Explore the vast range of titles available.

Blow flies (Diptera: Calliphoridae) are frequently used in forensic investigations due to their rapid colonization of cadavers. As with other insects, environmental temperature strongly influences their developmental rates. While published research has typically explored not only the impact of the environmental temperature, but also of other factors like tissue type and drug presence on developmental rates, the influence of photoperiod on the developmental rates of forensically relevant blow fly species has remained largely underexplored. Understanding the relationship between photoperiod and developmental times is crucial, as neglecting this aspect could compromise the accuracy of minimum post-mortem interval (minPMI) estimations. The present study investigates the impact of three photoperiod conditions (0:24, 8:16, and 12:12 light:darkness) on the developmental rates of Calliphora vicina, focusing on the duration of the different immature stages and on the total developmental time. Our results revealed significant variation in the intra-puparial stage and total development time across different photoperiods. Notably, a 12:12 photoperiod led to a significantly prolonged intra-puparial stage and total development time compared to the 0:24 photoperiod, suggesting that Calliphora vicina develops faster in total darkness. These findings highlight the importance of considering photoperiod in both laboratory rearing protocols and forensic casework to improve the accuracy and reliability of minPMI estimations. In this regard, preliminary guidelines and recommendations are provided. •The photoperiod regime significantly influences the duration of blow fly intra-puparial period.•Consequently, the total developmental time is significantly affected by photoperiod.•Preliminary recommendations on photoperiod for developmental studies are provided.

Zoonotic cutaneous leishmaniasis (ZCL), caused by Leishmania major , is a neglected tropical disease affecting impoverished populations. Current treatments are limited by cost, resistance, and side effects, highlighting the need for affordable, sustainable interventions. Lucilia sericata larvae, used in maggot therapy, effectively treat chronic wounds through debridement, antimicrobial activity, and healing promotion. This study explores how L. sericata processes L. major and proposes its potential application in ZCL treatment. The life cycles of L. sericata and L. major were maintained in laboratory conditions. Larval-parasite interactions were tested across substrates [hen liver, rat spleen, Roswell Park Memorial Institute (RPMI) 1640 cell culture medium] and time intervals (30–240 minutes). Extracorporeal effects were evaluated using trypan blue exclusion and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays; intracorporeal interactions via microscopy and nested-PCR targeting L. major rRNA genes. L. sericata excretion/secretion products and microbiota exhibited strong anti-leishmanial activity. Promastigotes were deformed within 1 hour post-exposure (hpe), fully inactivated at 4 hpe, and lysed by 6 hpe. In RPMI medium, the treatment group ( L. sericata  +  L. major ) showed significant reductions in active parasites and viable cells compared to controls after 4 hours. Microscopy revealed no parasites in larval guts, but PCR detected L. major DNA in all specimens, suggesting partial digestion. This study demonstrates that L. sericata can eliminate L. major through intra- and extra-oral digestion, supporting its potential as a biotherapeutic agent for ZCL-associated wounds. These findings offer a foundation for developing larval therapy protocols in dermatology. Further studies in animal models and clinical trials are required to validate this approach for managing ZCL.

Flies, especially those from the Calliphoridae family, play a crucial role in decomposition and are the first to colonize a cadaver. Firstly, accurate species identification is a prerequisite for entomological evidence-based calculation of postmortem interval (PMI). While morphological criteria for identifying the species of adult blow flies exist, there are either absent or inadequate keys for younger stages. In all phases of blow fly development, molecular identification offers a quick and accurate procedure. It is widely known that mitochondrial cytochrome oxidase subunit I has the capacity for molecular identification but is ineffective in certain species. This study was conducted to assess the effectiveness of the cytochrome oxidase 1 gene in the identification of seventeen different species of calliphorid flies involving four genera, Calliphora , Chrysomya , Lucilia , and Hemipyrellia . In West Bengal, 2,977 blow fly specimens were gathered from four distinct geo-climatic zones. COI barcodes were able to confirm morphological identification through low K2P intraspecific genetic divergences (0% to 1%) and moderate to high K2P interspecific genetic divergences (0.39% to 12.29%). The Neighbour-Joining (NJ) analysis demonstrated well-supported reciprocal monophyly among the species. The species grouping was in agreement with morphological and molecular identifications. The four delimitation methods, BIN, ASAP, PTP, and GMYC, used for species identification produced similar results and facilitated the proper identification of species. Therefore, it can be concluded that COI barcodes are a highly successful alternative for the molecular identification of blow flies, facilitating forensic cases and biodiversity research in India.

