University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Nebraska Game and Parks Commission -- Staff Nebraska Game and Parks Commission Research Publications

1984 A Guide to Time of Death in Selected Wildlife Species D.W. Oates Nebraska Game and Parks Commission

J.L. Coggin Nebraska Game and Parks Commission

F.E. Hartman Nebraska Game and Parks Commission

G.I. Hoilien Nebraska Game and Parks Commission

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Oates, D.W.; Coggin, J.L.; Hartman, F.E.; and Hoilien, G.I., "A Guide to Time of Death in Selected Wildlife Species" (1984). Nebraska Game and Parks Commission -- Staff Research Publications. 64. http://digitalcommons.unl.edu/nebgamestaff/64

This Article is brought to you for free and open access by the Nebraska Game and Parks Commission at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Nebraska Game and Parks Commission -- Staff Research Publications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. A GUIDE TO TIME OF DEATH ­ IN SELEGED WILDLIFE SPECIES

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' & NEBRASKA GAME & PARKS COMMISSION ------'IJ Eugene T. Mahoney, Director ____----J A GUIDE TO TIME OF DEATH IN SELECTED WILDLIFE SPECIES by D. W. Oates, J. L. Coggin, F. E. Hartman and G.!. Hoilien

Nebrasl"la Technical Series No. 14 Nebrasl"la Game & Parl"ls Commission Lincoln, Nebrasl"la 1984

A Contribution of Federal Aid in Wildlife Restoration, Project W-57-R ACKNOWLEDGEMENTS Appreciation is extended to all conservation officers, wildlife biologists and wildlife area supervisors who collected and made collections possible. Special thanks to Dr. John Gill of Maine and the Journal of Wildlife Management; Jim Pex and Ken Meneely of Oregon and the Journal of Forensic Science: John Olson, Larry Rhinehart and fellow conserva­ tion officers from Indiana; Dennis Martin, Ken Sexton, John Pound and Dennis Mullins from Virginia; Dr. Alan Woolf and John Will from Illinois; George Gage, Dean Knauer, Bob Barratt and Jim Kienzler from Iowa; Ken Kerr and Patrick Karns from Minnesota; Tom Morrow, Fred Glover, Richard Denny, Harlan Riffel, Bob Mosley, and Cathy Hoilien from Colorado; and William Shope and Dr . John Kreider of Pennsylvania. Editorial assistance by Liz Huff, Dr. Curtis Twedt, Bill Baxter and Ken Johnson; sample collection and preparation by Carol Jochum; data tabulation by Joe Gabig and Kit Hams; typing by Beverly Read, Margo Ems, Ruth Wusk; typesetting by Jan Bouc; art work by Chris Mercer, Paula Day, and Julie Beeson, and printing by Leland Busch and Rosemary Upton were deeply appreciated. Previously un­ published data supplied by several states was invaluable for this study.

5 TABLE OF CONTENTS

Page

ACKNOWLEDGEMENTS ...... 5

LIST OF FIGURES...... 7

LIST OF TABLES ...... 8

LIST OF COLOR PLATES ...... 9

ABSTRACT ...... 10 INTRODUCTION ...... 11 TEMPERATURE ...... 12

ELECTRICAL STIMULUS ...... 12

RIGOR MORTIS...... ; ...... 13

PHYSICAL CHANGES IN THE EYE ...... 13

CHEMICAL CHANGES IN THE EYE...... 14

POTASSIUM ...... 14

GLUCOSE...... 14

DEER...... 17

TEMPERATURE ...... 17

ELECTRICAL STIMULUS ...... 19

RIGOR MORTIS...... 30

PHYSICAL CHANGES IN THE EYE ...... 32

CHEMICAL CHANGES IN THE EYE ...... 32

GLUCOSE...... 34 POTASSIUM ...... 34

WATERFOWL...... 37 TEMPERATURE (Mallard duck) ...... 37

RIGOR MORTIS (Mallard duck) ...... 37

EYE APPEARANCE ...... 40

TEMPERATURE (Geese)...... 41 ELECTRICAL STIMULUS (Geese)...... 43

PHYSICAL CHANGES IN THE EYE (Geese) ...... 45

CHEMICAL CHANGES IN THE EYE ...... 45

POTASSIUM ...... 45

PHEASANTS...... 49

TEMPERA TURE ...... 49

ELECTRICAL STIMULUS ...... 49 . . .

PHYSICAL CONDITION OF THE EYE ...... 55

CHEMICAL CHANGE ...... , ...... 55

6 Table of Contents, Cont. Page

POTASSIUM ...... 55 COTTONTAIL RABBIT...... 63

TEMPERATURE ...... • ...... 63 ELECTRICAL STIMULUS ...... 63 BLACK BEAR ...... 67 TEMPERATURE ...... 67 ELK ...... 68 TEMPERATURE ...... 68 RACCOON ...... 69 TEMPERATURE ...... 69 ELECTRICAL STIMULUS ...... 69 LITERATURE CITED ...... 71

LIST OF FIGURES Figure Page

1 Equipment used in conducting electrical stimulus tests...... 13 2 Pictorial representation of the eye showing locations for the aqueous and vitreous humor 14 3 Diagrams depicting the location for collecting vitreous humor from a deer ...... , 14 4 Diagram depicting the "best" location for collecting vitreous humor from a bird ...... " 15 5 Live weight of white.tailed deer based on field·dressedweight of Nebraska deer ...... 18 6 Diagram depicts insertion of two types of thermometers.Sensor should be placed in center of muscle mass in thigh...... 18 7 Diagram depicts insertion depth for naso.phanyngeal thermometer and measurement of pupil diameter...... 19 8 Schematics of a deer carcass with electrode positioning marked to denote possible test location ...... 27 9 Post·mortem aging curves as influencedby ambient temperature. Body temperature data were taken from dry mallards whose average weights were approximately 2.6 lbs. (courtesy Morrow and Glover, 1967 .. " 38 10 Post·mortem aging curves as influencedby ambient temperature. Body temperature data taken from dry mallards whose average weights were approximately 2.2 lbs. (courtesy Morrow and Glover, 1967) ...... 38 11 Post-mortem aging curves as influencedby ambient temperature. Body temperature data were taken from mallards placed in water of ambient temperature for three minutes immediately following death and whose average weights were approximately 2.5 lbs. (courtesy Morrow and Glover, 1967) ...... 39 12 Post-mortem aging curves as influencedby ambient temperature. Body temperature data were taken from mallards placed in water of ambient temperature for three minutes immediately following death and whose average weights were approximately 2.2 1bs. (courtesy of Morrow & Glover, 1967)...... 39 13 Worcester's data showing thoracic change in temperature with time in cackling geese ...... , 41 14 Worcester's data showing thoracic change in temperature with time in snow geese ...... 42 15 Cloacal temperature change in Canada geese. Symbols represent birds weighing from 6.7 to 121bs...... 42 16 Locations on a goose where electrical stimulus was checked...... 43 17 Potassium levels in the vitreous humor of goose eyes with respect to time ...... , 45 18 Average breast temperature of male pheasants at 400F ambient temperature in various situations ...... " 50 19 Thigh temperatures of individual male pheasants at 500F ambient temperature ...... " 50 20 Body temperature of adult pheasants taken cloacally at an ambient temperature of 40°F ...... , 51 21 Average cooling rate at selected locations on male pheasants at 40°F ...... 51 22 Average cooling rate at selected locations on male pheasant at 500F ...... , 52 23 The rate of decline in thigh temperatures at three ambient temperatures ...... 52 24 Male pheasant data collected at Graterford prison hunt, November 197 3. Ambient temperature was 32 420F ...... 56

7 25 Potassium levels in the vitreous humor Iowa pheasant eyes ...... 57 26 Potassium levels in the vitreous humor of Nebraska pheasant eyes...... 57

27 Post-mortem cooling of cottontail rabbits. Condition 1 - Single rabbit, dry unskinned, uncleaned ...... 64

28 Post-mortem cooling of cottontail rabbits. Condition 2 - paired rabbits, dry unskinned, uncleaned and laying together ...... 64

29 Post-mortem cooling of cottontail rabbits. Condition 3 - single rabbit , unskinned, uncleaned and laying on a wet surface ...... 65 30 Locations on a rabbit where electrical stimulus was checked ...... 65

31 Diagram depicting typical muscle response at four locations in cottontail rabbit ...... , 65

32 Post-mortem temperatures in the mouths of black bear with respect to time in hours ...... " 67 33 Post-mortem temperatures in the ears of black bear with respect to time in hours ...... 68

34 Post-mortem muscle temperature in elk with respect to time in hours ...... 69 35 Post-mortem cooling curves for day raccoons ranging in weight from 7 .5 to 15 pounds and at ambient temperatures varying from 40 to 600F ...... 70

36 Locations on a raccoon where electrical stimulus tests may be conducted ...... , 69

LIST OF TABLES

Table Page

1 Field-dressed weights(carcass less entrails, heart, liver and lungs) of white-tailed deer, based upon heart girth . .. 17 2 Time of death estimates of white-tailed deer based on weight and post-mortem changes in thigh and nasal temperatures ...... 19 3 Electrical stimulus results are presented for six different time intervals and seven different muscle groups. Response was recorded as being very good, good, fair, poor and none ...... 28

4 Muscle response to electrical stimulus in Illinois white-tailed deer ...... , 28 5 Rigor mortis observations in white-tailed deer, based on data from Indiana and Gill and O'Meara (1965) ...... 29 6 Time afterde ath in hours when either no or full rigor was found in 70-99% of the Maine (Gill and O'Meara,

1965) and Indiana deer ...... , 30 7 Rigor mortis observation in deer from Nebraska (based upon rigor or no rigor present)...... 30

8 Time after death in hours when either rigor or no rigor was observed 70 to 99% of Nebraska deer ...... " 30 9 Expectations for the majority of yearling adult white-tailed deer that have been dead for a selected number of hours...... 31 10 Distribution of adult white-tailed deer pupil diameters (mm) by post-mortem intervals in deer examined in

Maine (Gill and O'Meara, 1965), Illinois (Woolf, 1983), Virginia and Nebraska ...... , 32 11 Distribution of fawn white-tailed deer pupil diameters (mm) by post-mortem intervals in deer from Maine (Gill and O'Meara, 1965), Illinois (Woolf, 1983), Virginia, Indiana and Nebraska...... 32

12 Multiple linear regression analysis for nasal temperature and vitreous humor potassium and glucose in moose . . . 33 13 Scattergram of potassium data in the vitreous humor of white-tailed deer eyes versus the post-mortem interval in hours ...... 33

14 Prediction of time interval (hours) when glucose levels are known in blacktail deer ...... 34 15 Observation frequency of certain post-mortem characteristics of mallard eyes; arrow indicates the general progression of change ...... 40 16 Snow goose electrical stimulus data showing reactions of six muscle groups with respect to time ...... 44

17 Canada goose electrical stimulus data showing reactions of six muscle groups with respect to time ...... " 44 18 Cooling rates of male pheasant thighs and breasts based upon 95% confidence limits and an ambient temperature of 400F ...... 53 19 Cooling rates of male pheasant thighs and breasts based upon 95% confidence limits and an ambient temperature of 500F ...... " 54 20 Cooling rates of male pheasant thighs and breasts based upon 95% confidencelimits and an ambient temperature of 600P ...... 55 21 Post-mortem reactions to electrical stimulus in cottontail rabbits ...... 66

8 LIST OF COLOR PLATES

Plate Page

I DEER EYES ...... 35

II GOOSE EYES ...... 47 III PHEASANT EYES ...... 59

N PHEASANT EYES ...... 60

V PHEASANT EYES ...... 61

VI PHEASANT EYES ...... 62

9 ABSTRACT The objectives of this study were to monitor post­ mortem changes in the eyes of slected species, as well as other physical changes. In order to obtain a more suitable manuscript, people from other states were contacted. Their data, along with some published data, was included in this manuscript. We were, therefore, able to cover post-mortem changes in several species. For deer, data was collected on tempera­ ture, electrical stimulus, rigor mortis, pupil diameter, pictor­ ial changes, and chemical changes in the potassium and glucose levels within the eye. Waterfowl data included temperatures, rigor mortis, physical changes in the eye, electrical stimulus and changes in potassium levels in the vitreous humor of the eye. In pheasants, changes were primarily with tempera­ ture and physical and chemical changes in the eye. In pheasants, changes were primarily with tempera­ ture and physical and chemical changes in the eye. Cottontail rabbit data dealt with both temperature and electrical stimulus. Data on black bear, elk and raccoon dealt predominately with temperature. Much of the data (especially deer) was supplied by cooperative hunters, and the remainder predominately by wildlife biologists. Much of the data is presented in tables. Hopefully it is in a form usable by conservation officers and field biologists. Most of the information on time of death (TOD) requires an "on the spot" evaluation of existing data. This means that field personnel will most likely have to make the final decision on TOD.

10 INTRODUCTION The ability to estimate time of death (TaD) in wildlife species is a skill greatly desired by law enforcement per­ sonnel (Beattie and Giles, 1979). It can be critical to a conservation officer whether a deer was killed during the early hours of open season or the night before, since it is not uncommon to have a deer checked in that was killed a day before the season opening. Other concerns may be the illegal taking of game by spotlighting or the instance where daily bag limits aren't identical to possession limits and the daily bag limit has been exceeded. These and many other examples could be cited to demonstrate the need for information to aid in TOD estima­ tions. Coe (1974) and Van Den Oever (19 76) provided a review of human post-mortem literature which includes wildlife research. Research into estimating time of death in wildlife has continued sporatically since 1968. This research has been carried out independently by several state fish and game agencies, the U.S. Fish and Wildlife Service and at times by university students. Unfortunately , most of this work appears never to have been formally published. Since infor­ mation on the subject of TOD in wildlife species is rather limited and widespread, this guide was compiled in an attempt to aid field personnel by consolidating some of the existing data. The physical characteristics examined were carcass temperature, response to electrical stimulus, rigor mortis and physical changes in the eye. Chemical changes were also monitored. TaD estimates normally should not be based on obser­ vation of one parameter. However, examination of a combin­ ation of several parameters coupled with good "old horse sense" should allow one to estimate TOD with reason­ able accuracy. These estimations most often are based on an "on-the-spot" one-time examination of the carcass and therefore require a field rather than laboratory examination. For the sake of convenience, information will be pre­ sented firstby test.parameter and then QV �pecies.

11 TEMPERATURE instead of mercury and should be checked for calibration After death, body temperature approaches ambient before use. temperature through loss of heat by radiation, convection (2) A flexible 6·foot tape measure to measure heart girth and conduction. Monitoring of these temperature changes of deer. as an indicator of time since death is not a recent innova­ (3) Notebook and pencil for recording data. tion. In fact, Rainy attempted to explain the cooling phe­ (4) Cloth for cleaning thermometers, etc. nomena in humans as early as 1869. Brown and Marshall *For peace of mind and insurance, carry at least one (1974), presented a review of existing literature on human spare thermometer. cooling curves and attempts to derive mathematical descrip­ tiems of these curves. ELECTRICAL STIMULUS Cooling rates depend primarily upon: (1) initial body Response to electrical stimulus is chemically related to temperature: air temperature ; (3) body surface area the availability of adenosine triphosphate (ATP) in muscles. and weight and(2) (4) handling of the carcass after death. Of After death ATP breaks down and response to electrical these parameters several are relatively constant. Initial body stimulus gradually decreases until no response is observed. temperatures are normally similar for a given species. Devia­ The physiological variables which affect rigor can also affect tion from the norm, however, can occur in diseased or response to electrical stimulus. Rested, healthy, well nour­ stressed individuals. ished individuals that die quietly should respond to electrical Body surface area remains constant unless an animal is stimulus longer than an individual which undergoes a severe field dressed. From that point on, the surface area is essen­ death struggle or is fatigued or stressed prior to death. Ina­ tially constant; however, a considerable variation in this bility to respond to electrical stimulus may be hastened by parameter can occur in individuals within a species. quick death under extreme exertion, tension or brain injury. The weather is among the more variable parameters. Biological texts (e.g. Gamong, 1981 ; Keeton, 1969; Morrow , Air temperatures may vary over a substantial range during a 1968) can be consulted if additional information on this given day. When co:r.sidering this parameter, wind chill phenomenon is desired. and humidity are very important criteria. Both of these The value of this test lies in its ability to allow TaD esti­ factors can increase the cooling rate. mates for the first few hours (4 or less) after death which are The most variable parameter involves handling of the not always possible with a thermometer. Muscles give a carcass after death. One can logically assume that fielddr ess­ strong response to electrical stimulus immediately following ing, skinning, transportation in the back or on top of an open death. With the passage of time, this reaction decreases in vehicle will hasten cooling. However, one would expect the certain muscle groups in varying degrees. This variation may opposite affect if the carcass was placed by the heater or allow one to relate TOD to the degree or presence of placed with several other carcasses in an enclosed area. response in selected muscle groups. These and many other factors may have to be assessed when Muscle response to electrical stimulus was observed making a TOD estimate based upon carcass temperature. when a low frequency electrical impulse! (supplied by a Normally, several measurements made over a period of time vehicle) was applied to muscles of deer, pheasants, rabbits, in a more controlled environment will improve one's ability ducks, geese and raccoons. These responses were recorded as to make an accurate estimate. being: (1 ) Very Good - gross muscular contractions - response Methods and Materials normally observed even in muscle groups that were not Rectal temperature measurements were normally taken, touched by the electrodes. with approximately 3·inch insertion, in rabbits, ducks, geese, Good strong muscular contractions obsened in pheasants and raccoons. Two temperatures were measured in muscles(2) touched by electrodes. deer: (1) center of muscle mass in hind leg; (2) naso· (3) Fair slight tWitching corresponding to tIle elcc�ri pharyngeal cavity of the head. Deer weights can be estimated cal impulses. by heart girth measurements, but actual weights should be (4) Poor no twitching observed, just slight nlLlsc�e taken when possible. tightening which may be easily observed on an exposed

Some thoracic, breast and thigh temperatures were taken muscle or may appear to be a slight change in hair or fea:h":'f on waterfowl and pheasants. Consult tables or text to make positions in an undressed specimen (may be difficult to sure you are looking at the correct data. view under windy field conditions). (5) None no visible muscle contraction.

