University of Montana ScholarWorks at University of Montana

Graduate Student Theses, Dissertations, & Professional Papers Graduate School

2000

Forensic entomological case study and comparison of burned and unburned Sus scrofa specimens in the biogeoclimatic zone of northwestern Montana

Seth Barnes The University of Montana

Follow this and additional works at: https://scholarworks.umt.edu/etd Let us know how access to this document benefits ou.y

Recommended Citation Barnes, Seth, "Forensic entomological case study and comparison of burned and unburned Sus scrofa specimens in the biogeoclimatic zone of northwestern Montana" (2000). Graduate Student Theses, Dissertations, & Professional Papers. 4655. https://scholarworks.umt.edu/etd/4655

This Thesis is brought to you for free and open access by the Graduate School at ScholarWorks at University of Montana. It has been accepted for inclusion in Graduate Student Theses, Dissertations, & Professional Papers by an authorized administrator of ScholarWorks at University of Montana. For more information, please contact [email protected]. , Maureen and Mike MANSFIELD LIBRARY

The University fMONTANAo

Permission is granted by the author to reproduce this material in its entirety, provided that this material is used for scholarly purposes and is properly cited in published works and reports.

** Please check "Yes" or "No" and provide signature * *

Yes, I grant permission No, I do not grant permission

Author’s Signature

Date____

Any copying for commercial purposes or financial gain may be undertaken only with the author's explicit consent. Forensic Entomological Case Study and Comparison of Burned and Unburned Sus Scrofa Specimens in the Biogeoclimatic Zone of Northwestern Montana

by

Seth Bames

B.S., University of Wisconsin-Oshkosh

Presented in partial fulfillment of the requirements

for the degree of

Master of Arts

The University of Montana-Missoula

2000

Approved by:

£ Chairman

Dean, Graduate School

, S" (*?" 2ooo Date UMI Number: EP40119

All rights reserved

INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted.

in the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion.

UMI* Dissertated rtifcfeMng

UMI EP40119 Published by ProQuest LLC (2014). Copyright in the Dissertation held by the Author. Microform Edition © ProQuest LLC. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code

ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106-1346 Barnes, Seth, M.A., Spring 2000 Anthropology

Forensic Entomological Case Study and Comparison of Burned and Unburned Sus Scrofa Specimens in the Biogeoclimatic Zone of Northwestern Montana (75 pp.).

Chairperson: Randall Skelton The goal of this experimental case study is to provide baseline rural forensic entomological data for the Interior Douglas-fir/Ponderosa Pine biogeoclimatic zone of Missoula County in Western Montana. The first hypothesis is that successional fauna will be similar to that of Dillon (1997) who conducted well-replicated forensic entomological studies in a similar biogeoclimatic zone located in British Columbia, Canada. The second hypothesis states that the burned and unbumed specimens will have different successional fauna specific to their physical states. The third hypothesis is that the insect successional pattern and species diversity on carcasses present in a mixed urban/rural and rural area will be distinct (Ternenyl997). This will provide the forensic investigator with a species diversity index indicating the abundance of urban and rural species in two biogeoclimatic zones of Montana. The first pig was burned with one gallon of gasoline and the second pig remained unbumt to act as the control. Phormia regina (Meigan) adults and immatures dominated among the order Diptera throughout the study until the post-decay stage when they co­ dominated with Lucilia illustris (Meigen). Protophormia terraenovae (Robineau- Desvoidy) also played a small role. During the post-decay stage Prochyliza brericornis (Melander) and Prochyliza xanthostoma (Walker) of the family Piophilidae made a significant contribution to the decomposition process. Among the Coleoptera Thanatophilus lapponicus (Herbst) and Necrodes surinamensis (Fabricius) were the most representative families. Many similarities were found between this study and that of Dillon (1997) in a similar biogeoclimatic zone. The parallels in decay rates and species diversity for these two studies conducted in an interior Douglas-fir and Douglas-fir/Ponderosa Pine Rocky Mountain biogeoclimatic zone cannot be accounted for as coincidence. The insect successional pattern was very similar for both pigs. The burned pig showed only a slight difference in the rate of decay and successional pattern. The families of insects involved in the decomposition process were similar for both the urban/rural and rural areas. The insects were typed to genus or species in this study. If applicable to human decedents, this baseline data will aid in the interpretation of Montana forensic anthropology and autopsy results to provide the medico-legal community with a more lucid explanation concerning time since death in forensic cases. Acknowledgements

I would like to thank all who made this study possible. I would like to thank Frank, Patty, and Chris of the Lubrecht Experimental Forest for the use of equipment, housing, and their generous help. I would also like to thank Diana Six of the School of Forestry, University of Montana, for use of her laboratory, equipment, and help in identification. Additional thanks to Douglas Emlen of the Division of Biological Sciences, University of Montana, for his advice and assistance. Extra thanks to Elena Ulev for assistance in completing fieldwork and to my committee members for guidance. Finally, a thank you to everyone who helped in their own w ay, including the two Sus scrofa who became test specimens. Table of Contents

Title Page Abstract Acknowledgments Table of Contents List of Tables List of Figures Chapter 1-Introduction Chapter 2-Materials and Methods Chapter 3-Results Chapter 4- Discussion Chapter 5-Conclusions Tables Figures References List of Tables Table Number Page Number

1. Insect Succession for the burnt Sus scrofa------46-49

2. Insect Succession for the control S. scrofa------50-53

3. Decay stage lengths from Dillon (1997) and the present study ------54

4. Decay stage lengths from Temeny (1997) and the present study ------54 List of Figures

Figure Number Page Number

1. Diptera Species Diversity, Fresh: Burnt and Control S. scrofa ------55

2. Diptera Species Diversity, Bloated: Burnt and Control S. scrofa------56

3. Diptera Species Diversity, Decay: Burnt and Control S. scrofa------57

4. Diptera Species Diversity, Post-Decay: Burnt and Control S. scrofa------58

5. Diptera Species Diversity, Remains: Burnt and Control S. scrofa------59

6. Coleoptera Species Diversity, Bloated: Burnt and Control S. scrofa------60

7. Coleoptera Species Diversity, Decay: Burnt and Control S. scrofa------61

8. Coleoptera Species Diversity, Post-Decay: Burnt and Control S. scrofa 62

9. Coleoptera Species Diversity, Remains: Burnt and Control S. scrofa------63

10. Ambient and Internal Temperatures, days 1-30: Burnt and Control S. scrofa—64 Introduction: Chapter 1

The benefits and uses of forensic entomology are many. Insects have been used in forensic science on nearly every continent in a number of diverse climates. This variety of biogeoclimatic zones ensures that comparisons of data will show striking contrasts as well as remarkable similarities. The widespread application of forensic entomology is not surprising because it is typically used for conducting criminal investigations. Every year forensic entomology plays an increasing important role in the pursuit of justice.

The characteristic role of the forensic sciences is to provide law enforcement agencies with comprehensive examinations of evidence provided by law enforcement personnel. This evidence is commonly examined by crime laboratory sections such as serology and latent prints. Unfortunately these sections rarely, if ever, examine entomological evidence. Only the coroner or medical examiner frequently have the opportunity to inspect such evidence, and in most cases, these individuals do not have an extensive background in the study of insects. At this point, the entomologist is able to contribute their scientific observations to the overall examinations of case evidence. This data may provide law enforcement officials with the information they need to successfully bring closure to open cases. Several areas of entomology may serve this end.

Lord and Stevenson (1986) recognize three areas of forensic entomology: urban, stored-product, and medicolegal. Urban and stored-product investigations usually involve lawsuits pertaining to civil and consumer domains, respectively. Medicolegal forensic entomology examines arthropod association with illegal activities, such as

1 murder. Utilization of this type of entomology involves determining the time since death, or Post Mortem Interval (PMI). The PMI may be estimated in two ways. The first method is to estimate the development time of each species collected at the scene. This estimate is primarily used in the early stages of decomposition. Ordinarily those insects exhibiting the longest developmental period of growth are used to ascertain the PMI.

The second method involves corpses in an advanced stage of decomposition.

This method involves observing arthropod successional patterns and compares them with preexisting data for the same or similar arthropods in the relevant biogeoclimatic zone in question. These methods complement examinations made by other forensic investigators, including forensic anthropologists.

Both methods require the determination of behavior of the arthropods present.

Goff (1993) presents four types of arthropod feeding behavior, all of which may be found during forensic investigaitons. The first type of behavior is necrophagy, feeding directly upon the corpse. The second types of behavior is predation and parasitism. This group frequently consists of individuals in the orders Coleoptera, Hymenoptera, and Diptera, and frequently affect eggs and larva. The third type of behavior is omnivory, feeding upon both corpses and other arthropods present. The fourth type of species is adventive with use of the corpse possible in many different ways. Some of these species may feed upon mold or fungi growing on the corpse while others simply extend their normal habitat to include the microenvironment provided by the corpse.

Forensic entomology and anthropology share an interest in the circumstances and processes surrounding a corpses process of decay. The study and application of forensic entomology has, until recently, been relatively uncommon in the United States of

2 America. The only institution legally sanctioned to use human corpses is the

Anthropological Research Facility (ARF) in Tennessee. However, most researchers frequently use animal models for surrogate humans. At ARF, corpeses are allowed to decompose under a variety of conditions to simulate actual medicolegal forensic cases.

Animal models have been used in temperate environments (Dicke and Eastwood 1952,

Reed 1958, Payne 1965, Johnson 1975, Baumgartner 1988, Blackith and Blackith 1990,

Dillon 1997, Temeny 1997, and Adair 1999), tropical environments (Early and Goff

1986, Goff et. al. 1986, Goff and Odom 1986 and 1987, Goff 1992b) and arid environments (Baumgartner 1986, Galloway et. al. 1989).

Entomological investigations are common outside the United States. A few of the earliest works in Europe include those by Redi (1668), Bergeret (1855), Brouardel

(1879), Megnin (1888, 1894), and Motter (1898). Additional case studies and experiments from the twentieth century include those conducted in such diverse countries as Finland (Nuorteva et. al. 1967 and Nuorteva 1970,1974,1977 and 1987), Italy

(Introna et. al. 1998), Australia (Palmer 1980), South Korea (Rueda et. al. 1997), Egypt

(Tantawi et. al. 1996 and 1997), and Japan (Nishida 1984). Many investigations are not listed here but may be found in sources such as Meek et al (1983), Vincent et al (1985) and Smith (1986).

One of the earliest works completed by Redi (1668) consisted of leaving meat exposed to, or protected from, flies. This work showed that spontaneous generation of flies did not occur, and that flies contributed to the decay process. Bergeret's (1855) later medicolegal forensic entomology work provided insight into similar methods and materials still used in today's investigations. But it was Megnin's work from 1883-1898

3 which suggested for the first time that the process of decomposition occurred in a predictable manner. These early studies gave status to the budding branch of entomology among the forensic sciences. More recent and broad examinations of the subject have been published by Smith (1986) and Catts and Haskell (1990).

The detection of various pharmaceuticals in forensic cases has also received treatment in recent studies. Goff et. al. (1989,1991, 1993 and 1994) have examined the evidence of the effects of cocaine, heroine, and other various chemicals upon decaying tissues and the arthropods associated with them. Goff and Lord (1994) refer to this new field as entomotoxicology and believe it has good prospects of becoming a standard forensic technique when insects and chemicals are found together.

Forensic anthropology is similar to forensic entomology in that both attempt to ascertain data about the deceased, frequently with only a small amount of material to work with. The main contrast is that forensic anthropologists focus upon the bones of the individual to determine aspects such as age, stature/weight, paleopathology, and sex. It is difficult for the anthropologist to determine how long the individual has been dead, apart from cultural markers. This is where entomology contributes as a partner in solving a forensic case. With the entomologist's estimate of time since death (PMI), the information gathered by the entomologist and anthropologist may be combined to form a comprehensive evaluation of the activities surrounding the individual before and after death. Until now the only forenstic entomological study from Montana was conducted by

Temeny (1997). Apart from several forensic cases (Barnes, unpublished data), forensic entomology has not been used extensively within Montana.

4 Within the realm of forensic entomology one may discern:

• Whether the body was moved after death. Many insects are limited to particular environments and are rarely found elsewhere (Goff 1992a). For example, aquatic insects found on a corpse in an arid environment would indicate that the deceased spent some amount of time in an aquatic environment after death.

• Whether the individual was killed inside or outside a structure (Meek, unpublished,

Haskell, unpublished). If insects typically found in urban environments, such as those feeding upon stored goods, are found upon a corpse in a rural environment, it could be deduced that the individual was killed inside a building and then transported away from the urban environment.

• Whether the individual was killed during the day or the night (Nuorteva 1977, Smith

1986 and Benecke 1996). Species of insects have distinct diurnal or nocturnal activity patterns.

• The presence of antemortem drugs (Catts and Goff 1992b) by conducting tests upon larvae.

• The possible effect of insect activity on mimicking sexual assault clothing patterns

(Komar and Beattie 1998).

• Patterns of insect succession in wildlife death investigations (Dillon 1997).

• Evidence of child abuse or neglect (Goff et.al. 1991 and Lord 1990).

• Evidence of insect interaction with a crime suspect (Webb et.al. 1983 and Prichard et.al. 1986). Suspect may have insect bites or insects on their person which are consistent with entomological evidence found at the crime scene.

5 Erzinclioglu (1985) stated that each forensic case must be treated as unique and that no basic rules of forensic entomology can be formulated. Yet, baseline data are required in each biogeoclimatic zone if forensic entomology cases are to produce positive and useful information. The intention of this study is to confirm baseline data previously collected in the northern region of the Rocky Mountains in a similar biogeoclimatic zone

(Dillon 1997). If the fauna are similar, a forensic investigator maybe able to extrapolate

Dillon's data to forensic cases in a similar biogeoclimatic zone in Montana and vice versa. A biogeoclimatic zone may be defined as a geographic region which has a distinctive climate and supports organisms specific to that region when compared with bordering regions or zones.

