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Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae), is a blowfly species widely studied in medical, veterinary, and entomological research. Our study examined the impact of constant (15, 20, 25, 30, and 35 °C) and variable (ranging from 21.0 to 25.4 °C, with an average of 23.31 °C) temperatures on the development and larval body length of C. megacephala. Additionally, we analyzed the age of the adult C. megacephala through pteridine content and related metabolic genes analysis. Our findings revealed three distinct growth patterns: isomorphen diagram, isomegalen diagram, and thermal accumulated models. At constant temperatures of 15, 20, 25, 30, and 35 °C, egg-hatching times were 44.5 ± 8.9, 26.7 ± 4.6, 12.6 ± 1.1, 11.0 ± 1.0, and 9.9 ± 1.9 h, respectively, while it was 15.3 ± 5.9 h at variable temperatures. The total development times from oviposition to adult eclosion in C. megacephala required 858.1 ± 69.2, 362.3 ± 5.9, 289.6 ± 17.8, 207.3 ± 9.3, and 184.7 ± 12.1 h at constant temperatures of 15, 20, 25, 30, and 35 °C, respectively. This duration was extended to 282.0 ± 64.1 h under variable temperatures. However, no significant differences were found in hatching times and the total developmental durations between 25 °C and variable temperatures. A developmental threshold temperature (D 0 ) of 9.90 ± 0.77 °C and a thermal summation constant (K) of 4244.0 ± 347.0° hours were ascertained. Pteridine content patterns varied significantly across constant temperatures, but not between 25 °C and variable temperatures. Sex and temperature were identified as the primary factors influencing pteridine levels in the head of C. megacephala. Gene expression associated with pteridine metabolism decreased following adult eclosion, matching with increased pteridine concentration. Further investigations are needed to explore the use of pteridine cofactors for age-grading adult necrophagous flies. These findings provide valuable insights into the lifespan of C. megacephala, thereby offering valuable groundwork for forthcoming investigations and PMImin determination., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Heat tolerance is a distinct abiotic factor affecting the distribution and abundance of insects. Gut microbiota can contribute to host fitness, thereby increasing resistance to abiotic stress conditions. In this study, Sarcophaga peregrina is closely associated with human life in ecological habits and shows remarkable adaptability to daily and seasonal temperature fluctuations. To date, the role of gut microbiota in S. peregrina response to heat stress and its influence on the host phenotypic variability remain poorly studied., Methods: We exposed S. peregrina to heat stress at 40 °C for 3 h every day throughout the developmental stages from newly hatched larva to adult, after which gut DNA was extracted from third-instar larvae, early pupal stage, late pupal stage, and newly emerged adults, respectively. Then, 16S rRNA microbial community analyses were performed., Results: Firstly, we analyzed whether heat stress could have an impact on the life history traits of S. peregrina and showed that the growth rate of larvae was higher and the developmental time was significantly shorter after heat stress. We then proposed the role of the gut microbiota in the heat tolerance of S. peregrina, which indicated that the bacterial abundance and community structure changed significantly after heat tolerance. In particular, the relative abundance of Wohlfahrtiimonas and Ignatzschineria was higher in the third-instar larval larvae; the former increased and the latter decreased significantly after heat stress. To further explore the effect of disturbing the microbial community on thermotolerant phenotype, newly hatched larvae were fed with amikacin under heat stress, which indicated that the larval length and the whole developmental cycle was significantly shorter., Conclusion: This study indicated that Wohlfahrtiimonas and Ignatzschineria should play an important role in the post-feeding stage under heat stress, but further study is still needed. In general, heat tolerance can affect the gut microbial community structure, which in turn affects the fitness of the host., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Background: The flesh fly, Sarcophaga peregrina (Diptera: Sarcophagidae), is an important hygiene pest, that causes myiasis in humans and other mammals, typically livestock, and as a vector for various parasitic agents, including bacteria, viruses, and parasites. The role of long