Materials (6) Extremely Good 2 - single gross muscular response Several thermometers * one at least 12 inches long to often accompanied an appropriate verbal release and dropping fit in(I) the nasal cavity of deer. Thermometers should be of the electrodes (normally observed when electrodes either either the meat type with a dial or filled with red alcohol touched tester or tester touched ungrounded or wet testee).

lSeveral vehicles were utilized during this study. All of these vehicles and many pGst-1975 vehicles are equipped with a high energy ignition system. Consequently , our test voltages were probably exceeding 10,000 volts. Private communications with auto mechanics instructors and service personnel would indicate that little or no damage should occur to a vehicle if time spent in testing was not extensive . Problems may arise in vehicles equipped with computer systems. Testing may cause "check engine" light to come on. Local car dealers can be consulted fo r details on your particular vehicle if this occurs. Correction of this problem was reportedly not difficult.

2 This reaction may be somewhat alleviated if small animals are placed on a rubber or plastic floormat prior to testing.

12 RIGOR MORTIS Rigor mortis is a post-mortem state of rigidity which developes in muscle tissues when their supply of A TP (adenosine triphosphate) and phosphocreatine has been depleted. This depletion takes place via a chemical reaction and is dependent upon both physiological parameters and ambient temperature. The presence of rigor is normally evaluated by the manipulation of selected joints. A fairly reliable rigor sequence has been derived for humans (Morrow, B 1968) and deer (Gill and O'Meara, 1965). This sequence has been observed to be reversed 24 to 48 hours afterdea th in deer (Morrow, 1968 and Gill and O'Meara, 1965) and from 30 -80 hours in humans (Van Den Oever, 1976). This loss of rigor has been credited to the putrefaction process. Several laboratory devices impractical for field use have been devised to measure rigor (Marsh, 1952; Bate-Smith and Bendall, 1956, Briskey, et aI., 1962). Field people, how­ ever, will have to flex joints and rely on visual estimates such as full rigor, partial rigorand no rigor. Gill and O'Meara (1965) cautioned that TOD estimates based upon rigor can be confused by the effects of carcass handling, freezing temperatures and gunshot wounds.

Equipment (1) Notebook and pencil or pen to record data.

PHYSICAL CHANGES IN THE EYE A Attach to hot spark plug wire FollOWing death several interesting phenomena occur which can aid time of death estimates. Among these changes B Attach to ground on engine are loss of eye clarity, luminosity and shape, color changes in the pupil, and constriction of the pupil. At death, the cornea and eye fluids are very transparent and the surface Figure 1. Equipment used in conducting electrical stimulus tests. of the eye is tight and smooth. The transparency and lumin­ osity decrease as the intraocular fluids become cloudy. Eye shape changes as internal fluid pressure diminishes; putre­ Hoilien (1976) described the equipment and procedures faction occurs and the liquid volume is reduced. Gayett for performing electrical stimulus tests in the field. (1900) attached very little diagnostic significance to these The equipment consisted of a 10-foot piece #14 two changes, which appear to be related to temperature, humid­ conductor double-insulated wire, equipped with four well ity and air currents (Lyle et aI., 1959), Color changes have insulated alligator clamps (Figure 1). been observed in white-tailed deer and mallard duck eyes by A spark plug wire is disconnected from a spark plug or Gill and O'Meara (1965) and Morrow and Glover (1967a). the distributor cap and one of the clamps can be attached to Following complete pupil dilation shortly after death, the hot wire inside the plug insulator (an alligator clamp with gradual post-mortem contraction resulting from rigor of the jaws of at least I-inch length or a short piece of metal rod or iris muscles has been ob served in humans (Wurdemann, tubing the same size as the top of the spark plug may facil­ 1920; Saram, 1957) and deer (Gill and O'Meara, 1965). itate this attachment). The second clamp was attached to the Color photos were taken of white-tailed deer, ring­ engine and served as ground. Two 1 �-inch syringe needles necked pheasant and snow geese in an attempt to portray were attached to the alligator clamps in the testing end to some of these changes. facilitate penetration through hair and feathers to the skin (not through). The vehicle can now be started and con­ Methods and Materials nections checked by putting the needles � to �-inch apart. A Physical changes in the eye were recorded photogra­ spark should jump between the two needles. The two needles phically for pheasants, geese and deer and are presented in can then be placed at selected locations on the animal to the back of this manual. For deer, observations were also monitor muscle response . made on both the luminosity of the eye and pupil con­ striction. Materials (1) One, 10-foot piece of #14 double insulated wire Materials equipped with well insulated alligator clamps on at least (1) Camera, if one wants a permanent visible record one end. for court purposes. Best results were obtained with a 200 (2) Vehicle with high energy ignition system. mm or 500 mm telephoto lens for deer to illustrate lumines­ (3) Notebook and pencil or pen for recording data. cence and pupil closure.

13 (2) Dividers for measuring pupil diameters. Preferably Glucose a pair you can operate with one hand. As with potassium, glucose research has been primarily a (3) Metric ruler for pupil measurements. concern in human pathology (Sturner and Gantner, 1964). (4) Flashlight to aid in pupil and luminescence mea­ In humans the glycolysis process was considered by Coe surements. (1972) but completed witrun 3� to 7 hours (Hamilton­ (5) Notebook and pencil for recording data. Patterson and Johnson, 1940). In dogs, Schoning and Strafus (1980) found glucose levels in the vitreous humor to be

CHEMICAL CHANGES IN THE VITREOUS related to time and temperature in post-mortem intervals of HUMOR OF THE EYE 3 hours or less. Pex et al. (1983) found a linear relationship Vitreous humor is a somewhat viscous liquid (especially that was relatively independent of temperature and appar­ in deer) found in the posterior chamber of the eye (Figure ently suitable for determining time of death in blacktail 2). The anterior chamber contains the aqueous humor (a less deer. This technique was associated with thigh temperature volatile liquid). Following death, potassium levels increase in measurements and found suitable for time of death estimates the vitreous humor due to the release of potassium as the for the first7 hours. retinal cells break down (Jaffe , 1962; Hughes, 1965). Con­ If the eyes have been damaged, and the fluids appear versely glucose levels decrease due to glycolysis (Hamilton­ bloody or have retinal material floating in them; the tests Patterson and Johnson, 1940). for glucose or potassium should not be used.

Potassium Levels in the Vitreous Humor Vitreous humor was aspirated from the eye into a plastic syringe (5cc syringe with an 18 gauge, I-inch needle for deer and a 1 cc syringe with a 21 gauge , I-inch needle for pheas­ ants and geese). Prior to needle insertion, the syringe plunger was drawn back about � centimeter. After insertion the plunger was depressed, thus alleviating some of the plugged needle problems. Penetration of the lens and dislodgement of retinal material should be avoided. In deer, the vitreous humor may be gelatinous and very viscous. The needle would occasionally become clogged by fibrous material. Our best success came when we placed the needle tip into the central area of the posterior chamber (Figures 2 and 3) and then applied suction and external pressure on the eyeball with finger or thumb while rotating and moving the needle tip.

Figure 2. Pictorial representation of eye showing locations for the aqueous (A) and vitreous (B) humor.

Potassium Studies on humans in the early 1960's demonstrated a linear relationship between potassium levels in the vitreous humor and time of death for a period up to 125 hours. Results were reportedly independent of temperature and could be estimated to within ten hours or less (Adelson, et a1 ., 1963; Sturner and Gantner, 1964). Considerably different confidence limits have been placed upon post­ mortem interval estimates based upon vitreous potassium levels (Van Den Oever, 1976). Recently, ambient temper­ ature was found to have an effect on potassium levels (Schoning and Strafuss, 1980). Johnson et al. (1980) found a logrithmic correlation in mule deer in Utah rather than the linear relationship observed in humans. We observed a linear relationship for white-tailed deer, ring-necked pheasants and geese. The above information along with Woolf and Gre­ million - Smith's (1983) review should provide some insight Figure 3. Diagrams depicting the location for collecting into the current usefulness of this technique. vitreous humor from a deer.

14 The technique must be modified when taking the vit­ Glucose Levels in the Aqueous Humor of Blacktail Deer reous humor from pheasants and geese. First puncture the With 1 ml syringe, collect 0.5 ml of aqueous humor from membrane protecting the membrane protecting the aqueous anterior chamber of eye (see Figure 2). Transfer sample to humor with a needle and then blot the aqueous humor with a a vacutainer tube containing sodium fluoride preservative, towel. Since the retina coats the inside of the scherotic then refrigerate if possible until sending to lab. Centrifuge (white) membrane one must penetrate the eye at an angle sample, add ul of clean supernatant to 1 ml of 6% 0- almost perpendicular to it (Figure 4). The eye is penetrated toluidine, incubSO ate for 10 minutes at 1000C, cool to room at the outer part of the colored portion. Withdraw the plunger temperature and read in a spectrophotometer at 630 mm. about � inch, insert needle � to � inch, miss the lens, Measurements below 5 mg/100 mg may be limited by low depress plunger and draw out about � ml of the clear vitre­ optical density readings (pex et a1. 1983). ous humor. Extend pressure on the outer surface of the eye­ ball may facilitate matters. Field Equipment

Syringes - 1 ml (l)(2) Syringe needles - I-inch, 21 gauge (3) Vacutainer tubes containing sodium fluoride preservative (4) Notebook and pencil for recording data.

Lab Equipment (1) Centrifuge (2) Visible range spectrophotometer

Glucose Levels In The Vitreous Humor For Moose (Karns and Kerr, 1984a) and White-tailed Deer (Karns and Kerr, 1984b) Using a 1 �-inch, 18 gauge needle collect about 3 ml of sample (2 - 4 mI). Red blood cells should be centrifuged immediately after collection, for they will decrease glycogen levels; avoid when possible. Glycolysis was found to be quite temperature-dependent and should be carried out between ambient temperatures of 230 to 410 F to Figure 4. Diagram depicting the "best" location for col­ +50 C). Stress can also cause elevated glucose levels(-SO in the lecting vitreous humor from a bird. blood and eye, so if the deer is wounded or chased before being shot it should not be included in this data. Homogen­ ous, fiHered samples were used for analysis (Schleicher and Volumes for samples varied, but normally at least 0.5 Schull 0.2 micrometer, 25mm disposible; Gelman and milliliters could be collected from each eye. Samples were Millipore make similar filters). As the fluid begins to come then centrifuged at 2,000 rpm for 5 minutes; the clear through the filter, add hypodermic needle and insert in 5 supernatant was returned to the syringe and refrigerated ml plain vacutainer tube. The clear, colorless vitreous humor at 40 C until analysis. was analysed immediately or frozen until analysis. Analysis Duplicates were run on each sample. Twenty-five ul was carried out using the Orthotoluidine Diagnostic kit of alignots were diluted to 25 ml in volumetric flask. Potas­ Signa Chemical Company. Glucose values were found to sium levels were measured with an atomic absorption spec­ decrease logarithmically for approximately 24 hours. trophotometer. In case of disagreement, an additional set of duplicates was run. Field Equipment (1) 18 gauge, 1 �-inch needle Field Equipment (2) 5 ml sterile disposible syringe (1) Syringes (3) 25 mm, 0.2 micrometer filter andho lder and 5 ml for deer hypodermic needle 1 ml for geese and pheasants (4) 5 ml plain vacutainer (2) Syringe needles notebook and pencil for recording data 18 gauge, I-inch for deer (5) 21 gauge, I-inch for geese and pheasants Laboratory Equipment (3) Centrifuge or millipore filteringsystem (1) Visable range spectrophotometer (4) Notebook and pencil for recording data. (2) Ortho toluidine Diagnostic Kit No. 635 or possibly Phenol-methyl-salicylate (PMS) Lab Equipment method by Hycel, Boeringer-Mannheim (1) Flame photometer or atomic absorption spectro­ Company might work because of the extremely photometer. acid reaction conditions. (2) Assorted glassware including appropriate pipets and volumetric flasks.

15 DEER

Time of death has been estimated by several techniques. Virginia Temperature measurements and response to electrical stim­ Data for developing TOD charts in Virginia was taken ulus may be the most reliable of these techniques. Monitor­ on the Quantico Marine Base at Quantico, Virginia, by ing of chemical and physical changes in the eye and the rigor Virginia State Game Wardens and Biologists. mortis sequence can also provide valuable information. The photographs depicting luminosity and lens closure were taken on the Gathright Wildlife Management Area in DEER SAMPLE COLLECTION western Virginia by Research Biologist, Dennis Martin.

Nebraska TEMPERATURE Mule and white-tailed deer samples were collected at The normal body temperature of a live deer is approxi­ selected deer check stations across the state during the 1980, mately 1020 F; however, temperatures as low as 1000F 1981 and 1982 rifle seasons. Most of the samples were have been observed (Gill and O'Meara, 1965). acquired during special muzzleloader seasons at the DeSoto Stressed or diseased deer may have temperatures higher National Wildlife Refuge near Blair and Gifford Educational than 1020F. Agreeing with Gill and O'Meara (1965), temper­ Game and Woodlands near Bellevue. Time of death was atures as high as 1070F were observed in Nebraska and supplied by cooperating hunters. Indiana. To employ existing temperature data, one should know lliinois approximately both the ambient temperature for the first This data was furnished by Dr. Alan Woolf of the Coop­ 12 hours after death and the weight of the deer. For normal erative Wildlife Research Laboratory at Southern Illinois University-Carbondale and John Will of the Illinois Con­ servation Commission. Much of their data was collected at Heart NE NE NE VA VA PA WI the Crab Orchard National Wildlife Refuge in southern Girth N=22 N=25 N=59 N=144 N=384 N=13 Illinois during the 1981 firearm season. Time of death was Adult primarily based on information supplied by cooperating (in.) Fawn Doe Buck Fawn Deer Deer Deer hunters. 25 Maine 26 Supplied through the cooperation of the Journal of 27 Wildlife Management and Dr. John Gill, data resulted from a 28 herd reduction program at Acadia National Park in 1961 29 56 64 68 64 and 1962. Deer were shot by National Park Service per­ 30 82 59 70 73 69 sonnel, so actual time of death is known. Deer were handled 31 86 86 63 76 79 74 under simulated fieldcon ditions. 32 90 92 66 85 80

Indiana Data came from John Olson, Larry Rhinehart and fellow conservation officers. Data was collected not only at Crane 36 Naval Weapons Support Area but also at Kingsbury, Atter­ 37 111 129 11 bury and Patoka Fish and Wildlife areas during the 1982 rifle 38 115 138 119 125 season. Actual time of death was supplied by cooperating 39 119 148 1 25 134 hunters. 40 124 159 131 144 41 128 169.··.. 1.37 Oregon 42 132 180 143 Most of the samples were collected at the McDonald 43 137 194 149 Forest Check Station. Blacktail data was supplied by Jim 44 141 208 · · 1 55 Pex and Ken Meneely of the Oregon State Police Crime 45 224 161 Detection Laboratory in Eugene, the Journal of Forensic 46 240 232 Science , and Dan Carleson of the Oregon Department of Fish 47 250 and Wildlife . Except for Carleson's data the actual time of 48 272 death was supplied by the hunters.

Minnesota Table 1. Field-dressed weights (carcass less entrails, heart Samples were collected primarily in the St. Croix area liver and lungs (lbs.)) of white-tailed deer, based upon during special firearms seasons in 1980. Data on glucose and heart girth. Data is from Nebraska, Virginia (Smart, et aI. potassium in vitreous humor will be published in 1984 by 1973) Pennsylvania (Cowan, et aI. 1968) and Wisconsin Patrick Karns and Ken Kerr of the Minnesota Department of (Trail Sign Products, P.O. Box 365, Germantown, WI Natural Resources in Grand Rapids. 53022).

17 fall days in Maine, Gill and O'Meara (1965) found that one I --- - 32 (1-- - c � could obtain a reasonable estimate of daily temperatures by ° 1------31 - --- 1- ,-- I-i------taking one temperature reading at dawn and another at 30 f- - -- I-- -�--- f- t - ,. �*- r I I 3 p.m. These readings should make it possible to place 1---- ,- f--- - 29 r- -- samples in a fairly specific temperature category if approxi­ 28II l- i- -- i-i--- - I--- : mate weight is known. The field-dressed weight of the deer l ! __ 27 : 1------f- ····_JtJ l I not including heart, liver and lungs, can be estimated by 26 a I � -- f-- I - measuring the circumference of the chest just behind the 250 '/ - I front legs (heart girth). 24 0 Y -t -- -- - I I / For Nebraska deer, we found a relationship similar to 23 a ���,- - = .-- I-- - =i� - that found in Pennsylvania (Cowan. et a1. 1968) for adult 22 0 / --r- v- i bucks. Virginia (Smart, et a1. 1973) examined over five­ 21o -- - � -L ! hundred deer; 144 were fawns and the remainder were 20 r� r� / i r o HI -� I-- 190 i �= unsegregated adults. Their fawn data was relatively close to :r: I / : I i I ! S2 18 01--- - I Nebraska's. Table 1 is an estimate of weight, the gene pool, UJ / I I i - i I $:17v / I available food, sex and life style are very important in UJ i V I ! ! > determining a deers actual weight. For best estimate , use if 16 0 I- i I :::i ! I 150 Vi I I ; possible data from a near-by state . i I [ 140 I I Table 1 gives approximately field-dressed weights of deer I i/ ! I j ! i I : 13o ! I ! based 011 heart girth data from Nebraska, Virginia, Pennsyl­ / I i � 12o �-c--- I [ -� vania, and Wisconsin. Actual weights rather than estimated �- I i 11 0 r-tY.. I l b weights should be used when possible. Figure 5 can be used - I V I i I i 10 0 i < ; for estimating live weight of Nebraska white-tails once : i i i--- t- I i o i// I I field-dressedweight is known. : I : , Once approximate weight and ambient temperature has - < 1 I I o i i : "�¥t I been determined, thigh (Figure 6) and head (Figure 7) : I a _ I �.� V i i Ii temperature measurements can be utilized to determine time I I i I , ! o ! I i I I ; I of death (Table 2). a : � : 40 50 60 10 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 Cooling of a carcass can be affected in many ways (Gill FIELD DRESSED WEIGHT (LB.) and O'Meara, 1965: Moore. 1979). These include dressing, Figure 5. Live weight of white-tailed deer based on field­ transportation and storage. Measurements of both the thigh dressed weight of Nebraska deer. and the naso-pharyngeal temperatures may not compensate

Figure 6. Diagram depicts insertion of two types of thermometers. Sensor should be placed in center of muscle mass in thigh. Thermometer is inserted into the center of the muscle mass from the inner side of thigh . Estimate of center of thigh may be obtained by pushing thermometer clear through thigh, then reinserting the thermometer half that distance. For alcohol thermometers using an angle of 450 to the inner surface of the thigh may facilitate insertion. Meat-type thenno­ meters can be inserted perpendicular to the inner thigh.

