. No samples were stored over 60 days.
At least a pint of shucked oysters was used for each
sample. The samples were placed in a blender for 5
minutes after which a homogenized 50-g sample was
withdrawn for analysis.

The laboratory methods and techniques used for the
analysis of pesticide residues in oysters were essentially
those compiled by Barry et al. (1). The major deviation
from these methods was the utilization of the "perfo-
rated" basket centrifuge head as described by Robertson
and Tyo (6) for separating oyster meats from the ex-
tracting solvent.

Quantitative determinations of the residues were ini-
tially carried out on a 5% DC-11 column and later on
a mixed column containing equal parts by weight of 10%
DC-200 (12,500 CSTKS) and 15% QF-1 (10,000 CS)
on Gas Chrom Q 60/80 mesh solid support (2) with a
Tritium-parallel plate electron capture detector. The
level of sensitivity was 0.01.

Confirmatory procedures used were thin layer chroma-
tography as described by Kovacs (5), microcoulometry,
and dual differential columns as described by Burke
and Holswade (2). coupled with a Ni'''^ pin cup electron
capture detector and a H ' parallel plate electron capture
detector.

Standard mixtures containing the pesticides were injected
into the gas chromatograph each day before any sample
injection, as well as during the course of injection of
samples for residue determinations. Standards were also
injected after any sample yielding significant pesticide
residues.

Vol. 1, No. 3, December, 1967

Results und Dhctission

The pesticide levels detcclcd in the 133 oyster samples
from South Carolina, Georgia. Florida, Mississippi, Lou-
isiana, and Texas are shown in Appendix 1. Of the total
number of oyster samples, 126 were found to contain
1 or more chlorinated pesticides. For each pesticide, the
number of oyster samples in which the pesticide was
detected and the median, low . and high values of pesticide
concentration in ppm, as taken from Appendix I, are
shown in Table 1 .

Table 2 summarizes the results of analyses of all oyster
samples. The distribution of specific pesticides in posi-
tive samples at different arbitrarily selected residue levels
is shown, as well as the number of samples in which the
specific pesticide was not delccled.

TABLE 1. — Frequency <;/ cliloriiuilal pcsiicidc residues in
oyster samples

I Period of sampling — Feb. 1. 1964 through Aug. 24. 1966|

TABLE 2. — Disirihiiiion of chlorinated pesticides at
different residue levels — all oyster samples

Pesticide

So

1

« <

. X

0 lU

No. Samples

Positive for

Pesticide

Shown

Residue (ppm drained weight)

Median

Low

High

AJdrin

BHC-Lindane

Chlordane

DDD

DDE

PP'-DDT

Dieldrin

Endrin

Hepiactilor

Heptachlor epoxide

Methoxychlor

Toxaphene

Trithion*

133
133
132

81
131
131
115
115
133
133
133
133

55

17

55

20

81

123

117

54

27

12

20

A

6

0

<0.01

0.01

<0.0I

0.02

0.02

0.02

0.01

<0.01

<0.01

<0.01

<0.01

0.08

<0.01
<0.01
<0.01
<0.01
<0.01

<n.oi

<0.01
<0.01
<0.01
<0.01
<0.01
<0.0I

0.03
0.50

0.0 :

0.37

0.12

0.22

0.03

0.07

<0.01

<0.01

<0.01

1.00

Aldrin

BHC-Lindane
Chlordane

DDD
DDE
P.P'-DDT
Dieldrin

Endrin

Heptachlor

Heptachlor epoxide

Methoxychlor
Toxaphene

Triihion «

not less than 95'H- of 1,2,3,4,10,10-hexachloro-I,

4,4a.5.8.8a-hexahydro-l,4-efi(/o-e>:o-5.8-dimethant>=

naphthalene

1.2,3.4.5.A-hexachlorocyclohexane, mixed isomers

l.2,4.5.6.7.8.8-ocla,hlo.-o-3a.4.7.7a-letrahydro-

4.7-melhanoindane

1 . 1 -dichloro-2.2-bis ( p-chlorophenyl) ethane

l,l-dichloro-2.2-bis(/>-chlorophcnyl) ethylene

1,1,1 -trichloro-2.2-bis ( p-chlortipheny I ) 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-1.4-<■«(/o-(^xo-

5,8-dimethanonaphthalenc

1.2. 3.4. 10.1 0-hcxachloro-6,7-epoxy-l . 4,4a.5,6.7,8,8a-

octahydro-l.4-cni/fy-c/i(/o-5.8-dimelhanonaphthaIene

1.4.5,6.7.«.8-hcp:.nchloro-3a,4,7,7a-tetrahydro-

4.7-mcthanoindcne

1,4, 5.6.7.8. 8-hcpl,ichloro-2,3-cpoxy-3a.4,7 .7a-

letrahydro-4,7-methanoindan

1,1.1 -trichloro-2.2-bis? p-methoxyphenyl )cthane

chlorinated camphene conlaininK 67% to 69%

chlorine

.5-1 (p-chlorophenylthio) methyl] 0.0-diethyl

phosphorodithioate

[Period of sampling — Feb. 1, 1964

through Aug. 24, 1966)

No.
Samples
Examined

Residue Levels in ppm Drained Weight

Pesticide

Not
Detected

0.01-
<0.0I 0.05

>0.05

Aldrin

133

116

16

1

0

BHC-Lindane

133

78

27

27

1

Chlordane

132

112

19

1

0

DDD

81

0

2

72

7

DDE

131

8

24

83

16

;».('-DDT

131

14

17

90

10

Dieldrin

115

61

16

38

0

Endrin

115

88

19

7

I

Heptachlor

133

121

12

0

0

Heptachlor epoxide

133

113

20

0

0

Methoxychlor

133

127

6

0

0

Toxaphene

133

127

2

1

3

Triihion®'

55

55

0

0

0

As shown in Table L the high values of pesticide con-
centration in all positive oyster samples ranged from
<0.01 ppm for heptachlor, heptachlor epoxide, and
methoxychlor to 1 .00 ppm for toxaphene. The median
value of pesticide concentration ranged from <0.0I ppm
for aldrin, chlordane, endrin, heptachlor, heptachlor
epoxide, and methoxychlor to 0.08 ppm for toxaphene.
The low value of pesticide concentration was <0,01
ppm for all pesticides.

Endrin and dieldrin, the more toxic of the chlorinated
hydrocarbon pesticides (4). were generally found in low
concentrations. Only 27 of 115 oyster samples were
positive for endrin, the range being from <0.01 to 0.07
ppm. Fifty-four of 115 oyster samples were positive for
dieldrin, the range being from <0.0I to 0.03 ppm. The
chlorinated pesticides found most frequently were p,p-
DDT and two of its metabolites, DDD and DDE, Al-
though these were found at higher concentrations, gen-
erally, than the other chlorinated pjesticides, the levels
of concentration were still relatively low. Maximum
values for p.p-DDT. DDD, and DDE were 0.22 ppm,
0.37 ppm, and 0.12 ppm, respectively.

BHC-lindanc was found in 55 of 133 oyster samples,
with the high, median, and low values being 0.50 ppm,
0.01 ppm, and <0.01 ppm, respectively. Toxaphene was
found in only 6 ot 133 oyster samples, with the high,
median, and low values being 1.00 ppm, O.OS ppm, and
<0,01 ppm, respectively. The high concentrations of
BHC-lindanc and toxaphene were found in the same
oyster sample collected from growing waters affected by
recent application of these pesticides in an adjacent area.
Oyster samples taken from these waters at later dates
showed successively lower levels of these pesticides, with
an eventual decrease to non-detectable levels.

10

Pesticides Monitoring Journal

Aldrin, chlordane, heptachlor, heptachlor epoxide, me-
thoxychlor, and Trithion'® generally were found infre-
quently and in very low concentrations.

The results of the laboratory analyses of all oyster sam-
ples showed that, in general, chlorinated pesticides were
either not detected or were found in relatively low levels
in the positive samples. The ranges of the pesticide levels
in all oyster samples were generally of the same magni-
tude as those found by the U. S. Food and Drug Ad-
ministration in 1964 and 1965 in the analyses of 216
composite samples of 12 major food groups comprising
the American food supply. The amounts of the pesticide
residues found by the Food and Drug Administration
were reported as insignificant from a health stand-
point (3, 7).

Conclusions

In general, chlorinated pesticides were either not de-
tected or were found in relatively low levels in the
oyster samples collected from the South Atlantic and
Gulf of Mexico coastal areas for this study. The data
on chlorinated pesticide concentrations in oysters indi-
cate little or no public health hazard at the present
time. The occasional occurrence of the higher concen-
trations of chlorinated pesticides in oysters as found in
this study, however, indicates that contamination of
shellfish-growing waters with such pesticides does rep-
resent a potential problem that should be kept under
surveillance.

A cknowledgments

The cooperation and assistance of the South Carolina
State Board of Health, Georgia Department of Public

Health, Florida State Board of Health, Mississippi Ma-
rine Conservation Commission, Louisiana State Board
of Health, Louisiana Wild Life and Fisheries Commis-
sion, and Texas State Department of Health in the con-
duct of this study are gratefully acknowledged. The
participation of personnel of these State health and con-
servation agencies included the harvesting and shucking
of shellstock, arranging for the procurement of oyster
samples from dealers, and handling arrangements for
shipment of samples to the Laboratory. These activities
are recognized as a significant contribution to this study
and are deeply appreciated.

LITERATURE CITED

{1} Barry, Helen C, J. G. Hundley, and Loren Y. John-
son. 1965. Pesticide analytical manual. Vol. 1. Revised
ed. Food and Drug Admin. U. S. Dep. Health, Educ,
and Welfare.

(2) Burke, J. A. and W. Holswade. 1966. A gas chroma-
tographic column for pesticide residue analysis: reten-
tion times and response data. J. Ass. Offic. Anal. Chem.
49 (2):374-385.

(3) Duggan, R. E., H. C. Barry, and L. Y. Johnson. 1966.
Pesticide residues in total-diet samples. Science 151
(3706):101-104.

(4) Kenaga, E. E. 1966. Pesticide reference standards of
the Entomological Society of America. Bull. Entomol.
Soc. Amer. 12(2) : 1 17-127.

(5) Kovacs, Martin F., Jr. 1963. Thin-layer chromatogra-
phy for chlorinated pesticide residue analysis. J. Ass.
Offic. Agr. Chem. 46:884-893.

(6) Robertson, E. A. and R. M. Tyo. 1966. Note on im-
proved extraction for chlorinated pesticide residues in
oysters J. Ass. Offic. Anal. Chem. 49(3): 683-684.

(7) U. S. Food and Drug Administration press release.
April 9, 1967.

APPENDIX I. — Distribution of residues of specific chlorinated pesticides, by region
[Period of sampling— Feb. 1, 1964 through Aug. 24, 1966]

South
Carolina

Georgia

Florida

Mississippi

Louisiana

Texas

Total

No. Samples
examined

No. Samples
in which one or
more pesticides
detected

32
29

22
21

44
44

1
1

20
19

14
12

133
126

CHLORINATED PESTICIDES (ppm drained weight)

ALDRIN

No. positive

(1)

(1)

(8)

(0)

(4)

(3)

(17)

Median

_

—

<0.01

—

<0.01

<0.01

<0.01

Low

—

— .

<0.01

—

<0.01

<0.01

<0.01

High

<0.01

<0.01

<0.01

—

<0.01

0.03

0.03

BHC-LINDANE

No. positive

(14)

(5)

(19)

(0)

(9)

(8)

(55)

Median

0.01

<0.01

<0.01

—

<0.OI

0.01

0.01

Low

<0.01

<0.01

<0.01

—

<0.01

<0.01

<0.01

High

0.50

0.01

0.01

—

0.02

0.02

0.50

CHLORDANE

No. positive

(6)

(4)

(8)

(1)

(1)

(0)

(20)

Median

<0.01

<0.01

<0.01

—

—

—

<0.01

Low

<0.01

<0.01

<0.01

—

—

—

<0.01

High

<0.01

<0.01

<0.01

<0.01

0.01

—

0.01

Vol. 1, No. 3, December, 1967

11

APPENDIX I. — Distribution of residues of specific pesticides — Continued
[Period of sampling— Feb. I, 1964 through Aug. 24. 19661

South
Carolina

Georgia

Florida

Mississippi

Louisiana

Texas

CHLORINATED PESTICIDES (ppm drained weight) (Continued)

Total

DDD'

No. positive

111)

(15)

(40)

(1)

(10)

(4)

(81)

Median

0.02

0.02

0.02

—

0.01

0.02

0.02

Low

0.01

0.01

<0.01

—

<0.0I

0.01

<0,01

High

n.05

0.04

0.37

0.02

0.07

0.05

0.37

DDE

No. positive

(29)

(19)

(44)

(1)

(18)

(12)

(123)

Median

., M-.

0.02

0.02

—

-

O.OI

0.02

Low

<0.01

<0.01

—

<0.01

<0.01

<0.01

High

0.04

0.12

0.02

0.02

0.04

0.12

p.p-DDT

No. positive

125)

(19)

(44)

(1)

(16)

(12)

(117)

Median

O.OI

0.02

0.02

—

0.01

0.02

0.02

Low

<0.01

<0.01

<0.01

—

<0.0I

<0.01

<0.01

High

O.OJ

0.0.1

0.22

0.02

0.06

0.07

0.22

DIELDRIN

No. positive

(11)

(12)

(16)

(1)

(9)

(5)

(54)

Median

0.01

0.01

0.01

—

0.01

0.01

0.01

Low

<0.01

<0.01

<0.01

—

<0.01

0.01

<0.01

High

0.02

0.02

0.03

<0.01

0.01

0.03

0.03

ENDRIN

No. positive

(1)

(6)

(7)

(1)

(8)

(4)

(27)

Median

<0.01

0.01

0.01

—

^

2

<0.01

Low

<0.01

<0.01

<0.0I

—

<0.01

<0.01

<0.0I

High

<0.01

0.07

<0.01

<0.01

0.02

0.02

0.07

HEPTACHLOR

No. positive

(1)

(1)

(5)

(0)

(2)

(3)

(12)

Median

<0.01

—

<0.01

—

<0.01

<0.01

<0.01

Low

<0.01

—

<0.01

—

<0.01

<0.01

<0.01

High

<0.01

<0.01

<0.01

—

<0.01

<0.0I

<0.01

HEPTACHLOR

EPOXIDE

No. positive

(2)

(0)

(9)

(0)

(7)

(2)

(20)

Median

<0.01

—

<0.01

—

<0.01

<0.01

<0.01

Low

<0.01

—

<0.01

—

<0.0I

<0.01

<0.01

High

<0.01

—

<0.01

—

<0.01

<0.01

<0.01

METHOXYCHLOR

No. positive

(2)

(1)

(2)

(0)

(1)

(0)

(6)

Median

<0.01

—

<0.01

—

—

—

<0.01

Low

<0.01

—

<0.01

—

—

—

<0.01

High

<0.01

<0.01

<0.01

—

<0.01

—

<0.01

TOXAPHENE

No. positive

(6)

(0)

(0)

(0)

(0)

(0)

(6)

Median

0.08

—

—

—

0.08

Low

<0.01

—

—

<0.01

High

1.00

—

—

—

—

—

1.00

TRITHION*

No. positive

(0)

(0)

(0)

(0)

(0)

(0)

(0)

Median

—

—

—

—

—

Low

—

High

—

—

—

—

—

—

—

' DDD not included in routine analysis until October 1964.
- Indeterminate.

12

PE.sTiriDES Monitoring Journal

Galveston Bay Pesticide Study — Water and Oyster Samples
Analyzed for Pesticide Residues Following Mosquito Control Program

Victor L. Casper^

ABSTRACT

The purpose of tliis study was to determine the effect of in-
creased pesticide applications in the Houston area on sliellfish
and shellfish-growing waters of Galveston Bay.
The study was conducted during the fall of J 964 following a
large-scale mosquito control program in the Houston area.
Water and oyster samples were collected in September and
October 1964, during and after the mosquito control opera-
tions. Oyster samples collected in this study were compared
to samples collected from April to July 1964, prior to the
mosquito operations.

Analyses included determination of levels of BHC-lindane,
DDE, DDT, dieldrin, endrin, heptachlor, aldrin, chlordane,
heptachlor epoxide, methoxychlor, toxaphene, and Trithi-
on'^ . Pesticide levels were determined by the use of electron
capture gas-liquid chroniatography, with thin layer chroma-
tography for confirmation.

Pesticide levels in both water and oysters were low at all
times. The data indicate little or no increase in levels due
to the control program in Houston.

Purpose

The purpose of this study was to determine the effect
of increased pesticide applications in the Houston area
on shellfish and shellfish-growing waters of Galveston
Bay.

Factual Data

Following an outbreak of equine encephalitis in the
Houston area during the summer of 1964, a large-scale
mosquito control program was begun which utilized con-
siderable quantities of pesticides, especially malathion,
DDT, and BHC. Actual spraying operations began the
third week of August. With the increased use of pesti-
cides in the Houston area, the Texas State Department
of Health became concerned over potential pesticide
contamination of shellfish-growing waters in Galveston
Bay. On September 2, officials of the Texas State De-
partment of Health requested the PHS Regional Office
and, in turn, the Gulf Coast Marine Health Sciences

1 Gulf Coast Marine Health Sciences Laboratory, U. S. Public Health
Service, Dauphin Island, Ala. 36528.

Laboratory to provide assistance in laboratory analyses
of pesticides in water and oysters.

Following discussions between representatives of the
Texas State Department of Health and the Laboratory,
a sampling program was established for the collection
of water and oyster samples in Galveston Bay at loca-
tions shown in Figure 1 . Sampling activities were begun
on September 3 and completed on October 6. 1964.

Water and oyster samples were collected by personnel
of the Texas State Department of Health. Water sam-
ples were collected in 1 -gallon chemically clean glass
jugs at nine stations for 5 consecutive weeks. Each
week water samples were shipped unrefrigerated to the
Gulf Coast Marine Health Sciences Laboratory and

FIGURE 1. — Pesticide sampling stations in Galveston Bay

OYSTER REEFS
A Todd's Dump
B Hanna's Reef
C Scoffs Reef
D Redfish Bar

O Water Sampling
Station

• Oyster Sampling
Station

Vol. 1, No. 3, December, 1967

13

TABLE 1. — Summary of oyster analyses
[ND = Not detected]

Date of
samplino

Pesticides in ppm wet drained weighti

bAMPLINu InilNi

BHC-

DDE

DDT

DIELDRIN

ENDRIN

Heptachlor

Aldrin

Others!

LlNDANE

EAST GALVESTON BAY

4/23/64

ND

ND

ND

ND

ND

ND

ND

ND

(Lease 357-A)

EAST GALVESTON BAY

5/11/64

ND

ND

ND

ND

ND

ND

ND

ND

(Miller's Reef)

EAST GALVESTON BAY

6/04/64

0.01

oja

0.01

ND

ND

ND

ND

ND

EAST GALVESTON BAY

6/24/64

0.01

0.01

0.01

ND

ND

ND

ND

ND

EAST GALVESTON BAY

7/07/64

ND

0.05

0.073

0.01

0.02

ND

0.03

ND

(Blumc's Reef)

EAST GALVESTON BAY

7/18/64

ND

0.05

O.is

0.01

0.01

ND

ND

ND

(Blumc's Reef)

GALVESTON BAY

9/03/64

<0.01

0.01

0.01

<0.01

<0.01

ND

ND

ND

(Todd's Dump)

GALVESTON BAY

9/09/64

<0.0I

<0.01

0.01

<0.01

<0.01

<0.01

ND

ND

(Hanna's Reef)

GALVESTON BAY

9/14/64

<0.01

<0.01

0.01

<0.01

<0.01

ND

ND

ND

(Scolt's Reef)

GALVESTON BAY

9/14/64

<0.01

0.02

0.02

<0.01

<0.01

<0.01

ND

ND

(Rcdfish Bar)

1 Analysis by gas chromatography with electron capture.

- Includes chlordane. heptachlor epoxide, methoxychlor. toxaphene, and Trithion®.

3 p,p'-DDT.

refrigerated until ready for analysis. Following collec-
tion, the oysters were shucked into 1 -gallon cans aboard
the collecting vessel and packed in ice. Upon return to
shore, the oysters were frozen, packed in dry ice, and
shipped to the Laboratory. After arrival, the oysters
were kept frozen until ready for analysis.
Due to difficulty in obtaining oysters during the closed
season, only four oyster samples were collected during
the study period. However, six oyster samples collected
between April 23 and July 18 had been analyzed for
chlorinated pesticides in connection with studies at the
Laboratory on development of analytical techniques.

The oysters were prepared for analysis by homogenizing
1 pint of the shucked oysters in a blender for 5 minutes
after which a 50-g aliquot was withdrawn for analysis.
One liter of water was used for the water analysis.

Laboratory examinations were made for residues of the
following chlorinated hydrocarbon pesticides: aldrin.
BHC-lindane, chlordane. DDE. DDT, endrin. dieldrin,
heptachlor. heptachlor epoxide, methoxychlor. and Tri-
thion'*'. Electron capture gas-liquid chromatography was
used for both qualitative and quantitative determinations
and thin layer chromatography for confirmation of
results of oyster analyses. The methods and procedures
were those used by the Food and Drug Administration
(I), with confirmatory procedures by Kovacs (2).

Since malathion, in addition to chlorinated pesticides,
was used for mosquito control operations, studies were
initiated to develop analytical techniques for detection
of organo-phosphate residues in water samples. Water
samples were extracted as described by Tyo (3) for

organo-phosphate residues, and laboratory analyses were
conducted using electron capture gas-liquid chromatog-
raphy.

Standard mixtures containing the pesticide were injected
each day prior to injection of samples, as well as during
the course of injection of samples. Additional standards
were injected after any samples having a significant
pesticide residue.

Findings of chlorinated pesticides were reported at levels
of 0.01 ppm and 0.001 ppm or greater for oysters and
water, respectively. The sensitivity limit for organo-
phosphates was 0.008 ppm. All positive samples having
pesticide concentrations below these levels were re-
ported as "less than." Results of samples in which pesti-
cides were not found are reported as "not detected."

Results and Discussion

Oyster Analyses — The results of pesticide analyses of
six oyster samples collected prior to August 1964 and
four samples collected between September 3 and 14
from Galveston Bay are shown in Table 1 .
Results of analyses of the four oyster samples collected
during and following the mosquito control operations
showed concentrations of DDE and DDT between <0.01
and 0.02 ppm. All four samples contained trace amounts
(<0.01 ppm) of BHC-lindane, dieldrin, and endrin,
while heptachlor was found in two of four samples at
<0.0I ppm. Those chlorinated pesticides not detected
were aldrin. chlordane. heptachlor epoxide, methoxy-
chlor. toxaphene. and Trithion"". Of the six oyster sam-
ples collected during the methodology study earlier in
the year, four were positive for one or more chlorinated

14

Pesticides Monitoring Journal

TABLE 2. — Summary of wciler cmalyses
[ND = Not detected]

Date of

sampling

Pesticides in ppmI

Sampling point

BHC-

LlNDANE

DDE

DDT

Heptachlor

Heptachlor

EPOXIDE

Methoxy-
chlor

Chlordane

Others^

Organo-
phosphates

GALVESTON BAY
(Todd's Dump)

9/03/64

<0.001

ND

ND

ND

ND

ND

ND

ND

Not
examined^

TRINITY BAY

(Station 116)

9/08/64

ND

ND

ND

ND

ND

ND

ND

ND

Do.

EAST BAY
(Hanna's Reef)

9/09/64

<0.0Ol

<0.001

<0.001

ND

ND

ND

ND

ND

Do.

TRINITY BAY

(Station 108)

9/14/64

ND

<0.001

ND

<0.001

ND

ND

ND

ND

Do.

GALVESTON BAY

(Station 226)

9/14/64

ND

<0.001

ND

<0.001

ND

ND

ND

ND

Do.

GALVESTON BAY
(Scott's Reef)

9/22/64

<0.001

ND

ND

ND

ND

<0.001

ND

ND

ND

TRINITY BAY

(Station 61)

9/22/64

ND

ND

ND

ND

<O.0OI

ND

ND

ND

ND

WEST BAY
(Station A^9)

10/06/64

ND

ND

ND

<0.00I

ND

ND

<0.001

ND

ND

WEST BAY
(Station A-59)

10/06/64

ND

ND

<0.001

<0.001

ND

ND

<0.001

ND

ND

t Analysis by gas chromatography with electron capture.
~ Includes aldrin, dieldrin, endrin, toxaphene. and Trithion®.
3 Analytical technique under development.

pesticides. Four samples were positive for DDE and
DDT with ranges of 0.01-0.05 ppm and 0.01-0.1 ppm,
respectively. Concentrations of BHC-lindane, dieldrin,
endrin, and aldrin were each detected in one or two of
the six samples and ranged from 0.01 to 0.03 ppm. Those
pesticides not detected were chlordane, heptachlor epox-
ide, methoxychlor, toxaphene, and Trithion"^.

