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Lymphocyte Apoptosis in Vivo induced by Duck Swollenhead Hemorrhagic disease Virus

Lymphocyte Apoptosis in Vivo induced by Duck Swollenhead Hem...

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第四届第九次全国学术研讨会暨饲料和动物源食品安全战略论坛

作者： Li Chuanfeng Cheng Anchun Wang Mingshu Shen Chanjuan Zhang Na A Zhou Yi Chen Xiaoyue (Avian disease Research Center,College of Veterinary Medicine,sichuan Agricultural University,yaan,sichuan 625014 China key laboratory of animal disease and human health of sichuan province,yaan,sichuan 625014,China)

We studied apoptosis induced by duck swollenhead hemorrhagic disease virus (DSHDV) in vivo,focusing on the lymphoid organs that constitute the main targets for infection:bursa of Fabricius (BF), thymus,and spleen.72 ducks were inoculated intramuscularly with a virulent strain of *** morphology of lymphoid organs of these infected ducks was observed by light microscopy and transmission electron *** apoptosis was observed in DSHDV-infected BF,thymus, and *** apoptotic peak appeared at 72h *** death by classic necrosis was also observed in dead ducks after 72h postinoculation.,and it was further confirmed by in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL).We conclude in DSHDV-infected ducks that apoptosis of lymphocytes resulted in the depletion of iymphocytes,may play an important role in the pathogenesis of duck viral swollenhead hemorrhagic disease.

关键词： duck swollenhead hemorrhagic disease virus apoptosis necrosis lymphocyte

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The quantity influence of Bifidobacterium, Bacillus,and enterococcus in digestive tract of Duck Immunized with Low Duck Plague Virus by Oral Innoculation

The quantity influence of Bifidobacterium, Bacillus,and ente...

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第三届第八次全国学术研讨会暨动物微生态企业发展战略论坛

作者： HUANG Yongcheng,CHENG Anchun,WANG Mingshu,YANG Xiaoyan, QI Xuefeng, GE Zhongyuan,GUO Yufei,ZHANG Suhui,HUji,CHEN Xiaoyue (Avian disease Research Center, College of animal Science and Veterinary Medicine of sichuan Agricultural University key laboratory of animal disease and human health of sichuan province, yaan, sichuan, China,625014)

The article uses a pair of Bifidobacterium, the bacillus and the enterococcus FQ-PCR examination methods which have established, to detect a DPV weak poisonous vaccine oral administration immunity poult duck’s digestive tract pair of Bifidobacterium bifidus, the bacillus and the enterococcus carries’ change of quantity. The bacillus bifidus’ quantity change is:The content of Esophagus and rectum’s each time section separately close with control group’s, after immunity vaccination the duodenum, the jejunum, the ileum, the caecum’ contents increase 10 times compared with the control group some time section. The bacillus’ quantity change is: the content of Duodenum and rectum close with control group’s;The change of the esophagus and the jejunum’s content compared with the control group assume certain undulatory character, but have not obviously difference;Ileum and caecum’s content compared to control group’s many 10 times, these time section mainly distributes separately in 48h, 72h, 6d, 12d, 21d. The enterococcus quantity change is: The duodenum, the ileum and the rectum’s content change compared to the control group compare assume certain undulatory character, but have not obvious difference;Jejunum’s content in some time section compared to control group’s many 10 times;Caecum’s content in some time section compared to control group’s few 10 times. The purpose of this research is expounded the relation between the DPV weak poison vaccine and the digestive tract micro ecological association as well as to provided the foundation data for the immunity mechanism of the DPV weak poisonous vaccine.

关键词： Low Duck Plague Virus Oral administration Duck Bifldobacterium Bacillus Enterococcus

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Application of 16S rDNA Real-time Fluorescent Quantitate PCR for Detecting Gamma Germ

Application of 16S rDNA Real-time Fluorescent Quantitate PCR...