Data from this study Will also be compared with the only other "bum study" published in forensic entomology to this date (Avila and Goff 1998). The burned specimen is meant to simulate an accidental death or homicide victim burned with an accelerant. The standards for the extent of the bum have been set by Glassman and Crow

(1996) who divide the identification of fire injury to an individual into five levels. The final aim of the project is to complement the only existing forensic entomological study conducted in Montana, which was performed in the same biogeoclimatic zone at a lower elevation in a mixed urban/rural setting (Temeny 1997). This will provide the forensic investigator with a species diversity index indicating the abundance of urban and rural species in the interior Douglas-fir/ponderosa pine biogeoclimatic zone of Montana.

Results of this study may also be valuable in determining if the decedent was physically moved after death and possibly by whom. These three hypothesis may be stated as:

6 (1) The successional fauna of the pig model will be similar to that found by Dillon

(1997) who conducted well replicated forensic entomological studies in a similar biogeoclimatic zone located in British Columbia, Canada.

(2) The burned and unbumed specimens will have different successional fauna specific to their physical states.

(3) The insect successional pattern and species diversity on carcasses present in a mixed urban/rural and rural area will be distinct (Temeny 1997).

It has been established that pigs, being omnivores, have similar gut fauna as well as roughly an equal amount of body hair as humans. Accordingly, the use of pigs as surrogate humans in decay studies is well established (Payne 1965, Payne et. al. 1968,

Payne and King 1970 and 1972, Smith 1986, Tullis and Goff 1987, Haskell 1990,

Hewadikaram and Goff 1991, Shean et. al. 1993, Dillon 1997, Temeny 1997). Therefore, the animal model chosen for this experiment was the common pig, Sos scrofa.

While pigs are viewed as suitable animals for forensic studies, the size of a pig can have important effects on insect colonization. Previous experiments have used animal models ranging from a small size of approximately 25 kilograms or less (Hackman 1963,

Comaby 1974, Blackith and Blackith 1989 and Patrican and Vaidyanathan 1995), a medium size of approximately 50 kilograms (Fuller 1934, Bomemissza 1957, Reed 1958,

Burger 1965, Johnson 1975, Jiron and Cartin 1981, Early and Goff 1986, Tantawi 1996,

LeClercq 1997), to a relatively large animal model, such as an elephant (Coe 1978 and

Braack 1986). Arguments for or against a particular size commonly revolve around decay rate as opposed to insect succession or species.

7 In a study pertaining to effects of specimen size, Hewadikaram and Goff (1991) concluded that successional patterns and developmental rates of insects were not dependent upon specimen size. Their largest pig was twice the size of the smallest and attracted a greater number of taxa, with a marked difference between days 5-16 of the study. This attraction of a greater number of taxa is probably related to increased surface area and mass available for oviposition. The authors found no difference between carcass size and the rate of development for arthropods, which are used for determining the PMI.

They deduced that arthropod succession patterns and developmental rates are independent of the size of the animal model. However, Komar and Beattie (1998) used pigs ranging in size from 19-162 kilograms and determined that larger carcasses attract a greater number of arthropods and support larger maggot masses than do small carcasses.

Therefore, while development and succession may not differ in pig carcasses of differing size, relative abundance of insects may.

8 Chapter 2: Materials and Methods

Study Area

The study site was located in northwestern Montana (46’53'N, 113'27'E) at the

Lubrecht Experimental Forest (LEF) in Missoula County. Lubrecht is approximately 30 miles (53 kilometers) east of Missoula, MT. The forest comprises 28,000 acres (10,927 ha) of zoned and managed areas with uses ranging from recreational to resource conservation and range management. The study site is found within an area zoned for grazing, hunting and nature study (Maus, personal communication). It is in range and township of R15W T13N on the northern border of section 15. The site is located at the end of an unimproved secondary road, located 1 mile southwest of the Lubrecht

Experimental Forest headquarters off Highway 200. The road proceeds northwest of

Highway 200 for approximately 2 miles. The elevation of the study site is nearly 4,400 feet and is located on the southward facing slope.

Vegetation consists predominantly of coniferous species in low to middle elevation categories (Hitchcock and Cronquist 1973; Eyre 1980). The primary forest cover is a mix of ponderosa pine (Pinus ponderosa) and Douglas-fir ( Pseudotsuga menziesii). Understory vegetation consists of snowberry ( Symphoricarpos duhamel), ninebark (Physocarpus maxim.), various grasses, and other vegetation common to these stands.

Animal Cadavers

Two pigs were used in this study and therefore observations were not replicated.

Replication is preferable in experiments of this nature. Therefore, this project should be

9 viewed more as a long-term case study with the intention of gathering baseline data for the biogeoclimatic zone of northwestern Montana.

Two mature 180-pound (81.65-kilogram) pigs were selected for the animal model in this study. Each was killed with a single gunshot to the head, producing only a small entrance wound. Both pigs were placed in separate bear proof cages measuring 150 centimeters in length by 90 centimeters in height by 90 centimeters in width and constructed with re-bar steel. The control pig (CSS) and cage were placed 30 meters from the pig to be burned (BSS).

Once in the cage, one pig was sprinkled with 3.8 liters (1 gallon) of gasoline and set afire. After the pig had been allowed to bum for approximately 30 minutes, flames began to wane and small cracks appeared upon the skin surface with a small amount of charring

(Figure 1). Once the flames dissipated, the level of bum apparent on the pig was evaluated using the Crow-Glassman scale (Glassman and Crow 1996).

Climatology

The experiment began on June 17, 1999. Temperatures were measured using a hand­ held thermometer at the following locations: internal temperature (approximately 3 inches under the skin), the skin surface, the ground surface and between the specimen and the ground surface in the area of the abdomen. Measurements were taken twice a day, once in mid-morning and once again in mid-afternoon for the first 30 days of decomposition, then similarly every two days for a period of approximately 60 days, and finally one day a week until December 1st, 1999, when travel to the site was no longer possible due to snow. Ambient local site temperatures and humidity were recorded continuously from June 17,1999 to December 1,1999 using a hygrothermograph.

10 Rainfall was recorded using a standard rain-gauge located at the site and monitored at the same time as the hand-held readings discussed above. Local cloud cover conditions were noted visually. All ambient temperature and rain measurements were compared with information from the nearest National Weather Service (NWS) station located at the

Missoula County Airport in Missoula, MT.

Decomposition Stages

The framework selected for this experiment was designed by Early and Goff

(1986) and reviewed by Goff in 1993. The classification system used consists of five stages, which indicate arthropod successional patterns, chemical decay processes, and physical transformations in the carcass or corpse. Transitions from one stage to the next are continuous. The five stages are summarized briefly as follows:

1. Fresh Stage

This stage begins at death and ends when bloating is visible. The first arthropods

to colonize the specimen usually consist of the families Calliphoridae (blowflies) and

Sarcophagidae (flesh flies). Abundance and species diversity during this stage is

dependent upon the biogeoclimatic zone and a host of environmental variables. Flies

present at this stage will deposit eggs or larvae on natural orifices or open wounds of

the specimen.

2. Bloated Stage

At this stage, anaerobic bacterial action produces gasses that inflate the carcass.

The feeding activity of dipteran larvae and putrefaction increase the carcasses

temperature. The combination of carcass fluid seepage-and the ammonia of dipteran

11 larval activity cause the soil under, and immediately surrounding, the carcass to

become more alkaline, thus changing the faunal content of the soil.

3. Decay Stage

Gasses begin to be released and the carcass ceases to be bloated. The large

number of larvae present increase the presence of predaceous arthropods. The end of

this stage is indicated when the majority of dipteran larvae have migrated away and

the carcass has been mostly consumed.

4. Post-Decay Stage

This stage finds a transition of the carcass into primarily skin, connective tissue,

and bones. Coleoptera as well as predatory and parasitic species increase in number.

5. Remains Stage

The only portions of the carcass remaining at this stage are bones and hair. Soil samples indicate a return to normal fauna. The length of this stage is extremely variable depending upon the biogeoclimatic zone. Times may vary from years in an extremely dry environment to months in a tropical environment with high scavenging activity.

Insect Sampling

Insects were collected only from the external surface of the pigs to avoid altering conditions of the cadavers. Detailed notes were taken of where the insects were located on the body according to procedures outlined by Lord and Burger (1983) and Borror et. al. (1989). Photographs were taken each day to illustrate locations of insects and the status of the carcasses. Samples were collected from each carcass daily during the first

30 days of the experiment. Once into the decay stage, samples were collected once every

3 days until the post decay stage was apparent. Sampling was decreased to once every 7

12 days when the post decay stage occurred. This rate of sampling continued until snow and low temperatures hindered arthropod activity as well as collection procedures.

Collection procedures for each decomposition stage were limited to the mode of locomotion of the arthropods present. During the bloated and decay stages many fly larvae and adults were collected. The procedures for the collection and storage of these different stages of Diptera development are distinct. Furthermore, the collection techniques are also different depending upon the size of the Diptera being captured.

These procedures and techniques are detailed below.

Insects: Collection Techniques

Adult Insects

This experiment used procedures basic for both ground-dwelling as well as flying specimens. Adult ground-dwelling insects were collected in two ways. Pitfall traps were used to capture adults as well as migrating larvae. Each pitfall trap consisted of a medium sized glass jar approximately 15 centimeters (6 inches) tall and 7.5 centimeters

(3 inches) wide at the mouth. A piece of metal screening (.625 centimeter or 1/4 inch mesh) material measuring 2.5 centimeters by 2.5 centimeters (1 inch by 1 inch) was fitted over the mouth of the jar. Soapy water was placed in the bottom of the jar to ensure insects could not escape. A single pitfall trap was placed approximately 30 centimeters

(1 foot) from each carcass inside the cage to prevent small mammals being captured or injured. Forceps and fingers were also used to capture many ground-dwelling insects.

A net was utilized for large flying insects such as Calliphoridae and

Sarcophagidae. Two-dozen sweeps were made each day over each cadaver to capture a representative sample of insects. An aspirator was used for smaller specimens such as

13 Phoridae and Piophilidae. Captured specimens were killed in ethyl acetate killing jars and then transferred to vials containing 70% ethyl alcohol. Labels for vials included the site name, carcass type, county, state, date, time, sample number, and location of insect.

Larval Insects

Collection of larvae was accomplished using forceps. Samples were taken every day larvae were present, with a small portion of each sample saved for rearing purposes.

Portions not used for rearing were placed in vials containing Pampel's solution, a preservative that helps retain the larvae's size and natural coloration (Smith 1986).

Larvae migrating from the carcasses were collected and labeled separately from those collected on the cadavers with migratory distances noted. Migration routes and distances are important in finding specimens at a death scene, therefore, empty pupal cases were also collected from near the cadavers.

Pupal Collections

The use of pupae in the forensic sciences has not been extensive (Malloch 1917,

Gilbert and Bass 1967, Okely 1974, Chu and Wang 1975 and Nuorteva 1987). Several authors have published repports indicating that the age determination of pupae is significant to forensic investigations (Reiter and Wollenek 1983, Nuorteva 1987,

Greenberg 1988 and Catts 1991). Once the age of the pupae has been established, it lends credence to the estimation of the PMI. Therefore, the discovery and collection of pupae is an important part of the overall forensic entomological data gathered at a crime scene. Archaeological methods were employed here in the hope that using a specific technique of sampling and excavation would yield better results than using non­ archaeology techniques.

14 The migration of larvae from a carcass occurs at varying times and ways depending upon the insect taxon and stage. The last larval stage often pupates in the soil directly under or adjacent to their food source. However, some larvae often "wander"

(between 3 and 10 meters) from a carcass to avoid competition and to seek dry soil for pupation (Smith 1986). After emergence, the empty pupal case remains.

Haskell and Williams (1990) found that samples should be collected up to one meter from the body for good coverage. If maggots have left the corpse it is important to track down and capture puparia, inactive larvae, and prepupae. Smith (1986) reccommends making shallow 7 centimeter (3 inch) transects in several compass directions of up to 20 feet from the corpse.

Puparia were sampled using systematic sampling (Renfrew and Bahn 1991). A datum point was selected and a grid system formed with respect to an N-S/E-W axis.

The grid system consisted of one meter plots. Alternating squares were chosen for sampling. A 50 centimeter2 test unit was excavated in each of the alternate plots. A large number of the plots were located in rough terrain consisting of large stones and extremely rocky soil. Small samples from each 50 centimeter test unit were screened through 1/8 inch mesh and collected at the site. Larger samples were bagged and returned to the laboratory for screening. Rocks were overturned at random in the selected one meter squares to search for puparia.

Excavations were also conducted under the cages. On December 1st, the final day of the experiment, the cage containing the burned specimen was moved and a one meter excavation unit was established where the cage had been located. Using standard

15 archaeological techniques (Renfrew and Bahn 1991), the sod layer and subsoil were removed, screened, and the insect associated materials collected for laboratory analysis.

Insects: Identification

Insects were identified in the laboratory using standard keys such as Hall (1947),

Cole (1969), Borror and White (1970), Greenberg (1973), Oldroyd and Smith (1973),

Anderson and Peck (1985), and Borror et al (1989). Adult insects were then pinned while larvae were kept in Pampel's solution.