Water Analyses — Results of the analyses of nine water
samples collected during and following the mosquito
control operations are shown in Table 2.

Eight of nine samples were positive for one or more
chlorinated pesticides, and all pesticide concentrations

The chemical names of compounds mentioned in this paper are:
BHC-Lindane 1,2,3.4,5,6-hexachIorocycIohexane, mixed isomers

DDE l.l-dichloro-2.2-bis(p-chlorophenyl)ethylene

DDT 1, 1, l-trichloro-2,2-bis(p-chIorophenyl) ethane

Dieldrin not less than 85% of 1.2..3,4.10.10-hexachIoro-6,

7-epoxy-l ,4,4a,5,6,7.8,8a-octahydro-l ,4-en(/o-?A:o-

5,8-dimethanonaphthalene
Endrin l,2,3,4,10,10-hexachloro-6.7-epoxy-l,4,4a, 5,6.7,8, 8a-

octahydro-1.4-ertrfrj-£';ido-5,8-dimethanonaphthalene

Heptachlor 1 ,4,5,6,7, 8, 8-heptachloro-3a,4,7,7a-tetrahydro-

4,7-methanoindene
Aldrin not less than 95% of 1,2.3.4,10,10-hexachloro-l,

4,4a,5,8.8a-hexahydro-l,4-e«rfo-e.vo-5,8-dimethano^

naphthalene
Chlordane 1 ,2.4,5,6,7,8,8-octachloro-3a,4,7,7a-tetrahydro-

4.7-methanoindane
Heptachlor epoxide l,4,5.6,7,8,8-heptachloro-2,3-epoxy-3a,4,7.7a-

tetrahydro-4,7-methanoindan
Methoxychlor l,l.l-trichloro-2,2-bis(p-melhoxyphenyl) ethane

Toxaphene chlorinated camphene containing 67% to 69%

chlorine
Trithion® 5-| (p-chlorophenylthio)methyl I 0,0-diethyl

phosphorodithioate
Malathion diethyl mercaptosuccinate, 5-ester with 0,(?-dimethyl

phosphorodithioate

were <0.00l ppm. No sampling station was found to
be positive for more than three pesticides, and no single
pesticide appeared dominant. Those chlorinated pesti-
cides not detected were aldrin, dieldrin, endrin, toxa-
phene, and Trithion's. No organo-phosphates were de-
tected within the limits of the analytical techniques used
in this study.

Conclusions

Pesticide levels in both water and oysters were low at
all times, even during and after the period of intense
mosquito control activity. There was no indication of
elevated pesticide levels because of the mosquito control
operations. In fact, the limited data indicate higher levels
of DDE and DDT in oysters prior to the beginning of
the mosquito control operations.

Acknowledgments
Laboratory analyses of oyster and water samples were
performed by Mr. Robert M. Tyo, Mr, E. A. Robertson,
Jr., Mr. Lavern C. Walters, and Mr. James E. Higgins.
The author would like to express his grateful apprecia-
tion to the Texas State Department of Health for its
assistance in collection and shipment of samples.

LITERATURE CITED

(/) Barry, Helen C. and J. G. Hundley. 1963. Pesticide
analytical manual. Vol. \. Revised ed. Food and Drug
Admin., U. S. Dep. Health, Educ, and Welfare.

(2) Kovacs, Martin F., Jr. 1963. Thin-layer chromatog-
raphy for chlorinated pesticide residue analysis. J. Ass.
Offic. Agr. Chem. 46:884-893.

(3) Tyo, R. M. 1965. Extraction of the organo-phosphate
insecticides from water samples for residue analysis.
Tech. Rep. Gulf Coast Shellfish Sanit. Res. Center.

Vol. I, No. 3, December, 1967

15

Investigation of Effects of Large-Scale Applications
of 2,4-D on Aquatic Fauna and Water Quality'

Gordon E. Smith and Billy G. Isom

ABSTRACT

In 1966, the Tennessee Valley Authority tip plied S88 tons
of 20% 2,4-D, butoxyelhanol ester, granular herbicide to
8.000 acres of Eurasian wtitermilfoil growths in seven res-
ervoirs, at rales varying from 40 to 100 Ih of 2,4-D acid
equivalent per acre. Laboratory analyses showed little up-
take of 2.4-D by fish, but some by mu.'/sels. All mud sam-
ples contained 2.4-D. in varying concentrations. Eight of
nine water treatment plants sampled showed 2,4-D con-
centrations of less than I ppb. The ninth was the only
plant at which 2.4-D was applied directly above the water
intake supply, and its highest concentrations were 2 and
I ppb. respectively. Extensive pre- and post-monitoring data
indicate that high application rates of 2.4-D for watermil-
foil control on TV A reservoirs have not produced adverse
effects on aquatic fauna or water quality.

Introduction

Eurasian watermilfoil {Myriophylhtfn spicatum L.), a
submersed aquatic plant, was first introduced into a
TVA reservoir about 1953. By 1966. it had spread to
seven TVA reservoirs and was posing serious threats to
mosquito control, recreation, navigation, and many other
water uses (1). Plans were made in 1966 to use 20%
2,4-D. butoxyelhanol ester (BEE), granular herbicide to
treat all known colonies of watermilfoil in Melton Hill,
Watts Bar, Chickamauga. Hales Bar. Guntersville,
Wheeler, and Wilson Reservoirs. Some 8.000 acres were
scheduled for treatment at rates from approximately 40
to 100 lb of 2.4-D acid equivalent per acre. Earlier
work had shown watermilfoil to be highly susceptible
to 2,4-D, while other aquatic organisms were relatively
unaffected by it.

From March through December 1966, TVA applied
888 tons of 20% 2,4-D granular herbicide, or 355,200
lb of 2.4-D acid equivalent, to about 8.000 acres of
watermilfoil growing in seven reservoirs from Melton
Hill in cast Tennessee to Wilson Dam in north Alabama.

' Division of Health and Safely. Tennessee Valley Authority, Muscle
Shoals. Ala. 33660.

These reservoirs are spread over a main-channel distance
of 352 river miles. They have 4,000 miles of shoreline,
237,000 water-surface acres, and hold about 4,600,000
acre-feet of water at normal full-pool elevation.

On Watts Bar and Melton Hill Reservoirs, 617 acres of
hard-to-kill milfoil were treated at the rate of approxi-
mately 100 lb of 2,4-D per acre (Watts Bar, 578 acres;
Melton Hill. 39 acres). Previous treatments at a lower
rate were unsuccessful in controlling this aquatic plant.
The remaining 7,383 acres in the other five reservoirs
were treated at the rate of approximately 40 lb of 2,4-D
per acre, with Hales Bar and Guntersville receiving
most of the treatment. These rates of application are two
to five times greater than those used in previous years.

To collect the best possible data on the toxicity of 2,4-D,
outside agencies interested in this problem were urged
to join TVA in planning and carrying out extensive and
intensive monitoring of the watermilfoil control program.
Those invited to participate in the cooperative research
project were representatives of the U. S. Department of
the Interior (Fish and Wildlife Service and Federal Water
Pollution Control Administration); U. S. Department of
Agriculture (Agricultural Research Service): U. S. De-
partment of Health, Education, and Welfare (Public
Health Service); and State agencies of Tennessee and
Alabama. Within TVA, the Reservoir Ecology Branch,
Water Quality Branch, Fish and Wildlife Branch, and
Public Health Engineering StalT joined forces in the
study.

Before, during, and after the 1966 large-scale applica-
tions of 2,4-D, vast amounts of monitoring data were
collected. The purpose of this paper is to summarize
some of these data.

Effects of 2,4-D on Insectary Mosquito Larvae
Prior to field monitoring, a simple laboratory experiment
was conducted, first, to measure the toxicity of 2,4-D to
confined mosquito larvae, and, secondly, to determine
if exposure of immature mosquito stages to exceptionally

16

Pesticides Monitoring Journal

high concentrations of this herbicide would affect repro-
ductive capabilities of the adults. A concentrated solution
(69.3% acid equivalent) of butoxyetlianol ester of 2,4-D
was mixed with ethanol alcohol for use in this experi-
ment in the ratio of 0.45 ml of 2,4-D to 99.55 ml of
alcohol. Six plastic cans were used, each containitig 1
cu ft of water and about equal numbers of third and
fourth instar Anopheles quadrimaculatus Say mosquito
larvae. Five pans of larvae were treated with the 2,4-D-
alcohol solution at the rate of 100 ppm, and one pan
was left for control. Each pan contained about 2,000
larvae by sample count and visual estimate.

About two-thirds more of the larvae in the control pan
reached the pupal stage than did larvae in each of the
five treated pans. Thus, an apparently consistent degree
of mortality occurred in the treated larvae; however,
some of them remained alive in the 100-ppm 2,4-D solu-
tion for as long as 8 days before emerging as adults.
From the larvae and pupae which persisted in the five
pans of 2,4-D-alcohol solution, 85.7% emerged to the
adult stage, while only 79.3% of the pupae from the
untreated larvae in the control pan became adult mos-
quitoes.

Two colonies of mosquitoes were established and main-
tained in separate insectary cages — one from the treated
group and one from the untreated group. Both were car-
ried to the F.. generation. Adult mosquitoes mated, took
blood, and oviposited viable eggs. No difference in hardi-
ness or reproductive ability could be detected.

The rate of treatment in this experiment was, of course,
many times the maximum level attainable in the field
even immediately following treatment. This test showed
that some mosquito larvae can survive 2,4-D exposure
even at the staggering rate of 100 ppm.

Analytical Results of 2,4-D Monitoring
at Water Treatment Plants

The 2,4-D residue in water was monitored at nine water
treatment plants along the Tennessee River system by
use of carbon filters (Davidson. C. M. and K. L. Shalibo.
1967. Analytical results of 2,4-D monitoring at water
plants. Unpublished TVA report). Special filter units
were provided by the Federal Water Pollution Control
Administration, Athens, Ga., and 72 samples were taken
for analysis. Samples were collected at each station prior
to 2,4-D application to determine whether 2,4-D was
already present in the water. Continuous monitoring
began at Watts Bar Dam immediately after the first
2,4-D treatments started. As the treatment operation
moved downstream, monitoring began at other stations
and continued approximately 2 to 3 weeks at each
station following chemical application.

The flow rate through each carbon unit was determined
each hour to assure an accurate record of the volume of

water filtered. Raw water was passed through the carbon
filter unit only when the water plant was operating. After
approximately 500 gallons of water had passed through
the carbon unit, the unit was removed and replaced with
a unit containing clean carbon. The used carbon was
removed from its pyrex container and dried — adsorbed
2,4-D was extracted with ethanol at the TVA Water
Quality Laboratory in Chattanooga. The extracted sam-
ple was then shipped to the Southeast Water Laboratory
of the Federal Water Pollution Control Administration
in Athens, Ga., and analyzed for 2,4-D using electron
capture and microcoulometric gas chromatography. The
weight of 2,4-D in the extracted sample divided by the
volume of water filtered equaled the concentration of
2,4-D expressed in micrograms per liter or parts per
billion.

No recovery studies were performed in this study to de-
termine the rate of adsorption on and desorption from
activated charcoal. Accuracy and precision figures for
this method can be found in the JAOAC 45:367 (1962).
Sensitivity for the liquid-liquid extractions using a 250-ml
aliquot of sample is 10 ppb acid equivalent and less than
1 ppb by carbon adsorption. However, work was done
in the FWPCA Southeast Water Laboratory to determine
2,4-D degradation after sample collection. Values re-
ported indicate minimums present.

Results showed that samples from eight water treatment
plants contained concentrations of less than 1 ppb for
both the butoxyethanol ester of 2,4-D and 2,4-D acid.
The highest concentrations of 2,4-D were found in raw
water samples collected at Scottsboro, Ala., following
application. Prior to the 2,4-D application, the Scotts-
boro water intake had been clogged by watermilfoil, and
2,4-D at the rate of 40 lb per acre was applied directly
over the water intake supply. The raw water sample
collected during the 3 days immediately following appli-
cation contained 2 jxg/ 1 , and the raw water sample col-
lected 4 to 9 days after herbicidal treatment contained
1 j_i.g/ 1 of herbicide. Finished water samples from the
treatment plant contained <1 /.ig/ 1 or no herbicide.

Laboratory detection of the higher levels of 2,4-D at
the heavily treated Scottsboro plant lends support to the
accuracy of the other tests.

Effects of 2,4-D Upon Aquatic Organisms
and Its Persistence in Mud and Water

In 1966 and 1967, the effects of butoxyethanol ester of
2,4-D on aquatic organisms were studied at 21 stations
in a 5-acre embayment (Gordon Branch) on Watts Bar
Reservoir. Water samples were taken from five stations.
In a 275-acre slough above Comer Bridge in Gunters-
ville Reservoir, six stations were selected for study. The
Watts Bar test site was treated with a 20% granular ma-
terial at the rate of 100 lb of 2,4-D acid equivalent per

Vol. 1, No. 3, December, 1967

17

acre, and Guntersville Reservoir was treated at the rate
of 40 lb per acre.

The toxic effect of 2,4-D was evaluated hy sampling the
benthic invertebrate communities of both reservoirs be-
fore treatment and at least twice after treatment. Residue
analyses of water, fish, plants, mussels, and sediment
were used to study dilTusion, accumulation, translocation,
and/ or degradation of 2,4-D.

Since I960, TVA. in cooperation with State fish and
wildlife representatives, has routinely surveyed treatment
areas for dead or distressed native fish following herbi-
cidal applications for milfoil control. None have been
found. This survey has consisted of visual inspection of
water and shoreline before and after treatments. In the
1966-67 studies, TVA fishery biologists, in cooperation
with State representatives, set up concurrent monitoring
stations on Watts Bar and Guntersville to observe the
effects of treatment at 40 and 100 lb per acre on both
caged and free-swimming native fish. It was observed
that, in general, both concentrations appeared to result
in some movement of lake fish out of the treated area.
Again, no mortality of native fish in the treated areas
of the lakes was found.

In the Watts Bar area, percent mortality of caged fish
compared before and after treatment differed signifi-
cantly at the 5''r level of probability. Comparing control
and test cages, percent mortality was significantly higher
for test cages at 72 and 96 hours of exposure. (Chance,
C. J. 1967. Monitoring tests of fish mortaUty in connec-
tion with TVA reservoir milfoil treatment. 1966. Unpub-
lished TV A report). However, we do not believe that this
mortality was due to 2,4-D alone since the concentration
in the water was much below the median tolerance limit
for fish.

SAMPLING PROCEDURES

Benthic fauna and mud samples were collected with a
0.9-sq-ft Petersen dredge. Fish were collected with gill
nets for 2,4-D analysis. Bluegills (Lepomis macrochirus)
were placed in test areas before treatment and removed
at various time intervals after treatment for analysis for
2.4-D, Mussels, primarily EUipiio crassidcns. were used
to monitor uptake of 2,4-D by these invertebrates. Mus-
sels were placed in the test areas prior to treatment, and
some specimens were removed at various times after
treatment and analyzed for 2.4-D.

Of the six stations selected for study on Guntersville
Reservoir, three were established on the quiet overbank
area with little or no current, and three on the old river
channel where there was usually a current. Stations re-
ferred to as "in" were on the overbank in the embay-
ment. Stations referred to as "out" were on the margin
of the channel nearest the cmbayment. A seventh station
referred to as "control" in Table I, which shows results

of 2,4-D analyses, received an unplanned application of
2.4-D and, as a consequence, cannot be considered as a
control. The prestudy data were used as the control fc
analysis of variance.

Fifty assorted frozen samples of plants, animal tissues,
and mud from Watts Bar and Guntersville Reservoirs
were analyzed for 2,4-D by the C. W. England Labora-
tories. Washington, D. C. Tissues of fish, mussels, and
plants were each ground in a high-speed blender; then
samples were removed for analysis. Sensitivity of the
chemical test was 0.14 mg/kg as BEE; however, recov-
ery of known standards when added to our samples was
from 52% to 72%. (Wimsatt, J. C. 2.4-D determination
in shellfish iisini; GLC. Unpublished method — Nat. Cen-
ter Urban and Ind. Health. U. S. Public Health Serv.,
Cincinnati. Ohio). Thus, there would be a tendency
toward underestimating actual concentrations present
rather than overestimating. It should be noted that
analytical results on one sample each of mud and mussel
showing higher concentrations of 2.4-D by this proce-
dure were confirmed by paper chromatography. Values
are reported on wet weight basis (Table 1).

2,4-D ANALYSIS

In Watts Bar Reservoir, watermilfoil samples collected
after a 24-hour exposure showed 2,4-D concentrations
(BEE) up to 8.26 mg/kg. This figure apparently repre-
sents the result of active uptake and translocation of
2,4-D, since 1 hour after treatment only 37 ^iig/ I BEE
was found in water samples (Table 2). Less than 1 /ig/1
BEE was present after 8 hours. However, significant
concentrations of 2,4-D were found accumulated in mud
samples, with detections ranging from 0.14 mg/kg to
58.8 mg/kg BEE.

Two samples of mussels, held in cages for 96 hours in
the treated area, showed concentrations of 0.38 mg/kg
and 0.70 mg/kg BEE. The ratio of 2,4-D content in
water to that in mussels indicates that they concentrated
2.4-D. Eight fish samples (4 species) were negative for
2.4-D. One sample of bluegills (Lepomis macrochirus).
collected 50 days after the area was treated, contained
0.15 mg/kg BEE (Table I).

Mussels and clams held in cages for 30 days in the
Watts Bar test area showed no ill effects from their in-
carceration in this environment which had received a
massive dose of 2.4-D.

Three fish samples from Guntersville were negative for
2.4-D. Mussels exposed to treated water for 24 and 72
hours on the overbank were positive for 2,4-D. Concen-
tration of BEE in fish ranged from 0.24 mg/kg to 1.12
mg/kg BEE. Mussels held at stations near the channel
during the same period were negative for 2,4-D. Mussels
exposed 144 hours in Guntersville were negative for
2.4-D; however, one sample exposed 42 days showed
0.20 mg/kg BEE.

18

PESTicroES Monitoring Journal

TABLE 1. — 2,4-D Analyses — Watts Bar and Gunlersvillc Reservoirs

Sample
No.

Date
Collected

Hours/Days

After
Treatment

Material

MO/11
OR MO/kO

BEE

Stations

Species

Watts Bar Reservoir

19

Prestudy

Control

Fish

< 0.14

Control

Lepomis macrochirus

25

3-20-66

24 hours

Watermilfoil

< 0.14

Gordon Branch

Myriophyllum spicatum

18

3-20-66

24 hours

Watermiltoil

8.26

do.

Myriophyllum spicatum

16

3-20-66

24 hours

Watermilfoil

3.36

do.

Myriophyltum spicatum

21

3-22-66

72 hours

Fish

< 0.14

do.

Lepomis macrochirus

26

3-23-66

96 hours

Mud

56.0

do.

27

3-23-66

96 hours

Mussel

0.38

do.

Assorted mussels

22

3-23-66

96 hours

Mud

2.8

do.

23

3-23-66

96 hours

Mud

0.95

do.

IS

3-23-66

96 hours

Fish

< 0.14

do.

Lepomis macrochirus

8

3-23-66

96 hours

Mussel

0.70

do.

Elliptio crassidens

32

4-13-66

24 days

Mud

35.0

do.

34

4-13-66

24 days

Mud

0.15

do.

33

5-24-66

35 days

Mud

0.14

do.

6

5-25-66

50 days

Fish

< 0.14

do.

Lepomis macrochirus

10

5-25-66

50 days

Fish

< 0.14

do.

Lepomis macrochirus

48

5-25-66

50 days

Fish

< 0.14

do.

Ictahirus pimctatus

49

5-25-66

50 days

Fish

< 0.14

do.

Stizostedion canadense

50

5-25-66

50 days

Fish

0.15

do.

Lepomis macrochirus

47

1-17-67

10 months

Fish

< 0.14

do.

Pomolobus chrysochloris

43

1-17-67

10 months

Mud

0.24

do.

44

1-17-67

10 months

Mud

0.91

do.

45

1-17-67

10 months

Mud

0.28

do.

46

1-17-67

10 months

Mud

58.8

do.

Guntersville Reservoir

17

Prestudy

Control

Mussel

< 0.14

Control

Elliptio crassidens

2

Prestudy

Control

Asiatic Clams

< 0.14

—

Corbicula manillensis

5

May 1966

Control

Mud

0.14

—

11

4-06-66

24 hours

Mussel

0.25

In-1

Elliptio crassidens

13

4-06-66

24 hours

Mussel

0.24

"Control"

Elliptio crassidens

14

4-06-66

24 hours

Mussel

< 0.14

Out-1

Elliptio crassidens

30

4-06-66

24 hours

Mussel

1.12

In-3

Elliptio crassidens

12

4-08-66

72 hours

Mussel

0.18

"Control"

Elliptio crassidens

1

4-08-66

72 hours

Mussel

0.30

In-1

Elliptio crassidens

40

4-08-66

72 hours

Mussel

0.98

In-3

Elliptio crassidens

41

4-08-66

72 hours

Mussel

1.0

In-2

Elliptio crassidens

4

4-11-66

144 hours

Mussel

< 0.14

In-1

Elliptio crassidens

24

4-11-66

144 hours

Mussel

< 0.14

In-3

Elliptio crassidens

39

4-21-66

15 days

Fish

< 0.14

In-2

Ictalurus furcatus

28

4-11-66

144 hours

Mussel

< 0.14

Out-3

Elliptio crassidens

20

4-11-66

144 hours

Fish

< 0.14

Out-1

Lepomis macrochirus

7

5-17-66

42 days

Mud

< 0.14

Out-1

29

5-17-66

42 days

Mud

33.6

In-3

31

5-17-66

42 days

Mud

0.14

Out-2

3

5-17-66

42 days

Mussel

< 0.14

In-1

Elliptio crassidens

9

5-17-66

42 days

Mussel

0.20

In-2

Elliptio crassidens

35

1-20-67

9 months

Fish

< 0.14

In-2

Dorosoma cepedianuin

36

1-20-67

9 months

Mud

0.34

In-2

37

1-20-67

9 months

Mud

0.30

Out-2

38

1-20-67

9 months

Mud

0.49

In-3

42

1-20-67

9 months

Mud

0.30

"Control"

The C. W. England Laboratories converted 2,4-D and its esters to the methyl ester for reporting to TVA; however, for comparison with pub-
lished data on toxicity (2). all data were converted to the BEE equivalent.

2,4-D — 2,4-dichlorophenoxyacetic acid

, buloxyethanol ester
_, methyl esler

2 In=Inside embayment

Out=Outside in river channel external to embayment
Note: The station labelled "Control" received an unplanned application of

:,4-D and, therefore, cannot be considered a control.

Vol. 1, No. 3, December, 1967

19

Table 2. — Amilvscs of water sumples
WATTS BAR RESERVOIR
Goriton Braiuh Emhoxmcnt

^
c

\l KAIINITY

4-n

-

. I

ffi

z
o

e

I

uu

a.

5^5

<§

M^

75

u

<!)

Q

f J

H

no

c

a. -

H-

ta ^

<-

(Surface samples collecled

prior lo

2,4-D application)

A

3-17-«6 0805 55.7

10.0

8.1

0.0

70.1

B

3-17-«6 0825 56.J

10.2

8.6

1.4

35.9

—

—

C

3-17-66 0835 56.7

11.4

7.8 1

0.0

33.4

—

—

D

3-17-66 0840 56.2

10.9

8.1

0.0

34.9

—

E

3-17-66 0855 56.4

11.4

8.2

0.0

34.5

—

—

(Surface

samples collecled i hour after 2,4-D application)

A

3-18-66

0920

55.4

11.7

8.8

6.0

94.0

< 1

<1.45

B

3-19-66

0945

56.1

10.4

7.8

0.0

35.0

< 1

<1.45

C

3-19-66

0955

57.2

11.4

8.3

0.0

34.0

< 1

<1.45

U

3-19-66

1000

53.0

8.8

6.7

0.0

19.5

37

<1.45

E

3-19-66

1020

56.1

11.4

8.4

1.0

32.5

6

<1.45

(Surface samples collected 4 hours after 2,4-D application)

! 3-18-66

1220

59.0

12.8

9.1

18.0

92.0

<1

<1.45

3-19-66

12.10

61.6

10.8

7.9

0.0

35.0

6

<1.45

3-19-66

1240

59.3

11.5

8.5

2.0

36.0

< 1

<l.45

1

3-19-66

1245

62.4

12.6

8.6

3.0

36.0

< 1

<1.45

J

3-19-66

1305

56.8

10.4

8.5

1.5

35.0

2

<1.45

(Surface samples collcc

ed 8 hours after 2,4-D application)

A

3-18-66

1550

64.4

11.6

8.7

5.0

91.0

< I

<1.45

B

3-19-66

1510

64.7

10.4

8.2

0.0

36.0

< I

<1.45

C

3-19-66

1515

62.2

11.4

8.5

2.0

38.0

< 1

<1.45

D

3-19-66

1520

67.2

10.4

8.6

3.0

35.0

< 1

<1.45

E

(Sample missing)

TABLE 3. — Analysis af variance teslinr; differences in llie

mean numbers of Hexagenia between pre- and

pnyi-iicdlment of Watts Bar Reservoir with 2,4-D

Source of Variance

SS

Df

MS

F

Between four time periods

1,421.27

3

473.75

2.15

Between stations

6.155.24

20

307.76

1.38

Error

13,210.05

60

220.17

Total

20,786.98

83

N"": F 3,60,0.95 = 2 '^
20,60,0.95

All mud samples taken on the overbank were positive
for 2,4-D, with values ranging from 0.34 to 33.6 mg/kg
BEE. Only a single mud sample from the stations in the
channel had a concentration of BEE (0.30 mg/kg)
greater than that in the prestudy control (0.14 mg/kg
BEE) (Table 1 ).