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第三届第八次全国学术研讨会暨动物微生态企业发展战略论坛

作者： HUANG Yongcheng,CHENG Anchun,WANG Mingshu,YANG Xiaoyan,QI Xuefeng, GE Zhongyuan,GUO Yufei,ZHANG Suhui,HUji,CHEN Xiaoyue (Avian disease Research Center, College of animal Science and Veterinary Medicine of sichuan Agricultural University key laboratory of animal disease and human health of sichuan province, yaan, sichuan, China,625014)

Group-specificity quantitate PCR primers and probe were designed and synthesized ,and method of detecting Bacillus was *** optimal Mg2+ concentration of the real-time fluorescent quantitate PCR was 7.5 mmol/L, and the optimal probe concertration was 0.1 μmol/L;The detecting of Bacillus pumilus with the method was masculine, but to Bifidobacterium bifidus,***, Lactobacillus bulgaricus, Enterococcus faecium,and Staphylococcus aureus those were existed in animal intestinal tract was negative;the lowest detect examine was 8 copies of 16S rDNA;Extracted DNA of the duodenal, jejunal, ileac, cecal ,and rectal contents of ducks at 20,21, 22,23,26,32,41 ,and 56d of age was detected with this method , results indicated these intestinal tract spots has the bacillus to exist, the quantity of Bacillus in different intestinal tract spot has not obvious difference. This method has high sensitivity and specificity, which could detect fast the quantity of Bacillus in the duck intestinal tract.

关键词： Bacillus 16S rDNA Real-time fluorescent quantitate PCR

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The dynamics of Escherichia/Staphyloccocus/Lactobacillus in Respiratory tract of ducks infected by DPV virulent strain

The dynamics of Escherichia/Staphyloccocus/Lactobacillus in ...

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第三届第八次全国学术研讨会暨动物微生态企业发展战略论坛

作者： HU Ji, CHENG An-chun, WANG Ming-shu, LIU Yan-li, CHEN Xiao-yue (Avian disease Research Center, animal Science & Technology College, sichuan Agriculture University, yaan, sichuan, 625014 the key laboratory of animal disease and human health of sichuan province, yaan, sichuan, 625014)

This experiment were based on the FQ-PCR method which were founded by our lab. I detected the dynamics of Escherichia/ Staphylococcus/ Lactobacillus genus in respiratory and respiratory tract of ducks infected by DPV virulent strain through nasal infection, oral infection and hypodermic infection and its relative inhabitation ducks. We got the results below: The Escherichia extremely is remarkable is lower than compares (P<0.01);The Staphylococcus is extremely remarkable is higher than compares (P<0.01) lives together the duck to be extremely remarkable is lower than compares (P<0.01);Lactobacillus quantity extremely is remarkable is higher than compares (P<0.01). Death duck:(1)Escherichia: Obviously is lower than compares (P<0.05).(2)Staphylococcus: Extremely obviously is lower than compares (P<0.01).(3)The Lactobacillus is extremely remarkable is lower than compares (P<0.01);Lives together to die the duck to be extremely remarkable is higher than compares (P<0.01).

关键词： DPV respiratory tract Escherichia Staphylococcus Lactobacillus

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Detecting StaphyLococcus by 16SrDNA Real-time fluorescence quantitative PCR and applying

Detecting StaphyLococcus by 16SrDNA Real-time fluorescence q...

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第三届第八次全国学术研讨会暨动物微生态企业发展战略论坛

作者： GE Zhong-yuan, CHENG An-chun, WANG Ming-shu, YANG Xiao-yan, QI Xue-feng, HUANG Yong-cheng, ZHANG Su-hui, HU Ji,CHEN Xiao-yue (Avian disease Research Center, College of animal Science and Veterinary Medicine of sichuan Agricultural University,625014 key laboratory Of animal disease and human health of sichuan province, yaan,sichuan,China,625014)