non-coding RNAs (lncRNAs) in regulating gene expression during metamorphosis of the flesh fly has not been well established., Methodology/principal Findings: In this study, we performed genome-wide identification and characterization of lncRNAs from the early pupal stage (1-days pupae), mid-term pupal stage (5-days pupae), and late pupal stage (9-days pupae) of S. peregrina by RNA-seq, and a total of 6921 lncRNAs transcripts were identified. RT-qPCR and enrichment analyses revealed the differentially expressed lncRNAs (DE lncRNAs) that might be associated with insect metamorphosis development. Furthermore, functional analysis revealed that the DE lncRNA (SP_lnc5000) could potentially be involved in regulating the metamorphosis of S. peregrina. RNA interference of SP_lnc5000 caused reduced expression of metamorphosis-related genes in 20-hydroxyecdysone (20E) signaling (Br-c, Ftz-F1), cuticle tanning pathway (TH, DOPA), and chitin related pathway (Cht5). Injection of dsSP_lnc5000 in 3rd instar larvae of S. peregrina resulted in deformed pupae, stagnation of pupal-adult metamorphosis, and a decrease in development time of pupal, pupariation rates and eclosion rates. Hematoxylin-eosin staining (H&E), scanning electron microscope (SEM) observation and cuticle hydrocarbons (CHCs) analysis indicated that SP_lnc5000 had crucial roles in the metamorphosis developmental by modulating pupal cuticular development., Conclusions/significance: We established that the lncRNA SP_lnc5000 potentially regulates the metamorphosis of S. peregrina by putatively affecting the structure and composition of the pupal cuticle. This study enhances our understanding of lncRNAs as regulators of metamorphosis in S. peregrina, and provide valuable insights into the identification of potential targets for vector control and the development of effective strategies for controlling the spread of myiasis and parasitic diseases., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Shang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Sarcophaga peregrina (Robineau-Desvoidy, 1830) (Diptera: Sarcophagidae) is a forensically important flesh fly with potential value for estimating the minimum postmortem interval (PMI min ). Here, the developmental patterns of S. peregrina were investigated at 5 constant temperatures (15-35 °C). Morphological changes at different developmental stages of the pupa were observed at 4 constant temperatures (15-30 °C) by removing the puparium and staining the pupa with hematoxylin and eosin. Furthermore, differentially expressed genes (DEGs) were analyzed at 25 °C in the intrapuparial period to estimate the age of S. peregrina during the intrapuparial stage. S. peregrina completed development from larviposition to adult eclosion at 15 °C, 20 °C, 25 °C, and 30 °C; the developmental durations were 1090.3 ± 30.6 h, 566.6 ± 21.9 h, 404.6 ± 13.01 h, and 280.3 ± 4.5 h, respectively, while the development could not be completed at 35 °C. The intrapuparial period of S. peregrina was divided into 12 sub-stages on the basis of the overall external morphological changes; 6 sub-stages on the basis of individual morphological structures such as the compound eyes, antennae, thorax, legs, wings, and abdomen; and 10 sub-stages on the basis of internal morphological changes detected using histological analysis. The period of each sub-stage or structure that appeared was determined. Moreover, we found that 6 genes (NDUFS2, CPAMD8, NDUFV2, Hsp27, Hsp23, and TPP) with differential expression can be used for the precise age estimation of S. peregrina during the intrapuparial period. This study provided basic developmental data for the use of S. peregrina in PMI min estimation, and we successfully estimated PMI min in a real forensic case by using a multimethod combination., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Sarcophaga (Liosarcophaga) angarosinica (Rohdendorf, 1937) (Diptera: Sarcophagidae) is a species of both medical and ecological significance. In this study, the complete mitochondrial genome (mitogenome) of S. angarosinica was sequenced and characterized. The mitogenome has a total length of 15,215 bp, including 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and an adenine and thymine-rich region. This mitogenome comprises 39.5% adenine, 9.4% guanine, 14.4% cytosine, and 36.8% thymine. Phylogenetic analysis revealed that S. angarosinica is closely related to Sarcophaga similis. This study enriches the genetic data on S. angarosinica and will contribute to establishing the phylogenetic relationships among flesh flies. [ABSTRACT FROM AUTHOR]