Water samples from Ciunlersville showed a maximum
of 157 /,g/ 1 BEE I hour after application (Table 4).

BENTlllC FAUNA

Benthic fauna was sampled at 21 randomly located
stations within the 5-acre test area on Watts Bar Reser-
voir. Principal components of the Watts Bar benthic
fauna were mayflies (Hexagenia). midges (Tendipedidae),
biting midges (Heleidae), phantom midges (Chaohorus),
worms (Olii;ochacta). and Asiatic clams (Corhicula).

Submerged aquatic plants made excellent habitats for
aquatic insects. Elimination of watermilfoil would be
expected to affect the fauna associated with these plants.
Vannote (Unpublished TVA report. 1964. fn.wci survival
in reservoir areas treated with 2.4-D herbicide) showed
that two applications of 2,4-D in Guntersville Reservoir
did not depress benthic population densities, but the
eradication of watermilfoil eliminated a broad expanse
of substrate suitable for colonization by large popula-
tions of epiphytic insects such as the immature stages of
midges, mayflies, and dragonflies. This was also shown
in Watts Bar Reservoir where pre-treatment bottom
samples contained Anisoptera, Elmidae, Leptoceridae,
and Caenis. all of which were absent from 12-month
post-treatment samples.

Burrowing mayflies (Hexauenia) normally inhabit over-
bank substrate in TVA reservoirs and are a principal
component of the benthic fauna. Their abundance was
used as an index of the toxic effects of 2,4-D on benthos.
He.xagenia and other bottom fauna were sampled in
Watts Bar at 21 stations on 4 occasions. Analysis of
variance (Table 3) showed no significant change in the
abundance of Hexai;cnia in Watts Bar Reservoir between
one pre-treatment and three post-treatment sampling
periods (post 1 month. 10 months, and 12 months).
Density of Hexagenia populations was used to evaluate
effects of 2.4-D treatment on the benthic community
from the six stations sampled in Guntersville Reservoir.
Analysis of variance (Table 5) showed no significant
change in the mean numbers of Hexagenia between one
pre-treatment period and two post-treatment periods
(f)ost 1 month and 12 months).

SUMMARY

Results of this study niay be summarized as follows:

1. Application of 2,4-D herbicide in TVA's watermil-
foil control program caused no measurable toxic
elTect on benthic fauna in Watts Bar and Gunters-
ville Reservoirs.

2. Analysis of variance showed no significant change in
the mean numbers of burrowing mayllics (Hexagenia)
between pre- and post-treatment periods in Watts
Bar and Guntersville Reservoirs.

3. Bottom fauna samples indicated watermilfoil consti-
tuted the principal substrate inhabited by some in-
vertebrates. Elimination of substrate provided by the
watermilfoil resulted in loss of these invertebrates.

20

PusriClDES MONIIORING JOI'RN.M

TABLE 4. — Analyses of water samples

GUNTERSVILLE RESERVOIR

Vicinity of Comer Bridge

A
B
C
D

E
F

Alkalinity

Phenol. ' Total

(MG/1) (MG/1)

2,4-D

BEE

(;ig/I)

Acid

(„g/l as BEE)

(Surface samples collected prior to 2,4-D application)

A
B
C
D

3-29-66

1120
1110
1101

57.0
57.0
57.0
60.0

9.2
8.8

7.7
7.5

0.0
0.0

37.0
39.4

3
<0.5
7
7
6

<1.45
<1.45
<1.45
<1.45
<1.45

E

—

7

<1.45

F

1215

59.0

(Surface samples collected 1 hour after 2,4-D application)

4-5-66

0835
0845
0855
0925
0910
0905

56.5
56.0
56.3
56.0
56.0
56.0

_L

8.7
8.6
9.0
8.6
8.6

7.2
7.6
7.4
7.6
7.6
7.7

0.0
0.0
0.0
0.0
0.0
0.0

36.0
41.0
41.0
36.0
38.0
39.0

(Surface samples collected 4 hours after 2.4-D application)

(Surface samples collected 8 hours after 2,4-D application)

4-5-66

1455
1505
1515
1535
1530
1525

56.5
57.0
57.0
58.0
58.0
58.0

8.6
8.5
8.5
9.7
10.1
9.9

7.4
7.5
7.5
7.8
8.0
7.9

0.0
0.0
0.0
0.0
0.0
0.0

TABLE 5.— Analysis of variance testing differences in the
mean numbers of Hexagenia between pre- and post-
treatment of Gunlersville Reservoir with 2,4-D

SoiiRCE OF Variance

SS

Df

MS

F

Between three time periods

Between stations

Error

Total

62 M

775.33

519.27

1,357.04

2
5
10
17

31.22

155.07

51.93

0.60

2.97

Note:

F 2,10,0.95= l-\
^ 5,10.0.95 ~ '■''

91
157

36
5
5
3

_

—

—

—

—

—

—

—

—

9.2

7.6

0.0

36.0

8

9.3

7.6

0.0

37.0

130

9.4

7.8

0.0

36.0

19

<1.45
<1.45
<1.45
<1.45

<1.45
<1.45

<1.45
<1.45
<1.45

4. There was little uptake of 2,4-D by fish but some
by mussels.

5. Significant concentrations of 2,4-D were noted in
iso!ate(i sediment samples up to 10 months after
treatment.

Conclusion

These data indicate that high application rates of 2,4-D
for watermilfoil control on TVA reservoirs have not

Vol. 1, No. 3, December. 1967

produced adverse effects on aquatic fauna or water
quality.

LITERATURE CITED

(/J Snuth, Gordon £., T. F. Hall. Jr., and R. A. Stanley.

1967. Eurasian watermilfoil in the Tennessee Valley.

Weeds 15: (2).
(2) Mount, D. I. and C. E. Stcphan. 1967. A method for

establishing acceptable toxicant limits for fish — mala-

thion and the butoxyethanol ester of 2,4-D. Trans.

Amer. Fish Soc. 96(2) : 185-193.

21

PESTICIDES IN SOIL

Monitoring for Chlorinated Hydrocarbon Pesticides in Soil
and Root Crops in the Eastern States in 1965

W. L. Seal', L. H. Dawsey=, and G. E. Cavin'

ABSTRACT

Forty-nine fields plumed to root crops were sampled in the
fall of 1965 to determine levels of chlorinated hydrocarbon
pesticide residues in soils and crops. Selection of fields was
based on the relatively heavy use of persistent insecticides
in prior years. Materials analyzed in the study weie soil,
potatoes, carrots, and peanut meats. The methods of analy-
sis employed were gas chromatography (electron capture)
and thin layer chromatography.

DDT was found in soil in 48 of the 49 fields sampled,
ranging from 0.10 to 12.8 ppm, and averaging 2.8 ppm.
Residues of DDT were well below the tolerance levels in
all crop samples. Dieldrin was present in soil in 28 of the
49 fields sampled, ranging from 0.05 to 0.26 ppm. No diel-
drin residues were delected in potato tubers, and residues
of the chemical averaged 0.05 ppm in 6 of 19 composite
carrot samples. Dieldrin was present in all five composite
peanut meat samples, averaging 0.10 ppm. Sampling was
too limited, however, to draw any conclusions as to whether
this contamination of peanuts is a significant problem. Ad-
ditional monitoring was conducted in 1966 and 1967.

Introduction

A limited study was conducted in the fall of 1965 in
seven Eastern States to determine levels of chlorinated
hydrocarbon pesticide residues in food and soil from
land treated with persistent pesticides. Potatoes, carrots,
and peanuts were selected for this study since these root
crops are more readily contaminated by pesticide resi-
dues in the soil through adsorption or translocation.

A preliminary survey of appropriate fields was conducted
prior to sampling. Selection of fields to be sampled was
based on the relatively heavy use of persistent insecti-
cides since 1961, particularly chlorinated hydrocarbons,
for the control of insect pests. All of the fields selected
were treated with persistent pesticides at least 1 or more

1 Plant PcM Comrol Division, Agricultural Research Service, Hyatts-

ville. Md 20782
- Plant Pest Control Division, Agricultural Research Service, Gulfport,

MiM. 39501.
3 Plant Pest Comrol Division, Agricultural Research Service, Moores-
town. N. J. 08057.

years during this period. Of 49 fields selected, 25 were
planted to potatoes, 19 to carrots, and 5 to peanuts.

Sampling Methods
Three 1-acre plots were laid out in each field on a strati-
fied random basis. The plots were located in relation to
drainage and other topographical features. Fifty soil
cores, 2 inches in diameter and 3 inches deep, were taken
from within the rows in each l-acre plot. Potato, carrot,
and peanut samples were collected at the same time and
place the soil cores were taken.

The soil cores from each plot were placed in a large
container and passed through a l^-inch screen to facili-
tate mixing. Stones, roots, and other trash that would
not pass through the screen were discarded. A new
1 -gallon paint can was then filled with the mixed, screened
soil and sealed with an airtight lid. Each container was
labeled with a field sample number and date. Extreme
care was taken to thoroughly clean the sampling equip-
ment after each sample was collected.

The containers of soil and bagged crop samples were
stored at room temperature until they could be shipped
to the laboratory at Gulfport, Miss.

Analytical Procedures
The sensitivity limits of the analytical procedures used
were generally 0.05 ppm for residues in soil and 0.01
ppm for residues in crops.

Three hundred grams of soil were tumbled for 4 hours
with 600 ml of a 3:1 mixture of hexane and isopropanol
of chromatographic grades. The isopropanol was re-
moved by repeated washing with distilled water and the
washed extract was dried by filtration through anhydrous
sodium sulfate. A representative aliquot was stored under
refrigeration in a sealed glass bottle prior to direct deter-
mination of pesticides in the extract by gas chromatog-
raphy. Another 100-g lot of soil was dried overnight in
an oven at 1 10 C for parallel determination of the mois-
ture contents of the soil at the time of analysis for pesti-
cides.

22

Pesticides Monitoring Journal

Preparation of samples and analytical procedures used
were the same for carrots and potatoes. Samples were
water-washed and air-dried before chopping. A vertical
segment of each root or tuber was included in the first
sample size reduction which was accomplished in a food
chopper. One hundred grams of the pulp was homoge-
nized for 1-2 minutes with 200 ml of chromatographic
grade acetonitrile in the 450-ml stainless steel cup of a
Lourdes Multimix Homogenizer (MM-l)^ with about
lOg of Celite added. The solvent and pulp were sep-
arated by centrifuging in the cup. The acetonitrile was
decanted through a filter paper into a 1 -liter flask and
held. Another 100-ml portion of acetonitrile was added
to the pulp in the cup: the extraction, settling, and filtra-
tion were repeated, the second extract being added to the
first in the flask. The total volume of acetonitrile was
then reduced by evaporation through a Snyder column
on a hot plate, to leave a water layer covermg the bot-
tom of the flask. One hundred milliliters of hexane was
added through the column to the water layer; the hexane
was evaporated completely, carrying with it remaining
traces of acetonitrile from the water. Another 200-ml
portion of hexane was added through the Snyder column
to the water layer in the flask, and the contents were
refluxed to insure complete solution of the pesticides in
hexane. Water and hexane were transferred to a separa-
tory funnel where the water was rejected. The hexane
extract Wis filtered through sodium sulfate, made to
300-ml volume, sealed in a glass bottle, and held under
refrigeration until final determination was made by gas
chromatography. Cleanup was unnecessary with carrots
and potatoes when the initial extraction was made with
acetonitrile.

The harvested peanut pods had been air-dried according
to farm practice. Shells were removed by hand at the
laboratory and discarded. Pesticides which might have
passed from the shells to meats in handling were elim-
inated by rinsing the meats, first with an isopropanol
wash, then with a hexane wash, prior to grinding of
samples. The washed meats were ground dry in a blender
to give a free-flowing meal, from which a 20-g aliquot
was weighed. The meal was homogenized with 100 ml
of isopropanol in the Lourdes Multimixer (see above).
The mixture was washed into a Mason jar with 300 ml
of pentane, timibled for 2 hours, then allowed to settle.
The extract was decanted through a filter into a separa-
tory funnel, and the isopropanol was removed by re-
peated water washes. The peanut oil was eliminated from
the pentane extract by means of the acetonitrile partition
method. This method consisted of reducing the volume
to 50 ml by evaporation of pentane, transferring the
extract to a 125-ml separatory funnel, adding an equal
volume of acetonitrile (saturated with pentane), equili-

■• Lourdes Instrument Corp., 656 Montank Ave., Brooklyn, N. Y. 11208.

Vol. 1, No. 3, December, 1967

brating, settling, and drawing off the acetonitrile into a
250-ml separatory funnel. The pentane in the first
separatory was washed two more times with acetonitrile
to extract all pesticides from the pentane which was
rejected. The combined washings in the second separa-
tory were backwashed with 40 ml of hexane which was
rejected, and the acetonitrile was transferred to a 500-ml
f-jointed flask for evaporation and transfer of pesticides
back into fresh hexane. Transfer back to hexane was
accomplished using the same procedures as for the carrot
extracts.

Up)on completion of partitioning for removal of fat, the
extract was divided in half, which was equivalent to
lOg of the original peanut meats, and the half-extract
was concentrated by evaporation to 5.0-ml volume pre-
paratory to cleanup by column chromatography. The
glass chromatographic column, 450 mm x 10 mm ID,
resembled an ordinary 50-ml burette with teflon stop-
cock at the bottom. Prior to operation, the column was
filled about halfway with 10 g of Florex absorbent
(AARVM 60/100 mesh, activated at 130 C for 16
hours); the absorbent was pre-washed first with 50 ml
of 10% ether in hexane, then with 50 ml of hexane.
The 5.0 ml of concentrated extract was then passed
through the absorbent elution with 150 ml of 10% ether
in hexane, which solvent was caught in a 250-ml Ku-
derna-Danish evaporator at the bottom of the column.
The evaporator was fitted with a 15-ml graduated
centrifuge tube, which enabled the eluate to be re-
concentrated to a 5.0-ml volume after placing the evap-
orator on a steam bath. This final cleanup sample was
sealed by means of a glass stopper in the centrifuge
tube, pending determination of pesticides by gas chroma-
tography.

Unknown residues in the above described hexane ex-
tracts were determined by injecting 2.5- to lO.O-juliter
portions into columns of gas chromatographs followed
by interpretation of the tracings made on the record
charts, as compared with similar tracings made from
injection of known amounts of pesticides. Columns used
were as follows:

DC-200, 3%, on Gas-Chrom-Q (100-120 mesh)

QF-1, 5%, on Diatoport-S (100-120 mesh)

SE-30, 5%, on Chromosorb-W ( 60-80 mesh)

Dow-11, 5%, on Chromosorb-W ( 60-80 mesh)

Chromatographic instruments employed were the Jarrell-
Ash 28-730 and the F & M 810, each equipped with
two columns and two electron capture detectors. Typical
operating conditions for the DC-200 column as in-
stalled in the F & M 810 instrument were as follows:

Column : All glass, 8 feet x 3 mm ID

Gas: Methane- Argon, at 120 ml/ min for

inlet pressure of 60 lb

23

Sensitivity: 5.12x10"
Temperatures: Column 180 C

Detector 2 1 0 C

Sampler 235 C
Chart speed: 15 inches/ hour

The other three columns may have been operated under
conditions somewhat different from this particular one;
however, portions of extract were injected in at least
two different columns, sometimes four different columns,
to verify the identities of pesticide peaks produced on
the charts. When identity of a peak appeared doubtful
on two or more charts, further confirmation was ob-
tained by thin layer chromatography methods.

Soil, potatoes, carrots, and peanut meals were analyzed
in different groups. Each group was organized with a set
of five controls before starting the materia! through the
various analytical steps. The five controls built into each
group of samples at the start were as follows: (1) A
9-component pesticide standard, diluted with the extrac-
tion solvent, was prepared and bottled for calibrating use
in the final gas chromatographic determinations; (2)
A portion of the extraction solvent only was carried
through all analytical steps to detect pickup of extraneous
substance, if any: (3) An extraction solvent fortified
with pesticides, same as the first calibrating standard, was
carried through the analysis; (4) A composite sample
was prepared from portions of each sample in the group
to be analyzed; and (5) A composite sample was pre-
pared similar to the fourth control but fortified with the
same pesticides as were added to the calibrating standard
(first control) and the fortified solvent (third control).
The last four controls were carried through all analytical
steps for the purpose of determining overall recovery of
pesticides, both with solvent alone and with the actual
material under analysis.

Recoveries of dieldrin. endrin, heptachlor, and members
of the DDT-complex from soils, ranged from 87% to
107% with composite control samples. Soil residues were
corrected with appropriate recovery factors applying on
individual groups of samples, and also for moisture con-
tent of each sample. With potatoes, recovery of the
DDT-complex was 134%, dieldrin — 120%, endrin —
100%, and heptachlor — 97%. With carrots, recovery of
the DDT-complex was 87%, dieldrin — -85%, endrin —
77%, and heptachlor — 73%. Efficiency of analysis of
the peanut meats was less than that of soil, potato, and
carrot samples, inasmuch as cleanup involving partition-
ing and Florex column treatment was employed. Recov-
eries from composite samples of peanuts were 50% for
the DDT-complex. and 47% for dieldrin. No controls
with endosulfan were incorporated in the groups, and the
endosulfan found present in certain of the soils, but not
the crops, was based on a bench standard. Such residues
were reported without correction for losses.

Results ami Discussion

DDT was found in the soil in 48 of the 49 fields, averag-
ing 2.8 ppm. Few analyses were above 6 ppm DDT.
The highest found were 12.8 and 9.5 ppm in samples
from two carrot fields and 7 ppm in one potato field.
DDT residues in potatoes and carrots were well below
tolerance levels. All carrot samples contained some
DDT, however, and DDT residues were found in po-
tato samples from 21 of 25 fields but in extremely low
amounts. Residues of DDT in soil in peanut fields aver-
aged 0.3 ppm, and residues in peanut meats averaged
0.05 ppm in samples that contained detectable residues.

Dieldrin was present in soil samples in all 5 peanut fields
sampled; 19 of 25 potato fields; and 4 of 19 carrot fields.
No measurable residues were found in potatoes, and
residues in carrots did not exceed 0.14 ppm. Residues in
peanut meats averaged 0.10 ppm.

Treatment histories indicate that aldrin, which converts
to dieldrin, was used in prior years on all five of the
peanut fields. The most recent treatment of record was
on one field where 40 lb of 5% dust or 2 lb of technical
aldrin were applied per acre. Residues in peanuts from
this field were 0.13 ppm. In another field, some dieldrin
residues were found in peanuts even though the last
known application of aldrin was in 1953, at the rate
of 1.5 lb actual per acre. A study of the limited residue
data showed that dieldrin residues in peanut meats were
roughly equal to about two-thirds of the residues in the
top 3 inches of soil where the crop was grown.

Endrin was found in the soil in about one-fifth of the
fields. Residues were not found in crop samples using
a method sensitive to 0.01 ppm.

Heptachlor and/or heptachlor epoxide was found in the
soil in four potato fields and five carrot fields. No resi-
dues were detected in potatoes, but residues averaged
0.07 ppm in two carrot samples.

Endosulfan has been applied to potatoes in the past few
years. It is replacing some of the chlorinated hydrocar-
bons previously used in the control of certain pests. En-
dosulfan residues averaging 0.46 ppm were found in
soil from 23 of the 25 potato fields. No residues were
detected in tubers, however. Endosulfan was not used
on the peanut fields sampled and on only 1 of the 19
carrot fields.

Summary

DDT ranged from 0.10 to 12.8 ppm in 48 of the 49
fields that were sampled in 1965. Based on the data
developed in these studies, it appears that DDT residues
in soil in these fields should not result in residues above
presently accepted levels for potatoes, carrots, and
peanuts.

24

Pesticides MoNiroRiNC Journal

TABLE 1. — Pesticide residues in soil and root crops collected in the Eastern States in 1965

No. OF
Fields

Material
Sampled

DDT

Dieldrin

Endrin

Heptachlor and/or
Heptachlor epoxide

Endosulfan

Crop

Positive
Com-
posite
Samples^

Residue

(ppm)

Range &

Average

Positive
Com-
posite
Samples^

Residue

(ppm)
Range &
Average

Positive
Com-
posite

Samples'

Residue

(ppm)

Range &

Average

Positive
Com-
posite
Samples'

Residue

(ppm)

Range &

Average

Positive
Com-
posite
Samples'

Residue

(ppm)

Range &

Average

Peanuts

5

Sou

5

0.10-0.71
(0.30)

5

0.08-0.20
(0.15)

0

0

0

Meats

3

0.02-0.07
(0.05)

5

0.08-0.13
(0.10)

Potatoes

25

Soil

24

0.33-7.04
(2.75)

19

0.08-0.20
(0.10)

8

0.08-0.50
(0.27)

4

0.05-0.10
(0.08)

23

0.08-1.17
(0.46)

Tubers

21

0.01-0.06
(0.02)

0

0

0

0

Carrots

19

Soil

19

0.49-12.8
(3.67)

4

0.05-0.26
(0.19)

2

0.05-0.10
(0.08)

5

0.06-0.26
(0.16)

1

0.49

Roots

19

0.20-2.31
(0.85)

6

0.01-0.14
(0.05)

0

2

0.06-O.08
(0.07)

0

1 Represents results from analysis of one composite sample per field.

DDT 1 , 1 , l-trichloro-2,2-bis ( p-chlorophenyl ) ethane

Dieldrin 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-endo-p.To-5.8-dimethanonaphthalene

Endrin l,2,3,4,I0,10-hexachloro-6,7-epoxy-l,4,4a,5.6,7,8.8a-octahydro-l,4-e'«rfo-enrfo-5,8-dimethanonaphthalene

Heptachlor 1,4,5,6,7, 8,8-heptachloro-3a, 4, 7. 7a-tetrahydro-4,7-methanoindene

Heptachlor epoxide 1 ,4,5,6,7,8,8-heptachloro-2,3-epoxy-3a,4,7,7a-tetrahydro-4,7-methanoindan

Endosulfan 6,7,8,9.10,10-hexachloro-l,5,5a,6.9.'>a-hexahydro-6,9-methano-2,4,3-benzodioxathiepin 3-oxide

Dieldrin residues were not found in potatoes grown on
soil previously treated with aldrin or dieldrin, but pre-
liminary investigations indicate low-level dieldrin resi-
dues (0.10 ppm) in peanut meats. The limited sampling
that has been done is not sufficient to serve as a basis
for any definite conclusion as to whether this is a sig-

nificant problem. Further testing was conducted in 1966
and 1967 to confirm these results.

Trade names and the names of commercial companies are used in this
paper solely for the purpose of providing specific information. Mention
of a trade product or manufacturer does not constitute a guarantee or
warranty by the U. S. Department of Agriculture or an endorsement by
the Department over other products or manufacturers not mentioned.

Vol. 1, No. 3, December, 1967

25

Infonnaiion for Contributors

The Pesticides Monitoring Journai welcomes from
all sources qualified data and interpretive information
which contrihute 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
sampling and anaUtical 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-
laboratory checks. The procedure employed should be
referenced 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 Styi E Manual for Biological
Journals. American Institute of Biological
Sciences, Washington. D. C, and/or the Style
Manual of the United States Government Print-
ing Ofl^ice.

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 fiat form — not folded or rolled.

Manuscripts should be typed on 8', 2 x II inch

paper with generous margins on all sides, and
each page should end with a completed para-
graph.

. All copy, including tables and references, should

be double spaced, and all pages should be num-

bered. The first page of the manuscript must
contain authors" full names listed under the title,
with afflliations, 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 alpha-
betically, giving name of author/s/, year, full title
of article, exact name of periodical, volume, and
inclusive pages.

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 he 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
1 00 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, notation of such
should be provided.

Correspondence on editorial and circulation matters
should be addressed to: Mrs. Sylvia P. O'Reur. Editorial
Manager, Pesticides Monitoring Journal. Pesticides
Program, National Communicable Disease Center, At-
lanta, Georgia .^O.V^.V

26

Pesticides Monitoring Journal

The Pesticides Monitoring Journal is published quarterly under the auspices of the Federal
Committee on Pest Control and its Subcommittee on Pesticide Monitoring as a source of
information on pesticide levels relative to man and his environment.

The parent committee is composed of representatives of the U. S. Departments of Agriculture,
Defense, the Interior, and Health, Education, and Welfare.

