Background:Diarrhea is a major cause of morbidity and mortality in young calves,resulting in considerable economic loss for dairy farms.To determine if some gut microbes might have resistance to dysbiotic process with...Background:Diarrhea is a major cause of morbidity and mortality in young calves,resulting in considerable economic loss for dairy farms.To determine if some gut microbes might have resistance to dysbiotic process with calf diarrhea by dictating the microbial co-occurrence patterns from birth to post-weaning,we examined the dynamic development of the gut microbiota and diarrhea status using two animal trials,with the first trial having 14 Holstein dairy calves whose fecal samples were collected 18 times over 78 d from birth to 15 d post-weaning and the second trial having 43 Holstein dairy calves whose fecal samples were collected daily from 8 to 18 days of age corresponding to the first diarrhea peak of trial 1.Results:Metataxonomic analysis of the fecal microbiota showed that the development of gut microbiota had three age periods with birth and weaning as the separatrices.Two diarrhea peaks were observed during the transition of the three age periods.Fusobacteriaceae was identified as a diarrhea-associated taxon both in the early stage and during weaning,and Clostridium_sensu_stricto_1 was another increased genus among diarrheic calves in the early stage.In the neonatal calves,Prevotella_2(ASV4 and ASV26),Prevotella_9(ASV43),and Alloprevotella(ASV14)were negatively associated with Clostridium_sensu_stricto_1(ASV48),the keystone taxa of the diarrhea-phase module.During weaning,unclassified Muribaculaceae(ASV28 and ASV44),UBA1819(ASV151),Barnesiella(ASV497),and Ruminococcaceae_UCG-005(ASV254)were identified being associated with non-diarrheic status,and they aggregated in the non-diarrhea module of co-occurrence patterns wherein unclassified Muribaculaceae(ASV28)and Barnesiella(ASV497)had a direct negative relationship with the members of the diarrhea module.Conclusions:Taken together,our results suggest that the dynamic successions of calf gut microbiota and the interactions among some bacteria could influence calf diarrhea,and some species of Prevotella might be the core microbiota in both neonatal and weaning 展开更多
A real-time RT-PCR assay using Taq Man-MGB probes was developed to detect and type the bovine viral diarrhea virus（BVDV） in cattle.Universal primers and Taq Man-MGB probes were designed from the 5′-untranslated reg...A real-time RT-PCR assay using Taq Man-MGB probes was developed to detect and type the bovine viral diarrhea virus（BVDV） in cattle.Universal primers and Taq Man-MGB probes were designed from the 5′-untranslated region of known pestiviral sequences.Prior to optimizing the assay, c RNAs were transcribed in vitro from the BVDV 1 and BVDV 2 RTPCR products to make standard curves.The detection limit of the assay was 1.72×102 copies for BVDV 1 and 2.14×102copies for BVDV 2.The specificity of the assay evaluated on several BVDV strains including bovine herpesvirus 1（BHV 1）, foot and mouth disease virus（FMDV） and several classical swine fever virus（CSFV） strains showed specific detection of the positive virus over 40 cycles.The assay was highly reproducible with the coefficient of variance ranging from 1.04 to 1.33% for BVDV 1 and from 0.83 to 1.48% for BVDV 2, respectively.Using this method, we tested a total of 2 327 cattle from three dairy farms for the presence of BVDV persistently infected（PI） animals.In this assay, each RT-PCR template contained a mixture of ten samples from different animals.The occurrence rate of PI cattle in three farms ranging from 0.9 to 2.54% could represent partly the PI rates in cattle farm in China.In conclusion, using our real-time PCR assay, we could effectively detect and type BVDV and identify PI cattle in a rapid and cost-effective manner.展开更多
基金the National Key Research and Development Program of China(2017YFD0500502)。
文摘Background:Diarrhea is a major cause of morbidity and mortality in young calves,resulting in considerable economic loss for dairy farms.To determine if some gut microbes might have resistance to dysbiotic process with calf diarrhea by dictating the microbial co-occurrence patterns from birth to post-weaning,we examined the dynamic development of the gut microbiota and diarrhea status using two animal trials,with the first trial having 14 Holstein dairy calves whose fecal samples were collected 18 times over 78 d from birth to 15 d post-weaning and the second trial having 43 Holstein dairy calves whose fecal samples were collected daily from 8 to 18 days of age corresponding to the first diarrhea peak of trial 1.Results:Metataxonomic analysis of the fecal microbiota showed that the development of gut microbiota had three age periods with birth and weaning as the separatrices.Two diarrhea peaks were observed during the transition of the three age periods.Fusobacteriaceae was identified as a diarrhea-associated taxon both in the early stage and during weaning,and Clostridium_sensu_stricto_1 was another increased genus among diarrheic calves in the early stage.In the neonatal calves,Prevotella_2(ASV4 and ASV26),Prevotella_9(ASV43),and Alloprevotella(ASV14)were negatively associated with Clostridium_sensu_stricto_1(ASV48),the keystone taxa of the diarrhea-phase module.During weaning,unclassified Muribaculaceae(ASV28 and ASV44),UBA1819(ASV151),Barnesiella(ASV497),and Ruminococcaceae_UCG-005(ASV254)were identified being associated with non-diarrheic status,and they aggregated in the non-diarrhea module of co-occurrence patterns wherein unclassified Muribaculaceae(ASV28)and Barnesiella(ASV497)had a direct negative relationship with the members of the diarrhea module.Conclusions:Taken together,our results suggest that the dynamic successions of calf gut microbiota and the interactions among some bacteria could influence calf diarrhea,and some species of Prevotella might be the core microbiota in both neonatal and weaning
基金supported by the China Agriculture Research System(CARS-37)
文摘A real-time RT-PCR assay using Taq Man-MGB probes was developed to detect and type the bovine viral diarrhea virus（BVDV） in cattle.Universal primers and Taq Man-MGB probes were designed from the 5′-untranslated region of known pestiviral sequences.Prior to optimizing the assay, c RNAs were transcribed in vitro from the BVDV 1 and BVDV 2 RTPCR products to make standard curves.The detection limit of the assay was 1.72×102 copies for BVDV 1 and 2.14×102copies for BVDV 2.The specificity of the assay evaluated on several BVDV strains including bovine herpesvirus 1（BHV 1）, foot and mouth disease virus（FMDV） and several classical swine fever virus（CSFV） strains showed specific detection of the positive virus over 40 cycles.The assay was highly reproducible with the coefficient of variance ranging from 1.04 to 1.33% for BVDV 1 and from 0.83 to 1.48% for BVDV 2, respectively.Using this method, we tested a total of 2 327 cattle from three dairy farms for the presence of BVDV persistently infected（PI） animals.In this assay, each RT-PCR template contained a mixture of ten samples from different animals.The occurrence rate of PI cattle in three farms ranging from 0.9 to 2.54% could represent partly the PI rates in cattle farm in China.In conclusion, using our real-time PCR assay, we could effectively detect and type BVDV and identify PI cattle in a rapid and cost-effective manner.
