Abeer Mohsen Salem

Blow flies of the genus Chrysomya are among the earliest colonizers of human remains and are important forensic indicators in Africa and the Middle East. However, the population genetic structure of Old World Chrysomya species in North Africa remains poorly understood. We conducted the first amplified fragment length polymorphism (AFLP)-based population genetic analysis of Chrysomya albiceps and Chrysomya marginalis in Egypt to assess geographic and temporal structure and to evaluate their forensic utility for population assignment. Adult flies were collected from multiple localities in Egypt and across consecutive years, and AFLP profiles were generated using four primer combinations. All individuals yielded unique multilocus genotypes. In C. albiceps , most genetic variation occurred within populations (93%), with only weak but significant among-population differentiation (Fst = 0.045), no isolation by distance, and no geographically coherent clustering, indicating near-panmixia at the regional scale. In C. marginalis , spatial differentiation was negligible, whereas temporal comparison of samples from Giza revealed modest year-to-year structuring. Within-sample relatedness was consistently positive in both species, suggesting nonrandom kin associations within collection events. Although simulation-based assignment success was high, empirical reallocation performance was substantially lower, indicating that assignment accuracy depends strongly on the level of genetic differentiation and may not yet be robust for routine forensic inference. This apparent paradox reflects fine-scale structuring driven by kin aggregation within sampling events, despite overall high gene flow at the regional scale.

West Nile virus (WNV) causes West Nile fever and is primarily transmitted by Culex spp. mosquitoes. The primary hosts of WNV are birds, while humans and horses can also develop illness following infection. Currently, no specific antiviral therapy exists for WNV, and vaccination remains the only effective preventive measure. This study aimed to evaluate the efficiency of the crude bee venom (BV) and its nanoemulsion (NE) on WNV by determining in vitro Vero cells cytotoxicity and viral titer by RT-qPCR. We also conducted BV peptides-envelope glycoprotein of WNV interactions by molecular docking analysis. The NE of BV was characterized by dynamic light scattering (DLS) and zeta potential analyses. The DLS revealed that the size of NE was greater than that of BV. Also, the DLS showed that the polydispersity index of NE was less than that of BV (0.349 and 0.557, respectively). Zeta potential analysis of NE was slightly less than that of BV (23.0 and 23.7 mV, respectively). Both treatments showed low cytopathic effects on Vero cells compared to positive control infected with WNV. RT-qPCR revealed that the titer of WNV was lower in cells treated with BV and its NE than in controls. Molecular docking revealed BV peptides-envelope glycoprotein of WNV interactions, with binding energy of - 5.2, - 4.5, - 13.0, - 10.8, and - 5.1 kcal/mol for the peptides apamin, tertiapin, melittin, mast cell degranulating peptide, and secapin-2 peptide, respectively. BV-NE demonstrated preliminary in vitro anti-WNV activity requiring mechanistic, toxicological, and in vivo validation.

Population genetic patterns and changes in allele frequency within blow flies can provide valuable insight into population structure, diversification, dispersal, gene flow, and population assignments. The population genetic structure of the oriental latrine blow fly, Chrysomya megacephala (Fabricius), was investigated using amplified fragment length polymorphisms (AFLP). The goal was to validate the use of this technique to create genetic profile data and determine it is feasibility for inferring postmortem relocation of corpses. The AFLP technique generated 590 polymorphic loci for C. megacephala. Analysis of molecular variation (AMOVA) found significantly high genetic variation within individuals of all fly populations, with little variation among populations from different geographic locations. STRUCTURE and principal coordinate analyses (PCoA) revealed no population structure based on geography, with weak correlation between genetic and geographic distances, and moderate temporal differentiation was noted among C. megacephala samples. Across the entire data set, the mean relative relatedness coefficients were positive, suggesting that flies arriving at the same bait (carcass) share nonrandom proportions of alleles and comprise of closely related individuals. Genetic assignment of C. megacephala flies to a putative source population resulted in a 90.81% success rate, indicating the possibility of using these flies to connect sites between which a corpse had been moved even in the absence of overall geographic population structure.