18 published in Biometrics and the Wildlife Society Bulletin, respectively. Dr. Woolf worked with eviscerated white-tailed deer that were dead for less than 10 hours. He measured both nasal and thigh temperatures, which enabled him to estimate time of death within 150 minutes with a 95% confidence limit. Kienzler, et a1. (1984), used post-mortem thigh temp­ eratures of white-tailed deer that had been dead for 18 hours or less to derive a mathematical model to determine time of death. His 1983 manual employs thie model to generate tables for dressed deer weighing between 25 and 220 pounds at temperatures ranging from 5 to 650 F. Most research has been conducted on white-tailed deer, but some of this data could have some value when applied to mule deer and black-tailed deer. Reed and Bowden (1974) and Carleson and Kistner (1982) have examined these species.

ELECTRICAL STIMULUS Figure 7. Diagram depicts insertion depth for naso-pharyn­ Using the apparatus shown in Figure 1, several muscle geal thermometer and measurement of pupil diameter. groups were electrically challenged to determine muscle Thermometer is inserted into a nostril and gently pushed response. Locations selected for deer were eye, muzzle, ear, in a direction parallel to the lips. Long-stemed dial­ front leg above elbow, back below vertebrate, tail and type thermometers are recommended for this measure­ exposed inner thigh (Figure 8). Tongue and exposed flank ment. Sometimes obstructions in the nasal passage, such were also possibilities. as bot fly larvae, may cause a problem when inserting Seven positions were routinely checked on Nebraska the thermometer, but they may be overcome by clearing white-tailed deer. Response ranged from very good to none the passage with a stick or some other tool. Make abso­ and are presented in Table 3. lutely certain that the thermometer goes all the way Illinois also did some work with electrical stimulus. In an until it hits the back of the cavity which means the effort to make things easier for officers they reported bulb will be approximately the center of the head. in whether or not electrical stimulus was observed (Table 4). Response to electrical stimulus varied from muscle group for all these variables but may help ascertain how the carcass to muscle group. No "very good" response, except for the was handled. The following observations of Gill and O'Meara (1965) may aid data interpretation. (1) Head normally cooled faster than the thigh. Air Weight (2) Delayed dressing had little effect on head tempera­ No. Temperature OF Pounds ture but slowed thigh cooling. (3) A carcass that was washed out with cold water or 2-1 10-25 60-64 spread-eagled on a surface colder than ambient temperature 2-2 10-25 85-109 may exhibit an increased thigh cooling rate that may even 2-3 10-25 110-134 exceed the head cooling rate. 2-4 20-35 35-59 (4) Head temperatures are rapidly reduced when carried 2-5 20-35 60-84 on top of a vehicle with head forward; the thigh temperature 2-6 20-35 85-109 may not be drastically affected. 2-7 20-35 110-134 (5) Skinning the hams or splitting and propping open 2-8 30-45 35-59 the pelvis should speed thigh cooling but not affect the head 2-9 30-45 60-84 temperature . 2-10 30-45 85-100 (6) Wet deer should cool faster than dry deer. 2-11 30-45 115-140 (7) Deer hung indoors may be affected by the temper­ 2-12 30-45 166·190 ature differences from floor to ceiling. Investigators should 2-13 30-45 195-219 consider all possible variables when consulting Table 2 2-1 4 35-50 110-134 (1-21). The tables are presented in ascending order by both 2-15 35-50 141-165 ambient temperatures and weights. One may choose the 2-16 40-55 35-59 appropriate table to suit their particular situation. If possible, 2-17 40-55 85-109 take a reading once every hour for several hours to enhance 2-18 45-60 140-164 estimate accuracy. If possible, use in conjunction with 2-19 50-65 35-59 electrical stimulus, physical changes in the eye and rigor data. 2-20 50-65 75-99 Additional research on post-mortem temperature changes 2-21 50-65 100-125 in deer has been conducted by Jim Kienzler of the Wildlife Research Station in Boone, Iowa, and Dr. Alan Woolf of the Table 2. Time of death estimates on white-tailed deer based Cooperative Wildlife Research Laboratory , Southern Illinois on weight and post-mortem changes in thigh and nasal University. Results of their research are anticipated to be ternperatures .

19 I\) o

Field-Dressed Weight Field-Dressed Weight Field-Dressed Weigh.

- lb Ibs. 60 84 •. 85 -109 110 - 134 lb•.

HOURS AFTER DEATH HOURS AFTER DEATH HOURS AFTER DEATH " SO 0 10 20 30 40 50 0 10 20 30 40 0 10 20 30 40 50 ' 110 110 1101 1110 • 100 I100 • ''' 100 100 100 1100 � • " . , • •• • t. 901 90 • ..... 90 .,J 90 90 .... 190 • . : ...... : .'fI• • • • 80 .. . 80 80 • 80 80 � 80 1 0 � • ::- • 0 • � � • • '" • I , � • '" '" 2 • •• ....:::> • :::> • � � .. � • • • ...70 , . 70 ...70 • I 70 ...� 70 1 70 �:IE :IE � �:IE • • .... • , .... •• .... • • • • • rI' • , • • • 601 60 60 60 60 1 60 • • • • • • • • • • • • • • • • SO • 501 50 50 50 • • 1 50 • • • • • • • • • • • • • • • •• • • • • • · � • • • • • • • ::I • I:: • ::I I:: • I:: • ::I • II1II . SO 0 10 HOURS20 AFTER DEATH30 40 0 10 H OU20 RS AFTER DEATH30 40 50 0 10 HOURS20 AFTER DEATH30 40 50 Thigh Temperature Thigh Temperature More Than One Observation Thigh Tem perature Thigh Temperature More Than One Observation Thigh Temperature More Than One Observation • o • o • Thigh T em perature o Head Tempera.ore Head Temperature Mor. Than One Observation Head Tempera.ore Head Temperature Mar. Than One Observation • o • 0 • Head Temp.rolare 0 Head Temperature Mar. Than One Observation Field-Dressed Weight Field-Dressed Weight Field-Dressed Weight

lb lb 35 -59 •. 60 -84 lb•. 85 - 109 •.

HOURS AFTER DEATH HOURS AFTER DEATH HOURS AFTER DEATH o o o 10 20 30 40 50 10 20 30 40 50 10 20 30 40 50 1101 1110 101 '110 1101 1 110 • �• �• 100 100 100 100 100 • 100 � I .... • • .... - • ... �� .. • 90 90 V 90 901 90 90 ...: f , t 0 " •. • � • • '.�. • •• ... . • •• 80 . 80 .a � • 80 801 . 80 0 80 � • � • • � . � o � • o . ... � � '" � .. : ,. J. .. :::> :::> => . � 0 • � =t • =t 0 · • • :t : 0 �.. e o � �"- 70 ;'" 70 70 � 70 70 70 • �:IE � � . .. . . � � t· � ... -. � • • . . • • rI' ...... o• • • • 60 • 60 60 60 •• 60 • . 6C • • • • �1' i.� • • • • e ... � so • so 50 50 50 • •• · • 50 • , t • .,. • • .. . . . • - . • • •• . , • • • . • • 40 • •• 40 ' ..: 40 40 1: • • 40 40 • • • • • a • • • • :· • • II . .... • I • • ...... i • . • • • . - 301o 130 301o 130 301o 130 10 20 HOURS AFTER DEATH30 40 50 10 HOUR 20 S AFTER DEATH 30 40 50 10 HOUR 20 S AFTER DEATH30 40 50 Thigh Temperature 0 Thigh Temperature More Than One Observation Thigh Temperature 0 Thigh Temperature More Than One Observation • • Thigh Temperature 0 Thigh Temperature More Than One Observation • • Head Temperat"re 0 Head Temperature More Than One Observation • Head Temperature 0 Head Temperature More Than One Observation • Head Temperat"re 0 Head Yemperature More Than One Observation

N N N

Field-Dressed Weight Field-Dressed Weight Field-Dressed Weight

110 - 134 lb•. lb 35 -59 •. 6() -84 lb•.

HOURS AFTER DEATH HOURS AFTER DEATH HOURS AFTER DEATH o v u 10 20 30 40 50 i 10 20 30 40 50 Ii 10 20 30 40 50 1101 1110 1110 1110 I

100[ 100 100 • 100 .::Jd". - . " 90 � 90 90 90 ... • . •a: ' ( - • . 80 • 80 .. 80 80 o � •• � � . " . . � , � � • � '" •-. '" . :::l '" ;:? :::l � • :t .4b.:J. -.� ::i · ... 70 70 ... 70 · r 70 ::i • ...:( 70 70 •, � � ... � o . -: . . •

• ' •I..; =. • • 60 60 60 • It 60 60 60 1 . • 1 • • • • JJ ...... 1, . - 0 .. • • •• • • • • . 0 • . .. • • . ·· ·11 50 • 5C 50 · • 50 · · 50 50 • :i.· I Ie . 1e '. . C ·: . : . . 0 • I. , � . . . . � . . •� � .-, • . • , • 4' • • 40 40 • • 40 40 • • 40 • • • • •

301o 131 301o 130 o HOURS AFTER DEATH 301 130 10 20 30 40 50 10 20 30 HOURS AFTER DEATH 40 50 10 20 30 HOURS AFTER DEATH 40 50 • Thigh Temperafure 0 Thigh Temperatur. Mor. Thon One Obsetvatiol Thigh Temperature 0 Thigh Temperafure Mot. Thon One Observation • • Thigh Temperature 0 Thigh remperafure Mor. Than One Observation Head Temperator. 0 Head Temperature Mot. Than One Observotio • • Head Temperotor. 0 Head remperature Mor. Than On. Observation • Head Temperator. 0 Head remperature Mot. Than One Observation Field-Dressed Weight Field-Dressed Weight Field-D.essed Weight

lb 85 -109 lb•. 115 -140 •. 166-1 90 lb•.

HOURS AFTER DEATH HOURS AFTER DEATH HOURS AFTER DEATH

10 20 30 40 50 0 10 20 30 40 50 0 10 20 30 40 50 110 110 110 1101 1110 1 1 • • • 100 1100 100 100 • • 0 • • ' o. : .1 : • 90 90 90 I 190 • t • :.-' I :. • � • • 80 • 80 80 80 80 ' 80 9 . 1 ::- • � :'" � • . '" • • � � .-..; . ::> • . • � .... , 70 • 70 � 70 ... 70 �.. 70 70 ::;:( 1 ,;.;. ... • • � � •• • � .. � • . • • � . : 2- - • • • ° ,I· •. • 60 • 60 601 . · 60 60 • • 60 1 -& • �. . • • • • x0 •·• • • • SO • SO 501 • 50 • 50 1 50 • • • • •• .. • • • • • • .. .. • • ' .. • • . • • • • • • 401 • • 40 40 • 40 40 • 1 40 • � • • • • 30 30 30 30 301 130 0 10 HOURS20 AFTER DEATH30 40 50 0 10 HOURS20 AFTER DEATH30 40 50 0 10 HOURS20 AFTER DEATH30 40 50 Thigh Temperature Thigh Temperature More Than One Observation Thigh Temperature More Than One Observation Thigh Tem perature Thigh Temperature More Than One Observation • o • Thigh Temperature o • o • Head Temperatore 0 Head Temperature More Than One Observation e Head Temperatore OHead Temperature More Than One Observation eHead Temperatore OHead Temperature More Than One Observation

N W N """

Fi.ld�Dr.ss.d Weight Field-Dressed Weight Field·Dressed Weight

- lb lb 195 219 •. 110 - 134 •. 141 - 166 lb•.

HOURS AFTER DEATH HOURS AFTER DEATH HOURS AFTER DEATH

.. .. �- o so o SO o SO 10 20 30 40 1 o 10 20 30 40 I 10 20 30 40 110 1 110 10 11 10

• • .. i+ 1 ----. 100 00 100 • 00 100 00 • • I I • • I • ' �: .. -- • ) L , o I 1 + I -+ I----- I . i t 9 -;) I 90 90 • 0 90 I 90 I j ' r 90 I • J' I I • I • •• I • ---1I •• I • I I • • ; • I I • . . -- I I o - I --- I I ... . 1 • • � �.. I" I • t 80 • 80 · 80 80 •• I 80 � • I � · � I o I i o • 0 I � •• � • • , � • I '" '" • , '" I :> • €J :> :> • • • .... - .... I I 70 • - 1 7 :c - + . . - +- - , , � • • • � • , -- -+-- - ; • 0 �... 70 70 70 I 70 ... • • • I ... I I • • � • • • • � � • • I I I • •• • • I - • -+ - 1 --- e · _. +- ) .- 1 -. -- I I - , I I . t �. 60 60 60 i 60 60 • .--+-- I 60 ' I I: .• • ( .. I • I • • .. • I • • I ,-� I r SO SO SO ) L SO� • - -� 50 • .. t- •• • I • .L --�- I -- I 40 40 40 I 4 4 40 • I

) __ . - I 3 o SO 30 30 o .30 3 o 30 10 HOURS20 AFTER DEATH30 40 10 HOURS20 AFTER DEATH30 40 50 10 HOURS20 AFTER DEATH30 40 50 • Thigh Temperatur. 0 Thigh Temperature Mor. Than One Observation • Thigh Temperatur. 0 Thigh Temperatur. Mor. Than One Ob.ervation • Thigh '.mperature 0 Thigh Temperature Mor. Than One Observation 0 • Head 'emperator. Head Temperatur. Mor. Than One Observation • Head Temperator. O Head Temperatur. Mor. Than One Observation • Head 'emperator. 0 Head '.mp.ratur� Mor. Than One Observation Field-Dressed Weight Field-Dressed Weight Field-Dressed Weight lb lb 35 -59 •. 85 - 109 •. 140 - 164 lb•.

HOURS AFTER DEATH HOURS AFTER DEATH HOURS AFTER DEATH

0 10 20 30 40 50 0 10 20 30 40 50 0 10 20 30 40 50 1101 1110 110/ 1110 1101 110 • 1 •

• � .. • 100 100 100 1100 1001 • \ -• oo':l ''' • • • 90 90 • 90 90 • 190 • • •• 'i' • ?i.• • 80 80 80 ...... • 80 80 • 1 80 � � . � . � • � � � . � '" • '" '" • '" . ::> , .. ::> ::> ( . ::> • • • ..... :t • ::i • • • I>.;::i I>.; rI' I>.::i 70 70 I>.::i; 70 70 70 1 70 . �� . . .. :I: �:I: .. ...� ... • � • � !,: I � • ... • • • • • • • • • • • . • 601 . 60 60 , 60 60 60 . , . • 1 . • • 1 ... • .. • • • • • • • • • • , • .. • • • 501 .. • 50 50 • / 50 501 1 50 • • • • • • • • , . • • • � , • • 40 40 40 401 • • 1 401 1 40 • • • 1 • • 1 1 1 30 30 301 130 301 130 0 10 HOURS20 AFTER DEATH30 40 50 0 10 HOURS20 AFTER DEATH30 40 50 0 10 HOURS20 AFTER DEATH30 40 50 Thigh Temperature More Than One Observation Thigh Temperature Thigh Temperature More Than One Observation Thigh Temperature Thigh Temperature More Than One Observation • Thigh Temperature o • o • o Head Temperatore o Head Temperature More Than One Observation . Head Temperatore O Head Temperature More Than One Observation .Head Temperatore OHead Temperature More Than One Observation •

N U1 N 0'

Field-Dressed Weight Field-Dressed Weight Field-Dressed Weight

lb 35 -59 •. 75 -99 lb•. 100- 125 lb•.

HOURS AFTER DEATH HOURS AFTER DEATH HOURS AFTER DEATH

SO SO SO 0 10 20 30 40 10 20 30 40 0 10 20 30 40 1101 1110 110 1101 1110 •• • � " 1001,,_ 100 100 1100 - 100 " . ' " -- . 1':1 90 90 ...- - 90 90 190 .. "r::� • •• 1 '= . , - - I , -- • O · • . 80 • 80 80 • 80 . 80 · � 80 � .� . 1 � • � , , • � 0 � � � 4- • � . � '" • '" � '" • :::> '" '" :::> • :::> 2 . :::> • :t !;( : • II '" � • !;( � � • 70 � • • ... 70 • ... 70 70 � ... 70 1 70 ...... • �:E �:E • • �:E .... • � � -: .... • .... • • • • • 601 1 60 601 - 60 60 1 60 • • • s ol • I SO sol I SO I so 501

40 40 40 40 40 40

o o 301 SO130 301o SO130 301 SO130 10 HOURS 20 AFTER DEATH 30 40 10 HOURS 20 AFTER DEATH30 40 10 HOUR 20 S AFTER DEATH 30 40 Thigh Temperoture Thigh Temperature More Than One Observation Thigh Temperature Thigh Temperature More Thon One Observation • 0 • Thigh Temperature 0 Thigh Temperature More Thon One Observation • 0 - 0 eHead Temperator. OHead Temperature More Thon One Observotion -Head Temp.rator. OHead Temperature Mar. Than One Observation Head Temperatore Head Temperature Mar. Than One Observation A E

Figure 8. Schematics of a deer carcass with electrode positioning marked to denote possible test locations.