The intestinal bacteria of insects are crucial to the growth and development of the host. It has been found that various physiological processes of insects, such as immune response, metabolism, reproductive ability, and growth and development, involve the gastrointestinal flora. However, many external factors affect the composition of insects' intestinal microorganisms, such as the type of dietary substrate. Sarcophaga peregrina (Robineau-Desvoidy, 1830) (Diptera: Sarcophagidae) is of great significance in medicine and forensic science. In this study, we investigated the effects of ciprofloxacin on the growth and gut microbiota of S. peregrina. The results demonstrated that the maximum body length of larvae was not affected by ciprofloxacin, while the growth rate of body length quickened as the concentration of the drug increased. The weight of the pupa and adult was reduced significantly due to the effect of ciprofloxacin. After analyzing the gut microbiota composition of S. peregrina in different drug groups, it was indicated that Ignatzschineria, Providencia, Wohlfahrtiimonas, Proteus, Myroides, and Bacteroides play important roles in the growth of S. peregrina. However, they still need to be further studied. In general, ciprofloxacin can affect the gut microbial community structure, which in turn affects the fitness of the host. [ABSTRACT FROM AUTHOR]

Simple Summary: Sarcophaga crassipalpis Macquart, 1839 (Diptera: Sarcophagidae) is a flesh fly species of medical, veterinary, and forensic importance. It is often used in the laboratory for several biological studies. In the current research, we investigate its life history under changing temperatures ranging from 15.7 to 31.1 °C, with an average of 24.55 °C, and the relative humidity ranges from 31.4 to 82.8% and at six fixed temperatures of 15, 20, 25, 30, 32, and then 35 °C. Also, pteridine from the head was used to assess adult age grading. Our results revealed that the life history and development rate of S. crassipalpis under changing temperatures were very close to those observed at a fixed temperature of 25 °C. The longest and shortest growing times were found at a low rearing temperature of 15 °C and higher temperature of 32 °C, respectively. The pattern of pteridine increase differed depending on the temperature. By employing a changing temperature model in the current study, we aim to provide information on the life history of S. crassipalpis that will be useful in future research. Flesh flies (Diptera: Sarcophagidae) are regarded as significant in medical and veterinary entomology, and their development models can be utilized as considerable markers to ascertain the minimum postmortem interval (PMImin). In this research, we explored the growth cycle and larval body length of Sarcophaga crassipalpis Macquart 1839 (Diptera: Sarcophagidae) reared under variable temperatures ranging from 15.7 to 31.1 °C, with an average of 24.55 °C and relative humidity ranges from 31.4 to 82.8% and at six fixed temperatures of 15, 20, 25, 30, 32, and then 35 °C. Moreover, pteridine from the head was used to assess adult age grading. Our results allowed us to provide three development models: the isomorphen chart, the isomegalen chart, and the thermal summation models. The time taken for S. crassipalpis to complete its development from larviposition to adult emergence at constant temperatures of 15, 20, 25, 30, 32, and 35 °C was 1256.3 ± 124.2, 698.6 ± 15.1, 481.8 ± 35.7, 366.0 ± 13.5, and 295.8 ± 20.5 h, respectively, except 35 °C, where all pupae were unable to attain adulthood. They lasted 485.8 ± 5.4 h under variable temperatures. The minimum developmental limit (D0) temperature and the thermal summation constant (K) of S. crassipalpis were 9.31 ± 0.55 °C and 7290.0 ± 388.4 degree hours, respectively. The increase in pteridine content exhibited variations across different temperatures. There was quite a considerable distinction in the pteridine contents of male and female S. crassipalpis at 15 °C (p = 0.0075) and 25 °C (p = 0.0213). At 32 °C and variable temperatures, the pteridine content between female and male S. crassipalpis was not statistically divergent. However, temperature and gender remain the main factors influencing the pteridine content in the head of S. crassipalpis. We aim to provide detailed developmental data on S. crassipalpis that can be used as a valuable resource for future research and PMI estimation. [ABSTRACT FROM AUTHOR]

