Necrophagous Flies Research Articles

Evaluation of Larval Therapy Compared to Antibiotic Therapy in the Treatment of Skin Wounds in Rabbits.

Larval therapy (LT) is a therapeutic modality that uses larvae of necrophagous flies for the treatment of wounds. The use of this therapy presents several benefits, due to the action of the larvae that remove necrotic tissue selectively, exercising antimicrobial action, and promoting healing. There are situations in which LT proves to be the only or the best alternative for the patient, such as wounds infected by multidrug-resistant microorganisms or when treatment difficulties may lead to an indication for amputation. The purpose of this study was to compare the efficiency of LT, using larvae of Chrysomya megacephala, with that of antibiotic therapy in the treatment of cutaneous wounds infected with Pseudomonas aeruginosa. Twenty-four rabbits were used to perform the experiment, distributed in four groups: group 1, induced wound without bacterial infection; group 2, induced wound with bacterial infection; group 3, induced wound with bacterial infection and antibiotic therapy; group 4, induced wound with bacterial infection and LT. The macroscopic, microscopic, and statistical analyses indicated that LT was as effective as antibiotic therapy in wound healing.

COⅠ and 16S rDNA Sequence Identification of Common Necrophagous Flies in Fujian Province

Objective To identify the species of common necrophagous flies in Fujian Province by gene fragment sequences of mitochondrial cytochrome c oxidase subunit Ⅰ （COⅠ） and 16S ribosomal deoxyribonucleic acid （16S rDNA）, and to explore the identification efficacy of these two molecular markers. Methods In total 22 common necrophagous flies were collected from the death scenes in 9 different regions in Fujian Province and DNA was extracted from the flies after morphological identification. The gene fragments of COⅠ and 16S rDNA were amplified and sequenced. All the sequences were uploaded to GeneBank and BLAST and MEGA 10.0 software were used to perform sequence alignment, homology analysis and intraspecific and interspecific genetic distance analysis. The phylogenetic trees of DNA fragment sequences of COⅠ and 16S rDNA of common necrophagous flies in Fujian Province were established by unweighted pair-group method with arithmetic means （UPGMA）, respectively. Results The flies were classified into 6 species, 5 genera and 3 families by morphological identification. The results of gene sequence analysis showed that the average number of interspecific and intraspecific genetic distance of 16S rDNA ranged from 1.8% to 8.9% and 0.0% to 2.4%, respectively. The average number of interspecific and intraspecific genetic distance of COⅠ ranged from 7.2% to 13.6% and 0.0% to 6.3%, respectively. Conclusion The gene sequences of COⅠ and 16S rDNA can accurately identify the species of different necrophagous flies, and 16S rDNA showed higher value in species identification of common calliphoridae necrophagous flies in Fujian Province.

The Application of Mitochondrial COI Gene-Based Molecular Identification of Forensically Important Scuttle Flies (Diptera: Phoridae) in Korea.

Phoridae are a family of necrophagous flies commonly found in indoor death scene. They account for approximately 19.7% of the entomofauna in human cadavers in Korea. Additionally, this taxon is an indicator of indoor hygiene, and these flies appear in environments where access by other necrophagous insects is difficult, such as enclosed rooms. Thus, they are likely to be used as forensic evidence. Despite their importance in forensic investigations and environmental hygiene, detailed studies on the taxonomy and molecular barcoding for this family are scarce, including in Korea. Because accurate taxonomic information regarding necrophagous insects collected from a death-related scene is essential during medicolegal investigations, molecular barcoding data could be useful as well as reliable. In this paper, full-length nucleotide sequences of genes coding for the cytochrome c oxidase subunit I (COI) in 79 Phoridae larvae collected from 20 medicolegal autopsy cases in Korea were phylogenetically analyzed by comparing their sequences to the foreign barcoding data of Phoridae. Six mitochondrial haplogroups were identified, which two of them matched to foreign Phoridae fly species haplotypes, Megaselia scalaris (Loew, 1866) and M. spiracularis Schmitz 1938. Taxonomies of five other haplogroups, with nucleotide distances ranging from 1.68% to 2.26% from the M. scalaris group, could not be confirmed solely based on the molecular barcoding data. Further research should be performed to determine whether these five haplogroups are diverged conspecifics of M. scalaris or a closely related sister cryptic species of M. scalaris.