27 Time Observed Responses for Back Time Observed ResQonses for Tail Interval Very Interval Very In Hours N Good Good Fair Poor None In Hours N Good Good Fair Poor None

0-0.5 3 3 0-0 .5 3 3 0.51-1.0 15 3 6 4 1 0.51-1.0 13 7 6 1.01-2.0 18 7 2 8 1.01-2.0 29 12 14 2 2.01-3.0 9 2 7 2.01-3.0 16 1 7 7 3.014.0 6 6 3.014.0 7 2 5 4.01-5 .0 3 3 4.01-5.0 3 3

Time Observed Responses for Front Leg Time Observed Responses for Ear Interval Very Interval Very In Hours N Good Good Fair Poor None In Hours N Good Good Fair Poor None

0-0.5 '3 2 1 0-0.5 3 3 0.51-1.0 7 2 2 2 1 0.51-1.0 15 5 2 8 1.01-2.0 20 2 18 1.01-2.0 29 2 13 10 4 2.01-3.0 16 16 2.01-3.0 17 3 4 4 6 3.014.0 6 6 3.014.0 13 1 2 10 4.01-5.0 3 3 4.01-5.0 13 13

Time Observed Responses for Eye Time Observed Responses for Muzzle Interval Very Interval Very In Hours N Good Good Fair Poor None In Hours N Good Good Fair Poor None

0-0.5 3 3 0-0 .5 3 3 0.51-1.0 15 6 6 3 0.51-1.0 15 6 7 2 1.01-2.0 29 2 14 13 1.01-2.0 29 14 12 1 2 2.01-3.0 17 4 3 9 1 2.01-3.0 18 5 5 5 3 3.014.0 8 4 4 3.014.0 9 1 3 5 4.01-5.0 8 2 6 4.01-5.0 7 7

Time Observed Responses For Inner Thigh Interval Very In Hours N Good Good Fair Poor None

0-0.5 3 2 1 0.51-1.0 11 2 4 4 5 1.01-2.0 27 5 6 5 11 2.01-3.0 15 1 3 11 3.014.0 6 1 5

Table 3. Electrical stimulus results are presented for six different time intervals and seven different muscle groups. Response was recorded as being very good, good, fair, poor and none. eye and ear, was observed in times longer than one hour after discrepancies recorded. Use the side with the least amount of death. No response was common for tests conducted 4 hours rigor or the greatest response . after death. After 3 hours, no "good" or "very good" responses were observed Electrical stimulus testing provides an estimate of time of death for deer that have been dead MUSCLE Time In Thigh Flank Hip Eye for less than four hours. Remember, when using the tech­ Minutes N Yes No Yes No Yes No Yes No nique available to us; once "no reaction" has been observed for a specific muscle group, a reaction will not be obtain­ 0-5 9 11 8 3 11 0 11 0 11 0 able from that muscle group at a later time. Electrical 60-90 22 5 17 19 3 19 3 20 2 91-1 19 15 0 15 6 9 12 3 13 2 stimulus can be an excellent supplemental tool when com­ 120-179 12 0 12 4 8 5 7 8 4 bined with the rigor sequence of Gill and O'Meara (1965 180-239 21 0 21 3 19 3 19 11 10 and with pupil diameter and luminosity. ) 240-300 18 0 18 1 17 0 18 0 18 Electrical stimulus tests can be conducted on both sides of the deer and results may vary, depending on how the deer has lain, how it was handled, or where it was shot. When Table 4. Muscle response to electrical stimulus in Illinois possible, both sides of the deer should be checked and white-tailed deer. (Courtesy John Will).

28 HOURS SINCE DEATH D e egre of 13- 1 9- Joint Rigor 4 8 0 2 3 5 6 7 9 10 11 12 18 30

Jaw None Indiana 4 14 3 Maine 0 0 0 Total 4 14 3 Partial Indiana 23 7 Maine 6 6 Total 29 13 Full Indiana 1 14 51 50 31 27 21 29 12 9 2 1 0 2 2 Maine 0 1 18 28 24 14 18 9 8 10 6 1 14 25 13 Total 1 15 69 78 55 41 39 38 20 19 8 2 14 27 15 ------...------_ ..._------_ ...._ ------..------...

Neck None Indiana 5 36 20 1 1 1 1 0 0 0 Maine 0 4 6 6 1 0 0 1 1 1 Total 5 40 26 7 2 1 1 1 1 1 Partial Indiana 10 29 18 8 3 5 6 0 2 0 0 0 0 1 Maine 0 16 16 28 14 20 5 6 4 5 1 4 5 2 Total 10 45 34 36 17 25 11 6 6 5 1 4 5 3 Full Indiana 5 22 22 21 18 22 11 7 2 1 0 2 1 Maine 4 4 10 13 17 11 11 14 7 3 12 27 14 Total 9 26 32 34 35 33 22 21 9 4 12 29 15 ------_ .. ..._ _------...------_ ...... _---_ -.. ------..------Wrist None Indiana 5 41 36 15 5 2 0 1 Maine 0 7 23 15 8 2 2 0 Total 5 48 59 30 13 4 2 1 Partial Indiana 3 18 26 18 16 17 22 8 5 2 1 0 0 1 Maine 0 5 24 37 33 36 11 8 8 1 0 2 4 3 Total 3 23 50 45 49 53 33 16 13 3 1 2 4 4 Full Indiana 2 8 4 3 7 7 1 3 0 0 2 1 0 Maine 0 1 5 4 15 9 16 21 15 11 25 45 25 Total 2 9 9 7 22 16 17 24 15 11 25 47 26

------...------_... _------

Ankle None Indiana 5 39 28 3 3 1 Maine 0 3 3 1 0 0 Total 5 43 31 4 3 1 Partial Indiana 5 22 17 6 3 6 4 1 3 Maine 1 12 14 11 8 3 1 1 0 Total 6 34 31 17 11 9 5 2 3 Full Indiana 12 26 19 18 19 25 10 6 2 1 0 2 2 Maine 2 12 16 16 18 7 6 7 3 3 5 11 4 Total 14 38 35 34 37 32 16 13 5 4 5 13 6 ------_... _------...------_ ..._------.....

Elbow None Indiana 5 33 14 3 1 Maine 0 7 10 0 0 Total 5 40 24 3 1 Partial Indiana 13 23 14 5 4 2 0 0 1 Maine 0 12 20 12 10 5 3 3 4 Total 13 35 34 17 14 7 3 3 5 Full Indiana 2 23 38 24 21 25 29 13 8 2 1 0 2 2 Maine 0 3 20 38 29 49 12 28 22 20 12 25 51 32 Total 2 26 58 62 50 74 41 41 30 22 13 25 53 34

...------...------...--- -

Knee None Indiana 5 34 18 3 Maine 0 4 2 0 Total 5 38 20 3 Partial Indiana 12 21 14 4 2 2 0 1 Maine 1 12 5 3 1 2 1 0 Total 13 33 19 7 3 4 1 1 Full Indiana 2 22 37 24 24 25 29 13 8 2 1 0 2 2 Maine 0 3 20 21 18 20 5 6 7 4 2 5 10 6 Total 2 25 57 45 42 45 34 19 15 6 3 5 12 8

------...... _ _---_ ... ------..---- ..------

Table 5. Rigor mortis observations in white-tailed deer, based on data from Indiana and Maine (Gill and O'Meara 1965).

29 70% 80% 90% 95% 99% Joint None Full None Full None Full None Full None Full

" Jaw 0 2 0 3 3 3 Neck 1 8 1 11 0 0 0 Wrist 2 10 1 10 1 11 0 0 Ankle 1 5 1 6 1 10 0 10 0 10 Elbow 1 4 0 6 0 6 0 10 0 10 Knee 1 3 0 4 0 5 0 7 0 8

rime after death in hours when either no or full rigor was found in of the Maine (Gill and O'Meara, 19 5) Table 6. 70·99% 6 and Indiana deer.

Degree of HOURS SINCE DEATH

19· 0 Joint Rigor 1 2 3 4 5 6 7 8 9 10 11-18 3

Jaw None 23 15 5 Stiff 10 37 18 15 7 14 5 2 9 37 Neck None 30 28 3 2 Stiff 6 26 25 15 4 16 4 4 2 8 40 Wrist None 34 40 12 5 2 1 Stiff 0 1 10 9 5 13 4 2 9 38 Ankle None 31 39 6 2 Stiff 0 17 22 15 7 15 2 2 9 37 Elbow None 34 36 10 4 1 Stiff 0 16 14 14 12 7 4 3 4 10 35 Knee 1'\on e 26 25 3 0 Stiff 0 30 27 17 7 15 5 2 9 3 7

Table 7. Rigor mortis observation in deer from Nebraska (based upon rigor or no rigor present).

70% 80% 90% 95% 99% None Rigor None Rigor None Rigor None Rigor None Rigor

Jaw 3 4 4 4 4 Neck 1 3 3 3 7 7 Wrist 2 5 2 6 2 6 2 6 6 Ankle 1 3 4 1 5 1 5 5 Elbow 1 4 8 1 8 1 8 8 Knee 1 3 3 1 3 1 4 4

Table 8. Time after death in hours when either rigor or no rigor was observed in 70 to 99% of Nebraska deer.

RIGOR MORTIS that may aid in determining TOD. Rigor data includes information from Indiana, Nebraska Rigor was challenged by gently attempting to flex and Maine. Maine and Indiana viewed rigor as none, partial selected joints in their normal plane of movement, accord­ or stiff, while Nebraska viewed it as none or stiff. To com­ ing to Gill and O'Meara (1965). Results are presented in pare data (Tables 5 and 6), time after death was handled Tables 5-9. Joint data agrees somewhat with Gill and similarly for Maine and Indiana (for example, 2 hours should O'Meara's (1965) data for rigor sequence. Normal sequence be interpreted as between 1% to 2% hours). Nebraska data was jaw, then knee, elbow, ankle, neck and wrist, in that was based on whether the joint was stiff or not (Tables 7 and order. 8), and time after death was also handled differently (for When examining deer flex test for rigor on both sides example, 2 hours should be interpreted as greater than 1 of the deer and use the most advanced stage of rigor for hour but no more than 2). In both sets of data there is a estimations. Rigor can be affected by factors which may chance of inconsistency because of the number of people have to be considered when examining deer. collecting data, their interpretations, and the actual time of Rigor may proceed faster than expected when influ. death being supplied by the hunter. T£ble 9 provides an enced by high ambient temperatures, by quick death under insight into several para meters for adult white-tailed deer extreme exertion or stress and by brain damage. Freezing can

30 Temperature Eye Rigor Electrical Stimulas o Ambient in F Pupil Diam. Front Hours Temp. FO Thigh Nasal mm. Color Jaw Neck Wrist Ankle Elbow Knee Muzzle Eye Ear Back Leg Tail Thigh

0 20-35 100 or 97 or may depend fully none none none none none none very very very very very very very greater greater on prevailing transparent good good good good good good good 50-65 100 or 97 or light intensity fluids greater greater at death brilliant fully dialated green to shortly after bluegreen death 13-16 lumenescence almost round

20-35 98 or 80-1 00 13 or greater 75% none none none none none very very very very good good very greater chance of or or or or or good good good good to to good 50-65 100 or 95 or none or partial partial partial partial partial to to to to none fair to greater greater partial rigor rigor rigor rigor rigor fair fair fair none poor

2 20-35 90 or 83-97 11 80% 85% none 80% 65% 70% good good good good good good good greater or chance of chance or none none none to to to to to to to 50-65 95 or 82-96 greater full of no or partial or or or poor poor poor none none fair none greater rigor partial rigor partial partial partial rigor rigor

3 20-35 85-99 76-90 11 full 85% none partial partial partial fair poor poor none none fair poor or rigor partial or or or or to to to to to 50-65 90-100 77-91 greater or full partial full full full none none none none none

4 20-35 83-97 71-85 10 full partial 85% partial partial partial poor ' poor poor none none poor none or rigor or none or or or or to to to to 50-65 84-98 73-87 greater full partial full full full none none none none none

6 20-35 75-89 63-77 9 partial to or full 50-65 75-89 65-79 12 rigor

8 20-35 69-83 51-65 7 luminosity 75% full full full to fades full rigor rigor rigor 50-65 68-82 60-74 10 color rigor grays

10 20-35 65-79 47-6 1 t partial 80% to or full 50-65 62-76 56-70 8 full rigor rigor rigor

12 20-35 60-74 43-52 5 luminosity partial 90% to fades even or full 50-65 62-76 52-66 8 more full rigor color rigor more gray

18 20-35 47-61 37-5 1 luminosity 80% full to ceases full rigor 50-65 55-69 50-60 4 color dull rigor gray

24 20-35 36-50 33-47 no luminosity 80% full to color full rigor 50-65 51-65 50-60 4 still gray rigor

Table 9. Expectations for the majority of yearling adult white-tailed deer that have been dead for a selected number of hours (see Tables 5-8 for more details.) w be mistaken for rigor (Gill and O'Meara, 1965), especially in Pupil Post-Mortem Interval in Hours Diam. the wrist. They suggest checking dew claws, ears and tail for 0.-5 :59 6:00·1 1:59 12:00·18:59 19:00-31:00 evidence of freezing, since they are not appreciably affected by rigor. 15-16 3 13-14 42 2 Other factors may slow the rate of rigor. Rough handling 11-12 111 6 can reduce, delay or even eliminate rigor, depending on the 9-10 87 20 2 state of rigor during the rough handling. Gill and O'Meara 7-8 61 41 3 (1965) found that if an animal was handled roughlywhile 5-6 16 36 5 11 3-4 5 57 18 6 rigor was forming the process could either be delayed or not 1-2 1 21 19 21 completed at all. If rigor was broken once completely estab· 0-1 1 3 lished, it normally did not reappear. If an animal was dragged by the neck, reduced or broken neck rigor can be expected. Total 338 183 48 41 Rough handling could also include hanging a deer by the heck or hind legs. In addition, wounds can weaken tissue Table 11. Distribution of fawn white-tailed deer pupil dia­ and delay or even prevent rigor. Such areas should be avoided meters (mm) by post-mortem intervals in deer from when testing for rigor. Maine (Gill and O'Meara, 1965), Illinois (Woolf et a1. In theory, the rigor sequence should be reversed after a 1983a), Virginia, Indiana and Nebraska. period of time due to muscle breakdown (Morrow 1968). This reverse sequence was variable and not very predictable. However, Gill and O'Meara (1965) did not observe relaxation afterthe first few hours, may require direct illumination with in the lower jaw prior to 48 hours. This was indicated by an artificiallight sour ce. visibility of the front teeth , which are normally concealed by Color plate I shows actual changes that take place in a the lips. The outer end of the jaw could be flexed about deer's eye. Photos were taken with a Nikon FM camera, � inch. equipped with a 200 mm lens. The telephoto lens gave results superior to the macro lens in depicting eye luminescence . PHYSICAL CHANGES IN EYES The color sequence depicts a deer whose closure sequence Following death, eye appearance changes from rigor was slower than normal. More typical closure rates can be mortis of the iris muscle and from loss of transparancy and found in Tables 10 and 11. volume of inter·ocular fluids. Rigor mortis of the iris muscles Gill and O'Meara (1965) made the following useful causes pupil constriction which can be measured with a pair observations on deer eyes: of dividers (Figure 7). Pupil diameters (luminescent area) on (1) First half hour after death eye lens and fluidsfu lly deer from Nebraska, Virginia, Maine, Indiana and Illinois transparent, light reflected from within the eye was a bril· (Tables 10 and 11) were measured according to Gill and liant luminous green (we observed more of a blue-green in O'Meara (1965). Nebraska), and pupil size was dependent upon the prevailing Using the eye with the greatest pupil constriction, the light intensity at time of death. Shortly afterdeath , the pupil vertical diameter was centrally measured at right angles to fully dilated as muscles relaxed. the longest lateral diameter (Figure 7). The lateral pupil (2) 30 minutes to 6 hours - lens and fluids remain diameter in an individual deer remains fairly constant. The transparent, luminosity and color may decrease slightly, the vertical diameter progressively decreases after the pupil fully dilated pupil is almost round (as rigor progresses the has initially dilated. Though not measured in these studies pupil gradually flattens). Fine, diagnostically insignificant the lateral diameter may have some value in estimating winkles may be observed on eye surface. pupil diameter at full dilation. Measurements, especially (3) 6 hours to hours color changes toward gray, luminOSity fades, pupillO width may decrease to about one­ half original diameter. Pupil Post-mortem Interval in Hours (4) 12 hours to 18 hours - color fades to dull gray, Diam. 0.·5:59 6:00-1 1:59 12:00-18:59 19:00·3 1:00 luminosity fades away, pupil may narrow to one-third or less of Original diameter. 17-18 8 (6) After 30 hours - color and pupil diameter remain 15-16 78 1 the same. Hazy blue color appeared over brown colored iris 13-14 150 9 11-12 160 24 1 after about 48 hours. Normally on the third to sixth day 9-10 89 40 2 7 after death, the eyeball partially collapsed (depressions in 7-8 33 38 8 9 eye surface or sagging of eye away from eyelids). This 5-6 10 30 16 14 condition was quite variable, hastened by dry wind and 34 3 19 11 13 delayed by rain or high humidity. 1-2 3 6 6 0-1 2 Effects of freezing - Freezing can stop pupil constric­ tion, change pupil progressively from an initial dull gray

Total 531 164 45 50 after frosting to a milk-glass white when frozen solid. Since pupil constriction is halted upon freezing, an estimate of time of death prior to freezing may be made. Table 10. Distribution of adult white-tailed deer pupil diameters (mm) by post-mortem intervals in deer exam­ CHEMICAL CHANGES IN THE EYES OF DEER ined in Maine (Gill and O'Meara, 1965), Illinois (Woolf Pribor and Bates (1976), Johnson et a1. (1980) found et al. 1983a), Virginia, Indiana and Nebraska. the variables of age, time of year, temperature, sex, weight of

32 Adjusted Coefficient Variance Standard F or Multiple of Error of Number Determinations Equation Estimate Estimate

56 0.76 Y = 39.6 + 7X1 22.8 4.8

56 0.88 Y = 57.4 - 14.4X2 12.5 3.5

56 0.90 Y = 35.1 + 2.2Xl - 12.2X2 10.2 3.2

24 0.94 Y = 37.3 - 6.3 log t + 2.2Xl - 7.5X2

Where Y = estimated post mortem interval

Xl = mEQ/I potassium X2 = log e of mg/dl glucose

t = naris temperature in degrees centigrade

Table 12. Multiple linear regression analysis for nasal temperature and vitreous humor potassium and glucose in moose. (Karns and Kerr, 1980; and Karns and Kerr, 1984a).