The Pesticide Montoring Subcommittee consists of representatives of the Agricultural Research
Service, Consumer and Marketing Service, Federal Extension Service, Forest Service, Depart-
ment of Defense, Fish and Wildlife Service, Geological Survey, Federal Water Pollution
Control Administration, Food and Drug Administration, Public Health Service, and the
Tennessee Valley Authority.

Responsibility for publishing the Pesticides Monitoring Journal has been accepted by the
Pesticides Program of the Public Health Service.

Pesticide monitoring activities of the Federal Government, particularly in those agencies repre-
sented on the Pesticide Monitoring Subcommittee which participate in operation of the national
pesticides monitoring network, are expected to be 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 nongovernment research
institutions, both within and without the United States. 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 Subcommittee. Authors are given the benefit of review comments prior to
publication.

Editorial Advisory Board members are:

Reo E. Duggan, Food and Drug Administration, Chairman

Andrew W. Breidenbach, Public Health Service

Anne R. Yobs, Public Health Service

James B. DeWitt, Fish and Wildlife Service

S. Kenneth Love, Geological Survey

Milton S. Schechter, Agricultural Research Service

Paul F. Sand, Agricultural Research Service

Trade names appearing in the Pesticides Monitoring Journal are for identification only and do
not represent endorsement by any Federal agency.

Address correspondence to:

Mrs. Sylvia P. O'Rear

Editorial Manager
PESTICIDES MONITORING JOURNAL

Pesticides Program

National Communicable Disease Center

Atlanta, Georgia 30333

CONTENTS

Volume 1 March 1968 Number 4

Page
EDITORIAL 1

RESIDUES IN FOOD AND FEED

Pesticide residues in vcfictable oil seeds, oils, and by-

products 2

R. E. Duggan

Investifintion of lead residues on growing fruits and

\egetables 8

Abram Kleinman

Pesticide residues in total diet samples (III) 11

R. J. Martin and R. P.. Duggan

RESIDUES IN FISH, WILDLIFE, AND ESTUARIES

Pesticide monitoring of the aquatic biota at the Tule

Lake National Wildlife Refuge 21

Patrick J. Goclsil and William C. Johnson

Chlorinated pesticide residues in an aquatic environ-
ment located adjacent to a commercial orchard 27

R. J. Moiibry. J. M. Helm, and G. R. Myrdal

PESTICIDES IN SOIL

Monitoring the effects of the I9f>3-fi4 Japanese Beetle
control program on soil, water, inul silt in the Battle

Creek area of Michigan 30

J. E. Fahey, J. W. Butcher, and M. E. Turner

EDITORIAL

The increasing number of pesticide monitoring programs
magnifies tiie difficulty in evaluating the results of in-
dividual studies and of comparing them with earlier
studies.

The list of factors contributing to the problem is long
and includes such items as differences in experimental
design; lack of adequate experimental controls; insuffi-
cient knowledge concerning the chemical characteristics
of pesticides; differences in sample collection, handling,
and storage; variations in efficiency of cleanup pro-
cedures; differences in sensitivity of chemical analytical
procedures; use of chemicals of inadequate purity as
controls; technician variation; and so on.

Each could quite properly serve as the subject of an
editorial. However, the present discussion is limited to
the uncertainty introduced into pesticide monitoring
data by the use of analytical methods of unknown
reliability and the difficulty in comparing resuhs on
similar systems when different cleanup and analytical
methods are used. Certainly no one is against progress,
and changes in methodology to improve sensitivity,
resolution, or recovery are necessary. However, constant
alteration of methodology must lead to confusion.
Ideally, a sensitive, reliable, and reproducible analytical
procedure should be adopted for each substrate studied.
The analytical procedure should be standardized and
fully evaluated in order to serve as a reference in eval-
uating future modifications or entirely new procedures.
This is equally important for sampling techniques, clean-
up procedures, and instrumental analysis, including
interpretation of tracing. Procedures used for closely
related substrates should then be compared.

There are those who would argue that such standardiza-
tion is not necessary because the same technique does
not work equally well for every laboratory; and that,
therefore, each laboratory should use its best technique.
This, of course, is just what has been happening; hence
the current difficulties. Such an approach is characterized

by the statement, "We use the procedure

but with certain modifications." Sometimes it seems
everyone has his own set of modifications!

Development of the gas chromatograph and of the elec-
tron capture and other detectors has been a boon to
pesticide residue chemistry. This chemical specialty has
long been an art; it is time to add standardization and
make it a science.

Anne R. Yobs

Member, Editorial Advisory Board

Vol. 1, No. 4, March 1968

RESIDUES IN FOOD AND FEED

Pesticide Residues in Vegetable Oil Seeds,
Oils, and By-Products

R. E. Duggan'

Earlier reports (1-3) have discussed, in terms of broad
food categories, the pesticide residue data obtained by
the Food and Drug Administration in surveillance and
monitoring programs conducted from July 1, 1963
through June 30, 1966. The findings on fluid milk and
other dairy products have been reported in considerable
detail (4).

The principal purpose of this paper is to report and
evaluate the findings on samples of products derived
from oil seed crops. Since these crops constitute an
important segment of the Nation's food supply, pesticide
residues incurred in their production are of substantial
importance. Direct additions of these pesticide residues
to man's diet may occur from consumption of these
crops which have been treated with pesticides in their
production. Indirect additions to man's diet from these
crops may occur from the use of by-products in the
production of milk, meat, and poultry; and some tol-
erances have been established on this basis. Additionally,
man may receive residues by consuming foods con-
taminated through drift and runoff and through crop
rotation — for example, the planting of soybeans in
areas previously treated for cotton production. It may
be impossible currently to measure the relative effects
of the various factors influencing the incidence and
levels of pesticide residues in food. However, there is a
need to establish, with a reasonable degree of certainty,
the major factors making up the total residue content
of the food chain.

Portions of several major program divisions, such as
raw agricultural products, processed animal feeds,
vegetable oils, and processed foods, have been excerpted
for this report.

' OITicc of Associate Commissioner for Compliance. Food. :md Driin
Adminislralion. U. .S. Dcp.irlmcnl of Health. Education, and Welfare,
Washington, D. C. 20204.

Sampling Procedures

Samples were collected on a nationwide basis as a part
of the Food and Drug Administration's surveillance
program carried out in 18 District offices. Samples col-
lected in surveillance programs are classified as "ob-
jective" if "unknown" with respect to the possibility
of excessive residue content or actual misuse of pesti-
cide chemicals. The selection of sampling points and
scheduling of samples was left to the discretion of the
I 8 District offices.

Specific lots were sampled (5) for analysis by taking
several portions from the lot. The portions were com-
bined for analysis.

Laboratory A nalysis

Generally, samples were examined promptly after collec-
tion. All analyses were made in FDA District Lab-
oratories by multiple residue gas-liquid chromatographic
methods. All of the laboratories concerned participated
in method validation studies reported by Johnson (5),
Krau.se (9), Gaul [10), and Wells (//). Electron
capture and microcoulometric detectors were employed.
The methods used during this investigation were basi-
cally those which have become official A.O.A.C. pro-
cedures (6); the detailed procedures employed are
described in the FDA Pesticide Analytical Manual, Vol.
I (7). Quantitative sensitivity limits for gas chromato-
graphic analysis, readily attainable on most products in
normal laboratory operations, were based on Vi full-scale
detlections (1 X 10"° Amperes Full-Scale) for 1 ng of
aldrin and, for program purposes, were established at
0.05 ppm for raw agriculture products, and at 0.25 ppm
(fat basis) for fatty foods. Confirmatory analyses by
thin layer chromatography were made when results
exceeded these figures. Quantitative results below these

Pesticides Monitoring Journal

levels were reported but were not confirmed by check
analysis, and are recognized as having reduced accuracy
limitations common to all quantitative estimations at the
lower ranges of method sensitivity.
Recoveries, in general, range between 80'^ and 110%
for most pesticide residues and commodities. No correc-
tions for recovery have been made, and the values are
reported on an "as is" basis.

Results

The data are not amenable to evaluation on a geographic
or production basis. Inspection of the raw data indicates
that samples were reasonably well distributed according
to major program divisions among the 18 District
offices.

A total of 1,230 residues of 20 pesticide chemicals were
reported in 641 positive samples of the 2,389 samples of
raw products, meal, crude oil, refined oil, and oleo-
margarine. DDT and its analogues (DDE and TDE),
dieldrin, lindane, toxaphene, endrin, BHC, and chlordane
account for 95% of the residues found in oil seeds,
96% of the residues in oil seed meals, 98% of the
residues in crude oils, and 95% of the residues in re-
fined oils. Malathion residues were found only in the
raw cottonseed, peanuts, and soybeans. Aldrin and
heptachlor epoxide were found too infrequently to be
considered significant. Seven other pesticide chemicals
were found in one or two samples.

Table 1 shows the percent of residues at arbitrarily
selected ranges in the raw products, meals, crude oil,
refined oil, and oleomargarine based on the total number
of instances in which residues were found. Most of the
residues were found at low levels: 94.5% of the values
were below 0.51 ppm, and 75.5% of the values were
below 0.11 ppm. This general pattern is observed re-
gardless of the commodity, product, or individual pesti-
cide chemical involved.

Tables 2-5 summarize the incidence and average levels
of specific pesticide chemicals found in soybeans,
peanuts, cottonseed, corn, and products derived from
the raw commodity. Although corn is not generally
classified as an oil seed, all available data have been
included since the production of corn oil is substantial.
The samples represent grain corn generally and are not
confined to that used for the production of corn oil.

During the period covered in this report, 53 objective
samples of oleomargarine were collected as shown in
Table 6. Of these samples. 18.9% contained DDT,
5.1% contained TDE, 7.5% contained DDE. and two
samples (3.8%) contained BHC.

The number of objective samples examined in some
product classes, such as refined peanut and cottonseed
oil, are too few to be considered representative of the
surveillance period involved and therefore have limited
usefulness.

The average level for each pesticide residue in Tables
2-6 includes all samples and was calculated by using the
midpoint of each range and the percent of samples
falling in the range. The actual values were used for
those residues exceeding 2 ppm.

No significant trends were observed on an annual basis
where the number of samples was large enough for
consideration of trends.

Discussion

Legal tolerances have been established for some of the
chemicals found in the raw agricultural product as
follows :

Residues in PPM

Cotton-
seed

Soybeans

Peanuts

Grain
Corn

DDT

Dieldrin
Toxaphene
Endrin
Chlordane

4

5
zero

1.5
zero

2

7
7
3

zero

Except for the positive findings of dieldrin in soybeans,
there were no samples containing pesticide chemicals
exceeding the tolerance level. Over 60% of the residues
commonly present are not sanctioned by tolerances in
the raw agricultural product.

The kinds, incidence, and levels of pesticide residues in
soybeans, cottonseed, peanuts, and corn are quite similar.
Although the tabular data indicate higher average levels
of all residues except endrin in cottonseed, the small
number of cottonseed samples involved does not permit
a high degree of reliability in this observation.
It must he recognized that peanuts and corn may be
eaten unprocessed and that processing procedures would
significantly change the pesticide residue content of the
product. Cottonseed and soybeans, on the other hand,
generally undergo processing into other products before
consumption.

Table 7 compares the pesticide chemicals found in the
various oil seeds and oil seed products, including oleo-
margarine. Data from the "Oils, Fats, and Shortening"
portion of the Total Diet studies also are included in
this table for comparison.

Average residue values are higher in crude oils than in
the raw products or meals. The average levels of pesti-
cide residues in soybeans, cottonseed, and peanuts are
quite low when compared to the established tolerances;
for example, the average of 0.15 ppm DDT compared
to the 4 ppm tolerance on cottonseed, or 0.03 ppm DDT
compared to the 7 ppm tolerance on peanuts.
The incidence of residues within various quantitative
ranges shows that 78% of the values on oil seed are
below 0.11 ppm, and 98% are below 0.51 ppm. The

Vol. 1, No. 4, March 1968

incidence of residues in griiin corn at these levels was
96'"c and lOO"";, respectively.

The residue content of oil seed meals is important be-
cause of the general use of such products in animal feed.
The average levels of pesticide residues in oil seed
meals or cakes are low. The distribution of residues
within various quantitative ranges shows that 87% of
all residues were below 0.1 I ppm, and 96.5% were
below 0.51 ppm.

As expected, the average values of residues in crude
oils are much higher than in the other products. Since
the results for oil seeds are reported on an "as is" basis,
the higher values for the oil content of the product
under examination must be considered. The incidence of
residues within various quantitative ranges shows that
61% of the values were below 0.1 1 ppm, and 28% were
between 0.11 ppm and 0.50 ppm.

After refining, the average levels of residues are sub-
stantially lowered and are similar to the average values
found in oleomargarine.

The incidence of residues in the various quantitative
ranges shows 56% of the values below 0.11 ppm, and
31% between 0.11 ppm and 0.50 ppm. However, no
values in excess of 1.50 ppm were reported in refined
oils compared to 1.8% of the values in crude oils.

Except for endrin, residues of the pesticide chemicals
most commonly found in the raw product were also
found in the refined oil at considerably lower levels.
Endrin was not found in refined oils. Only residues of
the DDT compounds and BHC were found in oleo-
margarine.

The average levels found in these samples of refined oils
are somewhat higher than those found in the 70 com-
posites of oils, fats, and shortening from the total diet
samples examined during the period June 1964 through
April 1967. Residues of toxaphene and chlordane were
not reported in the total diet composites. The oils, fats,
and shortening composite is prepared from salad dress-
ings, mayonnaise, salad oil, shortening, and peanut
butter.

Summary and Conclusions

Residues of DDT and its analogues (TDE and DDE),
dieldrin, lindane, toxaphene, endrin, BHC, and chlor-
dane were frequently found in vegetable oil seeds and
products.

Residues of other pesticide chemicals were not found
with sufficient frequency to be considered significant.

None of the residues found in oil seeds exceeded the
tolerances where finite tolerances have been established.
Endrin residues were found in cottonseed, for which the

established tolerance is zero. Dieldrin residues were
found in soybeans and grain corn which have an estab-
lished tolerance of zero. Over 60% of the residues found
were not sanctioned by tolerances in the raw agricultural
product.

Chlorinated organic pesticide residues are relatively
high in the crude oil. Significantly lower values were
found in the refined oils and in the oil seed meals and
cakes.

While the residue levels found in these samples indicate
that oil seeds and products do not present a serious
problem, it is obvious that, when such residues are
present — whether from approved applications, misuse,
or unavoidable sources — the finished product will prob-

TABLE 1. — Distrihiilion of residues, hy product, in difjereni
quantitative ranges

[T = <0.001 PPM]

Percent

OF Positive Samples

Level

Crude

Refined

Oleomar-

(PPM)

Seed

ME4L

Oil

Oil

garine

T-0.03

65.2

71.4

43.0

33.3

68.4

n.04-0.10

17.8

15.1

17.7

23.1

10.5

11.11-0.50

15.8

10.1

28.5

23.1

21.1

0.51-1.00

1.0

2,1

6.6

7.7

1.01-1.50

0.2

0.8

2.4

12.8

1.51-2.00

0.4

0.5

>2.00

1.3

TABLE 2. — Incidence of specific pesticide residues in
soyhciin products

[T = <0.001 PPM; — = Not detected]

Percent of Samples Containing Specific Pesticides
AND Average Level (PPM) of Each Pesticide

Soybeans

Crude
Oil

Meal
(Cake)

Refined
Oil

DDT

9.6

(0.006)

16.3
(0.015)

7.0
(0.005)

13.0
(0.003)

TDE

0.7
(T)

7.1
(0.006)

1.4
(T)

—

DDE

2.7
(T)

6.1
(0.002)

3.5
(0.001)

—

Dieldrin

8.0
(0.002)

17.3
(0.013)

1.4
(T)

4.3
(T)

Lindane

2.2
(T)

2.0

(T)

4.9
(0.001)

—

Toxaphene

8.0
(0.004)

4.1
(0.024)

—

4.3
(T)

Fndrin

9.8
^ 0.008)

6.1

(0.013)

0.7
(T)

—

ItHC

2.9
(T)

11.2
(0.004)

0.7
(T)

—

Chlordane

0.9

(T)

0.7
(T)

—

Total Number
of Samples

550

98

143

23

Percent

w, residues

26.7

34.7

14.0

17.4

Pesticides Monitoring Journal

ably contain a portion of the pesticide chemical. When
these residues are added to other unsanctioned additions
in the total diet, they may eventually reach a total level
that will have an impact on the existing tolerances for
residues on raw agricultural products generally.

TABLE 3. — Incidence of specific pesticide residues in
peanut products

[T = <0.001 PPM; ■

: Not detectedj

Percent of Samples Containing Specific Pesticides
AND Average Level (PPM) of Each Pesticide

Nuts

Crude
Oil

Meal
(Cake)

Refined
Oil

DDT

13.5
(0.025)

66.7
(0.466)

45.2
(0.140)

20
(0.060)

TDE

1.7

(T)

41.7
(0.128)

22.6
(0.026)

20
(0,007)

DDE

9.6
(0.001)

58.3
(0.909)

38.7
(0.025)

20

(0.032)

Dieldrin

8.5
(0.008)

22.2
(0.017)

22.6
(0.006)

20
(0.007)

Lindane

2.8
(0.002)

5.6

(0.001)

3.2
(T)

—

Toxaphene

1.7
(0.006)

2.8
(0.008)

—

—

Endrin

1.1
(T)

2.8
(0.008)

—

—

BHC

3.4
(0.007)

8.3
(0.002)

12.9
(0.006)

10
(0.0(J2)

Total Number
of Samples
Percent

w/Residues

177
26.6

36
75.0

31
61.3

10
33.3

TABLE 4. — Incidence of specific pesticide residues in
cottonseed products

[T = <0.001 PPM; — = Not detected]

Percent of Samples Containing Specific Pesticides
AND Average Level (PPM) of Each Pesticide

Seed

Crude
Oil

Meal
(Cake)

Refined
Oil

DDT

69.5
(0.154)

29.2
(0.077)

28.5
(0.028)

12.2
(0.024)

TDE

13.0
(0.029)

35.4
(0.093)

12.9
(0.003)

17.1
(0.016)

DDE

13.0
(0.005)

15.0
(0.012)

16.1

(0.005)

12,2
(0.010)

Dieldrin

4.3
(0.015)

2.7
(T)

1.6

(T)

2.4
(T)

Lindane

4.3

(0.003)

8.4

(0.008)

8.6

(0.003)

2.4
(0.018)

Toxaphene

30.4
(0,023)

1.3
(0.010)

1.1
(0.003)

12.2
(0.140)

BHC

8.7
(0.017)

2.7
(0.004)

4.8
(0.008)

—

Chlordane

8.7
(0.004)

2.2
(0.017)

6.5
(0.012)

2.4
(0.007)

Total Number
of Samples

23

226

186

41

Percent

w/Residues

78.3

53.5

39.8

41,5

TABLE 5. — Incidence of specific pesticide residues in corn
products

[T = <0.001 PPM; — = Not detected]

DDT

TDE
DDE

Dieldrin

Lindane

Toxaphene

Endrin

Chlordane

Percent of Samples Containing

Specific Pesticides and Average Level

(PPM) of Each Pesticide

Grain

5,7
(0.007)

1.2
(0.003)

2.9
(T)

4.4
(0.001)

2.6
(T)

0.1
(T)

Crude Oil

Total Number
of Samples

Percent
w/Residues

819
13.4

14.8
(0.067)

II. 1

(0.060)

11.1
(0.016)

11.1
(0.013)

Refined Oil

3.7
(0.080)

12.5
(0.038)

12.5
(0.002)

27
25.9

8
25.0

TABLE 6. — Incidence of specific residues in oleomargarine
[T = <0.001 PPM]

Percent Samples
Containing Pesticides

Average
Level (PPM)

DDT
TDE
DDE
BHC

18.9

5.7
7.5
3.8

0.026

0.002

0,001

T

Total Number of Samples: 53
Percent with Residues: 18.9

The chemical names of compounds mentioned in this paper are:

DDT l,l,l-trichloro-2,2-bis(p-chlorophenyl) ethane

TDE 1 ,l-dichloro-2,2-bis(p-chlorophenyl ) ethane

DDE l,l-dichloro-2,2-bis(p-chlorophenyl)cthylene

Dieldrin not less than 85% of l,2,3,4,10,10-hexachloro-6,

7-epoxy-l,4,4a,5,6,7,8,8a-octahydro-I,4-p«do-cJro-
5,8-dimethanonaphthaIene

Lindane 1,2,3.4,5,6-hexachlorocyclohexane, 99% or more

gamma isomer

Toxaphene chlorinated camphene containing 67% to 69%

chlorine

Endrin 1.2,3,4,10.10-hexachloro-6,7-£poxy-l,4,4a,5,6,7,8,8a-

octahydro-1.4-<>i!rfo-e'ndo-5,8-dimethanonaphthalene

BHC 1,2,3,4.5,6-hexachlorocyclohexane, mixed isomers

Chlordane 1 .2,4,5,6,7.8, 8-octachloro-3a.4,7,7a-tetrahydro-

4.7-methanoindane

Malathion diethyl mercaptosuccinate, 5-ester, with 0,0-di=

methyl phosphorodithioate

Aldrin not less than 95% of 1,2,3,4,10,10-hexachloro-l,

4,4a,5,8.8a-hexahydro-1.4-c«rfo-CA'o-5,8-dimethano=
naphthalene

Heptachlor epoxide 1 ,4,5,6,7, 8, 8-heptachloro-2,3-epoxy-3a,4.7,7a-
tetrahydro-4,7-methanoindan

Vol. 1, No. 4, March 1968

TABLE 7. — Summary — Average Icvch of chloriiuitcil pesticide residues in vegelahle oil seeds and products — fiscal

\etirs 1964 66

Sample

Raw

PKOilttI

[T = <0.001 PPM: — = Not detected)

Crude
Oil

Parts per Million

Meal
OR Cake

Refined
Oil

Oleo-
margarine

Total Diet
Composites '

DDT

Soybean

0.006

0.015

0.005

0.003

Cottonseed

0.154

0.077

0.028

o.o:4

Peanut

0.025

0.466

0.140

0.060

Corn

0.007

0.067

—

0.038

TOTAL

0.015

0.097

0.023

0.022

0.026

0.006

TOE

Soybean

T

0.006

T

Cottonseed

0.029

0.093

0.003

0.016

Peanut

T

0.128

0.026

0.007

Corn

0.003

0.060

—

0.002

TOTAL

0.001

0.072

0.005

0.010

0.002

0.008

DDE

Soybean

T

0.002

0.001

_

Cottonseed

0.005

0.012

0.005

0.010

Peanut

0.001

0.090

0.025

0.032

Corn

T

0.016

TOTAL

0.003

0.016

0.005

0.010

0.001

0.004

DIELDRIN

Soybean

0.002

0.013

T

T

Coiionsccd

0.015

T

T

T

Peanut

0.008

0.017

0.006

0.007

Corn

0.001

0.013

TOTAL

0.004

0.006

T

T

—

0.001

LINDANE

Soybean

T

T

0.001

Cottonseed

0.003

n.oos

0.003

0.018

Peanut

0.002

0.001

T

Com

T

TOTAL

T

0.005

0.003

0.01

—

T

BHC

TOXAPHENE

Soybean

0.004

0.024

T

Cottonseed

0.023

0.010

0.003

0.140

Peanut

0.006

0.008

Corn

TOTAL

0.017

0.015

0.002

0.078

—

—

ENDRIN

Soybean

0.008

0.013

T

Cottonseed

—

Peanut

T

0.008

Com

T

TOTAL

0.006

0.004

T

—

—

T

Soybean

Cotionsccd

Peanut

Corn

TOTAL

T
0.017
0.007

0.003

0.004
0.004
0.002

0.003

T

0.008
0.006

0.004

0.002
T

0.001

CHLORDANE

Soybean

I

_

T

Cottonseed

U.0U4

0.017

0.012

0.007

Peanut

Corn

0.080

TOTAL

T

0.017

0.006

0.004

—

—

■Includes salad dressings, salad oil, mayonnaise, shorlcninu. and pcanul hiillcr (74 composites, 6 64-4/67).

Pesticides Monitorinc; Journal

Acknowledgments

Recognition must be given to the chemists, too numerous
to mention as individuals, among the 18 FDA District
Laboratories responsible for these analyses and to R. K.
Dawson, Division of Program Operations, for his as-
sistance in processing the data.

LITERATURE CITED

(/.) Duggati, R. E. and Keilh Dawson. 1967. Pesticides —
A report on residues in food. FDA Papers 1:4.

(2) Diiggan, R. E. 1967. Pesticide residues in food. Con-
ference on Biological Effects of Pesticides in Mam-
malian Systems. N. Y. Acad, of Sci. May 1967.

(3) Duggan, R. E. and J. R. Wcatherwax. 1967. Dietary
intake of pesticide chemicals. Science 157:1007-1010.

(4) Duggan, R. E. 1967. Chlorinated pesticide residues
in fluid milk and other dairy products in the United
States. Pesticides Monit. J. l(3):2-8.

