Potential therapeutic biomolecules of hymenopteran venom against SARS-CoV-2 from Egyptian patients.

The therapeutic potential of insect-derived bioactive molecules as anti-SARS-CoV-2 agents has shown promising results. Hymenopteran venoms, notably from Apis mellifera (honeybee) and Vespa orientalis (oriental wasp), were examined for the first time in an in vitro setting for their potential anti-COVID-19 activity. This assessment utilized an immunodiagnostic system to detect the SARS-CoV-2 nucleocapsid antigen titer reduction. Further analyses, including cytotoxicity assays, plaque reduction assays, and in silico docking-based screening, were performed to evaluate the efficacy of the most potent venom. Results indicated that bee and wasp venoms contain bioactive molecules with potential therapeutic effects against SARS-CoV-2.Nevertheless, the wasp venom exhibited superior efficacy compared to bee venom, achieving a 90% maximal (EC90) concentration effect of antigen depletion at 0.184 mg/mL, in contrast to 2.23 mg/mL for bee venom. The cytotoxicity of the wasp venom was assessed on Vero E6 cells 48 h post-treatment using the MTT assay. The CC 50 of the cell growth was 0.16617 mg/mL for Vero E6 cells. The plaque reduction assay of wasp venom revealed 50% inhibition (IC50) at a 0.208 mg/mL concentration. The viral count at 50% inhibition was 2.5 × 104 PFU/mL compared to the initial viral count of 5 × 104 PFU/mL. In silico data for the wasp venom revealed a strong attraction to binding sites on the ACE2 protein, indicating ideal interactions. This substantiates the potential of wasp venom as a promising viral inhibitor against SARS-CoV-2, suggesting its consideration as a prospective natural preventive and curative antiviral drug. In conclusion, hymenopteran venoms, particularly wasp venom, hold promise as a source of potential therapeutic biomolecules against SARS-CoV-2. More research and clinical trials are needed to evaluate these results and investigate their potential for translation into innovative antiviral therapies.

Isolation of Multidrug-Resistant Helicobacter pylori from Wild Houseflies Musca domestica with a New Perspective for the Treatment.

Background: High frequency of Helicobacter pylori infection and the unknown mode of transmission prompted us to investigate H. pylori-wild housefly relationship. H. pylori causes chronic gastritis, peptic ulcers, and stomach cancer. H. pylori persists in the gut of the experimentally infected houseflies. The existence of H. pylori strains isolated from wild houseflies, on the other hand, has never been documented. Materials and Methods: In this study, 902 wild houseflies from different sites were identified as Musca domestica, then 60 flies were screened by traditional microbiological techniques and H. pylori-specific 16S rRNA gene. The antibiotic resistance (ART) was investigated phenotypically. Wild housefly gut bacterial isolates were further evaluated genotypically to have 23S rRNA gene mutation related to clarithromycin resistance. To find efficient therapeutic alternatives, the potency of three plant extracts (garlic, ginger, and lemon) and the wasp, Vespa orientalis venom was evaluated against H. pylori. The cytotoxic effect of the crude wasp venom, the most potent extract, against Vero and Colon cancer (Caco2) cell lines was investigated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: All isolates from houseflies were positive. The isolated bacteria have variable resistance to frequently used antibiotics in all isolates. Minimum inhibitory concentration values of 15.625 mg/mL for both ginger and lemon extracts, 7.8125 mg/mL for garlic extract, and 0.0313 mg/mL for wasp venom were recorded. Wasp venom has the most potent antibacterial activity compared with the four antibiotics that are currently used in therapies against H. pylori. Conclusion: We conclude that wild houseflies can play a role in disseminating H. pylori. The housefly gut may be a suitable environment for the horizontal transfer of ART genes among its associated microbiome and H. pylori. Wasp venom proved its potential activity as a new and effective anti-H. pylori drug for both therapeutic and preventative usage.

The Antibacterial Activity of Egyptian Wasp Chitosan-Based Nanoparticles against Important Antibiotic-Resistant Pathogens.