1300

1200

1100 • • 1000 • • • 900 .. 0 • • • •

Q.,� • • • Q.. ROO • • � • ce ...J • • • "-lJ 700 > • • • •• "-lJ • .. • O. ...J c. :; • •• • • :::l ()QO 0 0 � • • CiS rJ'J • CD • • • • • -< • _ 0 • c. • .. . - - r- 500 • • •• 0 • • - • Q.,0 .. • • • 0 . O . • • • •• .:J OC. O. _ • • 400 ...... • .. . - .. • • .::> «X:Ce 0 0 • 300 CO .::> 0 • cx::e • o • 200 •

100

0

0 20 40 60 80

POST-'iORTE\1 INTERVAL IN HOURS

Table 13. Scattergram of potassium data in the vitreous humor of white-tailed deer tyes versus the post-mortem interval in hours.

33 deer and head position had very little to do with potassium Gremillion-Smith (1983) sampled 197. Our data agreed with concentrations. However, Schoning and Strafuss (1980) did theirs. Errors may have arisen from the uneven distribution find some discrepancies when dealing with ambient tempera­ of potassium in the vitreous humor (prince, 1977), since tures. We also feel that temperature should show some large volumes were not taken and samples were centrifuged effects. Johnson et al. (1980) showed a logarithmic relation­ rather than filtered. It appears that this may sti11 be a usable ship in mule deer in Utah, with the predicted time of death technique in the future. equalling (log P --5 .51)/0 .0145) where P is in parts per Post-mortem glucose levels in the aqueous humor of million of potassium. Success was obtained through imme­ blacktailed deer were good for up to 8 hours (Pex et al., diate centrifugation of the sample. Karns and Kerr (1980) 1983) and the vitreous humor of white-tailed deer for up to found a linear relationship in both moose (Table 12) and in 24 hours (Karns and Kerr, 1984b). Stress was found to play white-tailed deer (Karns and Kerr 1984b) by using a 0.2 an important factor in glucose concentrations in the vitreous micrometer filter and a 2 - 4 ml sample. Their deer data humor (Karns and Kerr, 1984b) but was not mentioned in had not been carried out beyond 60 hours but standard error the Pex et al. (1983) study. Pex et al. (1983) found that was about 5 hours. glucose levels were relatively independent of temperature and Like Karns and Kerr (1984b), we found a linear rela­ followed Table 14 listed below. The 95% confidence intervals tionship (Table 13). Data discrepancy in the information was are represented by the lower and upper estimated values. presented as a scattergram. We found that the potassium Karns and Kerr's data is yet to be published but is slated for concentration increased for at least 70 hours and then tended 1984. Since it deals with vitreous humor is dependent on to level off (not depicted). Like Woolf and Gremillion-Smith stress and must be filtered, the technique has a few short­ (1983), a great deal of variation was found in samples. We comings, but since it may be used for up to 24 hours it may looked at more than 200 samples, while Woolf and fill a valuable niche in TOD measurements.

Glucose, Y-PRED L-PRED U-PRED Glucose, Y-PRED L-PRED U-PRED rug/ IOO rug (Mean) (Lower) (Upper) rug/ IOO rug (Mean) (Lower) (Upper)

5 5.59500 3.63082 7.55918 33 2.76700 0.88059 4.65341 6 5.49400 3.53514 7.45286 34 2.66600 0.7795 3 4.55247 7 5.39300 3.43928 7.34672 35 2.56500 0.67826 4.45 174 8 5.29200 3.34324 7.24076 36 2.46400 0.57680 4.35120

9 5.19100 3.24702 7.13498 37 2.36300 0,47514 4.25086 10 5.09000 3.15062 7.02938 6.92396 38 2.26200 0.37328 4.15072 11 4.98900 3.05404 39 2.16100 0.21121 4.05079 12 4;88800 2.95727 6.81873 40 2.06000 0. 16896 3.95 104 13 4.78700 2.8603 1 6.71369 41 1.95900 0.06650 3.85 150 14 4.68600 2.76317 6.60883 42 1.85800 -0.03616 3.75216 15 4.58500 2.66583 6.50417 43 1.75700 -0.13901 3.65301 16 4.48400 2.5683 1 6.39969 44 1.65600 -- 0.24206 3.55406

2.47060 6;:29540 - 17 4.38300 45. 1.55500 0 . 34531 3.4553 1 -0.4487.5 18 4.28200 2.37269 6; 19131 46 3.35615 6JJ8741 1.45400 19 4.1810.0 2.27459 47 1.35300 -0.55239 3.25839 5.98310 -0.65622 20 4.08000 2.17630 48 1.25200 3. 16012 21 3.97900 2.07782 5.88018 49 1.15100 -0.76024 3.06224 22 3.87800 1.97913 5.77687 50 1.05000 -0.86446 2.96446 23 3.77700 1.88026 5.67374 51 0.94900 -0.96887 2.85687 24 3.67600 1.78118 5.57082 52 0.84800 --- 1.07347 2.76947 1.68191 5.46809 -1.17825 25 3.51500 53 0.14100 2.61225 26 3.41400 1.58244 0.64600 -1.28323 5.36556 54 2.57523 0 1.4 8278 -1.38839 27 3.3730 5.26322 55 0.54500 2.47839 28 3.27200 0.44400 1. 38291 5.16109 56 -1.49374 2.38174 29 3. 17100 1.28284 5.05916 57 0.34300 -1.59927 2.28527 30 3.07000 1.18258 4.95742 58 0.24200 -1.70498 2.18898 31 2.96900 1.08212 4.85588 59 0. 14100 1.81088 2.09288 32 2.86800 0.98145 4.75455 60 0.04000 1.91650 1.99696

Table 14. Prediction of time interval (hours) when glucose levels are known in blacktail deer. Reprinted from the Journal of Forensic Sciences, Vol . 28, No. 3, July 1983 . James O. Pex et al. Copyright, ASTM, 1916 Race Street, Philadelphia, PA 19103.

34 2 hrs. - 15-16 111111 5 hrs. -12- 13 111111

) hrs. -14- 15 111111 6 hrs. - 1 1-12 111111

7hrs . -IO- 11 111111 II hrs.-6-7 111111

Color Plate No. I depicts typical changes that occur in deer eyes at various times after death . Figures beneath photos give til11e of death and average pupil diameter. (See Explanation of Color Plates.) EXPLANATION OF COLOR PLATES The color plates in this publication are not intended to be used alone to determine time of death. They merely illustrate average occurrences. Individuals can be affected by many variables, especially in the wild . The deer illu strated showed a slower constriction rate than expected for an average deer, . so obviously pupil diameters can vary . Nonetheless, this information can be valu­ able, particularly when used with other data. In addition, the photos may help determine if a hunter is telling the truth ab out when a deer was killed . It is easy visually to check eye luminescence, constriction, and rigor. Suspect deer can be checked for temperature and electrical stimulus, if necessary. One word of caution, temperatures above 600F may have unexpected effects on eye constriction. With geese and pheasants, the photos demonstrated minimum changes expected to take place in the eyes. Again, the photos are not intended as a solo T .O.D. tool. However, they can serve as a guide. As noted in the text, a nu mber of factors can speed up changes. These ilustrations may prove helpful when applying a bit of "conservation officer psychology" in cases where you want to convince a suspected violator that you may know more than you actually do. The plates and the book itself may serve this purpose. WATERFOWL

SAMPLES TEMPERATURE California A total of 113 mallards was used in this study. Most Samples of geese were collected by Hugh M. Worcester birds were killed either through gassing with carbon dioxide, of Tule Lake Refuge in 1941. Thoracic measurements were intravenous injection of sodium pentabarbitol or induced made on undrawn cacklers and snow geese and drawn white­ thoracic pressure. Birds were examined for effects of wetting, fronts at ambient temperatures between 40 and 61OF. body weight, position of measuring device and ambient temperature. Colorado The temperature plots were found to be logarithmic Most of the work on waterfowl and one of the few rather than linear. Thoracic temperatures (measured just in published works on time of death of waterfowl was pub­ front of sternum in thoracic cavity 3-inch penetration) were lished by Morrow and Glover, 1967a. Their observations on considered superior to cloacal temperatures (3-inch insertion 113 mallards are presented in detail in this publication. into vent) and were used in the following tables. Both temperatures were considered statistically usable. Cloacal Indiana temperatures decreased more rapidly than thoracic tempera­ In 1983, Larry Rhinehart and his fellow officers tested tures; 1080 F was characteristically the thoracic temperature one hundred large ducks (predominately mallards and black at death. The effects of body weight, wetting and ambient ducks) for response to electrical stimulus. temperature (controlled temperature rooms) are presented in Figures 9-12. Nebraska Relative humidity was measured with a sling psychro­ More than 100 snow geese were collected at Randall meter at the selected temperatures. For 40 , 50 and 600F Schilling Wildlife Management Area at Plattsmouth, Nebraska. relative humidity was 65-70%, 55-60% and 25-30% respec­ Area managers Mike Moore, John Dinan, and Neal Van tively. Winkle, along with several other Commission biologists Smaller wet birds at low ambient temperatures cooled were very instrumental in the collection and preparation of faster than large, dry birds at high ambient temperatures. samples for analysis. Some geese were also collected at Even with tedious care and the best of conditions, a certain DeSoto National Wildlife Refuge. About 25 Canada geese amount of error must be expected. Several factors may have were collected at Clear Creek Refuge near Lewellen in 1981 pronounced effects on the cooling rates in birds. These and 1982 with assistance from Nick Lyman. Primarily include wind chill, amount of time birds are in water, insula­ crippled birds were selected. Samples were collected either tion of bird fr om the open air, and whether birds are par­ by shooting, running down on foot or with 3-wheeled tially or totally dressed. These variables were not examined vehicles. Specimens that weren't killed by shooting were and an officer will have to take them into account when decapitated prior to use. making a TO D estimate. Decapitation may play an important part in both elec­ trical stimulus data and rigor. Both reactions may be hast­ RIGOR OR MUSCLE STIFFENING ened because of the amount of flopping around which is (Quoted from Morrow and Glover, 1967a) normally associated with this technique. The pattern and sequence of muscular stiffening is an important, established aid in human forensic pathology GEESE (Bendall, 1960 and Forster, 1963). It is most reliable for Published texts on time of death in waterfowl are very the relatively short period when rigor is becoming estab­ limited. It is likely that many states, refuges, university lished. The final breaking of rigor seems more indistinct personnel and fish and wildlife personnel have dabbled not and variable . The onset of rigor in different muscle groups only with temperatures, but with several other means of of mallards was observed by "feel". That is, pressure was determining TOD in these species. carefully applied to a muscle area to test its flexibility. Care Reordering bag limits with the point system now in was taken to disturb the normal course of rigor as little as existence or taking of more than one bag limit are common possible. Muscle groups in the bill, neck, legs, and breast violations today. Published work is essentially limited to were observed. For comparison, the degree of stiffening was Morrow and Glovers, 1967 work on mallard ducks. Other described as none, slight,moderate, or rigid. work is more typically like Worcester, (1941), Morrison and After the first four birds were inspected, it became Hank, (1963) or Kimball, (1972). To give one a better obvious that fu ll rigor is established within 1 or 2 hours understanding of some of the goose data, mallard data from post-mortem. Thereafter, observations were concentrated Morrow and Glover (1967a) - (1967b) will be presented. in this general period. The pattern of stiffening observed The following was extracted either fr om the final among these muscle groups varied because the time inter­ report by Morrow and Glover or from their Special Scientific vals required for the establishment of rigor were variable. Report - Wildlife No. 134, Bureau of Sport Fisheries and Muscles controlling the bill often passed fr om initial Wildlife U.S. Department of Interior. Only information stiffening to a condition of fu ll rigor in 10 minutes or less, currently considered usable by a conservation officer will be but in several instances stiffening proceeded over a much presented. longer period. Stiffening of the bill began as early as 22

37 w a: � l­ e::{ a: w a.. � w I- >­ o o £D � W I­ a: o � I I­ en o a..

6 8 10 12 20 HOURS S I NeE DEATH

Figure 9. Post-mortem aging curves as influenced by ambient temperature. Body temperature data were taken from dry mallards whose average weights were approximately 2.6 1bs.

IIO �----�----�----�----�------�----'-----�----�------�----'

!.LIOO � w a: � -­ I- 9 0 ---- « a: w a.. � W 80t------1------\ I- >­ o � 70�·----·--·- � W I- a: o 60 � I I­ en o 50 a..

40�----�----�----�----�------�----��--�--��------�----� 2 4 8 10 12 14 16 18 20

HOU RS S I NeE DEATH

Figure 10. Post-mortem aging curves as influenced by ambient temperature. Body temperature data were taken from dry mallards whose average weights were approximately 2.2 lbs.

38 110

u... � 100 \ w a::: ::J r- « a::: 90 w 0- � W r- 80 >- 0 0 aJ � 70 w r- a::: 0 � 60 I 2.6 r- I bs· gra oo 1180 ms 0 0- 50

40 2 4 6 8 10 18 20 HOURS SINCE DEAT H

Figure 11. Post-mortem aging curves as influenced by ambient temperature. Body temperature data were taken from mallards placed in water of ambient temperature for three minutes immediately following death and whose average weights

were approximately 2.S lbs.

110

! u... 100 I � I !------L w a::: I I ::J r- « 90 --+ a::: w I 0- � Ij w 80 r- r 0 0 aJ 70 � w r- a::: 0 60 � I r- oo 0 0- 50

40 2 4 20 HOURS SINCE DEATH

Figure 12. Post-mortem aging curves as influenced by ambient temperature. Body temperature data were taken from mallards placed in water of ambient temperature for three minutes immediately following death and whose average weights

were approximately 2.2 lbs.

39 minutes and as late as 45 minutes post-mortem. Full rigor becomes blackish-brown to ultramarine-violet at the same was reached as early as 27 minutes and as late as 75 minutes time and has a clear, watery look due to the fluid between 7 post-mortem. In one exceptional case, stiffening of the bill and 12 hours after death. After an indefinite period the was observed to begin after 60 minutes post-mortem and cornea over the iris sinks to the level of the pupil and the terminate in full rigor only after 170 minutes post-mortem. entire eye appears flattened. This seems most characteristic In the muscles of the neck, stiffening began as early as 8 of the period 14 to 24 hours after death. The iris loses its minutes and as late as 45 minutes post-mortem. Full rigor watery appearance at this time and becomes a dull or opaque was achieved as early as 10 minutes and as late as 75 minutes ultramarine-violet; it may appear medium gray to blackish­ post-mortem. In one bird, neck stiffening began at 70 min­ brown. However, the trend is usually toward darker colors as utes post-mortem and ended in full rigor after 140 minutes time since death increases. From the flattened condition of post-mortem. the eye, the pupil becomes increasingly sunken as moisture Muscles of the legs began to show stiffening as early as 8 is lost. The pupil remains generally flat causing the iris to be minutes and as late as 40 minutes post-mortem. Full rigor pulled into an increasingly concave shape as it slopes upward was achieved as early as 10 minutes and as late as 70 minutes to the white sclerotic ring outside the iris. This commonly post-mortem. A period of 20 to 30 minutes was usually begins near 22 hours post-mortem and gradually becomes required in the stiffeningprocess . Stiffening began in the flightmuscles as early as 25 min­ utes and as late as 60 minutes post-mortem. Full rigor was Hours reached as early as 40 minutes and as late as 90 minutes After post-mortem. Death A B C D E F G H I Although time intervals required for stiffening and the sequences observed among parts are variable and conflicting, enough pattern existed to suggest that stress and handling from capture until killing might influence the results ob­ served. Muscular activity was probably an important factor. !\]\i� f� � l i PHYSICAL APPEARANCE OF THE EYES 5 3, 13"'12 10 7 4 (Quoted from Morrow and Glover 1967a) 6 1'13 11 11 7 5 The eyes of dead mallards were found to change in 7 1 13" 11"'1 1 7 7 appearance in a definite pattern. The characters most useful 8 1 11 "9 13 8 7 were color and shape. Poor illumination prevents accurate 9 1 9 8""- 14� 6 9 2 1 observation of eye color so a flashlightwas used to illuminate 10 1 9 7 14, 6 9 3 1 the eyes of all but the first four birds. The eyes were 11 1 8 7 14 "5"'10 3 4 described as to general shape and turgor, shape of pupil and 12 1 5 7 14 4'"16 3 4 iris areas, color of pupil, and color of iris. The eyes of the 13 4 6 16 5 18\3 5 birds were first observed and described one hour after death 14 4 6 16 7 18 5 6 and at hourly intervals. 15 2 6 16 Although an overall pattern was evident, variation 16 2 4 20 occurred. Corresponding changes happened at different 17 2. 1 21 8 20 5 5 post-mortem intervals, and the eye conditions observed 18 2 22 8 20 5 7 were inconsistent in duration. Even so , useful time of death 19 2 22 8 20 6 7 estimates can be made on the basis of eye appearance. 20 2 22 5 20 6 7 The following description of post-mortem eye changes 21 1 22 5 20 6 9 is a composite presentation of all observations of eyes found 22 1 22 4 20 5 11 to be of value in estimating time of death and the intervals 23 1 22 4 20 4 12 at which they are most likely to occur. The range of occur­ 24 1 22 4 20 4 12 rence and characteristics of eye changes are presented in 27 22 22 2 9 Table 15. Color terms follow the Villalobos system (Palmer, 30 22 22 2 9 1962). 33 22 22 1 10 At death and for commonly 2 hours thereafter mallard 36 22 22 1 10 eyes maintain their normal appearance as in the living bird. 39 22 22 10 The pupil is ultramarine and slightly cloudy, the iris is 42 22 22 10 chestnut and the whole eye has a smooth, moist, and turgid 45 22 22 13 appearance. As moisture evaporates from the eye turgor is 48 22 22 13 lost, small wrinkles appear, and indentations begin to form in 54 22 22 1 3 the eye. These conditions typically prevail through about 9 60 22 22 10 3 hours post-mortem. A change occurs in pupil color near 5 to 66 22 22 7 5 8 hours post-mortem ; the pupil becomes turquoise-cobalt, 72 22 22 2 5 with a reflective or glossy appearance from inside the eye ; 78 22 22 2 5 no further changes in pupil color occur. With further moisture loss the corneal surface over the pupil generally Table 15. Observation frequencya/ of certain post-mortem becomes sunken and flat. The cornea remains elevated over characteristics of mallard eyesb/; arrow indicates the the iris as a circular ridge around the sunken pupil. The iris general progression of change.

