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He G.-Z.,Guiyang College of Traditional Chinese Medicine | Deng S.-X.,Agricultural Office of Dalingshan Town
Journal of Animal and Veterinary Advances | Year: 2011

The objective of this study was to determine the replication kinetics of Salmonella enteritidis genome loads (S. enteritidis) in reproductive organs of laying duck after oral challenge. We conducted serovar-specific real-time PCR for S. enteritidis to detect the genomic DNA of S. enteritidis in laying duck at different time points. To validate these results, the Indirect Fluorescent Antibody (IFA) technique was employed too. The results showed that S. enteritidis was consistently detected in all the samples. However, the number of bacteria started decreasing by 3-4 days and by 6 days, the concentration of S. enteritidis DNA was below the detection limits of the PCR assay except the spleen and vagina. In conclusion, this study will help in understanding the pathogenesis of S. enteritidis infection in vivo. © Medwell Journals, 2011. Source


He G.-Z.,Guiyang College of Traditional Chinese Medicine | Deng S.-X.,Agricultural Office of Dalingshan Town
Journal of Poultry Science | Year: 2012

We used a real-time PCR for Salmonella Enteritidis to detect the genomic DNA of Salmonella Enteritidis live vaccine in the immune organs, including the bursa of Fabricius, thymus, spleen, and Harderian gland, from chicken after subcutaneously vaccinated at different time points. Significant numbers of Salmonella Enteritidis genomes in the immune organs were first detected at 12 hour (post-vaccination) p.v., and subsequently rose to peak levels during 48 h to 72 h p.v. The rapid early increase of vaccine levels in all samples examined followed by a steady decline from 84 h to 15 days p.v. The real-time PCR analysis of a variety of tissues is significant for further investigation of the mechanism of vaccinal protection, and the optimization of vaccination regimes. © 2012, Japan Poultry Science Association. Source


He G.-Z.,Guiyang College of Traditional Chinese Medicine | Feng Y.,Guiyang College of Traditional Chinese Medicine | Deng S.-X.,Agricultural Office of Dalingshan Town
Parasitology Research | Year: 2012

The objective of this study was to analyze the difference in intestinal microbial diversity between healthy and (Entamoeba histolytica) orally infected minipig. Enterobacterial repetitive intergenic consensus (ERIC)-PCR was applied to analyze this diversity and dynamic change, including the duodenum, jejunum, ileum, cecum, and rectum from healthy and orally infected minipig at different time points. The results showed that the intestinal microbial community of the control minipigs was stable and the ERIC-PCR band numbers of the control minipigs were the lowest in the rectum and highest in the cecum. ERIC-PCR bands of orally inoculated minipigs showed no obvious change until 24 h after postinoculation (hpi). The numbers of the ERIC-PCR bands gradually decreased from 24 to 72 hpi, then, with the development of disease, the band numbers gradually increased until 6 days postinoculation. The prominent bacteria had changed in the presence of E. histolytica infection and theDNAstar of staple of ERIC-PCR showed that obligate aerobes and facultative aerobes (Escherichia coli, Shigella, Salmonella) became the preponderant bacilli in the intestine of orally infected minipigs with E. histolytica. This study has provided significant data to clarify the intestinal microbial diversity and dynamic change of healthy and E. histolytica orally infected minipigs. © Springer-Verlag 2012. Source


He G.-Z.,Guiyang College of Traditional Chinese Medicine | Deng S.-X.,Agricultural Office of Dalingshan Town | Qian N.,Guiyang College of Traditional Chinese Medicine
Parasitology Research | Year: 2012

Enterobacterial repetitive intergenic consensus (ERIC)-PCR was applied to analyze the difference of intestinal microbial community diversity between healthy and orally infected rabbits with Entamoeba histolytica. The dynamic changes in different parts of digestive system including the duodenum, jejunum, ileum, caecum, and rectum in healthy and infected rabbits at different time points were also tested. The intestinal microbial community of the control healthy rabbits was steady, and the total number of ERIC-PCR bands in the control healthy rabbit was the least in the rectum and the most in the caecum. ERIC-PCR bands of orally inoculated rabbits did not obviously change until 24 h after postinoculation (p.i.). The numbers of the ERICPCR bands gradually decreased from 24 to 72 h p.i., and then, with the development of disease, the band numbers gradually increased until 6 days p.i. Sequence analysis showed that the nucleotide sequence homologies of the fragment about 1,200 bp of infected ileum sampled at 32 h p.i. were above 95 % with Sinorhizobium meliloti enterobacterial, Erwinia amylovora and Salmonella typhimurium, and the nucleotide sequence homologies of the fragment about 300 bp of infected ileum sampled at 48 h p.i. were more than 90%with Xanthomonas campestris enterobacterial, Yersinia enterocolitica subsp., Shigella flexneri, S. meliloti enterobacterial, Yersinia pestis, Klebsiella pneumoniae subsp., and Escherichia coli. The prominent bacteria had changed after E. histolytica infection. The DNAstar of staple of ERICPCR showed that aerobe and facultative aerobe (E. coli, Shigella, and Salmonella) became preponderant bacilli in the intestine of orally infected rabbits with E. histolytica. ©Springer-Verlag 2012. Source


He G.Z.,Guiyang College of Traditional Chinese Medicine | Deng S.X.,Agricultural Office of Dalingshan Town | Tian W.Y.,Guiyang College of Traditional Chinese Medicine
Iranian Journal of Veterinary Research | Year: 2013

Serovar-specific real-time PCR for Salmonella enterica serovar Enteritidis (S. Enteritidis) was conducted to detect the genomic DNA of S. Enteritidis from laying goose after subcutaneous vaccination at different time points. Indirect fluorescent antibody (IFA) technique and immunohistochemical localization were employed to validate the results. The results showed that S. Enteritidis was consistently detected in all the samples. Vagina and uterus were positive at 20 h PI, and the last organ to show a positive result was the largest and third largest preovulatory follicle, at 32 h PI. The copy numbers of S. Enteritidis DNA in each tissue reached a peak at 36-60 h PI, with the vagina and uterus containing higher concentrations than other tissues. However, the number of bacteria started decreasing by 3-4 d, and by 6 d, the concentration of S. Enteritidis DNA was below the detection limits of the PCR assay, except the vagina. The real-time PCR analysis of a variety of tissues is significant for further investigation of the mechanism of vaccine protection and the optimization of vaccination regimes. Source

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