Interspecies transmission of SARS CoV-2 with special emphasis on viral mutations and ACE-2 receptor homology roles

ABSTRACTCOVID-19 outbreak was first reported in 2019, Wuhan, China. The spillover of the disease caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), to a wide range of pet, zoo, wild, and farm animals has emphasized potential zoonotic and reverse zoonotic viral transmission. Furthermore, it has evoked inquiries about susceptibility of different animal species to SARS-CoV-2 infection and role of these animals as viral reservoirs. Therefore, studying susceptible and non-susceptible hosts for SARS-CoV-2 infection could give a better understanding for the virus and will help in preventing further outbreaks. Here, we review structural aspects of SARS-CoV-2 spike protein, the effect of the different mutations observed in the spike protein, and the impact of ACE2 receptor variations in different animal hosts on inter-species transmission. Moreover, the SARS-CoV-2 spillover chain was reviewed. Combination of SARS-CoV-2 high mutation rate and homology of cellular ACE2 receptors enable the virus to transcend species barriers and facilitate its transmission between humans and animals

Major Gram-negative bacterial causes isolated from apparent Healthy and diarrheic foals in Egypt, prevalence, identification and antibiotic susceptibility profiles

Diarrhea is one of the most significant illnesses affecting young foals and may be manifested in more than half of foals up to 3 months of age. Numerous infectious causes have been involved but bacterial pathogens are concerned. This study aimed to assess and identify the prime Gram-negative bacteriological etiology of Arabian horse foals’ diarrhea in Egypt and to designate the antibiotic susceptibility profiles of the isolated microorganisms.  Rectal swabs and internal organs were obtained from 216 foals (89 diarrheic and 127 apparently healthy), aged 1 week to 1 year old, reared in Arabian horse farms in Great Cairo, Egypt during a period extended from March 2022 to December 2022. Conventional bacteriological examination was performed using selective media persuaded by routine and advanced biochemical tests. The isolation displayed 648 bacterial isolates; 452 (68.6%) were Gram-negative with the mixed isolation representing about 28%. Escherichia coli constituted the most prevalent; 110 / 452 (24.34%); 65 in apparently healthy foals and 45 among diarrheic ones. The second was Klebsiella pneumoniae (72, 15.92%) at which 50, (15.67%) were found to expose the highest causative agent recovered from diarrheic foals. Imipenem, quinolones, and trimethoprim/sulfamethoxazole were the most effective versus Gram-negative species isolated from diarrheic foals while ampicillin and tetracycline were of no effect. The phenotypic antibiotic susceptibility test revealed a picture of multidrug resistance (MDR) as 111 isolates (28.71%) showed resistance to three or more antibiotics belonging to different groups. The results demonstrated that Klebsiella pneumonia was the highest MDR species (54.16%) followed by Pseudomonas aeruginosa, Salmonella enterica, and Escherichia coli isolates by 43.9, 32.1 and 30.9 % respectively. In conclusion, Gram-negative bacteria constituted the major causative agents of diarrhea in the Arabian foals. The most effective antibacterial drugs were imipenem, quinolones, and trimethoprim/ sulfamethoxazole. The issue of the existence of multidrug resistance isolates should be considered for proper therapy of foal diarrhea

Advanced molecular characterization of enteropathogenic Escherichia coli isolated from diarrheic camel neonates in Egypt

