mM^ A STUDY OF THE BIOLUMINESCENCE OF A DEEP SCATTERING LAYER ORGANISM (EUPHAUSIA PACIFICA) IN MONTEREY BAY, CALIFORNIA Andrew Jerome Compton NAVAL POSIGiMOATE SCHOOL Monterey, California CBREPSKSD 1 /^^^^ A STUDY OF THE BIOLUMINESCENCE OF A DEEP SCATTERING LAYER ORGANISM (EUPHAUSIA PACIFICA) IN MONTEFlEY bay, CALIFORNIA by Andrev; Jerome Compton Thesis Advisor: Thesis Advisor: S.P. Tucker C.R. Dunlap, III March 197^ T159594 kppioxJtd {,01 puhtic fielejue.; diit/ubution untiriuXtd. A Study of the Biolumlnescence of a Deep Scattering Layer Organism (Euphausia pacifica) in Monterey Bay, California by Andrew Jerome ,j:^ompton Lieutenant Commander, United States Navy B.S., University of Miami, 1957 Submitted in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE IN OCEANOGRAPHY' from the NAVAL POSTGRADUATE SCHOOL March 197^ ABSTRACT A vertical migration tube (VMT) was designed and constructed as an Instrument to be used with a photomultl- pller light detector to make In situ mesopelaglc studies of deep scattering layer vertical migration organisms. Initial tests of the unit demonstrated the feasibility of its use in the marine environment. Three major problems of marine blolumlnescent studies using an underwater photomultiplier ' detector are resolved in part by the use of the VMT with this sensor. Mesopelaglc euphauslld crustaceans captured in the upper 100 meters of the water column at night decreased their blolumlnescent flash rates when lowered in the v/ater column and exposed primarily to pressure and temperature changes. There may be an Increase in euphauslld blolumlnes- cent flash rates when stimulated by other blolumlnescent organisms. Laboratory test equipment and laboratory methods were developed to permit quantitative measurements of euphauslld blolumlnescent output. Laboratory tests of Euphausia pacifica indicated a greater blolumlnescent response to a standard flash stimulus during midnight tests as opposed to noon tests. Laboratory tests of blolumlnescent activity during periods of moulting indicated greater than average response to a photoflash stimulus just prior to moulting and less than average response just after moulting. TABLE OF CONTENTS I. INTRODUCTION 9 II. NATURE OF THE PROBLEM AND OBJECTIVES 13 A. NATURE OF THE PROBLEM 13 1. Equipment 13 2. Methods 14 B. OBJECTIVES 15 III. INSTRUMENTATION 19 A. ELECTRONICS 19 B. VERTICAL MIGRATION TUBE 24 C. LABORATORY TEST EQUIPMENT 25 IV. PROCEDURES, TESTS AND RESULTS 31 A. AT SEA 31 1. General Procedures 31 2. At-Sea Tests and Results 32 a. Equipment Checkout 32 b. Ambient Light Measurements 33 0. VMT with Euphausilds (Excluding Ambient Light) 37 d. VMT with Euphausilds (Including Ambient Light) 41 B. LABORATORY 4l 1. General Procedures '^l 2. Laboratory Tests and Results ^5 a. Spontaneous Activity Test ^5 b. Stimulated Bioluminescent Activity Versus Ambient Light l^j c. stimulated Blolumlnescent Activity as a Function of Temperature J Moulting, and Time of Day 50 d. Sympathetic Blolumlnescent Response 90 V. DISCUSSION 92 A. AT-SEA RESULTS 92 B. LABORATORY RESULTS 97 C. GENERAL DISCUSSION AND FUTURE USE OF VMT 100 VI. CONCLUSION 103 APPENDIX A Chart of ACANIA Cruises 121 APPENDIX B Chart of Euphauslld Species Found at CalCOFI Station 3 122 APPENDIX C XBT at CalCOFI Station 3 123 APPENDIX D Spectral-sensitivity Characteristic of 1P21 (S-4 response) for a radiant flux from a tungsten source at 2870°K. 124 APPENDIX E Calibration Curves for 1P21 Tube 125 LIST OF REFERENCES 126 INITIAL DISTRIBUTION LIST 129 FORM DD 1473 131 LIST OF FIGURES 1. Map of Monterey Bay showing CalCOPI 1 Station, CalCOFI 3 Station, Hopkins Marine Station (HMS) and Naval Postgraduate School (NFS). (Depths In fathoms) l8 2. Spectral sensitivity characteristics and typical sensitivity and current amplification characteristics of 1P21 phototube. (From RCA Handbook of Photomultlpller Tubes) ■ 20 3. Photomultlpller tube In underwater housing 22 k. Circuit for photomultlpller light detector showing subsurface (A) and surface (B) units 23 5. Vertical Migration Tube with and without photomultlpller sensor attached 26 6. Euphauslid showing general terminology (After Mauchllne and Fisher, 1969) 27 7. Laboratory test apparatus showing sequence followed in testing euphausllds. A. Euphauslid in tube B. Euphauslid in test chamber C. Test chamber with euphauslid in reflector D. Apparatus in position E. Spontaneous test 29 8. Chart recording of dark current of photo- multlpller light detector at 100 meters depth 3^^ 9. Bioluminescence of environment at 25 meters (A), 50 meters (B), 75 meters (C), and 100 meters (D) 35 10. Bioluminescence of environment at 125 meters (A), 150 meters (B), 200 meters (C), and 250 meters (D) — 36 11. Bioluminescence of 10 euphausllds at 25 meters (A), 50 meters (B), 100 meters (C) and I50 meters (D) and 50 meters (E) and 25 meters (F) on return trip to surface 39 12. Continuous trace of bioluminescence of environment from 50 meters (top right) to 300 m.eters (lower right) and return to 50 meters (lower left). Read right to left. 8:00 PM '^O 13. Continuous trace of 10 euphausllds from 50 meters (right side) to 150 meters (left side). Not exposed to outside light. 9:00 PM ^2 14 . Continuous trace of 10 euphausllds from 50 meters to 125 meters (right to left). Exposed to outside light. 9:30 PM Hs 15. Spontaneous biolumlnescence test results ^6 16. Example of spontaneous activity of 15 euphausiids 48 17. Example of complete record (A) of stimulated euphausiid and example of almost (actually active for 2 more minutes) complete record (B) of stimulated euphausiid 52 18. First four tests of laboratory euphausiid #5. No. is day of month, n = noon test, m = midnight test 54 19. Second four tests of laboratory euphausiid #5. No. is day of month, n = noon test, m = midnight test 55 20-40. Daily activity charts for euphausiids. Day of month along upper abscissa. "n" implies noon test. "m" implies midnight test. "M" along lower abscissa implies moult formed. Flash rate in flashes per minute. Light output as area in cm2/2.5. Maximum amplitude in cm. Reaction time in minutes 57- 77 ^l-4§ . Individual euphausiid averages for parameters indicated (PR MLT implies "prior to moulting", AFT MLT implies "after moulting") 79- 86 49-50 . Group averages by temperature group for parameters indicated (PR MLT implies "prior to moulting", AFT MLT implies "after moulting") 87- 88 51. Chart of bioluminescent activity versus depth of environment and test groups of euphausiids for ACANIA cruises 74-14, 74-19 95 52. Bioluminescent records of environment and individual E_^ pacifica. a. Environment at 100 m depth; b. 10 euphausiids at 50 m in VMT; c,d. large euphausiid flashes at 100 m in VMT; e,f. laboratory recordings of light output of individual E. pacifica 96 LIST OF TABLES 1. Record of the measured parameters of the dally blolumlnescent activity of E. paclflca. 106 2. Averages of each measured parameter of the dally bloluminescence for each E_. paclflca. ll6 3. Averages of each measured parameter for each temperature group of E. paclflca. 120 ACKNOWLEDGEMENTS This study required work In several realms of science, many of which required specialized knowledge and facility which I did not possess. I wish to express special thanks to my two advisers; LCDR Calvin R. Dunlap for his help with the ecology of deep scattering layer organisms and many suggestions in laboratory and field work, to Professor Stevens P. Tucker for help with the electronics and suggestions in the field of light properties, and to both of them for numerous suggestions in the preparation of this thesis. Dr. George Evans offered suggestions and asked thoughtful questions concerning the endogenous rhythms of organisms . The captain, W. W. Reynolds, and crew of the ACANIA are among the finest group of men that I have had the privilege to work with. In eighteen cruises with them I received complete cooperation and help at all times. I also wish to thank Stanford University for the use of Hopkins Marine Station and Dave Brascher of that facility for help on numerous occasions. The NPGS Oceanography Department's Dana Mayberry and. the NPGS machine shop fabricated parts on many occasions. Finally, to my daughter, April, who spent many hours In a cold laboratory monitoring recordings of biolumlnescent activity I express my gratitude. I. INTRODUCTION Blolumlnescence Is the emission of light from living organisms as the result of internal oxidative changes. It is a characteristic of many pelagic marine organisms, yet it is one of their most poorly understood features (Tett and Kelly, 1973). Long recognized as a phenomenon in the surface waters of the ocean or in deep sea benthic fish (Beebe, 193^), the great extent of bioluminescent activity was not fully realized until the development of the photomultiplier light detector. This sensor, which can measure light intensities -7 2 to below 10 W/cm , has been used by several investigators (Clarke and V/ertheim, 1956; Clarke and Backus, 1956; Clarke and Hubbard, 1959; Kampa and Boden, 195^; Breslau and Edgerton, 1958; Clarke and Kelly, 1965; Neshyba, 1967; Rudykov, 1968) In ambient light and bioluminescent studies. These studies reveal the omnipresence of blolumlnescence in the oceans and suggest that it must have considerable ecological and behavioral significance. Light production is known to occur with a degree of certainty in ten phyla and about 35 orders of marine animals (Nicol, 1967). At least seven phyla contribute luminescence to the photic environment of the marine planktonic community (Boden and Kampa, 196^). The greatest development of luminescent organs in animals and the greatest proportion of luminescent forms are found in the mesopelagic regions of the oceans (Tett and Kelly, 1973). Many of the primary organisms thought to compose the acoustic "deep scattering layer" (DSL) are capable of biolumlnescence . The DSLs are a regular biological and acoustic feature of all the world's oceans. They ascend at sunset to near the sea surface and descend at sunrise to depths between 200 and 1000 meters. During this cycle they appear to follow selected isophots (Kampa and Boden, 195^). The organisms composing the DSL are among the most numerous on earth and comprise part of a great and complex pelagic ecology. They are basic to the oceanic food chain and are a primary source of volume reverberation of sound energy in the ocean. The acoustic volume scattering strength within the layers can be -70 to -80 db (at 2^^ kHz) but is variable within the layer. At lower frequencies the volume scattering strength is variable and unpredictable (Urick, 1967). These properties make the DSL an important consideration in fathometer trace interpretation and in military subsurface acoustic operations. The photomultiplier light detector was used by Clarke and co-v/orkers (Clarke and Wertheim, 1956; Clarke and Hubbard, 1959; Clarke and Breslau, i960) and by Kampa and Boden (195^) to measure the downwelling irradiance of sunlight at great depths. These investigators noticed the sensor also detected biolumlnescence and proceeded to investigate its distribution. Further v;ork by these and other investigators (Clarke and Backus, 1956; Boden and Kampa, 1957; Clarke and Kelly, 1965) has greatly increased our knov;ledge of the vertical distri- bution of luminescent pelagic animals. Interrelations 10 between vertical migration of the organisms, the rate and magnitude of luminescent flashing, and diurnal changes In the intensity with depth of the downwelllng sun- and sky- light have been documented, but the organisms responsible have not been Identified directly. A mldwater trawl used at sensor depths catches bioluml- nescence organisms, but the identity of a species that luminesces within range of the light sensor cannot be knov;n. Breslau and Edgerton (1958) and Bresiau, Clarke and Edgerton (1967) attempted to photograph the luminescent organisms. A photomultlpller detector was used which, upon being stimu- lated by the flash of a luminescent animal, triggered an electronic flash and camera combination. A few photographs were obtained of some coelenterates and crustaceans, but most of the photographs showed no recognizable organisms . The major objective of this study was to develop and use an in situ system for studying the bioluminescence of a given organism at depth. Work by Barham (1956) in Monterey Bay shov;ed that Euphausia paclfica, a luminescent euphausild, was a major organism of the bay's DSL. This crustacean is very probably the most common and widespread of the euphausild species in the North Pacific (Barham, 1956). and it plays a key role as a major link between the products of photo- synthesis and higher members of the food chain. E^ paclfica can be maintained in a laboratory for several weeks with minimum maintenance, and, therefore, prolonged laboratory tests are possible (Lasker and Theilacker, I965). 