Molecular Identification of Enteroviruses from Cattle and Goat Feces and Environment in Thailand

Abstract

The identification and characterization of viruses of the genus Enterovirus in healthy and infected livestock, including cattle and goats, have been increasing. Enterovirus E (EV-E) and Enterovirus F (EV-F) are commonly found in cattle, whereas Enterovirus G (EV-G) is found in goats. In this study, molecular and phylogenetic analyses were performed to determine the prevalence of EVs in cattle and goat feces from Kanchanaburi Province, Thailand. The presence of EVs in water samples and the feces of other animals collected from the areas surrounding cattle and goat farms was also investigated. By use of 5=-untranslated region (5= UTR) real-time reverse transcription-PCR (RT-PCR), EVs were detected in 39.5% of cattle samples, 47% of goat samples, 35.3% of water samples, and one pool of chicken feces. Phylogenetic analysis revealed the presence of EV-E and EV-F in cattle, EV-E and EV-G in goats, and EV-F in water samples and chicken feces. Analysis of enteroviral VP1 sequences from cattle revealed that the EV-E genotypes circulating in the study region were EV-E1, with a possible new genotype that is closely related to EV-E2. Analysis of enteroviral VP1 sequences from goats suggested the circulation of EV-G5 and a possible new genotype that is closely related to EV-G20. Sequence analyses also suggested that although the VP1 sequences from goats were closely related to those of EV-G, which were considered porcine enterovirus sequences, their 5= UTRs form a separated cluster with sequences of sheep and goat origin, suggesting a new classification of the ovine/caprine-specific enterovirus group. IMPORTANCE Possible new EV-E and EV-G genotypes were identified for EVs detected in this study. The EV-E viruses were also successfully isolated from MDBK cells. The goat EV sequence analysis suggested the presence of an ovine/caprinespecific EV group that is different from EV-G of porcine origin. The significance of our research is that it identifies and characterizes possible novel EVs, thereby indicating that enteroviruses in animals are continually evolving. The facts that enteroviruses can persist in the environment, contaminate it for long periods, and be transmitted between animals raise serious concerns regarding this group of viruses as emerging livestock pathogens.

Evaluation of Cattle for Naturally Colonized Shiga Toxin-Producing Escherichia coli Requires Combinatorial Strategies

“…In contrast, no CPE was observed with fecal extracts from animal #5915, although the stx2 gene was amplified from the same sample ( Table 2 , Supplementary Figure 2 ), indicating a possibly nonfunctional gene. Additionally, non-Stx factors in the fecal extracts, such as viruses, may have caused CPE observed with 3/58 fecal extracts (animals #6044, #6069, and #6148) that were not neutralized with the anti-Stx antisera (Supplementary Figure 3 ) [ 77 , 78 ]. These variations indicate that Vero cell cytotoxicity assays require verification through neutralization steps and correlation with culture/PCR results.…”

Aichivirus C isolate is a diarrhoea‐causing pathogen in goats

“…Aichivirus C was detected by a RT‐PCR assay that targeted the 3D gene according to our previous report, with a limit at 2.87 × 10 3 copies per microliter (Abi et al., 2020). Peste des Petits Ruminants, rotavirus A, bovine coronavirus, bovine viral diarrhoea virus, caprine enterovirus, enterotoxigenic Escherichia coli K99 and Salmonella were detected by RT‐PCR or PCR assays, as described previously (Amine et al., 2020; Brichta‐Harhay et al., 2013; Gong et al., 2014; Income et al., 2019; Polci et al., 2015).…”

Molecular Analysis of Caprine Enterovirus Circulating in China during 2016–2021: Evolutionary Significance

“…Out of the 12 enterovirus species, EV-E and EV-F viruses infect cattle [ 2 , 3 , 4 , 5 , 6 , 7 ], while EV-G viruses generally infect pigs [ 8 , 9 , 10 , 11 ]. Recently, several enterovirus strains isolated from goats or sheep were also classified as EV-G [ 12 , 13 , 14 , 15 ]. Chang et al reported the caprine enterovirus strain SD-S67 isolated in cell culture from samples collected from goats as being EV-F, suggesting that goats are also natural hosts for EV-F [ 16 ].…”

“…Caprine/ovine enterovirus (CEV/OEV) infection has been increasingly reported in recent years [ 12 , 13 , 14 , 15 ]. However, the genetic variation and evolution of caprine enterovirus has scarcely been investigated.…”

“…Currently, the complete genome sequences of caprine/ovine enterovirus have been revealed only in very few strains [ 14 ]. The majority of the reported sequences of caprine/ovine enterovirus are either partial 5′ UTRs or a small portion of the coding sequence, such as VP1 genes [ 13 , 15 ]. Therefore, it is difficult to evaluate the genetic variation and evolution of caprine enterovirus, especially under the circumstance of the paucity of complete genome sequences.…”