Calliphora vicina (Diptera: Calliphoridae) collected from a human corpse above 3400 m in elevation

In the summer of 2005 an adult hiker was killed in a fall in Rocky Mountain National Park, Colorado (U.S.A.). His body was subsequently colonized by the blowfly Calliphora vicina (Diptera: Calliphoridae). The body was discovered 9 days later at an elevation of 3475 m (11,400 feet). This is the first report of C. vicina on a human corpse at this elevation in Colorado. This paper provides additional confirmation of this fly colonizing a human corpse at high elevations in Colorado.     

A test for carrion fly full siblings: a tool for detecting postmortem relocation of a corpse

We propose a genetic test for full sibship for a pair of carrion flies that could reveal the postmortem relocation of a corpse. A carrion fly larva is sometimes left behind when a corpse is moved. The discovery of full sibling larvae of approximately the same developmental stage at two locations would strongly suggest that a corpse was moved between those two sites. Distributions of pairwise comparisons of relatedness (R) coefficients were generated using amplified fragment length polymorphism profiles for nine samples of laboratory-generated full siblings as well as for a reference sample of nonfull sibling Phormia regina (Diptera: Calliphoridae). The mean relative R coefficient, a pairwise measure of the proportion of shared alleles, was 0.479 (±0.289 SD) for full siblings, close to the theoretical expectation of 0.5. A likelihood ratio (LR) test was based on observed distributions of R. R >0.55 corresponded to an LR >1000 favoring full sibship for that pair of individuals.     

A forensic entomology case from the Amazon rain forest of Brazil

The first case of application of forensic entomology in the Brazilian Amazonia is described. The corpses of 26 men were found in the rainforest in Rondonia State, Brazil. Fly larvae collected on the bodies during autopsy were identified as Paralucilia fulvinota (Diptera, Calliphoridae). No data or specimens were collected at the crime scene. At the laboratory, the larvae developed into pupae in 58 h and into adults in 110.5 h. The total development time for P. fulvinota was measured in field experiments inside the forest. The age of the larvae when collected from the bodies was estimated as the difference between the time required for them to become adults and the total development time for this species. The estimated age of the maggots and the minimum postmortem interval was 5.7 days.     

Developmental Times of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) at Constant Temperatures and Applications in Forensic Entomology

The characteristic life stages of infesting blowflies (Calliphoridae) such as Chrysomya megacephala (Fabricius) are powerful evidence for estimating the death time of a corpse, but an established reference of developmental times for local blowfly species is required. We determined the developmental rates of C. megacephala from southwest China at seven constant temperatures (16–34°C). Isomegalen and isomorphen diagrams were constructed based on the larval length and time for each developmental event (first ecdysis, second ecdysis, wandering, pupariation, and eclosion), at each temperature. A thermal summation model was constructed by estimating the developmental threshold temperature D₀ and the thermal summation constant K. The thermal summation model indicated that, for complete development from egg hatching to eclosion, D₀ = 9.07 ± 0.54°C and K = 3991.07 ± 187.26 h °C. This reference can increase the accuracy of estimations of postmortem intervals in China by predicting the growth of C. megacephala.     

The black soldier-fly, Hermetia illucens (Diptera, Stratiomyidae), used to estimate the postmortem interval in a case in Amapá State, Brazil

The black soldier-fly (Hermetia illucens) is a generalist detritivore which is commonly present in corpses in later stages of decomposition and may be useful in forensic entomology. This paper describes the estimation of the postmortem interval (PMI) based on the life cycle of the black soldier-fly in a case in northern Brazil. A male child was abducted from his home and 42 days later his corpse was found in an advanced stage of decay. Two black soldier-fly larvae were found associated with the body. The larvae emerged as adults after 25-26 days. Considering the development cycle of H. illucens, the date of oviposition was estimated as 24-25 days after abduction. Since H. illucens usually (but not always) colonizes corpses in more advanced stages of decay, this estimate is consistent with the hypothesis that the child was killed immediately after abduction.     

Effects of Ketamine on the Development of forensically important Blowfly Chrysomya megacephala (F.) (Diptera: Calliphoridae) and its Forensic Relevance

This study investigated effects of ketamine on the development of Chrysomya Megacephala (Diptera: Calliphoridae) at three different temperatures. Larvae of the C. Megacephala were exposed to different concentrations of drugs and temperatures. The larval lengths, weights, and developmental durations of each stage were observed. This study demonstrated that ketamine, low temperature, and their synergistic action significantly suppressed the development of C. Megacephala (p < 0.001). The time that the larvae in all the treatments achieved the maximum length/weight was significantly delayed (p < 0.05), and that resulted in prolonged duration of larval and prepupal stages especially at low temperature. However, no linear correlations were discovered between ketamine concentration and growth rate of larval length/weight.     