16 Results: Chapter 3

The insect succession results for this experiment are summarized below by the stage in which they occurred. Each decay stage is divided. The commencement and termination of the remains stage was the most variable and was determined by the relative amount of flesh remaining upon the pig. Species diversity of insects utilizing each of the stages are shown in Figures 7-11 for Diptera and Figures 12-15 for

Coleoptera. Insects present during each stage for each pig are listed in Tables 1 and 2.

The level of burning obtained for BSS was consistent with a Crow-Glassman Scale

(1996) (CGS) Level #1 bum with some elements of a CGS #2 bum. A level #1 bum has the characteristics of epidermal blistering and singed hair, typical of a smoke-inhalation death. A level #2 bum displays a degrees of charring. The bum results from the present study are in slight contrast to Avila and Goff (1998) who obtained only a level #2 bum only. Once the bum was completed, the smell of roasted skin was readily apparent, unfortunately providing an attractant to large carnivores such as bears and scavenging birds.

Fresh Stage (day 1)

Odor associated with this stage was consistent with that of fecal matter and urine as the subjects bowels discharged upon death. Anthomyiidae, Otitidae, Phoridae,

Fannidae, and Cynomyopsis cadaverina (Robineau-Desvoidy) (Family Calliphoridae) were present and are typically attracted to fecal matter (Borror and White 1970). More than one inch of rain fell during day one. Flying insects in general were not visible during rainfall but increased markedly once the sun shone. Any eggs laid were likely washed away during the intermittent rains as none were present in the evening.

17 Adults of Phormia regina (Meigen) and a species of Sarcophaga were the most abundant decomposers sampled on the pigs during this stage, with adults of Eucalliphora

(Townsend) and Lucilia illustris (Meigen) contributing roughly an equal ratio. Flies of the species Protophormia terraenovae (Robineau-Desvoidy) were also present in low numbers at the burnt carcass while Anthomyiidae were present only at the control carcass. Additional Diptera present, albeit contributing very few adults to the whole community, were Phaenecia cadaverina, several species of Fannia, and members of the families Tachinidae, Heleomyzidae, and Phoridae. See Tables 1 and 2 for records of

Diptera present or absent on each respective carcass. The orders Hymenoptera and

Lepidoptera were observed flying around the pigs. Members of the families

Megachilidae and Vespidae were observed in the area but did not interact directly with the carcasses. The bulk of the fauna collected were present on both carcasses.

A slight difference in carcass color was apparent on the abdomen of the burnt carcass, which exhibited a large turquoise green di scoloration and lack of hair on the singed skin. In the evening, this discolored skin began to split and crack.

Bloated Stage (days 2,3-8)

The bloated stage commenced as gasses began to inflate the carcasses after day two. This stage began about a day earlier, and ended roughly a day sooner, for the burnt carcass possibly due to cracks in the skin assisting in carcass deflation. The axial portion of the burnt carcass became extremely dry and firm following the bum and displayed the least amount of insect activity. Unidentified eggs were present in the early days of this stage. Larvae of Ph. regina, the only immatures found during this stage, were present by

18 day five. Eggs and larvae were encountered in the mouth, ears, eyes and open body

cavity (at the end of the bloat).

Adults of Eucalliphora and Calliphora terraenovae were present in low numbers

while adults of Prot. terraenovae, two species of Sarcophaga and Fannia (burnt carcass)

were slightly more abundant. As bloating proceeded, Piophilidae adults of the species

Prochyliza brericomis (Melander) were attracted to each carcass. Sepsids, Sepsis spp.

and Meroplius Stercoronius (Robineau-Desvoidy), were also present on the burnt carcass.

Individuals of the genus Nemapoda were especially attracted to the control, albeit in low

numbers. A low number of Heleomyzids continued to show attraction to the control.

Coleoptera associated with the bloated stage were diverse. The burned skin on

the burned pig cracked and a small amount of fluid seeped from various cracks in the

abdomen and thorax. Members of the species Thanatophilus lapponicus (Herbst) were

active at these cracks in the skin and may have been feeding upon Diptera eggs. T.

lapponicus present at the control carcass were active on skin which began sloughing off toward the ground at the end of the bloat stage. Seven to eight days after death they were

seen mating on the burnt carcass.

High numbers of the genus Dermestes were found on the burnt carcass throughout the bloat period and were observed moving on the burnt carcass through the nose and mouth. Dermestes appeared to be attracted to fluid seepage from the nose and mouth of both carcasses. Dermestes were found in slightly lower numbers on the control.

Coleoptera present in moderate numbers included the families Histeridae and

Staphylinidae. Numbers of Chrysomelidae, Tenebrionidae, Elateridae (species 4), and

Scarabaeidae, subfamily Aphodiinae were low and nearly insignificant on the burnt

19 carcass. Of these, only Chrysomelidae and Scolytidae were present on the control.

Overall, more species of Coleoptera were associated with the burnt carcass. However,

Coleoptera were found in higher numbers on the control. The only Hymenoptera found on the control carcass were from the family Formicidae. Hymenoptera collected from the burnt carcass included Formicidae, Megachilidae (subfamily Megachilinae), and

Vespidae (subfamily Vespinae).

Red mites (order Acari) were first observed during this stage and were present on both carcasses for the remainder of the study. The mites may be phoretic on Histeridae as they were often observed on hister beetles as well as moving over the carcass.

Springett (1968) found that the mite Poecilochirus necrophori may have a symbiotic relationship with the beetle Necrophorus. Mites used Necrophorus adults as transport to a carcass and fed upon dipteran larvae alongside Necrophorus. By aiding in reduction of

Dipteran larvae, Necrophorus was more successful in breeding. In experiments where mites were absent, Necrophorus was entirely unsuccessful in reproducing. The mites were not observed to directly feed upon Dipterous larvae in the present study but may have a similar relationship with hister beetles.

Decay Stage (days 9-13)

Diptera associated with this stage were predominantly larval Ph. regina (Tables 1 and 2). Prot. terraenovae were present in very low numbers on both carcasses.

Sarcophaga (species 6) and Piophilidae (species 2) were found on the control carcass. In comparison, Fannia (species 1) was present only upon the burnt carcass.

Day 9 was marked by a very low temperature of 3°C. At this time adult Diptera activity was extremely low and the majority of the larvae present were in the body cavity

20 of the both pigs, generating enough heat to stay alive (Figure 2). Previous research has

shown that larval movement frequently raises internal temperatures above the ambient

temperature (Payne 1965, Williams and Richardson 1984, Early and Goff 1987, Catts and

Goff 1992). The low number of larvae on the burnt carcass enabled only a small number

to survive as the temperature approached freezing. The temperature rose on days 11 and

12 by 8°C and fresh eggs were present on both carcasses. Formicidae were observed

carrying off Diptera eggs while Staphylinidae preyed upon both Diptera eggs and larvae.

Histerids were observed preying upon both Dermestes and Diptera larvae underneath and

inside the orifices of the head.

As larvae consumed the organs and muscle tissue of each carcass, the skin

covering the abdomen became dry and provided a cover above the feeding larvae. Under

this cover on the burnt carcass on day 11, mating Nicrophorus and Necrodes

surinamensis (Fabricius) (family Coleoptera) were observed walking upon the frothy

maggot mass in the burnt carcass. At the end of the decay stage, larvae of T. lapponicus

and adults of Histeridae and Dermestidae continued to be fairly common on both

carcasses. N. surinamensis adults and larvae were only found on the burnt carcass at this

stage. Staphylinidae and Histeridae continued to prey upon larvae on both carcasses.

Hymenoptera present throughout the decay stage include Vespidae (subfamily Vespinae)

and Formicidae.

Post-Decay Stage (days 14-37)

The appearance of each carcass was modified by the feeding and migration of

larvae. The once fleshy abdomen became devoid of insects with the exception of

Coleoptera and various Hymenoptera. Most skin had fallen away from the thoracic

21 cavity and abdomen revealing the ribs and vertebrae. The skin of the burned carcass was stiff in areas but brittle in others. A maggot mass was observed in the abdomen of each carcass and exhibited a bubbly frothy mixture of larvae at the height of larval abundance and internal temperature readings. Small maggot masses were observed moving in the mouth, nose, ears, and eyes following the migration of the majority of larvae from the abdomen. Larvae consumed nearly all muscle tissue along the axial skeleton, shoulder joints, and pelvis of each carcass (Figures 3 & 4). Coleoptera often remained beneath each carcass and were only observed when the carcass was lifted.

At the commencement of this stage, Diptera larvae exited the carcasses after the majority of the flesh had been removed. Migrating larvae of Ph. regina and L. illustris exited each carcass beginning on day 13 for the control carcass and day 14 for the burnt carcass, often in full sunlight at midday. Eucalliphora adults were most abundant during this stage on the burnt carcass and were exceeded in numbers only by adults of Ph. regina. Larval migration was nearly complete by day 32 and finished by day 37 on each carcass. Genera present in moderate numbers on the burnt carcass were Eucalliphora and

Fannia (Species 1). Various Sepsidae were collected at each carcass and are listed in

Tables 1 and 2. One mating pair from the genus Nemapoda was captured on July 9th.

The Dipterans, Phaenecia sericata (Meigen), Phaenecia caeruleiviriais

(Macquart), Fannia canicularis (Linnaeus), F. difficilus (Stein), Fannia (species 3),

Piophilidae (species 1), made their first and only appearance during this stage at their respective carcasses as shown in Tables 1 and 2. Additional Diptera adults attendant for a brief time are all remaining families listed in Tables 1 and 2.

22 The most abundant adult species during this stage were the Piophilidae species

Proch. brericomis and Proch. xanthostoma (Walker). Mating pairs of Proch.

xanthostoma were captured from day 17 until day 27. Proch. brericomis mating pairs

were captured from day 29 until day 111, in the period directly following Proch.

xanthostoma.

Silphidae larvae of the species T. lapponicus and N. surinamensis increased in

abundance during the post-decay stage. Both species moved through the Diptera maggot

mass but appeared to be most interested in the dried skin of the carcasses. However, after

extended observation, both species were observed preying and fighting over dipteran

larvae. Adults of I lapponicus, Nicrophorus, Dermestidae, Histeridae, and

Chrysomelidae were observed on each carcass with only slight changes in population

numbers since the decay stage.

T. lapponicus were also observed preying upon Dipteran larvae. By day 28, a

copious amount of empty Coleopteran larval skins could be found in and surrounding both carcasses. Coleopteran families such as Staphylinidae, Coccinellidae, Curculionidae

(subfamily Brachyrhininae), Salpingidae, Scarabaeidae (subfamily Scarabaeinae),

and Elateridae (species 1 and 2), were present in diminished numbers on their respective

carcasses.

Of the numerous Hymenoptera collected, only the families Formicidae, Sphecidae

(subfamily Sphecini), Apidae (subfamily Apinae tribe: Bombini), and Vespidae

(subfamily Vespinae) displayed an active interest in the carcasses. Specidae and

Formicidae were numerous throughout this stage. Formicidae in particular w,ere

observed preying upon migrating larvae more often than upon feeding larvae. At the

23 height of larval migration, virtually every ant visible was carrying either Dipteran or

Coleopteran larvae.

Upon the completion of larval migration, the numbers of ants observed near the carcasses dwindled. Apidae (subfamily Apinae tribe: Bombini) exhibited aggressive behavior on various Coleoptera, namely Dermestidae and Histeridae. Araneida

(subfamily Araneae) also predated upon both adult and immature Diptera.

Remains (days 38+)

At the onset of this stage only bone, cartilage, dried skin, and a small amount of tissue was left on the carcasses (Figures 5 & 6). The rotten smell previously pervading the site became identifiable only on wet or unusually warm days.

Ph. regina continued to be the primary calliphorid found on the remains, albeit in small numbers. Prot. terraenovae was found on the burnt carcass in extremely low numbers and constitutes the only other calliphorid present. Sarcophagidae and Fannia were now more abundant than Calliphoridae. Four species of Sarcophagidae were found until the end of September with a few teneral adults still collected on day 60. Members of various Fannia spp. were also found in relatively moderate numbers throughout the end of September. Newly emerged flies were collected on day 56 on the burnt carcass.

F. canicularis was found sporadically throughout July and August on both carcasses.

Two species of Piophilidae also were active during this stage. Specimens of

Proch. xanthostoma were found mating in great numbers in mid-July, after which their numbers tapered off. In early October, their numbers increased a second time, and thereafter adults were not found again. Specimens of Proch. brericomis were found in moderate numbers from mid-July until early October. They were observed mating and

24 collected as late as day 105 (early October) on the burnt carcass. Piophilidae larvae were

collected from the control carcass between days 56-83. Unfortunately, they were not

identified to species and it is unclear whether they are Proch. xanthostoma or Proch.

brericomis. Each of these Piophilidae species were vital to the analysis of Diptera

activity during the remains stage as the adults were the principal species observed mating.

The latest seasonal observations of mating were of Heleomyzidae. Mating was

observed to occur from day 116 to day 138 (ie early October to early November) on both

carcasses. Immatures were not observed. Diptera present on both carcasses in low

numbers were Sepsidae ( Sepsis spp.) and Tachinidae spp (Tables 1 and 2).

Coleopteran larvae of T. lapponicus and N. surinamensis were essentially absent

during this stage. The final larval stages for these two species were collected in early

August at the same time Dermestes larvae became numerous. Larvae of T. lapponicus were present on each carcass for approximately 35 days. Anderson (1982) has

commented that the completion of larval stages of T. lapponicus lasts between 25-29 days. The departure of T. lapponicus in the beginning of August is not unexpected as

Anderson and Peck (1985) have remarked that this species decreases in numbers later in the year and overwinters.

By day 46,Dermestes larvae were abundant underneath the carcasses and inside the body cavity where the skin had dried. One such area consisted of the carcasses' dried

ears, which had been cleared of flesh, and some cartilage, leaving only dried skin husks.