According to sequence of the bacteria 16SrDNA genes of StaphyLococcus genus in Genbank,the primers and the specific fluorogenic Taqman MGB probe were designed and synthesized. Staphylococcus aureus PCR product would be used as standard quantitative template to make the standard curve. To establish real-time fluorogenic quantitative PCR method for detecting the bacteria of Staphylococcus genus. The results showed that 5mmol/L Mg+2 and 0.36μmol/L MGB probe were optimal for this Real-time fluorescence quantitative PCR assay. The standard curve correlation is 0.998, linear-range is 104-1010 copies/μL .The bacteria of Staphylococcus genus could be specifically distinguished from Escherichia、Enterococcus、Bacillus and Bifidobacterium with the method. The sensitivity of this assay was 2.24 cells/μ*** method was applied and to detect Staphylococcus of of trachea、esophagus、duodenum、jejunum、ileum、caecum and rectum from 241/12d、61/3d、21d、22d、23d、26d、32d、41d、and 56 days old of the Cherry-valley duck. The results showed that the total of Staphylococcus of trachea、esophagus (logcop/whole) is 0.00-10.42 and 0.00-11.55, the total of Staphylococcus of duodenum、jejunum、 ileum、caecum and rectum (logcop/g) is 0.00-9.28、0.00-11.04、0.00-10.94、0.00-9.58、0.00-8.00. The Staphylococcus’ existent probability is more in trachea、ileum and caecum. The Staphylococcus quantity is more in trachea、jejunum、ileum and caecum. The Staphylococcus quantity show wave variety along with old day.

关键词： Staphylococcu 16SrDNA Real-timen fluorescence quantitative PCR duck

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The dynamics of Escherichia/Staphyloccocus/Lactobacillus in Respiratory tract of ducks infected by DPV virulent strain

The dynamics of Escherichia/Staphyloccocus/Lactobacillus in ...

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第三届第八次全国学术研讨会暨动物微生态企业发展战略论坛

作者： HU Ji, CHENG An-chun, WANG Ming-shu, LIU Yan-li, CHEN Xiao-yue Avian disease Research Center, animal Science & Technology College, sichuan Agriculture University, 625014 the key laboratory of animal disease and human health of sichuan province, yaan, sichuan, 625014

This experiment were based on the FQ-PCR method which were founded by our lab. I detected -the dynamics of Escherichia/ Staphylococcus/ Lactobacillus genus in respiratory and respiratory tract of ducks infected by DPV virulent strain through nasal infection, oral infection and hypodermic infection and its relative inhabitation ducks. We got the results below:The Escherichia extremely is remarkable is lower than compares (P<0.01);The Staphylococcus is extremely remarkable is higher than compares (P<0.01) lives together the duck to be extremely remarkable is lower than compares (P<0.01);Lactobacillus quantity extremely is remarkable is higher than compares (P<0.01). Death duck:(1)Escherichia: Obviously is lower than compares (P<0.05).(2)Staphylococcus: Extremely obviously is lower than compares (P<0.01).(3)The Lactobacillus is extremely remarkable is lower than compares (P<0.01);Lives together to die the duck to be extremely remarkable is higher than compares (P<0.01).

关键词： DPV respiratory tract Escherichia Staphylococcus Lactobacillus

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The dynamics of Escherichia/Staphyloccocus/Lactobacillus in Alimentary tract of ducks orally infected by DPV virulent strain

The dynamics of Escherichia/Staphyloccocus/Lactobacillus in ...