Background and Aim: Camels are important livestock in Egypt on cultural and economic bases, but studies of etiological agents of camelid diseases are limited. The enteropathogen Escherichia coli is a cause of broad spectrum gastrointestinal infections among humans and animals, especially in developing countries. Severe infections can lead to death. The current study aimed to identify pathogenic E. coli strains that cause diarrhea in camel calves and characterize their virulence and drug resistance at a molecular level. Materials and Methods: Seventy fecal samples were collected from diarrheic neonatal camel calves in Giza Governorate during 2018-2019. Samples were cultured on a selective medium for E. coli, and positive colonies were confirmed biochemically, serotyped, and tested for antibiotic susceptibility. E. coli isolates were further confirmed through detection of the housekeeping gene, yaiO, and examined for the presence of virulence genes; traT and fimH and for genes responsible for antibiotic resistance, ampC, aadB, and mphA. The isolates in the important isolated serotype, E. coli O26, were examined for toxigenic genes and sequenced. Results: The bacteriological and biochemical examination identified 12 E. coli isolates from 70 fecal samples (17.1%). Serotyping of these isolates showed four types: O26, four isolates, 33.3%; O103, O111, three isolates each, 25%; and O45, two isolates, 16.7%. The isolates showed resistance to vancomycin (75%) and ampicillin (66.6%), but were highly susceptible to ciprofloxacin, norfloxacin, and tetracycline (100%). The structural gene, yaiO (115 bp), was amplified from all 12 E. coli isolates and traT and fimH genes were amplified from 10 and 8 isolates, respectively. Antibiotic resistance genes, ampC, mphA, and aadB, were harbored in 9 (75%), 8 (66.6%), and 5 (41.7%), respectively. Seven isolates (58.3%) were MDR. Real-time-polymerase chain reaction of the O26 isolates identified one isolate harboring vt1, two with vt2, and one isolate with neither gene. Sequencing of the isolates revealed similarities to E. coli O157 strains. Conclusion: Camels and other livestock suffer various diseases, including diarrhea often caused by microbial pathogens. Enteropathogenic E. coli serotypes were isolated from diarrheic neonatal camel calves. These isolates exhibited virulence and multiple drug resistance genes

Major Gram-negative bacterial causes isolated from apparent Healthy and diarrheic foals in Egypt, prevalence, identification and antibiotic susceptibility profiles

Diarrhea is one of the most significant illnesses affecting young foals and may be manifested in more than half of foals up to 3 months of age. Numerous infectious causes have been involved but bacterial pathogens are concerned. This study aimed to assess and identify the prime Gram-negative bacteriological etiology of Arabian horse foals’ diarrhea in Egypt and to designate the antibiotic susceptibility profiles of the isolated microorganisms.  Rectal swabs and internal organs were obtained from 216 foals (89 diarrheic and 127 apparently healthy), aged 1 week to 1 year old, reared in Arabian horse farms in Great Cairo, Egypt during a period extended from March 2022 to December 2022. Conventional bacteriological examination was performed using selective media persuaded by routine and advanced biochemical tests. The isolation displayed 648 bacterial isolates; 452 (68.6%) were Gram-negative with the mixed isolation representing about 28%. Escherichia coli constituted the most prevalent; 110 / 452 (24.34%); 65 in apparently healthy foals and 45 among diarrheic ones. The second was Klebsiella pneumoniae (72, 15.92%) at which 50, (15.67%) were found to expose the highest causative agent recovered from diarrheic foals. Imipenem, quinolones, and trimethoprim/sulfamethoxazole were the most effective versus Gram-negative species isolated from diarrheic foals while ampicillin and tetracycline were of no effect. The phenotypic antibiotic susceptibility test revealed a picture of multidrug resistance (MDR) as 111 isolates (28.71%) showed resistance to three or more antibiotics belonging to different groups. The results demonstrated that Klebsiella pneumonia was the highest MDR species (54.16%) followed by Pseudomonas aeruginosa, Salmonella enterica, and Escherichia coli isolates by 43.9, 32.1 and 30.9 % respectively. In conclusion, Gram-negative bacteria constituted the major causative agents of diarrhea in the Arabian foals. The most effective antibacterial drugs were imipenem, quinolones, and trimethoprim/ sulfamethoxazole. The issue of the existence of multidrug resistance isolates should be considered for proper therapy of foal diarrhea

Development of multiplex gold nanoparticles biosensors for ultrasensitive detection and genotyping of equine herpes viruses