40 Table 15., Cont. more pronounced. After about 50 hours post-mortem the pupil area around the edges begins to assume the concave a/ Each fr equency unit indicates that the characteris­ shape of the rest of the eye. The flattened area in the center tic(s) was descriptive of one mallard eye at the corres­ of the eye gradually shrinks until the iris and pupil areas are ponding post-mortem interval. Observations for 11 birds uniformly and deeply concave. One or both eyes of a bird are included. Only four were observed fo r 78 hours; two normally have reached this condition after about 60 hours for 58 hours and five for 24 hours. Observation frequencies post-mortem. for characteristics D and F are extended past the 24 and 58 Many eye conditions were not as distinct as those hour intervals (even though data are lacking) because these described, because they were complicated by irregular characteristics seem to persist unchanged for the full period. wrinkling. Since most of the changes described are the result of moisture loss from the eye, evaporation rate has an b / Post-mortem characteristics of mallard eyes: important effect on the time intervals involved.

A. Eyes as in life ; moist, only slight loss in turgor; TEMPERATURE iris, chestnut. Data on TOD in geese appears to be even more limited B. Turgor of eyes much reduced; indentations and than duck information. In 1941, Worcester worked with wrinkling present, but not fully collapsed. white-fronted geese cacklers (probably West Coast Canadas) C. Pupil ultramarine and slightly cloudy. and snow geese at Tule Lake in California. Temperatures D. Pupil turquoise-cobalt with a reflective or glassy were again taken thoracically, instead of cloacally. Results appearance from inside the eye. were presented for undrawn cacklers (8) Figure 13 and snow geese (3) Figure 14 ambient at temperature ranges 560 F E. Corneal surface over pupil sunken and largely flat ; cornea elevated above iris as a circular ridge. and 610 F. His temperatures were not as controlled as F. Iris blackish brown to ultramarine vio let; watery Morrow and Glover's (1967a) and his cooling rate for these between cornea and iris in association with char­ birds was a little slower than theirs. acteristic E, becoming an opaque violet-magenta Both cacklers and snow geese are normally larger than with characteristics G and H. mallards, so this should be expected. Snow geese and giant G. Cornea sunken flat over iris; whole eye is flat. Canada geese were collected in Nebraska. ,Many of these H. Iris is pulled into an increasingly steep, concave birds, especially the snow geese, were either sick or cripples. position as the flatpupil area becomes more Temperatures on these birds would be biased and are not sunken. included in this study. Temperatures (3-inch cloacal inser­ I. Area of iris and pupil a uniformly concave tion) were taken on Canada geese ranging in weight from 6.7 depression. pounds to 12 pounds and between 30 and 400F, Figure 15.

110

100

90

� 0 � 80 ...= ;..� Q) c.. 5 70 f-i

60

.-

50

40 4 10 12 14 16 18 20 Post Mortem Interval In Hours

Figure 13. Worcester's data showing thoracic change in temperature with time in cackling geese.

41 90

60

50

40 4 6 8 10 12 14 16 18 20 Post Mortem Interval In Hours

Figure 14. Worcester's data showing thoracic change in temperature with time in snow geese.

110

100

90

o lJ;.. 80 o � �� 0 := • - o f: 70 Q) !:l. . . S � . Q) � 60 � �� . �� . . 50 o �O o • o

40

30 ·______-W�______4 6 8 10 1� 14 16 18 �O 24 Post Mortem Interval In Hours

Figure 15. Cloacal temperature change in Canada geese, Symbols represent different birds.

42 As expected cooling rates were slower than mallards even CHEMICAL CHANGES IN THE EYE though controlled environment chambers were not available. Chemical changes in a goose's eye are limited presently All Nebraska temperatures were taken cloacally and not to potassium levels in the vitreous humor. Levels were run thoracically like other studies and all birds were dry. jointly on both snow geese (blue geese) and Canada geese, since the current procedure was not adequate to differentiate ELECTRICAL STIMULUS the two species. Results are presented in Figure 17. All Nebraska birds were decapitated, which affected Although not determinable in the field, our data appear both rigor and electrical stimulus testing. Both rigor and to follow a linear rather than a logarythmic path (Morrow electrical stimulus are closely tied together, especially in and Glover 1967a). The F value is 190.6 and the formula for birds. When rigor has set in completely, electrical stimulus the best statistical fit is y = 7.34x + 366.0. Where y is the will probably not be observed. Although rigor was not a potassium level in ppm, x is the post-mortem interval and parameter studied in geese, observations would defintely 366.0 is the intercept of the best theoretical line (least point to the variability of rigor and support Morrow and squares equation). Glover's (l967a) findings in mallards. Indiana decapitated Fewer techniques are available for determining time of one hundred large ducks in 1983. Response to electrical death in geese than deer. Enough information is available stimulus was recorded as being either negative or positive though that one should be able to determine if more than a in the neck, wing, back and leg. Results are very similar to legal bag limit was taken in a single day. Determinations on that observed in Nebraska. Their data is presented below. bag reordering should also be possible in some cases.

Time after Death Neck Wing Back Leg D in Minutes + + + + B 15-18 37 2 39 1 39 1 25 15 20 8 2 10 0 10 0 5 5 25 17 3 1 7 3 17 3 9 11 30 3 7 7 3 6 4 0 10 41-46 3 17 4 16 1 19 0 20

No reaction was observed in these birds after 46 min­ utes. The neck and the wing will most likely respond longer than other muscle groups. We found the head, tail, and wing to respond the longest in Nebraska. parameter studied in geese, observations would definitely point to the variability of rigor and support Morrow and Glover's (1967a) findings in mallards. A few ducks were tested by electrical stimulus but those that were showed no reaction after an hour and usually none after about half that period. For geese the reaction was longer and lasted for up to 1 � hours. Geese were checked at selected body locations Figure 16. Results are listed in order of their failure to react to electrical stimulus. Normal response was breast or back first, then leg and then head, tail or wing, Tables 16 and 17. The wing was usually the last to quit reacting or at least was the easiest to see, primarily due to its length and the position tested. Measurements were usually taken in wind free areas so this interference, which may be common in the field, was normally not present at these measurements.

PHYSICAL CHANGES IN THE EYE Physical changes in the eye should parallel that of ducks. Primary changes are moisture loss and color. Color plate II shows the visual changes at various periods after death. Photos depict a snow goose at 320F for a period of 68 hours. Temperatures lower than this may cause freezing, while higher temperatures and wind would speed up loss of moisture. The pictures should reflect minimal changes in a goose's eye. Morrow and Glover, 1967a describe color changes in a mallard duck similar to what can probably be Figure 16. Locations on a goose where electrical stimulus was expected in geese. checked.

43 Reaction Reaction Time in Very Time in Very Location Minutes Number Good Good Fair Poor None Location Minutes Number Good Good Fair Poor None

Wing 0-15 26 23 1 2 Wing 0-15 4 4 Wing 16-30 38 5 17 9 5 2 Wing 16-30 10 8 2 Wing 31-45 44 9 14 14 7 Wing 31-45 4 1 3 Wing 46-60 47 2 5 22 18 Wing 46-60 9 2 6 1 Wing 61-75 22 1 2 19 Wing 61-75 12 1 5 3 4 Wing 76-90 2 2 Wing 76-90 11 3 8

Tail 0-15 26 19 4 1 2 Tail 0-15 2 2 Tail 16-30 39 3 15 6 1 14 16-30 7 5 Tail 2 Tail 31-45 44 6 7 6 25 Tail 31-45 4 2 1 1 Tail 46-60 46 1 5 2 38 Tail 46-60 8 2 3 2 Tail 61-75 29 2 27 Tail 61-75 10 1 2 6 Tail 76-90 2 1 Tail 76-90 9 9

Head* 0-15 17 16 1 Head* 0-15 4 4 Head 16-30 15 7 3 2 3 Head 16-30 9 8 1 Head 31-45 17 1 2 4 3 7 Head 31-45 5 1 2 1 1 Head 46-60 26 5 6 15 Head 46-60 8 1 2 4 1 Head 61-75 10 1 1 8 Head 61-75 12 1 2 9 Head 76-90 8 1 7 Head 76-90 11 11

Breast 0-15 26 22 4 Breast 0-15 4 4 Breast 16-30 40 3 3 9 1 24 Breast 16-30 10 5 5 Breast 31-45 43 1 5 37 Breast 31-45 3 1 2 Breast 46-60 47 47 Breast 46-60 9 1 7 Breast 61-75 3 3 Breast 61-75 10 1 9 Breast 76-90 11 11 Leg 0-15 24 19 3 1 1 Leg 16-30 39 3 4 8 7 17 Leg 0-15 4 4 Leg 31-45 43 3 7 33 Leg 16-30 11 6 3 2 Leg 46-60 47 2 45 Leg 31-45 4 1 1 1 1 Leg 61-75 30 30 Leg 46-60 12 1 1 3 7 Leg 61-75 11 11 Back 0-15 16 14 2 Leg 76-90 11 11 Back 16-3 0 28 5 5 7 10 1 Back 31-45 33 1 4 28 Back 0-15 4 4 Back 46-60 34 1 33 Back 16-30 10 5 4 1 Back 61-75 22 22 Back 31-45 4 2 1 Back 46-60 11 1 8 *(bill eye) Back 61-75 11 2 9 & Back 76-90 11 11

* (bill eye) Table 16. Snow goose electrical stimulus data showing & reactions of six muscle groups with respect to time.

Table 17. Canada goose electrical stimulus data showing reactions of six muscle groups with respect to time.

44 1500

• 1400

1300

. 1200 • • •

1100

• .0 1000 • <» • • • e 900 Q. • • Q. • • • . 5 • 800 o. �Q,) > • .sa 0 • e 700 • . e • � • c:J / .... • 0 a.. 600 • • 500 • • 400 • • • <» • 300 00. • .0 • 200 • • 100

0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 Post mortem interval in hours

Figure 17. Potassium levels in the vitreous humor goose eyes with respect to time.

45 Color Plate No. 2 illustrates typical changes in goose eyes at various times after death at an ambient temperature of approximately 32°F. Wanner temperatures. lower humidity. and wind should speed up such changes.

2 hrs. 7 hrs.

17 hrs.

14lfl hrs. 32 Y2 hrs.

43 '12 hrs. 68 hrs. PHEASANT (Phasianus colchicus)

TOD data on pheasants is usually limited to roosters and for these birds. Thigh temperatures at 50° F showed a is normally not for long time intervals. It still may be possi­ different relationship, for birds that are both dressed differ­ ble to determine whether a bird was taken before or after ently, and are of different weights (Figure 19). Depending legal hunting hours, or if a person had exceeded daily bag upon handling conditions, the head or cloaca may not be limits. Presently body temperature (1040 F alive) may be present in some field birds. Using 3" insertion, Nebraska did one of the most reliable indicators of time since death. measure cloacal temperatures of 7 birds at an ambient Electrical stimulus, physical condition of the eye and potas­ temperature of 400F, Figure 20. Results can be compared to sium levels in the vitreous humor may also be important. Pennsylvania's data, (Figure 21). Their thigh or breast temperatures were considered very reliable. Figures 21 and PHEASANT SAMPLE COLLECTION 22 show that at the ambient temperatures of 400 and 500 F Nebraska respectively, the thigh cooled the fastest. It also showed the Pheasants were supplied through the courtesy of Don greatest temperature decrease and it was very useful in Studnicka of the Sacramento Wildlife Management Area near determing TOD for up to 4 hours. Its low biomass also Wilcox and cooperating biologists and officers from the cooled quicker and provided the least amount of variability. Nebraska Game and Parks Commission. The rate of temperature decline of the thigh at 3 ambi­ ent temperatures is depicted in Figure 23. As expected, Iowa decline was greatest at the lowest temperature. At the 95% Pheasants were supplied through the courtesy of a State confidence interval, the rate of decrease was greater at 400 F Game Farm and Bob Barratt, superintendent of the Wildlife than at the 600 F level. This was not true of either extreme Division. when compared to the 500 F level. It should be noted that thigh temperatures dropped 150 to 200 F in the first hour Pennsylvania and that most of the temperature loss was in the first 2 Data was provided by the Pennsylvania Game Commis­ hours. sion. Special thanks to biologist William Shope, Hershey Typical cooling curves were observed in two pheasants Medical Pathologist Dr. John Kreider, and the Pennsyl­ with dissimilar weight. Even though the difference appeared vania Game Commission drafting department for graphs real and there was about 3/4 of a pound difference in weight, of the data. no statistical difference in these birds was found at the 95% confidencelevel . TEMPERATURE For short periods of time after death at ambient temp­ The effect of temperature may best be ascertained eratures of 400 to 600 F, Tables 18-20 can be utilized. At by looking at Pennsylvania data on 24 roosters. Birds were 95% confidence intervals one can obtain TOD to within gassed with carbon dioxide and maintained in controlled approximately one hour. For longer periods of time breast temperature rooms of 400, 500 and 600 F respectively. measurements can be taken. At the 95% confidence levels Temperature measurements were then taken every half hour approximately two hours rather than one was observed. In for approximately 24 hours. Some of the birds were gutted order to check results, approximately 60 pheasant thighs and some were left intact. Some birds were wrapped in were measured at Graterford Prison in Pennsylvania. TOD burlap and placed in a hunting coat to simulate field condi­ was supplied by the hunter and most pheasants were carried tions. The other birds were laid on wooden tables in con­ in a hunters vest. Results indicate (Figure 24) that thigh trolled temperature rooms. A metal dial thermometer was temperatures were the best for the first couple of hours. used to measure the temperature in the mouth (approxi­ The temperature data may allow one to estimate a range mately 2-inch insertion) cloaca (2-inch insertion) and in the of time after death rather than an actual time in minutes muscles of the breast (2 centimeter insertion in the center of and hours. muscle mass on one side of breast) and thigh (2 centimeter insertion in the approximate center of muscle mass). ELECTRICAL STIMULUS Some differences in the rates of temperature decline Some electrical stimulus data has been taken both by were noted for eviserated birds vs. non-eviserated birds, for Iowa and Nebraska. Nebraska birds were decapitated, which out of the hunting coat vs. in the hunting coat, and for may account for the discrepancy in the small amount of various bird weights (Figure 18). However, these differences existing data. Iowa found a maximum reaction time of were not great enough to be significant when breast tempera­ about 2 hours. Nebraska found the maximum reaction time tures were taken. An almost straight line decline was noted to be in the wing and to last no longer than about 1 � hours.

49 110

UNGUTTED, NOT IN HUNTING COAT. AV E. WT. 1387 9 (3 1/10 Ib )

UNGUTTED, IN HUNTING COAT . AVE. WT. 12609 {2 4/5 Ib)

GUTTED , NOT IN HUNTI NG COAT. AVE. WT. 1310 9 (2 9/10 Ib)

UNGUTTED , NOT IN HUNTING COAT. AVE. WT. 1033 9 (2 1/3 lb )

�80 " o " I W It: => ....

.... <16(I) 0 w It: CD

50

I HR 2HR 3HR 4HR 5HR 6 HR 7HR 8HR 9 HR ff 22 HR TI HOURS AFTER DEATH ME -

Figure 18. Average breast temperature of male pheasants at 40°F ambient temperature in various situations.

110

II 30 9 (Z liZ I b), UNGUTTED

IZ40 9 (Z l/4lb), UNGUTTED 1270 9 (Z 4/5lb), UNGUTTED

1330 9 (Z !VlOlb ), GUTTED

90 o 1370 9 (3 Ib ), GUTTED

1220 9 (Z 3/4 Ib ) , GUTTED , THIGH UNFEATHERED

80

o

. --:- -:--. -:- .­ .�•

40 80 120 160 200 240 280 320 I HR 2 HR 3 HR 4 HR 5 HR

TIME -- MI NUTES AFTER DEATH

Figure 19. Thigh temperatures of individual male pheasants at 50°F ambient temperature.

