Lawson, Rommon L.

Lorenzen, Gerald A.

English

Northwestern University Illinois, School of Law: Scholarly Commons

Journal of Criminal Law and CriminologyVolume 62 | Issue 4 Article 171972A Possible New Approach for Determining thePost-Mortem IntervalGerald A. LorenzenRommon L. LawsonFollow this and additional works at: https://scholarlycommons.law.northwestern.edu/jclcPart of the Criminal Law Commons, Criminology Commons, and the Criminology and CriminalJustice CommonsThis Criminology is brought to you for free and open access by Northwestern University School of Law Scholarly Commons. It has been accepted forinclusion in Journal of Criminal Law and Criminology by an authorized editor of Northwestern University School of Law Scholarly Commons.Recommended CitationGerald A. Lorenzen, Rommon L. Lawson, A Possible New Approach for Determining the Post-Mortem Interval, 62 J. Crim. L.Criminology & Police Sci. 560 (1971)THE JOURNAL or CmninL LAW, CRIMINOLOGY AND POLICE SCIENCE Vol. 62, No. 4Copyright © 1971 by Northwestern University School of Law Printed in U.S.A.A POSSIBLE NEW APPROACH FOR DETERMININGTHE POST-MORTEM INTERVALGERALD A. LORENZEN ANrD ROMMON L. LAWSONGerald A. Lorenzen Ph.D. is a Research Biologist at the Engineering Experimental Station, TexasA&M University, College Station, Texas. After completing his graduate work at Texas A&M Uni-versity he served as an aerospace technologist for NASA and a research physiologist for the Depart-ment of Interior.Rommon L. Lawson, Ph.D. is in charge of the Radiation Testing Laboratory of the RadiationBiology Section of the Texas A&M Research Foundation, College Station, Texas. His research hasincluded the effects of radiation upon reproductive performance in various species.In many medico-legal cases there is a need toknow the approximate time of death. It has beenpointed out by Schleyer1 that the time of death isimportant in cases such as: unwitnessed fatal trafficaccidents, homicides, and some suicides. A solutionof a homicide case may depend upon a fairly ac-curate determination of the time of death, which isparticularly important when several successivecrimes have been committed by the same individ-ual. Therefore, it is essential that the determinationof the time elapsed since death be determined andsuch procedures justify extensive laboratory pro-cedures.A current review of the literature indicates atleast nine laboratory methods which show promisefor use in medico-legal cases. Biochemical methodsinclude the measurement of the variation in theamino nitrogen, non-protein nitrogen, creatine, am-monia, and inorganic phosphorus in the plasmaand/or the cerebrospinal fluid following death.Four physiological methods which have shownpromise are the measurement of the faradic excita-bility of muscles, the pupillary reactions upon in-jection of pilocarpine or homatropine into the an-terior chamber, the in vitro clotting of the fluidblood from peripheral veins, and the formation ofan idiomuscular contraction by mechanical irrita-tion. There are, of course, other methods whichare not considcred to bc laboratory techniques. Theabove methods are of no value after the thirtiethhour post-mortem, which in many cases offers thegreatest difficulty in assessing the time of death.According to Schleyer1 an exact chronology ofpost-mortem changes do not and cannot be ex-pected to exist. In all probability no single methodI Schleyer, Determination of the Time of Death in theEarly Post-mortem Interval, in 2 METHODS OF FORENSICScIENCE 253-93 (F. Lundquist ed. 1963).will ever provide a dependable range of accuracy,yet there can be little doubt that application of sev-eral tests may furnish the examiner with the degreeof certainty which is required for medico-legal de-terminations.When circulation ceases, tissue anoxia results innecrosis followed by decomposition. The time inwhich cells can continue to metabolize followingsomatic death varies according to the tissue andmay be erratic. Laboratory tests presently used todetermine the post-mortem interval depend uponthe time it takes for certain tissues to cease to func-tion, or upon chemical or physiological changes re-sulting from the necrosis. These tests in current useare only reliable up to the thirtieth hour post-mor-tem. Therefore, to make estimations of