Cuticular hydrocarbons as a tool for determining the age of Chrysomya rufifacies (Diptera: Calliphoridae) larvae.

Calliphoridae are one of the most important insect groups encountered as evidence collected from a crime scene. Age determination of the immature stages of these necrophagous flies is an important step toward estimating the time of colonization and inferring a minimum postmortem interval (PMImin ) in most instances. To determine if the cuticular hydrocarbons could be used to establish whether the development stages yield characteristics profiles, allowing for age estimation, hydrocarbons were extracted from 1st and 2nd, as well as feeding and post-feeding 3rd instar Chrysomya rufifacies, the hairy maggot blow fly. Extracted hydrocarbons were analyzed using gas chromatography coupled to mass spectrometry with the aim to investigate the changes in chemical profiles of each larval stage. A total of 23 compounds were identified with most of them being alkanes (65%) with carbon chain lengths of 9-33 carbons, alkenes (18%), and methyl-branched alkanes (17%). All the hydrocarbons except pentadecane (C15), hexadecane (C16), and nonacosane (C29) showed significant differences in their expression throughout larval development. For 1st instars, nonane was the most abundant (17% of the total hydrocarbons content) compound. Accounting for 11% and 10% of the cuticular hydrocarbons, tricosane and pentacosane, respectively, were the notable hydrocarbons in 2nd instars. For post-feeding 3rd instars, hentriacontane and tritriacontane were present with relative abundances 18% and 15%, respectively. On average, there was a shift from low to high molecular weight hydrocarbons as the larvae aged. These results indicate the change in hydrocarbons makeup as larvae age and could potentially be used to determine the age of immature C.rufifacies and hence aid in PMImin estimations. However, future research is needed to validate these results under natural conditions in the field.

Breaking the silence: how shedding light on the bone-skipper fly Thyreophora cynophila (Diptera: Piophilidae) demonstrated it still has a large distribution area in the Pyrenees mountains, France.

BackgroundThyreophora cynophila (Panzer, 1798) is an iconic species of the European entomofauna. This winter-flying necrophagous fly was considered long extinct in Europe, before being discovered in Spain in 2010 and re-discovered in France in 2020, with a unique locality in Saint-Paul-de-Jarrat (Ariège, southern France).New informationAfter bringing this species to the attention of people that are active in nature during the winter, including hunters, skilled naturalists, nature lovers and professional naturalists, we gathered seven new occurrence data for this species at six locations on the French flanks of the Pyrenees mountains. Those data considerably extend the known distribution of the species in Europe and allows mapping the first approximate extent of occurrence for this species in France.

Carrion decomposition and assemblage of necrophagous dipterans associated with Terbufos (Organophosphate) intoxicated rat carcasses

Organophosphate use represents one of the main causes of intentional and accidental poisoning in Brazil. Because they induce alteration in carcass decomposition and succession of necrophagous Diptera, they are important investigative tool in forensic entomology for determining the postmortem interval (PMI) and cause of death. Thus, this work aimed to evaluate the influence of Terbufos, the most commonly used organophosphate for suicides in Rio Grande do Norte, Brazil, in the decomposition process and necrophagous fly assemblies in intoxicated rat carcasses. For this, 150g female Wistar rats received, by gavage, 200µL of Terbufos (5mg/kg or 10mg/kg) or distilled water (control). Approximately 30 minutes after administration, animals were euthanized and distributed in suspended traps to decompose under environmental conditions. Decomposition was monitored daily, with photographic record and collection of the visiting dipterofauna until the dry phase. Data show that the higher dose of Terbufos i) accelerates carcass decomposition in 24h; ii) decreases the species richness and abundance of scavengers flies when compared to the control; iii) changes the succession pattern, delaying the arrival of important species for the PMI estimate and iv) causes 8% mortality of the visiting dipterofauna. This work provides relevant information about dipteran assemblages and changes in the cadaver decomposition process associated with Terbufos intoxication, which may assist in future investigative processes with suspected organophosphate poisoning.