11 Investigators have generally neglected in situ testing of pelagic marine organisms. Hardy and Paton (19^7) conducted a series of In situ vertical migration tests on planktonlc animals using long glass cylinders at different levels in the sea. The cylinders had trap doors separating a number of Internal compartments. Messenger weights were used to control the doors and allow animals to move up or down within the cylinder in a given period of time under different conditions of ambient light and depth. The cylinder was sealed so that there was no exchange of water with the outside environment. Reviews of marine bioluminescence by Boden and Kampa (196^1) and Tett and Kelly (1973) do not mention in situ studies. 12 II. NATURE OF THE PROBLEM AND OBJECTIVES A. NATURE OF THE PROBLEM 1. Equipment Marine blolumlnescence falls In the visual light range, typically peaking between ^70nm and 500nm with total (all frequencies) power density at one meter distance ranging from lO"-^-"- to 2x10"''' yW/cm^ (Nlcol, 1962). At such low light levels the small Input light signal Is difficult to amplify with a good signal-to-nolse ratio except with a photomultiplier tube. A photomultiplier produces an electric current which is directly proportional to the amount of light falling upon the sensor and quasi-logarithmically proportional to the high voltage across the tube. The spectral sensitivity depends mainly upon the composition of the photoreceptlve surface, and the amplification depends upon the supply voltage and the geometry of the internal diodes . "Voltage control circuitry (to lower the voltage across the electrodes as the current increases) or a system of stops and filters (to control the light entering) must be used to achieve a large dynamic light range. The use of a sensitive, high voltage instrument in the marine environment poses some design problems. Sensors designed for in situ use are a compromise between sensitivity, available components, and the need for rugged construction (Tett and Kelly, 1973). 13 The photomultlpller light detector and associated components must be protected by a pressure-resistant housing and normally connect with the ship via a long electric cable. A photomultiplier tube operates from a high voltage DC power supply, typically 1000 VDC or higher. This must be sent down the cable to the sensor from a surface supply or carried in situ in the form of a battery pack or local high voltage supply. The instruments present difficulties in maintenance because of high input voltages, low current sensitivity, and high photomultiplier output impedance, but these can be overcome by proper design. 2. Methods Once a light detector is in the ocean the experimental techniques employed become of paramount importance. The sensor is suspended on a cable and, except in dead calm conditions, experiences a vertical oscillation which provides an artificial stimulus to the organisms and causes a biolumi- nescent response (Mauchline and Fisher, 1969). The distance and the angle from the organism to the sensor are unknown, and, therefore, the absolute light output of the organisms cannot be determined. The greatest failing has been the Inability of the investigator to identify the organism producing the light. Attempts to solve these problems have not met with success, and they remain largely unresolved (Mauchline and Fisher, 1969; Tett and Kelly, 1973). Ih B. OBJECTIVES The primary goal of this Investigation was to attempt to circumvent these problems with new methods and equipment and, possibly, to extend the present limited knowledge of marine blolumlnescence . A second objective was to attempt to relate the results (or possible future results) to military operations . The great prevalence of bloluminescent organisms in pelagic communities (Nicol, 196?) and the increase of bloluminescent activity recorded within the DSL (Boden and Kampa, 1957) indicate that blolumlnescence may be an important parameter that could be used to locate the DSL. The use of a passive light detector versus an active sonar to locate an acoustic scattering layer would give a submarine a great advantage in trying to remain undetected. Brown (1970) examined the possibility of detecting submerged submarines from the blolumlnescence Induced by their passage through biolumlnescent-active waters. The depth at which a submarine may be detected in this manner from air depends upon three environmental factors: the luminance of the v/ater background, the luminance of the blolumlnescence excited by the submarine surface, and the attenuation of the bloluminescent light by the water in the path between the submarine and the observer (Brown, 1970). The most uncertain parameter is the brightness of the induced blolumlnescence. Brown (1970) predicted detection to 120 meters depth with the aid of a light amplification device 15 in clear ocean water on a dark night. This study indicates that more in situ research is needed. The present study was conducted because it was felt that extrapolation of the results of laboratory testing of animals of the deep ocean is of limited validity, i.e. many of the results obtained in the laboratory are suspect due to the artificial environment. Once an animal is removed from its natural environment and placed in an artificial one there is a 'zoo-effect'. This is the change in the animal's responses and behavior effected by its laboratory environment. The environmental changes may be obvious changes, such as the physical changes in the ocean which DSL organisms are subjected to in diurnal vertical migration and which are difficult to reproduce in a laboratory. They may also be subtle and involve an environmental mechanism which triggers a circadian rhythm. Since in most cases the natural responses of the animal are not known (usually that is what we are trying to determine) the extent of the 'zoo-effect' is unknown. An in situ approach is therefore necessary for many experimental studies, and concurrent in situ and laboratory studies may be even more valuable. The following specific objectives vjere set: 1. Capture, identify and maintain Euphausla pacifica in a laboratory for experimental testing; 2. Design and build: a. An apparatus having a photomultipller detector to measure and record bioluminescence ; 16 b. Laboratory and In situ test equipment to obtain quantitative measures of light output by individual E. pacifica; c. Make at-sea in situ tests of DSL organisms (specifically E_^ pacifica) from R/V ACANIA (the Naval Postgraduate School's research vessel) in Monterey Bay (Figure 1, Station 3); and d. Supplement the at-sea tests with laboratory tests of blolumlnescence from E_^ pacifica against such parameters as temperature, time, moulting frequency, oxygen, pH, and light . 17 36" 55'N 45" — 3 6°3 5'N 122''00'W 50' Figure 1. Man of Monterey Bay showing CalCOFI Station 1, CaicOFI Station 3, Hopkins Marine Station (HMS) and Naval Postgraduate School (MPS). (Depths in fathoms.) 18 III. INSTRUMENTATION A. ELECTRONICS Photomultlpller light detectors for at-sea bloluminescence studies were initially designed and developed independently by Clarke and his co-workers and by Boden and Kampa in the mid 1950's (Tett and Kelly, 1973). The basic circuit components are a photomultiplier tube, a regulated high voltage power supply and a recording or monitoring device. In this study a RCA 93IA photomultiplier tube was used initially and later replaced by a 1P21 tube. The tubes are identical, except the 1P21 has a luminous sensitivity about four times greater. The 93IA and 1P21 have a maximum spectral response at about o 4000 A (Figure 2). Thus bloluminescence (470nm to 500nm) occurs at the 705^ to 83^ relative sensitivity portion on the response curves of these detectors. The high voltage power supply may be located on deck or in situ. A shipboard regulated pov;er supply which could be adjusted manually in one-, ten-, or one hundred-volt steps up to 1200VDC was tried initially. This allowed the use of a much simpler in situ unit, as only the photomultiplier tube had to be encased in an underwater housing. However, this required a cable capable of carrying high voltages (1000- 1200 VDC) to depths of several hundred meters. Later the underwater housing was enlarged, and a high voltage battery pack was made from four 300 VDC batteries. A small operational am.plifier was placed in the underwater 19 TYPICAL SENSITIVITY AND CURRENT AMPLIFICATION CHARACTERISTICS SUPPLl en CEE AND e 4 ? WOO- X a ■g ' N 4 hJ K D 0- £ lOO r VOI TftiT in ACRO'S VOtlAGf Div ^EFN CAIHOPF A,\r DTNO: 1. rjj.l; 1 Dim:. DYNoor sia&f , 4no i/iO Of t ANODE. IflfB /I' 0 BF H fRO f E .Etr ViD'NG l/.O „,_ _- _- — ;^ ~ 7~ 1 : ' / 1 / / i r ' / ^ - "t" f 6 -cV jf j¥ ,.v / / €-. / V / ^_ g 6 o O 4 Z UJ £ 5 ' Si 2 Z kj en 1- • y— v -^ ^^ 1 h — _-_! 1 L- e / y — - --- ~ — e / / l^ — " / J' / , '' / — - - w / ^^-^ " ."_ ""."I — -'_ " ":. __L "~ R 4 J 0.1 f / — - --■ 6 [y- — -- ^ SPECTRAL-SENSITIVITY CHARACTERISTIC OF PHOTOTUBE HAVING S-4 RESPONSE FOR EQUAL VALUES OF RADIANT FLUX AT ALL WAVELENGTHS ' ■' « ' 1000 ^ ' -ISO. ANOOE - lO-CATHODF JUI'P^Y VOllS It I lliLtiiiLiS-i: 5000 7000 9000 WAVELFNG1 H— ANGSTROMS 1 1000 Figure 2. Spectral sensitivity characteristics and typical sensitivity and current amplification characteris- tics of 1P21 phototube. (From RCA 'data sheets 92CM-'61^2R9 and 92CK-61i5UR$, respectively. 