Myiasis is a parasitic infestation of soft vertebrate tissues by larval stages of Diptera. We briefly described the lesion-causing genus Cordylobia Grünberg (Diptera: Calliphoridae). Three Polish travelers to Uganda, Gambia, and Senegal returned with furuncular myiasis. To identify the third-instar larvae removed from their skin, we examined the morphological features of the 3 specimens and sequenced a 5’ barcoding fragment of the cytochrome c oxidase subunit I gene (COI-5P). One larva was identified as C. rodhaini Gedoelst, and 2 larvae were identified as C. anthropophaga (Blanchard). We were the first to submit the COI-5P of C. rodhaini to GenBank and the Barcode of Life Database. This is the first record of the importation of C. anthropophaga and the second record of the importation of C. rodhaini to Poland.

Chrysomya rufifacies (Diptera: Calliphoridae) is a blow fly species of forensic importance, documented to have a strong preference for colonisation of substrate already inhabited by heterospecific blow fly larvae, thus exhibiting secondary colonisation behaviour. Larvae exhibit predatory behaviour that may be useful to support development where food substrate is limited or high competition exists, but they may alternately be drawn to pre-colonised substrate to capitalise on the advantages of collective exodigestion by previous/current colonisers. Previous authors have suggested female Ch. rufifacies may use visual orientation to detect substrate currently colonised by heterospecific larvae, rather than chemoreception of volatile organic compounds (VOCs), that signify condition of substrate, which would infer that active colonisation is likely a more important oviposition cue for Ch. rufifacies than substrate condition. This study addressed attraction as well as oviposition, examining whether the condition of substrate (either previously colonised or never colonised) or the presence of heterospecific larvae was more important in the initial choice of food source by female Ch. rufifacies where conspecifics were not present, and whether the condition of substrate and presence of heterospecific larvae affects the number of offspring deposited by a female. Attraction was studied using a Y-olfactometer system, and oviposition using a binary-choice assay, with females responding to pairwise choice between an array of meat conditions (fresh, larval aged or aged) and presence/absence of Lucilia sericata larvae. Females displayed a hierarchy of choice of larval aged substrate > aged substrate > fresh substrate, with the active presence of heterospecific larvae a secondary factor in choice. Females produced higher offspring numbers on meat that was either currently or previously colonised by heterospecific larvae, demonstrating the importance of heterospecific indicators of previous or current colonisation as an oviposition cue. This serves as an important consideration for entomologists working with Ch. rufifacies in any capacity where other blow fly species may be present, and most importantly for forensic entomologists where time of colonisation is utilised to estimate PMI.

Phormia regina (Meigen, 1826; Diptera: Calliphoridae) is a Holarctic species that rapidly colonizes carcasses and has been used as an indicator for determining the minimum postmortem interval. However, studies using morphological methods to estimate the intrapuparial age of P. regina are lacking. In this study, morphological changes within the puparium were observed under a stereomicroscope at 7 constant temperatures ranging from 16 °C to 34 °C.The intrapuparial period was categorized into 12 substages. Morphological indicators, including compound eyes, mouthparts, antennae, thorax, legs, wings, and abdomen, were recorded in detail. The observed morphological changes were divided into 6–10 substages, and the duration of each substage was also recorded in detail.The results of this study provide primary data for using the intrapuparial morphology of P. regina when pupae are collected at a crime scene and estimating the minimum postmortem interval.

Addressing global food waste requires innovative and sustainable solutions. This study investigates the potential of Chrysomya megacephala (Diptera: Calliphoridae) larvae to convert food waste into valuable byproducts, while evaluating the antimicrobial properties of larval extracts. Under controlled laboratory conditions, the larvae reduced the weight of food waste by 21.87%, demonstrating their effectiveness in waste degradation. The optimal food waste-to-sawdust ratio was identified as 10:1. The bioconversion process resulted in 111.60-fold increase in larval biomass when reared on food waste and a 153.20-fold increase on fresh pork liver, highlighting their efficiency in converting protein-rich substrates. Larval extracts demonstrated significant antimicrobial activity against Bacillus subtilis and Pseudomonas aeruginosa , with minimum inhibitory concentrations (MICs) of 100 µg/ml. Proteomic analysis revealed proteins with potential antimicrobial and antioxidative properties. Furthermore, the extracts promoted cell growth in vitro without showing cytotoxic effects on HaCaT cell lines, suggesting potential applications in wound healing and infection control. These findings highlight the capacity of C. megacephala larvae to reduce food waste while generating antimicrobial agents, offering a sustainable approach to waste management with promising implications in medical applications.