Abstract Gold nanoparticles (GNPs) biosensors can detect low viral loads and differentiate between viruses types, enabling early diagnosis and effective disease management. In the present study, we developed GNPs biosensors with two different capping agent, citrate-GNPs biosensors and polyvinylpyrrolidone (PVP)-GNPs biosensors for detection of EHV-1 and EHV-4 in multiplex real time PCR (rPCR). Citrate-GNPs and PVP-GNPs biosensors can detect dilution 1010 of EHV-1 with mean Cycle threshold (Ct) 11.7 and 9.6, respectively and one copy as limit of detection, while citrate-GNPs and PVP-GNPs biosensors can detect dilution 1010 of EHV-4 with mean Ct 10.5 and 9.2, respectively and one copy as limit of detection. These findings were confirmed by testing 87 different clinical samples, 4 more samples were positive with multiplex GNPs biosensors rPCR than multiplex rPCR. Multiplex citrate-GNPs and PVP-GNPs biosensors for EHV-1 and EHV-4 are a significant breakthrough in the diagnosis of these virus types. These biosensors offer high sensitivity and specificity, allowing for the accurate detection of the target viruses at very low concentrations and improve the early detection of EHV-1 and EHV-4, leading to faster control of infected animals to prevent the spread of these viruses

Isolation and Genotypic Characterization of New Emerging Avian Reovirus Genetic Variants in Egypt

Avian reovirus (ARV) strains cause a variety of symptoms in chickens, including viral arthritis/tenosynovitis, a disease that has emerged as a significant cause of economic losses in commercial chicken flocks in recent years in various countries, including Egypt. Furthermore, ARV strains are frequently isolated from birds suffering from malabsorption. In the actual study, seventy-five samples were collected in 2021 and 2022 from broiler and vaccinated broiler breeder flocks at different farms in Giza Province, Egypt, with reovirus-like symptoms such as significant weight fluctuation and arthritis/malabsorption. ARV was screened using real-time PCR, and fifteen positive samples were detected (20%), which were then subjected to embryonated chicken egg (ECE) isolation and molecular characterization (11/15 sample) of a partial segment of the sigma (σ)C gene (S1-gene). Phylogenetically, nine strains were found to belong to genotypic cluster IV, with 82–89% identity with Israeli ARV 2018, and two strains belong to genotypic cluster V with a 78% nucleotide identity with Japan ARV 2021. No correlation between lesions and genotype was found. The strains under study had a low sequence identity (43–55%) when compared with various commercial vaccines belonging to genotypic cluster I (e.g., strain S1133). These findings imply that novel ARV genotypes representing clusters IV and V have recently been introduced to Egyptian poultry farms. A homologous vaccine is suggested; because this variation raises the possibility that commercial vaccines may not offer protection against circulating ARVs

SARS‐CoV‐2 infection of companion animals in Egypt and its risk of spillover

Abstract Background Reverse zoonoses occur because of interactions between humans and animals. Homology of ACE‐2 cell receptors in different hosts and high mutation rate of SARS‐CoV‐2 enhance viral transmission among species. Objectives This study aimed to investigate spillover of SARS‐CoV‐2 between humans and companion animals. Methods A cross‐sectional study was constructed using nasopharyngeal/oropharyngeal swabs, serum and blood samples collected from 66 companion animals (33 cats and 33 dogs) that were in contact with SARS‐CoV‐2‐positive owners from December 2020 to March 2021. Swabs were screened by rRT‐PCR and some positive cases were confirmed by partial spike gene sequencing. Clinical pathology and pathological studies were also performed. Results Our findings revealed that 30% of cats (10/33) and 24% of dogs (8/33) were SARS‐CoV‐2 positive. While 33% of these animals were asymptomatic (6/18), 28% showed mild respiratory signs (5/18) and 39% displayed severe respiratory signs (7/18) including 4 dead cats 40% (4/10). Partial spike gene sequencing of 6 positive samples collected in December 2020 were identical to SARS‐CoV‐2 that was detected in humans in Egypt in that time frame. Clinical pathology findings revealed thrombocytopenia, lymphocytopenia, as well as elevated levels of D‐dimer, LDH, CRP, and ferritin. Post‐mortem and histopathological examinations illustrated multisystemic effects. Conclusions There is a potential occurrence of SARS‐CoV‐2 spillover between humans and pet animals. Impacts The present study highlighted the potential occurrence of SARS‐CoV‐2 spillover between humans and their companion animals. Biosecurity measures should be applied to decrease spread of SARS‐CoV‐2 among humans and pet animals

Cross Sectional, Seroprevalence Study of Peste des Petits Ruminants and the Related Risk Factors During Outbreak in Goats’ Farm in Egypt

