The Analysis of Oral and Fecal Virome Detects Multiple Novel Emerging Viruses in Snakes

Wild animals are considered reservoirs for emerging and reemerging viruses, such as the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Previous studies have reported that bats and ticks harbored variable important pathogenic viruses, some of which could cause potential diseases in humans and livestock, while viruses carried by reptiles were rarely reported. Our study first conducted snakes' virome analysis to establish effective surveillance of potential transboundary emerging diseases. Consequently, Adenoviridae, Circoviridae, Retroviridae, and Parvoviridae were identified in oral samples from Protobothrops mucrosquamatus, Elaphe dione, and Gloydius angusticeps based on sequence similarity to existing viruses. Picornaviridae and Adenoviridae were also identified in fecal samples of Protobothrops mucrosquamatus. Notably, the iflavirus and foamy virus were first reported in Protobothrops mucrosquamatus, enriching the transboundary viral diversity in snakes. Furthermore, phylogenetic analysis revealed that both the novel-identified viruses showed low genetic similarity with previously reported viruses. This study provided a basis for our understanding of microbiome diversity and the surveillance and prevention of emerging and unknown viruses in snakes.

VSV-BASED COVID VACCINE FOR THE PREVENTION OF BREAKTHROUGH SARS-CoV-2 INFECTIONS

Background: More than 12 billion doses of COVID-19 vaccine administrations and over 630 million natural infections should have developed adequate levels of herd immunity over the last three years. However, there have been many new waves of coronavirus infections. The development of safe and effective vaccines to control breakthrough SARS-CoV-2 infections remain an urgent priority. We have developed a recombinant VSV vector-based vaccine to fulfill this worldwide need. Method(s): We have used a recombinant vesicular stomatitis virus (rVSV)-based prime-boost immunization strategy to develop an effective COVID-19 recall vaccine candidate. We have constructed an attenuated recombinant VSV genome carrying the full-length Spike protein gene of SARS-CoV-2. Adding the honeybee melittin signal peptide (msp) at the N-terminus enhanced the protein expression and adding the VSV G protein transmembrane domain and the cytoplasmic tail (Gtc) at the C-terminus of the Spike protein allowed efficient incorporation of the Spike protein into pseudotype VSV. Result(s): In immunized mice, rVSV with chimeric rVSV-msp-S-Gtc induced high levels of potent neutralizing antibodies (nAbs) and CD8+ T cell responses, while the full-length Spike with Gtc proved to be the superior immunogen. More importantly, rVSV-msp-S-Gtc-vaccinated animals were completely protected from subsequent SARS-CoV-2 challenges. Furthermore, rVSV-Wuhan and rVSV-Delta vaccines, and an rVSV-Trivalent (mixed rVSV-Wuhan, -Beta and -Delta) vaccine elicited potent nAbs against live SARS-CoV-2 Wuhan (USAWA1), Beta (B.1.351), Delta (B.1.617.2) and Omicron (B.1.1.529) viruses. Heterologous boosting of rVSV-Wuhan with rVSV-Delta induced strong nAb responses against Delta and Omicron viruses, with the rVSV-Trivalent vaccine consistently inducing effective nAbs against all the SARS-CoV-2 variants tested. All rVSV-msp-S-Gtc vaccines also elicited an immunodominant Spike-specific CD8+ T cell response. Conclusion(s): rVSV vaccines targeting SARS-CoV-2 variants of concern can be considered as an effective booster vaccine in the global fight against COVID-19.

Computational investigation of honeybee venom proteins as potential omicron SARS-CoV-2 inhibitors

Because of the catastrophic consequences of COVID-19 on the world population, there should be novel interventions to handle ongoing infections and daily death cases. The aim of the current study is to examine the effectiveness of HBV (Honeybee venom) proteins on spike protein RBD by in silico tools. The sequences of 5 HBV proteins were used for homology modeling by Phyre 2. The generated protein models were employed for protein-protein docking against Omicron Spike glycoprotein receptor binding domain (RBD) (PDB ID# 7T9L) through HDock and ClusPro platforms followed by prediction of binding affinity using PRODIGY web portal and PDBsum for revealing interaction details. It was found that all of the examined HBV proteins exhibited strong docking scores and binding affinity profiles toward RBD. The findings of the present study indicate the possible HBV as preventive as well as treatment options against Omicron SARS-CoV-2. © 2022, Palladin Institute of Biochemistry of the NASU. All rights reserved.

In silico screening of potent bioactive compounds from honeybee products against COVID-19 target enzymes.

After the early advent of the Coronavirus Disease 2019 (COVID-19) pandemic, myriads of FDA-approved drugs have been massively repurposed for COVID-19 treatment based on molecular docking against selected protein targets that play fundamental roles in the replication cycle of the novel coronavirus. Honeybee products are well known of their nutritional values and medicinal effects. Bee products contain bioactive compounds in the form of a collection of phenolic acids, flavonoids, and terpenes of natural origin that display wide spectrum antiviral effects. We revealed by molecular docking the profound binding affinity of 14 selected phenolics and terpenes present in honey and propolis (bees glue) against the main protease (Mpro) and RNA-dependent RNA polymerase (RdRp) enzymes of the novel SARS-CoV-2 virus (the causative agent of COVID-19) using AutoDock Vina software. Of these compounds, p-coumaric acid, ellagic acid, kaempferol, and quercetin have the strongest interaction with the SARS-CoV-2 target enzymes, and it may be considered an effective COVID-19 inhibitor.