(5) Duggan, R. E. and F. J. McFarland. 1967. Residues
in food and feed — Assessments include raw food and
feed commodities, market basket items prepared for
consumption, meat samples taken at slaughter. Pesti-
cides Monit. J. l(l):l-5.

(6) Association of Official Analytical Chemists. Changes
in official methods of analysis. J. Ass. Offic. Anal.
Chem. 49:222(1966); ibid. 50:210-214(1967).

(7) Barry, Helen C, Joyce G. Hundley, and Lore:: Y.
Johnson. 1963 (Revised 1964. 1965). Pesticide an-
alytical manual. Food and Drug Admin., U. S. Dep.
Health, Educ, and Welfare, Washington, D. C.
20204.

(S) Johnson. L. Y. 1965. Collaborative study of a method
for multiple chlorinated pesticide residues in fatty
foods. J. Ass. Offic. Agr. Chem. 48:668-675.

(9) Krause, R. T. 1966. Collaborative study of a method
for multiple chlorinated pesticide residues in non-
fatty vegetables. J. Ass. Offic. Anal. Chem. 49:460-463.

(10) Gaul. J. 1966. Collaborative study of a method for
multiple chlorinated pesticide residues in leafy and
cole-type vegetables. J. Ass. Offic. Anal. Chem.
49:463-467.

(11) Wells. C. E. 1967. Vahdation study of a method for
pesticide residues in foods and animal feeds. J. Ass.
Offic. Anal. Chem. 50:1205-1215.

(12) V. S. Department of Agriculture. Agricultural Re-
search Service and Forest Service. 1967. Agriculture
Handbook No. 331. Suggested guide for the use of
insecticides to control insects affecting crops, live-
stock, households, stored products, forests, and forest
products.

Vol. 1, No. 4, March 1968

Investigation of Lead Residues on Growing Fruits and Vegetables^

Abram Kleinnian

ABSTRACT

An invesligation was inutic i>f the cxlent of lead residues on
crops grown near heavily traveled highways. Analyses are
presented of 132 samples of a variety of fruits and vege-
tables from four areas of the country. Lead residues arc
compared with distance from the highway, traffic load, and
the period of exposure to these conditions.

Possible contamination of growing food crops and crop-
growing areas by lead deposited from the atmosphere
has been a matter of concern for several years. Chow
and Johnstone (2) have estimated that an accumulation
of 10 mg of lead per square meter has been deposited
over the northern hemisphere since the advent of anti-
knock gasolines. Warren (4) has reported the presence
of lead in roadside vegetation. Cannon and Bowles (/)
have presented data, correlating the amount of lead
found in grasses with prevailing wind direction and
distance from highways.

The possible accumulation of excessive lead residues on
food crops grown near heavily traveled highways raised
the question of a possible hazard to public health. The
purpose of this investigation was to determine the extent
of such accumulation on growing crops.

Four FDA Held districts' participated in the investiga-
tion. A total of 132 samples of a variety of mature
fruits and vegetables were collected and analyzed for
lead. Samples collected ranged from 4 to 15 lb. All
samples were examined without washing or peeling. The
smaller samples were ground and mixed in entirety; the
larger samples were reduced to about I kg, then com-
posited, ground, and mixed. Appropriate aliquols (25
to 200 g) were analyzed by the otlicial A.O.A.C. di-
thizone spectrophotometric procedure (3), Reported

recoveries of added lead in recovery experiments ranged
from TO'vf to 100%. The results are summarized in
Table 1.

In attempting to relate lead content of the crop to
exposure to automobile exhaust, three parameters con-
sidered were distance from traffic, traffic load, and
period of exposure to the air.

The distance of the crop from the roadway was coded
as noted in footnote 1 of Table I . The three distance
codes and the number of samples in each code are shown
for each product. The distribution of samples by dis-
tance code is as follows:

Code No.

No. of Samples

1
2
3

49
32
51

' FootI .md Hruv; Adminislratlon. IJ. .S. Department of Hcalih,

callcin, and Welfare. I.os Annclcs. Calif. 9(1015.
- Atlanta, C incinnati, I.os Angeles. Philadelphia.

Fdu-

Tn reporting the second factor, traffic load, there was a
lack of uniformity among the districts. One district
arbitrarily classified the load as heavy, medium, or light
without defining the terms. The other districts named
the adjacent highway, indicating whether it was a U.S.
highway, turnpike. State highway, or local road. One
district reported the number of vehicles passing in a
10-minute period. This factor is shown in Table 1 as
heavy, medium, or light. U.S. highways and turnpikes
have been arbitrarily classified as heavy; State highways
as medium; and local roads as light. Where more than
one designation is shown, it means that the samples in
that group were distributed accordingly. A breakdown
of samples by traffic load factor for distance Codes 1
and 3 is shown in Table 2.

I he growth period (exposure period) was not deter-
mined during sample collection. The values shown in
Table I represent approximate periods which relate to

Pesticides Monitoring Journal

California, and may be subject to considerable variation
depending on local climate. They are submitted for
informational purposes only.

In an attempt to determine whether the above factors
influenced the lead burden on the crops, attention was
focused primarily on the distance from traffic. A statis-
tical comparison of the averages for distance Code 1
and distance Code 3 was performed, using the "t" test
for comparison of averages (5). These groups were
selected because they contained approximately equal
numbers of observations and showed a 10- to 100-fold
difference in distance from traffic. Possible sources of
bias would, of course, be present due to unequal dis-
tribution of different types of crops and unequal dis-
tribution of the traffic load factor between the two
groups. In addition, two observations (one in each
group) were outliers, namely, melons showing 0.71
ppm lead and collards showing 0.90 ppm lead. Both of
these outlying values were rejected on the basis of
Chauvenet's criterion (6).

The resulting distribution by traffic load is shown in
Table 2. Table 3 shows the distribution of the two
distance codes by fruit or vegetable group.

Although total balance for fruit or vegetable group and
traffic load factors is not perfect, it was felt that there
was some basis for a valid comparison of the two dis-
tance groups. The results of the "t" test for comparison
of the averages are shown in Table 4.

Discussion

The value of "t" calculated from the data exceeds, the
critical value at the 1 % level of significance for the
"one-tailed" distribution of this statistic. The "one-
tailed" distribution of "t" would be the proper one to
use if we seek the answer to the question, "Is the lead
content of crops growing adjacent to traffic greater than
that of crops at further distances?" The data suggest that
such a difference may exist. However, this conclusion
must be viewed in the light that other possible sources
of lead have been ignored, i.e., pesticides that contain
lead, lead accumulation in the soil, and the possible
sources of bias mentioned earlier.

LITERATURE CITED

(/) Cannon. H. L. and J. M. Bowles. 1962. Contamina-
tion of vegetation by tetraethyl lead. Science 137:765-
766.

(2) Chow. T. J. and M. S. Johnstone. 1965. Lead isotopes
in gasoline and aerosols of Los Angeles Basin, Calif.
Science 147:502-503.

(.?) Association of Official Agricultural Chemists. 1965.
Official methods of analysis, 10th ed.. Section 24.041,
et seq.

(4) Warren, H. V. 1961. Some aspects of the relationship
between health and geology. Canad. J. Public Health
52:157.

(5) Youden, W. J. 1951. Statistical methods for chemists.
John Wiley & Sons, Inc., New York.

(6) Young. H. D. 1962. Statistical treatment of experi-
mental data. McGraw-Hill Book Co., New York.

TABLE 1. — Lead content of fruits and vegetables correlated with distance from traffic and traffic load

Distance
FROM Traffic >

Number of
Samples

Traffic
Load -

Growth

Period

(Weeks) "

Lead Content (PPM)

Product

Averagf

Range

Grapefruit

1
2
3

2
1
8

M

H

H,M,L,

13
13
13

0.08
0.03
0.02

0.06—0.09

0.01—0.05

Oranges

1
2
3

7
4
2

M,L,
H.L
M

13
13
13

0.09
0.08
0.12

0.03—0.22
0.03—0.16
0.11—0.13

Lemons

1

2
3

4
2
1

H,M
H,L

M

12-16
12-16

12-16

0.15
0.18
0.01

0.13—0.17
0.11—0.25

Cantaloupe

or Honey Dew

1

2

9
1

L
M

7-8
7-8

0.16
0.02

0.01—0.71

Strawberries

1
3

3

5

H,M,L
H,L

22
22

0.10
0.15

0.04—0.14
0.07—0.16

Peaches

3

1

M

—

0.00

—

Collards

1
2
3

2
4

1

H,M
H,M

M

12
12
12

0.29
0.21
0.90

0.28—0.30
0.15—0.30

Lettuce

1

2
3

5

1
5

M,L

M

H.M.L

12-14
12-14
12-14

0.10
0.26
0.06

0.05—0.22
0.03—0.07

Endive

3

3

M,L

12

0.05

0.02—0,10

Spinach

2

2

L

28

0.27

0.19—0.35

Broccoli

1

2
3

1
5
4

M
H,M,L
H,M,L,

8-12
8-12
8-12

0.02
0.30
0.02

0.05—0.65
0.00—0.03

Vol. 1, No. 4, March 1968

TABLE I.- I iiui iimti-nt of fruits and vegetables correlated with distance from traffic and traffic load — Continued

^ Distance from irafTic coded as follows: 1 = 0 lo 25 yds; 2 = 25 to 250 yds; 3 = above 250 yds.

= H = Heavy; M = Medium; L =: Light.

■TTiese are general estimates related primarily to California; they represent time of total exposure to air.

DisrANcc
1 K(iM Traffic '

Number of
Samples

Traffic
Load -

Growth

Period

(Weeks) »

Lead Content (PPM)

PHODUCT

Average

Range

Cabbane

1

2
3

4
1
3

M.L

H

H,M,L

8-27
8-27
8-27

0.03
0.00
0.02

0.00—0.04
0.00—0.04

Turnip Greens

3

1

H

18

0.31

—

Rape

3

1

H

13

0.25

—

Tomatoes

1
2
3

2
2

7

H

M
H.M.L

12

12
12

0.03
0.05
0.05

0.00—0.05
0.04 — 0.05
0.01—0.07

Squash

3

1

M

7-8

0.00

—

Pole Beans

2

1

L

12

0.12

—

Green Beans

3

1

M

12

0.00

—

Potaloes

1
3

2
1

M
M

10
10

0.02
0.01

0.01—0.02

Carrots

1
2
3

5
2
3

M,L
M.L
M,L

12
12
12

0.09
0.03
0.03

0.03—0.16
0.02—0.03
0.00—0.05

Radishes

2

1

L

8

0.06

-

Celery

1
2
3

2
4
2

M
M
M

26-28
26-28
26-28

0.14
0.12
0.16

0.09—0.18
0.07—0.15
0.05—0.26

Cauliflower

2
3

1
1

L
H

8-12
8-12

0.03
0.03

Asparagus

1

1

M

16

0.00

—

TABLE 2. — Distrihulion of scuuplcs by traffic load for
distance Codes I and 3 '

TABLE 4. — Comparison of averages for
1 and 3

distance Codes

No. of Samples

Traffic Load

Distance Code 1

Distance Code 3

Heavy

Medium
Light

5
21
22

18
18
14

Distance Code 1 Distance Code 3

* Does not include two outlying values rejected on basis of Chauvenet's
criterion.

TABLE 3. — Distribution of samples by fruit or vegetable
group for distance Coues I and 3 '

Fruit or Vegetable

No. OF Samples

Group

Distance Code 1

Distance Code 3

Citrus

13

11

Melons

0

Other Fruits

3

6

Leafy Vegetables

12

17

Vine Vegetables

9

Root Vegetables

4

Other Vegetables

3

Average (ppm Pb)
Variance (S-)

Standard deviation of

difference between averages

F (comparison of variances)

The Sludsdcal Test
"t" (test statistic)
Degrees of freedom

Critical value of "t" at
1% level of significance

for 60 D.F. 2.39

("one-tailed" distribution)

0.0910

0.0562

0.00510

0.0139
1.17

2.51
96

0.00434

' Docs not include (wo outlying values rejected on basis of Chauvenet's
criterion.

10

Pesticides Monitoring Journal

Pesticide Residues in Total Diet Samples (III)

R. J. Martin' and R. E. Duggan"

ABSTRACT

Pesticide residue levels detected in recidy-to-eiit foods re-
mained at low levels during the third year of the total diet
study. Samples were collected from 30 markets in 29 dif-
ferent cities.

Population of cities ranged from less than 50.000 to 1,000.-
000 or more. A verages and ranges of pesticides commonly
found are reported for the period June 1966 — April 1967
hy region and food class. Pesticides found infrequently al.fo
are reported for this period by region and food cla.is.

The study of pesticide residues in ready-to-eat foods,
conducted by the Food and Drug Administration from
June 1964 through April 1966, has been described in
earlier reports (/). This report covers the period June
1966 through April 1967. Tabular data are included
comparable to that reported for the previous years. No
changes were made in the sampling and compositing
procedures given in the "Food and Feed Section" of the
Pesticides Monitoring Journal (2) which describes the
National Pesticide Monitoring Program. Earlier reports
(3,4) discuss data collected from June 1964 through
April 1965 and June 1965 through April 1966, re-
spectively.

Samples were collected from 30 markets in 29 different
cities. Population of cities ranged from less than 50,000
to 1.000,000 or more. The samples were analyzed for
the presence of chlorinated hydrocarbons, organic

^ Field Scientific Coordination Branch. Bureau of Science. Food and

Drug Administration, Washington. D.C. 20204,
- Office of Associate Commissioner for Compliance, Food and Drug

Administration, U. S. Department of Health, Education, and Welfare,

Washmgton, D.C. 20204.

phosphates, chloropheno.xy acids, bromides, arsenic,
amitrole (3-amino-l,2,4-triazole), carbarbyl (Sevin®),
and dithiocarbamate residues.

Quantitative values reported for both chlorinated and
organic phosphorus compounds were obtained by either
electron capture or thermionic gas-liquid chromatog-
raphy. Confirmation was made by thin layer chro-
matography and/or microcoulometric gas-liquid chro-
matography. This procedure determines chlorinated
compounds at a sensitivity (quantitative) of 0.003 ppm
and organic phosphorus compounds at 0.05 ppm. Each
composite was also tested for chlorophenoxy acids and
esters at a sensitivity of 0.02 ppm: for amitrole at a
sensitivity of 0.05 ppm; for dithiocarbamates, calculated
as zineb (zinc ethylene-l, 2-bisdithiocarbamate) at a
sensitivity of 0.2 ppm: for carbaryl at a sensitivity of
0.2 ppm: for bromides at a sensitivity of 0.5 ppm: and
for arsenic as As^O;, at a sensitivity of 0.1 ppm.

All methods used in these studies are described in the
FDA Pesticide Analytical Manual. Vol. 1 and 11 (5).
Recoveries of specific pesticide chemicals vary within
product classes, usually within a range of 85% to
115% at these levels. No correction was made for
recovery.

RESULTS

A total of 997 residues were detected during this current
reporting period. There was no significant change in
the levels, frequency, or types of residues found from
those in the past.

Twenty-nine different residues were found in the samples
in 1967. The frequency of the residues is shown in

Vol. 1, No. 4, March 1968

11

Table 1. The most common residues, maximum levels of
(hose residues, and residues reported less frequently are
discussed below for each class.

DAIRY PRODUCTS: Thirteen chlorinated organic
pesticides in varying combinations were detected in 27
of 30 composites. The most common, and their max-
imum values on a fat basis, were: DDE (0..10 ppm);
DDT (0.14 ppm); dicldrin (0.08 ppm); heptachlor
epoxide (0.0.1 ppm); TDE (O.IS ppm); and BHC (0.05
ppm). Also present were aldrin, heptachlor, lindane,
methoxychlor, 2,4,5-T, 2,4-D, PCP, Kelthane^, and
arsenic (.As-.O,). Bromides were found (0.5 ppm to
21.3 ppm) in 28 of 30 composites.

MEAT, FISH, AND POULTRY: Ten chlorinated or-
ganic pesticides were present in varying quantities in 29
of 30 composites. DDT, DDE, TDE, heptachlor epoxide,
dieldrin, and BHC were the most common, with max-
imum values of 0.882 ppm, 0.755 ppm, 0.69 ppm, 0.105
ppm, 0.120 ppm, and 0.06 ppm, respectively, on a fat
basis. Aldrin, lindane, PCP, and phorate were also
present. Bromides were detected (0.8 to 47.2 ppm) in
27 of 30 composites; Arsenic (AsjO.) was detected 9
times at values ranging from 0.1 to 0.5 ppm; and 2,4,5-T
was detected in 1 composite.

GRAIN AND CEREAL PRODUCTS: Nine chlorinated
organic pesticides were found in 28 of 30 composites
with the most common being lindane, DDT, and dieldrin,
at maximum values of 0.171 ppm, 0.02 ppm, and 0.011
ppm. respectively. DDE, BHC, heptachlor epoxide,
aldrin, PCP, and TDE also were present. Bromides were
detected (0.5 ppm to 47 ppm) in 28 of 30 composites.
Eight composites contained malalhion, with a maximum
value of 0.13 ppm. Arsenic (As^O^t) and carbaryl also
were present.

POT .A TOES: The most common pesticides found were
DJjfl and DDE at maximum values of 0.03 ppm and
0.02 ppm, respectively. These 2 were detected in 12 of
30 composites. Other chlorin-'id organic pesticides
present were dieldrin, CIPC, lindane, TDE, and PCP.
Endrin was detected in 5 of 30 composites at a max-
imum value of 0.01 ppm. Bromides were found in 25 of
30 composites. Values ranged from 0.3 ppm to 57.2
ppm.

LEAFY VEGETABLES: DDT, DDE, and TDE with
maximum values of 0.058 ppm, 0.02 ppm. and 0.045
ppm, respectively, were detected in 22 of 30 composites.
Aldrin, BHC, chlordane, dieldrin. endrin, and lindane
were also present. Paralhion was found in 3 of 30 com-
posites, with a maximum value of 0.04 ppm. Methyl
parathion, cndosulfan, and arsenic (As^O^,) were each

detected 1 time. Dithlocarbamates (calculated as zineb)
were found twice at 0.44 and 0.8 ppm levels. Bromides
were detected in 24 of the 30 composites.

LEGUME VEGETABLES: DDE, TDE, and DDT were
found in 8 of 30 composites, with maximum values of
0.01 ppm, 0.05 ppm, and 0.062 ppm, respectively.
Aldrin, chlordane, and lindane were also present. Arsenic
(AsjOj) was detected twice, with a maximum value of
0.18 ppm. Bromides were found (0.5 ppm to 19 ppm)
in 22 of the 30 composites.

ROOT VEGETABLES: TDE, DDT, and DDE were
detected in 8 of the 30 composites at maximum values
of 0.02 ppm, 0.04 ppm, and 0.01 ppm, respectively.
Endrin was detected in I composite. Carbaryl and di-
thiocarbamates (calculated as zineb) were each detected
I time with values of 0.05 ppm and 0.32 ppm, respec-
tively. Bromides were detected (0.3 ppm to 20.5 ppm)
in 26 of the 30 composites. Arsenic (AsnO:)) was de-
tected 3 times with a maximum value of 0.16 ppm.

GARDEN FRUITS: A total of 8 chlorinated organic
residues were detected in 27 of the 30 composites. DDT,
TDE, and DDE were the most common with maximum
values of 0.19 ppm, 0.02 ppm, and 0.04 ppm, re-
spectively. Dieldrin, lindane, aldrin, heptachlor epoxide,
and TCNB also were present. Diazinon, carbaryl, and
parathion were all detected 1 time with values of 0.003
ppm, 0.10 ppm, and 0.014 ppm, respectively. Bromides
were detected (1.1 ppm to 12 ppm) in 28 of the 30
composites.

FRUITS: Ten chlorinated organic residues were found
in 25 of the 30 composites. DDT, DDE, Kelthane®,
TDE, and aldrin were found most frequently with max-
imum values of 0.09 ppm, 0.04 ppm, 0.23 ppm, 0.025
ppm, and 0.015 ppm, respectively. Methoxychlor, lin-
dane, BHC, heptachlor epoxide, and dieldrin were also
present. Ethion was detected 3 times with a maximum
value of 0.054 ppm. Arsenic (AsoO-i) occurred 3 times
varying from 0.1 to 0.2 ppm. Carbaryl was detected 1
time, at a level of 0.22 ppm. Bromides were detected
24 times (0.6 ppm to 34.1 ppm) in 30 composites.

OILS, FATS. AND SHORTENING: A total of 7
chlorinated organic residues were detected in 21 of the
30 composites. DDE, DDT, and TDE were the most
common, with maximum values of 0.03 ppm, 0.023
ppm, and 0.04 ppm, respectively. Dieldrin, BHC, lin-
dane, and PCP were also present. Bromides were de-
tected (0.7 ppm to 49.1 ppm) in 25 of 30 composites.
Malathion was detected 4 times, ranging from trace to
0.062 ppm. Diazinon and ethion were each detected 1
time. Arsenic (As^O.j) occurred twice at a 0.1 ppm
level.

12

Pesticides Monitoring Journal

TABLE 1. — Number of composites where pesticide residues were found nnd ranges in the

amounts (June 1966 - April 1967)

Pesticide

No.

COMPOSITES

WITH

RESIDUE

No. OF POSITIVE

COMPOSITES WITH

RESIDUES BELOW

SENSITIVITY LEVEL ^

Range at and
above sensi-
tivity level

(PPM)

BROMIDES

301

3

0.5-57.2

DDT

I, l,l-trichloro-2.2-bis(p-chlorophenyl) ethane

145

6

0.003-0.882

DDE

l,I-dichloro-2,2-bis(p-chlorophenyl) ethylene

123

13

O.003-O.755

TDE

l,l-dichloro-2,2-bis(p-chlorophenyl) ethane

112

13

0.003-0.69

DIELDRIN

not less than 85% of l,2,3,4,10,I0-hexachloro-6,7-epoxy-l,4,4a,5,6,7.8,8a-octahydro-
l,4-endo-«xo-5,8-dimethanonaphthalene

58

7

0.003-0.12

LINDANE

1,2,3.4,5,6-hexachlorocyclohexane, 99% or more gamma isomer

49

14

0.003-0.374

ARSENIC (AsiOa)

33

8

0.1-0.40

BHC

1,2,3,4,5,6-hexachlorocycIohexane, mixed isomers

34

4

0.003-0.06

HEPTACHLOR EPOXIDE

1,4,5,6,7,8, 8-heptachloro-2,3-epoxy-3a,4,7,7a-tetrahydro-4,7-methanoindan

33

4

0.005-0.17

KELTHANE®

4,4'-dichloro-a- ( trichloromethyl ) benzhydrol

20

0

0.019-0.23

ALDRIN

not less than 95% of 1,2,3,4, 10,10-hexachloro-l,4,4a,5,8,8a-hexahydro-l,4-?ndo-exo-
5,8-dimethanonaphthalene

14

3

0.003-0.03

MALATHION

diethyl mercaptosuccinate, S-eslet with 0.0-dimethyl phosphorodithioate
PCP

pentachlorophenol

14
11

4
2

0.05-0.19
0.007-0.043

ENDRIN

1,2, 3,4, 10, 10-hexachloro-6.7-epoxy-l, 4,4a. 5.6,7,8, 8a-octahydro-l,4-<'ndo-e'n<io-
5,8-dimethanonaphthaIene

7

2

0.003-0.05

2,4-D

2,4-dichlorophenoxyacetic acid

6

2

0.027-0.08

PARATHION

0,0-diethyl 0-p-nitrophenyl phosphorothioate

5

4

0.093

CARBARYL

1-naphthyl methylcarbamate

4

2

0.22-0.34

ETHION

0,0,0' ,0'-tetraethyl-5„y'-methylene bisphosphorodithioate

4

2

0.0544).25

METHOXYCHLOR

1,1,1 -trichloro-2,2-bis ( p-methoxyphenyl ) ethane

3

1

0.004-0.09

DITHIOCARBAMATES

3

0

0.32-0.8

DIAZINON

0,0-diethyl 0-(2-isopropyl-6-methyl-4-pyrimidinyl ) phosphorothioate

2

2

CHLORDANE

1,2,4,5,6,7,8, 8-octachloro-3a,4,7,7a-tetrahydro-4,7-methanoindane

2

0

0.005-0.02

HEPTACHLOR

1, 4,5,6,7,8, 8-heptachloro-3a,4,7,7a-telrahydro-4,7-methanoindene

1

0

0.02

CIPC

isopropyl A'-(3-chlorophenyl) carbamate

1

0

0.11

2,4,5-T

2,4,5-trichlorophenoxyacetic acid

2

1

0.19

TCNB

I,2,4,5-tetrachloro-3-nitrobenzene

1

0

0.004

ENDOSULFAN

6,7,8,9,10,I0-hexachloro-l,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-ben2odioxathiepin
3-oxide

1

0

0.003

METHYL PARATHION

0,0-dimethyl 0-p-nitropheny! phosphorothioate

1

1

PHORATE

0,0-diethyl J-Cethylthio) methyl phosphorodithioate

1

1

Pesticide chemicals capable of being detected by the specified analytical methodology
when they are present at concentrations below the sensitivity level.

may be confirmed qualitatively but are not quantifiable

Vol. 1, No. 4, March 1968

13

SUGARS AND ADJIINCTS: Fight chlorinated organic
residues were detected m 14 of the 30 composites. 2.4-D
was found in 5 composites with a maximum value of
0 08 ppm: DDK and DDT were each found 3 times
with maximum values of 0.02 ppm and 0.002 ppm, re-
spectively. Kelthane®. aldrin, lindane, TDE, and PCP
were also present. Malathion was detected in 1 com-
posite at 0.07 ppm. Arsenic (AsoO.,) was detected 4
times with a maximimi value of 0.15 ppm. Bromides
were detected (1.1 ppm to 42.9 ppm) in 25 of 30
composites.