50 110

• o 100 V . o !)'.

'.)J.V ... 90

�.V ... . v 'll ... . 80 o V o • V ... . o • "'" A 0 V • 0 • 0 70 V . ... .5 o . ... . � . ::: o • 0 V 0 . ... 0 • V 0 • c.. 0.- • � 60 V 0 • 0 VO o • E..... V 0 o • "<::l 0 0 V o. = • 0 50 V • � 0 V 0 • 0 V' 0 0 • 0 • V V 0 • 0 0 0 • ... V V V • • 40 0 • • 0 0 • • ... 0 • •

3()

10

9 10 11 12 13 14 15 16 11 18 19 20 21 22 23 24 2S 26 27 28 29

Post·mortem interval in hours

Figure 20. Body temperature of adult pheasants taken cloacally at an ambient temperature of 40°F.

110 MALE PHEASANT

La.. o 40 of AM BIENT TEMPE RATURE 90 LIJ a::: :J I­ ct'80 a::: LIJ 0.. 2 LIJ 70 I-

>­ o o 60 CD

50�------�------�----�------�------�------�------�------�------p------� 250 500 41HR HR TIM E - MINUTES DEATH (AVERAGE TI ME )

Figure 21. Average cooling rate at selected locations on male pheasants at 400F.

51 MALE PHEASANT 110

lI. 5 0 0 F • AMBIENT TEMPERATURE I IaJ 10 0:: :l

� 9 IaJ0:: Q. ::E ...I.LI 80

>- g 70 CD

6�------.-----�------��----�------�----�------�------�----�------� 250 300 500 550 I I HR 2 HR 3HR 4 HR 5 1 tfl 6 HR 7 HR 8 HR 91 HR

TIME - MINUTES AFTER DEATH (AVERAGE TIME )

Figure 22. Average cooling rate at selected locations on male pheasants at SOoF.

110 MALE PHEASANT

100

90

. . . . . 0 0 0 0 . . . 80 . . . 0 AT :: AMBIENT TEMPERATURE 0 o. ". ' . . . ' " . . - . . . II.. . -.... 0 . AT o 10 ...... 60 F - . . " . " I . ' . 1&1 . . . C . . . :;, . . � ' . . c 60 ...... ; . . IaJ ·0 . .... ' . . . . . ::c ' . . � . . - . .... 0 • • 40 i 50 0 F-AT

50 I 100 I 150 200 250 I HR 2 HR :3 HR 4 HRI

TIME MINUTES AFTER DEATH ( AVERAGE TI ME )

Figure 23. The rate of decline in thigh temperatures at three ambient temperatures.

52 THIGH BREAST Temp. of Minimum Average Maximum Minimum Average Maximum

109 1 3 108 3 21 107 6 30 106 9 39 105 1 3 47 104 16 56 103 19 6 64 102 23 15 73 101 26 24 82 100 30 33 92 99 33 43 101 98 37 52 110 97 2 40 3 62 120 96 6 44 13 7 1 129 95 10 47 22 81 139 94 1 3 51 32 90 148 93 17 55 42 100 158 92 21 59 52 110 168 91 25 63 62 120 178 90 29 66 72 131 188 89 33 70 83 1 41 199 88 37 74 93 151 209 87 2 41 78 104 162 220 86 6 45 82 114 172 230 85 11 49 87 125 183 241 84 15 53 91 136 194 252 83 19 57 95 147 205 263 82 23 61 99 158 216 274 8 1 28 66 104 170 228 286 80 32 70 108 181 239 297 79 36 74 112 193 251 309 117 78 41 79 204 262 32 1 77 46 83 121 216 274 333 76 SO 88 126 228 286 345 75 55 93 131 240 298 357 74 60 97 136 253 311 369 73 64 102 ]40 265 323 382 72 69 107 145 278 336 395 71 74 112 150 291 349 408 70 79 117 155 304 362 421 69 84 122 161 317 375 435 68 89 127 166 331 389 448 67 94 132 171 344 402 462 66 10Q 138 177 358 416 476 65 105 143 182 371 430 490 64 110 148 188 386 444 505 63 116 154 193 400 459 519 62 121 159 199 415 474 534 6 1 127 165 205 429 489 550 60 133 171 211 444 504 565 59 138 177 217 460 5 1 9 581 58 144 183 223 475 535 597 57 150 189 229 49 1 551 613 56 156 195 236 507 567 630 55 163 20 1 242 524 584 647 54 169 208 249 540 601 664 53 175 214 256 557 618 681 52 182 221 263 574 636 699 5 1 189 228 270 592 654 717

Table 18. Cooling rates (in minutes) of male pheasant thighs and breasts based upon 95% confidence limits and an ambient temperature of 40°F.

53 T H IG H BREAST Temp. of Minimum Average Maximum Minimum Average Maximum

109 13 35 108 16 46 107 19 57 106 22 68 105 26 79 104 29 90 "0 103 J_ 101 1 02 35 113 101 39 1 2 125 '" 100 J 42 24 136 99 6 46 36 148 98 10 50 48 160 97 14 53 60 172 96 1 9 57 72 184 95 23 61 84 197 94 27 65 97 209 93 31 69 110 222 92 35 73 1 0 1 23 235 9 1 40 77 T; 135 248 90 44 82 36 149 261 89 1 48 86 49 162 274 88 6 53 91 62 175 287 87 12 57 95 76 189 301 86 18 62 100 90 202 315 85 23 66 105 104 216 329 84 29 7 1 110 118 230 343 83 34 76 116 132 244 357 82 40 80 121 147 259 372 8 1 45 85 127 161 273 386 80 50 90 133 176 288 401 79 56 95 139 191 303 416 78 61 100 145 206 318 431 77 67 105 151 221 333 447 16 72 11 0 158 252 349 478 75 77 115 165 268 365 495 74 82 1 21 i72 276 380 503 73 87 126 179 284 397 511 72 93 131 186 301 41 3 527

7 1 98 1"'J/'''' 1 93 317 430 544 70 103 142 20 1 334 446 561 69 108 148 209 351 463 579 68 113 154 217 368 481 596 67 118 160 225 385 498 614 66 124 1 65 234 403 516 632 65 129 171 242 421 534 65 1 64 134 178 251 439 553 670 63 139 184 260 458 571 689 62 145 190 269 476 590 708 61 1 50 196 279 495 609 728

Table 19. Cooling rates (in minutes) of male pheasant thighs anJ breasts based upon 9S(k confidence limits and an ambient temperature of 50°F.

54 THIGH BREAST Temp. of Minimum Average Maximum Minimum Average Maximum

109 44 108 55 107 2 66 106 6 77 105 11 88 104 16 100 103 21 111 102 26 122 101 3 1 11 134 100 36 22 146 99 42 34 158 98 47 46 170 97 52 58 182 96 6 58 71 194 95 11 63 83 207 94 17 69 96 219 93 22 75 108 232 92 28 81 121 245 91 34 87 10 134 258 90 40 93 23 147 271 89 46 99 36 160 284 88 52 105 50 174 187 312981 87 4 58 112 63 86 10 64 118 77 201 325 85 17 70 125 91 215 339 84 23 76 132 105 229 353 83 30 83 139 120 243 368 82 36 89 136 134 258 382 81 43 96 153 149 272 397 80 50 102 160 163 287 412 79 56 109 168 178 302 427 78 63 116 175 194 317 443 77 70 122 183 209 332 458 76 76 129 191 224 348 474 75 83 136 199 240 363 490 74 90 143 207 256 379 507 73 97 151 215 272 395 523 72 104 158 224 288 412 540 71 111 165 233 304 428 557

Table 20. Cooling rates (in minutes) of male pheasant thighs and breasts based upon 95% confidence limits and an ambient temperature of 60°F.

PHYSICAL CONDITION OF THE EYE For periods of time longer than a few hours, heads Pictures were taken of pheasant eyes at temperatures were removed and stored in a refrigerator (ambient temper­ ranging from 320 F to approximately 700 F (color plates ature 34-360 F). Like geese , eyes from the same head were III-VI). The warmer the temperature, the faster the decline assumed to have the same potassium levels. Usually two to in eye condition. Condition is dependent not only on temp­ three individual eyes were selected for each time period. A erature but wind conditions, for wind will hasten drying. One graph of both Iowa and Nebraska birds is presented to show should still be able to tell the difference in birds taken a day the amount of variation observed between the two states. apart and determine daily bag limits. A great diffe rence can Eyes were observed for up to 150 hours with no tailing off exist between individual birds. Our birds were essentially free like that observed in deer. The equations based upon pre­ from wind currents but the difference still existed. Color dicted values in Figures 25 and 26 are presented below. For changes and wrinkling of the eye should be similar to that of Iowa, the F value equaled 297, and Y equaled 9.7X + 449 . ducks (Morrow and Glover, 1967a). For Nebraska the F value equaled 245 , and Y equaled 8 .7X +

404 (Y = the potassium level in ppm and X = the post­ CHEMICAL CHANGES IN THE EYES mortem interval). The estimate is always at least 4 hours Experimental evidence with chickens suggested that the greater for the Nebraska data. If this reaction is temperature­ vitreous humor should be taken for geese . Because of the dependent, these results are representative of the least retina, vitreous humor cannot be taken though the sclerotic amount of change in potassium levels. membrane (white of eye).

55 110 VI 0"'1

EACH DOT REPRESENTS THE TH IGH TEMPE RATURE OF

100 • EACH PHEASANT. THE LI NE I S THE AV ERAGE TH IGH ° TEMPERATURE (40 F-AT) FOR THE EXPERIMENTAL PHEASANTS .

•

90 • •

•

80 •

. . • •

� • •

70 • • l&.. '" 0 ...,."",. . . • • • • � . It: • :::;) •• l- 60 • c( • It: • � CL. ::E � ...

50

80 120 200 240 280 320 360 400 40 160 1 2 HR 3HR 41fl 6 HR 5 HR HR TIME MI NUTES AFTER DEATH

Figure 24. Male pheasant data collected at Graterford prison hunt, November 1973. Ambient temperature was32 ° - 42°F. 02 SO I

2000 •

1750

1500

(:l..E (:l.. .5 '" • 1250 • Q)> � E ;:l • 0 'lil '" 1000 • � '0 �

750

500

250

0 20 40 60 80 100 120 140 160 Post-mortem interval in hours

Figure 25. Potassium levels in vitreous humor of Iowa pheasant eyes.

1700

1600

1500 • •

• 1400

• 1300 • • • • • • • 1200 • E • • • (:l.. • • (:l.. 1100 • • 5 • '" • • • Q) 1000 0 > • • • � • • • • • 900 • E • • 0 • • .�� 800 •• - • '0 • • 0 � - 0 0 0 •• • •

•

400 •

300

200

10 20 30 40 50 60 70 80 90 100 110 12l' 130 140 Post mortem interval in hours

Figure 26. Potassium levels in vitreous humor of Nebraska pheasant eyes.

57 rt: ...... •

4 hrs. 12 hrs. 16'12 hrs .

- , I I

,1\, ._­ �

20 hrs. 24 hrs. 28 hrs.

,9

32 hrs. 36 hrs. 44 hrs.

Color Plate No. 3 shows minimum physical changes that take place in pheasant eyes at various times after death at an ambient temperature of approximatcly .12°F . No corrections were made fo r wind and humidity. 4 hrs. 12 hrs. 16'/2 hrs.

20 hrs. 24 hrs. 28 hrs.

32 hrs. 36 hrs. 44 hrs.

Color Plate No. 4 depicts minimum physical changes noted in pheasant eyes at various times after death with an ambient temperature of approximately 40"F. No corrections were made for wind or humidity. 4 hrs. 12 hrs. 161f2 hrs.

20 hrs. 24 hrs . 28 hrs.

32 hrs. 36 hrs. 44 hrs.

Color Plate No. 5 illustrates mInIlTIUm physical changes that occur in pheasant eyes at various times after death at an ambient temperature of approximately 50°F. No corrections were made for wind or humidity . 2 hrs. 5 hrs. 10 hrs.

18 hrs. 22 hrs. 26 hrs.

2 hrs. 6 hrs. 10 hrs.

18 hrs. 20 hrs. 22 hrs. Color Plate No. 6 depicts minimum physical changes in pheasant eyes at various times after death at an ambient temperature of about 70°F. Two birds were used for these photographs, one for the first six and another for the last six . No corrections were made for wind or humidity . COTTONTAIL RABBITS (Sylvilagu s floridanus)

COTTONTAIL RABBIT SAMPLE COLLECTION ture was 500F, a rabbit with a body temperature of 700F died about 6 hours earlier. At OOF, the same rabbit would Iowa have died about 2.5 hours earlier. This would be 3.5 hours Rabbits and much of the data were supplied by coopera­ since death. Two rabbits together at 500F tend to insulate ting conservation officers. A special thanks to Cathy Hoilien one another and take an hour longer to cool to 700F than a for the long hours spent on the many aspects of this study. A single rabbit. At 500F a wet rabbit cooled to 700F one hour thank you also to Rosella Hallcock of the Iowa State Crime faster than a dry one (5 hours instead of 6 hours). When Lab and to professors Dr. David Roslien, Dr. Ken Abraham, ambient temperature fell to OOF, moisture appeared to have Dr. Rodger Knutson and Dr. Russel Rulon of Luther College little effect on cooling rate. Surrounding air temperature is at Decorah. Dale Anderson deserves thanks for his work on vital and obviously plays a major role in post-mortem cool­ the graphs. ing. In any report or study of this type, its'usefulness should TEMPERATURE be tested in the field . Iowa conservation officers have run Obviously there are many factors which influence body several hundred field tests under assorted conditions. They temperature at time of death. The most important of these noted variables such as ambient temperature, temperature would be surrounding temperature. Other factors would of storage sites, and moisture conditions. Laboratory tests include body weight, body covering, carcass handling, compared favorably with the field tests and for the most and certain weather conditions, especially humidity and part, were within a 5% error. For many rabbits, a working wind. Some of these factors are of more importance than range for TOD should be obtainable. The laboratory study others, but all several may have to be considered when indicated the accuracy involved in determining TOD if the making a TOD estimate. Since cottontails are predominately surrounding temperature was known and relatively constant. nocturnal, illegal taking after hours can be a common viola­ tion. In Nebraska many hunters will not take rabbits until their chances of contracting tularemia (rabbit fever) have ELECTRICAL STIMULUS decreased. This is in the colder months of the year as is As with other species, it was found that immediately hunting season. Many rabbits are probably taken after quail after death muscles react strongly. With the passage of time, or pheasant seasons open and by then many may be full­ the strength of this reaction slows but in varying degrees in grown. different parts of the body. Therefore, one can relate the Adult rabbits were used in the laboratory experiments length of time since death not only to the amount of re­ and were killed by a blow to the back of the head. A stand­ action to electrical stimulus but also to where on the body it ard Fahrenheit thermometer with a rubber band at 3 inches is found. insured accurate and uniform insertion into the rectum. Only the hair need be penetrated and skin touched. Temperatures were recorded every 15 minutes throughout The locations examined can be found in Figure 30. This the study period without fully removing the thermometer. technique appeared to be far less dependent on extenu­ The experiment included three controlled ambient temper­ ating circumstances, such as handling (dressing) of animal� or atures 500, 320 and OOF. At each temperature three carcass ambient temperatures (Table 21), than other techniques. conditions were tested. The conditions were: (1) a single Although fewer rabbits were examined than deer or rabbit, dry ,and intact; (2) two rabbits together, dry and geese, Table 21 along with Figures 30 and 31 should help in intact; and a single rabbit, wet and intact on a wet sur­ figuring ranges for time of death using electrical stimulus as a face. Both rab(3) bits in condition (2) were killed at approx­ guide. The muscle reaction to electrical stimulus provides a imately the same time and were placed in the same plastic method to estimate the TOD when an animal is fielddressed bag. or when conditions follOWing death are not clearly known. The decline in body temperature correlated to the sur­ The muscle reflex action is short-lived, about four hours for rounding air temperature (Figures 27-29). The TOD can be deer, three for rabbits and two for geese, raccoons, and determined from Figure 27. For example, if the air tempera- pheasants.

63 110°

:!: CD 90° .s::. c: Dry, unskinned , uncleaned CD .... 80° .s::.

If 70° c: ci

CDE .... ��-4--�-���--�--�� 50° >. "'0 Surrounding 0 �--+-��--+-��--+--F���-+--r-;-� m 32° Te mperature �--��-4--+--+--��--r-��--��--+--+--�-i Oo

o 2 3 4 5 6 7 8 9 10 II 12 13 14 15 16 Hours after death Figure 27. Post-mortem cooling of cottontail rabbits

Condition 1 - Single rabbit, dry , unskinned, uncleaned

Dry, unskinned t uncleaned .... 90° ·CD.s::. c: Two in a bag CD .... 80° .s::. If 70° c:

Q. SO°

CDE ..... r-4--+--��-+�r-�-+���-+--�4-�--+-� 500 >. 50° "'0 Surrounding 0 r-�-+--���--���r-�-+--��-4--+--+� m 40° 32° Te mperature 30°

20° 0 2 3 4 5 6 7 8 9 10 II 12 13 14 15 IS

Hours after death Figure 28. Post mortem cooling of cottontail rabbits

Condition 2 - Paired rabbits, dry, unskinned, uncleaned and laying together

64 110°

100°

.... "Ci 90° .s:::. t t c Wet unskinned uncleaned CD ... 800 .s:::. � 70° .-c

0. 60° E ....CD 50° >- "'0 50° 0 Surrounding m 40° 3 ° L---J-L-l---J-��--L--l--L-�-.l--l--L--=+���-J 2 Te mperature 30° 0°

20° 0 2 3 4 5 6 7 8 9 10 II 12 13 14 15 16 Hours after death Figure 29. Post-mortem cooling of cottontail rabbits

Condition 3 Single rabbit, unskinned, uncleaned and laying on a wet surface � D

Rabbit

ribs A J!::v

hind leg

\(c� e a r

tail

1/2 11/2 2 21/2 3

Figure 30. Locations on a rabbit where electrical Figure 31. Diagram depicting typical muscle response at four locations stimulus was checked. in cottontail rabbits.