The current work discusses the production and characterization of new biodegradable nanoparticles for biomedical applications based on insect chitosan. Chitosan has numerous features due to the presence of primary amine groups in repeating units, such as antibacterial and anticancer activities. When polyanion tripolyphosphate is added to chitosan, it creates nanoparticles with higher antibacterial activity than the original chitosan. In this study, the ionic gelation technique was used to make wasp chitosan nanoparticles (WCSNPs) in which TEM and FTIR were used to investigate the physicochemical properties of the nanoparticles. In addition, the antibacterial activities of chitosan nanoparticles against extended-spectrum beta-lactamase (ESBL)- and carbapenemase-producing Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa were evaluated. The extracted wasp chitosan exhibited high solubility in acetic acid and met all standard criteria of all characterization testes for nanoparticles; the zeta potential indicated stable WCSNPs capable of binding to cellular membrane and increasing the cellular uptake. The produced WCSNPs showed growth inhibition activity against all tested strains, and the bacterial count was lower than the initial count. The inhibition percent of WCSNPs showed that the lowest concentration of WCSNPs was found to be effective against tested strains. WCSNPs' antibacterial activity implies that they could be used as novel, highly effective antibacterial agents in a variety of biological applications requiring antibacterial characteristics.

Evolutionary profile of the family Calliphoridae, with notes on the origin of myiasis.

The family Calliphoridae is a group of heterogenous calyptrate flies with a worldwide distribution including species of ecological, veterinary, medical, and forensic importance. Notorious for their parasitic habits, the larvae of many blowflies are characterised - like some other dipteran larvae - by their ability to develop in animal flesh. When parasitism affects a living host, it is termed "myiasis". This has led the Calliphoridae to be considered as a pivotal family in its relationship with a man. Nevertheless, even after more than 50 years of research, the phylogenetic relationships among calliphorid subfamilies together with the evolutionary origin of myiasis remain unclear. In order to elucidate these problems, we constructed three phylogenetic trees by using nucleotide sequence data from cytochrome oxidase subunit one (COI), representing a mitochondrial conservative gene, and nuclear 28S subunit of ribosomal RNA gene (28S rRNA) in order to interpret the evolutionary profile of myiasis in the family Calliphoridae. The sequenced data represented species associated with ectoparasitic life-styles, either saprophagy or facultative and obligate parasitism. A total number of 50 accessions were collected for 28S rRNA, 56 for COI, and 38 for combined sequences phylogeny. Molecular Evolutionary Genetics Analysis (MEGA) software was used to align 2197 nucleotide positions of 28S rRNA and 1500 nucleotide positions of COI with a gap opening penalties and gap extension penalties equalling 20 and 0.1 respectively. The results reveal the non-monophyly of the family Calliphoridae despite the stable monophyletic status of the Chrysomyinae, Luciliinae, and Auchmeromyiinae. Also, our findings recommend ranking the Toxotarsinae as a separate family. Furthermore, comparative analysis of the phylogenetic trees shows that the habit of obligatory myiasis originated independently more than five times. This strengthens our hypothesis that the origin of eating fresh meat is a case of convergent evolution that has taken place after speciation events millions of years ago. Finally, estimating the divergence dates between lineages from molecular sequences provides a better chance of understanding their evolutionary biology.

Modeling current and future global distribution of Chrysomya bezziana under changing climate.

In the last few years, significant changes in climate have had a disparate effect on biodiversity. The influences of these changes are random and unpredictable. The resurgence of insect pests, especially of medical and veterinary importance, often corresponds with climate changes. The Old World screwworm, Chrysomya bezziana, is one of the most important myiasis-causing flies that parasitize warm-blooded animals in the Eastern Hemisphere. We used a spatial distribution modeling approach to estimate the consequences of climatic changes on the potential geographic distribution of this insect throughout the world currently and in the future. A Maxent model used occurrence data from 104 localities and 19 climatic factors to predict the suitable habitat regions throughout the world. Two representative concentration pathways 2.6 and 8.5, were used to forecast the future distribution of C. bezziana in 2050 and 2070. The Maxent model for C. bezziana provided a satisfactory result, with a high value of the Area Under Curve equal to 0.855 (±0.001). Furthermore, the True Skilled Statistics value is equal to 0.67. These values indicate the significant influence on the model of the ecology of this fly species. Jackknife test indicated that temperature variables play a significant role in C. bezziana dynamics. The resultant models indicated the areas at risk of invasion by potential serious medical/veterinary issues, especially in countries with a large livestock production.