20 housing along with the battery and a deck controlled on-off relay to apply the high voltage across the photomultlpller tube (Figure 3). The deck units consisted of an attenuator and zero offset unit, a Hewlett Packard model 680 five-inch strip chart recorder having a frequency response of one-half second full scale, and a small DC power supply to operate the on-off relay (Figure ^). The in situ detector unit. was suspended on R/V ACANIA's four-conductor armored hydrographic cable. Slip rings allowed measurements to be performed while raising or lowering the sensor. Calibration of the photomultlpller tube was accomplished with a Gamma Model 220-lA standard light source of 100 in a photographic dark room . The sensor was placed at a distance of 57 inches from a standard source rated at a color temperature of 285^ i 50°K and at 100 footcandles at a distance of one foot. The spectral-sensitivity characteristics of the 1P21 photomultlpller tube (S-4 response) for a radiant flux from a tungsten source at 2870°K are given in Appendix D. o o This peaks at about 5100 A, fairly near the il700 to 5000 A for bioluminescence in the ocean. At a distance of 57 inches the amount of light from the source is reduced to ^.H3 footcandles . Neutral density filters were used in steps of 1.0 to reduce the light to the sensor. The high voltage to the 1P21 tube was Increased in 100 VDC steps from 800 VDC to 1200 VDC . The calibration curves are depicted in Appendix E. The signal in millivolts to the recorder (Hewlett Packard Model 680) 21 J 1^ s " 1" J a 01 :3 o Xi u ■P cd & U (U •o c •H (U :i ?H 0) •H rH P. •H -P iH O 4^ O Xi CO u bO •H 22 W w <\J ir» rH CM rH II ir tH fM VC. K iH CM P-i fH 01 C>,|l" /f^ ^^ pT K K ■W't: (li ■p h: ^ v^^ cr; f— I — rH Hi. «J C^)l <- r\ \ >r V Ml" -p ^ 3 rH rH D, fS B^ CO o a :3 oa CO bO C ■H O w o ■p o (1) p -a •p bO •H iH 0) P. •H • -P W rH P :3 -H e c o ::! p o '-- ^ CQ Jh (D O O Cm ro Cm -P Jh •H ;3 us w o •H C o ro (1) bO •H 23 Is plotted versus the illumination at the window of the detector in footcandles. No corrections were made to take into account the fact that the Fresnel reflection coefficient for normal incidence for the air-plexiglass window surface of the detector as used during calibration and laboratory work differs from that for water-plexiglass surface during the in situ measurement. /np - n \2 For the air-plexiglass surface, r.p = 1 :j: I , and for the water-plexiglass surface, r,,c = 1 T , where n-r, = 1.49 is the refractive index WP Vnp + n J ' P for plexiglass, Hp = 1.3^ is that for seawater, and % = 1 is that for air, all for the sodium D lines. Thus, since the calibration v;as performed in air, the in situ recorder readings should be multiplied by a factor ^^ ? r 1 - (.r_^p - r^p; = 1.038 to obtain the true in situ light values. In the present study no attempt has been made to make this correction as it is small compared with the other experimental uncertainties. B. VERTICAL MIGRATION TUBE (VMT) The major problems of artificial stimulation of the organisms in the ocean by the equipment, the unknown distance from sensor to organism, and the inability to Identify the exact organism or oganlsms emitting light was approached by designing and constructing an underwater retaining tube. This tube holds the organism (or organisms) being tested at a known distance with minimal mechanical stimulation. 24 Designated as a vertical migration tube (VMT) It was designed to contain small organisms of euphauslld size (12-25- mm length) within the field of view of the sensor and yet allow the organisms to swim freely within a restricted area (Figure 5). Access holes at either end of the tube allow pressure equalization, while temperature equalization occurs through the plexiglass. The tube Is suspended from the hydrographlc cable with the photomultlpller light detector attached (Figure 5) and then lowered Into the ocean. Light can be shut out from the V>1T and the detector by placing a black plastic curtain around them. Retaining screens, which hold the organisms at any desired position along the VMT, allow free flow of water. Access to the VT-IT Is through removable base plates. Two supporting rods of 3/8" aluminum allow a heavy stabilizing weight to be attached. C. LABORATORY TEST EQUIPMENT Initial attempts at laboratory testing of the blolumi- nescent light output of an individual euphauslld v;ere indeterminate when methods previously developed (Tett, 1972) were used. This was due to the continually changing aspect and range presented to the sensor by the freely swimming euphauslld, the photophores (luminescent organs) of which are located laterally along its abdominal segments (Figure 6). V/hen placed in a small beaker, stimulated by electronic flash (Hardy and Kay, 196^]), and placed over the photomultlpller 25 Cable ^^ >9 T X X }*-z- BASE PLATE ACCESS HOLE ' l/i|" DIAMETER SENSOR IN HOUSING ' *^--. SUPPORTING ROD i^"/OD PLEXIGLASS TUBE h ACCESS HOIE l/i|° DIAICTELR ht'H RETAINING SCREENS ST.ABILJZING VEIGHT Figure 5. Vertic. 1 Migration Tube with and wi^jhout photOi.-ultiplier sensor attached. 26 U (1) X! m •H ■O C cd o C •H iH X! o :3 cd +i bO O H O •H e (U A^- 1 ! : 1 ■ ! ! 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O 4J O a o 0) o (H Cm O Td c 0) cti o c ^^ (D o O •^-^ W Q) ra c ^ •H 0) e p> :^ flj iH S O •H LO CQ t^- (D •H 35 t^t 1 ' ' ■ ^-H -i-i-: 1 .; ; pi-iJ >_ ^.^ ' . , L ' ' ] 1 -;-H-^ .111 r :_ -, -i-l-L W: -*-T •^ -^- -" * " — ' — £Z-J — ■ — - J J '/^ -^-f-r+" T- __—_*--,_ .^- *-_._,- ^ -- T-^ ■ 1 ->ll.-. ■——r- -; — - _ — — -r^ 5--^^" — - ._ — — ..,-J — ^ ^— ^ - : ^ lL ZT'.'T -.1 — . . ..... .i_j — — .. -•- -_ ~^-- - — — f- "— - ----- :.".". i:L -•-n-t- .-.; — - - - - — r- J. — .-. rr" — 1 — ~ 7:--; — - — - _^ — -^ — M : — U=q — " 1 ■ -- : -- — , 1 TTT ' T- — -t — ,. -— -- - — ' _ -- — — — —[ — - . — '■ — J TV _: — - — . ^- r - - 1 -— '- — — V-- < /^ I- J -:c ._... .-"....1. — - — - 1 -- - i 5 ^ 1 — - 1 — 11""ZT1 . .__._ "1 -::.r|Ml — --— ..::: ^ — , : 1 i'VJ 1 ::: :-Tf.~ ■ — — r-r- - — . _- . zr." t:^ 1 ■ ^-t-i— _ -^ — --' ; "^:z . j .... ■ ~7 " ""' ■*'. -— " '-'-'-'■ "-- — — - ..; ,• — - — : -r- -i-L- ~m — ^-f xq Tii ■T- T-* —- rr ■t-f-^ _ 1 ;_ —i- -+-h4 ^■, ." TTTT ■ f ■ ■ -■U4 -• - t » iilf -,-i-f-i. -^ -*-*-;-- : II — - [rr ni — ._ — .-1 ■-1 ■r- 1 ; ■ /5 -^ ■•-ALU • i •■ -■ ' — r -• ' f « ■ ■ ■ T ^414- -■ r-TT 01-^ ' ■ ■ ' . , 1 < -i-f-t-f- ' ' ' 1 TITt -V-^ .! i 1 . -T-r--- ^-: h : : . T 1 ; .4— -1 t ■ -T-T Ti , :_ +Frr i ■ i , ? — ^ — ^ - »— 1— ~r-\ ±i:rr _,-J L_ .* . . 1 • i ■ ■ • -r-t- - . i-i_ 1 ' , '- 1 1 ' _i ■ ■- ■ 1 ! ■ ; 1 ' ■ 4x4' -T-T 1 1 4J-;: ' '"i ^^ :^ ! • I r Ml! 1 t 1 ■ 1 1 1 ' ill! -5- n- 4- ~r T -♦ ■ -t- -t T -rrrr ! 1 1 ■ ■ :riu: .[ITT -t-T r-r 1 ' ' ■' — t m - -4- -4- CQ •p .e o LTV W P-i ^ 0 0 -p 0 G) • • B 0 >-| LP* C\J r-l ■P ^-^ Cti Q P C W 0) i^ B 0 C LTV m (M Cm ■d 0 C cti 0) 0 ^-v c 0 OJ ^^ 0 w to (D f-, c 0) •H +i e 0) :3 e iH 0 0 •H 0 CQ CM o bO •H 36 c. WIT with Euphausllds (Excluding Ambient Light) Once the vertical profile of blolumlnescence had been recorded to a predetermined depth, a net haul was made to collect organisms to test In the VMT. The VMT was attached to the electric hydrographlc wire and the light detector to the VMT (Figure 5). The VMT was then carefully filled with surface water to prevent the formation of small bubbles, which can attach to the euphauslld appendages and hinder movement. The first retaining screen was positioned, ten euphausllds were introduced, and then the second screen placed into position. The end plate was then fastened into position, and the sensor and VMT partially wrapped and taped with black plastic to screen out ambient light. The unit 'was then lowered to 50 meters, where it was kept for 20-25 minutes to allow battery temperature compensation and to permit the euphausllds to become acclimatized. Stimulation due to handling and exposure to light will cause initial blolumlnescence in the organisms. The role of blolumlnescence in the marine environment and how physical and environmental factors affect light output are largely unknown. Hardy and Kay (1964) and Tett (1972) have shown that mechanical agitation and bright light can stimulate blclumlnescent activity in labora- tory tests of euphausllds. After acclimatization the Initial tests were repeated v;lth the sensor detecting only the blolumlnescence of the ten euphausllds. The euphausllds were subjected 37 primarily to pressure and temperature changes In the water column. Other parameters could affect this activity however. There are comparatively large changes in oxygen, pH, carbon dioxide, salinity, and both phosphorus and nitrogen (in the form of nutrients) in the upper oceanic waters (Home, 1969). It is felt that these changes were probably attenuated due to the small size of the 0.25-in diameter access holes, the relatively large internal volume of water within the VMT, and the distance from the access holes to the test area within the tube (Figure 5). The bioluminescent record of the euphausiids was similar in profile to the record of environmental biolumi- nescence made an hour earlier on the same cruise. The flash frequency of bioluminescence again decreases with increased depth in the water column. Here, however, the euphausiids are physically restrained to the same area of the VMT and the decrease in bioluminescence must be due to less activity by the same number of organisms. The bioluminescent activity increased as the sensor and VMT were raised (Figure 11). A second series of tests starting three hours earlier in the evening was conducted on ACANIA cruise 7^-19. The bioluminescent activity of the environment did not show the marked decrease with depth evident on the first test. A • continuous trace is used to depict this in Figure 12. It is felt that the increased bioluminescence at depth compared to that for the previous test (ACANIA cruise Y^-l^J) was due to Incomplete vertical migration of the DSLs due to the earlier 38 -':■..."■'-[ " :: : .... ■■■■]■■ ■ 1 Vl I * : : ... 1 ... - - ! - . - . -_ . ^ :-l: :::->•; ^• i£r i:"i; i ■1 ■ I. • ; ■ i: ". . ' ' ' . - 1 ; • 1 1 ". '1 ' ■ 4 "- \ " ■ i " H 1. /^ - ■-: - '{- ^ ': ■■! . - 1 i - ---i- -- zzi':i- ::-.:] .'--"-,. ■ ■ ■ -■-': 7"' .rm ^ ^ u, 1 r .r;:!-^ -I -:: ;;;; 1 " r . I I ; r^- ■ ■■':. ^ b:;l T ■ -^:j 1 :; - r , r.:: 1 ; :1^ ■ ' !- 1 ■ \j , ■ \ * Ph to U Qi m -P (U m E in in OJ U 0) -O -p C 0) cri B o W in 0\ w -"-^ ^H < O ^^ ■P a) w e u d) o -p in 0 fc: -p LPv C\J -a -P Kl a • *\ w ,-^ -0 Q •H v_^ •H W ra 13 fn cti 0) x: +J • Q< (U a) ::! td o •H u ■p x: • bO+J •H ^ ^ bO ^-' •H iH W ^ (1) 0 •a -P •H 0) Kl a P ;:! o o U~\ o B p o U -d ' 1 ■ i ' M y 1 1 1 n , ' i . ' * , • 1 ■ 1 i ' ; ^11 :, . ._;. .i .. ._!- - , Am-'- kl^-^-^- 1 , ; ' J — . -^. '=) T| 1 _i„ *" ■ ■ -U-*.^ 1^ 1 . i --,---— .. ! 1 1., — .— 1-4— i— „l^_i.j_ — A— j 1 ' ' ! -1- U u — --r--— - --- — ^- -^ — ;■-?- ..j-._ --/ I'M , , ' ' 1 J — --- - 1 • ,t j ^^ "-- 1 , ■ •^ 1 ' ■ , 1 "J . i i i 1 , 1 ; : 1 . 1 ■ ^^ Pi ^ ■ ' ■ 1 , ! ; 1 ; 1 1 1 III ^ ■ 1 i 1 ; , J ' \ 1 ' ' - ■ - '— — J ... .... -T^ ::•:.: - i ' ; ..-._. R _. ^ - ^ . ..... _. \ i • J i ____ •- — - - ;-f--— ^ J V .: 1 ■ - ■ \ \ 1 -^- [iV^-1 ■ 1 1 ' 1 1 ; . , 1 : ■ ' 1 i 1 . Mil ' • \ • 1 , ■ i M 1 i i 1 1 • ' ' i ' J ; ' M i ■ 1 : ! ■ , 1 1 i ~1 - - -1- - - 1 ' ■ ' ' 7 ' 1 1 -" , ! ' ! 1 1 ■ , , ' 1 , . W I ' i ^ 1 1 . i : M 1 ! ' ' , ! 1 j 1 ' . : 1 ■ i i "^ ■ 1 : ! 1 ' : ' ' 1 i i 1 1 ' 1 ' , 1 1 ; 1 1 : j ; ! 1 J , , 1 1 ■ i 1 1 ; ill ! ; 1 1 1 1 1 1 ' ! ; i ' 1 1 ' . 1 • ■ 1 ' 1 1 ill!, • i : ' ' li ! 1 ' 1 J ' : ! i . . : 1 ! : i ! i ' 1 '; ■ ■ ■ ' 1 ; ; 1 i i i i ' 1 j I'M ^^J < "''"rr : -_-^.. — 1 1 1 ill! f-^r 1 1 1 1 1 ■ ] I ' Ll'-i-T "iTLXX' 1 i 1 1 i ' ■ j M ' -i- ' 1 : ' I i < ■1 1 i 1 1 1 5^ "T' '"^ 1 1 ■ ' \ ' - M • ■ ■ ' i ' , 1 1 ; ' ■ ; ; 1 • 1 ! ! ' ! ' 1 ' , u^^ U-^ I. !_.!._ 1 , 1 ! i i : 1 , ' ; i !i II' ' ! < If ' 1 i ! i , Ml: ' ' ! ' ■I ._.. 1 : , , 1 1 1 i ! ! . ! ; 1 , . I I ill' ! • ' ■ i i ' 1 > ' 1 ' i ; 1 ' '1,1 i i 1 i i 1 : I 1 , 1 : ■ 1 : ■ ; ' ' 1 1 ' ■ : ! ■ J_' ' CI \ ■ . '■ '• ; •Hi 1 * ' ' ^ ^• . ,_ .. — . — OJ- ■ - 1 ! < 1 1 ! 1 I : 1 ; 1 . ■ 1 ; 1'— - ' ■ ,-' - r- , ' i ; . ' 1 ' ' , ' 1 ! -J ' ' ' ..-:- ,, — - — T— ■— t J 1 ' ■ -^- ALU 01 — >- -r: ' : 1 ' ■ 1 ' ; ~\\ ! . _-i— ; . ^y -i-i ,-L- :■ .xJ,T : ■ 1 I 1 . ' t [ , ■ n ' 1 i ' ■ill" i~^i '• i i 1 i i ' 1 111! i 1 ! 