The upper thermal tolerance for a Texas population of the hairy maggot blow fly Chrysomya rufifacies Macquart (Diptera: Calliphoridae)

The hairy maggot blow fly (Chrysomya rufifacies: Macquart) is an invasive necrophagous fly found throughout the continental United States. Chrysomya rufifacies is of medical/veterinary, forensic, and ecological importance due to its ability to cause myiasis, colonise human remains, and displace native Diptera. However, little is known about their upper thermal tolerance, which could be used to better predict their invasion potential. We investigated the upper thermal tolerance of C. rufifacies exposed to different temperatures (20–45 °C), times (1–6 h), and nutrients (no food or water, water only, or a food‐water mixture) for both sexes and two age ranges (young = 6–8 days post pupal emergence; old = 9–11 days post pupal emergence). As temperature or duration increased, the probability of knockdown increased (0–100% at 20 and 45 °C and from 41 to 75% at 1 and 6 h), while the probability of survival decreased (99–2% at 20 and 45 °C and from 75 to 28% at 1 and 6 h). The availability of nutrients increased thermal tolerance at moderate temperatures (40 and 42 °C). Female flies were more thermally tolerant than males (probability of knockdown = 49% vs 58%; probability of survival = 58% vs 46%). Thermal tolerance did not differ by age. These data reveal details about the upper thermal tolerance for a single population of C. rufifacies, and suggest that environmental and organismal factors ought to be considered in order to make meaningful predictions about the invasion potential of C. rufifacies in North America.

The effects of cattle ranching on the communities of necrophagous flies (Diptera: Calliphoridae, Mesembrinellidae and Sarcophagidae) in Northeastern Brazil

Most of the ongoing decline in biodiversity, worldwide, is due to human activities, in particular, the expansion of agriculture. In this context, we highlight the need for studies of the taxonomic groups that can provide insights into the dynamics of the ecological communities facing anthropogenic impacts. For this, we evaluated the effects of the environmental changes caused by cattle ranching on five phytophysiognomies (Cerrado Savanna, Amazon Forest, Palm Forest, Marshland, and Mangrove) in the state of Maranhao, Brazil. We tested the hypothesis that the species composition, abundance, and richness of the families Calliphoridae, Mesembrinellidae, and Sarcophagidae (Diptera) are affected by ranching in each of the phytophysiognomies. Specimens were collected at 90 sites, including 45 anthropic sites (cattle ranches) and 45 preserved habitats, using traps baited with bovine lung. We collected 15,023 calliphorids (11 species), 10,772 sarcophagids (52 species), and 241 mesembrinellids (one species). The results indicated significant differences between anthropic and preserved habitats in the species composition, abundance, and richness of sarcophagids, in particular in the Amazon Forest, where the highest species richness was recorded in the anthropic environments. In the case of the calliphorids and mesembrinellids, by contrast, significant differences were found in species composition and abundance in only in four of the five phytophysiognomies analyzed (excluding the Cerrado Savanna), while species richness only varied in the palm forest and marshland. In all cases, lower values were recorded in the anthropic environments. These results indicate that the insects of the families Sarcophagidae, Calliphoridae, and Mesembrinellidae respond differentially to the anthropic activity (cattle ranching) and can be used to evaluate this type of anthropogenic impact systematically. In addition, the Amazon Forest was the phytophysiognomy most impacted by this activity in the Brazilian state of Maranhao.