Simple Summary: Constant and fluctuating temperatures can affect the growth and development of insects, which will affect the accuracy of minimum postmortem interval estimation. Our study found that the development time of S. peregrina was prolonged under fluctuating temperatures. The estimation of the pupal stage has a significant effect on the estimation of PMI. We explored the feasibility of different methods for intra-puparial age estimation and found that the DEG expression profile, ATR-FTIR, and CHCs detection, combined with chemometrics, had the potential to estimate the intra-puparial age of S. peregrina under constant and fluctuating temperatures. Sarcophaga peregrina (Robineau-Desvoidy, 1830) has the potential to estimate the minimum postmortem interval (PMImin). Development data and intra-puparial age estimation are significant for PMImin estimation. Previous research has focused on constant temperatures, although fluctuating temperatures are a more real scenario at a crime scene. The current study examined the growth patterns of S. peregrina under constant (25.75 °C) and fluctuating temperatures (18–36 °C; 22–30 °C). Furthermore, differentially expressed genes, attenuated total reflectance Fourier-transform infrared spectroscopy, and cuticular hydrocarbons of S. peregrina during the intra-puparial period were used to estimate age. The results indicated that S. peregrina at fluctuating temperatures took longer to develop and had a lower pupariation rate, eclosion rate, and pupal weight than the group at constant temperatures did. Moreover, we found that six DEG expression profiles and ATR-FTIR technology, CHCs detection methods, and chemometrics can potentially estimate the intra-puparial age of S. peregrina at both constant and fluctuating temperatures. The findings of the study support the use of S. peregrina for PMImin estimation and encourage the use of entomological evidence in forensic practice. [ABSTRACT FROM AUTHOR]

Simple Summary: The precise pupal age estimates of S. peregrina are significant for estimating the minimum postmortem interval (PMImin) in forensic investigations. In the present study, we first investigated the potential of attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy and cuticular hydrocarbons (CHCs) for the age estimations of S. peregrina pupae at different constant temperatures (20 °C, 25 °C, and 30 °C). The orthogonal projections to latent structure discrimination analysis (OPLS-DA) and partial least squares (PLS) models were used to distinguish the pupae samples of different developmental ages. Sarcophaga peregrina (Robineau-Desvoidy, 1830) (Diptera: Sarcophagidae) is a forensically important flesh fly that has potential value in estimating the PMImin. The precise pupal age estimation has great implications for PMImin estimation. During larval development, the age determination is straightforward by the morphological changes and variation of length and weight, however, the pupal age estimation is more difficult due to anatomical and morphological changes not being visible. Thus, it is necessary to find new techniques and methods that can be implemented by standard experiments for accurate pupal age estimation. In this study, we first investigated the potential of attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy and cuticular hydrocarbons (CHCs) for the age estimations of S. peregrina pupae at different constant temperatures (20 °C, 25 °C, and 30 °C). The orthogonal projections latent structure discrimination analysis (OPLS-DA) classification model was used to distinguish the pupae samples of different developmental ages. Then, a multivariate statistical regression model, partial least squares (PLS), was established with the spectroscopic and hydrocarbon data for pupal age estimations. We identified 37 CHCs with a carbon chain length between 11 and 35 in the pupae of S. peregrina. The results of the OPLS-DA model show a significant separation between different developmental ages of pupae (R2X > 0.928, R2Y > 0.899, Q2 > 0.863). The PLS model had a satisfactory prediction with a good fit between the actual and predicted ages of the pupae (R2 > 0.927, RMSECV < 1.268). The results demonstrate that the variation tendencies of spectroscopy and hydrocarbons were time-dependent, and ATR-FTIR and CHCs may be optimal for the age estimations of pupae of forensically important flies with implications for PMImin estimation in forensic practice. [ABSTRACT FROM AUTHOR]