BEVERAGES: Iwo chlorinated organic residues were
detected in 2 composites. PCP at 0.021 ppm was found
in 1 composite and a trace of lindane was reported in
the other. Bromides were found in 19 composites. Con-
centrations ranged from 0.5 ppm to 14.7 ppm. Malathion
was detected 1 time at a level of 0.19 ppm. Arsenic
(As^.O-,) occurred I time at a level of 0.25 ppm.

Values include naturally occurring bromides as well as
residues from pesticide treatment. The quantitative
sensitivity limits were established for the study in 299
of the 360 composite samples. This incidence is 83.1%
and does not differ significantly from the 76.8% in-
cidence found in the 1965-1966 results. A total of 4.2%^
of the residues excectled 25 ppm while an incidence of
3.8% was reported for 1964-1965,

The data obtained during the third year of the study are
reported in more detail in Table 2a, where the findings
are arranged by food class and region. Similar informa-
tion is given in Table 2b for pesticide residues found
infrequently (less than five detections per commodity
class). The data are reported in the same format used
for the earlier period (3) for ease of comparison. Trace
amounts, <0()01 ppm, are not included in the averages.
Where no average value is given, the results of individual
composites are shown. In these tabulations, as in the
earlier report, the bromide and arsenic values are re-
ported on an "as is" basis for three food classes: Dairy
Products (I); Meat, Fish, and Poultry (II): and Oils,
Fats, and Shortening (X), even though the earlier tab-
ulations indicated a "fat basis."

Disct4Ssion

The pre^encc of chlorinated organic residues was con-
firmed in 224 of the 360 composites examined (62.3% )
for this class of chemicals. This percentage of incidence
is not significantly different from the 1965-66 data
(53.8%). Organic phosphorus compounds were found
in 25 composites; 27 detections were reported for
1965-66. Relatively few carbaryl values have been re-
ported during the entire study, and the majority of these

were reported by Kansas City; however, Boston, Los
Angeles, and Baltimore have also reported positive
findings. Carbaryl was detected (0.05-0.34 ppm) in 4
composite samples for the current year. Each composite
was analyzed by thin layer chromatography. Positive
results were confirmed and quantitated spectrophoto-
metrically (5). In considering the carbaryl values, it
must be recognized that at the lower limits of sensitivity
of the method, 0.1 to 0.2 ppm, the accuracy of the
method is reduced.

For example, carbaryl and other chemicals found in-
frequently in less than 1 % of the composites, cannot
be considered as a regular component of residues in the
diet.

Chlorophenoxy acids were found in 8 composites for
the current year; 13 residues were reported for 1965-66.
Dithiocarbamates (calculated as zineb) were found in
3 composites. No detections were reported for 1965-66,
while 4 values were reported for 1964-65.

Samples have been analyzed for the presence of amitrole
since initiation of the program, but no residues have
been detected. Levels and kinds of residues for this
period remain in the same order of magnitude as those
reported in the earlier studies. The frequency has not
changed significantly as a whole or within each food
class.

On the basis of these data, we can reasonably conclude
that there is no significant difference in the dietary
intake among the three reporting periods of this study.

A cknowledgment

The authors gratefully acknowledge the analytical work
from the FDA Laboratories in Baltimore, Md.; Boston,
Mass.; Kansas City, Mo.; Los Angeles, Calif.; and
Minneapolis, Minn,

LITERATURE CITED

(/) nuf-Kiin. R. E.. It. C. Barry, and L. Y. Johnson. 1966.
Pesticide residues in total diet samples. Science 151 : 101-
104.

(2) Dni^i^an, R. E. und F. J. McFarUinJ. 1967. Assess-
ments include raw food and feed commodities, market
basket items prepared for consumption, meat samples
taken at slaughter. Pesticides Monit. J. 1(1): I.

(.?) DiifiU'iii. R E.. H. C. Harry, and L. Y. Johnson. 1967.
Pesticide residues in total diet samples (11). Pesticides
Monit. J. I(2):2-I2.

[4) Diijigan. R. E. and J. R. Weatherwa.x. 1967. Dietary
intake of pesticide chemicals. Science 157:1006-1007.

(5) Barry. H. C, J. G. Hundley, and L. Y. John.son. Pesti-
cide analytical manual. Vol. I and II., Food and Drug
Admin., U. S. Dcp. Health, Educ, and Welfare.

14

Pesticides Monitoring Journal

TABLE 2a. — Levels of pesticide residues commonly found — by food class and region (June 1966-April 1967)

[T = Trace<0.001 ppm]

Pesticide

Boston

Kansas City

Los Angeles

Baltimore

Minneapolis

I. DAIRY PRODUCTS (8-13% fat) i
Residues in Parts Per Million — Fat Basis

DDT

Average

Positive Composites

Number

Range

0.033

4
0.023-0.05

0.023

5
0.015-0.03

0.109

3
0.056-0.14

0.029

4
0.012-0.050

0.070

5
T-0.11

DDE
Average
Positive Composites

Number

Range

0.029

5
0.025-0.038

0.017

5
0.006-0.042

0.203

5
0.019-0.30

0.021

4
0.01-0.026

0.03

5
0.011-0.050

TDE

Average

Positive Composites

Number

Range

0.018

2
0.017-0.020

O.OU

4
0.006-0.02

0.013

3
0.06-0.18

0.027

5
0.009-O.04

0.026

3
0.012-0.04

DIELDRIN

Average

Positive Composites

Number

Range

0.038

3
0.013-0.08

0.027

5
0.008-0.05

0.025

5
0.008-0.051

0.028

3

0.02-0.035

0.035

3
0.007-0.080

HEPTACHLQR EPOXIDE

Average

Positive Composites

Number

Range

0.015

4
0.008-0.020

0.012

3
T-0.012

0.012

2
0.007-0.017

0.01

2
0.01-0.013

0.013

5
0.005-0.030

BHC

Average

Positive Composites

Number

Range

0.025

4

0.015-0.050

0.029

3
0.020-0.039

0

0

0.027

5

0.016-0.040

TOTAL BROMIDES
Average
Positive Composites

Number

Range

9.2

6
0.8-21.3

6.3

6

1.3-8.4

3.6

6
1.2-9.0

5.6

6

0.7-10.0

4.7

4
0.5-14.3

II. MEAT, FISH, AND POULTRY (17-23% fat) ■
Residues in Parts Per Million — Fat Basis

DDT

Average

Positive Composites

Number

Range

0.370

5
0.12-0.882

0.177

6

0.051-0.40

0.139

6

0.095-0.20

0.107

5
0.01-0.160

0.152

5
0.081-0.221

DDE

Average

Positive Composites

Number

Range

0.266

6

0.07-0.459

0.123

6
0.03-0.166

0.341

6

0.08-0.755

0.066

5
0.011-0.110

0.064

5
0.04-0.100

TDE

Average

Positive Composites

Number

Range

0.251

6
0.04-0.69

0.105

6

0.022-0.28

0.089

6

0.05-0.15

0.052

6

0.004-0.10

0.051

4
0.014-0.09

DIELDRIN

Average

Positive Composites

Number

Range

1
0.061

0.059

6

0.016-0.120

0.013

4
0.008-0.02

0.015

2
0.01-0.02

0.023

3
0.008-0.04

HEPTACHLQR EPOXIDE

Average

Positive Composites

Number

Range

0.068

3
0.05-0.105

0.019

4
0.009-0.033

0.033

2
0.006-0.06

0

O.OU

4

0.006-0.02

BHC

Average

Positive Composites

Number

Range

0.040

2
0.020-0.06

0.021

6
0.01(M).030

0.025

3
0.006^.05

0

O.OU

4
0.007-0.02

Vol. 1, No. 4, March 1968

15

TABLE 2a. — Levels of pesticide residues commonly found — by food class and region (June 1966 - April 1967) — Continued

Pesticide

16

Boston

Kansas City

Los Angeles

Baltimore

II (a). MFAT. FISH. AND POULTRY (I7-23';f fat) '—(Continued)
Residues in Parts Per Million — Fat Basis

III. GRAIN AND CEREAL'
Residues in Parts Per Million

IV. POTATOES'
Residues in Parts Per Million

V. LEAFY VEGETABLES'
Residues in Parts Per Million

Minneapolis

ARSENIC (ASiO.)
Average
Posilivr Composites

Number

Range

0.23

4
0.1-0.33

1

0.5

0.22

4
0.1-0.38

0

0

TOTAL BROMIDES
Average
Pnutive Composites

Number
Range

11.5

5
6.9-20.2

6.9

6

2.9-15.0

5.1

5
1.3-14.0

20.6

6

7.9^7.2

4.2

5

0.8-5.4

DDT

Average

0.015

0.007

0.005

0.0O9

Positive Composites

Number

3

2

4

4

0

Range

T-0.02

0.006-0.007

0.003-0.007

0.0O5-0.012

DIEIDRIN

Average

0.005

0.006

0.006

Positive Composites

Number

2

4

2

1

0

Range

T-O.005

T-0.008

0.005-0.006

0.011

LINDANE

Average

0,004

0.060

0.003

0.009

Positive Composites

Number

5

5

6

1

5

Range

0.003-0.005

0.003-0.171

T-0.005

0.003

0.006-0.011

MALATHION

Average

0.038

0.115

Positive Composites

Number

0

4

1

2

1

Range

0.015-0.05

0.009

0.099-0.13

0.063

TOTAL BROMIDES

Average

22.5

19.3

9.9

18.0

14.8

Positive Composites

Number

6

6

6

6

4

Range

15.7-30.2

5.3^7.0

5.1-15.4

0.5-40.2

10.9-17.6

DDT

Average

Positi\e Composites
Number

0.005
4

0

0.003
2

1

1

Range

0.004-0.01

0.003

0.005

0.03

DDE

Average

Positive Composites
Number

0.004
4

1

0.004

2

1

1

Range

T-0.(K)4

0.006

T-0.004

0.009

0.02

ENDRIN

Average

Positive Composites
Number

2

0

0.003
2

0

1

Range

T-0.01

0.002-0.004

0.005

TOTAL BROMIDES

Average

Positive Composites
Number

8.1
6

6.6

5

7.7

4

19.2
6

4.5
4

Range

1.1-17.6

1.9-22

3.6-14.3

5.7-57.2

0.3-9.5

DDT

Average

0.015

0,019

0.039

0.025

0.033

Positive Composites

Number

6

3

5

4

2

Range

too;

0.011-0.028

0.005-0.058

0.003-0.056

0.025-0.04

Pesticides Monitoring Journal

TABLE 2a. — Levels of pesticide residues commonly found — by food class and region (June 1966 - April 1967) — Continued

Kansas City

Los Angeles

V. LEAFY VEGETABLES '—(Continued)
Residues in Paris Per Million

VL LEGUME VEGETABLES i
Residues in Parts Per Million

VIL ROOT VEGETABLES'
Residues in Parts Per Million

VIIL GARDEN FRUITS'
Residues in Parts Per Million

Baltimore

Minneapolis

DDE

Average

0.011

0.008

Positive Composites

Number

2

0

5

1

1

Range

0.002-0.02

0.007-0.009

0.004

0.01

TDE

Average

0.010

0.015

Positive Composites

Number

2

1

5

1

1

Range

0.009-0.011

T

0.003-0.045

0.006

0.01

TOTAL BROMIDES

Average

5.6

3.7

1.4

4.4

1.5

Positive Composites

Number

6

6

3

5

4

Range

2.4-9.5

0.8-10

0.9-2.6

1-6.7

0.7-1.9

DDT

Average

Positive Composites

Number

Range

1
0.031

0

0.033

2
0.003-0.062

1
0.04

0

TDE

Average

Positive Composites

Number

Range

0

0.004

2
T-0.004

0.029

3
0.006-0.05

0

0

DDE

Average

Positive Composites

Number

Range

1
T

2
T-0.01

0.004

3
0.003-0.005

0

0

TOTAL BROMIDES
Average
Positive Composites

Number

Range

7.6

6

1.6-13.8

5.4

4
0.5-19

3.5

4
1.1-7.8

5.2

4
1.7-9.6

1.8

4
0.8-3.0

DDT

Average

0.02

0.022

Positive Composites

Number

0

2

0

2

0

Range

0.01-0.02

0.003-0.04

DDE

Average

0.003

0.007

Positive Composites

Number

0

1

2

3

1

Range

0.01

0.003

0.005-0.009

0.004

TDE

Average

0.005

Positive Composites

Number

0

1

2

2

1

Range

0.02

0.005

0.004-0.01

0.005

TOTAL BROMIDES

Average

5.7

3.3

3.2

5.4

1.8

Positive Composites

Number

6

6

4

6

4

Range

0.5-20.5

0.9-8.5

0.4-8.8

0.3-13.3

0.9-2.7

DDT

Average

Positive Composites

Number

Range

0.082

0.059-0.12

0.038

0.016-0.054

0.059

6

0.006-0.19

0.018-0.092

0.021

0.007-0.035

Vol. 1, No. 4, March 1968

17

TABLE 2a. — Levels of pesticide residues commonly found — by food class and region (June 1966 - April 1967) — Continued

Pesticide

Boston

Kansas City

Los Angeles

Baltimore

Minneapolis

Mil GARDEN FRUITS '—(Continued)
Residues in Parts Per Million

DDE
Average
Positive Composites

Number

Range

0

0.03

2
0.02-0.04

0.018

3
0.003-0.03

0.006

2
0.003-0.01

0.017

3
T-0.03

TDE

Average

Fosttixe Composites

Number

Range

0

0.033

5
0.008-0.089

0.008

4

0.004-0.013

0.008

3
0.002-0.02

0.008

3
T-0.01

DIELDRIN
Average
Pontine Composites

Number

Range

1

0.003

1
0.003

0.002

2
0.001-0.003

0

1
0.011

LINDANE
Average
Positive Composites

Number
Range

0.002

3
T-0.0O2

0.027

2
0.0O4-O.049

1
0.002

0

0

TOTAL BROMIDES

Average

Positive Composites

Number

Range

6.4

6
1.8-11.7

2.4

6

1.1-2.9

4.7

6

1.1-12.0

5.2

6

2.0-9.4

1.7

4
1,1-3.1

IX. FRUITS »
Residues in Parts Per Million

DDT

Average

Positive Composites

Number

Range

0.035

5
0.005-0.08

0.01

2
O.OI-O.Oll

0.059

3
T-0.09

0.01

2
0.01

0.009

2
0.009

DDE

Average

Positive Composites

Number

Range

O.0O5

3

T-0.005

0.04

2
T-0.04

0.003

5
0.001-0.007

0

0.005

2
0.003-0.006

TDE

Average

Positive Composites

Number

Range

0.013

3
T-0.025

0.008

2
T-0.008

0.004

3

0.003-0.005

0.013

2
0.006-0.02

0.008

2
0.005-0.01

ALDRIN
Average

Positive Composites
Number
Range

1
0.008

0

0

0.003

2
0.002-0.003

0.008

3
0.004-0.015

K.ELTHANE®

Average

Positive Composites

Number

Range

0.121

5
0.06-0.23

0.067

5
0.019-0.133

0.046

5
0.022-0.10

1
0.02

0.068

2
0.035-0.10

TOTAL BROMIDES
Average
Positive Composites

Number

Range

3.1

5
0.8-5.5

4.3

6

0.9-17.0

5.7

4
0.6-13.0

11.9

6

3.7-34.1

4.4

3
2.1-5.8

X. OILS, FATS, AND SHORTENING i
Residues in Parts Per Million

UUl

Average

0.009

0.007

0.013

Positive Composites

Number

1

0

2

4

4

Range

0.021

0.007-0.01

0.006-0.01

T-0.023

DDE

Average

0.02

0.006

0 011

Positive Composites

Number

I

1

2

4

5

Range

T

0.005

0.01-0.03

T-0.01

T-0.02

18

Pesticides Monitoring Journal

TABLE 2a. — Levels of pesticide residues commonly found — by food class and region (June 1966 - April 1967) — Continued

Pesticide

Boston

Kansas City

Los Angeles

Baltimore

X (a). OILS, FATS, AND SHORTENING i— (Continued)
Residues in Parts Per Million

XI. SUGARS AND ADJUNCTS'
Residues in Parts Per Million

XII. BEVERAGES 1
Residues in Parts Per Million

Minneapolis

TDE

Average

Positive Composites
Number

1

1

0.023
2

0.003
2

0.016
4

Range

0.04

0.01

0.015-0.03

0.002-0.004

T-0.02

TOTAL BROMIDES

Average

Posiii\e Composites
Number

5.7
6

6.8
6

4.6

4

28.9
5

6.7

4

Range

0.7-10.9

1.9-24.0

3.7-6.6

2.7-49.1

3.5-11.2

2, 4-D

Average

Positive Composites

Number

Range

0

1
0.01

1

0.08

0

0.033

3

0.016-0.05

TOTAL BROMIDES

Average

Positive Composites
Number

9.9
6

9.7

5

8.7

3

17.9
6

10.2

5

Range

6.6-14.2

4.3-16.7

5.7-13.2

7.2-42.9

1.1-25.4

TOTAL BROMIDES

Average

9.0

0.54

3.5

4.0

3.2

Positive Composites

Number

6

4

2

5

2

Range

1.6-14.7

0.5-1.1

1.7-4.4

1.1-8.8

1.7-4.7

1 Six composite samples examined at each of the five sampling sites: Boston, Kansas City, Los Angeles. Baltimore, and Minneapolis.
Note; Bromide and arsenic values are reported on an "as is" basis for Dairy Products; Meat, Fish, and Poultry; and Oils, Fats, and Short-
ening.

TABLE 2b. — Pesticides found infrequently — by food class and region (June 1966 - April 1967)

[T = Trace <0.001 ppm]

Pesticide

No. Com-
posites

Amount
(PPM)

I (a). DAIRY PRODUCTS (8-13% fat) i
Residues in Parts Per Million — Fat Basis

ALDRIN

Baltimore

0.017

LINDANE

Kansas City

0.09

METHOXYCHLOR

Minneapolis

0.09

Kansas City

0,06

PGP

Boston

0.043

Kansas City

0.02

Baltimore

0.01

2,4, 5-T

Boston

0.19

HEPTACHLOR

Kansas City

0.02

ARSENIC

Kansas City

0.3

(AS2O3)

Boston

0.2

KELTHANE®

Los Angeles

0.03

2, 4-D

Minneapolis

0.027

II (a). MEAT, FISH, AND POULTRY (17-23% fat) :
Residues in Parts Per Million — Fat Basis

ALDRIN
PCP

Baltimore
Kansas City
Los Angeles

0.028

T

0.02

Vol. 1, No. 4, March 1968

Pesticide

District

No. Com-
posites

Amount
(PPM)

MEAT, FISH, AND POULTRY (17-23% fat) "—(Continued)
Residues in Parts Per Million — Fat Basis

LINDANE

Los Angeles
Kansas City
Boston
Minneapolis

0.03

0.012,0.374
0.014
0.010

PHORATE

Boston

O.OIO

2,4, 5-T

Boston

0.003

III (a). GRAIN AND CEREAL i
Residues in Parts Per Million

ALDRIN

Boston

0.01

BHC

Boston

0.015

CARBARYL

Kansas City

0.34

HEPTACHLOR
EPOXIDE

Los Angeles

0.001

PCP

Baltimore
Minneapolis

0.01
0.007

ARSENIC

Boston

0.10,0.12

(AS2O3)

Los Angeles

0.10

DDE

Boston
Kansas City

0.003
0.003

19

TABLE

2b.— PesliciJes found infrcq„nuh—hy food class and region (June 1966 - April /%7)— Continued

PsrnciDE

No. Com-
posites

Amount
(PPM)

III (a). GRAIN AND CEREAL '—(Continued)
Residues in Pans Per Million

TDE

Los Angeles
Kansas City
Los Angeles

1 10,001

1 10.012

2 i 0.001,0.004

IV (a). POTATOES'
Residues in Pans Per Million

CIPC

Kansas City

0.11

LINDANE

Boston

T

Los Angeles

0.002

TDE

Los Angeles

0.001,0.002.0.005

Boston

T

ARSENIC

Boston

0.10,0.14

( As.Oj)

Kansas City

0.2

PCP

Boston

T

DIELDRIN

Baltimore

0.006

Boston

0.006

Los Angeles

0.002

V (a). LEAFY VEGETABLES ^

Residues in Parts Per Million

BHC

Boston

T,T,0.002

Los Angeles

0.004

DIELDRIN

Kansas City

0.003

Boston

T

Los Angeles

0.001

DlTHIO=

CARBAMATES

Kansas City

0.8

Baltimore

0.44

ENDRIN

Los Angeles

0.05

LINDANE

Boston

0.007

Minneapolis

T

Kansas City

0.002.0.005

PARATHION

Boston

0.016

Kansas City

0.03,0.04

ENDOSULFAN

Los Angeles

0.003

CHIORDANE

Boston

0.02

ALDRIN

Boston

0.008

ARSENIC

Kansas City

0.4

(As-Oj)

METHYL

PARATHION

Kansas City

1

0.01

VI (a). LEGUME VEGETABLES ^
Residues in Parts Per Million

ALDRIN

Boston

1

T

LINDANE

Boston

1

T

ARSENIC

Boston

2

0.18,0.1

(AsKJj)

CHLORDANE

Boston

1

0.005

VII (a). ROOT VEGETABLES'
Residues in Parts Per Million

CARBARYL

ENDRIN

ARSENIC

(As:Os)
DITHIO=

CARBAMATES

VIII (a). GARDEN FRUITS'
Residues in Pans Per Million

ALDRIN

CARBARYL

DIAZINON

Boston
Kansas City
Boston

0.004,T

0.10

0.003

Pesticide

DISTKICT

No. Com-
posites

Amount
(PPM)

VII

(a). GARDEN FRUITS '—(Continued)
Residues in Parts Per Million

HEPTACHLOR

EPOXIDE
PARATHION

TCNB

IX (a). FRUITS'
Residues in Parts Per Million

CARBARYL

Kansas City

DIELDRIN

Boston

ETHION

Boston
Kansas City
Minneapolis

LINDANE

Kansas City

ARSENIC

Boston

(ASiOs)

Kansas City
Los Angeles

BHC

Los Angeles

METHOXYCHLOR

Boston

HEPTACHLOR

EPOXIDE

Boston

0.22

T

0.024

0.054

0.03

0.002

0.1

0.2

0.1

0.002

T

1

T

X (a). OILS, FATS. AND SHORTENING'
Residues in Parts Per Million

BHC

Boston

1

0.06

DIELDRIN

Kansas City

2

0.003.0.004

LINDANE

Los Angeles

0.007

Baltimore

0.004

Minneapolis

0.012

MALATHION

Boston

T

Kansas City

0.02

Minneapolis

0.052

Baltimore

0.062

PCP

Baltimore

0.02

ARSENIC

Boston

0.1

(AS:^0,,)

Los Angeles

0.1

ETHION

Boston

0.25

DIAZINON

Boston

T

XI (a). SUGARS AND ADJUNCTS'
Residues in Parts Per Million

ALDRIN

Baltimore

0.003

DDT

Los Angeles

0.002

Boston

T,T,

DDE

Boston

T

Minneapolis

T

Baltimore

0.02

LINDANE

Los Angeles

T

Boston

T,T

TDE

Minneapolis

0.02

ARSENIC

Boston

0.1.0.15,0.10

(Asi:03)

Los Angeles

0.1

PCP

Kansas City

0.01

MALATHION

Baltimore

0.07

KELTHANE®

Boston

1

0.015

XII (a). BEVERAGES'
Residues in Parts Per Million

LINDANE
PCP

ARSENIC
(AsiOa)
MALATHION

Boston
Boston
Boston

Baltimore

T

0.021
0.25

1 0.19

'Sin lomposiic samples examined at each of the five sampling sites; Boston. Kansas City. Los Angeles. Baltimore, and Minneapolis.

NOTE: Bromide and arsenic values are reported on an "as is" basis for Dairy Products; Meat. Fish, and Poultry; and Oils, Fats, and Shortenings.