Detection of benzodiazepines in decomposing rabbit tissues and certain necrophagic dipteran species of forensic importance.

The determination of benzodiazepines (carbamazepine and clobazam) in rabbit carcass tissues and larvae of three Calliphorid flies is described. After feeding the rabbits with lethal and toxic doses, samples of larvae and carcass tissues were studied. Residual drugs were determined using Ultra-high-performance liquid chromatography – quadrupole time-of-flight mass spectrometry (UHPLC/QTOF-MS). Benzodiazepines and its main active metabolites have been detected in the rabbit tissues at different retention times depending on the dosage used (lethal or toxic). A total of 1150 insects were collected and 800 larvae of the flies Chrysomya albiceps, Lucilia sericata and L. silvarum were used in the analysis. The presence of benzodiazepines in the rabbit tissues has been shown to typically affect the larval development cycle of the three necrophagous flies. Chrysomya albiceps larvae feed on drugs developed faster, while the development of L. sericata and L. silvarum larvae slowed. These results indicate that drugs have an impact on the life cycles of insects, which suggests that the presence of these substances is a factor that needs to be taken into account when estimating the post-mortem interval (PMI).

Characterization of Insect Stains Deposited by Calliphora vicina (Diptera: Calliphoridae) on Shirt Fabrics.

Despite the fact that necrophagous flies are known to alter bloodstains and create unique artifacts, no research has occurred to date that has examined the characteristics of insect stains on textiles or fabrics. This study represents the first effort to characterize artifacts produced by adult Calliphora vicina Robineau-Desvoidy deposited on a range of shirt fabrics that varied in type, color, orientation, and yarn tension. In general, artifact morphology on any type of fabric was distorted in comparison to those observed on smooth and/or nonporous surfaces in previous studies. Consequently, distinction of artifact type could only be made broadly as digestive artifacts and transfer patterns, in which the latter was predominantly detected as tarsal tracks. None of the artifacts displayed satellite stains typical of human bloodstains found on textiles. Wicking was evident on all fabrics but was most pronounced with dri wick and jersey knit polyester in comparison to cotton knit. Digestive artifacts on any colored fabric, but especially with green and yellow shirt samples, resembled the reported color, size, and morphology of bloodstains generated in laboratory studies on a range of fabrics. Unique digestive artifacts were also detected as small, black, and nearly spherical. These defecatory stains did not appear to wet or wick into any of the fabrics. Digestive artifacts and tarsal tracks differentially interacted with front face stitch loops of clothing fabrics to yield distinct stain patterns. The implications of these observations in reference to bloodstain pattern analysis at crime scenes are discussed.

Dynamic transcriptome profiling exploring cold tolerance in forensically important blow fly, Aldrichina grahami (Diptera: Calliphoridae)

BackgroundAldrichina grahami (Diptera: Calliphoridae) is a forensically important fly, which has been widely applied to practical legal investigations. Unlike other necrophagous flies, A. grahami exhibits cold tolerance which helps to maintain its activity during low-temperature months, when other species are usually not active. Hence, A. grahami is considered an important forensic insect especially in cold seasons. In this study, we aim to explore the molecular mechanisms of cold tolerance of A. grahami through transcriptome.ResultsWe collected eggs and larvae (first-instar, second-instar and third-instar) at three different temperatures (4 °C, 12 °C and 20 °C) and performed RNA-seq analyses. The differentially expressed genes (DEGs) associated with the cold-tolerance were screened out. The Venn analysis of DEGs from egg to third-instar larvae at three different temperatures showed there were 9 common genes. Candidate biological processes and genes were identified which refer to growth, and development of different temperatures, especially the chitin and cuticle metabolic process. The series-clusters showed crucial and unique trends when the temperature changed. Moreover, by comparing the results of growth and developmental transcriptomes from different temperatures, we found that DEGs belonging to the family of larval cuticle proteins (LCP), pupal cuticle protein (CUP), and heat shock proteins (HSP) have certain differences.ConclusionsThis study identified functional genes and showed differences in the expression pattern of diverse temperatures. The DEGs series-clusters with increasing or decreasing trends were analyzed which may play an important role in cold-tolerance. Moreover, the findings in LCP, CUP and HSP showed more possible modulations in a cold environment. This work will provide valuable information for the future investigation of the molecular mechanism of cold tolerance in A. grahami.