1 111 'II' 1 ' T , ' M ' 1 Si bO •H U (0 u , -p >. •H ■JJ OOfrOr^ > •H •^ -1^ ■P •,H •H c; -P > ■ tn O •H a ■JJ ■i-^ ■fh P •H trl o r-l <1) c to x: f^ id iJ6 four and five (12°C) were tested for periods of 25 and 34 hours respectively. They showed more variation (less similarity) and more inactive time than the other groups (Figure 15). They matched activity only between 1530 and 1700 hours. Group four showed internal matching from 150O to 1700 hours. In evaluating the records no attempt was made to count the individual flashes, as low amplitude and slov; chart speed (four inches per hour) compressed the individual flashes together on the record. The results were evaluated subjec- tively as "heavy activity" if the flashes were compressed into a solid record (Figure I6), "light activity" if the base line was evident between flashes, and "no activity" if only the base line was evident. The results were inconclusive when evaluated for a 12 or 24 hour cyclic rhythm. b. Stimulated Bioluminescent Activity Versus Ambient Light Stimulation of euphausiids in the laboratory to excite a bioluminescent response has been accomplished most successfully with a photoflash unit (Boden and Kampa, 1964) . This method was adopted as a standard stimulus for the present study. The euphausiids were tested individually by removing each in turn from its one-quart laboratory container, placing it in a 60-ml glass beaker to which had been added 40 ml of water from the laboratory container, and flashing the beaker with the photoflash unit at a 10-cm distance when the euphausiid approached the side nearest the flash unit. 47 Figure 16. Example of spontaneous activity of 15 euphausllds ^8 The beaker was then immediately placed over the photo- multiplier sensor, which was shielded from outside light by placing a black cup over the beaker and sensor; the room light was turned off; and high voltage was applied to the sensor. An initial qualitative test of bioluminescent activity of groups of euphauslids kept in different light environments was made to determine if am.bient light levels would affect their light output. This also served as a test for the laboratory methods and resulted in the development of nev7 procedures and equipment. It was felt after this test that techniques used up to this point were inadequate (see Chapter III.C) to evaluate quantitatively this parameter (Hardy and Kay, 1964; Tett, 1972). A two-week test was run with twenty-five euphauslids which had responded positively to photoflash stimulus. They were divided into four groups as f ollov/s : ten euphauslids were placed in a completely dark environment, five were placed in a "bright" light environment (a 60 W, 1000-hour bulb at two feet), five were placed in a "dim" environment (estimated to be about 10 -^ yW/cm — measured with a 931A phototube which was not calibrated absolutely, but was compared for relative sensitivity to the 1P21), and five were placed In a 12 hour alternating "dark" (night) and "dim" (daytime) environment. The ten euphauslids in the "dark" environment were tested once each day at l800 and 49 also checked for the presence of moults. The other three groups were tested once every third day at 1700. Subjective analysis of the records indicated that the bright light may have affected bioluminescent activity of that group as its bloluminescence fell off more quickly with time than that of the other three groups . There v/ere no observable differences among the other three. The bright light group also had a higher mortality rate, i.e. all were dead at the end of two weeks versus a total of six dead out of twenty for the other three groups. A low light level environment was adopted for maintaining euphausiids for future tests. This was convenient for laboratory work and did not require abrupt light changes for feeding and change of water. No obvious relationship betv/een moulting and bioluminescent activity was noted. c. Stimulated Bioluminescent Activity as a Function of Temperature, Moulting and Time of Day Tests based upon the experience gained from the previous ones were set up to check bioluminescent activity against two temperature controls, time of day, and mioulting. Three groups of five euphausiids each were established after an initial test for positive bioluminescent response to photoflash stimulation. Euphausiids not bioluminescing within a four-minute time lapse after stimulation were not used. Group one (euphausiids #l-//5) were kept in a 12°C constant temperature bath, group two (euphausiids 50 #6-#10) were kept In baths which were alternated from 7°C to 12°C and from 12°C to 7°C at Intervals of about 12 hours; and group three (euphauslids #11-#15) were kept in a 7°C bath. Group two was changed from 7°C to 12°C v;ater just prior to testing at midnight and from 12°C to 7°C v;ater just prior to testing at noon. Tests were made daily from 1000-1400 and 2200-0200 for periods of up to 23 days. Moults found, time of test, water temperature, and an indication of bioluminescent activity were noted for each euphausiid tested. A six-minute record of the bioluminescent activity was made to determine light amplitude, flash frequency, total light output and reaction time from stimulus to start of bioluminescent activity (Table I). All groups were kept in a low light level environment with no disturbance except that resulting from testing. Earlier tests made as part of the present study indicated that a six-minute record of the activity of the stimulated euphausiids was long enough to yield a good indication of the bioluminescent activity of the organism and not so long as to make the testing of fifteen euphausiids unmanageable. Some euphausiids displayed activity for as long as seventeen minutes, some for as few as two minutes. Eight minutes was the average period of activity. Figure 17 is an example of complete records of stimulated euphausiids which also Illustrate these differences in activity. During each test-period euphausiids were taken, one at a time and in their individual containers, to a 51 6 5 4 Time in minutes Figure 17. Example of complete record (A) of stimulated euphausiid, and example of almost (actually active for 2 more minutes) complete record (B) of stimulated euphausiid. 52 separate dark room. A glass tube was used to capture and transfer each euphausild to the test chamber following the procedures described In Chapter III. The Bauer photoflash unit was used to deliver a standard stimulus by flashing the dorsal aspect of the euphausild at a distance of approximately 10 cm. The test chamber was then Inserted into the reflector which was positioned over the sensor aperture. A rubber seal between the reflector and the sensor housing and between the test chamber and reflector guarded against stray light. A black plastic hood was also used over the test apparatus, and the room lights were turned off. Because the average time from application of stimulus to the start of recording was 15-20 seconds, a 0.3-niinute correction was added to each recorded test start time in Table I. A 50-mv full scale recorder setting vms used. This was a compromise between a more sensitive 5-niv setting, which would have detected lower level light activity, and a 100-mv setting which would have kept all readings on scale. A complete series of recordings for one specim.en is displayed in Figures l8 and 19. The records show the change in the activity of the euphausild from test to test and illustrate the type of laboratory record made for each euphausild. The record for each euphausild was evaluated to determine total light output, flash frequency, maximum amplitude and reaction time. The total light output (area under each curve) was evaluated with a compensating polar 53 3 2 Time in riinutes Figure 18. First four tests of laboratory euphauslid #5 No. is day of month, n = noon test, m = midnight test. 5^ Time in ndnutes Figure 19. Second four tests of laboratory euphauslid #5. No. is day of month, n = noon test, m = midnlKht test. 55 planlmeter (Dletzgen Model D-I806). This area in square centimeters was then divided by 2.5 to normalize area to the other scaled parameters. Flash rate was found and the average rate per minute calculated. Only deflections greater than 2 mm in height were counted as flashes. In cases where the flash rate was Irregular the total number of flashes was counted and divided by the total elapsed time to obtain the number of flashes per minute. If the flash rate appeared to be regular, two one-minute portions of the record were counted and averaged. The maximum amplitude or deviation above the background level was measured and recorded in centimeters deflection. Occasionally the light output would be high enough to drive the recorder pen off scale. This was not compensated for in area or amplitude measurements, as only a relative indication of euphausiid activity was sought. The activity of each euphausiid was then charted for each parameter at each test period from the results of the above evaluations. The date and time of test are along the upper abscissa (Figures 20-40) . A noon test is indicated by "n" and a midnight test by an "m" . Moulting is Indicated along the lower abscissa by an "M". The measured param.eters are listed along the right ordinate. A break in the graph indicates that more than 24 hours have passed since the previous test. 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CU U CO E 0 cd CD ' -H Cm (D p o\ ^ 13 ^ = P cd C rS rH -H cd :: :3 CO S 0 0 cd •H B c: Cm • P -rH •H p CO Zi 0 w OJ Q (D Oj (D •H P fi a p rH d -H a 0 P • c E w 0 •H P C 0 ^ 0 fn C Cd hO-H 0 to ^ p Cm w to kJ] CJ (D •H Cd P -H 0 Q) fn rH to • K Oj Pj X3 0) ^ E cd p 0 tH :3 • U C B >;> = (D -H 0 P C S e •H = 0 C > rH fn -H •H 2-H cd ^ -H rH 0 CO :-\ •H W = cd a. cd £! s rH e Q cd :: Cm cd en , w • 0 cO 0) "O D -H cfl C e rH -H cti r :3 to s 0 0 cd •H B a Ch 4J -H •H -P to d 0 to oj a, Qj Cti (U -H P> S a. -P rH ::s -H a 0 -p • c B w 0 -H -p d 0 4:^ 0 fn C Cti bO-H 0 CQ -H -P Cm CO CO J 0 (D -H Cd -P -H 0 0 fn iH CO • K Cti CUX3 (D XJ S Cti -P 0 ^H US • U C B >>= OJ -H 0 -P C S S •H r 0 C > rH J-l -H •H (D 4J • hO P< to 0 cti C 'O oj CO 0 CO ::S (0 rH 0) -P • >3 -H Ct3 £ -H 1-1 0 CO rH •H CO = cd Q. cd X3 S H E Q cd = Cd cdS £; -p ^ -p x: c wx: ' O -H CO LPv E C Cd • xi iH C\J (U. ^ fe \ o S CM E >5 01 • o «J 0 'Ci Q -H Q) C iH E -H • a ?H to • e O cd 5 = OJ -H O -p t: ^ E •H = o d > rH Jh -H •H (U -P • M a 0) o Cd C 'O cd CO o to ::s w rH (U -P >>-H cd ^ -H H O to cH •rH W = cd a Cd JD s rH e Q Cd = Cm cd m ro (U fn :3 w fe 70 3WI1* o in t n M «- dwv<;j? 5 *- o m in in vanvao o in o o o o D3y doo o in o o n o CM o u > w • o Cti 0 tj Q -H 0 C rH E -H • Q fn W • s O Oj 0 rH ^ Cm 0 -P iH fn :3 =fc r ■P oj a s r-i -H kJ r 3 to s o o d •H E C «M • P> -H •H ■P [0 ::i O W 0 a 0 a 0 •H -P E u -P rH :3 ^ a o -p • c E w o •H -P G o x: o ?H a nj hO-rH o CO -rH -P ^ w to hj o (D •H cd -P -H O 0 ^ rH CO • K erf axi 0 x; E nj -P O -H ^ • . fn C E >:,= d) -H O -P G 5 E •H r: o t: > rH fn -H •H 0 •P • bO D. 0 o cd C 'd cti w o to ::S w rH 0 +3 >>-H CC ^ -H ■H O to rH •H W = cd a Cti £) S rH E Q cti = Cm Oj • oa 0 u :i hO 71 vaavno D3iJ Jo o in in n in « in in 72 V3a yo in o in o m fO M in in in o o o o O O . ^ ::! S M • C ;3 C -P •H B o w •H iH 0 05S x: -p ^ -p ^ c box: ♦ O -H to LO e c c;j • ■a t-\ CM Cm -H fe\ o B CM B >> to • o — 03 (U 'O 1) c rH B -H • a fn to • e O 03 q; OJ tH •H •H -P to d O w (D a (U OJ 0) •H -P S P< -p rH :i •H cu o -P • C S W O •H -P c O x: O !^ C cd hO •H O to -H -P Cm W to ^q O 0) •H 03 P -H CJ (D !