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第三届第八次全国学术研讨会暨动物微生态企业发展战略论坛

This experiment were based on the FQ-PCR method which were founded by our lab. I detected the dynamics of Escherichia/ Staphylococcus/ Lactobacillus genus in respiratory and alimentary tract of ducks infected by DPV virulent strain through nasal infection, oral infection and hypodermic infection and its relative inhabitation ducks. We got the results below:Esophagus: The Escherichia is extremely remarkable is higher than compares (P<0.01), also ihabitation group to be higher than compares (P<0.05);The Staphylococcus extremely is remarkable is higher than compares (P<0.01);Ihabitation duck Lactobacillus to be extremely remarkable is higher than compares (P<0.01). Duodenum: The Escherichia obviously is lower than compares (P<0.05);The Staphylococcus extremely is remarkable is lower than compares (P<0.01);The Lactobacillus extremely is remarkable is higher than compares (P<0.01). Jejunum: Ihabitation duck Escherichia obviously to be lower than compares (P<0.05);The Staphylococcus is extremely remarkable is higher than compares (P<0.01), but ihabitation duck to be extremely remarkable is lower than compares (P<0.01);The Lactobacillus extremely is remarkable is lower than compares (P<0.01). Ileum: Ihabitation duck Escherichia to be extremely remarkable is higher than compares (P<0.01);The staphylococcus extremely is remarkable is higher than compares (P<0.01);The Lactobacillus obviously is lower than compares (P<0.05), but ihabitation duck to be extremely remarkable is higher than compares (P<0.01). Cecum: The Escherichia is extremely remarkable is higher than compares (P<0.01);The staphylococcus is extremely remarkable is higher than compares (P<0.01), but ihabitation duck to be extremely remarkable is lower than compares (P<0.01);The Lactobacillus is extremely remarkable is higher than compares (P<0.01). Rectum: The Escherichia is extremely remarkable is higher than compares (P<0.01);The staphylococcus obviously is higher than compares (P<0.05);The Lactobacillus is ext

关键词： DPV alimentary tract Escherichia Staphylococcus Lactobacillus

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The dynamics of Escherichia/Staphyloccocus/Lactobacillus in Alimentary tract of ducks nasally infected by DPV virulent strain

The dynamics of Escherichia/Staphyloccocus/Lactobacillus in ...

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第三届第八次全国学术研讨会暨动物微生态企业发展战略论坛

作者： HU Ji, CHENG An-chun, WANG Ming-shu, LIU Yan-li, CHEN Xiao-yue Avian disease Research Center, animal Science & Technology College, sichuan Agriculture University, the key laboratory of animal disease and human health of sichuan province, yaan, sichuan, 625014

This experiment were based on the FQ-PCR method which were founded by our lab. I detected the dynamics of Escherichia/ Staphylococcus/ Lactobacillus genus in digestive and alimentary tract of ducks infected by DPV virulent strain through nasal infection, oral infection and hypodermic infection and its relative inhabitation ducks. The results show: Esophagus: The Escherichia is extremely remarkable is higher than compares (P<0.01);The Staphylococcus obviously is higher than compares (P<0.05);The Lactobacillus is extremely remarkable is higher than compares (P<0.01), also Inhabitation duck obviously to be higher than compares (P<0.05). Duodenum: The Escherichia obviously is lower than compares (P<0.05) Inhabitation duck to be extremely remarkable is higher than compares (P<0.01);Inhabitation duck Staphylococcus obviously to be higher than compares (P<0.05);The Lactobacillus extremely is remarkable is lower than compares (P<0.01). Jejunum: The Escherichia extremely is remarkable is higher than compares (P<0.01);The Lactobacillus is extremely remarkable is lower than compares (P<0.01). Ileum: Inhabitation duck Escherichia to be extremely remarkable is higher than compares (P<0.01);The Staphylococcus is extremely remarkable is lower than compares (P<0.01), but Inhabitation duck to be extremely remarkable is higher than compares (P<0.01);Lactobacillus quantity extremely is remarkable is higher than compares (P<0.01). Caecum: The Escherichia extremely is remarkable is higher than compares (P<0.01);The Staphylococcus is extremely remarkable is higher than compares (P<0.01), Inhabitation duck obviously to be higher than compares (P<0.05);The Lactobacillus is extremely remarkable is lower than compares (P<0.01). Rectum::The Escherichia is extremely remarkable is lower than compares (P<0.01);The staphylococcus is extremely remarkable is higher than compares (P<0.01);The Lactobacillus is extremely remarkable is higher than compares (P<0.01), but Inhabitation duck obviously to b

关键词： DPV alimentary tract Escherichia Staphylococcus Lactobacillus

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Dynamic Changes of Apoptosis in DEF Cells induced by New Type Gosling Viral Enteritis Virus

Dynamic Changes of Apoptosis in DEF Cells induced by New Typ...