First Report of Blaesoxipha plinthopyga (Diptera: Sarcophagidae) from a Human Corpse in the U.S.A. and a New State Geographic Record Based on Specimen Genotype

Carrion flies in the taxonomic family Sarcophagidae are often recovered from a human corpse. However, because such specimens are difficult to identify, the forensic literature on this taxon is quite limited compared with that of the commonly employed Calliphoridae. Faced with a sarcophagid larva that could not be identified microscopically from a death investigation in the state of Idaho, we generated cytochrome oxidase one DNA sequence data from the specimen. Comparison to a reference data set of forensically significant sarcophagids from Canada and the U.S.A. confirmed that this was the first discovery of Blaesoxipha plinthopyga in a human corpse in the U.S.A. and the first record of this species in Idaho. Because B. plinthopyga occurs from the Northern U.S.A. to the Neotropics, it is potentially useful for estimating time since death at many locations.     

Potential Use of Hydrocarbons for Aging Lucilia sericata Blowfly Larvae to Establish the Postmortem Interval

Previous studies on Diptera have shown the potential for the use of cuticular hydrocarbons' analysis in the determination of larval age and hence the postmortem interval (PMI) for an associated cadaver. In this work, hydrocarbon compounds, extracted daily until pupation from the cuticle of the blowfly Lucilia sericata (Diptera: Calliphoridae), have been analyzed using gas chromatography–mass spectrometry (GC–MS). The results show distinguishing features within the hydrocarbon profile over the period of the larvae life cycle, with significant chemical changes occurring from the younger larvae to the postfeeding larvae. Further interpretation of the chromatograms using principal component analysis revealed a strong correlation between the magnitudes of particular principal components and time. This outcome suggests that, under the conditions of this study, the cuticular hydrocarbons evolve in a systematic fashion with time, thus supporting the potential for GC–MS analysis as a tool for establishing PMI where such a species is present.     

Human Cutaneous Myiasis by the Australian Sheep Blowfly,Lucilia cuprina (Diptera: Calliphoridae), in Oklahoma

This case report describes an incident of myiasis in Oklahoma City, Oklahoma. Paramedics treated a 53‐year‐old man in the field when he suffered cardiac arrest. He was intubated by the paramedics and transported to an emergency room, where he received two stents and a balloon pump. He was found to have a GCS of three and remained in the hospital for 7 days before passing away. After his death, the breathing tube was removed, revealing the presence of several maggots. On closer inspection, the dipteran larvae were found in both the oral and nasal cavities. Four of these larvae were reared to adulthood and identified as Lucilia cuprina (Weidemann) (Diptera: Calliphoridae), commonly known as the Australian sheep blowfly, by the Biology Department of the University of Oklahoma. Based on the fly's life cycle and rate of development, the infestation is suspected to be hospital acquired.     

Determination of Calliphoridae (Diptera) Fauna and Seasonal Distribution on Carrion in Ankara Province*

Abstract: This study was conducted from March 2006 to 2007. The aim of the study was to determine the forensically significant Calliphoridae (Diptera) species and their seasonal distribution in Ankara province at Beytepe Campus. Pig carcasses were killed by a veterinary using pentobarbital sodium with intervals from 2 to 3 weeks to a month. Samples were collected from the carcass everyday in the morning, in the afternoon, and at sunset. Temperature and parameters of weather, such as cloudy, sunny, rainy, and snowy, were recorded at each visit. Phaenicia sericata (Meigen), Chrysomya albiceps (Wiedemann), Calliphora vomitoria (Linnaeus), and Calliphora vicina (Robineau‐Desvoidy) species, which belong to Calliphoridae family, were collected. It was shown that the seasonal distribution of the collected species was different from each other. The species were determined on carcass between the following times; P. sericata, from April to November; C. albiceps, from May to November; Ca. vomitoria, from February to June and from September to December; and Ca. vicina, from February to May and in June and September. Calliphoridae fauna and its annual seasonal distribution in Turkey on 12 pig carcasses were reported for the first time in this study.     

Three species of insects collected from an adult human corpse above 3300 m in elevation: a review of a case from Colorado

We report on the colonization of an adult human corpse by three insect species at 3350 m (11,000 ft) in elevation. The adult silphid Thanatophilus coloradensis (Wickham), adults of the blow fly Calliphora coloradensis (Hough), and larvae and adults of Lucilia silvarum (Meigen) were all collected from the victim's body which had been wrapped in plastic. The victim was found in late June in the Rocky Mountains of Colorado. This paper provides additional confirmation of the taxa utilizing a human corpse at high elevations in Colorado.     