Entomological warfare has been known since ancient civilizations used insects to spread disease among enemies. Advances in science and technology have enhanced the potential for using insects as vectors for viruses and toxins in warfare, including both traditional methods and new innovations like genetic engineering. This review aims to: (i) explore the historical and contemporary roles of insects in biological warfare; (ii) examine the ethical, legal, and ecological challenges associated with this form of warfare; and (iii) outline historical uses of insects as weapons, from releasing disease-carrying mosquitoes to creating genetically modified insects. It highlights the complexities of legal and ethical frameworks governing entomological warfare and potential environmental impacts. While insects offer unique advantages as biological agents, their use in combat raises significant ethical and environmental concerns. Therefore, there is an urgent need for international cooperation and regulatory oversight to ensure responsible and ethical applications of entomological warfare technologies.

Background: Blow flies play a crucial role in determining the minimum postmortem interval (PMI_min) due to their prolific breeding on carrion. This study focuses on three important medical and forensically species investigating how fluctuating temperatures affect their biology and confirming their identification through mitochondrial DNA analysis. Results: The different biological aspects of the three studied species Lucilia sericata, Chrysomya albiceps, and Chrysomya megacephala were temperature dependent. The highest number of emerging adults, number of eggs laid per female and egg hatchability were in summer, followed by spring, autumn and winter. Preoviposition period, incubation period, maggot duration and pupal duration were inversely proportional with temperature. Adult females lived longer than males during the four seasons. The longest longevity of male and female of L. sericata was in spring. Male and female C. megacephala and C. albiceps had the longest life span in winter. Phylogenetic analysis revealed 97, 99 and 99% similarity between L. sericata, C. megacephala and C. albiceps compared to their reference species. Conclusions: Seasonal temperature variations significantly impact developmental rates of blow flies causing deviation in PMI_min estimation if these biological data were to be used. The findings highlight the importance of integrating temperature effects and molecular data for more accurate investigation and identification. Graphical

Previous research has shown that the effects of different drugs on the growth rate of blow flies vary by species, which can influence the determination of the minimum post-mortem interval (PMImin​). In this study, we assessed how morphine affects the development, growth, and survival rate of Chrysomya megacephala (Fabricius, 1794), a forensically important blow fly species, and whether these effects are dose dependent. We also measured morphine concentrations in insect samples (3rd -instar larvae and puparial cases) and in various post-mortem tissues of rabbits (cardiac blood, skeletal muscle, and liver). Rabbits were administered two lethal doses of morphine hydrochloride (5 and 10 mg/kg), then sacrificed, and tissue samples were collected for toxicological analysis. At a controlled temperature of 37 °C, newly hatched larvae (1–2 h old) were placed on the rabbit carcasses and randomly sampled at 12 h regular interval (12–132 h) to determine larval development time, puparial development time, larval mortality, larval length, and larval weight. Changes in these insect parameters led to an underestimation of PMImin by up to 15.92 h if the presence of morphine was not considered. Moreover, morphine concentrations in all insect samples (180 and 223 ng/g in the 3rd instar larvae; 40 and 45 ng/g in the puparial cases after treatment with 5 and 10 mg/kg of morphine hydrochloride, respectively) and rabbit tissues (1129 and 2564 ng/g in the cardiac blood; 1777 and 2823 ng/g in the liver; 825 and 1070 ng/g in the skeletal muscles after treatment with 5 and 10 mg/kg of morphine hydrochloride, respectively) were measured using high-performance liquid chromatography system equipped with a binary pump and UV detector. Morphine can affect the development of different life cycle stages of Ch. megacephala, highlighting the importance of accounting for its presence when determining PMImin ​ in such conditions. The data obtained could aid in criminal investigations. Although a concentration-dependent effect has been discussed, the use of whole animal carcasses may lead to uneven distribution of the drug, and concentrations in the areas from which larvae are feeding may differ significantly due to ante-mortem processes in the body.