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第四届第九次全国学术研讨会暨饲料和动物源食品安全战略论坛

作者： Chen Shun Cheng Anchun Wang Mingshu Chen Xiaoyue (1.Avian disease Research Center,College of animal Science and Veterinary Medicine of sichuan Agricultural University, 2.key laboratory of animal disease and human health of sichuan province,yaan,sichuan,625014,China 3.Research Center of animal Microecological Engineering,sichuan Agricultural University,yaan,sichuan,625014,P.R.China)

The monolayer Duck Embryo Fibroblast (DEF) cells were experimentally infected with New Type Gosling Viral Enteritis Virus (NGVEV) and the dynamic changes of apoptosis were detected at different time points after NGVEV infection by using transmission electron microscopy (TEM),DNA agarose gel electrophoresis and Annexin V-FITC/PI stained Fluorescence-Activated Cell Sorter (FACS).The result shows NGVEV can induce infected cells undergo apoptosis and change regularly.A series of characteristic apoptotic morphological changes including shrink of the cells,chromatin condensation and margination,as well as formation of apoptotic bodies were observed by *** typical ladder pattern of DNA fragmentation was demonstrated by agarose gel *** using flow cytometry analysis of Annexin V-FITC/PI staining,the dead,viable,apoptotic and necrotic cells could be analysis quantitatively.

关键词： New Type Gosling Viral Enteritis Virus (NGVEV) Duck Embryo Fibroblasts(DEF) Apoptosis

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Comparative Study on the Intestinal microbial community Between healthy and Orally Infected Ducklings With Salmonella Enteritidis

Comparative Study on the Intestinal microbial community Betw...

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第四届第九次全国学术研讨会暨饲料和动物源食品安全战略论坛

作者： Sheng-yan CAO An-chun CHENG Ming-shu WANG Xue-feng QI Xiao-yan YANG Nian-chun YIN Deng-chun ZHOU De-kang ZHU Qi-hui LUO Xiao-yue CHEN (Research Center of animal Microecological Engineering,sichuan Agricultural University, Avian disease Research Center,College of Veterinary Medicine of sichuan Agricultural University .key laboratory of animal disease and human health of sichuan province,yaan 625014,sichuan,China)

AIM:to analyze intestinal microbial community diversity and dynamic change of control healthy ducklings and seven-day-old ducklings infected with Salmonella Enteritidis(SE) by oral administration. METHODS:Applied Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR to analyze Intestinal microbial community diversity and dynamic change including duodenum,jejunum,ileum,cecum,rectum from healthy ducklings and seven-day-old ducklings after orally infection with SE at different time points ***:The results showed that:(1)the intestinal microbial community of the control healthy ducklings was steady.(2)The ERIC-PCR band numbers of the control healthy ducklings were the least with rectum and were the most with caecum.(3)ERIC-PCR bands of orally inoculated ducklings didn’t obviously change until 24h post inoculation(p.i.).The numbers of the ERIC-PCR bands gradually decreased from 24h to 72h p.i.,and then,with the development of disease,the band numbers gradually increased until 6 days p.i.(4)The DNAstar of staple of ERIC-PCR showed that aerobe and faeultative aerobe(Escherichia coli,Shigella,Salmonella)had become preponderant bacilli in intestine of orally infected duckings with ***:The research results provided significant datas to clarify the Intestinal microbial community diversity and dynamic change of healthy ducklings and SE orally ducklings. This study will help to understand the pathogenic mechanisms of SE infection to human and other animals

关键词： ERIC-PCR Salmonella Enteritidis Ducklings Infected with Salmonella Enteritidis Intestinal Microbial Communities

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