^ rH to • (t; Cti a£l CD ^ e o3 -P O -H S • , Sh C S >3 = 0) -H o ■P C ^ B •H = o C > rH ^ •rl •H (D -P • hO Q< 0 O «j C t:! oj w o to d w rH 0) p >i-H 03 ^ •H rH O CO H •H W :; 03 a cd ^ S rH B O 03 C Cm o3 M3 on (U U :i bO •H Ph 73 3Wll*o in t m M »- dwv<;2 in o in in in vanvno o ■n o o m O Dan joo o O in o ■<*■ o o O ?^ (U a. a 3 B M • c :3 C -P -H S O M -H >H 0 i w • 0 Cti (0 Td Q -H (U G rH S -H • D. f^ Kl • a 0 nj (1> 00 ^ >= rH f-l -H •H (D -P ' W iX (U 0 nj C TS nj w 0 w d M rH (D -P >> -H cti x; -H rH 0 W H •H W = 03 a Oj X3 S rH S Q oi r (h oj • oo 0) fn d bO •H fo n awii*'0 in fO c< dwv<;;[! in o in in in vaavno o o o o o D3a Joo O in o O n o o (U 13 S •H X CO 75 vaavno D3a Joo •o in in o in o m o «n in o o <; CTJ 4J 4J Oj ctf S j=: -p ^ -p x: C faO^ • O -H M in S C cd . xi iH r\j Cm nH Cq \ O E C\J e >3 W • O «3 (D 'O Q -H 0 C .H S -H • (XU W • S O 0) E ■Q. ■P H :3 -H a o -P • C B W O -H 4J SH o x; O ^H C cti hO-H O W -H +3 Cm W W J O 0 -H Ctf -P -H O (U fn rH W • Cc; «J a,X3 0 x: S Cti -P O -H :z! • fn C E ■ >5 = 0 -H O -pass •H = O a > rH fn -H •H 0 ■P • hO P< 0 o cti c 'd Cti W O CO ;zS to rH 0 -P >> -H cti x; -H rH O to rH •H to = Oj a cti XI S ,H E Q ctJE ^ -P -P Cd x: cd s p -p ^ c x: o Mx: • e -H w LO C rci • Cm T? rH CM O -H Ph \ S cxj >> a cd w • O Q 0) Ti •H CD C H 6 •H • • Q. U CO in E O cd (U H -H Cm 0 P) :«; s^ us = P cd C S r^ •H cd = d CO S o O cd •H _ 6 a Cm P -H •H P CO ::! O CO 0 Q. 0 rt 0 -H p S O. P rH ::s -H a o -p • c s W O -H P C O ^ O ?H d kj hO-H O CO •H P Cm CO CO J o 0) -H cd P -H O 0 J^ M CO • K cd aX3 0 ^ H cd P O -H it! • !h c: S •>," 0 •H O PCS B •H :: O G > H fn -H •H 0 P • bO tt CD o cd C 73 cd CO O W =! to r-i 0 P >3-H Cd ^ -H H O CO rH •H CO = cd D. cd X) s rH e Q cd = Cm cd * o -=r 0 ' ^ ::i bO •H Ix, 77 Tests were stopped for a particular organism if it died, if no activity was displayed for three consecutive tests, or if the euphausild would not swim up into the test chamber. At the end of 23 days seven of fifteen euphausilds were still alive, but only one was still bloluminescent , so the tests were terminated. (A control group, to determine deterioration of bloluminescent activity caused by daily testing, was not kept.) Five of the eight euphausilds that died did so after a temperature control failure in the laboratory. All of test group one were lost at this time. The results of the parameter evaluations were further analyzed by averaging the activity of each parameter for each euphausild . for : total activity, noon activity, midnight activity and a "before" and "after" moulting (Table II). The moults, found at test time could not be associated with a specific test; therefore the test just prior to finding the moult and. the test just after finding the moult were both evaluated. After the individual averages were made, group averages (Table 3) were accomplished to evaluate the- effect of temperature. The results are plotted on Figures 41-50. These graphs Indicate that two thirds of the euphausilds are more active during the middle of the night than during the middle of the day in responding to artificial stimulation. In the comparison of midnight versus noon activity: eight out of fifteen euphausilds displayed more light output at midnight than at noon; ten of fifteen greater 78 (i's/gwo) V3HV o O 7 ^ o ' O o -a •H •H ta ^ o (A 0) ;^ o CO 0) bO cd fn > cd u < . 1 1 1 I L r c 1 1 1 1 L . 1 1 1 — J r — 1 — r 1 1 1 1 1 1 ilW iJV inn yd St bO o e illM Hd (11 AVQoo -p aViOi 'cs" 0 K> ilW idV ■p a) illAl Hd t_ Ava=«fc •H a -a e C -H •H IViOi mi'] i>iV -P Eh iiw yd Ava^ S < lDHOIM "^ iHDIiM ilW Hd AVG^ iHOIM ilW Hd AVQ f^ J.HDIN * " IViOJ, ilW Hd AVQ fxj iHDIN "* 'IVJ^OJ, ilW Hd ^ a: o Cm (D hO CO !^ > cti ■a •H •H W :3 = cd ^ w P^ (D ::i-H S •H -act: O p) !h O •H fn 0) •H ^VJ,oJ, w 79 (5-2/,Hc) V3HV o O o o ^ 7 iHDIM IViOi Xi~ -P aviOi 03 U iTi'J idV 'O O •H -H •H ?H illM Kd CM Ava ^ m ft ::! = iHDIM oj IViOi ft D :3 -H 0) rH ill'J S^AV ft rH E i^W Hd rH a -rl AVQ ^ :3 X3 Eh iHOIM > s qvioi, •H 'O K C PL. tLT.M J,>3V M ^^ J;1W Vd o AVQ H C\J JLHOIW "^ -3- 'IVJ.OJ, 0) 3 ilW idV bO •H ilW Hd Ph Ava o^ IHDJ.M "* IViOi 80 in T o T (uio) in 1 dWV o ilM Hd AVQoo iHOIM "^ IViOI. J,1M Hd iHDIM nvioi ni'j Hd iHOIN IViOI, ilW idV ilW Hd Ava^ iHDIM IVIOI ilW Hd Ava^ J.HDIM IV 1.0 i j,aw Hd Ava cv^ IHDIiM =«= ' 1V10J- LTA laV J,r[W Hd AvaS! IHDIW IViOl ilW Hd Ava M lllOIM "^^ 7Vi0i bO •H o Amplitude averages for each euphauslld g 1 1 1 -P +j f Xi o •H tH r •H U I cz a QJ I s 1 •H . 1 C Ph 1 on f ^ L (D 1 bO 1 •H Ph 1 1 1 81 in I o T (mo) dWV m 1 o ilM Hd Ava J,HOIM Ava Ln iHDIM * IViOJ, J,1M Hd Ava^ iHOIN =tt i^M vLdV iT>I Hd en Avad iHDIN JjT1'\I Hd CM Ava^ J.HDIM IViOJ, ilW idV ilW Hd Ava M iHDIN ^ ■IViOJ, J,7W Hd Ava H JLHDIW "* ilW idV ilW Hd Ava o^ bO c o Amplitude averages for each euphausiid 1— > Icated plies "after m r- X} s •H 1 1 Eh 0 f -P Bh 1 e< 1 1 KJ • i fan - r for Itln ! hO O 1 1 cu o L- > -p 1 TJ O •H tH •H U 1 m (X J cd 1 0 iH 1 B- M s CZ ri -H x 3 t ( •H ^J > S •H . 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J,HOIM nViOJ, o Ava J,}IDIM tiO •P rH o 6 'O fn :, c £> CO m 0) 0 •H hOrH Oj Q. ?H S QJ •H > rt tH f-J P.S :3 0 cr: ?H PL. 0 ^-^ ON •H 87 (up"} awix in bO c •H •p iH =J o J,™ i.qv E ilW Hd en ■a ^H 0 I •H = iT;j xjv •H 0 •H iiiAi yd CM CO r-\ Ava aV ilW Kd iH ^ < Ava p-l o Cm iHDIN o • IViOi roup ing" vL7W i^V bDp J,1M Jr'd Ava ?H O :3 E iHOIN p IViOi Jh P i^rj idV E o ilW Hd on P U Ava PL, iHDIM O to IViOJ, to (U CD -H hOrH ilL'J iiV fn E J,Tr] J^d CM (D -H Ava Ph cd H IHDIW O as " ^Vl■lOI, 3 o en: ^ Pu J,T;.W i^aV O^ iT.I Hd rH Ava P-, o J.HDIM O LTv r[vj,oi 0) :3 ilW ielV •H J,7W Hd fe Ava iHDIiM qvioi 88 average maximum pulse amplitude at midnight than at noon; and ten of fifteen greater average flash rates at midnight than at noon. The reaction times averaged lower at night than at noon for six of the fifteen. To test the effects of moulting upon blolumlnes- cent activity comparisons were made between the tests prior to moulting and the tests made just after moulting. These two test periods were then compared with total average activity. The average activity indicated by the test just prior to moulting was much greater than that indicated by the test just after moulting. A comparison of "prior to moulting" versus "after moulting" activity shows that eight of eleven euphausllds displayed more light output, eight of eleven displayed greater average maximum amplitude and one the same amplitude and ten of eleven displayed greater average flash rates before than after moulting. The reaction times were lower on the average just prior to moulting than just after for nine of the eleven. (Only eleven of the euphauslid records were used in moulting averages, as there was not enough data for four of the euphausllds . ) Comparisons of tests made just prior to moulting with the total average activity shows that eleven euphausllds displayed more light output, eight of the eleven displayed greater average maximum amplitude and greater average flash rates just prior to moulting than at other times. For the after moulting activity versus total average activity: only four of eleven displayed greater average light output and 89 greater maximum amplitude, and two of eleven euphausilds displayed greater average flash rates. This seems to Indicate greater average blolumlnescent activity (under artificial stimulation) by the euphausilds just prior to moulting and less than average activity after moulting. The group averages show the effect that tempera- ture (assuming that all other variables were actually the same for all groups) has on the measured parameters. The activity at midnight is greater than at noon for all three temperature groups for the parameters of average light output, average maximum amplitude, and average flash rate with one exception. Group one, which was maintained at 12°C, had a greater average light output at the noon test. The same result is true for just prior to moulting tests versus just after moulting tests for the three groups with the same exception. There was greater variation in comparing just prior to moulting tests and just after moulting tests to total average activity. d. Sympathetic Blolumlnescent Response Mauchline (196O) indicated that there may be a sympathetic flash response, i.e. the blolumlnescent response of one organism to the light flash of another. Eight euphausilds were selected, after a postive response to photo.Tlash stimulation, to test this hypothesis and also to determine the presence, if any, of a male-female response such as found in the firefly, Photinus pyralls (McElroy and Sellger, I962)., 90 The euphausllds were placed In individual 100-ml beakers and seven were placed in a circle in a water bath. The eighth euphausiid was photoflash stimulated and placed in the center of the circle. The circle was about twelve inches in diameter and the center euphausiid about six inches from each of the others. The bioluminescence was clearly visible to all other euphausiids due to the swimming patterns of the stimulated euphausiid. The euphausiids were kept in marked containers and each in turn was stimulated and placed in the center. They were allowed to rest for one hour between tests. All euphausiids responded positively to photoflash stimulation, i.e. they bioluminesced, but_ negatively to sympathetic stimulation — at least to visual observation. The observer was given ten minutes to become accustomed to the dark prior to each test. Three males and five female E_^ paciflca with body lengths of from 1^ mm to 19 mm formed the test group. 91 V. DISCUSSION A. AT-SEA RESULTS Useable results in this study were obtained on R/V ACANIA cruises 7^-l4 and 7^-19. Testing of equipment and equipment failure combined with a limited time frame in which to gather information prevented more numerous results . The results from these two cruises are presented. On. ACANIA cruise 7^-1^, bioluminescent activity versus depth tests, using the sensor V7ith the VMT, seemed to indicate that the bioluminescent flash rates of euphausiids may decrease with increases in pressure and/or decreases in temperature. Euphausiids, captured in the upper 100 meters of the water column, and exposed primarily to pressure and temperature changes exhibited significant changes in light output with depth. At depths of 100 meters and below they showed a marked decrease in bioluminescent activity (Figure 11). Kay (1966) determined in laboratory tests that the euphausiid Meganyctiphanes norvegica had increased reaction time to photoflash stimulus with decreasing temperature (two minutes at 7.5°C-12.5°C versus nine minutes at 2.5°C). The temperature change in the water column for our in situ test was from 11°C at the surface to 9.'^°C at 250 meters (Appendix C) . Clarke and Hubbard (1959) recorded bioluminescence to depths of 3750 meters but the effects of pressure changes on bioluminescent activity is unknown. 