Fluorescence Imaging of Posterior Spiracles from Second and Third Instars of Forensically Important Chrysomya rufifacies (Diptera: Calliphoridae),

Entomological protocols for aging blowfly (Diptera: Calliphoridae) larvae to estimate the time of colonization (TOC) are commonly used to assist in death investigations. While the methodologies for analyzing fly larvae differ, most rely on light microscopy, genetic analysis, or, more rarely, electron microscopy. This pilot study sought to improve resolution of larval stage in the forensically important blowfly Chrysomya rufifacies using high‐content fluorescence microscopy and biochemical measures of developmental marker proteins. We established fixation and mounting protocols, defined a set of measurable morphometric criteria and captured developmental transitions of 2nd instar to 3rd instar using both fluorescence microscopy and anti‐ecdysone receptor Western blot analysis. The data show that these instars can be distinguished on the basis of robust, nonbleaching, autofluorescence of larval posterior spiracles. High‐content imaging techniques using confocal microscopy, combined with morphometric and biochemical techniques, may therefore aid forensic entomologists in estimating TOC.     

Growth of Chrysomya megacephala (Fabricius) Maggots in a Morgue Cooler*

Abstract: In estimating the postmortem interval (PMI) using maggots obtained during autopsy, the forensic entomologist makes decisions regarding the effects of low‐temperature storage of the body on the insects. In this case report, a corpse was found in an abandoned house in the residential area of Bukit Mertajam, Penang, Malaysia. The maggots were found to be alive inside the mouth of the deceased although the corpse had been in the morgue cooler for 12 days. The maggots were reared and identified as Chrysomya megacephala (Fabricius). The emerged adult flies were kept as a stock colony, and the duration of development under the indoor fluctuating temperature regime was studied. The total duration of developmental process of this species was 9.5 ± 0.5 days, and the PMI estimated was 3.2 ± 0.6 days. This case report demonstrates the survival of Ch. megacephala maggots for 12 days and their growth inside the morgue cooler.     

The Use of the Developmental Rate of the Aquatic Midge Chironomus riparius (Diptera,Chironomidae) in the Assessment of the Postsubmersion Interval

Nonbiting midges (Diptera, Chironomidae) are the most abundant members of the fauna associated with submerged carcasses, but their use in the medicolegal context is very restricted because of their complex ontogeny. In this case, the corpse of a woman was recovered in late spring from a river in Granada (Iberian Peninsula). It showed obvious signs of long permanence in the aquatic environment and, along with pulmonary and microscopical analyses, led to the conclusion that the cause of death was drowning. Several larvae‐like specimens were sampled from the scalp and later identified by morphological external features as IV instar larvae of Chironomus riparius Meigen, 1804 (Diptera, Chironomidae). Sequencing of cytochrome oxidase subunit I was performed to confirm the identification. The knowledge of the biology of C. riparius at low temperatures was critical to assess a postsubmersion interval of 16–17 days.     

Toxicology and fly larvae on a putrefied cadaver

Toxicological analyses on a putrefied cadaver are sometimes difficult to achieve, due to the absence of blood and/or urine. In this study, morphine and phenobarbital were simultaneously identified and assayed in several tissues of a putrefied cadaver and in the fly larvae of Calliphoridae found on the corpse.     

Influence of substrate tissue type on larval growth in Calliphora augur and Lucilia cuprina (Diptera: Calliphoridae)

The size of fly larvae is an important variable in the use of these insects to estimate postmortem interval. Furthermore, the nutritional intake of larvae is likely to vary subject to the part of a corpse on which they are feeding. A study was therefore conducted to investigate the effect of type of food substrate on larval growth in two species of forensically important Australian blowflies. After collection on sheep's liver in the laboratory, different groups of larvae of Lucilia cuprina (Wiedemann) and Calliphora augur (Fabricius) were grown on sheep's liver, meat, and brains, and their body lengths compared. Results indicated that the development of larvae fed sheep's liver was adversely affected compared with larvae fed meat and brain; they moulted later, reached maximum length more slowly and sometimes produced significantly smaller pupae. These findings, similar to those of another recent study, have obvious implications for postmortem interval determinations. Estimates may be considerably skewed if the site of collection of larvae at a death scene contains tissue types different to those used in reference experiments. We therefore recommend caution in forensic analyses that interpret crime scene data using developmental studies performed with a single type of larval food substrate.     