The weathering time of empty puparia could be important in predicting the minimum postmortem interval (PMImin). As corpse decomposition progresses to the skeletal stage, empty puparia often remain the sole evidence of fly activity at the scene. In this study, we used empty puparia of Sarcophaga peregrina (Diptera: Sarcophagidae) collected at ten different time points between January 2019 and February 2023 as our samples. Initially, we used the scanning electron microscope (SEM) to observe the surface of the empty puparia, but it was challenging to identify significant markers to estimate weathering time. We then utilized attenuated total internal reflectance Fourier transform infrared spectroscopy (ATR-FTIR) to detect the puparia spectrogram. Absorption peaks were observed at 1064 cm−1, 1236 cm−1, 1381 cm−1, 1538 cm−1, 1636 cm−1, 2852 cm−1, 2920 cm−1. Three machine learning models were used to regress the spectral data after dimensionality reduction using principal component analysis (PCA). Among them, eXtreme Gradient Boosting regression (XGBR) showed the best performance in the wavenumber range of 1800–600 cm−1, with a mean absolute error (MAE) of 1.20. This study highlights the value of refining these techniques for forensic applications involving entomological specimens and underscores the considerable potential of combining FTIR and machine learning in forensic practice. • Morphology observed by SEM is helpless in estimating puparium weathering time. • Most absorption peaks in the waveband region of 1800–600 cm−1. • XGBR perform well with the spectral data to estimate weathering time. [ABSTRACT FROM AUTHOR]

As our study showed, the PLS, SVR, and ANN models based on I S. peregrina i puparial hydrocarbons could be used to estimate the weathering time of the empty puparium. In this study, the results indicated that the hydrocarbon of I S. peregrina i puparium changed with weathering time, which is consistent with that of Zhu (2007), who found that puparial hydrocarbons of I C. megacephala i could be used as an indicator of PMI [[11]]. Keywords: Sarcophaga peregrina; hydrocarbon; PMI; ANN EN Sarcophaga peregrina hydrocarbon PMI ANN N.PAG N.PAG 10 09/27/22 20220901 NES 220901 1. [Extracted from the article]

Simple Summary: Forensic entomology plays a major role in postmortem interval (PMI) estimation, mainly based on morphological characteristics of larvae colonized on decomposed corpses. However, weight and length cannot be used as relevant parameters to calculate the age of wandering at the larval and pupal stages. In order to identify a specific indicator that may serve as a potential target, studies on the mechanism during postembryonic development may provide a foundation to establish a molecular age determination method. Sarcophaga peregrina (Robineau-Desvoidy, 1830) is a species of medical and forensic importance. In this study, transcriptome analysis was performed to investigate gene expression profiles from the larval to pupal stages of S. peregrina, which were used to identify differentially expressed genetic markers as candidates for molecular age estimation. This study explores molecular mechanisms during postembryonic development of S. peregrina. The developmental, stage-specific gene profiles suggest genetic markers for age prediction of forensically important flies. Sarcophaga peregrina (Robineau-Desvoidy, 1830) is a species of medical and forensic importance. In order to investigate the molecular mechanism during postembryonic development and identify specific genes that may serve as potential targets, transcriptome analysis was used to investigate its gene expression dynamics from the larval to pupal stages, based on our previous de novo-assembled genome of S. peregrina. Totals of 2457, 3656, 3764, and 2554 differentially expressed genes were identified. The specific genes encoding the structural constituent of cuticle were significantly differentially expressed, suggesting that degradation and synthesis of cuticle-related proteins might actively occur during metamorphosis. Molting (20-hydroxyecdysone, 20E) and juvenile (JH) hormone pathways were significantly enriched, and gene expression levels changed in a dynamic pattern during the developmental stages. In addition, the genes in the oxidative phosphorylation pathway were significantly expressed at a high level during the larval stage, and down-regulated from the wandering to pupal stages. Weighted gene co-expression correlation network analysis (WGCNA) further demonstrated the potential regulation mechanism of tyrosine metabolism in the process of puparium tanning. Moreover, 10 consistently up-regulated genes were further validated by qRT-PCR. The utility of the models was then examined in a blind study, indicating the ability to predict larval development. The developmental, stage-specific gene profiles suggest novel molecular markers for age prediction of forensically important flies. [ABSTRACT FROM AUTHOR]