92 A second In situ test on ACANIA cruise 74-19 also yielded decreased blolumlnescent activity with Increased depth although the blolumlnescent activity of the environment differed somewhat. The environmental bioluminescence on ACANIA cruise 7^-14 decreased much more rapidly with depth. This is depicted in Figure 51. The flash rates for both the test organisms and the surrounding environment are plotted against depth for ACANIA cruise 7^-14 and cruise 7^-19. ACANIA cruise 74-14 tests were made two hours later in the evening than for cruise 74-19. The later tests (cruise 74-14) show a greater concentration of blolumlnescent organisms nearer the surface, probably because a greater percentage of DSL organisms had completed migration in this case. The euphausiid activity depicted is similar in profile to the environmental activity but is dependent upon flash rates of a fixed number of organisms . The decrease in flash rate by the euphausiids could be a defense mechanism (Tett and Kelly, 1973) as the organism is lowered in the water column out of the upper 100 meters v;here caught. Euphausiids were exposed to outside environmental light as a test on ACANIA cruise 74-19 to determine possible effects of environmental light on euphausiid light output . The results of this test Indicated a slight gain in flash rate when exposed to other bioluminescence. Positive response could indicate a sympathetic type reaction, species recognition, or a male-female response (Tett and Kelly, 1973). (Laboratory tests in this study yielded no Intraspecles stimulus — see 93 B. this section.) This activity is also plotted on Figure 51 and depicts the higher activity displayed under these test conditions . Some Investigators (Clarke, I96O; Tett, 1973) suggest that once sufficient background studies have been made a record of the intensity a flash as a function of time or flash "signature" may prove a useful organism identifier. Figure 52a is a bioluminescent background (outside VMT) record of the environment at 100 meters depth recorded at a speed of eight inches per minute. Figure 52b is the bioluminescent record of 10 euphausiids at 50 meters at the same recording speed. Figures 45c and 45d display large E_j_ paclfica (in VMT) flashes at 100 meters, probably individual flashes, at two inches per minute recording speed. Laboratory recordings of individual E_^ paclfica (swimming in a 60 ml beaker) are depicted in Figures 52e and 52f. The laboratory euphausiids had been flash stimulated and sustain activity over a much longer time period. These records Indicate that although an individual order or species of animal may have an individual signature, its extraction from a record of the environmental bioluminescence may prove difficult. This is primarily due to the presence of background light. If this is to be a useable tool the animals will have to be tested individually in situ to obtain a true signature. There is not enough evidence to suggest that euphausiids have a diurnal rhythm of luminescence or that they tend to 94 300 m U -p e -p 0) Q 100 - ■ B^ ■ ©•A ■ ©A ■ ^ ^ A A a ^'O^ ▲

/ V :• : 1 ■ ' ■ 1 ■ ■'!' 1 /l t 1, J, - it M T: 4- J " : :! t • .; ,-" ^ A._ ^ i -iL-jUa >rr-^ lJLM_ i/^N. 1 . .i" X • HjalkliL, .-. ; 1 :::\ !■ .';' ': z- 1 k 1/2 min - ._- 1 . . . ! 1 1- :: ' . — . — , . 1 ?'" ^^ ' J ) — 1 ■ : Figure 5.2. Blolumlnescent records of environment and Individual E^ pacifica. a. Environment at 100 m depth b. 10 euphausiids at 50 m in VMT c,d. Large euphausiid flashes at 100 m in VMT e,f .Laboratory recordings of light output of individual E. pacifica. •96 bioluminesence more at dusk and at dawn (Mauchllne and Fisher, 1969). The answers to these questions are Important in trying to determine the function of euphauslld biolumlnescence and what natural levels of luminescence to expect In migrating or stationary layers of euphausllds. (This reasoning could also be applied to other blolumlnescent DSL organisms.) If biolumlnescent activity is greatest during migration it may be used for maintaining spacing between individuals during migration, species identification during migration and keeping continuity in the migrating layer, or for separation between layers of different animals to prevent mixing of the layers (Tett and Kelly, 1973). The photomultlplier sensor used with the VMT may provide answers to some of the above. There are no field tests of in situ activity of bioluml- nescence with which to compare our results and the number of tests run require guarded conclusions. B.. LABORATORY RESULTS The results of the laboratory tests from this study are presented here. The reader is again reminded (Chapter II) that there are many unknowns in this type of testing and in trying to relate the results to the behavior of the test organism in its natural environment. 1. Results of Spontaneous Tests Mauchllne (i960) in tests for spontaneous bioluml- nescence on the euphauslld, Meganyctlphanes norvegica, and Tett (1972) in similar tests on M. norvegica and 97 Thysanoessa raschll determined that this type of activity is rare and unpredictable in the laboratory. However most of these animals die within a few days after capture, demonstrating inability to live in the artificial laboratory environment. E. pacifica has demonstrated a much longer life expectancy in the lab - on the order of months (Lasker and Theilacker, 1965; Komaki, I966). Five tests were run in the present study to try to relate spontaneous activity to time of day. If a pattern were determinable it might yield Information on the functions of biolurainescence . Examination of the records of the activity of the five groups did not reveal any Identifiable pattern (Figure 15). The euphauslids were active for periods which varied from 100% of the time in.a ten-hour test group (Group 3) to 20% of the time in a 3^ hour test group (Group 5). A feature of all of the spontaneous activity was the low amplitude (0.1 mv, see Figure I8) response indicating weak flashes or possibly a soft glow type bloluminescence exhibited on several occasions by euphauslids while under visual observation. The initial stimulus, caused by handling the euphauslids when moving them to the test equipment, lasted 15 to 20 minutes and was of high (up to 2.5 mv) amplitude. 2. Results of Bioluminescent Activity Versus Ambient Light The ambient light levels for E^ pacifica in its own environment is about 10 yV//cm (Kampa and Boden, 195^^). Tests to determine the effects of different light levels on 98 the blolumlnescent activity of laboratory euphausllds were run on a large (25) group of E^ pacifica. A relatively bright light did seem to cause a faster loss of blolumlnescent activity. The other light intensity categories; "dark", "dim", and alternating "dark and dim" were Inconclusive, A concurrent moulting test was also Inconclusive. An important result of this study was the development of new laboratory test equipment and methods for further study. 3. Results of Blolumlnescent Activity Versus Temperature, Moulting and Time of Day. If the euphausiids use bioluminescence for some specific function, i.e. feeding in the near surface water, they might exhibit more activity in the laboratory under stimulation, during that time period they would normally have been feeding. The euphausiids were tested for a relative blolumlnescent activity at noon and at midnight. It was suspected that at these two periods corresponding to feeding at the surface or resting at depth the greatest difference in activity would occur. Another possible period of great activity was that of migration. This had the disadvantage that if tested against a non migration time period, equal time periods between tests would be difficult to set up and therefore might bias the results. The results of the tests Indicate that two-thirds of the euphausiids shov/ed a greater response to stimulation at the midnight test than at the noon test. V/hen the individual results were averaged within their respective temperature 99 groups, the preference for night over day was more pronounced. Therefore, excluding effects of pressure and other parameters and time periods not tested, euphausllds appear to demonstrate greater use of the blolumlnescent function during the night, near-surface period of their diurnal cycle. The results of blolumlnescent activity during moulting periods indicated that the average activity In the tests prior to moulting was greater than the average activity in the tests after moulting by a large percentage . This would be the expected result, as other organisms normally have a quiescent period following moulting. This same relation was evident in comparing the time of moulting with average activity and in the group comparison tests. C. GENERAL DISCUSSION AND FUTURE USE OF VMT The photomultlpller light detector is a valuable tool in the study of underwater light but has been limited in some aspects of its use (Boden and Kampa, 1964; Mauchllne and Fisher, 1969; Tett and Kelly, 1973). Work in marine bioluml- nescence studies has lost impetus in recent years (Tett and Kelly, 1973), and it is felt that this may be due, in part, to the slow progress in underwater light detector methodology. The VMT utilized with the underwater light sensor introduces a new approach to in situ studies. If laboratory tests are run in conjunction with in situ studies, further Insight into the functions of marine blolumlnescence v;ill undoubtedly be gained. 100 •Three of the major problems encountered in studies of marine blolumlnescence using the photomultlpller light detector are solved In part by use of the VMT with the sensor. A specific organism can be studied in situ; but it is physi- cally restrained to a certain volume of that environment. The distance and angle from the organism to the light detector are maintained within a given restricted range. The artificial stimulus due to the vertical oscillation of the sensor in the water column is decreased to some degree. The physical stirring and overpressure caused by the sensor's movement through the v;ater are absent, but other factors, such as cable vibration carried to the VMT, may still cause some stimulation. Euphausllds could be easily maintained and monitored through a 2H hour period in the VMT. A suitable high frequency fathometer could be used to monlter DSL depths and keep the VMT within the DSL. The blolumlnescent activity could then be, monitored through a 24 hour cycle. The apparatus could also be used to test for blolumlnescent changes in periods of euphausild swarming, for near-surface activity (feeding), for at-depth activity, and for changes with time, light and other parameters. The VMT can be used to hold organisms for testing within their own environment with little change to that environment, or it may be used to partially control certain of the physical and chemical elements of the environment. Light may be fully or partially masked out. The small access holes can be enlarged and 101 increased in number (or a flow-through system included) to allow oxygen, pH, salinity and nutrient changes to occur more rapidly within the tube . The tube ( or a shorter version) could be filled with water from selected depths to vary the parameters to other levels of the water column 102 VI. CONCLUSIONS 1. An underwater test chamber to retain organisms for testing within their own environment was designed and constructed. This chamber, called a vertical migration tube (VMT)j allows partial control of the physical and ecological factors of the organism's environment during testing. 2. The use of the vertical migration tube with a photomultlplier sensor m.ay prove to be a valuable tool for in situ mesopelagic studies. Initial tests of the unit demonstrated the feasibility of its use In the marine environment . 3. Three major problems associated with studies of marine biolumlnescence using an underwater photomultlplier light detector are resolved in part by the use of the VMT with this sensor. With it a sp.ecific (known) organism can be studied in situ; the distance and angle from the organism to the sensor can be controlled; and the artificial stimulus due to the vertical oscillation of the sensor in the water column is minimized. 