Dermestes larvae were collected until day 67 for the control and day 70 for-the burnt

carcass. Thereafter, in reduced numbers, only adult Dermestes were found until day 97.

25 The greatest number of species were found in the remains stage for each carcass.

A possible reason for this diversity is the long duration of the stage, which can range from months to years, depending upon the environment. The rate at which the carcass decays is determined more by vertebrate scavengers, weather, etc. than by insects, and therefore the successional value of insects during this stage is limited. Adults of

Dermestes, T. lapponicus, Staphylinidae, and Histeridae were still present. Additional

Coleoptera associated with the remains stage for the control carcass but not for the burnt carcass are: Scarabaeidae (subfamily Troginae) and Curculionidae (subfamily

Brachyrhininae). The remaining Coleoptera found were present throughout the study and are listed in Tables 1 and 2.

Archaeology

All insect pupal evidence was found in the topsoil 10-30 centimeters below the surface. Pupal cases were collected from only two 50 cm test units. Three pupal casings were found farther than 3.3 meters from the carcasses. The farthest pupal casing was found 17 meters from the datum point. No puparia were found under rocks overturned at random in the selected one meter squares. In contrast, dozens of pupal casings were

.•j recovered from the one m excavation unit placed under the cage of the burned pig.

Vertebrate Carnivores and Scavengers

Damage to the carcasses was minimal on occasions when vertebrate carnivores and scavengers attempted access to the pigs due to the strength of the rebar cages. On day 2, bear droppings were found in the grass between the two cages and several pieces of rebar on the control cage were bent outwards. The pig inside was cut open slightly in the pelvic region, which enlarged a split in the skin already present due to the bloating

26 process. On day 6, more bear activity occurred on the west-facing side of the control

cage. Rebar was peeled back in several places but the integrity of the cage was intact and

the pig was not damaged.

Bear activity was absent throughout the decay and early post-decay stages but on

day 105, during the remains stage, damage occurred to both cages. The door of the cage

containing the control carcass was battered and the east-facing side of the burnt pig's cage

had rebar peeled back in one spot. Damage to the carcasses themselves was insignificant

and the study continued uninterrupted. The primary scavenger observed until the end of

the decay stage was the turkey vulture (Cathartes aura). Fortunately, the rebar was

welded in a close pattern and turkey vulture damage was kept to a minimum.

Larval Development

Dipteran Larvae

Ambient temperature averaged 22°C as a high and 7°C as the low on the day when larvae were first observed. Thereafter maximum temperatures averaged 20.3°C and minimum temperatures averaged 7°C until the first larval migration. The average maximum temperature during the thirteen day migration period was 26°C and the minimum low averaged 9°C. If the maggot mass is large enough and produces adequate heat for development only extreme variances in ambient temperature should negatively affect the larvae. The high ambient temperature during this two week post-peak internal temperature migration period averaged 18.5 °C with an average low ambient temperature of 6.5°C. These ambient temperatures were within a range that permitted an adequate environment for larval growth (Kamal 1958). Rainfall recorded during this period at the

27 site was 2.81 inches. Dew was present every morning and the majority of the daytime hours were partly cloudy.

When larvae first appeared on day 5 the average humidity was 86%. Days 6-13 had an average humidity of 74%. These are the eight days in which larvae developed through the first and second instar stages. Day 13 was the first time larvae were observed migrating from the control. Days 14-28 have an averge humidity of 72%. These are the fourteen days during which larvae concluded their migration. During times of humidity extremes, such as 96% and 20%, and temperature extremes, such as 3°C and 34°C, larvae appeared to become scattered and disorganized. The low ambient temperature had less of an effect at times when internal temperatures of the pigs were at a peak.

ControlS. scrofa

By day 5, approximately one dozen 3rd instar and two dozen 1st and 2nd instar Ph.

Regina larvae were collected. Numbers increased through days 9 and 10. On days 13 and

14 all three larval instars were found on the carcass but large masses of 3rd instar Ph. regina were predominant. On day 13, the first larval migration was observed, 12 days after the first eggs were laid. Larvae could be found exiting the carcass as late as day 24.

By day 32, newly eclosed adults were found.

Internal temperatures peaked at 30°C on day 13, the day of the first larval migration. Larvae continued to migrate as the internal temperature stabilized at an average of 25°C over the next seven days. On day 20, the internal temperature peaked again at 35°C conincident with a 3rd instar larval migration. A few scattered larvae continued to migrate until day 32. Larval migrations from the control pig correspond roughly with those from the burnt carcass. Migration of larvae from the control pig

28 began approximately a week before peak internal temperatures and continued for almost

two weeks. These differences are due to the higher internal peak temperature and greater

abundance of larvae in the burnt carcass

Burnt S. scrofa

By day 5, dozens of larvae were observed. Day 14 saw the first 3rd instar larval migration, 13 days after the first eggs were laid. Another larval migration occurred on

day 17. The number of larvae increased steadily until day 20, when large masses of 1st,

2nd, and 3rd instar larvae were found. On days 22-24, additional 3rd instar larvae were

found migrating. However, 3rd instar larvae were still found on, and leaving the carcass, on days 25 and 26. On day 28, there were virtually no dipteran larvae remaining on the carcass. By day 32, newly eclosed adults were observed, indicating a minimum immature development time of 31 days for Ph. regina in this environment. 3rd instar larvae of L. illustris are found on July 4th and were observed migrating between July 9th and 11th from both burnt and control carcasses. Larvae of Diptera other than L. illustris and Ph. regina were not detected.

Internal temperatures for the burnt specimen began at 7.8°C and peaked on day 11 at 26.7°C. The internal temperature peaked again on day 21 at 31.1°C. These peak internal readings only differed from the ambient temperature by 1-6°C. This peak temperature corresponds with the migration occurring on days 14 to 17 and on days 22 to

26, Each migration lasted approximately four days. Dillon (1997) has stated that calliphorid larval migration usually occurrs before the peak internal temperature has been reached. In this study, migration, began approximately a week before the highest peak internal temperature and continued for four days following the peak.

29 Coleopteran Larvae

Burnt and Control S. scrofa

Adults of the genus T. lapponicus were observed mating on days 6 and 8. Adults

of Nicrophorus and N. surinamensis were first observed mating on day 11. Coleopteran

larvae were also observed on this day but were not collected as the few present were being preyed upon by Histeridae and Formicidae. N. surinamensis larvae of all sizes

were found from days 11 -48 with a few larvae up to day 63. Small larvae of T.

lapponicus first appeared between days 7-26. Medium and large larvae of T. lapponicus were found between days 26-49. By day 28 in the burnt carcass and day 26 in the control

carcass hundreds of cast off Coleopterous larval skins were found inside, under, and

surrounding the carcasses.

Adults of the genus Dermestes were found from days 2-91. Immatures were found from days 46-70. Dermestes immatures were found two days before immatures of

N. surinamensis and T. lapponicus made their last appearance. This abrupt change from the high abundance of immatures of the latter two species to the high abundance of

Dermestes could be explained by the habits of each species. The immature developmental stages for Dermestes are not described here as individuals in this genus were not identified to species.

Temperatures under 10°C and over 35°C appeared to have the effect of lowering activity for coleopteran larvae as it did for dipteran larvae. In both carcasses, many coleopteran larvae were not only predaceous upon dipteran larvae, but upon newly eclosed flies as well.

30 Ratcliffe (1972) studied N. surinamensis in Nebraska. Adults were nocturnal.

Eggs were laid around large carcasses and hatched and began to feed 2-4 days later.

Anderson and Peck (1985) observed a total immature time of approximately 42-43 days.

These statistics correspond well with the data collected for the present study where the relatively few adults were collected in the early morning and evening and the maximum period of larval development was approximately 37 days.

The immature stage of T. lapponicus has been reported by Anderson (1982) as having two generations a year in Ontario, the first generation beginning in April-May and the second from early June-July. Complete development time was estimated at 32-33 days for each generation. The rate of development for T. lapponicus in the present study was approximately 23 days. Anderson (1982) estimated the egg stage to last 5-6 days.

Eggs were not identified, and therefore, were not included in the 23 day estimate. The immature stage for this insect in the present study may have lasted between 28-29 days.

The results found during this study are similar to the habits and natural history recorded for these T. lapponicus and N. surinamensis in the literature. Anderson and

Peck (1985) state that T. lapponicus is a cold-weather species that occurs at high mountain elevations in the west. The decrease in adults as the fall season progressed also concurs with the data of Anderson and Peck (1985) for the overwintering habits of this species.

The life cycle of flies is most commonly used to determine the Post Mortem

Interval (PMI) (Nuorteva 1977). However, with information on the immature stages of coleopteran species, it may be possible to use Coleoptera in conjunction with dipteran species for stronger PMI estimates.

31 Dillon (1997) found adults of both N. surinamensis and T. lapponicus. However,

no immatures were collected. This may be explained by 2 factors: (1) breeding

conditions may not have been optimal, and therefore, either breeding was not possible or

the immatures present were preyed upon or went unnoticed, and (2) these species could

have been outcompeted by D. talpinus, present in Dillon's study. In Dillon's sun habitat,

D. talpinus immatures and adults were found in both the decay and post-decay stages.

However, D. talpinus immatures were found only in the bloat stage for the shade habitat.

Therefore, competition between these Coleopteran species may only provide a partial

answer why differences in decomposition time was observed. The importance of season, temperature, humidity, and the amount of food resources available, may be significant factors in the ability of some immatures to survive to adults.

32 Discussion: Chapter 4

Kamal (1958) found Ph. regina have a pupal stage duration of 4 to 9 days and a

complete immature stage (egg to adult) of 10 to 12 days under laboratory conditions of

26+l°C and 50+2% relative humidity. Greenberg's (1991) study, conducted at 22°C,

found a pupal stage length of 5 days with approximately 13 days for total development

from egg to adult. The developmental averages in days presented by Kamal (1958) and

Greenberg (1991) are significantly less than those determined for this study. The temperature range provided by Greenberg (1991) is close to that of the present study.

The 50+2% relative humidity level used by Kamal (1958) is the optimal humidity found for Ph. regina larval development.

The early decay stages of fresh, bloat, decay, and post-decay are determined by the activity and rate of development of Diptera larvae (Early and Goff 1986). This provides an excellent opportunity for comparison of data in the present study and data collected by Dillon (1997). In the present study the burnt and control carcasses had nearly identical decay stage lengths, i.e., the onset of each decay stage varied only by approximately one day. Decay stage length was dependent upon factors such as insect predators and the lack of maggot mass cohesion on day 9 for the burnt carcass. The lack of cohesion may be related to the low ambient temperature and insufficient numbers of larvae present for creating optimal internal temperatures, thus, larval development was delayed. Not only did the carcasses in this study have nearly identical decay rates (+ 1 day) but they also mirrored the decay rates found by Dillon (1997).

When broken down numerically Dillon's (1997) table of decay stages maybe evaluated and approximately compared with the present studies numbers (Table 3).

33 While Dillon's experiment contrasted succession in pigs in sun and shade, the present

study contrasted burned and unbumed pigs in a partially shady site. However, these

differences are irrelevant because Dillon found no difference between the types of blow

fly species in shade vs. sun. Therefore, in order to compare her results with those of the present study the decay stage length in days for Dillon's sun/shade habitats were averaged in Table 3 with the results of the present study.

Some of the carcasses used in Dillon's study (1997) were clothed in various human apparel (shirts, etc.). Following the assumption that clothed carcasses retained

fluids longer and thus lengthened the stage in question (Dillon 1997). However, once the post-decay stage is reached, it is extremely difficult to determine as to when it concludes.

Therefore, this comparison of the length of the post-decay stage is only an approximation and rate of development of the insects involved should be used as the actual determinant of the length of a decay stage. With this in mind, the most abundant colonizing Diptera should be used to model insect succession.

The observed developmental rate of 31 days for Ph. regina does not take into account possible effects of competition which may have had a negative effect. A predatory environment combined with low nightly temperatures and the possibility of inadequate moisture all may have contributed to the lengthened larval developmental period. There was an abundance of fly egg predators present during the bloat and decay stages during which unidentified eggs were deposited. These included staphilinids, silphids, histerids, and Hymenoptera. Putman (1978) found that predation rates of immature insects of up to 66% may be reached in some instances. Nuorteva (1970) has also commented on the near extermination of Diptera eggs and larvae by histerid beetles.

34 Previous experiments have found that high numbers of predators and low average

temperatures can lengthen the decomposition progress (Johnson 1975, Nuorteva 1977,

Payne 1965 and Reed 1958).

Temperature and humidity were the principal factors affecting larval

development. Cloud cover also significantly lowered adult Diptera activity. The use of the interior Douglas-fir/ponderosa pine mixed habitat provided a good location to

contrast with that of Dillon (1997). While Dillon (1997) found eggs on day 1, in the present study the lack of Calliphoridae eggs on day 1 was probably the result of heavy rains. As in the present study, Dillon (1997) also found Diptera eggs in the fresh and bloated stages but not in the decay stage. Maggot masses were found between days 5-12

for both the burnt and control carcasses in the present study. This agrees with Dillon's results of 5-10 days for a similar interior Douglas-fir habitat.

The rate of development for Ph. regina was nearly identical on each carcass with the first migration of 3rd instar larvae occurring only a day apart. This pattern is followed closely byL. illustris. This is in contrast to findings by Dillon (1997), where Ph. regina and Prot. terraenovae were observed to be co-dominant on carrion where L. illustris was not. Ph. regina larvae of 1st, 2nd, and 3rd instars are found in the present study during the bloat, decay, and post-decay stages. In contrast Dillon (1997) found Calliphoridae 1st instars only in the bloat stage (sun) and fresh and bloat stages (shade), 2nd instars during bloat and decay stages (sun) and fresh and bloat stages (shade) and 3 rd instars during the bloat, decay, and post-decay stages in both sun and shade carcasses. Adul ts'of Ph. regina were present throughout both studies until the remains stage was reached, after which they were absent except in a few instances.

