Diarrhoeic Kids Research Articles

Genetic and phylogenetic relationship of an American caprine rotavirus B strain with equine rotavirus B

Rotaviruses (RVs) are significant enteric pathogens of humans and animals. In March 2021, the University of Kentucky Veterinary Diagnostic Laboratory (UKVDL) received a fecal sample from a 1-week-old goat kid with diarrhea from a farm with 5 additional diarrheic kids. The fecal sample was tested negative for Bovine coronavirus, Bovine rotavirus Group A, E. coli K99+, Cryptosporidium parvum and Salmonella spp by multiplex real-time PCR assays. Interestingly, a novel Equine Rotavirus B (ERVB) in Kentucky was identified from neonatal foals also with watery diarrhea in the Spring of 2021. Once the ERVB-specific real-time PCR assay became available, the fecal sample from the goat kid was tested and found positive for RVB. Genome sequence of the caprine RVB from fecal sample was obtained using shotgun metagenomic sequencing by Illumina MiSeq. All of the eleven viral segments of caprine RVB were sequenced either completely or partially. Genetic and phylogenetic analysis of VP7, VP4, VP6, VP1–VP3, and NSP1- NSP5 genes indicated the caprine RVB strain had the genotype constellation of G3-P[3]-I3- R3-C3-M3-A4-N3-T3-E3-H3. In addition, the caprine RVB sequence showed the highest nucleotide identity and evolutionary relationship to the ERVB and previously sequenced caprine RVB strains. Given the similar geographical location of the equine and caprine strains from our study, these findings suggest a possible common source of infection.

New Insights on Coding Mutations and mRNA Levels of Candidate Genes Associated with Diarrhea Susceptibility in Baladi Goat

The purpose of this investigation was to examine mutations and mRNA levels of potential genes linked to diarrhea susceptibility in order to assess the health status of diarrheic kids of Baladi goats. One hundred female Baladi kids (35 diarrheic and 65 apparently healthy) were used. PCR-DNA sequencing was conducted for TMED1, CALR, FBXW9, HS6ST3, SMURF1, KPNA7, FBXL2, PIN1, S1PR5, ICAM1, EDN1, MAPK11, CSF1R, LRRK1, and CFH markers revealed nucleotide sequence variants in the frequency of distribution of all detected SNPs (p ˂ 0.05) between healthy and affected kids. Chi-square analysis showed a significant difference between resistant and affected animals. Gene expression profile revealed that TMED1, CALR, FBXW9, HS6ST3, SMURF1, KPNA7, FBXL2, PIN1, S1PR5, ICAM1, EDN1, MAPK11, CSF1R and LRRK1 were significantly up-regulated in diarrheic kids than resistant ones. Meanwhile, CFH gene elicited an opposite trend. On the mRNA levels of the examined indicators, there was a substantial interaction between the type of gene and diarrhea resistance/susceptibility. The findings could support the importance of nucleotide variations and the expression pattern of the examined genes as biomarkers for diarrhea resistance/susceptibility and offer a useful management strategy for Baladi goats.

Detection and molecular characterization of kobuvirus from diarrheic goats in Minnesota.

Kobuvirus infections are common among humans, rodents, carnivores, pigs, and ruminants. We report herein the complete genome sequence of a novel caprine kobuvirus (MN604700) from diarrheic kids in Minnesota. Whole-genome sequencing revealed a kobuvirus genome of 8,139 nt with a single ORF region encoding a polyprotein of 2,480 amino acids. Further analysis revealed nt substitutions along the genome compared with that of the caprine kobuvirus reference strain, with 93% identity. Phylogenetic analysis indicated that the clade of the caprine kobuvirus was most closely related to porcine kobuviruses rather than bovine or ovine kobuviruses. Using primers designed from this genome, caprine kobuvirus was identified in the stools of other goats. Sanger sequencing of PCR products indicated 3D and VP1 gene nucleotides of this latter strain were 95% and 91% identical with those of MN604700, respectively. There were 35 and 101 nt substitutions in 3D and VP1 genes, respectively. Findings of kobuvirus over a 2-y period may indicate an endemic state, which needs further research. In addition, screening for kobuviruses over large geographic areas is needed to identify the evolutionary connections among different strains.

Dynamic Distribution of Gut Microbiota in Goats at Different Ages and Health States.