Eristalinus arvorum (Fabricius, 1787) (Diptera: Syrphidae) in Human Skull: A New Fly Species of Forensic Importance

A body of an unknown adult female was found within a shallow burial ground in Malaysia whereas the skull was exposed and visible on the ground. During autopsy examination, nine insect larvae were recovered from the interior of the human skull and subsequently preserved in 70% ethanol. The larvae were greyish in appearance, each with a posterior elongated breathing tube. A week after the autopsy, more larvae were collected at the burial site, and some of them were reared into adults. Adult specimens and larvae from the skull and from the burial site were sequenced to obtain DNA barcodes. Results showed all adult flies reared from the burial site, as well as the larvae collected from the skull were identified as Eristalinus arvorum (Fabricius, 1787) (Diptera: Syrphidae). Here, we report the colonization of E. arvorum larvae on a human corpse for the first time.     

Analysis of the Effect of Cyclophosphamide and Methotrexate on Chrysomya megacephala (Diptera: Calliphoridae),

Forensic entomotoxicology investigates the effects of chemical substances in the development of scavenger insects and the reflection on estimating the minimum postmortem interval (PMI). To evaluate the impact of the chemotherapeutic drugs cyclophosphamide (CF) and methotrexate (MTX) on the postembryonic development of Chrysomya megacephala, controlled experiments were performed by rearing the larvae on spiked minced beef with different concentrations of these drugs. The results indicated that CF significantly decreased their developmental rate up to 28 h but had no significant effect on larval and adult sizes, survival rate, and sex ratio, whereas MTX decreased larval and adult sizes, survival rate, and there was a deviation in the expected sex ratio toward females in MTX‐exposed larvae but had no significant impact on developmental rate. These negative interference factors should be considered in cases of suspected death of people that have undergone chemotherapy.     

Diurnal and Nocturnal Flight Activity of Blow Flies (Diptera: Calliphoridae) in a Rainforest Fragment in Brazil: Implications for the Colonization of Homicide Victims

Nocturnal flight of blow flies (Diptera: Calliphoridae) is a controversial issue in forensic entomology. We performed two field experiments to investigate the diurnal and nocturnal activity of six blow fly species in a rainforest fragment in Brazil. Initially, nocturnal (17:30–05:30) versus diurnal (05:30–17:30) flight activity was investigated. Only 3.9% of adults were collected at night, mostly the native species Mesembrinella bicolor, and nocturnal oviposition did not occur. In the second experiment, collection of adults took place at the following intervals: 05:30–08:30, 08:30–11:30, 11:30–14:30, and 14:30–17:30. The proportions of adults did not differ significantly among the four diurnal intervals, except for Hemilucilia segmentaria, which was captured more frequently in the early morning. Calliphoridae has predominantly diurnal behavior, not laying eggs in darkness. The association of the native species M. bicolor, Hemilucilia semidiaphana, and H. segmentaria to forested areas reinforces the forensic relevance of data on their flight pattern.     

Detection of Pb,Ba, and Sb in Cadaveric Maggots and Pupae by ICP-MS*

The concentrations of lead (Pb), barium (Ba), and antimony (Sb), characteristic of GSR, were determined in soil sediments and immature (larvae) of cadaveric flies of the family Calliphoridae, by inductively coupled plasma mass spectrometry (ICP-MS). This research refers to a case study from two real crime scenes in which the corpses were in an advanced state of decomposition. In case 1, the victim had holes similar to gunshot wounds, and in case 2, there was no evidence of perforations in the corpse. Soil sediment collection was performed at three different points of the terrain, at a minimum distance of 10 m from the corpse, for cases 1 and 2. In relation to the collection of immatures, larvae were collected in regions of the mouth, nose, and orifices similar to the entry of firearms projectile into the body, for case 1, and collection of larvae and pupae, located on the body and underneath it, for case 2. It was possible to detect and quantify the three elements of interest (Pb, Ba, and Sb) by ICP-MS in both sediment and cadaveric larvae. Concentrations of 4.44, 8.74, and 0.08 μg/g were obtained for Pb, Ba, and Sb, respectively, in the soil for case 1. For the case 2, the concentrations in Pb, Ba, and Sb were from 16.34 to 26.02 μg/g; from 32.64 to 57.97 μg/g and from 0.042 to 0.30 μg/g, respectively. In the larvae, Pb, Ba, and Sb were quantified in cases 1 and 2 with a concentration of 6.28 and 1.78 μg/g for Pb, 1.49 and 2.94 μg/g for Ba, 0.50 μg/g and <LD for Sb, respectively. These new results present the detection of characteristic elements of GSR in cadaveric larvae in humans in a real crime scene, besides highlighting the importance of the study of immature flies, using the ICP-MS technique in forensic analysis.