Back from the dead II: Thyreophora cynophila (Panzer, 1798) (Diptera: Piophilidae) resurfaces in France after a 183-year-long absence

SummaryLast seen in France in 1836 by Robineau-Desvoidy, the orange-headed necrophagous fly Thyreophora cynophila (Panzer, 1798) has long been considered as extinct. However, it was rediscovered in 2010 in Spain. We report its first sighting in France after a 183-year-long absence. In February 2019, four specimens were collected in southern France (Saint-Paul-de-Jarrat, Ariège), in the Pyrenees.

Green bottle flies (Calliphoridae, Luciliinae) of Ecuador: geographic distribution, checklist and DNA barcodes

ABSTRACT Green bottle flies (Diptera, Calliphoridae, Luciliinae) comprise a diverse and cosmopolitan taxon, known from at least 1,500 species. They have become crucial elements in forensic investigations, as they spend part of their life cycle in decaying remains. Here, we review the distribution of eleven Luciliinae species in Ecuador: the monotypic Blepharicnema and ten Lucilia species. We identified specimens using morphological characters. Additionally, we DNA barcoded 43 specimens from three species using 658bp segments of the standard Cytochrome Oxidase I (COI) mitochondrial gen. Molecular and morphological identifications presented high correspondence, suggesting COI barcodes are an efficient tool for the identification of these three green bottle flies species. Geographical records are biased towards the northern Andean region, particularly near to large urban settlements. We remark the value to applied forensic research of continuous sampling of necrophagous flies under a variety of habitats and crime conditions.

Effects of Methamphetamine on the Development and Its Determination in Aldrichina grahami (Diptera: Calliphoridae).

Age determination of necrophagous flies serves as an important tool for postmortem interval (PMI) estimation in forensic investigations. Drugs or toxins in cadavers may alter the developmental time of larvae, and lead to deviation in PMI estimation. Methamphetamine (MA), as one of the most abused psychostimulant drugs in Asia and North America, is often involved in forensic entomotoxicological cases. This study investigated the effects of MA (0, 45, 90, and 180 ng/mg) on the developmental rate, morphology, and survival of Aldrichina grahami (Aldrich, 1930). The results showed that 1) the developmental time to reach the pupal instar was statistically slower for the larvae reared on rabbit mince containing MA than for the control; 2) the mean length of the larvae exposed to MA concentrations was longer than those of the control; 3) the mean weight of the pupae exposed to the highest concentration of MA was significantly lighter than those of the control; 4) the GC-MS method can detect the content of MA in A. grahami immatures and empty puparia (EP).

Geographical Distribution, Biodiversity, and Species Richness of Medically Important Necrophagous Flies in Central Iran.

There have been some investigations carried out about necrophagous flies and their biodiversity indices in Iran. In the present study, the species composition of flies, their richness, and biodiversity were investigated in central Iran. In a cross-sectional study, specimens were collected monthly in three different climate areas of Qom Province. Species diversity, species richness, and species evenness were calculated to estimate the species biodiversity of flies. In total, 7,291 flies were collected and identified using valid identification keys that classified flies into four families and 15 different species. The largest collection of flies was obtained in the mountainous areas comprising 36.2% of specimens. Moreover, five forensically important species of fly were collected in this region with the potential to cause human and animal myiasis: Calliphora vicina, Lucilia sericata, Musca domestica, Wohlfahrtia nuba, and Chrysomya albiceps. There were slight differences in species richness and diversity indices in the areas. In mountainous areas, the Shannon-Wiener index and richness were maximum (H' = 1.87, S = 13). The evenness based on the Shannon evenness index in the mountainous areas was 0.728 and 0.719 in desert areas, whereas it was the highest (0.764) in the semidesert areas. Moreover, high values of the Jaccard similarity index were observed in captured necrophagous fly species in all the areas. The necrophagous fly fauna and species diversity indices were high because of an increase in the evenness. In general, some well-known forensically important species of flies were collected that proved the potential of myiasis occurrence in this area.