The Calliphoridae (blowflies) are significant for forensic science, veterinary management, medical science, and economic issues. However, the phylogenetic relationships within this family are poorly understood and controversial, and the status of the Calliphoridae has been a crucial problem for understanding the evolutionary relationships of the Oestroidea these years. In the present study, seven mitochondrial genomes (mitogenomes), including six calliphorid species and one Polleniidae species, were sequenced and annotated. Then a comparative mitochondrial genomic analysis among the Calliphoridae is presented. Additionally, the phylogenetic relationship of the Calliphoridae within the larger context of the other Oestroidea was reconstructed based on the mitogenomic datasets using maximum likelihood (ML) and Bayesian methods (BI). The results suggest that the gene arrangement, codon usage, and base composition are conserved within the calliphorid species. The phylogenetic analysis based on the mitogenomic dataset recovered the Calliphoridae as monophyletic and inferred the following topology within Oestroidea: (Oestridae (Sarcophagidae (Calliphoridae + (Polleniidae + (Mesembrinellidae + Tachinidae))))). Although the number of exemplar species is limited, further studies are required. Within the Calliphoridae, the Chrysomyinae were recovered as sister taxon to Luciliinae + Calliphorinae. Our analyses indicated that mitogenomic data have the potential for illuminating the phylogenetic relationships in the Oestroidea as well as for the classification of the Calliphoridae. [ABSTRACT FROM AUTHOR]

The subfamily Sarcophaginae is extremely diverse in morphology, habit and geographical distribution, and usually considered to be of significant ecological, medical, and forensic significance. In the present study, 18 mitochondrial genomes (mitogenomes) of sarcophagid flies were first obtained. The rearrangement and orientation of genes were identical with that of ancestral insects. The degrees of compositional heterogeneity in the datasets were extremely low. Furthermore, 13 protein-coding genes were evolving under purifying selection. The phylogenic relationship of the genus-group taxa Boettcheria + (Sarcophaga + (Peckia + (Ravinia + Oxysarcodexia))) was strongly supported. Four subgenera were recovered as monophyletic (Liopygia , Liosarcophaga , Pierretia , Heteronychia) in addition to Parasarcophaga as polyphyletic. The sister-relationships between S. dux and S. aegyptiaca , S. pingi and S. kawayuensis were recovered, respectively. Moreover, the molecular phylogenetic relationships among the subgenera Helicophagella , Kozlovea , Kramerea , Pandelleisca , Phallocheira , Pseudothyrsocnema , Sinonipponia and Seniorwhitea were rarely put forward prior to this study. This study provides insight into the population genetics, molecular biology, and phylogeny for the subfamily Sarcophaginae, especially for the subgeneric classification of Sarcophaga. However, compared with the enormous species diversity of flesh flies, the available mitogenomes are still limited for recovering the phylogeny of Sarcophaginae. [ABSTRACT FROM AUTHOR]

Sarcophaga tsinanensis (Fan, 1964) (Diptera: Sarcophagidae), a species from subgenus Heteronychia. In this study, we report the complete mitochondrial genome (mitogenome) of S. tsinanensis. The length of this mitogenome was 14,972 bp in total, containing 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and a non-coding control region. The arrangement of genes was the same as that of ancestral metazoan. Sarcophaga tsinanensis has a high nucleotide bias in A/T accounting for 76.80% of total nucleotides (A 39.9%, G 9.2%, C 13.9%, and T 36.9%). The result of phylogenetic analysis revealed that S. tsinanensis cluster together with species from the same subgenus Heteronychia, showing a clear monophyletic relationship. Sarcophaga tsinanensis is closely related to its sister species Sarcophaga depressifrons, Sarcophaga plotnikovi, and Sarcophaga shnitnikovi. This study provides the mitochondrial data of S. tsinanensis for further research on evolutionary relationships and species identification of flesh flies. [ABSTRACT FROM AUTHOR]

Sarcophaga caerulescens (Zetterstedt 1838) (Diptera: Sarcophagidae) belongs to Sarcophagidae, which is closely associated with human life in ecological habits and has a clear environmental preference. Sarcophaga caerulescens can be better correlated with migration and postmortem interval (PMI) inference in forensic practice. In this study, we reported the complete mitochondrial genome (mitogenome) of S. caerulescens. The length of this mitogenome was 15,720 bp in total (GenBank accession No. MW551788), containing 13 protein-coding genes (PCGs), 2 ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and a non-coding control region. Its nucleotide composition was A (39.7%), C (14.1%), G (9.4%), and T (36.9%). The phylogenetic relationships indicated that the species of S. caerulescens was closely related to S. similis. This study provides the mitochondrial data of S. caerulescens for further study of mitochondrial genome and enriches our understanding of the phylogenetic relationship of sarcophagid flies. [ABSTRACT FROM AUTHOR]