Background: Coleoptera is the second-most significant insect group associated with decomposing carcasses, yet its role in the decomposition process and postmortem colonization following envenomation is poorly understood. Purpose of the Study: This study aimed to investigate the effects of the venoms from Cerastes cerastes and Naja haje on the decomposition of rabbit carcasses while evaluating the main beetle taxa attracted to these decaying remains. Methods: Three groups of rabbits, each with five individuals, were utilized. The first group was injected with the venom of Cerastes cerastes, the second with Naja haje venom, and the control group received 0.85% physiological saline before euthanasia with CO₂. Results: Four decomposition stages (fresh, bloating, decay, and dry) with durations varying based on venom type and carcass condition were observed. A total of 647 individual beetles of six species (Necrobia rufipes, Attagenus sp., Dermestes frischii, D. maculatus, Bledius sp., and Apentanodes sp.) belonging to four families (Cleridae, Dermestidae, Staphylinidae, and Tenebrionidae) were collected and identified. D. maculatus was the most abundant species. Fewer beetles were attracted to carcasses envenomed with N. haje compared to the other groups. Conclusions: Envenomation by snake venom influences the attraction and succession rate of necrophilous coleopterans to carcasses, which is important for forensic investigations.

Wastewater irrigation is often practiced in arid regions, which can increase the chance of heavy metals contaminating the agricultural system. This contamination poses risks to both the environment and human health. This research looked into how cadmium (Cd), lead (Pb), copper (Cu), and zinc (Zn) move through a food chain involving soil, plants, and arthropods. The study was conducted in El-Gabal El-Asfar, Egypt, comparing treated and untreated wastewater irrigation areas. Six soil-irrigated sites and one reference site were sampled for soil, alfalfa (Medicago sativa), two grasshopper species (Aiolopus thalassinus and Calephorus compressicornis), and a wolf spider (Hogna ferox). The samples were analyzed for their heavy metal content. Metal concentrations in all components of the wastewater irrigated system were significantly higher compared to the reference site. The wolf spider and the soil contained the highest levels of Cd, Pb, and Cu, while the greatest concentrations of Zn were found in the spider and grasshoppers. Despite limited transfer from soil to plant, trace elements biomagnified within the terrestrial food chain, specifically from grasshoppers to wolf spiders. The correlation analysis of metal levels between soils, plants, and arthropods in the present study reflects its transfer across the trophic levels. It suggests that dietary intake is the main source of metal accumulation in arthropods. The present study, therefore, quite clearly indicated the possibility of heavy metal biomagnification in terrestrial food chains of wastewater-irrigated agroecosystems. Continuous monitoring and management of such systems are advocated to avoid environmental and public health risks.

Background: Studying insect succession on carcasses is important in estimating the postmortem interval. This study aims to identify the decomposition stages of decomposing rabbit carcasses and to find out the relationship between seasonal variations and abundance of insects colonizing rabbit carcasses at El-Sharkia Governorate, Egypt. Methods: Three domestic rabbits (weighing 1300 g each) were killed by a sharp knife. The carcasses were exposed to the sun, left to decompose and inspected twice daily at 6-h intervals to collect insects. Maggots were collected and reared. Results: The rabbit carcasses underwent four decomposition stages: fresh, bloat, decay and dry stages. The identified families and their respective collected species included three dipteran families: Calliphoridae (Lucilia sericata, Chrysomya megacephala, and Chrysomya albiceps), Sarcophagidae (Sarcophaga argyrostoma) and Muscidae (Musca domestica and Synthesiomyia nudiseta), three coleopteran families. Histeridae (Saprinus semistriatus), Cleridae (Necrobia rufipes) and Dermestidae (Dermestes frischii and Attagenus gloriosus) and three hymenopteran families: Chalcididae (Brachymeria femorata), Vespidae (Vespa orientalis) and Formicidae (Monomorium sp.). Carrion fauna was dominated by dipteran and coleopteran species, with calliphorid and sarcophagid flies found to play a significant role in carrion consumption process. Conclusion: The succession pattern and decomposition rate were season dependent. The information collected may help establish the basic database for entomological forensic investigations in the future. Graphical