35 The presence of Ph. regina 1st and 2nd instars during later stages in the present

study indicates that eggs continued to be deposited well into the bloat stage and possibly

the decay stage. Each carcass, therefore, continued to provide a suitable substrate for the

deposition of eggs and their subsequent development into viable larvae. An important point noted by Kamal (1958) is that Ph. regina is not as susceptible to crowding, low humidity, etc. as the C. terraenovae (Macquart) and Phaenecia cadaverina that were also present. This adaptation may have aided Ph. regina to achieve its high level of

abundance over the latter two species.

Diptera

The succession of insects in the two pigs is summarized in Tables 1 and 2.

Species diversity was recorded in a logarithmic table for the burnt and control carcasses

separately (Figures 7-15). While some species were absent in one or the other carcass, they probably made little contribution to the decomposition process. Among the

Calliphoridae only Ph. regina made a significant contribution to this process. Ph. regina adults and immatures were most abundant among the order Diptera throughout the study until the post-decay stage when they shared high levels of abundance with L. illustris. It was also during the post-decay stage that Proch. brericomis and Proch. xanthostoma of the family Piophilidae and at least one species of Sepsidae were highly abundant.

Heleomyzidae were found mating in the remains stage.

The most abundant necrophagous species in this study was Ph. regina. The abundance of this blow fly at the site is not surprising considering its habits. It is a predominantly Rolarctic and rural species that has a seasonal appearance from the spring through the fall (Hall 1947, Greenberg 1973 and Anderson 1995) and is common at high

36 altitudes during these seasons. Kamal (1958) found that this blow fly was less sensitive

to crowding in a controlled environment than C. terraenovae and Cy. cadaverina. Each

of these species was present in small numbers in the present study, however, crowding may have influenced their relative abundance.

Cy. cadaverina is found from northern Labrador to the southern US border, however, it is most abundant along the Canadian-US border (Greenberg 1973). Although it has reproductive peaks in early spring and late fall, the conditions discussed above may have proved too negative, in terms of cold and predators, for this species to proliferate under the circumstances.

It was surprising to find such low numbers of C. terraenovae during the present study. This blow fly is most common in the northern United States and southern Canada, especially the Rocky Mountains. The seasonal activity pattern for this species is from late July to August (Hall 1947). The reason behind this blow fly's near absence is unexplained although temperature, overcrowding, and predation could be factors.

Temperature fluctuations and behavior relations for Ph. regina have been noted previously by Deonier (1940). Deonier found that this blow fly has a minimum activity temperature of 5°C. Several days in the first month of the study experienced temperatures near 5°C and Ph. regina larvae and adults were active in low numbers. This low activity level doubtlessly played a role in the extended developmental time observed for this blow fly.

L. illustris has been categorized by Hall (1947) and Greenberg (1973) as an open woodland and meadow blow fly species. Both Anderson (1995) and Greenberg (1973) agree that it has a Holarctic distribution, from southern Canada to northern Mexico and is

37 both an. urban and rural species. L. illustris are attracted primarily to fresh carcasses and

only rarely to dung (Greenberg 1973). Ironically, in the present study the only adults

collected were, in fact, found on the first day of the study during the fresh stage. No L.

illustris were found again until the first immatures were sampled 17 days later. Due to

the absence of immatures at this time of any other blow fly species.

Cole (1969) has reported Prot. terraenovae in the western US at high altitudes. It may perhaps be viewed as the counterpart of Ph. regina at the study site used in the present study. It is predominantly an early spring species that occurs in Utah from

March-June at altitudes o f4,000 to 6,000 feet. It has also been found in Colorado at

7,000+ feet from July to August (Hall 1947). This species therefore has a more northern distribution than Ph. regina, and so it is not surprising that it was in lower numbers.

Two species of Piophilidae, Proch. brericomis and. Proch. xanthostoma, were found mating in July and adults were found through early October on both carcasses'.

The presence of these Piophilidae species is not surprising. Cole (1969) reported specimens of Proch. brericomis from Flathead Lake, MT and Yellowstone Park from

July through August. Cole (1969) found that Proch. xanthostoma occurs from Colorado to California and north to Alaska. These species were most abundant during the post­ decay stage and may contribute more to PMI calculations during this stage than other

Diptera present.

The primary difference in the succession pattern during the fresh and bloat stages for Sepsidae on the burnt and control carcasses. Succession during the decay stage was similar for both carcasses while the post-decay stage brought about the greatest divergences in species diversity. Calliphoridae species such as Phae. caeruleiviriais,

38 Phae. sericata, Prot. terraenovae, and C. terraenovae each showed a preference for one of the carcasses (Tables 1 and 2). Species of the families Fannidae, Sarcophagidae and

Sepsidae became numerous on both carcasses.

Coleoptera

Coleoptera were not present during the fresh stage. During the bloated stage the

species Dermestes and T. lapponicus were the most abundant. The major differences between coleopteran in the two carcasses were the types and population sizes of predators of dipteran larvae. The only contrast between the insects on the two pigs during the decay stage was the absence of N. surinamensis and Nicrophorus on the control carcass.

Coleoptera of the post-decay stage were of the same composition for each carcass, with the major obserable variation being in numbers of Staphylinidae and Histeridae predators.

Hymenoptera and Other Arthropods

The only contrast in presence of Hymenoptera, non-coleopteran and dipteran arthropods between carcasses, was in species of Reduvidae (order Hemiptera) and the order Araneida. Both of these groups are predaceous upon insects and have been observed in previous forensic entomology studies (Payne et. al. 1968). The majority of the Hymenoptera collected in the present study were incidental species that did not interact directly with the carcass or the necrophagous species present. However, some

Hymenoptera collected were predacious upon insect larvae and eggs (Formicidae and

Vespidae wasps) or parasitic insects (Ichneumonidae and Pteromalidae).

39 A single species of Lepidoptera was observed in the site area. Although it did not

appear to interact with the carcasses themselves, Anderson et. al. (1996) has reported that

one Lepidopteran in the family Nymphalidae was collected while feeding on carrion.

Competition and Successional Pattern

Competition occurs between insects when utilizing a limited resource (Birch

1957). Competition among Diptera may have occurred in the pigs. A geometric

distribution pattern can indicate that one or two species are highly abundant, and

comprise roughly two thirds of the total numbers of individuals (Kuusela and Hanski

1982). This pattern was found in the present study and is often found among carrion fly

communities (Nuorteva 1970, Groth and Reissmiiller 1973, Beaver 1977, Hanski and

Kuusela 1977, Palmer 1980).

Examination of the species diversity for each pig indicates that competition may

have played a role in the outcome of the successional pattern. Kneidel (1984) predicted that large carrion should have high levels of Diptera competition with low species

diversity among decomposition stages. Diptera species diveristy was high during every

stage in the present study. A high species diversity in the presence of a large carcass is to be expected (Schoenly and Reid 1983). A possible explanation is that each carcass was large enough to provide adequate niches and resources for a higher number of species.

While the size of the carcass would seem to be the dominant factor in the discussion of species diversity, one must remember that the variation in environmental conditions, site diversity, and baseline populations of colonizing insects are also significant factors to consider.

40 Gilbert and Bass (1967) have reported finding pupae of Callitroga spp. and Ph.

regina in 130-160 year old Arikara burials. With the knowledge that both families of

blow fly appear by late March in the area of Gilbert and Bass's (1967) excavations and

are gone by mid-October the authors could determine when the burials were made.

Using this information, an archaeologist could determine the seasonal use of a site.

Burial practices may also be ascertained from this evidence. If a body was laid out for

defleshing or set out for a time until burial could take place, blow flies may have time to

lay eggs, complete a number of instars, or even reach the pupal stage.

Greenberg (1990) found Ph. regina postfeeding larvae pupate close to the carcass

and do not migrate farther than 3.3 m from the food site. In the present study, a puparium

was found 17 m from the datum point and it could be speculated that this pupal case was

from a different larval migration and was not associated with the study pigs. Dozens of pupal casings were found in the immediate vicinity of the carcasses. The majority were located underneath the carcasses in the topsoil. Unfortunately, the species to which the pupal casings belong is undetermined. But with the predominance of Ph. regina larvae, and the postfeeding behavior typical of the species, it is most likely that the majority are from this blow fly.

41 Conclusions-Chapter 5

The first hypothesis tested in this study was that the successional fauna in pig carcasses present in western Montana will be similar to that of pigs used by Dillon

(1997), who conducted a forensic entomological study in a similar biogeoclimatic zone, to that used in the present study, located in British Columbia, Canada. The present case study was conducted in a similar biogeoclimatic zone of interior Douglas-fir with a mixture of ponderosa pine.

There were two principal factors to compare between the two studies. The first comparison is the type of Diptera species present. Ph. regina and Prot. terraenovae were the two most abundant species in Dillon's study. The present study had Ph. regina

(Meigen) and L. illustris as the two most abundant Calliphoridae with Prot. terraenovae being less abundant. Ph. regina is a urban and rural cold-weather species, common to high altitudes from the spring into the fall seasons. P. terraenovae has habits similar to

Ph. regina but is more common to northern regions of North America and is considered

Ph. regina's northern counterpart by Hall (1947). Ph. regina 1st and 2nd instar larvae were also found into the post-decay stage in the present study, much later than in Dillon's study. L. illustris is also urban and rural, but prefers habitats which are more open.

Reasons why L. illustris was not as abundant on Dillon's test subjects is unknown but may be due to competition between the two species.

Additional Diptera species diversity similarities include the presence of Fanniidae almost exclusively during the post-decay and remains stages for both studies. The primary contrast in Diptera activity was the early arrival of Piophilidae larvae oh Dillon's test subjects between days 21-26. In contrast, Piophilidae larvae were not found on

42 carcasses in the present study until day 56. These arrival times are both earlier than previously reported 3-6 months for Southern Canada and Europe (Smith 1986).

Decay rates were similar for both studies with only a 2-3 day difference between

stages and similar lengths of time for cohesive maggot masses (2 days). Also, no reduction in insect numbers and diversity was observed in either study as the carcasses became desiccated in the remains stage. Furthermore, test subjects for both studies were only colonized by rural blow flies. A final interesting point is that species diversity was roughly the same for both studies although the pigs in the present study weighed approximately 82 kilograms and Dillon's pigs weighed about one-third this amount.

Coleoptera families were similar for both studies but species similarity could not be determined, except in the case of the family Silphidae, as other Coleoptera were not identified beyond genus for the present study. The Silphidae species N. surinamensis, T. lapponicus, and genus Nicrophorus were found in both studies. The only contrast was that Dillon found no immatures of these two species while immatures were prolific in the present study. This could be due to many factors including predation or adverse environmental conditions for reproduction.

These parallels in decay rates and species diversity for two studies conducted in an interior Douglas-fir and Douglas-fir/ponderosa pine Rocky Mountain biogeoclimatic zone may or may not be coincidence. Although it would be precarious to directly extrapolate data from one region to the other for forensic purposes, one could possibly confirm case study data found in one region by comparing it with the baseline data presented here. Direct extrapolationof data may lead to errors until more information is compiled for comparison. This tentative confirmation with data collected in a similar

43 biogeoclimatic zone, yet hundreds of miles away, is a positive step for Montana forensic

entomological studies.

The second hypothesis states that the burned and unbumed specimens will have

different successional fauna specific to their physical states. The successional pattern

was very similar between the burnt and unbumt pigs. The rate of decay differed by

approximately one day during the decay stage and the co-dominant species present was the same for each pig throughout the study. The similarities between the conclusions of this bum study and the previous bum study by Avila and Goff (1998) are also not

surprising. Unfortunately, species types, successional patterns, and decay rates could not be compared due to an extreme contrast in biogeoclimatic zones (Hawaii and Montana) and indigenous species. From Avila and Goff (1998) and the present study it maybe concluded that carrion Diptera and Coleoptera are attracted to burned carrion as quickly as to non-bumed carrion. Future studies may determine if differences in accelerant use create differences in successional patterns of species diversity.

The third hypothesis for this study states that the insect successional pattern and species diversity on carcasses present in a mixed urban/rural and those present in a rural area will be distinct (Temenyl997). A comparison was made with results from the present study and those of Temeny (1997) who conducted a spring study in a mixed urban/rural environment. The decomposition stages of the Temeny study compared with those of the present study are presented in Table 4.

Temeny (1997) defines the end of the decay stage as when the greater part of the flesh has been removed. In Temeny's study maggots were visible by day 11 compared with day 5 in the present study. This developmental delay as compared with

44 developmental time in the present study may be due to cooler temperatures in the spring,

although no climatological data is presented in Temeny's study, making comparisons

difficult.

No specific statements can be made concerning a comparison of species diversity for

spring/summer habitats and urban/rural locations using Temeny's Montana study.

Because insects were not identified beyond the family level. All of the families collected in Temeny's study were also found in the present study but nothing more may be generalized with the data provided.

The present case study has provided new information to be included in the forensic entomology database for the state of Montana. The study has revealed numerous similarities in insect succession with an experiment performed in a similar biogeoclimatic zone located in British Columbia, Canada. While this information should not be directly extrapolated to forensic cases in Montana it does provide investigators with an additional database for use in forensic entomology cases in a similar environment. The results of the present bum study parallel those found by Avila and Goff (1998) and strengthen their decision that although a slight time difference occurs among the early decompositional stages, the fauna for both burned and unbumed carcasses remains similar. Case studies from both urban and rural forensic cases obtained from the Montana Forensic Science

Division have been, and will continue to be, analyzed for an ameliorated view of forensic entomology in the state of Montana.