the post-mortem interval beyond the thirtieth hour, itbecomes necessary to look for some biological enti-ties which can live for extended periods of timewithout blood or oxygen supply.Ham2 reports that spermatozoa are anaerobicand can survive anaerobically on carbohydrates byfermentation. During fertilization they live for anumber of hours without oxygen, utilizing the car-bohydrates in the female genital tract for energy.According to Guyton, sperm can be stored andmaintain their fertility in the genital ducts for aslong as 42 days. Earlier reviews by Marshall4 statethat the spermatozoa of the bull may survive foras long as twelve days within the removed testicle ifthe latter be kept at a temperature a little abovethe freezing point. No documented informationconcerning the use of spermatozoa for estimatingthe post-mortem interval could be found.2A. W. HAm, HISTOLOGY 815 (1957).3 A. C. GUYION, TEXTBOOK OF MEDICAL PHYSIOLOGY1121 (1966).4 H. A. MARSHALL, PHYSIOLOGY OF REPRODUCTION177-78, 650.DETERMINING THE POST-MORTEM INTERVALA possible new approach for estimating the post-mortem interval beyond the thirtieth hour is sug-gested by sperm survival in the genital tract. Apreliminary investigation was, therefore, conductedto determine the concept that sperm viability afterdeath may be utilized for estimating the post-mor-tem interval.METHODSTwo sets of bovine testes were made available tous by the local slaughter houses. The scrotum andvas deferens had been removed leaving the testesand epididymes enclosed in the tunica. The head,body, and tail of the epididymes were completelyintact.One set of testes had been in storage for severaldays (168 hours) and the other set had been re-moved from a bull which had been dead only twohours. Both sets had been maintained at an averagetemperature of 38*F after slaughter. The testeswere placed in plastic bags containing a smallamount of normal saline and stored in the labora-tory at 38°F. Sperm samples were immediatelytaken from each specimen by making a- small slitin the wall of the epididymis. A sample was takenof the epididymal fluid which extruded from theincision onto a dean glass slide. Two slides wereprepared for each specimen. The epididymal fluidon one slide was mixed with a drop of egg yokesemen extender- which had been stored at the sametemperature as the specimens, since changes intemperature result in a high mortality rate to thesperm. A cover slip was placed over the preparationto prevent drying and the motility of the sperm es-timated. The specimen for the second slide wascollected in a similar manner and prepared accord-ing to Sorensen and Miller.6 A drop of Schaffer'sand Almquist's7 differential stain was added to theepididymal fluid and thoroughly mixed. A thinsmear was prepared by placing another dean slideon it and drawing it along with a gentle motion.The slide was then dried over the flame of a Bunsenburner. The differential stain is a staining mixtureof 1% eosin B and 4% aniline blue dissolved in M/8phosphate buffer having a pH of 7.2. This stain isused to differentiate between live and dead sperm;the heads of dead sperm are stained light blue,while live sperm remain unstained. "I E. J. PERRY, THE AReTiFIcrAL INsE~MINATION OFFAXm ANnEms 116 (1952).6 A. M. SORENSEN & L. M1i.ER, AN iAL REPRODUc-TION, A LABORATORY MANUAL.7 Shaffer & Almquist, Relation of the Eosin-AniineBlue Staining Method to the Quality of Bull Semien, 32J. DAmY Scr. 723 (1949).On the following day one epididymis from eachset was placed in a constant temperature oven at90°F. One specimen was 24 hours old, and the otherwas 192 hours old. Microscopic observations werecontinued every twenty-four hours through thefourth day on the refrigerated samples. After 24hours, the tissue decomposition of the incubatedspecimens became so advanced that samples couldno longer be taken. It should be noted that theseven day old sample was manifesting signs of de-composition before it was incubated. At the termi-nation of the study the older refrigerated samplewas 288 hours (12 days) old.RESULTSInitial microscopic observations showed numer-ous motile sperm in both the 168 hours and 2 hour-old specimens. The sperm manifested the samenormal pattern of motility as seen in a healthybovine semen sample.According to the differential staining