Necrophagous Fly Assembly: Evaluation of Species Bait Preference in Field Experiments.

The assembly of species that colonize animal organic matter, their relative abundance, and dynamics are affected by the environmental and biogeographical conditions to which these resources are exposed. Baited trap studies are essential for research on the diversity, seasonality, distribution and population dynamics of necrophagous flies. Decomposing baits provide the necessary stimulus for flies to aggregate on them. In this study, three types of bait of animal origin with different organic chemical composition were compared in terms of the diversity, richness, abundance, and species composition of saprophagous flies species that were attracted to them. Bone-meal (BM), cow liver (CL), and rotten chicken viscera (CV) were used as bait to collect flies. In total, 3,387 Calliphoridae, Sarcophagidae, and Muscidae adult flies were collected. The most abundant species were Lucilia ochricornis (Wiedemann 1830), Chrysomya albiceps (Wiedemann 1819) (Diptera: Calliphoridae), Peckia (Sarcodexia) lambens (Wiedemann 1830) (Diptera: Sarcophagidae), and Ophyra capensis (Wiedemann 1818) (Diptera: Muscidae). The type of bait had significant effects on both the total richness (F2,18 = 57.08; P < 0.0001) and the effective number of species (F2,18 = 12.81; P = 0.0003) per trap. The average richness was higher in traps baited with chicken viscera, followed by cow liver and finally by bone-meal. The composition of cow liver and bone meal species constitute subsets of the species collected with chicken viscera, thus using the three baits would not increase the number of species detected. These results indicate that chicken viscera is the most efficient bait for testing or assessing necrophagous fly diversity.

Detection of fly artifacts from four species of necrophagous flies on household materials using immunoassays.

An immunoassay was previously developed as a technique to improve methods for detection and analysis of fly artifacts found at crime scenes. The dot blot assay utilized a polyclonal antiserum (anti-md3) based on a unique digestive cathepsin D found in cyclorrhaphous Diptera. In this study, artifacts produced by adults of Calliphora vicina, Cynomya cadaverina, Sarcophaga bullata, and Protophormia terraenovae were examined using the immunoassay to determine if insect-derived stains could be distinguished from a range of human body fluid stains. A lift technique was developed which permitted transfer of fly artifacts from test materials to filter paper for dot blot analyses. All species readily deposited artifacts on all test household materials regardless of diet consumed. Despite differences in texture and porosity of the household materials, artifacts of all species transferred to the filter paper. With all fly species, anti-md3 serum bound to artifacts produced after feeding on semen, blood, feces, urine, and saliva. By contrast, anti-md3 serum did not react with any of the human fluids tested, nor with any of the lifts from household materials not exposed to flies. There was no evidence of false positives with any of the fly species tested, regardless of diet consumed. There was also no indication of false negatives with any of the dot blot assays. These observations suggest that immunoassays using anti-md3 serum performed on a simple lift of suspected fly artifacts can be used effectively as a confirmatory assay to distinguish fly regurgitate and fecal stains from human body fluids.

Molecular Identification of Necrophagous Dermestes Species in South Korea Using Cytochrome c Oxidase Subunit I Nucleotide Sequences (Genus Dermestes).