4. Euphausllds taken in the upper 100 meters of the water column at night decrease their bloluminescent flash rates when lowered In the water column and exposed primarily to pressure and temperature changes. This could imply that a defensive mechanism or a feeding mechanism is at work. 10 3 5. There may be an Increase In euphauslid blolumlnescent flash rates when stimulated by other blolumlnescent organisms. This could be an intraspecies response and be either a sympathetic type reaction, species recognition, or a male- female type response. An Interspecies response may be responsible also and used to provide species exclusion. 6. The laboratory test for 12- to 2'1-hour activity rhythms were inconclusive. Low amplitude bioluminescence was a feature of all the spontaneous activity, and this type of bioluminescence may be the "normal" light output of the euphauslid. Bright light or mechanical stirring (artificial laboratory stimulus) elicited much higher amplitude activity. 7. Laboratory test equipment was designed and built to allow a quantitative measurement to be made of the light output of a small marine organism. This equipment when used with a photomultlpller light detector and recorder allows measurements of total light output, amplitude and flash frequency to be made. 8. The use of a flash "signature" (flash amplitude as a function of time) to identify a species or an order of marine organisms, although possible, may prove difficult due to the presence of other background light. 9. A "bright" light laboratory environment seemed to cause a faster loss of blolumlnescent activity in a group of test euphausiids. The ambient light level of their -H 2 normal environment is on the order of 10 yW/cm . It was lOM concluded that a "dim" light environment ( 10 -^ yW/cm ) was best to maintain the euphauslids in the laboratory. 10. Laboratory tests indicated a greater bioluminescent response to a standard flash stimulus during midnight tests as opposed to noon tests. This may indicate that the euphausiid's "normal" use of bioluminescence occurs at that time. This might be associated with feeding or another surface related function. 11. Laboaratory tests of bioluminescent activity Indicated greater than average response to a photoflash stimulus just prior to moulting and less than average response just after moulting. This would appear to be a normal physiological condition. 105 TABLE I Record of the measured parameters of the dally bloluminescence of each E. pacifica TOTAL MAX LIGHT OUTPUT FLASH FLASH REACTION DATE AREA AMP FREQ Tir^; MOULT Euphausla pacifica #1: 13 Feb n 8.0 7.8 19 0.6 M 13 Feb m 1.0 3.0 3 1.9 16 Feb n 9.4 7.0 10 1.1 16 Feb m 3.8 3.0 7 0.3 17 Feb n 0.8 0.5 1 1.4 17 Feb ra 0.6 0.3 1 0.3 18 Feb n 0.4 1.9 3 1.7 18 Feb m 5.6 4.5 3 2.0 M Euphausla pacifica §2 13 Feb n 1.3 4.8 6 0.8 13 Feb m 5.9 6.7 16 1.3 Euphi ausia pacifica #3: 13 Feb n 45.2 13.4 32 0.3 13 Feb m 40.9 13.4 35 0.6 16 Feb n 33.8 13.4 32 0.5 16 Feb m 32.2 13.4 39 0.5 17 Feb n 62.2 13.4 19 0.5 17 Feb m 13.3 12.7 13 0.8 18 Feb n 16.1 5.5 5 1.0 18 Feb m 1.8 0.5 1 1.1 M 106 TABLE I (continued) TOTAL MAX LIGHT OUTPUT ELASH FLASH REACTION DATE AFEA AMP FREQ ausia pacifica #4: TIME MOULT Euphj 13 Feb n 1.2 2.3 2 0.9 13 Feb m 1.8 2.3 4 1.2 16 Feb n 1.7 0.5 2 5.3 16 Feb m 32.5 13.4 50 1.6 17 Feb n 14.2 13.4 17 0.6 17 Feb m 0.7 2.5 2 2.4 18 Feb n 0.4 0.1 1 1.0 M 18 Feb m 8.1 6.3 4 1.2 Euphausia pacifica #5 13 Feb n 0.6 0.6 1 1.1 13 Feb m 1.5 1.5 1 2.2 16 Feb n 4.5 3.5 13 2.1 16 Feb m 3.3 4.9 _. 5 0.3 17 Feb n 4.2 4.6 4 1.6 17 Feb m 0.2 1.1 2 3.6 18 Feb n 0.7 1.1 2 3.6 18 Feb m 0.5 0.8 2 5.0 M 107 TABLE I (continued) TOTAL MAX LIGHT OUTPUT FLASH FL'^SH REACTION DATE AREA AMP ausla paclfi FREQ ca #6: TIME MOULT Euphi 13 Feb n 1.1 1.1 2 1.8 13 Feb m ^.3 3.7 7 2.1 16 Feb n 3.3 1.3 9 3.7 M 16 Feb m 30.9 13.4 18 0.3 17 Feb n 10.9 7.5 19 1.3 17 Feb m 2.l\ 3.8 6 1.8 18 Feb n 1.3 2.0 4 3.0 18 Feb m 15.7 13.4 18 1.1 22 Feb m 0.1 0.1 0 0.7 M 23 Feb n 0.1 0.1 0 0.9 2i} Feb n 1.2 1.1 7 3.2 25 Feb n 4.6 6.6 12 2.4 26 Feb n 0.5 0.7 1 2.9 27 Feb n 6.8 6.1 23 1.6 2 8 Feb m 0.4 1.1 1 1.5 M 2 Mar n 0.2 1.3 3 2.4 3 Mar n 2.0 1.6 3 1.9 3 Mar m 0.0 0.0 0. 6.0 5 Mar n 1.1 1.7 3 3.8 6 Mar n 0.0 0.0 0 6.3 M 108 TABLE I (continued) TOTAL MAX LIGHT OUTPUT FLASH FLASH FEACriON DATE AREA AMP J^'KEQ ausia paclfica #7: TIME MOULT Euphc 13 Feb n 1.2 1.0 3 1.2 13 Feb m 7.5 3.0 5 1.3 16 Feb n 0.1 0.1 0 4.7 16 Feb m 0.1 1.0 -1 0.9 17 Feb n 0.3 0.8 H 3.0 17 Feb m 0.5 1.1 6 2.1 18 Feb n 0.0 0.0 0 6.0 18 Feb m 0.0 0.0 0 6.0 M 109 TABLE I (continued) TOTAL MAX LIGHT OUi'PUT FLASH FLASH REACTION DATE AREA A^P ia paciflca FREQ #8: TIME MOULT Euphaus 13 Feb n 17.2 13.4 41 0.7 M 13 Feb m 23.1 13.4 50 0.6 16 Feb n 32.3 13.4 41 1.3 16 Feb m 7.5 5.0 34 0.8 17 Feb n H.l 5.5 53 1.3 17 Feb m 7.2 9.1 11 1.2 18 Feb n 10.2 13.4 17 0.9 18 Feb m 9.7 12.6 24 1.0 M 22' Feb m 0.1 0.2 0 6.0 23 Feb n 0.5 1.1 7 4.1 24 Feb n 5.1 10.1 29 2.1 25 Feb n 2.5 6.3 9 1.6 M 26 Feb n 10.2 12.2 18 1-3 27 Feb n 1.8 5.1 14 2.0 2 8 Feb m 0.0 0.0 0 6.0 2 Mar n 0.7 2.1 5 2.3 3 Mar n 0.6 1.3 2 2.7 3 Mar m 0.0 0.0 0 6.0 M 110 TABLE I (continued) DATE TOTAL TJGHT OUTPUT AREA MAX FLASH AMP ia paclflca FLASH l^'KEQ #9: REACTION TIME MOULT Euphaus 13 Feb n 2.7 1.0 1 3.5 13 Feb m 50.7 13.4 8 0.3 16 Feb n 15.3 11.6 6 0.6 16 Feb m 43.7 13.4 10 0.3 17 Feb n 0.7 3.5 4 2,0 17 Feb m 10.7 12.5 10 0.6 18 Feb n 2.5 9.2 10 1.0 18 Feb m 0.4 1.4 2 1.8 22 Feb 23 Feb m n 1.1 0.5 4.2 1.0 4 6 0.3 1.8 M (19th) 24 Feb n 12. i| 13.4 23 0.4 25 Feb n 3.0 0.7 1 3.9 M 26 Feb n 3.0 3.4 6 3.0 27 Feb n 3.4 3.6 - 14 1.8 2 8 Feb m 0.6 3.5 4 1.4 2 Mar n 0.2 0.6 1 3.4 3 Mar n 1.3 3.2 2 1.6 3 Mar m 3.5 0.4 18 1.6 5 Mar n 0.0 0.0 0 6.3 M 111 TABLE I (continued) TOTAL MAX LIGffl? OUTPUT FLASH FLASH REACTION DATE AREA AMP la pacifica l-'HEQ #10: TIME MOULT Euphaus 13 Feb n 3.6 3.8 23 1.0 M 13 Feb m 7.9 3.8 26 1.0 16 Feb n 11.8 9.1 22 2.1 16 Feb m 58.9 13.4 58 0.3 17 Feb n 1.7 1.8 3 2.8 17 Feb m 26.2 12.5 19 1.8 Euphausla pacifica #11: 16 Feb m 15.7 13.4 32 1.3 17 Feb n 11.7 12.4 21 0.6 17 Feb m 3.3 5.1 14 1.2 18 Feb n 5.2 5.6 19 0.7 18 Feb m 2.3 6.7 16 1.2 22 Feb m 1.6 4.5 3 1.9 M 23 Feb n 0.6 1-2 - 0 4.2 24 Feb n 2.2 5.1 37 2.0 25 Feb n 1.9 13.4 11 0.9 26 Feb n 0.5 0.2 1 1.3 27 Feb n 7.3 10.2 19 1.1 2 8 Feb m 0.1 0.5 1 2.4 2 Mar n 0.0 0.0 0 6.0 3 Mar n 1.3 2.6 3 2.0 3 Mar m 0.6 1.3 2 1.5 5 Mar n 0.3 1.1 2 4.0 6 Mar n 0.4 0.8 1 3.2 112 TABLE I (continued) TOTAL MAX T,TGHT OU'i'PUT FLASH FLASH REACTION DATE AREA AMP la pacifica FREQ #12: TIME MOULT Euphaus 16 Feb m 32.4 13.4 45 0.5 17 Feb n 8.9 11.5 40 1.3 17 Feb m 21.3 12.3 86 1.8 18 Feb n 21.7 13.4 20 1.2 18 Feb m 40.9 13.4 .39 0.4 22 Feb m 7.7 11.0 13 4.2 M 2 3 Feb n 2.9 4.7 9 0.9 24 Feb n 2.3 3.5 6 3.4 25 Feb n 0.6 2.3 2 2.4 26 Feb n 3.7 6.6 13 1.5 27 Feb n 4.6 2.0 11 ' 0.5 28 Feb m 9.3 11.5 16 0.3 2 Mar n 0.7 1.8 3 0.8 3 Mar n 13.8 13.4 ' ■ 22 0.4 3 Mar m 4.3 7.2 4 2.1 5 Mar n 3.8 4.8 6 1.5 6 Mar n 1.1 1.4 1 3.5 M 113 TABLE I (continued) TOTAL MAX LIQfT OUTPUT FLASH FLASH REACTION DATE AREA AMP la paclflca FREQ #13: TIME MDULT Euphaus 16 Feb m 2l\.6 "12.6 17 1.5 17 Feb n 6.0 2.4 3 1.3 17 Feb m 1.3 1-5 0 3.8 18 Feb n 2.9 2.1 0 4.7 18 Feb m 10.8 7.6 18 4.0 22 Feb m 0.7 0.0 0 4.6 M 23 Feb n 0.0 0.5 0 6.0 2k Feb n 0.0 0.1 8 6.0 114 TABLE I (continued) TOTAL MAX LIGHT OUTPUT FLASH FLASH REACTION DATE AREA AMP la pacifica i-'HEQ #14: TIME MDULT Euphaus 16 Feb m 33.9 13.4 ^3 0.4 17 Feb n 20.0 10.1 50 0.5 17 Feb m 13.7 5.7 60 1.4 18 Feb n 3.1 4.4 14 1.4 18 Feb m 18.5 8.6 45 1.3 22 Feb m 27.3 13.4 57 1.5 M 2 3 Feb n 3.6 13.4 30 1.2 24 Feb n 5.6 13.4 16 1.2 25 Feb n 22.7 13.4 31 1.1 26 Feb n 13.5 13.4 26 0.8 27 Feb n 6.6 13.4 12 1.3 28 Feb m 9.0 6.1 7 1.4 M 2 Mar n 2.9 3.7 23 1.8 3 Mar n 8.6 13.4 -■ 19 1.4 3 Mar m 26.6 13.4 21 0.9 5 Mar n 7.5 11.9 15 1.5 6 Mar n 7.4 13.4 22 1.6 Euphausla pacifica #15: 16 Feb m 49.9 13.4 43 1.2 17 Feb n 21.6 12.2 25 0.4 17 Feb m 2.2 2.2 4 3.8 18 Feb n 0.8 1.1 1 6.0 18 Feb m no test M 115 TABLE II Averages of Each of the Measured Parameters of the Dally Bilumlnescent Activity of E. Paclflca BEFORE AFTER E. paclflca AVG NIGHT DAY MOULT MOULT #1 Total Light Output (Area) 3.7 2.75- 4.65' . 0.4 6.8 Max . Amp . 3.5 2.7 4.3 1.9 3.75 Plash Freq. 5.8 3.5 8.15 3.3 11.05 Reaction Time 1.16 1.12 1.2 1.7 1.3 #2 Total Light Output (Area) 3.6 5.9 1.3 Max. Amp. 5-75 6.7 4.8 Flash Freq. 10.8 l6.1 5.5 Reaction Time 1.05 1.3 0.8 #3 Total Light Output (Area) 29.4 22.1 36.8 32.2 60.0 Max. Amp. 10.7 10.0 11.4 13.4 13.4 Flash Freq. 22.0 22.0 22.0 39.0 19.0 Reaction Time 0.66 0.75 0.57 0.5 0.5 #4 Total Light Output (Area) 7-57 10. 87 4.37 0.7 0.4 Max. Amp. 5.1 6.12 4.07 2.5 0.1 Flash Freq. 11.04 15.O 5.57 2.0 1.3 Reaction Time 1.78 1.6 1.95 2.4 1.0 116 TABLE II (continued) BEFORE AFTER E. paclflca AVG NIGHT DAY MOULT MOULT #5 Total Light Output (Area) 1.93 1.37 2.49 3.3 2.38 Max . Amp . 2.26 2.07 2.45 4.9 2.6 Flash Freq. 3.53 2.15 4.9 4.5 2.55 Reaction Time 2.^ii 2.78 2.1 0.3 1.35 #6 Total Light Output (Area) 4.35 7.69 2.57 4.07 1.23 Max. Amp. 3.33 5.07 2.14 3.83 0.8 Flash Freq. 6.62 7.14 5.63 11.0 3.43 Reaction Time 2.53 1.93 2.14 2.5 3.73 #7 Total Light Output (Area) 1.62 2.7 0.53 Max. Amp. 1^7 1.67 0.6 3 Flash Freq. 2.37 2.5 2.27 Reaction Time 2.2 1.43 2.97 #8 Total Light Output (Area) 7.81 7.93 10.83 5.3 2.03 Max. Amp. 7.31 6.72 8.65 8.27 6.3 Flash Freq. 20.88 19.83 28.83 I6.0 11.0 Reaction Time 2.11 2.6 1.72 1.9 2.87 117 TABLE II (continued) BEFORE AFTER E. paclflca AVG NIGHT DAY MOULT MOULT #9 Total Light Output (Area) 8.66 15.81 3.13 7.95 3.23 Max . Amp . 6.0 8.11 ^.3 10.9 0.35 Flash Freq. 7.22 8.0 4.29 20.5 0.5 Reaction Time 1.62 0.9 3.05 1.0 4.5 #10 Total Light Output (Area) 18.35 31.0 5.7 Max. Amp. 7.4 9.93 4.9 Plash Freq. 25.17 34.33 16.0 Reaction Time 1.5 1.03 1.97 #11 Total Light Output (Area) 4.29 3.93 2.85 2.3 0.6 Max. Amp. 5^71 5.25 6.15 6.7 1.2 Flash Freq. 13.91 11.31 l6.5 l6.0 0.0 Reaction Time 1.67 1.58 1.67 1.2 4.2 #12 Total Light Output (Area) 14.17 19.32 9.03 40.9 7.7 Max. Amp. 10.29 12.5 8.08 13.4 11.0 Flash Freq. 20.08 22.17 l8.0 39.0 13.0 Reaction Time 1.44 1.6 1.28 0.4 4.2 118 TABLE II (continued) BEFORE AFTER E. paclfica AVG NIGHT DAY MOULT MOULT #13 Total Light Output (Area) 5.79 9.35 2.22 10.08 0.7 Max . Amp . 3.34 5.4 1.27 7.6 0.0 Flash Freq. 5.75 6.25 2.75 18.0 0.0 Reaction Time 3.79 3.47 4.5 4.0 4.6 #14 Total Light Output (Area) 14.6 21.4 7.92 12.5. l8.1 Max. Amp. 10.1 10.1 11.3 11.0 9.75 Flash Freq. 30.3 38.8 21.8 28.5 32.0 Reaction Time l.l8 1.2 1.1? 1.3 1.45 #15 Total Light Output (Area) l8.4 25.6 11.2 Max. Amp. 7.22 7.8 6.65 Flash Freq. 20.7 23.5 13.0 Reaction Time 2.85 2.5 3.2 119 TABLE III Averages of Each Measured Parameter for Each Temperature Group of E. Paclflca GROUP 1 BEFORE AFTER (12°C) AVG NIGHT DAY MOULT MOULT Total Light Output (Area) 9.24 8.58 9.92 9.15 17.9 Max. Amp. 5.46 5.52 5.40 5.42 4.96 Flash Freq. 10.7 11.71 9.22 12.2 8.47 Reaction Time 1.42 1.51 1.52 1.22 1.04 GROUP 2 (alt. temp. 7°C-12°C) Total Light Output (Area) 8.8I I3.O 4.59 4.42 2.17 Max. Amp. 5.21 6.31 3.70 7.76 2.48 Flash Freq. 12.88 14.36 11.4 15. 83 4.98 Reaction Time 1.91 1.58 2.42 1.8 3.7 GROUP (7°C) 3 Total Light Output (Area) Max. J ^mp . Flash Freq. React: Lon Time 11.46 15.9 6.64 16.64 6.78 6.31 8.21 6.7 9.67 5.49 18.16 20.4 14.4 25.37 10.75 1.64 2.07 2.38 1.72 3.61 120 APPENDIX A ACANIA Cruise No. P g -Eg •H O +J ^^ -p 00 rH ::! M ^8 (D "to Eh g •H +:> rci -p OQ on o O -H O -P -p CO 8 rH a o ^ ^ p 0 'to g •H -P Cti -P 00 o o o s p^ o o o -p w 0) Eh O (U O C/3 O 0 U H H -P H 0) -P tn on 3fcJ O > ■P O 'O 73-67 73-71 73-78 73-80 73-82 73-87 73-88 73-90 73-95 73-96 73-101 7^-1 74-6 7^-14 74-16 74-19 74-22 74-30 73 74 10 11 12 01 02 03 1 X 14 X 6 X 17 X 25 X 4 X 9 X 17 X 12 X 13 X 6 X 9 X 24 X 7 X 8 X 14 X 22 X 11 X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X *Fluorometer studies v;ere conducted on early cruises while the photomultiplier detector and VI4T were being designed and constructed. A hose was lowered (to depths of 75 meters) and water punped up to a surface fluorometer to measure bioluminescence . 