technique,which distinguishes between live and dead sperm,approximately 80% of the sperm in both the 168and 2 hour-old refrigerated specimens were alive.This percentage slowly declined in the older speci-men to an average of forty percent at 240 hours.At the final observation after 288 hours about 50%of the sperm were fully stained and the remaining50% were partially stained. This would indicatethat 50% of the sperm had died recently and thecell membrane was in the initial stages of becomingpermeable to the dye.There was a steady decline in viable sperm in the2-hour old refrigerated specimen. According to theobserved motility and staining results it was esti-mated that about 50% of the sperm were still aliveat 120 hours post-mortem. The extent of decom-position made it impossible to get representativesamples from the incubated specimens after 24hours. Both incubated specimens exhibited viablesperm when sampled at 24 hours but percentageestimates were not made.The sperm appeared normal in all slides studied.Figure 1 shows the sperm in the samples taken at48 hours following a 24-hour incubation period at90°F, and figure 2 shows a representative sampletaken at 288 hours at 38F, respectively. Numerouswell-formed spermatozoa were present in all slidesstudied.DIscUssioNThe results of this study show that bull sperma-tozoa, and presumably those of other mammals in-1971]t" I I I I i -2RSperitoo fr t X lhour-, at 9%Sperratozo taken from and n a 2i Rw Ii4 Mm a adrcluding the hniy ni lr eai viaikdciymhsfo aneteddeioofthe tThepost-r v iture d n ieased teperaturecre:se T1 etal)oAc rat[e of. spe I rmLshortn lic spa. It wila,-S of the surrodi i)zoag, an'dsue, Thccomibnain of those two factrs cot ibule o theincreasedc dleat,,h ratmight bc diree ' an'i, tkrval,c of (he spermatozoa whichindicator of the post-nortemUhis Prelim r I studly has provoked interest inedrfig e a of uthicng spermatozoa orMeated tra oJ es ating the interval afterdeath lbeyond thbty mors, Possibly some of t:heot~her methods whichi have been udsed f~or estimatingUT 11x Y~u-. ' peiod of 24DETERINING THE POST-MORTE:.M INTER V.A Lthe quality of semen in animal science could beutilized for estimating the age of the sperm follow-ing death, and, correlated with the ambient con-ditions, give a dependable method of estimating thetime following death of the individual.These methods measure (1) the duration of sper-matozoa motility at a standard low temperature,(2) the glycolytic power of spermatozoa, (3) oxygenconsumption of spermatozoa and, (4) the resistanceof spermatozoa to temperature shock or a 1 %sodium chloride solution. Perhaps a modification ofthe Methylene Blue Reduction Technique of Beckand Salisbury which gives an index of the concen-tration and activity of the sperm could be usedhere.Methods should be developed for correlating theviability, motility, death rate and perhaps morpho-logical and histochemical changes in the epididymalfluid with time and temperature.In its simplest form this method could be used toroughly estimate the post-mortem interval, if thelength of time that epididymal sperm will remainalive or motile under a certain set of conditionswere known. For example, if it were known thatsperm are motile for 10 days in the epididymis of abody which has remained at a temperature varyingbetween 320 and 38'F following known time ofdeath, it would be possible to estimate whether abody in which the time of death was unknown hadbeen (lead greater than or less than ten days.If this method proves feasible it would probablyhave its greatest value in the colder climates orduring the winter. Much more research is requiredto establish relationships of viability of sperma-tozoa with temperature and other ambient condi-tions. The results presented in this paper indicatethat sperm remains viable and motile during apost-mortem interval where existing tests are un-reliable.19711

A Possible New Approach for Determining the Post-Mortem Interval

bepress Legal Repository

https://scholarlycommons.law.northwestern.edu/cgi/viewcontent.cgi?article=5758&amp;context=jclc

A Possible New Approach for Determining the Post-Mortem Interval

Abstract

Similar works

Full text

Available Versions

Northwestern University Illinois, School of Law: Scholarly Commons

bepress Legal Repository