Sarcophaga polystylata (Ho, 1934) remains to be of medical and epidemiological significance. Here, we report the complete mitochondrial genome (mitogenome) of S. polystylata (GenBank accession no. MW592361). The length of this mitogenome was 15,233 bp with 39.4% A, 9.5% G, 14.3% C, and 36.8% T. The mitogenome of S. polystylata was composed of 13 protein-coding genes (PCGs), two ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and a non-coding control region. Phylogenetic tree indicated that S. polystylata and Sarcophaga peregrina clustered together closely, both of which separated clearly from other species. This study provides the mitochondrial genetic data of S. polystylata for further understanding the phylogenetic relationship of sarcophagids species. [ABSTRACT FROM AUTHOR]

Sarcophaga gracilior Chen, 1975 (Diptera: Sarcophagidae) plays a significant role in epidemiology and medicine. In this study, we first report the complete mitochondrial genome (mitogenome) of S. gracilior. This mitogenome was 15,534 bp in length (GenBank No. MW531675), comprising 13 protein-coding genes (PCGs), two ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and a non-coding control region. The arrangement of genes was identical to that of ancestral metazoan. Nucleotide composition revealed a strong A + T bias, accounting for 76.7% (A 39.6%, G 9.3%, C 14.0%, T 37.1%). The phylogenetic relationships indicated that the species of S. gracilior emerged as sister to Sarcophaga melanura. This study provides important mitochondrial data for further studying evolutionary relationships and species identification of flesh flies. [ABSTRACT FROM AUTHOR]

Hydrotaea dentipes (Diptera: Muscidae) is one of the forensically important fly species and distributed worldwide. We describe the complete mitochondrial genome of the H. dentipes for the first time. It is a circular molecule of 16,070 bp in size, consisting of A (40.10%), G (9.20%), T (38.50%), and C (12.30%). The order and structure of the genes are similar to other Muscidae species. The mitochondrial genome (mitogenome) is composed of 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNA), two ribosomal RNA genes (rRNA), and a non-coding AT-rich region. Most PCGs used the canonical putative codon to start and stop. [ABSTRACT FROM AUTHOR]

Sarcophaga dux (Diptera: Sarcophagidae) is a necrophagous flesh fly species with potential forensic value for estimating minimum postmortem interval (PMI min). The basic developmental data and precise age estimates of the pupae are significant for PMI min estimation in forensic investigations. In the present study, we investigated the development data of that species at seven constant temperatures varying from 16 °C to 34 °C, including body length changes of the larve, developmental duration and accumulated degree hours of the preadults. Several reference genes for relative quantification of the differentially expressed genes (DEGs) were firstly selected and evaluated in the pupae of different ages under different temperatures. The DEGs of the insects during the pupal period at different constant temperatures (34, 25 and 16 °C) were further analyzed for more precise age estimation. The results showed that the developmental durations of the preadults at 16, 19, 22, 25, 28, 31 and 34 °C were 1478.6 ± 18.3 h, 726.1 ± 15.8 h, 538.5 ± 0.9 h, 394.1 ± 9.5 h, 375.6 ± 10.8 h, 284.1 ± 7.3 h, and 252.5 ± 6.1 h, respectively. The developmental threshold temperature the flies was 12.27 ± 0.35 °C, and the thermal summation constant was 5341.71 ± 249.29° hours. The most reliable reference genes during the pupal period at different temperatures were found: GST1 and 18S rRNA for the 34 °C group, GST1 and RPL49 for 25 °C, and 18S rRNA and 28S rRNA for 16 °C. The four differential expression genes (Hsp60 , A-alpha , ARP , and RPL8) have the potential to be used for more precise age estimation of pupal S. dux. This work provides important basic developmental data and a more precise age estimation method for pupal S. dux , and improves the value of this species for PMI min estimation in forensic investigations. Image 1 • The basic developmental data of S. dux during different temperatures were collected. • Four appropriate reference genes at intrapuparial stage of S. dux were selected. • The expression patterns of the same DEGs at the three temperature regimes (34°C, 25°C and 16°C) were similar. [ABSTRACT FROM AUTHOR]