65 Time Since Rectal Ambient Muscle Species Death (hrs) Temperature (oF) Temperature (OF) Reaction

Cottontail Rabbita 0-0.50 104 - 102 32 - 60 very good 0.51 - 1.00 102 - 100 60 very good 1.01 - 1.50 100 - 97 58 very good 1.51 - 2.00 97 -95 58 good 2.01 - 2.50 95 - 92 58 fair 2.51 - 3.00 92 -90 58 fair to poor 3.01 - 3.50 90 - 87 58 poor to none 3.51 - 4.00 87 - 85 58 none

Cottontail Rabbitb 0-0.50 104 - 100 32 - 100 very good 0.51 - 1.00 100 - 99 99 - 100 very good 1.01 - 1.50 99 - 98 96 - 100 very good 1.51 - 2.00 98 -96 96 good 2.01 - 2.50 96 -96 96 fair 2.51 - 3.00 96 -96 96 poor to none 3.01 -3.50 96 -96 96 none 3.51 - 4.00 96 -96 96 none

Cottontail RabbitC O. - 0.50 104 32 - 58 very good 0.50 - 1.00 58 very good 1.01 - 1.50 58 good to fair 1.51 - 2.00 58 fair to poor 2.01 - 2.50 58 poor 2.51 - 3.00 58 poor to none 3.01 - 3.50 58 none 3.51 - 4.00 58 none

Cottontail Rabbitd O. -0.50 30 very good 0.51 - 1.00 28 very good 1.01 - 1.50 28 - 26 fair 1.51 - 2.00 26 - 25 fair to poor 2.01 - 2.50 25 poor 2.51 - 3.00 25 poor to none 3.01 -3.50 25 none 3.51 - 4.00 25 none

aKilled with shotgun and placed intact in trunk, ambient temperature approximately 580 F. bKilled with shotgun and placed intact under heater in front seat of car; temperature ranged from 96-1OO oF. cKilledwith shotgun, fielddresse d, and placed in trunk of car, ambient temperature was 580F. dKilled with shotgun and placed intact in trunk ; ambient temperature was 25 300F.

Table 21. Post-mortem reaction to electrical stimulus in cottontail rabbits.

66 OTHER SPECIES

Data available for miscellaneous species has already been published or perhaps is not available in sufficient abundance to be given consideration for a separate section. It will be covered brieflyhere .

vania Game Commission and Penn State University Wildlife BLACK BEAR Unit personnel. (Ursus americanas) TEMPERATURE Temperatures were taken in the mouth and in the ear BLACK BEAR SAMPLE COLLECTION of these bears. Insertion was not forced but was normally as far as a thermometer would easily reach. Data is presented Pennsylvania in Figures 32 and 33 and represents bears weighing less than Temperatures of bears have been recorded when conven­ 100 pounds, to ones weighing 251 to 300 pounds. As could ient for several years. Information was taken at bear check be expected, the usual decreasing curves are illustrated. A stations and during live-trapping operations by the Pennsyl- more gentle decline was observed than was seen in pheasants.

BEAR MOUTH TEMPERATURE

110 6 LESS THAN 100 LB. 0 101 TO 150 LB. 0 151 TO 200 LB. 10 0 201 TO 250 LB. iii 251 TO 300 LB. 90 0 OVER 300 LB.

80

70 o C:. 0

60

&I. o 50 o

40 ------�- �7� I&JI It: ::l � 30 It: � � 20 ...

10

2 4 5 6 7 8 9 10 II 12 13 14 15 16 17 18 19 20

TIME HOURS AFTER DEATH

Figure 32. Post-mortem temperatures in the mouths of black bear with respect to time in hours.

67 BEA R EAR TEMPERATURE

.. LESS THAN 100 LB. • 101 - 150 LB. c::. 15\ - 200 LB. 110 o 201-250 LB. 251-300LB. 100

90

80

70

60

--- -__ ___ 27HR o

10

2 4 5 6 7 8 9 10 II 12 13 14 15 16 17 18 19 20

TIME HOUftS AFTEft DEATH

Figure 33. Post-mortem temperatures in the ears of black bear with respect to time in hours.

ELK taken for a period of up to 27 hours. Initial temperatures (Cervus canadensis) ranged from 100 to 1070 F rectally and 106 to 1120 F muscularly. Muscle temperature averaged 3 to 50 F higher ELK SAMPLE COLLECTION than rectal temperature at time of death. Increase in muscle temperature was attributed to chasing with a helicopter. Colorado The body temperature plotted against time since death As part of a physiological study, data was collected from revealed a curvi-linear relationship (Figure 34). In many car­ 48 elk, mean age of 5.8 years and mean weight of 248 casses a five hour plateau or an actual increase was observed pounds live, between June 1963 and April 1966. It was in elk muscle (ave. 2.40 F in 3.1 hours). Regression analysis supplied by Richard Denny, Harlan Riffel and Colorado displayed a simple linear relationship between elapsed time University's, Bob Mosley, Though not a primary objective, and carcass temperature. A strong relationship was found it was felt that temperature data might be useful to law between these factors (0.9332), Body temperature accounted enforcement personnel. for 87.1 % of the variance, body weight(0 .592) and ambient temperature were considered insignificant. The equation

TEMPERATURE derived fro:n Figure 34 was x = 40.3 -0 .3606y, where x =

During the course of this study, two elk were col­ time elapsed in hours since death and y = the body tempera­ lected each month from the White River and Rio Grande ture in degrees Fahrenheit. In 27 hours or less the equation elk herds. Samples were acquired either by euthanasia, yielded TOD estimates to within 7.3 hours with 95% confi­ with Cap-Chur darts from a helicopter, or shot on the ground dence limits. This technique may be useful in determining with a rifle. Both rectal and hind-quarter (3 to 4-inch inser­ whether an animal was possibly taken before or after legal tion, center of muscle mass by femur) measurements were hunting time.

68 , 122 , , , " • M 113 , " U , S , " C 104 " L " , E " • single temperature observation • 95 " o = multiple temperature observation T " " E " " " M " p 86 " , " • , E " R " 77 • A , T , • " U • • • " R 68 • • , E • " • • , , " • • of , . . . 59 ." , ... " . , • " . " , , 50 , " , '\. , • " , " 41 , " " '\." , 32 0 4 8 12 16 20 24 28 32 36 40 TIME IN HOURS FROM DEATH

Figure 34. Post-mortem muscle temperature in elk with respect to time in hours.

RACCOON ELECTRICAL STIMULUS (Procyon lotor) Electrical stimulus data is even more limited (Figure 36) but it appears that after about 2 hours, most reactions RACCOON SAMPLE COLLECTION have ceased. Reactions of the front foot, ear or eye will probably be the longest lasting. Reactions that are considered Nebraska very good have not been observed after an hour since TOD. Eight raccoon ranging in weight from 7.5 to 15 pounds were collected mainly from live traps or shot from trees. Cause of death was normally a .22 caliber bullet used to dispatch the raccoon. Ambient temperatures ranged from 30 to 700F, with the mean temperature being about 460 F. Temperatures were taken rectally using a 3-inch insertion. For electrical stimulus data, the side where the reaction was greatest was used, but damaged areas were avoided. Vitreous humor samples were difficult to obtain due to size and mobility of the eye. This data will not be presented here.

TEMPERATURE When taken rectally, live temperature appeared to average about 1020F. Figure 35 represents temperatures of dry (not wet) raccoons for a period, up to 25 hours after death. Over 180 temperature readings were made on these raccoons. It appears that for the first 10 hours one may be able to estimate TOD on dry raccoons to within a couple of hours. The line on Figure 35 is hand-drawn to show average temperatures. One would expect wet raccoons or those in the back of a pickup to cool faster, while larger raccoons and warmer weather raccoons, would cool somewhat slower. As Figure 36. Locations on a raccoon where electrical stimulus in all tests of this kind, judgment is required by the officer. tests may be conducted.

69 110

100

90

80 • � 0 0 � = �. ... o • e 70 Q) 0 Q., t � S Q) E-o � 60

• •

50 0

• • • 40

30 ' ______2 4 6 8 10 12 14 16 18 20 22 24 Post Mortem Interval In Hours

Figure 35. Post-mortem cooling curves for dry raccoons ranging from 7.5 to 15 pounds and at ambient temperatures varying from 40 to 60oF.

70 LITERA lURE CITED Adelson. L., I. Sunshine, N .B. Rushford and M. Mankoff. 1963. Vitreous potassium concentration as an indicator of the post­ mortem interval . J. Forensic Sci. 8 :503-5 14. Bate-Smith, E.C. and J .R. Bendall. 1956. Changes in muscle after death. Brit. Med. Bull. 12 :230-235. Beattie, K.H. and R.H. Giles, 1979. A survey of wildlife law enforcement research needs and current research. Wildl . Soc. Bull. 7:185-1 88. BendalL J .R. 1951. The shortening of rabbit muscles during rigor mortis: its relation to the breakdown of adenosine triphos­ phate and creatine phosphate and to musculer contraction. J. Physiol. 114:71-88. Bendall, J.R. 1960. Post mortem changes in muscle, p. 227-274. In G.H. Bournes. The structure and function of muscle.

Vol. 3: Pharmacology and disease. Academic Press, New York and London. Brisky , E.J ., R.N. Sayre and R.G. Cassens. 1962. Development and application of an apparatus for continuous measurement of muscle extensibility and elasticity before and during rigor mortis. J. Food Sci. 27:560-566. Brown, A. and T.K. Marshall. 1974. Body temperature as a means of estimating the time of death. Forensic Science, 4, 125-133 . Carleson, P and T.P. Kistner, 1982. Deer body temperature and time of kill. Interagency Memorandum, Oregon Dept. of Fish and Wildlife , Corvallis, OR. Coe , J.1. 1974. Post·mortem chemistry : practical considerations and a review of the literature. J. Forensic Sci. 18:12-32. Coe, J.1. 1972. Use ofchemical determinations on vitreous humor in forensic pathology. Forens. Sci. 17(4):541-546. Cowan, R.L., E.W. Hartsook. J.B. Whelan, J.L. Watkins, J.S. Lindzey, R.W. Wetzel and S.A. Liscinsky. 1968. Weigh your deer with a string. Pennsylvania Game News. 39(1 1): 17-1 9. Forster, B. 1963 . The contractible deformation of skeletal muscle in rigor mortis. 1. Forensic Med. 10:133-147. Ganong, W.F. 1981. Review of Medical Physiology, 10th Ed., Lange Medical Publications, Los Altos, CA. 628 pp. Gayet, A. 1900. The ocular signs of death. Gill, J.D. and D.C. O'Meara. 1965. Estimating time of eeath in white-tailed der. J. Wildlife Manage. 29(3):47 1-486. Hartman, F.E. 1977. Pheasant time since death study. Report from Pennsylvania Game Commission. 16pp. Hamilton-Patterson, J.L. and E.W. Johnson, 1940. Post-mortem glycolysis, J. Pathology and Bacteriology, 50(3):473-482. Hoilien, G.!. 1975. A study of time of death determination of cottontail rabbits by temperature. Iowa Conservation Com- mission, Law Enforcement Research. 9pp. Hoilien, GJ. 1976. A study of the variation of reaction to electrical stimulus after death. Iowa Conservation Commis- sion, Law Enforcement Research. 7pp. Hughes, W.M .H. 1965. Levels of potassium in the vitreous humor after death. Med. Sci. Law 5:15 0-156. Jaffe , F.A. 1962. Chemical post-mortem changes in the intra-ocular fluid. J. Forens Sci. 7 :231-237. Johnson, B.C., L.A. Maguire and D.R. Anderson. 1980. Determining time of death in mule deer by using potassium levels in the vitreous humor. Wildl. Soc. Bull. 8 :249-252. Karns, P.D. and K.D. Kerr 1980. Time of death estimate in moose . Proceedings of sixteenth North American moose work­ shop. 586 pp. Karns, P.D. and K.D. Kerr 1984a. Personal Communication. Minnesota DNR. Forest Wildlife PopUlations and Research Group, 2101 E. Hwy. 2, Grand Rapids, MN 55744. Karns, P.D. and K.D. Kerr 1984b. Time of death in white-tailed deer by analysis of vitreous humor potassium and glucose. To be published. Minnesota DNR, Forest Wildlife Populations and Research Group, 2101 E. Hwy. 2, Grand Rapids. MN 55 744. Keeton, W.T. 1969. Elements of Biological Science, W.W. Norton & Company, Inc., New York, NY, 582 pp. Kienzler, J.M. and W.A. Fuller. 1983. Estimation of time since death in white-tailed deer based on thigh temperature : A manual. Iowa Wildlife Research Bulletin No. 35. Iowa Conservation Commission. Des Moines, Iowa 39 pp. Kienzler, J.M ., P.F. Dahm, W.A. Fuller and A.F. Ritter. 1984. Temperature-based estimation for the time of death in \vhite­ tailed deer. Biometrics (in press). Kimball, C.F. 1972. Results from the Hunter Performance Survey 1970-1972 and the bag check temperature, 1972. Div. Wildl. Res. Migratory Bird and Habital Research Laboratory , Laurel, MD 20810,25 pp. Lowe, B. 1948. Factors affecting the palatability of poultry with emphasis on histological postmortem changes. Advances in Food Res. 1 :203-256. Lyle, H.P., K.L. Steammer, and F.P. Cleveland. 1959. Determination of the time of death; a consideration of postmortum physical changes. J. Forensic Sci. 4:167-175. Marburger, R.G. 1 966. To determine criteria for estimating time of death in white-tailed deer carcasses. Job completion report, Project W-62R, Edwards Plateau Game Management Survey in Texas. 3 pp. Marsh, B.B. 1952. Observations on rigor mortis in whale muscle. Biochim et. Biophys. Acta. 9:127-132. Moore, D.A. 1979. Five postmortem conditions compared to time of death in whitetail deer (Odocoileus virgin iallus) M.S. thesis. Northeast Louisiana Univ. Monroe, Louisiana, 80 pp. Morrison, P.R. and R.A. Hunt, 1963. Body cooling in dead waterfowl with special reference to estimation of the time of death. Department of Zoology and Wildlife Management, University of Wisconsin. Unpublished. Morrow, T.L. 1968. A literature review on postmortem changes, Colo. Coop. Res. Unit. Colo. State Univ .. Ft. Collins. Colo. 17pp.

71 Morrow, T.L. and F.A. Glover. 1967a. Experimental studies on postmortem changes in mallards. Bur. Sport Fish. and Wildl. Spec. Rept. Wildl. No. 134, 25pp. Morrow, T.L. and Glover F .A. 1967b Final Report : Determining time of death in mallards. Colorado Cooperative Wildlife Research Unit, Colorado State University, Ft. Collins, CO 80523 . pp. 42. Palmer, R.S. 1962. Handbook of North American Birds. Vol. 1. New Haven, Conn. 567 pp. Pex, J.O., K.D. Meneely and F.C. Andrews, 1983. Time of death estimation in blacktail deer by temperature and aqueous humor glucose. J. Forens. Sci. 28(3): 594·600. Pribor, H.C. and H.M. Bates, 1976. Clinical Forum, Lab . Mgt. 14(8):18. Prince , J.H. 1977. The eye and vision, Pages 696-7 12 in M.1. Swenson ed., Dukes physiology of domestic animals 9th ed. Comstock Publ. Assoc., Ithaca, N.Y. 914 pp. Rainy, H. 1869. On the cooling of dead bodies as indicating the length of time that has elapsed since death. Glasgow Medical Journal, new series 1,323-330. Reed, D.F. and D.C. Bowden. 1974. Postmortem thigh temperatures in mule deer. Outdoor Facts No. 98. Colo. Dept. Nat. Resour. 2 pp. Sarc.n,G. S.W. de. 1957. Estimation of the time of death by medical criteria. J. Forensic Med. 4:47-57. Schoning, P. and A.C. Strafuss, 1980. Determining time of death ofa dog by analyzing blood, cerebrospinal fluidand vitreous humor collected at postmortem. Am. J. Vet. Res. 41 :955-957. Schoning, P. and A.C. Strafuss, 1980. Postmortem biochemical changes in canine vitreous humor. J . Forens. Sci. 25(1): 53-59. Smart, C.W., R.H. Giles and D.C. Guynn. 1973. Weight tape for white-tailed deer in Virginia. J. Wildl. Manage. 37(4): 553-555. Sturner, W.Q. and G.E. Ganter. 1964. The postmortem interval. Amer. S. Clin. Path. 42: 137-144. Sturner, W.Q. and G.F. Gantner, 1964. Postmortem vitreous glucose determinations. J. Forens. Sci. 9(4): 485491. Van Den Oever, R. 1976. A review of the literature as to the present possibilities and limitations in estimating the time of death. Med. Sci. Law 16: 269-276. Woolf, A., J.L. Roseberry and 1. Will. 1983. Estimating time of death of deer in Illinois. Wildl. Soc. Bull. 11(1): 47-51. Woolf, A. and C. Gremillion - Smith. 1983. Some problems with using vitreous humor to determine time of death: problems and a review. Wildl. Soc. Bull. 11(1):52-55. Worcester, H.M. 1941. Preliminary report ofbi rd temperatures taken at Tule Lake Refuge, Unpublished. Wurdemann, H.V. 1920. The fundus of the eye after death. Amer. 1. Opthalmol. 3:321-323.

72