121 APPENDIX B Q O 00 Ph H H W O H pL, o to S Species of Euphausiid at CalCOFI Station Number Three m t^ en 00 ^ o> 00 C-- t>- ^T t- m rH t^ cr» a^ >- cr> t— cr> rH rH 0>i r-\ -^ ON •% rH r-i h- H 1^ •\ *» #\ J rH •» X3 f^ 0) 0) >J ?H -p fci 0) x> X) J^ cd «\ CO 0 C ^ u :3 0 0 0 0) d 0) cri < W 0 a M <^ Pq s XXX XX XXX Euphausla paclflca Nyctlphanes simplex XXX X Thysanoessa spinifera X Thysanoessa gregaria X X Nematoscelis dlf f icllis XX XX Stylocheiron longicorne Stylocheiron maximum Identification of species was accomplished using a key in Mauchline and Fisher (I969). The entire net haul (100 meter vertical haul) v/as examined only if very few (10-20) specajnens were present. An aliquot part of the haul was examined if the number of euphausiids present was estimated at more than 20 as follovjs: one half if estimated from 20 to 40; one third if estimated from 40 to 100; and one tenth if estimated more than 100 euphausiids present. 122 APPENDIX C SAMPLE XBT TAKEN AT CalCOFI Station 3 1 r 10 15 TEMPERATURE (°C) 20 25 SHIP ACANIA -CRUISE T'^-l^ LAT CalCOFI 3 LONG TIME 18^0L DA/MO/YF I 1/2/lk 123 APPENDIX D w \i 3000 5000 MH mmm Spectral-sensitivity characteristic of phototube having S-li response to radiant flux fron a tungsten source at 2870° K (from RCA data sheet number 92CK-6652R3), 12^ APPENDIX E Calibration curves for 1P21 photomultlplier tube T ILLUiMCATION AT DETECTOR (foot-candles x 14. U3) 125 LIST OF REFERENCES 1. Barham, E.G., The Ecology of Sonic Scattering Layers In the Monterey Bay Area, California, Ph.D. Thesis, Stanford University, 1956. 2. Beebe, W., Half Mile Down, 3^4 p. , Harcourt Brace and Co., New York, 1934. 3. Boden, B.P. and Kampa, E.M., "Records of biolumlnescence in the ocean," Pacific Science, p. 229-235, April, 1957. 4. Boden, B.P., Kampa, E.M. and Snodgrass, J. M. , "Measurements of spontaneous biolumlnescence in the sea." Nature , Vol. 208 (5015) pg. 1078-1080. Dec. 11, I965. 5. Boden, B.P. and Kampa, E.M., "Planktonic biolumlnescence," Oceanography and Marine Biology Annual Review, Vol. 2, p. 341-371, 1964. 6. Breslau, L.R. and Edgerton, H.E., "The luminescence camera," Journal of the Biological Photographic Association, Vol. 26, No. 2, p. 49-5», May, 1958. 7. Brown, R.H., Preliminary Analysis of the Detection of Objects by Biolumlnescence, NRL Report 7065, June 2, 1970 . 8. Clarke, G.L. and Backus, R.H., "Measurements of light penetration in relation to vertical migration and records of luminescence of deep sea animals," Deep-Sea Research, Vol. 4, p. 1-14, 1956. 9. Clarke, G.L. and Breslau, L.R., "Studies of luminescent flashing in Phosphorescent Bay, Puerto Rico, and in the Gulf of Naples using a portable bathyphotometer," Bulletin Institute oceanographlque , Monaco, No. 1171, I960 , 32 p. 10. Clarke, G.L. and Hubbard, C.J., "Quantitative records of the luminescent flashing of oceanic animals at great depths," Limnology and Oceanography, Vol. 4, No. 2, p. 163-180, April, 1959. 11. Clarke, G.L. and Kelly, M.G., "Measurements of Diurnal Changes in Biolumlnescence from the Sea Surface to 2,000 meters using a new Photometric Device," Limnology and Oceanography, Vol 10, p. 54-66, I965. i26 12. Clarke, G.L. and Wertheim, G.K., "Measurements of Illumi- nation at great depths and at night in the Atlantic Ocean by means of a new bathyphotometer," Deep-Sea Research, Vol. 3, p. 189-205, 1956. 13. Hardy, A.C. and R.H. Kay, "Experimental studies on plankton luminescence," Journal of Marine Biological Association of United Kingdom, Vol. 44, p. 435-^b^, 1964 . 14. Hardy, A.C, and Paton, W.N., "Experiments on the Vertical Migrations of Planktonic Animals," Journal of Marine Biological Association of United Kingdom, Vol. 26, p. 467-526, 1947. 15. Home, R.A,, Marine Chemistry, John Wiley and Sons, New York, 568 p. , 1969. 16. Kampa, E.M. and Boden, B.P., "Submarine illumination and the twilight movements of sonic scattering layer. Nature, London, vol. 17^, p. 869-87O, 195^. 17. Komaki, Y., "Technical notes on keeping euphauslid live in the laboratory, with a review on experimental studies on euphausiids ," Information Bulletin on Planktology, Japan, Vol. 13, p. 95-105, 1966. 18. Lasker, R. and Theilacker, G.H., "Maintenance of euphauslid shrimps in the laboratory. Limnology and Oceanography, Vol. 10, p. 287-288, I965. 19. Mauchline, J., "The biology of the euphauslid crustacean, Meganyctiphanes norvegica (M. Sars)," Proceedings of the . Royal Society, Edinburgh, Biology, Vol. 67, p. 141-179, I960. 20. Mauchline, J. and Fisher, L.R., "Biology of the euphausiids," Advances in Marine Biology, vol. 7, p. 1-454, 1969. 21. McElroy, V/.D., and Seliger, H.H., Biological luminescence Readings from Scientific American. W.H. Freeman and Co., San Francisco and London. 196«, p. 128-140, 1962. 22. Nicol, J.A.C., "Observations on luminescence in pelagic animals. Journal of the Marine Biological Association, United Kingdom, Vol. 37, P- 705-752, 1958. 23. Neshyba, S., "Pulsed light stimulation of marine bioluminescence in situ," Limnology and Oceanography, Vol. 12, p. 222-235, 1967. 127 2k. Nlcol, J.A.C., The Biology of Marine Animals, Pitman and Sons, London^ 2nd edition, 699 pp. 1967. 25. Rudykov, J. A. C. , "Procedures for studying the luminescence of the sea," Oceanology, Vol. 7, No. 4, p. 569-577, 1968. 26. Tett, P.B., "An annual cycle of flash Induced lumines- cence in the euphausiid Thysanoessa raschil , " Marine Biology, Vol. 12, p. 207-218, 1972. 27. Tett, P.B. and Kelly, M.G., "Marine Biolumlnescence ," Oceanography and Marine Biology Annual Review, Vol. 11, p. 89-173, 1973. 128 INITIAL DISTRIBUTION LIST No. Copies 1. Defense Documentation Center 2 Ca!meron Station Alexandria, Virginia 2231^1 2. Library, Code 0212 2 Naval Postgraduate School Monterey, California 939^0 3. Department of Oceanography, Code 58 3 Naval Postgraduate School Monterey, California 939^0 4. Oceanographer of the Navy 1 Hoffman II 200 Stovall Street Alexandria, Virginia 22332 5. Office of Naval Research 1 Code ^80 Arlington, Virginia 22217 6. Dr, Robert E. Stevenson 1 Scientific Liaison Office Scripps Institution of Oceanography La Jolla, California 92037 7. Library, Code 3330 1 Naval Oceanographic Office Washington, D.C. 20373 8. SIO Library 1 University of California, San Diego P.O. Box 2367 La Jolla, California 92037 9. Department of Oceanography Library 1 University of V/ashington Seattle, VJashington 98IO5 10. Department of Oceanography Library 1 Oregon State University Corvallls, Oregon 97331 11. LCDR Calvin Dunlap 6 Oceanography Department Naval Postgraduate School Monterey, California 939^0 129 12. Dr. Stevens P. Tucker Department of Oceanography Naval Postgraduate School Monterey, California 93940 13. Dr. Peyton Cunningham Department of Physics and Chemistry Naval Postgraduate School Monterey, California 93940 14. Library, Hopkins Marine Station Hopkins Marine Station Pacific Grove, California 93950 15. Library Moss Landing Marine Laboratories Moss Landing, California 95039 16. Mr. Larry Ott Naval Air Development Center Johnsville, Warminster, Pennsylvania 18974 17. Mr. John Shannon Naval Air Development Center Johnsville, Warminster, Pennsylvania 18974 18. Dr. Hasong Pak Department of Oceanography Oregon State University Corvallis, Oregon 97331 19. Dr. Kendell Carder Marine Science Institute University of South Florida St. Petersburg, Florida 337 01 20. LCDR Andrew J. Compton Naval Undersea Center San Diego, California 92132 21. Dr. F. G. Walton Smith Rosenstiel School of Marine and Atmospheric Science Rickenbacker Causev/ay Miami, Florida 33149 130 SECURITY CLASSIFICATION OF THIS PAGE (»hmn D.l. Enlorod) REPORT DOCUMENTATION PAGE t. REPORT NUMBER 2. GOVT ACCESSION NO 4. TITLE ("and Subl/t/eJ A Study of the Blolumlnescence of a Deep Scattering Layer Organism (Euphausla paclfica) in Monterey Bay, California 7. AUTHORfs; Andrew Jerome Comptom 9. PERFORMING ORGANIZATION NAME AND ADDRESS Naval Postgraduate School Monterey, California 939^0 READ INSTRUCTIONS BEFORE COMPLETING FORM 3- RECIPIENT'S CATALOG NUMBER 5. TYPE OF REPORT & PERIOD COVERED Master's Thesis; _jviarch^ 19 7^ 6. PERFORMING ORG. REPORT NUMBER 8. CONTRACT OR GRANT NUMBERr»; 10. PROGR«,M ELEMENT. PROJECT. TASK AREA ft WORK UNIT NUMBERS II. CONTROLLING OFFICE NAME AND ADDRESS Naval Postgraduate School Monterey, California 939^0 12. REPORT DATE March 197^ 13. NUMBER OF PAGES 132 1«. MONITORING AGENCY NAME « ADDRESSf// dy//oren( from Controlling Olllce) Naval Postgraduate School Monterey, California 939^0 IS. SECURITY CLASS, fof Ihia tiport) Unclassified ISa. DECLASSIFI CATION/ DOWN GRADING SCHEDULE 16. DISTRIBUTION STATEMENT (ol Ihia Report) Approved for public release; distribution unlimited. 17. DISTRIBUTION STATEMENT (ol the abilracl entered In Block 30, II dlllerer\l from Report) 18. SUPPLEMENTARY NOTES 19. KEY WORDS (Continue on reverse aide If neceeaary and Identity by block number) Blolumlnescence Euphausla paclfica Deep Scattering Layer 20. ABSTRACT (Continue on reveree tide It neceamary and Identity by block number) A vertical migration tube (VMT) v;as designed and constructed as an instrument to be used with a photom.ultiplier light detector to make in situ mesopelagic studies of deep scattering layer vertical migration organisms. Initial tests of the unit demonstrated the feasibility of its use in the marine environment Three major problems of marine biolum.inescent studies using an underwater photomultlplier detector are resolved in part by the DD 1 J AN "73 1473 EDITION OF I NOV 6S IS OBSOLETE (Page 1) S/N 0102-OM- 6601 I 131 SECURITY CLASSIFICATION OF THIS PAGE (When Data Bntered) CtCUHITY CLASSIFICATION OF THIS PAGEfH'hen D«(« Enttrmd) (20. ABSTRACT continued) use of the VMT with this sensor. Mesopelaglc euphauslld crustaceans captured in the upper 100 meters of the water column at night decreased their bioluminescent flash rates V7hen lowered in the water column and exposed primarily to pressure and temperature changes. There may be an increase in euphausiid bioluminescent flash rates when stimulated by other bioluminescent organisms. Laboratory test equipment and laboratory methods were developed to permit quantitative measurements of euphausiid bioluminescent output. Laboratory tests of Euphausia pacifica indicated a greater bioluminescent response to a standard flash stimulus during midnight tests as opposed to noon tests. Laboratory tests of bioluminescent activity during periods of moulting indicated greater than average response to a photoflash stimulus just prior to moulting and less than average response just after moulting. DD Form 1473 (BACK) 1 Jan 73 S/N 01U2-014-6G01 , security classification of this PACEf«7i»n d«(» em. «<<; 132 Thesis 15G4S8 C6578 c.l Compton A study of the bio- luminescence of a deep scattering layer organ- ism (Euphausia pacificaj in Monterey Bay, Cali-J fornia. J 19 4 jAMT-r 2 385 1 1 1 1 1 M "r T u U 0 O KAY 87 3 2 0 2 8 SEP 87 3 00 56 Thesis C6578 .1 1.5L4S8 Compton A study of the bio- luminescence of a deep scattering layer organ- i sm (Euphausia pacifica) in Monterey Bay, Cali- fornia. thesC6578 A Study of the bioluminescence of a deep 3 2768 002 09292 6 DUDLEY KNOX LIBRARY ^