20

Pesticides Monitoring Journal

RESIDUES IN FISH, WILDLIFE,
AND ESTUARIES

Pesticide Monitoring of the Aquatic Biota at the
Tule Lake National Wildlife Refuge^

Patrick J. Godsil and William C. Johnson

ABSTRACT

Because of pesticide poisoning of fish-ealinf; birds, the
Federal Water Pollution Control Administration established
a water quality monitoring program at the Tide Lake and
Lower Klamath Lake Wildlife Refuges in 1964. Over a
2-year period, samples of water, suspended material, sub-
merged aquatic plants, clams, and fish were collected and
analyzed for chlorinated hydrocarbon pesticides. Results are
reported from a typical station in the Tule Lake National
Wildlife Refuge.

Compounds DDE, DDD, DDT, chlordane, and endrin were
found regularly in samples of both water and biota. Water
contained a maximum of 0.100 ppb of endrin in 1965, while
tui chubs, Siphateles bicolor, accumulated a maximum of
198 ppb during the same year. Concentrations of endrin
in the other strata of the biota were distributed between
these extremes of the food chain.

The occurrence of endrin was directly associated with con-
taminated irrigation return water supplying the Refuge lakes.
As the concentrations of studied pesticides increased in the
drainage water, the biota also became contaminated. How-
ever, at the end of the .reason, as the concentrations de-
creased in the water, the biota was cleansed. Concentrations
in both water and biota returned to or near analytical
sensitivity (water — 0.007 ppb; biota — 4 ppb) between grow-
ing seasons.

Introduction

During the early 1960's, unusually large numbers of
fish-eating birds died at the Tule Lake and Lower
Klamath Lake National Wildlife Refuges (Fig.l).
Researchers of the U.S. Fish and Wildlife Service (5)
concluded that these deaths were caused by pesticide
poisoning resulting from the use of toxaphene and DDT

1 Klamath Basin Study. Federal Water Pollution Control Administra-
tion, Department of the Interior, 2261 South Sixth Street, Klamath
Falls, Oreg. 97601.

for pest control on agricultural lands within and sur-
rounding the Refuges. It was recognized that agricul-
tural drainage, carrying pesticides through the extensive
irrigation system supplying water to the Refuges,
presented a hazard to the wildlife. Consequently, at
the request of local groups and governmental agencies,
the Federal Water Pollution Control Administration
initiated the Klamath Basin Study. Its purpose was to
investigate water pollution problems associated with the
use of agricultural chemicals on lands of the Lost River
system draining into the Tule Lake and Lower Klamath
Lake Refuges. Specific objectives of the study were to:
(a) measure and identify pollutants responsible for the
wildfowl mortalities and, (b) determine the relation-
ships between land and water use and the pollutants.

An intensive monitoring program was initiated in April
1965 to obtain data on the occurrence of chlorinated
hydrocarbon pesticides at various locations within the
Lost River system. Monitoring stations were carefully
selected in and around the Wildlife Refuges. This paper
presents data showing the occurrence and fate of pes-

VoL. 1, No. 4, March 1968

21

ticiJc contaminants in various aquatic strata at a selected
reprcsontaiive station in the Tule Laice National Wild-
life Refuge.

Approximately 156.000 acres of land, irrigated prin-
cipally for the production of potatoes, grain, and pasture
grasses, lie upstream of the Wildlife Refuges (Fig. 1).
Irrigation supply water is used and reused through-
out this land before being discharged into the wetland
sumps of the Refuge. Such reuse creates a pollutant
buildup as water moves through the system. For pur-
poses of this paper, data obtained at drainage Pump B,
discharging into the Tule Lake sump, were chosen to
depict qualitative findings of the Project's pesticide
analyses. Pump B represents water that is used through-
out the irrigation system. Model studies indicate that this
water could he recycled on irrigated lands a maximum
of 5.2 times (<S). Consequently, this station quanti-
tatively represents water which contains high levels of
pesticides relative to levels found at other stations in
the study area.

Sampling Procedures

Monitoring stations were established at all significant
inflows and outflows to and from the Tule Lake and
Lower Klamath Lake Refuge areas. At Pump B, as
with the other stations, samples of water, suspended
material, submerged aquatic plants, clams, and fish were
collected for pesticide analysis.

Grab water samples were collected in two 1 -gallon glass
bottles. From each bottle, 1.5 liters were combined for
an analysis which was performed within 48 hours. Sus-
pended material (plankton, small vegetative fibers, sus-
pended solids) was collected by pumping 100 to 150
cubic feet of water through a 295-micron mesh net.
These samples, each weighing from 3 to 12 g, were
frozen while awaiting analysis. The aquatic plants (at-
tached algae and vascular) were obtained by raking or
handpicking. Pondweed (Pr>iamoi,'eion sp.) comprised
the majority of the vascular plants collected, but sig-
nificant amounts of watermilfoil (MyriophyUitm sp.)
and small amounts of other submerged aquatic plants
were also sampled. Cladophora sp. was the predominate
attached alga collected. These plants were frozen in
1-lb aliquots and stored for analysis. Native clams
(Gonklea sp.) ranging in length from 3 to 5 inches,
were collected using an Ekman dredge or a hand rake.
Five shucked clams were homogenized together and
frozen prior to analysis. Fish were collected using elec-
tric fishing gear or rotcnone. Approximately 909f of
these fish were tui chubs (Siphaleles hicolor) while the
others were blue chubs (Siphaleles i^ilii). All samples
of suspended materials, aquatic plants, clams, and fish
were wrapped in aluminum foil in the field to prevent
contamination.

In addition to sampling the natural biota, an in situ
study was made using largemouth bass (Microplents
salmoidex) and clams. These organisms were held in
separate submerged cages at the sampling station. Bass
were used because they can withstand confinement and
have been used extensively for pesticide bioassays. Tui
chubs were difficult to cage as they soon died from
disease. A baseline pesticide content was determined
before starting the //; silu study. Sufficient numbers of
both bass and clams were caged to allow sampling of
the exposed populations throughout the agricultural
season. Bass were held and sampled in this manner for
as long as 209 days.

All fish samples were prepared for analysis by homo-
genizing whole fish in a blender and then freezing them
for storage. Wild chub samples consisted of 5 to 20
individuals ranging in length from 2 to 7 inches, while
from 3 to 5 bass, 5 to 7 inches long, were sacrificed at
each increment of the cage study. No pathological ex-
aminations were made of their internal organs, as all
fish appeared to be in good health at the time of
sampling.

Laboratory A halysis

Chemical analyses for chlorinated hydrocarbon pesti-
cides provided individual^ results for DDE, DDT, DDD,
toxaphene, heptachlor, heptachlor epoxide, chlordane,
dieldrin, and endrin. However, the results of analyses
for these compounds in water and tissue samples showed
the compounds DDE, DDD, DDT, chlordane, and
endrin to be dominant. Other compounds either were
not present or were present in very small amounts —
i.e., <0.002 ppb in water and <4 ppb in ti.ssue — and
therefore were not included in this report
All analyses were conducted by the Klamath Basi-^ Study
laboratory utilizing gas chromatographic .e<-hniques in
conjunction with a microcoulometric titrating system
employing a silver cell for chlorinated hydrocarbon de-
tection. The cleanup procedures for sample extracts were
modifications of those presented by Mills (4). Following
cleanup, an equivalent of 1 .5 kg of water sample and
1 to 25 g of the other samples were injected into the
chromatograph. Identity of specific pesticides was con-
firmed by the use of several different columns. The
various columns used are as follows:

3% Dow-200 on acid washed 60-80 mesh Chrom-

osorb P, 'A" X 4-6'
Mixed column containing approx. equal parts of,

first, 5% FS-1265 and, second, 3% Dow-200 on

acid washed 60-80 mesh Chromosorb P, 'A" X

6'
3% OV-17 on 60-80 mesh Gas Chrom Q, 'A" X

6'
Sensitivity of the analytical results differs for each type
of sample analyzed and for each type of compound

22

Pesticides Monitoring Journal

detected. Also, varying quantities of material for a
specific sample and changes in instrument response add
to the variances of analytical sensitivity. The sensitivity
levels shown in Fig. 2 represent these variances. Table 1
shows present analytical sensitivities based on the in-
strument's normal operating capability.

Reproducibility of results was determined from statis
tical analysis of duplicate analyses of field samples.
Approximately 1 out of 40 samples was selected for
duplicate analysis. Table 2 shows the results for all
duplicate analyses as calculated by the following two
methods: (a) student's t-distribution for paired observa-
tions at the 95% confidence interval (3), and (b)
average percent deviation within laboratory (6). The
results for a particular sample and compound are based
on N samples with a mean value of X. Deviations from
the mean are expressed as a percent.

FIGURE 2. — Occurrence of endrin in water
and biota at pump B.

TABLE 2. — Reproducibility of analytical results

[£) C«f D I

TABLE 1. — Sensitivity of analytical results

Sample
Size

DDE, DDD,

Endrin
(PPB)

DDT,

Chlordane

(PPB)

Instrument

5 ng

15 ng

Water

1.5 kg

0.003

0.010

Fish

2.5 g

2.0

5.0

C Clams ^
J Vascular Plants 1
1 Filamentous Algae [
t Suspended Material J

10.0 g

0.5

1.5

Fish

Water

(CONC. <I00.0 PPB)

(CONC. <0.100 PPB)

z

z

o

o

»s

z

"1

z

N

X

N

X

2Q

uO

Ui >

> u

<D

t/) H

<a

DDE

23

31.4

14.2

36.3

4

0.015

53.4

26.6

DDD

13

26.2

15.3

39.9

—

—

—

—

DDT

15

29.4

27.2

36.9

—

—

—

—

Chlordane

21

28.5

48.3

34.2

—

—

—

—

Dieldrin

17

10.0

63.6

67.0

_

—

—

—

Endrin

18

13.8

62.8

56.5

6

0.031
0.049

25.8
34.7

25.8
30.6)

' Includes values >0.100 ppb.

No corrections of results were made for percent re-
coveries which averaged 80 and ranged from 71 to 95
for the reported compounds.

Results

Pesticides found at Pump B in the various aquatic
strata are directly related to the control of insects and
other pests plaguing the agricultural industry in and
around the Wildlife Refuge. These infestations require
the application of thousands of pounds of chemicals.
For example, during the 1966 growing season, ap-
proximately 14,000 lb of endrin were applied on the
study area at a rate of 1.6 Ib/acre/year. Portions of
these chemicals leach or fall directly into the drainage
canals and, as in the case of Pump B, are subsequently
discharged into the Wildlife Refuge.

Endrin has predominated in all analytical results, due
principally to its abundant usage and persistence in
drainage water after application. This, combined with
its acute toxicity (1.2) singles out endrin as the most
hazardous chlorinated hydrocarbon pesticide to wildlife
in the Klamath Basin.

The occurrence and fate of endrin in the aquatic strata
located in the Tule Lake sump at Pump B during 1965
and 1966 are shown in Fig. 2. Most significant is the
increase and subsequent decrease of endrin in all levels
of the biota during the main growing season (generally
from May through September). If a hydrograph of the
discharge from Pump B were shown, it would describe
a rise from zero in April to a peak in August, and a
fall again to zero in September. Correspondingly, the
various aquatic strata become contaminated, the con-
taminants increase to peak concentrations and fall to
near or below levels of sensitivity. Also emphasized, by
the generalized curves, is the relationship of the oc-
cun jnce of endrin in water to the subsequent contamina-
tion of the biota.

Vol. 1, No. 4, March 1968

23

For the 2 years shown, the level of peak contamination
is generally the same for both water and fish samples.
Water contained a maximum of 0.100 ppb in 1<)65 and
0.069 ppb in 1966, while captive fish accumulated a
ma.ximum of 97 ppb and 107 ppb of endrin, respectively.
Other strata of the biota vverc distributed between these
extremes of the food chain. Although a lesser number
of samples were taken during the off season, periodic
analyses revealed concentrations near or below the
laboratory's low sensitivity levels.
The days of exposure for caged bass and clams are
emphasized in Fig. 3 and 4. respectively. For the series
of bass and clams which were successfully maintained
over an entire irrigation season the rise and fall of
pesticide levels are again shown. These studies seem to
indicate that the accumulation of endrin is dependent
on the time of initial immersion. The greatest accumula-
tion for all series of bass and clams occurred beginning
in August of both 1965 and 1966. This rise reflects
increased endrin concentrations in water due to July
agricultural applications of endrin formulations.
Results of the analyses of water, aquatic biota, and
caged bass and clam samples taken at Pump B are
shown in Tables 3, 4, and 5, respectively. These tables
show results of analyses for other dominant chlorinated
hydrocarbon pesticides in addition to data for endrin.

FIGURE 3. — Diiys of exposure — caged boss

Conclusions and Discussion

Keeping in mind that the occurrence of chlorinated
hydrocarbons at Pump B is typical of similar drainage
flows in the Lost River system, we conclude that:

1. Agricultural practices in the Lost River system cause
chlorinated hydrocarbon pesticide contamination of
irrigation return water and the associated biota. This
fact, in addition to the reported effects of long-term
exposures of wildlife to low concentrations of
pesticides (7), indicates the hazards to wildlife, both
immediate and long-range, which must be considered.
These effects stress the need for water management
programs which eliminate, or at least alleviate, the
hazards from such contamination. Although no
wildlife mortalities due to pesticide poisoning have
been reported in the Klamath Basin Refuges in
recent years, this does not mean that detrimental
effects are not present. The continuing role of re-
searchers is to evaluate these effects in the lab-
oratory and eventually in the field. Until all the
answers are found, responsible water users and water
pollution control agencies must develop management
plans that minimize the occurrence of hazardous
materials discharged from agricultural lands.

FIGURE 4. — Days of exposure — caged clams

24

Pesticides Monitoring Journal

3.

Concentrations of pesticides in water and biota of
the Lost River system increase to peaiv values during
the summer growing season, then decrease to near
or below levels of laboratory sensitivity after the
close of the season.

Most important, this fact demonstrates that short-
term pesticide contamination of an aquatic environ-
ment does not establish permanent residual con-
centrations of pesticides in the various strata. The
dilution effect of irrigation water applications after
final pesticide treatment is certainly the cause of this
beneficial cleansing. Consequently, flushing with
uncontaminated water is a means of controlling
pesticide levels in the natural food chain of fish-
eating wildfowl.

From year to year the concentration of studied
pesticides in the aquatic strata of the Lost River
system was no greater than the previous year's peak
level.

The above conclusions bring out the point that, in
agricultural areas with only a short summer growing
season, the levels of contamination in the aquatic
environment are governed by the seasonal variations,
thereby limiting accumulations to the seasonal peaks.
For the agricultural community within the Lost
River Basin, this fact tempers one of their primary
anxieties concerning pesticide usage. At the same
time that the wildfowl mortalities were occurring
in the Refuge, considerable national attention was
focused on the effects of residual pesticides on the
Nation's wildlife. Naturally, the first reaction was
one of concern that runoff from the irrigated lands
in the basin might be causing a continuing buildup
of pesticide concentrations in the National Wildlife
Refuges which would result in continued mortalities
in the migratory bird population. Under present
land practices, this continued accumulation is not
occurring in the aquatic biota of the basin.

TABLE 3. — Pumh B water analyses
[ — = Results below analytical sensitivity]

Residues—

-PPB

Date

Collected

DDE

DDD/DDT 1

Chlordane

Endrin

4/01/65

0.006

5/14/65

—

—

—

—

6/22/65

0.005

—

—

—

7/22/65

0.010

—

—

—

8/11/65

0.010

—

—

0.100

8/27/65

0.003

—

—

0.015

9/10/65

0.007

—

—

0.017

10/01/65

0.003

—

—

0.007

10/11/65

0.007

—

0.010

—

10/20/65

0.003

—

—

—

11/09/65

0.010

0.010

0.013

—

1/18/66

0.003

—

0.013

—

2/17/66

0.003

—

—

—

3/22/66

0.003

—

0.010

—

4/26/66

—

—

0.010

—

6/07/66

0.027

0.027/0.027

0.100

—

6/22/66

—

—

0.013

—

7/06/66

0.003

—

0.010

—

7/20/66

0.003

—

0.012

—

7/28/66

—

—

— :

—

8/04/66

—

—

0.013

—

8/05/66

—

—

—

—

8/09/66

—

—

—

0.008

8/11/66

—

—

—

—

8/18/66

—

—

—

0.023

8/20/66

—

—

—

0,011

8/22/66

—

—

—

0.021

8/24/66

—

—

—

0.069

8/26/66

—

—

—

0..057

8/29/66

—

—

—

0.056

9/02/66

—

—

—

0.030

9/07/66

—

—

—

0.007

9/13/66

—

—

—

0.010

9/22/66

—

—

—

0.007

9/29/66

—

—

—

0.010

10/04/66

0.003

—

—

—

10/19/66

0.006

0.002/0.013

—

0.009

11/04/66

0.004

0.017

0.051

—

11/18/66

0.003

—

—

—

11/25/66

0.003

0.007

0.017

0.007

11/30/66

—

—

—

—

12/14/66

—

0.017

—

—

1/11/67

—

—

—

—

2/06/67

0.003

0 /O.OIO

0.017

—

' Single values represent a total response, i.e., where DDD and DDT
could not be separated.

Vol. 1, No. 4, March 1968

TABLE 4. — Chloriinilcd hydrocarbon pesticides contained
in the biota at Pump B

[ — = Results below analytical sensitivity]

Date
Collected

8/10/65
12/28/65
7/22/66

Residues— PPB

DDE

DDD/DDT 1

Chlordane Endrin

Suspended Material

4/20/66

_

0.75

3.0

1.5

6/22/66

—

—

67.0

6.0

7/22/66

1.7

10.0

6.0

1.3

8/22/66

6.6

4.0

6.0

57.7

9/13/66

—

4.0

8.0

13.0

10/26/66

1.0

0.7/2.0

1.5

5.3

11/16/66

—

—

8.5

—

1/06/67

1.5

3.3/12.0

14.7

1.5

Vascular Plants

6/22/66

1.0

1.0

5.0

. —

7/22/66

—

—

2.0

1.6

8/22/66

1.0

10

2.0

12.2

9/13/66

—

—

1.5

12.5

9/29/66

0.8

1.2

—

4.8

10/20/66

1.0

10.0

6.0

8.0

11/16/66

0.6

0.7

2.6

1.8

Algae

4/20/66

0.5

0.75

2.0

2.0

6/22/66

2.0

3.0

50.0

—

7/22/66

—

—

—

—

8/22/66

0.8

0.4

1.7

22.3

9/13/66

1.3

1.3

13.5

10.8

Chubs

8/27/65

45.0

17.0

198.0

4/20/66

26.0

12.0

24.0

10.0

6/22/66

14.0

10.0

10.0

6.0

7/22/66

6.2

9.6

8.0

4.0

8/22/66

2.5

2.5

—

30.5

Clams

4.0
4.0
4.8

4.0
3.0
4.8

3.0
4.5
12.0

34.0
4.0
2.0

' Single values represent a total response, i.e.
could not be separated.

where DDD and DDT

25

TABLE 5. — Chlorinated hydrocarbon pesticides in
targemoiith hass and clams held in cages at Pump B

1— =r Results below

analytics

1 sensitivity]

Residues — PPB

Date
Collected

Days
Ex-
posed

Number
Col-
lected

DDE

DDD/
DDT'

Chlor-

DANE

Endrin

Largemouth Bass

GROUP 1

7/01/65

0

5

27

10

—

—

7/16/65

15

5

32

18

—

2

7/30/65

29

5

37

14

—

31.5

9/24/65

85

2

38

23

—

74

11/05/65

127

1

19

16

15

65

GROUP 11

8/17/65

0

5

27

6

—

2.5

8/18/65

1

5

25

50

—

20.2

11/05/65

80

1

22

13

13

97

11/19/65

94

3

26

15

20

72

GROUP 111

10/07/65

0

5

34

19

11

6.5

11/05/65

29

5

38

14

17

20

11/19/65

43

2

14

17

43

19

GROUP IV

6/15/66

0

5

16

16

8

—

7/14/66

29

5

37.5

31

33

2

8/16/66

62

4

11

11

—

20

9/16/66

93

5

12.5

9.5/6.5

—

107

9/30/66

107

5

12.5

10/10

7.5

40

10/13/66

120

5

12

10/8.8

8

65

1/10/67

209

2

21.2

8.9/7.3

—

15.3

Clams

GROUP I

12/14/65

0

5

2.5

4

25

12/28/65

14

4

2

2

8

1.7

1/28/66

45

3

2.5

5

5

3.5

3/04/66

80

3

2.25

2

6

—

3/30/66

106

2

1

1

—

2.5

GROUP II

3/30/66

0

5

1

—

—

—

5/02/66

33

5

1.5

3

4

—

6/22/66

84

5

2

2

4

2

7/14/66

106

5

2

4

3

—

8/16/66

139

4

—

—

2

2.5

8/30/66

153

5

0.75

—

—

90

9/16/66

170

5

—

—

3

18

9/30/66

184

5

—

—

6

14

10/13/66

197

5

3

2

—

12.6

11/16/66

231

5

6.3

2.0/3.0

3

7

1 10 67

255

5

2.4

2.3

5.1

5.9

The chemical names of compounds mentioned in this paper are:
I. l-dichloro-2,2-bi5(r-chlorophenyl) ethylene
1.1.1 -t r ichloro-2,2-bis( p-chlorophenyl ) ethane
l.l-dichloro-2,2-bis(p-chlorophenyl)ethane

DDE
DDT

DDD

Chlordane

' Single values represent a total response, i.e., where DDD and DDT
could not be separated.

1. 2.4,. "i .6,7,8. 8-octachloro-3a,4.7,7a-tetrahydro-
4.7-methanoindane

Endrin l.2.3,4.IO,U>-hexachloro-6,7-epoxy-l,4,4a,5,6.7,8,8a-

octahydro-l,4-cMdo-fnJo-5,8-dimethanonaphthalene

Heptachlor 1 ,4. 5,6.7,8. 8-heptachloro-3a,4,7,7a-tetrahydro-

4.7-methanoindene

Heptachlor epoxide 1.4. 5.6.7.8. 8-heptachloro-2.3-epoxy-3a,4,7,7a-
lelrahydro^.7-methanoindan

Dicldrin not less than 85^r of 1.2.3.4,10,10-hexachloro-6,

7-epoxy- 1 ,4,4a. 5,6, 7,8. 8a-octahydro-l .^-endo-exo-
5,8-dimethanonaphthalene

Toxaphene chlorinated camphene containing 67% to 69%

chlorine

Acknowledgments

The authors wish to acknowlecJge Messrs. Robert E.
White and Gerald L. Muth who were, successively, in
charge of laboratory operations and are responsible
for the analytical results presented. We also wish to
thank the many staff members of the Klamath Basin
Study for their efforts in providing the reported subject
material.

LITERATURE CITED

(/) Henderson. C. Q. H. Pickering, and C. M. Tarznell.
1959. Relative toxicity of ten chlorinated hydrocarbon
insecticides to four species of fish. Trans. Amer. Fish
Soc. 88:23-32.

(2) Katz. M. 1961. Acute toxicity of some organic in-
secticides to three species of salmonoids and to the
threespine stickleback. Trans. Amer. Fish Soc. 90:264-
268.

(.?) Li. J. C. R. 1957. Introduction to statistical inference.
Edwards Brothers, Inc. Ann Arbor, Mich.

(4) Mills, P. E. 1959. Detection and semiquantitative esti-
mation of chlorinated organic pesticides in food by
paper chromatography. J. Ass. of Agr. Chem. 42:734.

(5) Pillmore, R. E. 1961. Pesticide investigations of the
1960 mortality of fish-eating birds on Klamath Basin
wildlife refuges. U. S. Fish Wildlife Serv., Wildlife
Res. Lab. 12 p.

(6) Robert A. Taft Sanitary Engineering Center. 1966.
Water Pesticides No. 1. Study Number 24. Public
Health Serv. Publication No. 999-WP-39. 64 p.

(7) Riidd, R. L. 1964. Pesticides and the living landscape.
Univ. of Wis. Press, Madison, Wis. 320 p.

(5) Woods, Philip C. and Gerald T. Orlob. 1963. The Lost
River system — a water quality management investiga-
tion. Water Resources Center Conlrib. No. 68, Univ. of
Calif., Berkeley, Calif. 54 p.

26

Pesticides Monitoring Journal

Chlorinated Pesticide Residues in an Aquatic
Environment Located Adjacent to a Commercial Orchard

R. J. Moubry', J, M. Helm-, and G. R. Myrdal'

ABSTRACT

Samples of water, silt, bottom organic debris, bottom
organisms, and fish were collected from an aquatic environ-
ment located adjacent to a commercial orchard. Residue data
obtained from the analysis of these samples are presented.
The results obtained indicate that contamination of the
environment studied was minimal.

Introduction

Pesticides, principally the chlorinated hydrocarbons,
have been used extensively in Wisconsin orchards in the
production of fruit for market. In 1966, an exploratory
investigation was conducted by the Wisconsin Depart-
ment of Natural Resources to evaluate the effects of
such pesticide usage on the aquatic environment of
streams located in the drainage area of these orchards.