45 Table 1 Insect Succession for the burnt Sus scrofa:

Stage and time Insect since death Order Familv Genus and species Stage

Fresh (day 1) Diptera Calliphoridae Phormia regina ad Eucalliphora ad Protophormia terraenovaead Lucilia illustris ad Sarcophagidae Sarcophaga sp. 1 ad Sarcophaga sp. 2 ad Sarcophaga sp. 3 ad Sarcophaga sp. 4 ad Fannidae Fannia sp. 1 ad Tachinidae sp. 1 ad sp. 4 ad Phoridae sp. uniden. ad Hymenoptera Megachilidae not collected Vespidae not collected Lepidoptera spe. 1 ad Bloat (days 2-8) Diptera Calliphoridae Phormia regiria ad, im Eucalliphora ad Protophormia terraenovaead Calliphora terraenovae ad Sarcophagidae Sarcophaga 1 sp. ad Sarcophaga sp. 2 ad Fanniidae Fannia sp. 4 ad Piophilidae Prochyliza brericomis ad Sepsidae Meroplius Stercoronius ad Sepsis sp. ad Coleoptera Silphidae Silpha sp. ad Dermestidae Dermestes sp. ad Scarabaeidae sp. uniden. ad sub: Aphodiinae Histeridae sp. uniden. ad Staphylinidae sp. uniden. ad Chrysomelidae sp. uniden. ad Tenebrionidae sp. uniden. ad Elateridae sp. 4 ad Hymenoptera Formicidae sp. uniden. ad Megachilidae sp. uniden. ad sub: Megachilinae Vespidae sp. uniden. ad sub: Vespinae Acari sp. uniden.

46 Decay (days 9-13) Diptera Calliphoridae Phormia regina ad, im Protophormia terraenovaead Sarcophagidae Sarcophaga sp. 6 ad Fanniidae Fannia sp. 1 ad Piophilidae sp. 2 ad Coleoptera Silphidae Nicrophorus ad Necrodes surinamensis ad Thanatophilus lapponicusad, im Dermestidae Dermestes ad, im Histeridae sp. uniden. ad Staphylinidae sp. uniden. ad Hymenoptera Formicidae sp. uniden. ad Vespidae sp. uniden. ad sub: Vespinae Acari sp. uniden.

Post-decay (14-37) Diptera Calliphoridae Phormia regina ad, im (1st, 2nd, and 3rd instars) Eucalliphora ad Protophormia terraenovaead Lucilia illustris ad, im (3rd instar) Calliphora terraenovae ad Phaenecia caeruleiviriais ad Sarcophagidae Sarcophaga spe. 1 ad Sarcophaga spe. 5 ad Fanniidae Fannia canicularis ad Fannia difficilus ad Fannia sp. 1 ad Piophilidae Prochyliza brericomis ad Prochyliza xanthostoma ad sp. 1 ad sp. 2 ad Sepsidae Meroplius Stercoronius ad Sepsidomorph sp. uniden. ad Tachinidae Dexinae sp. uniden. ad sp. 1 ad Anthomyiidae sp. uniden. ad Chloropidae sp. uniden. ad Empididae sp. uniden. ad Rhagionidae sp. uniden. ad Coleoptera Silphidae Nicrophorous sp. uniden. ad Silpha sp. uniden. ad Thanatophilus lapponicusad,im Necrodes surinamensis ad,im Dermestidae Dermestes sp. uniden. ad Scarabaeidae sp. uniden. ad sub: Scarabaeinae Histeridae sp. uniden. ad

47 Staphylinidae sp. uniden. ad Chrysomelidae sp. uniden. ad Coccinellidae sp. uniden. ad Elateridae sp. 1 ad sp. 2 ad Hymenoptera Formicidae sp. uniden. ad Ichneumonidae sp. uniden. ad Colletidae sp. uniden. ad sub: Hylaeinae Sphecidae sp. uniden. ad sub: Sphecini Megachilidae sp. uniden. ad sub: Megachilinae Apidae sp. uniden. ad sub: Apinae tribe: Bombini Vespidae sp. uniden. ad sub: Vespinae Araneida sub: Araneae sp. 1 ad Acari sp. uniden. ad

Remains (days 38+) Diptera Calliphoridae Phormia regina ad Protophormia terraenovaead Sarcophagidae Sarcophaga sp. 1 ad, im Sarcophaga sp. 3 ad Sarcophaga sp. 4 ad Sarcophaga sp. 5 ad Fanniidae Fannia canicularis ad Fannia sp. 1 ad Fannia sp. 2 ad Fannia sp. 5 ad Piophilidae Prochyliza brericomis ad, im Prochyliza xanthostoma ad, im Sepsidae Sepsis sp. uniden. ad Tachinidae Dexinae sp. uniden. ad sp. 1 ad sp. 2 ad sp. 5 ad Drosophilidae sp. uniden. ad Trixoscelididae sp. uniden. ad Otitidae sp. uniden. ad Heleomyzidae sp. uniden., mating ad Anthomyiidae sp. uniden. ad Empididae sp. uniden. ad Coleoptera Silphidae Silphus sp. uniden. ad Thanatophilus lapponicusim Necrodes surinamensis im Dermestidae Dermestes sp. uniden. ad, im Staphylinidae sp. uniden. ad Histeridae sp. uniden. ad Chrysomelidae sp. uniden. ad

48 Elateridae sp. 3 ad Nitidulidae sp. uniden. ad Hymenoptera Formicidae sp. uniden. ad Ichneumonidae sp. uniden. ad Sphecidae sp. uniden. ad sub: Sphecini Apidae sp. uniden. ad sub: Apinae tribe: Bombini Vespidae sp. uniden. ad sub: Vespinae Vespidae sp. uniden. ad sub: Eumeninae Andrendiae sp. uniden. ad sub: Panurginae Andrenidae sp. uniden. ad sub: Andreninae Superfamily Chalcidoidea Pteromalidae sp. uniden. ad Hemiptera Reduvidae sp. 1 ad sp.2 ad sp.3 ad sp.5 ad Araneida Araneae sp. 1 ad sp.5 ad Acari sp. uniden. ad

49 Table 2 Insect Succession for the control Sus scrofa:

Stage and time Insect since death Order______Family______Genus and species______Stage

Fresh (day 1) Diptera Calliphoridae Phormia regina ad Eucalliphora ad Protophormia terraenovaead Lucilia illustris ad Phaenecia cadaverina ad Sarcophagidae Sarcophaga sp. 1 ad Sarcophaga sp. 2 ad Sarcophaga sp. 3 ad Sarcophaga sp. 4 ad Faniidae Fannia sp. 2 ad : Heleomyzidae sp. uniden. ad Anthomyiidae sp. uniden. ad Phoridae sp. uniden. ad Hymenoptera Megachilidae not collected Vespidae not collected

Bloat (days 2-8) Diptera Calliphoridae Phormia regina ad, im Eucalliphora ad Protophormia terraenovaead Calliphora terraenovae ad Sarcophagidae Sarcophaga sp. 1 ad Sarcophaga sp. 2 ad Piophilidae Prochyliza brericomis ad Sepsidae Nemapoda sp. uniden. ad Heleomyzidae sp. uniden. ad Coleoptera Silphidae Silpha spe. uniden. ad Dermestidae Dermestes sp. uniden. ad Histeridae sp. uniden. ad Staphylinidae sp. uniden. ad Chrysomelidae sp. uniden. ad Scolytidae sp. uniden. ad Hymenoptera Formicidae sp. uniden. ad Acari sp. uniden.

Decay (days 9-13) Diptera Calliphoridae Phormia regina ad, im Protophormia terraenovaead Sarcophagidae Sarcophaga sp. 6 ad Piophilidae sp. 2 ad Coleoptera Silphidae Thanatophilus lapponicusad, im Dermestidae Dermestes ad, im Histeridae sp. uniden. ad Staphylinidae sp. uniden. ad Hymenoptera Formicidae sp. uniden. ad

50 Vespidae sp. uniden. ad sub: Vespinae Acari sp. uniden.

Post-decay (14-37) Diptera Calliphoridae Phormia regina ad, im (1st, 2 , and 3rd instars) Eucalliphora ad Lucilia illustris ad, im (3rd instar) Phaenecia sericata ad Sarcophagidae Sarcophaga sp. 1 ad Sarcophaga sp. 5 ad Fanniidae Fannia canicularis ad Fannia sp. 1 ad Fannia sp. 3 ad Piophilidae Prochyliza brericomis ad Prochyliza xanthostoma ad Sepsidae Meroplius Stercoronius ad Nemapoda sp. uniden. ad (mating) Sepsis sp. uniden. ad Tachinidae sp. 1 ad Syrphidae sp. uniden. ad Lauxaniidae sp. uniden. ad Dryomizidae sp. uniden. ad Tabanidae sp. uniden. ad sub: Tabanus Diapriidae sp. uniden. ad Coleoptera Silphidae Nicrophorous ad sp. uniden. ad Silpha sp. uniden. ad Thanatophilus lapponicus ad,im Necrodes surinamensis ad,im Dermestidae Dermestes sp. uniden. ad Scarabaeidae sp. uniden. ad sub: Scarabaeinae Histeridae sp. uniden. ad Staphylinidae sp. uniden. ad Chrysomelidae sp. uniden. ad Curculionidae sp. uniden. ad sub: Brachyrhininae Salpingidae sp. uniden. ad Elateridae sp. 1 ad sp. 2 ad

Hymenoptera Formicidae sp. uniden. ad Ichneumonidae sp. uniden. ad Colletidae sp. uniden. ad sub: Hylaeinae Sphecidae sp. uniden. ad

51 sub: Sphecini Megachilidae sp. uniden. ad sub: Megachilinae Apidae sp. uniden. ad sub: Apinae tribe: Bombini Apidae sp. uniden. ad sub: Apinae tribe: Apini Vespidae sp. uniden. ad sub: Vespinae Halictidae sp. uniden. ad sub:Halictinae Andrendiae sp. uniden. ad sub: Panurginae Lepidoptera sp. 2 ad Acari sp. uniden.

Remains (days 38+) Diptera Calliphoridae Phormia regina ad Sarcophagidae Sarcophaga sp. 1 ad, im Sarcophaga sp. 3 ad Sarcophaga sp. 4 ad Sarcophaga sp. 5 ad Fanniidae Fannia sp.1 ad Fannia sp. 2 ad Piophilidae Prochyliza brericomis ad, im Prochyliza xanthostoma ad, im Sepsidae Sepsis sp. uniden. ad Tachinidae Dexinae sp. uniden. ad sp. 1 ad sp. 2 ad sp. 3 ad sp. 4 ad Trixoscelididae sp. uniden. ad Otitidae sp. uniden. ad Heleomyzidae sp. uniden., mating ad Syrphidae sp. uniden. ad Coleoptera Silphidae Silphus sp. uniden. ad Thanatophilus lapponicus im Necrodes surinamensis im Dermestidae Dermestes spec, uniden. ad, im Staphylinidae sp. uniden. ad Histeridae sp. uniden. ad Chrysomelidae sp. uniden. ad Lygaeidae sp. uniden. ad Elateridae sp. 3 ad Nitidulidae sp. uniden. ad Scarabaeidae sp. uniden. ad sub: Troginae Curculionidae sp. uniden. ad sub: Brachyrhininae Hymenoptera Formicidae sp. uniden. ad

52 Ichneumonidae sp. uniden. ad Colletidae sp. uniden. ad sub: Hylaeinae Sphecidae sp. uniden. ad sub: Sphecini Megachilidae sp. uniden. ad sub: Megachilinae Halictidae sp. uniden. ad sub:Halictinae Andrendiae sp. uniden. ad sub: Panurginae Andrenidae sp. uniden. ad sub: Andreninae Super Chalcidoidea Pteromalidae sp. uniden. ad Hemiptera Reduvidae sp. 1 ad sp. 2 ad sp. 4 ad Araneida Araneae sp. 1 ad sp. 2 ad sp. 3 ad sp. 4 ad sp. 6 ad Acari sp. uniden. ad

53 Table 3

Comparison of decay stage lengths from Dillon (1997) and the present study:

Decay Stage______Length in days: Dillon . Length in days: Present Study

Fresh 2 1 Bloat 5 7 Decay 8.5 5 Post-Decay 65 77

Table 4

Comparison of decay stage lengths from Temeny (1997) and the present study:

Decay Stage______Length in days: Dillon ______Length in days: Present Study

Fresh 2 1 Bloat 7 7 Decay 65 5 Post-Decay 25 77

54 Figure 1

Species Diversity, FFesh: Burnt 1.000 I. Phonria regina 2.SarcaSp.l 3. Protaphorrria terraenovae 4. Ludlaillustris 5. Eucalliphora 6. Saim Sp. 2 7. Saim Sp. 3 8. Saim Sp 4 9. Farniia Sp 1 10. Tachinidae Sp. 1 II. Tachinidae Sp. 4 0.001 9 10 11 Types of Spedes

Species Diversity, Fresh: Control

* * 1.000 L Phormia regina 2 Sarca Sp 1 4. Lutilia Oiusrtris 5. Eucalliphora 6 S a rc o S p 2 7. S arco S p 3 &SarcaSp. 4 12Phaeneda cadaverina 13. Fannia S p 2 14 Hdeopmyzidae 15. Anthomyiidae 16 Phoridae 0.001 1 2 4 5 6 7 8 12 13 14 IS 16 Types o f Spedes

55 Figure 2

Species Diversity, Bloated: Burnt

1. Phormia regina 2.SarcaSp.l 3. Protophormia 5. Eucalliphora ft 0.100, 6. Sarca Sp. 2 17. Prochyliza brericomis 18. Chlliphora terraenovae 0.010 : 19. Fannia Sp. 4 20. Memphis stercomis 21. Sepsis Sp.