Peste des Petits ruminants' virus (PPRV) is a notifiable transboundary and economically significant viral disease that affects goats and sheep. The current study was conducted to identify the seroprevalences of PPRV in goat farm in Marsa-Matroh province, Egypt during an outbreak in 2022. Moreover, this work aims to study the relevant risk factors directly related to the virus infection and attempt molecular characterization and phylogenetic analysis of the circulating strain. The samples included 356 sera, 154 nasal swabs, and 10 tissue samples were collected for PPRV screening and molecular characterization. The seroprevalence percent was found of 42.69%.  It was observed that the prevalence rate, and case-fatality rate were higher under 6 months of age than in adults, and referring to sex, females had a more significant disease incidence than males. Based on phylogenetic analysis; the strains of the current study: PPR/AHRI-Matrouh1/Egy/2022 (accession number: OP881991) and PPR/AHRI-Matrouh2/Egy/2022 (accession number: OP881992) were identified as PPRV lineage IV, with 99.2% and 98.8% identity to the Ethiopian strain (Accession number MK571524) and Sudanese strain (Accession no HQ131931) respectively. An efficient PPR vaccination program with rigorous quarantine measures at the borders is advised to be implemented in the country to control the spread of the disease and avoid the entry of novel strains into the Egyptian governorates

Antigenic evaluation of extracted fimbrial protein obtained from pathogenic Escherichia coli isolated from diarrheic camel neonates

Enteropathogenic Escherichia coli (EPEC) were considered prime reason of diarrhea among neonatal livestock animals in developing countries and could be of public health importance via contaminated milk and meat.  Continual attempts were conducted to combat this illness using various antigenic determinants. This study was performed on four E. coli serotypes O26, O45, O103 and O111, which were previously recovered from diarrheic camel calves in Giza, Egypt. Extraction of fimbrial proteins was carried out through dialysis then evaluation of their immunogenic activity was preceded. SDS-PAGE electrophoresis was performed on crude extracted fimbrial proteins and revealed single band for each isolate ranged from 22 to 33kDa. Immunobloting was implemented on the extracted crude fimbrial proteins against E.coli O26 antisera formerly prepared in rabbit.These findings suggested that the fimbrial proteins are of immunogenicity importance and can serve as a protective passive vaccine antigen in prevention of diarrhea caused by EPEC and ETEC infection in camel calves

Molecular Characterization of Newly Emerging Foot-and-Mouth Disease Virus Serotype SAT 2 of Lib-12 Lineage Isolated from Egypt

An outbreak of foot-and-mouth disease virus (FMDV) serotype SAT 2 occurred in Egypt in 2018, which affected cattle and water buffalo. Previous phylogenetic studies on FMDV circulating in Egypt have mainly focused on genomic regions encoding structural proteins which determine FMDV serotype. So far, none of these studies have analyzed the open reading frame (ORF) sequence of Egyptian SAT 2/Lib-12 lineage. The present study aimed to analyze and identify the ORF genome sequence of Lib-12 lineage which belongs to FMDV serotype SAT 2 topotype VII in Egypt. The protocol workflow was optimized and tested using a representative field isolate of FMDV/ SAT 2/Lib-12 from a bovine tongue sample collected in 2018 from Ismailia governorate (SAT2/EGY/Ismailia/ 2018). The protocol was based on reverse transcription polymerase chain reaction with multiple overlapping primers, amplicons sequencing, and assembly to complete the ORF consensus sequence. Alignments of the sequence fragments formed consensus genome sequence of 7219 nucleotides in length. The complete nucleotide sequence of the Egyptian isolate was related to Ethiopian, Nigerian, and Ghanaian strains, with identity not exceeding 95%. The divergence in the genetic identity of the Egyptian SAT 2/Lib-12 lineage from other Egyptian strains and Libyan isolates was 7%, and this may be attributed to the absence of the Lib-12 lineage ORF sequence from Egypt and Libya in the database. The present study significantly advances knowledge of the molecular analysis of FMDV SAT 2 and the design of vaccine selection for FMDV SAT 2 in Egypt. The study protocol could be applied to other FMDV serotypes