The minimum postmortem interval (PMImin) could be evaluated from the developmental stage of forensically important insects colonize a corpse, such as blow flies (Diptera: Calliphoridae). Unlike larvae, the developmental stage of which is well established according to their morphology, estimating the age of pupae is proven to be challenging. Recently, several studies reported the regulation of special genes during the development of blow fly pupae. However, gene regulation in Aldrichina grahami during the intrapuparial period remains to be studied. Therefore, we set out to investigate the mRNA levels of heat shock protein 23 (Hsp23), heat shock protein 24 (Hsp24), and 1_16 during the metamorphosis of A. grahami pupae. First, we examined seven candidate reference genes (ribosomal protein 49 (RP49), 18S ribosomal RNA (18S rRNA), 28S ribosomal RNA (28S rRNA), beta-tubulin at 56D (β-tubulin), Ribosomal protein L23 (RPL23), glutathione S-transferase (GST1), and Actin. Three widely used algorithms (NormFinder, BestKeeper, and geNorm) were applied to evaluate the mRNA levels of reference gene candidates in puparium at three stable temperatures (15, 22, and 27°C). Next, mRNA expression of Hsp23, Hsp24, and 1_16 during A. grahami metamorphosis was examined. We demonstrated that mRNA expression levels of Hsp23, Hsp24, and 1_16 showed time-specific regulation. In summary, our study identified three gene markers for the intrapuparial period of A. grahami and might provide a potential application in PMImin estimation.

In the medico-legal application of forensic entomology, estimating the time of death is critical and traditionally relies on changes in observable traits of carrion feeding insect larvae. Traits such as size, weight, and morphology can be used to predict the insect specimen age and help define the minimum time since death. The blowfly Phormia regina Meigen (Diptera: Calliphoridae) is a key forensic insect, yet age estimation for older maggots in this and other carrion-feeding species is particularly challenging due to the limited morphological changes in the late-stage larvae. To enhance age-estimation precision, we employed transcriptomic profiling on blowfly maggots, aiming to identify genes as markers for time of death estimation. Our study characterized maggot development, reinforcing that weight and behavior cannot precisely determine age between 100 and 130 hours at 27.5 °C. We built a chromosomal scale annotated genome, establishing a reliable database for uncovering transcriptomic signatures during larval development. Applying differential gene expression analyses, weighted gene co-expression network analysis, and the generalized linear model, we identified nine candidate genes (y5078, y5076, agt2, ech1, dhb4, asm, gabd, acohc, ivd) that delineate the age of otherwise indeterminate maggots. This research introduces a molecular approach to address a longstanding problem in forensic entomology and promises to increase precision in determining the time of death at a crime scene.

The oriental latrine fly, Chrysomya megacephala (Diptera: Calliphoridae), is a medically important synanthropic blow fly species characterized by its necrophagy and coprophagy, often observed near carrion and animal feces. Notably, C. megacephala always arrives at carcass earlier than other species. To elucidate the underlying mechanisms behind the host choice in C. megacephala , we present the chromosome-scale genome assembly for this species. The genome size is 816.79 Mb, with a contig N50 of 1.60 Mb. The Hi-C data were anchored to six chromosomes, accounting for 99.93% of the draft assembled genome. Comparative genomic analysis revealed significant expansions in pathways of ligand-gated ion channel activity, passive transmembrane transporter activity, and protein methyltransferase activity, which may be closely associated with host localization and oviposition. After identifying 69 odor-binding proteins (OBPs) in the assembled genome, phylogenetic analysis showed that DmelOBP99b and CmegOBP99b exhibited high homology. Transcriptome analysis demonstrated that the relative expression of CmegOBP99b was consistently the highest during the metamorphosis, and RT-qPCR further confirmed the similar results. Additionally, CmegOBP99b exhibited a strong binding affinity to DMDS (dimethyl disulfide) as determined by molecular docking. To determine the protein expression level of CmegOBP99b in various body parts, we prepared recombinant CmegOBP99b protein and anti- CmegOBP99b polyclonal antibodies. Western blot analysis showed that CmegOBP99b was significantly expressed in the female’s head compared to other parts, which is consistent with RT-qPCR results. Therefore, CmegOBP99b may be the primary odor-binding protein responsible for olfactory recognition and the behavioral coordination of C. megacephala . This study not only provides valuable insights into the molecular mechanisms of oviposition localization in C. megacephala but also facilitates further research into the genetic diversity and phylogeny of the Calliphoridae family.