Knights Creek, located in Dunn County, Wis., was
selected as the site of this investigation. The upstream
area of this creek branches to the north and to the
south. A commercial orchard is located on top of a
hill at the confluence of these two branches traversing
along the base of the hill. Accurate records of pesticide
usage were unavailable, but it was ascertained that 150
acres of the orchard had been treated with endrin for
rodent control at a rate of approximately 1 lb/ acre
actual in the fall of 1963, 1964, and 1965. During this
same 3-year period, approximately 100 lb actual of
dieldrin also had been used each year in foliar treatment
of the entire orchard (195 acres), and, during the
period 1955 to 1962, approximately 50 lb actual of
dieldrin had been applied yearly to this orchard. Many
other types of pesticides, including DDT, also had been
used in this orchard, but the total amounts applied were
not determined.

^ Wisconsin Department of Agriculture, General Laboratory Division,
Bureau of Ciiemistry, 4702 University Ave., Madison, Wis. 53702.

- Wisconsin Department of Natural Resources. Division of Resource
Development. Bureau of Water Resources, 421 State Office Building,
Madison, Wis. 53702.

Sampling Methods

Sampling stations were established in the north and
south branches of Knights Creek, at the confluence of
the two branches, and in a control area located in a
tributary of the north branch. On March 8, 1966,
samples of silt, bottom organic debris, and bottom
organisms were taken at each sampling station with the
aid of a dredge which collected stream bottom material
to a depth of 3 to 4 inches. The organisms and organic
debris were then removed from the material, and 1 -quart
portions each of the separated organic matter and
remaining silt from each of the sampling stations were
taken for analysis. Bottom organisms were first separated
by species; however, in some instances, difficulty in
obtaining a sufficient quantity of individual species
necessitated the compositing of different organism
species into a single sample. Bottom organism samples
were then held in a formaldehyde solution.

A 5-quart sample of runoff ground water entering the
stream was collected at each of the sampling station
areas on June 1, 1966, either during or immediately
after a heavy rain storm. Due to a heavy turf surround-
ing this stream, these water samples were to all appear-
ances devoid of silt.

The fish samples were collected on August 24, 1966, by
means of an electro-fishing apparatus. Fish were un-
available in the control area at the time of sampling.

Analytical Methods

The samples of silt were air-dried to approximately 15%
moisture and sieved. The material which did not pass
through a No. 8 sieve was discarded. The sieved silt
samples were extracted by the hexane-acetone procedure
(/), and the silt extracts were then cleaned up with
Florisil (2). The debris samples were ground, mixed,
and extracted by the acetonitrile-water extraction pro-

i

Vol. 1, No. 4, March 1968

27

ccdure (.'). A portion of each homogenous sample of
silt and debris was taken for moisture determination.
Analysis was made on the "as is" basis. The dry weight
residue resuhs were oht;iined by calculation, using the
percent moisture obtained from each sample.

The bottom organism samples, submitted in formalde-
hyde solutions, were drained. Each of the formaldehyde
solutions was then analyzed for chlorinated hydrocarbon
pesticide residues and interfercnt gas chromatographic
peaks. None were found. Some of the bottom organisms
(caddis fly larvae) were incased in a sand covering.
These were removed and discarded prior to grinding.
The drained and decased organisms were ground, ex-
tracted, and cleaned up (2). The sample size used for
analysis ranged from 8 to 10 g. The results obtained
were reported on the drained weight basis.

The fish collected from each sampling station were
pooled by species. The number of fish composited into
each sample is shown in Table 3. The fish samples were
ground as received. The ground samples included head,
tail, scales, and viscera. The samples were extracted and
cleaned up (2), with results being reported on the ex-
tracted fat basis. The percentage of fat in the samples
was determined and reported.

The water samples (4.800 ml each) were extracted
three times with redistilled hexane. The extracts were
concentrated and cleaned up with Florisil.

Determination of the amount of pesticide residues pres-
ent in the samples was by electron capture gas-liquid
chromatography. The instrument used was a Jarrell-Ash,
Model 28-710, gas chromatograph. The column packing
systems used were 10% DC-200 on Anakrom ABS, and
a mixed bed column consisting of nine parts 10% DC-
200 and five parts lO'^'r QFI on Gas Chrom Q.

The sample size, final volume of sample extract, and
amount injected into GLC were adjusted to provide a
sensitivity of 0.001 ppm dieldrin for the silt, organic
material, bottom organisms, and fish tissue. The level
of detection for the water samples was 25 ppt of dieldrin.
Inasmuch as this was an exploratory survey, recovery
studies were not conducted in conjunction with analysis
of these samples. Recovery studies are run at periodic
intervals in the laboratory to insure reliable analysis
and are in the range of 90% . Due to the minimal amount
of residue detected in the majority of these samples,
confirmation of the residue detected was restricted to
multiple GLC column technique. The data presented are
the results obtained using the methodology specified.

Discussion

The results obtained are presented in Tables 1 through 4.
No residues were detected in the orchard runolT water
entering the stream on the date these samples were

collected. No detectable DDT or its analogues were
present in the silt and debris samples. Low levels of
DDT and its analogues were detected in the bottom
organisms. The DDT and dieldrin residues detected in
the brook trout were at the same general level as those
detected in the same and similar species collected and
analyzed in a recent State-wide residue-in-fish survey
(4). Although low-level cndrin residues were detected
in the silt, organic matter, and bottom organisms, none
were detected in the fish samples. Evaluation of the re-
sults obtained in this limited investigation indicates that
the pesticide usage in the orchard has not significantly
contaminated the aquatic environment of this adjacent
creek.

Tlie chemical names of compounds mentioned in this paper are:
Dieldrin

Endrin

DDT
DDD
DDE

not less th.in 85Tr of l.2.3.4.10.I0-hexachloro-6.7-epoxy-
l,4.4a.5,6.7,8,8a-octahydro-1.4-eM(/o-exo-5.8-dimethano=
naphthalene

1.2.3.4.IO,in-hexachloro-6.7-epoxy-1.4.4a.5,6,7.8.8a-
octahydro-1.4-f/i(/f>-e/i(/o-5.8-dimethanonaphthaiene
l.l.l-trichloro-2.2-bis(p-chloropheny Methane
I.I-dichIoro-2,2-bis{p-chlorophenynethane
l,l-dichloro-2,2-bist p-chlorophenyl) ethylene

Acknowledgments

The authors wish to acknowledge the assistance of R.
Smith (Warden) and L. Frankenburger (Biologist) of
the Division of Conservation, who assisted in the col-
lection of samples, and L. Lueschow of the Division of
Resource Development, who performed the identification
of invertebrate organisms.

LITERATURE CITED

(/) Lichtcnslcin. E. P.. G. R. Myrdal, and K. R. Schulz.
1964. Effect of formulation and mode of application of
aldrin on the loss of aldrin and its epoxide from soils and
their translocation into carrots. J. Econ. Entomol.
57:133-136.

(2) Barry, Helen C, Joyce G. Hundley, and Loren Y.
Johnson. Pesticide Analytical Manual Vol. 1, 2.21 (A),
U. S. Department of Health, Education, and Welfare,
Food and Drug Administration, Washington, D. C.
20204.

(.*) Ihid.. 2.21(B).

(4) Kleinerl, S. J., P. E. Degur.^e, T. L. Wirlb, and L. C.
Hull. 1967. DDT and dieldrin residues found in Wis-
consin fishes from the survey of 1966. Preliminary
Report, Research Report No. 23 (Fisheries), Wisconsin
Conservation Department, Research and Planning Divi-
sion, Madison, Wis.

TABLE 1. — Chlorinated hydrocarbon pesticide residues
detected in silt and debris samples

[ — = None detected]

Residues in PPM— Dry Weight Basis

Srrs

Silt Samples

Debris Samples

Endrin

Dieldrin

Endrin

Dieldrin

Control
North Branch
South Branch
Confluence

0.003
0.002
0.013

0.005

0.025
0.011
0.014

0.004
0.006
0.002
0.002

28

Pesticides Monitoring Journal

TABLE 2. — Chloriiwlcd hyilrncarhon pesticide residues delected in bottom or^anisiyt samples

Sample

Residues in PPM — Whole Weight Basis

Site

DDE

DDD

DDT

DDT AND

Analogues

Dieldrin

Endrin

Control

Caddis Fly Larvae
(Limnephilus rhombicus)

Organism Composite ^

0.014
0.013

0.009
0.008

0.010
0.012

0.033
0.033

0.002
0.001

—

North Branch

Alder Fly Larvae
(.Sialis sp.)

Fresh-Water Shrimp
(Gammarus sp.)

Caddis Fly Larvae
{Limnephilus rliombicus)

0.005
0.010
0.006

0.003
0.007

0.007

0.008
0.012
0.011

0.016
0.029

0.024

0.013
0.003

0.002

0.009
0.025
0.003

South Branch

Organism Composite =

0.007

0.011

0.016

0.034

0.002

0.004

Confluence

Fresh-Water Shrimp
{Gammarus sp.)

0.009

0.007

0.015

0.031

0.013

0.013

' Consisted of Gammarus, Agapelus. Protoptila, caddis pupae, Dytiscidae. Atherix variegala. immalure stone flies. Procladius, Hydrobaeninae,

and Calopsectra.
-Consisted of Gammarus, Sialis. Isoperla bilineata, Protoptila, caddis pupae, Agapetus. Cheumatopsyche. aquatic earthworm, Tipula, Tabanidae,

Procladius, immature Coleoptera. Hydrobaeninae, Limnephilus rhombicus, and Potomyia.

TABLE 3. — Chloritiated hydrocarbon pesticide residues detected in fish samples

Sample

No. OF
Fish

Per-

Residues in PPM

Site

Fat Basis

Whole Weight Basis

Fat

DDT AND

Ana-

DDT AND

Ana-

DDE

DDD

DDT

logues

Dieldrin

logues

Dieldrin

North Branch

Brook Trout
(Salrelinus fontinatis)

2

4.0

1.41

1.04

1.42

3.87

0.26

0.155

0.014

Northern Creek Chubs
(Semotilus atromaculatus)

24

3.8

1.02

0.67

0.12

1.81

0.34

0.069

0.013

Muddlers
(Cottus bairdi)

17

2.4

0.65

0.45

1.47

2.58

0.69

0.062

0.017

South Branch

Brook Trout
{Salvelinus fontinalis)

4

4.6

0.83

0.56

1.15

2.54

0.18

0.168

0.008

Northern Creek Chubs
(Semotilus atromaculatus)

33

3.6

1.00

0.53

0.59

2.12

0.17

0.076

0.006

Muddlers
(Coitus bairdi)

16

2.4

0.57

0.47

1.54

2.58

0.31

0.062

0.007

Confluence

Brook Trout
(Salvelinus fontinatis)

1

4.6

0.29

0.26

0.37

0.92

0.21

0.042

0.010

Northern Creek Chubs
(Semotilus atromaculatus)

6

2.6

1.53

0.63

0.20

2.36

0.31

0.061

0.008

Black Nosed Dace
(Rhinichtliys atralutus)

10

6.0

1.92

0.78

0.10

2.80

None

0.168

None

Muddlers
(Cottus bairdi)

4

2.2

0.55

0.40

0.54

1.53

0.41

0.034

0.009

Note: No endrin residues were detected in these samples.

TABLE 4. — Results of analyses of rain runoff water for
chlorinated hydrocarbon pesticide residues

Site

Ml H-O Extracted

Pesticide Residues '

Control

4800

None detected

North Branch

4800

Do.

South Branch

4800

Do.

Confluence

4800

Do.

1 Minimum level of detection was 25 ppt of dieldrin.

Vol. 1, No. 4, March 1968

29

PESTICIDES IN SOIL

Monitoring the Effects of the 1963-64 Japanese Beetle Control Program on Soil, Water,

and Silt in the Battle Creek Area of Michigan

J. E. Fahey', J. W. Butcher, and M. E. Turner'

ABSTRACT

The 1963-64 Japanese beetle cnnlrol program in Buttle
Creek, Mich., ivas monitored hy Michigan Stale University
and tlie Agricultural Research Service. U. S. Department of
Agriculture. Soil, water, and sill samples were obtained after
treatment of infested areas with 20 lb of 107c granular
dicldrin per acre. Dieldrin was present in only .? of 22
pre-treatmcnt soil samples. It averaged 1.25 ppm in soil
samples collected on November 23. 1963, just after treat-
ment, and 1.39 ppm on June 25, 1964. No detectable resi-
dues of dieldrin were present in water after treatment, and
residues in silt were low, absent, or inconclusive due to
interferences.

Ittiroduction

The 1963-64 Japanese beetle (Popillia japonica New-
man) control program in the Battle Creek area of
Michigan was cooperatively undertaken by the Ento-
mology Research Division and the Plant Pest Control
Division. Agricultural Research Service, U.S. Depart-
ment of Agriculture; the Plant Industry Division, Mich-
igan Department of Agriculture; and the Entomology
Department. Michigan State University. The area
treated consists of 12.601 acres. Dieldrin was used at the
rate of 2 lb technical per acre {\07( dieldrin at 20 lb
granular per acre ) . The objectives of the program were
to treat the city of Battle Creek and surrounding sub-
urban area and kill as many beetles as possible. The
program started on October 27, 1963, and ended April
15, 1964. No operations took place from December 14,
1963. to March 30. 1964. The applications were made
with ground equipment, including two buffalo turbines
and two Skibee spreaders mounted on pickup trucks,
and hand-operated Seymour seedcasters.

' Enlomology Ucpanmcnt. Purdue University, formerly in Charge
Analytical InvestiKations, Pesticide Chemicals Research Branch. En-
tomolouy Research Division. Agricultural Research Service. U. S.
Department of Atfricullure.

= Department iif Entomolony. Michigan Stale University.

' Plant Pest Control Division. Agricultural Research Service. U. S.
Department of Agriculture.

Precautions were taken wherever possible to prevent
contamination and hazardous residues. Special care was
taken to avoid getting dieldrin into lakes, rivers, and
creeks. Only small sections of shoreline were treated
between rains. Great care was taken also to keep the
insecticide off sidewalks, streets, driveways, etc. Feeding
dishes for pets, sand boxes, and bird baths were turned
over or covered with sections of tarpaulin before
treatment. Several pastures, small hayfields, and garden
areas with sensitive crops were bypassed in compliance
with label recommendations for dieldrin.

The monitoring program was conducted by the Ento-
mology Department, Michigan State University; and the
Pesticide Chemicals Research Branch. Entomology Re-
seach Division. Agricultural Research Service. The work
by Michigan State University was supported by con-
tracts with the Plant Pest Control Division, AP^.

All collections were made by or under the direction of
Dr. J. W. Butcher, and all residue analyses were per-
formed by or under the direction of Jack E. Fahey.

A preliminary survey of the occurrence and distribution
of chlorinated hydrocarbon insecticide residues in soil
from Battle Creek. Mich., was reported by Fahey,
Butcher, and Murphy in 1965 (/). They found dieldrin
in only 17 of 227 samples. The dicldrin residues found
rangcti from 0.06 to 2.2 ppm. Only one sample con-
tained more than 1() ppm of dicldrin.

Collection of Samples

Prior to the start of control operations, twenty 1- by
3-inch soil cores were collected from sod and twenty
1- by 3-inch cores from garden or shrub-planted (culti-
vated) areas in one city lot per 40 acres. The lot chosen
for sampling was always on the extreme southwest
corner of each 40 acres. If, for any reason, the sample

30

Pesticides Monitoring Journal

could not be obtained at the preselected point, the col-
lector sampled the closest accessible lot.
The sod and cultivated soil samples were packaged and
analyzed separately.

Similar soil samples were obtained at every tenth point
as soon as possible after treatment — in November 1963
— and again in June 1964.

Water samples were collected from 1 3 points in ponds,
creeks, and the Kalamazoo River. Collection points were
established throughout the entire treatment area in order
to detect residues that might be washed off the treated
soil surface into the major drainage pathways. A 1 -gallon
sample of water was collected from preselected points
before treatment on October 31 and November 2, and
after treatment on December 19, 1963, and March 23,
1964.

Silt samples were collected from the streams and ponds
at the water collection points on October 3 1 and Novem-
ber 2 (pre-treatment) and on December 19, 1963 (after
treatment). Collections were discontinued after the first
post-treatment sample.

Preparation of Samples for Residue A nalysis

SOIL AND SILT

Recovery of Residue: Silt samples were filtered, dried,
and ground before analysis. Soil samples were sieved and
dried. Aliquots were weighed and 10% moisture added.
The samples were then stripped with a 2:1 mixture of
he.xane and isopropyl alcohol, using 2 ml per gram of
soil (or silt). The alcohol was removed by washing with
water; the hexane was dried over sodium sulfate.
Cleanup: A 40-ml aliquot was reduced to 10 ml and
chromatographed on a 4:1 magnesia celite column (as
used in colorimetric analysis).

Analysis: Suitable aliquots of the cleaned residue solu-
tion were injected into a Jarrell-Ash gas-liquid chro-
matograph, electron capture detector.

Critical Temperatures:

Oven 175 C

Injector 235 C

Splitter 210 C

Detector 200 C

Column: '4" X 4' aluminum

2% SE 30 on Anakrom ABS

Results of analyses were qualitatively verified by thin
layer chromatography.

WATER

Residue Recovery: Water samples of approximately 2
liters were extracted with 200 ml normal hexane for 5
minutes. The hexane extract was dried over sodium
sulfate.

Analysis: Analyses were made by gas-liquid chro-
matography, using the same instrument and conditions as
for soil analyses.

Results of the A nalysis

Table 1 lists dieldrin residues recovered from pre- and
post-treatment soil samples collected at Battle Creek.
The number of dieldrin granules visible in four 1 -square-
foot soil surface counts per collection point are given
along with ppm dieldrin residues recovered from the
same points before and after treatment. Table 1 also
shows other chlorinated hydrocarbon residues found in
pre-treatment samples.

Table 2 shows the results of analysis of pre- and post-
treatment water and silt samples. There were no verifi-
able residues detected in any of the post-treatment water
samples. Because of the low residues found in post-
treatment silt samples and interferences in analysis, the
silt sampling was discontinued after one sampling.

Summary and Conclusions

Analysis of soil, water, and silt samples from the Battle
Creek treatment area was carried out. The findings may
be summarized as follows:

Substantial, although not uniform, residues of hepta-
chlor, chlordane, BHC. DDE, or p.p'-o.p'-DDT were
present in virtually all samples taken from turf and
cultivated plots throughout the city of Battle Creek
before treatment. Only three pre-treatment samples con-
tained measurable dieldrin residues.
Counts of dieldrin granule distribution and levels of
dieldrin residues in soil after treatment showed that
coverage was almost complete and probably adequate
for control. The soil samples collected on November 23,
1963, contained an average of 1.25 ppm of dieldrin while
those collected June 25, 1964, contained an average of
1.39 ppm of dieldrin.

No detectable residues were present in water on the dates
sampled after treatment.

Dieldrin residues in streambed or pond silt were low,
absent, or inconclusive due to interferences.

Michigan Agricultural Experiment Station Publication
No. 4267

A cknowledgment

Grateful acknowledgment is made to Mr. Calvin Corley,
U.S. Department of Agriculture, Entomology Research
Division, Pesticide Chemicals Research Branch. Belts-
ville, Md., for assistance in the analytical work.

LITERATURE CITED

(/) Faltcy. J. E.. J. W. Biilclur. ami R. T. Miiipiiy. 1965.
Chlorinated hydrocarbon insecticide residues in soil of
urban areas. Battle Creek, Mich. J. Econ. Entomol.
58:1026-27.

Information in this paper is not for publication without prior approval
or for use in sales promotion or advertising which expresses or implies
endorsement of the product by the U. S. Department of Agriculture.

Vol. 1, No. 4, March 1968

31

TABLE 1. — Residues of chlorinalcJ hydrocarbon in.icclicidcx in soil samples from Bailie Creek, Mich.

Type'

DlFl-DRIN

Granule
Count"

Pesticide Residues in Soil Samples

Site

Pre-Treatment

Post-Treatment
Dieldrin

BHC

DDE

o.p'-DDT

p.p'-DDT

Chlordane

Hepta-
chlor s

Dieldrin

11/23/63

6/25/64

1

T

10-17-15-18

.03

.11

.20

»0.84

3.30

2

C

—

.03

—

.17

.10

—

*0.6I

0.15

3

T

6-14-0-2

—

.03

—

1.60

.10

—

0.35

1.35

4

C

—

—

—

.21

—

—

—

0.58

0.34

5

T

21-8-12-6

—

—

—

—

—

—

—

3.06

3.00

6

C

—

—

—

.07

—

—

—

0.73

4.40

7

T

3-2-3-0

—

.31

.04

.51

—

—

—

0.86

1.50

8

C

—

.05

.04

.22

—

—

—

1.84

1.20

9

T

19-18-5-10

.20

.20

.30

1.30

—

_

—

8.63

3.10

10

T

5-2-3-3

—

—

—

.15

.10

—

0.1

0.28

3.00

11

C

—

—

—

—

—

—

—

2.33

3.00

12

T

0-0-0-0

.10

—

—

.80

120.0

1.6

—

0.06

0.13

13

C

—

.14

.20

2.10

0.5

—

—

0.37

0.13

14

T

3-18-11-10

—

—

—

.07

0.13

—

—

1.01

1.00

15

C

—

—

—

.08

—

—

—

2.21

2.70

16

T

2-3-1-2

—

_

—

—

—

—

—

0.03

0.26

17

T

12-8-6-26

—

—

—

.07

—

—

—

0.65

0.30

18

C

—

.03

.04

.30

—

—

—

—

—

19

T

21-7-2-3

—

—

—

.08

.10

—

—

1.26

0.14

20

C

—

—

—

.07

.13

—

—

0.58

0.15

21

T

3-3-1-7

.02

—

—

—

—

—

0.07

0.04

0.01

22

C

—

—

.25

—

—

0.07

0.05

O.OS

» T = Turf sample: C = Cultivated soil.
'Granules found in a unit of space treated.
' Includes hept.ichlor epoxide.
•Sampled 11/ 11 /S.'t instead of 11/23/63.

The chemical names of compounds mentioned in this paper are:

Dio'drin not less than 85% of l.2,3.4.10.10-hexachloro-6,7-epoxy-l,4,4.i.5.6.7,8.8a-octahydro-1.4-endo-Mo-5,8-dimethanonaphthalene

DDE l.l-dichloro-2.2-bis(p-chlorophenyl )ethylene

BHC l,2,.V4.5,6-hexachlorocyclohexane, mixed isomers

Chlordane 1. 2.4.5,6,7.8. 8-octachloro-3a.4,7,7a-lelrahydro^,7-methanoindane

o.p'-DDT. p.p'-DDT I, l,l-Irichloro-2,2-bis(p-chlorophenyl) ethane

Heptachlor 1,4,5.6.7, 8, 8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methanoindene

Hcptachlor epoxide 1 ,4,5,6,7,8,8-heptachloro-2,3-epoxy-3a,4,7,7a-tetrahydro-4.7-melhanoindan

32

Pesticides Monitoring Journal

TABLE 2. — Dieldrin residues in water and sill from ponds and streams in Battle Creek, Mich.

[ — = No samples collected]

Collection Point

Dieldrin Residues (PPM)

Sample

Water

Silt

No.

Post-

Pre-Treatment

Post-Treatment

Pre-Treatment

Treatment

Date

Amt

12-19-63 3-23-64

Date

Amt

12-19-63

1

Irving Park Pond,

Upper side

11-2

<.0O01

— <.0001

11-2

<.001

—

2

Sperry Creek, bank

of Horseshoe Bend

11-2

<.oooi

— <.O00I

11-2

.008

—

3

Kalamazoo River,

E. of bridge

10-31

.0002

<.0001 <.0001

10-31

><.001

<.001

4

Holmer Creek

at West River Rd

11-2

.0055

<.0001 <.O001

11-2

<.001

.002

5

Waubum Creek

—

—

<.0OOl <.0001

—

—

.003

6

Below Junction Kalamazoo

and B. C. Rivers

10-31

.0002

<.0001 <.0001

10-31

■<.001

.008

7

Kalamazoo River at

Country Club

<.0001

<.0001 <.0001

.009

<.0Ol

8

Harper Creek, before

Kalamazoo Rd.

10-31

<.0001

<.0O01 <.0001

10-31

.004

<.001

9

Goguac Lake, 116 Fern

10-31

.0002

— <.0001

10-31

.010

—

10

(a) Goguac Lake boathouse

(b) Vince Island by

10-31

.0006

— <.0001

Stone boathouse

10-31

.008

—

U

Kalamazoo River,

W. of bridge

10-31

.0006

<.0001 <.0001

10-31

1 <.001

<.001

12

Battle Creek, Elm St.

11-2

.0003

<.0001 <.000I

11-2

.006

<.001

13

(a) Battle Creek,

West Pony Ave.

11-2

.0003

<.001 <.0001

(b) Small Elm area at

bend in river

11-2

.11

.030

' Interferences made analysis impossible.

Vol. ],No. 4, March 1968

33

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