Disinfectant dodecyl dimethyl benzyl ammonium chloride (DDBAC) disrupts gut microbiota, phospholipids, and calcium signaling in honeybees (Apis mellifera) at an environmentally relevant level.

One of the impacts of the Coronavirus disease 2019 (COVID-19) pandemic has been a profound increase in the application amounts of disinfectants. Dodecyl dimethyl benzyl ammonium chloride (DDBAC) is a widely used disinfectant, yet its hazards to non-target species remain largely unknown. We are unaware of any studies assessing DDBAC's impacts on honeybee, a pollinator species that is a useful indicator of environmental pollution essential for many forms of agricultural production. Here, we assessed the potentially negative effects of DDBAC on honeybees. After conducting a formal toxicity evaluation of DDBAC on honeybee mortality, we detected an accumulation of DDBAC in the honeybee midgut. We subsequently studied the midgut tissues of honeybees exposed to sub-lethal concentrations of DDBAC: histopathological examination revealed damage to midgut tissue upon DDBAC exposure, microbiome analysis showed a decreased abundance of beneficial midgut microbiota, lipidomics analysis revealed a significant reduction in cell membrane phospholipids with known functions in signal transduction, and a transcriptome analysis detected altered expression of genes involved in calcium signaling pathways (that variously function in calcium absorption, muscle contraction, and neurotransmission). Thus, our study establishes that DDBAC impacts honeybee midgut functions at multiple levels. Our study represents an early warning about the hazards of DDBAC and appeals for the proper stewardship of DDBAC to ensure the protection of our ecological environment.

Honeybee Products for the Treatment and/ or Recovery of COVID-19 and Other Coronavirus-Related Respiratory Tract Infections: A Rapid Systematic Review

Methods: A protocol applying Cochrane rapid review methods was registered with PROSPERO: (CRD42020193847) in July 2020. Four English databases plus preprint servers and trial registries were searched for randomised controlled trials (RCTs). The evidence was appraised using RoB 2.0 and GRADE-approach. Results: 27 results, derived from 9 RCTs, included 674 adults and 781 children. In Hospitalized patients with confirmed SARS-CoV-2 infections, propolis plus usual care compared to usual care alone, reduced the risk of shock, (RR:0.36;95%CI:0.15, 0.88), respiratory failure (RR:0.32;95%CI:0.11, 0.92), and kidney injury (RR:0.36: 95% CI:0.15, 0.88) (all low-certainty) and duration of hospital admission (MD:-3.81 days, 95%CI:-6.19, -1.44) (moderate-certainty). Compared to coffee, honey plus coffee and honey alone, reduced the severity of post-infectious cough of a least 3-weeks duration in adults (MD:-1.40, 95%CI:-1.67, -1.13) (low-certainty) and (MD:-0.40, 95%CI:-0.75, 0.05) (very low-certainty) respectively. Honey was less effective than Guaifenesin in reducing cough severity at 60-minutes in adults with non-specific acute RTIs (MD:0.20 points, 95%CI: 0.05, 0.35) (very low-certainty). Honey compared to placebo reduced the duration of cough in children (MD:-0.71 days 95% CI:-1.15, -0.28) and compared to salbutamol (MD:-0.54 days 95%CI:-0.99, -0.09) (low-certainty);and the global impact of paediatric nocturnal cough was reduced by honey plus usualcare compared to usual-care alone (SMD:-0.80, 95%CI:-1.28, -0.32) (very low-certainty) and pharmaceutical cough medicines (SMD:-0.75 95%CI:-1.13, -0.36) (low-certainty). Background: This rapid review systematically evaluated the effects of honeybee products compared to controls for the duration, severity, and recovery of acute viral respiratory tract infections (RTIs), including SARS-CoV-2, in adults and children. Conclusion: More RCTs are needed to robustly inform the role of honey and propolis for treatment of SARS-CoV-2 and related respiratory infections.

Voltammetric determination of heavy metals in honey bee venom using hanging mercury drop electrode and PLA/carbon conductive filament for 3D printer.

A new method for determination of selected heavy metals (Cd, Pb, Cu, Zn, and Ni) in honey bee venom was developed. Heavy metals are metabolized and incorporated into bee products, including honey and honey bee venom (apitoxin). Their composition reflects contamination of "bee environment", providing information about heavy metal contamination in the neighborhood of human dwellings. Moreover, assessment of bee products contamination is relevant for medicine, as they are a tool for promising therapeutic and chemoprophylactic strategies against COVID-19 (SARS-CoV-2). Owing to the complicated matrix, the developed method consists of wet mineralization with sulfuric acid, nitric acid, under increased temperature, and pressure and subsequent repeated boiling with concentrated nitric acid. Determination of the selected metals was carried out by anodic or cathodic stripping voltammetry on two types of electrodes: pen-type hanging mercury drop electrode (HMDE) and PLA filament with carbon conductive admixture (PLA-C) for 3D printer. Contents of lead and cadmium in all analyzed bee venom samples were on the level of mg kg-1, of nickel and copper about ten times higher, and of zinc on the level of g kg-1. The results achieved using HMDE were recorded with average relative standard deviation (RSD) 5.4% (from 3.2% to 8.6%) and using PLA-C 11.8% (from 6.5% to 18.0%). The results achieved using both electrodes proved to be equivalent with statistical probability higher than 95%.