1 2 3 5 6 17 18 19 20 21 Types of Spedes

Species Diversity, Bloated: Control 1.000 1. P. regina 2. Sarca Sp. 1 3. Protophormia 5. Eucalliphora g 0.100 14. Heleomyzidae 17. Prochyliza brericomis 18. Calliphora terraenovae 22. NemannHn

3 5 14 17 18 22 Types of Species

56 Figure 3

Species Diversity, Decay: Burnt

1.000 ? 1. Phormia regina

Protophormia terraenovae

0.010 :

0.001

Types of Species

Species Diversity, Decay: Control

1 .0 0 0 q 1. P. regina

Protophormia terraenovae S 0.100 : 23. Sarca Sp.

24. Piophilidae

0.0 1 0 :

0.001

Types of Species

57 Figure 4

1. Phormia regina 2 .S a r ca S p l 3. Protophonria Spedes Diversity, Post-Decay: Burnt 4. LucillaOlustris 1.000 5.Eucafliphna 9. Fannia Sp 1 sa 10. Tachinidae Sp 1 O4) 15. Anthontyiidae aU 17. Prochyliza breritaniis a. 0.100 18. Calliphora terraenovae uV a 20. Merioplius stericomis rt 25. Phaeneciacaeruldviriais -Osa a 26. Sarax Sp5 A < 27. Fannia canktdaris

Species Diversity, Post-Decay: Control 1. P. regina 2. Saim Sp 1 5JEucalliph. 9. Fannia S p l « 0.100 : 10. Tachinidae Sp 1 17. Prochyliza brericomis 20. Mencius < 0.010 : stercoroniiK 22.Nemapoda 29. Prochyliza xanthostoma 0.001 37. Syrphidae 38. Lauxaniidae 1 2 5 9 10 17 20 22 29 37 38 39 40 41 39. Diyorayz. Ttypes of Spedes 40. Tabanidae

58 Figure 5

I. Fhomia regina Spedes Diversity, Remains: Burnt 2. Sarcophaga S p l 3. ftotophorrria 10. Tachinidae sp. 1 II. Tachinidae Sp 2 14.Hdeomradae 15. Anthonwiidae 17. Prochyliza brericomis 21. Sepsis 27. Fannia canicularis 29. Prochyliza xairthostoma 31.Dexinae 42.Drosophilidae 43. Irixoscdididae <' 44. Otitidae 1 2 3 10 11 14 15 17 21 27 29 31 42 45 44 45 46 47 45. FanniaSp. 5 Types of l^redes 46. Tachinidae Sp 5 47. Empididae

Species Diversity, Remains: Control

1.000 1. Phormia regina 2. Sarcophaga Sp. 1 a 7. Sarcophaga Sp. 3 8 8. Sarcophaga Sp. 4 8 9. Fannia Sp. 1 An a> 0.100 10. Tachinidae S p .l « 11. Tachinidae Sp. 2 «a 13. Fannia Sp. 2 T9a 14. Heleomyzidae s 15. Tachinidae Sp. 3 .o 17. Prochyliza < 0.010 brericornis 21. Sepsis 26. Sarcophaga Sp. 5 I 29. Prochyliza £ xanthostoma 31. Dexinae 0.001 37. Syrphidae 8 9 10 11 13 14 15 17 21 26 29 31 37 43 43. Trixoscelididae Types of Species

59 Figure 6

Spedes Diversity, Bloated: Bunt

1. Dermestes 2 Thanatophilus lapponiciB 3. Staphyliridae 4 Scarabaeidae sub: Aphodiinae 5. Lfistseridae & Onysomelidae 7. Tenebrionidae & Elateridae Sp. 4

1 2 3 4 5 6 7 8 Types of Species

Species Diversity, Bloated: Control 1.000

£ 01100:

Types of Species

60 Relative Abundance Percent Relative Abundance Percent o

M 7 Figure

Sn i» H r Figure 8

Spedes Diversity, Post-Decay: Burnt

I. Denrcstes Z ThanatopKliB bpporiciE 3. Stapfyiimbe 5. Ifcteridae 6 , QrvwrreBdae II. NcropinroiB 1Z Scarabaeidae sife Scarabaeirae 13. Cbcrinellidae 14. Qeterkbe Sp 1

1 2 3 5 6 11 12 13 14 Types of Spedes

Spedes Diversity, Post-Decay: Control

I. Dernestes Z Thanatoplihs lagporiaB 3 . Staphyliiiclae 5. Ifcteridae 6. Chrysorrelidde II. NcrophoroiE 1Z Scarabaeidae 15.CUrctfioiidae 1& Saifingidae 17. Elateridae Sp 2

1 2 3 5 6 11 12 15 16 17 Types of Spedes

62 Figure 9

Species Diversity, Remains: Burnt

1. Denmestes ZlhaiatqMiK la(pricus 3. Stafiryiini(fae 5.Hsteridae 6.Girysameli(be 18.HatericbeSp3 19. NMdiicfae 20.0titidae

1 2 3 5 6 18 19 20 'types of Spedes

Spedes Diversity, Remains: Control

1.000 1. Dermestes 2. ThanatophiliR Lapponicus 3. Staphylinidae S. Ifcteridae (■.Chysomelidae 15. CknuKoradae 18. Elateridae Sp 3 19. NitxMidae 2L Scarabaeidae < 0.010 adx Ttoginae 22.Ljgaeidae

0.001 2 3 5 6 15 18 8 21 22 Types of Spedes

63 Temperature in Celcius Temperature in Celcius 0 i 50 \ * S •—AmbetTmp o — AmbetTmp Hih nenlTmp. Tem Internal - A - igh H p. Tem bient m —A —♦ Low p. Tem bient — m A —• — • —A m bient Tem p. Low —♦—A m bient Tem p. H igh —A—Internal Temp. —A—Internal igh H p. Tem bient m —♦—A Low p. Tem bient m —A —• 4 8 0 2 4 6 8 0 2 4 6 8 30 28 26 24 22 20 18 16 14 12 10 8 6 4 2 4 8 0 2 4 6 8 0 2 4 6 8 30 28 26 24 22 20 18 16 14 12 10 8 6 4 2 VA rV l ime Since Death in Days in Death Since ime l Time Since Death in Days in Death Since Time Temperature: CSS Temperature: Temperature: BSS Temperature: Figure 10 Figure 64

- 0 2 H- References

Adair, T.W. Three species of blowfly (Diptera: Calliphoridae) collected from a human stillborn infant in the Rocky Mountains of Colorado. Journal o f Medical Entomology 1999; 36(3):236-237.

Anderson, G.S. The use of insects in death investigations: An analysis of cases in British Columbia over a five year period. Canadian Society o f Forensic Sciences Journal 1995; 28:277-292.

Anderson, G.S. and S.L. Van Laerhoven. Initial studies on insect succession on carrion in southwestern British Columbia. Journal o f Forensic Sciences 1996; 41(4):617-625.

Anderson, Robert S. Resource partitioning in the carrion beetle (Coleoptera: Silphidae) fauna of southern Ontario: Ecological and evolutionary considerations. Canadian Journal of Zoology 1982; 60:1314-1325.

Anderson, Robert S. and Stewart B. Peck. The Insects and Arachnids o f Canada Vol. 13: The Carrion Beetles o f Canada and Alaska. Ottawa, Ontario: Biosystematics Research Institute. Research Branch, Agriculture Canada, 1985.

Avila, Frank W., and M. Lee Goff. Arthropod succession patterns onto burnt carrion in two contrasting habitats in the Hawaiian Islands. Journal o f Forensic Sciences 1998; 43(3):581-586.

Baumgartner, Donald L. The hairy maggot blow fly Chrysomya Rufifacies (MacQuart) confirmed in Arizona. Journal o f Entomological Science 1986; 21(2): 130-132. Spring Season Survey of the Urban Blowflies (Diptera: Calliphoridae) of Chicago, Illinois. The Great Lakes Entomologist 1988; 21:119-121. Abstract.

Beaver, R.A. Non-equilibrium "island" communities: Diptera breeding in dead snails. Journal of Animal Ecology 1977; 46:783-798.

Benecke, Mark. Use of forensic entomology in cases concerning putrefied corpses. Archiv fuer Kriminologie 1996; 198(3-4):99-109 (In German).

Bergeret, M. Infanticide, momification du cadavre. Decouverte du cadavre d'un enfant nouveau-dans une cheminee ou il s'etat momifie. Determination de l'epoque de la naissance par la presence de nymphes et de larves d'insectes dans le cadavre et par l'etude de leurs metamorphoses. Ann. Hyg. Publique Med. 1855;Leg. 4:442-452.

Birch, L.C. The meanings of competition. The American Naturalist 1957; XCI(856):5- 18.

65 Blackith, R.E. and R.M. Blackith. Insect infestations of small corpses. Journal o f Natural History 1990; 24:699-709.

Bomemissza, G. F. An analysis of arthropod succession in carrion and the effect of its decomposition on the soil fauna. Australian Journal o f Zoology 1957; 5:1-12.

Borror, Donald J., Charles A. Triplehom, and Norman F. Johnson. An introduction to the study o f insects. 6th edition, Harcourt Brace College Publishers, New York, 1989.

Borror, Donald J. and Richard E. White. A field guide to the insects o f America north o f Mexico. Houghton Mifflin Company, Boston, 1970.

Braack, L.E.O. Arthropods associated with carcasses in the Northern Kruger National Park. South African Journal o f Wildlife Research 1986; 16:91-98.

Brouardel, P. Determination de l'epoque de la naissance et de la morte d'un nouveau-ne a l'aide de la presence d'acares. Ann. Hyg. Med. Leg. 1879; 28:153.

Burger, J.F. Studies on the succession of saprophagous Diptera on mammal carcasses in southern Arizona. Masters's Thesis; University of Arizona, Tusco, AZ, 1965.

Catts, E. Paul. Analyzing entomological data. In Entomology and Death: a Procedural Guide. Joyce's Print Shop, Clemson, SC, 1990; 24-35.

Catts, E. Paul and Neal H. Haskell. Entomology and Death: A Procedural South Guide. Carolina: Joyce's Print Shop, Inc., 1990.

Catts, E. Paul and M.L. Goff. Forensic Entomology in Criminal Investigations. Annual Review o f Entomology 1992; 37:253-272.

Chu, H.F. and Wang, L.Y. Insect carcasses unearthed from the Chinese antique tombs. Aca Entomol. Sinica 1975; 18:333 (Chinese).

Coe, M. The decomposition of elephant carcasses in the Tsavo (East) National Park, Kenya. Journal o f Arid Environmental Research 1978;1:71-86.

Cole, Frank R. The flies o f western North America. University of California Press, Berkeley, 1969.

Comaby, B.W. Carrion reduction by animals in contrasting tropical habitats. Biotropica 1974; 6:51-63. de Stefani, T. The economic significance of applied entomology. Forensic Entomol. Allavamenti (Pallermo) 1921; 2:131. (Italian)

/

66 Deoneir, C.C. Carcass temperatures and their relation to winter blowfly activity in the Southwest. Journal o f Economic Entomology 1940; 33:166-170.

Dicke, R J. and J.P. Eastwood. The seasonal incidence of blowflies at Madison, Wisconsin (Diptera—Calliphoridae). Transactions o f the Wisconsin Academy o f Science, Arts & Letters 1952; 41:207-217.

Dillon, Leigh Celeste. Insect Succession on Carrion in Three Biogeoclimatic Zones o f British Columbia. Master's Thesis, Department of Biological Sciences, Simon Fraser University, B.C., 1997.

Early, M. and M.L. Goff. Arthropod succession patterns in exposed carrion on the island of O'ahu, Hawaiian Islands, USA. Journal o f Medical Entomology 1987; 23:520-531.

Erzin9lioglu, Y.Z. Few flies on forensic entomologists. New Scientist May: 1985:17.

Eyre, F.H. Forest cover types o f the United States and Canada. Society of American Foresters, Bethesda, MD, USA, 1980.

Fuller, M.E. Insect inhabitation in carrion: a study in animal ecology. Aust. Council Sci. Ind. Res. Bull. 1934; 82:1.

Galloway, Alison, Walter H. Birkby, Allen M. Jones, Thomas E. Henry, and Bruce O. Parks. Decay rates of human remains in an arid environment. Journal o f Forensic Sciences 1989; 34(3):607-616.

Gilbert, B.M. and W.M. Bass. Seasonal dating of burials from the presence of fly pupae. American Antiquity 1967; 32:534-535.

Glassman, David M. and Rodney M. Crow. Standardization Model for Describing the Extent of Bum Injury to Human Remains. Journal o f Forensic Sciences 1996; 41(1):152-154.

Goff, M.L. Unpublished case data; University of Hawaii at Manoa, Honalulu, HI, 1992a. Problems in estimation of postmortem interval resulting from wrapping of the corpse: a case study from Hawaii. Journal o f Agricultural Entomology 1992b; 9(4):237- 243. Estimation of postmortem interval using arthropod development and successional patterns. Forensic Science Review 1993; 5(2):82-94.

Goff, M.L. and C.B. Odom Early M. Estimation of postmortem interval by entomological techniques: a case study from Oahu, Hawaii, Bulletin for the Society o f Vector Ecology 1986; 11:242-246. Forensic entomology in the Hawaiian Islands, U.S.A. Three case studies. American Journal o f Forensic and Medical Pathology 1987; 8:45-50.