Species identification of necrophagous insects found on a dead body is an essential key in applying medicolegal entomology to the estimation of postmortem interval (PMI). Due to limited morphological identification of insect evidence, several studies have identified species using molecular information such as DNA markers. While considerable cytochrome c oxidase subunit I (COI) gene sequence data of necrophagous fly species have been collected and annotated, those of necrophagous beetle species have not. Since necrophagous beetles such as Dermestes species have a larval period longer than that of flies, beetles are useful in even the late decomposition phase in estimating minimum PMI. To obtain the full-length COI gene sequences of six Dermestes species collected from South Korea, we designed primers for polymerase chain reaction amplification and sequencing. The obtained full COI nucleotide sequences were used for performing phylogenic analysis and comparison with previously reported sequences. The results demonstrated that the COI gene sequences could be used to identify forensically important Dermestes species in South Korea.

Successive waves of dipteran flies attracted to warfarin-intoxicated rabbit carcasses in Cairo, Egypt

BackgroundForensic entomology is an increasing area of research, focuses on the violent crime, and deals with the necrophagous-feeding insects that typically decompose carcasses. The present study aimed to update the baseline data of the decay process and its associated successive waves of necrophagous flies on rabbits placed in an urban city (Cairo, Egypt) in spring season.ResultsSix rabbits divided equally into two groups were used. The first group was killed by asphyxia via hanging and marked as the control (C), while the second group was intoxicated with rodenticide warfarin (WI). The fresh, bloated, active decay, advanced decay, and skeletal stages of decomposition were observed and defined in all carcasses. Out of 30 insect families collected, 3 families were more common (Calliphoridae, Muscidae, and Sarcophagidae). Thirteen insect families were collected of which Calliphoridae, Muscidae, and Sarcophagidae were common. A total of 10,121 individuals (immature and adult stages) of 26 species were collected, of which Chrysomya rufifacies was the most common species on both types of carcasses (14.7%) followed by Musca domestica, Chrysomya megacephala, Chrysomya albiceps, Muscina stabulans, Sarcophaga hertipes, Calliphora vicina, Musca sorbens, Lucilia cuprina, Sarcophaga argyrostoma, and Lucilia sericata (2.0–12.4%). The other 15 species were represented by small numbers (cumulatively 2.9%). Among the collected flies, Scathophaga litorea (Scathophagidae) was reported for the first time as a new report in Egypt. Unexpectedly, significantly more adults and immatures were attracted to WI than to C carcasses (P < 0.05).ConclusionThis study investigated the rabbit carrion dipteran necrophagous fauna as a model which may be of value for medico-legal cases.

Impact of Constant Versus Fluctuating Temperatures on the Development and Life History Parameters of Aldrichina grahami (Diptera: Calliphoridae).

Necrophagous fly species are commonly used as forensic tools to estimate the minimum postmortem interval (PMImin). Many researchers raised necrophagous flies under constant temperature regimes to collect their developmental data. However, in most forensic cases, the ambient temperature fluctuates. In order to investigate a forensically important fly developmental mode (the Isomegalen diagram, Isomorphen diagram and Thermal summation models) and make comparisons of the developmental rate between constant temperatures and fluctuating temperatures, we used Aldrichina grahami (Diptera: Calliphoridae) to investigate the life history parameters at eight constant temperatures ranging from 8 to 36 °C. We also compared developmental rate of A. grahami in three groups of constant versus fluctuating temperatures: 8 °C vs. 6–12 °C, 12 °C vs. 10–16 °C, and 16 °C vs. 14–20 °C. Our data showed that A. grahami is cold tolerant with a mean (±SE) development threshold temperature (D0) of 3.41 ± 0.48 °C and a thermal summation constant (K) of 8125.2 ± 288.4-degree hours. The three groups subjected to fluctuating temperatures took longer to develop compared to those developing in constant temperatures when simulated in a model. These results not only provide detailed developmental data for the use of A. grahami in the estimation of the PMI, but also indicate that ambient temperature fluctuation must be taken into consideration for the use of all necrophagous fly species.