The importance of the gut microbiota (GM) of animals is widely acknowledged because of its pivotal roles in metabolism, immunity, and health maintenance. The level of health can be reflected by the dynamic distribution of GM. In this study, high-throughput sequencing of the bacterial 16S rRNA gene was used to compare the microbial populations from feces in healthy and diarrheic kids, which reflected the dynamic shift of microbiota in kids and investigated differences from adult healthy goats. Healthy kids and goats not only displayed higher species richness but also exhibited higher bacterial diversity than diarrheic kids based on the results of the operational taxonomic unit analysis, alpha diversity, and beta diversity. Firmicutes and Bacteroidetes were the most dominant phyla in all samples. At the genus level, the differences in diversity and abundance between diarrheic kids and the other two groups were gradually observed. In the diarrheic kid intestine, Bacteroides remained the dominant species, and the proportion of Clostridium_sensu_stricto_1 and Paeniclostridium increased, whereas Rikenellaceae_RC9_gut_group, Ruminococcaceae_UCG-005, and Christensenellaceae_R-7_group were significantly reduced. The results showed the differences of GM in diarrheic kids and healthy kids were significant while in kids and goats were not obvious. Differences in the composition of intestinal microbiota may not be the cause of diarrhea, and some changes of bacterial richness may guide our interpretation of diarrhea. This study is the first to investigate the distribution of GM in Boer goats with different ages and health states. Furthermore, this study will provide a theoretical basis for the establishment of a prevention and treatment system for goat diarrhea.

Prevalence and molecular characterisation of Cryptosporidium spp. in goat kids

Prevalence and molecular characterisation of Cryptosporidium species was done in kids belonging to organised and non-organised goat farms at Jabalpur. The overall prevalence of Cryptosporidium was 14.63%. The prevalence was non-significantly higher in male kids (16.16%) as compared to that of female kids (13.21%). Age wise prevalence was higher in kids up to one month age (16.13%) than that of kids upto 3 months age (13.99%). No significant difference was found in prevalence among different breeds and in kids kept in farm or field conditions. The prevalence was non-significantly higher in non-diarrhoeic kids than diarrhoeic kids. Most of the infections were of one score (76.6%). Molecular characterisation by PCR-RFLP of 18S SSU rRNA gene revealed presence of Cryptosporidium parvum species in positive faecal samples.

Detection of Clostridium perfringens toxinotypes, enteropathogenic E. coli, rota and corona viruses in the intestine of neonatal goat kids by molecular techniques

The role of various pathogenic agents affecting gastro-intestinal tract of neonatal goat kids was studied. Intestinal tissue samples of 142 necropsied kids died due to enteritis, pneumo-enteritis, tapeworm’s infestations and septicemia were collected for detection of toxinotypes of Clostridium perfringens, enteropathogenic E. coli (EPEC), Group-A rotavirus (GARV) and bovine coronavirus (BCV). C. perfringens toxinotyping was done by multiplex PCR using primers for cpa, cpb, cpb2, etx and iap gene. Further, identification of EPEC was done using SYBR green based real-time PCR assay targeting bfpA gene. Detection of GARV and BCV was done by one-step RT-PCR (osRTPCR). The incidence of C. perfringens was 16.20% (47.83% toxinotype A and 52.17% toxinotype D), with b2- toxin present in 30.43% of the samples. Incidence of EPEC in 0-1 and 1–3 month diarrhoeic kids was 36.62% and 25.35%, respectively. For viral aetiology, 11.97% were positive for GARV and 9.86% positive for BCV. Mixed infection of C. perfringens and EPEC was seen in 12.68% cases, while it was 2.11% for C. perfringens + GARV and 1.40% for C. perfringens + BCV. Similarly, the mixed occurrence of EPEC+GARV was 10.56% and that of EPEC+BCV was 7.04%. The combined infection of EPEC+GARV+BCV was 0.70%. The results suggested that C. perfringens type A and type D are the common toxinotypes affecting neonatal goat kids, with β2 toxin being an additional potential virulence factor. SYBR green based real-time PCR assay can be used as quick lab-based technique for efficient screening of EPEC from enteritis affected kids.Group-A rotavirus and bovine coronavirus appeared to be instrumental in causing mixed infection enteritis in kids.