67 Goff, M.L., S. Charbonneau, and W. Sullivan. Presence of fecal matter in diapers as potential source of error in estimations of postmortem intervals using arthropod development patterns. Journal o f Forensic Science 1991; 36:1603-1606.

Goff, M. L., M. Early, C.B. Odom, and K. Tullis. A preliminary checklist of arthropods associated with exposed carrion in the Hawaiian Islands. Proceedings o f the Hawaii Entomological Society 1986; 26:53-57.

Goff, M.L., A.I. Omori, and J.R. Goodbrod. Effect of cocaine on the development rate of Boettcherisca peregrina (Diptera: Sarcophagidae). Journal o f Medical Entomology 1989; 26:91-93.

Goff, M.L., W.A. Brown, K.A. Hewadikaram, and A.I. Omori. Effect of heroin in decomposing tissues on the developmental rate of Boettcherisca peregrina (Diptera: Sarcohpagidae) and implications of the effect on estimation of postmortem interval using arthropod development patterns. Journal o f Forensic Science 1991; 36:537-542.

Goff, M.L., W.A. Brown, and A.I. Omori. Preliminary observations on the effects of methamphetamine in decomposing tissues on the development of Parasarcophaga ruficomis (Diptera: Sarcophagidae) and implications of this effect to estimation of postmortem interval. Journal o f Forensic Science 1992; 37:867-872.

Goff, M.L., W.A. Brown, A.I. Omori, and D.A. LaPointe Preliminary observations on the effects of amitryptyline in decomposing tissues on the development of Parasarcophaga ruficomis (Diptera: Sarcophagidae) and implications of this effect to estimation of postmortem interval. Journal o f Forensic Science 1993; 38:316-322. Preliminary observations on the effects of phencyclidine in decomposing tissues on the development of Parasarcophaga ruficomis (Diptera: Sarcophagidae). Journal o f Forensic Science 1994; 39:123-128.

Goff, M.L. and W.D. Lord. Entomotoxicology. A new area for forensic investigation. American Journal o f Forensic and Medical Pathology 1994; 15:51-57.

Greenberg, Bernard Flies and disease. VolII. Princeton University Press, 1973. A manual of forensic entomology (book review). Journal o f the New York Entomological Society 1988; 96:489-491. Nocturnal behavior of blow flies (Dipter:Caliphoridae). Journal of Medical Entomology 1990;25:199-200. Flies as forensic indicators. Journal o f Medical Entomology 1991; 28:565-577.

Greenberg, Bernard and Devinder Singh. Species Identification of Calliphorid (Diptera) Eggs. Journal o f Medical Entomology 1995; 32(l):21-26.

68 Groth, U.V. and H. Reissmuller. Beziehungen synanthroper Fliegen zu Kleintierleichen I. Teil: Methodik, Vorund Hauptversuche-Angew. Parasitol 1973; 14:83-100.

Hackman, W. Studies on the dipterous fauna in burrows of voles (Microtus, Clethrionomys) in Finland. Acta Zoologica Fennica 1963; 102:1-64.

Hall, D.G. The blowflies o f North America. Thomas Say Foundation, Lafayett, Indiana, 1947.

Hanski, Illka and Seppo Kuusela. An experiment on competition and diversity in the carrion fly community. Annales Zoologici Fennici 1977; 43(4): 108-114.

Haskell, Neal H. Entomological Collection Techniques at Autopsy and for Specific Environments. In Entomology and Death: Procedural Eds. Guide, E. Paul Catts and Neal H. Haskell, South Carolina: Joyce's Print Shop, Inc., 1990.

Haskell, Neal H. and Ralph E. Williams. Collection of Entomological Evidence at the Death Scene. In Entomology and Death: Procedural Eds. Guide, E. Paul Catts and Neal H. Haskell, South Carolina: Joyce's Print Shop, Inc., 1990.

Hewadikaram, K.A. and M.L. Goff. Effect of carcass size on rate of decomposition and arthropod succession patterns. American Journal o f Forensic Medicine and Pathology 1991; 12(3):235-240.

Hitchcock, Leo C. and Cronquist, Arthur. Flora o f the pacific northwest. University of Washington Press, Seattle, 1973.

Howden, A.T. The Succession o f Beetles on Carrion. Unpublished dissertation for degree of MS, North Carolina State College, Raleigh, 1950.

Introna, Francesco, Carlo Pietro Campoibasso, and Aldo Di Fazio. Three Case Studies in Forensic Entomology from Southern Italy. Journal o f Forensic Sciences 1998; 43(1):210-214.

Jiron, L.F. and V.M. Cartin. Insect succession in the decomposition of a mammal in Costa Rica. Journal o f the New York Entomological 1981; Society 89:158.

Johnson, M.D. Season and microserai variations in the insect population on carrion. American Midland Naturalist 1975; 93:79-90.

Kamal, A.S. Comparative study of thirteen species of sarcosaprophagous Calliphoridae and Sarcophagidae (Diptera). I. Bionomics. Annals o f the Entomological Society for America 1958; 51:261-271.

Kneidel, K.A. Competition and disturbance in communities of carrion-breeding Diptera. Journal o f Animal Ecology 1984; 53:849-865.

69 Komar, Debra and Owen Beattie. Postmortem Insect Activity may Mimic Perimortem Sexual Assault Clothing Patterns. Journal of Forensic Sciences 1998; 43(4):792-796.

Kuusela, Seppo and Ilkka Hanski. The structure of carrion fly communities: the size and the type of carrion. Holarctic Ecology 1982; 5:337-348.

LeClercq M. On the entomofauna of a wild boar's cadaver. Bulletin & Annales de la Societe Royale Beige d'Entomologie 1997; 132(4):417-422 (In French).

Lord, W.D. Case Histories of the use of insects in investigations. In Entomology & Death: A Procedural Guide, edited by P.E. Catts and N.H. Haskell, pp.9-37. Joyce's Print Shop, Clemson, 1990.

Lord, W.D. and John F. Burger. Collection and preservation of forensically important entomological materials. Journal o f Forensic Sciences 1983; 28(4):936-944.

Lord, W.D. and J.R. Stevenson. Directory of Fomesic Entomologists. 2nd edition. Am. Reg. Prof. Entomol., Washington D.C., 1986.

Malloch, John R. A Preliminary Classification of Diptera, Exclusive of Pupipara, Based upon Larval and Pupal Characters, with keys to Imagines in Certain Families. Part I. Bulletin o f the Illinois State Laboratory o f Natural 1917; History Vol. XII Article III.

Maus, Frank. Personal Communication. Forest Manager, Lubrecht Experimental Forest, The University of Montana School of Forestry, Greenough, MT, 1999.

Meek, C.L., M.D. Andis, and C.S. Andrews. Role of the Entomologist in Forensic Pathology, Including a Selected Bibliography. Bibliographies o f the Entomological Society o f America 1983; 1:1-10.

Megnin, P. La Faune des Tombeaux. Compte Rendu Hebdomadaire des Seances de VAcademie des Sciences, Paris, 1988; 105:948-951. La Faune des cadavres: application de l'entomologie a la medicine legale. Encyclopedic Scientifique des Aide-Memoire. Masson, Gauthier-Villars Paris, et Fils, 1894; 1.

Motter, Murray Galt. A contribution to the study of the fauna of the grave. A study of one hundred and fifty disinterments, with some additional experimental observations. The New York Entomological Society 1898; 6(4):203-230.

Nishida, K. Experimental studies on the estimation of postmortem intervals by means of fly larvae infesting human cadavers. Japanese Journal o f Legal Medicine 1984; 38:24- 42.

70 Nuorteva, P. Histerid beetles as predators of blowflies (Diptera, Calliphoridae) in Finland. Annales Zoologici Fennici 1970; 7:195-198. Age determination of a blood stain in a decaying shirt by entomological means. Forensic Science 1974; 3: 89-94. Sarcosaprophagous insects as forensic indicators, vol 2, pp. 1072-1095. In Forensic medicine: a study in trauma and environmental Edited hazards. by C. G. Tedeschi, W.G. Eckert and L.G. Tedeschi, Saunders, Philadelphia, 1977. Empty puparia of Phormia terraenova R.D. (Diptera, Calliphoridae) as forensic indicators. Annales Entomologici Fennici 1987; 53:53-56.

Nuorteva, P., M. Isokoski, and K. Laiho. Studies on the possibilities of using blowflies (Diptera) as medico-legal indicators in Finland. Annales Entomologici Fennici 1967; 33:217-225.

Okely, E.F. Descriptions of the puparia of twenty three British species of Sphaeroceridae (Diptera; Acalyptratae). Transactions o f the Royal Entomological Society o f London 1974; 126:41-56.

Oldroyd, Harold and Kenneth G.V. Smith. Eggs and Larvae of Flies. In insects and other arthropods o f medical importance, K.G.V. Smith editor, British Museum of Natural History, London, 1973: 289-323.

Palmer, D.H. Partitioning o f the carrion resource by sympatric Calliphoridae (Diptera) near Melbourne. PhD Thesis, La Trobe University, Melbourne, Australia, 1980.

Patrican, L.A. and R. Vaidyanathan. Arthropod succession in rats euthanized with carbon dioxide and sodium pentobarbital. Journal o f the New York Entomological Society 1995; 103(2): 197-207.

Payne, J.A. A summer carrion study of the baby pig Sus scrofa Linnaeus. Ecology 1965; 46:592-602.

Payne, Jerry A., Frank W. Mead, and Edwin W. King. Hemiptera associated with pig carrion. Annals o f the Entomological Society of America1968; 61(3):565-567.

Payne, J.A., and E.W. King Coleoptera associated with pig carrion. Entomologist's Monthly Magazine 1970; 105:224-232. Insect succession and decomposition of pig carcassses in water. Journal o f the Georgia Entomological Society 1972; 7(3): 153-162.

Peet, Robert K. Forests of the Rocky Mountians. In North American terrestrial vegetation. Cambridge University Press, Cambridge. Edited by Barbour, Michael G. and William Dwight Billings., 1988.

71 Prichard, J.G., P.D. Kossoris, R.A. Leibovitch, L.D. Robertsoon and F.W. Lovell. Implications of trombiculid mite bites: report of case and submission of evidence in a murder trial. Journal o f Forensic Sciences 1986; 31:301 -306.

Putman, R. J. Flow of energy and organic matter from a carcass during decomposition. Oikos 1978; 31: 58-68.

Ratcliffe, B.C. The natural history of Necrodes surinamensis. Transactions o f the American Entomological Society 1972; 98:359-410.

Redi, F. Esperienze intomo alia generazione degl' insetti. Insegna della Stella, Florence, 1668.

Reed, H.B. A study of dog carcass communities in Tennessee, with special reference to the insects. The American Midland Naturalist 1958; 59:213-245.

Reiter, C. and G. Wollenek. Zur Artbestimmung der Puparien forenisisch bedeutsamer Schmeissfliegen. Z. Rechtsmed 1983; 91:61-69.

Renfrew, C. and P. Bahn. Archaeology: Theories, method, and practice. Thames and Hudson Inc., New York, New York, 1991.

Rueda, Leopoldo M., Pyong-Ui, Roh, and Jang Leun Ryu. Pupal parasitoids (Hymenoptera: Pteromalidae) of filth flies (Diptera: Muscidae, Calliphoridae) breeding in refuse and poultry and livestock manure in South Korea. Journal o f Medical Entomology 1997; 34(l):82-85.

Schoenly, K. and Reid, W. Community structure of carrion arthropods in the Chihuahuan Desert. Journal o f Arid Environment 1983; 6:253-263.

Shean, Blair S., Lynn Messinger and Mark Papworth. Observations of differential decomposition on sun exposed vs. shaded pig carrion in coastal Washington state. Journal o f Forensic Sciences 1993; 38(4):938-949.

Smith, K.G.V. A Manual o f Forensic Entomology. Cornell University Press, Cornell University Press, New York, 1986.

Springett, B.P. Aspects of the relationship between burying beetles, Necrophorus SPP. And the mite, Poecilochirus Necrophori Vitz. Journal o f Animal Ecology 1968; 37:417- 424.

Tantawi,T.I. and E.M. El-Kady. Identification of third instar larvae of forensically important flies (Diptera: Calliphoridae, Sarcophagidae and Muscidae) in Alexandria, Egypt. Journal ofthe Egytian German Society o f Zoology1997; 23(E): 1-20.

72 Tantawi, T.I., El-Kady, E.M., Greenberg, B., and El-Ghaffar, H.A. Arthropod succession on exposed rabbit carrion in Alexandria, Egypt. Journal o f Medical Entomology 1996; 33:566-580.

Temeny, Tiffany T. Estimation of time since death in humans using mature pigs. Masters Thesis, University of Montana, Missoula, 1997.

Tullis, K., and M.L. Goff. Arthropod succession in exposed carrion in a tropical rainforest on Oahu, Hawaii. Journal o f Medical Entomology 1986; 24: 332-339.

Utsumi, K. Studies on arthropods congregating in animal carcasses, with regard to the estimation of postmortem interval. Ochanomizu Medical Ann. (Tokyo) 1958; 7:202.

Vincent, C., D.K. Kevan, M. Leclercq, and C.L. Meek. A bibliography of forensic entomology. Journal o f Medical Entomology 1985; 22: 212-219.

Webb, J.P.,Jr., R.B. Loomis, M.B. Madon, S.G. Bennett and G.E. Greene. The chigger species Eutrombicula belkini Gould (Acari: Trombiculidae) as a forensic tool in a homicide investigation in Ventura County, California. Bulletin for the Society o f Vector Ecology 1983; 8:141-146.

Williams, H. and A.M.M. Richardson. Growth energetics in relation to temperature for larvae of four species of necrophagous flies (Diptera: Calliphoridae). Australian Journal o f Ecology 1984; 9:141-152.

73