鸡白痢沙门菌减毒活疫苗候选株SKLZΔspiC对非靶动物(鼠)的致病性
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摘要
目的评价鸡白痢沙门菌减毒活疫苗候选株SKLZΔspiC对非靶动物(鼠)的致病性。方法采用鸡白痢沙门菌减毒活疫苗候选株SKLZΔspi C[1×10~7、1×10~9 CFU/(0. 1 mL·只)]及鸡白痢沙门菌S06004(Nal~R)[1×10~7 CFU/(0. 1 mL·只)],分别经肌肉注射和口服感染小鼠,同时设空白对照组(未感染)。观察小鼠的临床症状、病理、体重、脏器内菌体载量、粪便排菌及脾脏细胞因子水平,分析鸡白痢沙门菌减毒疫苗候选株对小鼠的致病性。结果与空白对照组比较,各组小鼠均未出现任何临床症状,肝脏、脾脏、盲肠病理观察均未见任何病变,体重变化差异无统计学意义(P> 0. 05)。与鸡白痢沙门菌S06004(Nal R)感染组比较,减毒活疫苗候选株感染组小鼠肝脏、脾脏、盲肠内及小鼠粪便载菌量均降低。各感染组小鼠脾脏中炎性因子IL-1β、IFNγ、IL-12、IL-6表达量均较低,仅引起小鼠轻微短暂的炎症反应。结论鸡白痢沙门菌减毒疫苗候选株SKLZΔspiC对非靶动物(鼠)无致病性,符合公共卫生安全要求。
Objective To evaluate the pathogenicity of live attenuated Salmonella pullorum vaccine candidate strain SKLZΔspiC to non-target animals(mice). Methods Mice were infected with S. pullorum SKLZΔspiC strain at dosages of 1 × 10~7 and 1 × 10~9 CFU/0. 1 mL and its parent strain S06004(Nal~R)at a dosage of 1 × 10~7 CFU/0. 1 mL by intramuscular and oral routes separately,using those uninfected as control. The clinical symptoms,pathological changes,bodyweights,bacterial loads in liver,speen and cecal,fecal excretion and splenic inflammatory cytokines of mice were observed to analyze the pathogenicity of SKLZΔspiC strain to mice. Results Compared with those in blank control group,no any clinical symptoms were observed in mice in various test groups,and no any lesions in the histopathology of liver,speen and cecal,while the bodyweight showed no significant change(P > 0. 05). However,compared with those infected with strain S06004(Nal R),the bacterial loads in liver,speen and cecal as well as fecal excretion of mice infected with SKLZΔspiC strain decreased. The expression levels of IL-1β,IFNγ,IL-12 and IL-6 in various test groups were low,which caused only mild and transient inflammatory reactions. Conclusion Live attenuated S. pullorum vaccine candidate strain SKLZΔspiC showed no pathogenicity to non-target animals(mice),which met the requirements of public safety.
Objective To evaluate the pathogenicity of live attenuated Salmonella pullorum vaccine candidate strain SKLZΔspiC to non-target animals(mice). Methods Mice were infected with S. pullorum SKLZΔspiC strain at dosages of 1 × 10~7 and 1 × 10~9 CFU/0. 1 mL and its parent strain S06004(Nal~R)at a dosage of 1 × 10~7 CFU/0. 1 mL by intramuscular and oral routes separately,using those uninfected as control. The clinical symptoms,pathological changes,bodyweights,bacterial loads in liver,speen and cecal,fecal excretion and splenic inflammatory cytokines of mice were observed to analyze the pathogenicity of SKLZΔspiC strain to mice. Results Compared with those in blank control group,no any clinical symptoms were observed in mice in various test groups,and no any lesions in the histopathology of liver,speen and cecal,while the bodyweight showed no significant change(P > 0. 05). However,compared with those infected with strain S06004(Nal R),the bacterial loads in liver,speen and cecal as well as fecal excretion of mice infected with SKLZΔspiC strain decreased. The expression levels of IL-1β,IFNγ,IL-12 and IL-6 in various test groups were low,which caused only mild and transient inflammatory reactions. Conclusion Live attenuated S. pullorum vaccine candidate strain SKLZΔspiC showed no pathogenicity to non-target animals(mice),which met the requirements of public safety.
引文
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[19]DUTTA S,DAS S,MITRA U,et al.Antimicrobial resistance,virulence profiles and molecular subtypes of Salmonella enterica serovars typhi and paratyphi A blood isolates from Kolkata,India during 2009-2013[J].Plos One,2014,9(8):e101347.
[20]GENG S,JIAO X,BARROW P,et al.Virulence determinants of Salmonella gallinarum biovar pullorum identified by PCRsignature-tagged mutagenesis and the spiC mutant as a candidate live attenuated vaccine[J].Vet Microbiol,2014,168(2-4):388-394.
[21]KILROY S,RASPOET R,HAESEBROUCK F,et al.Prevention of egg contamination by Salmonella enteritidis after oral vaccination of laying hens with Salmonella enteritidisΔtolC andΔacrABacrEFmdtABC mutants[J].Vet Res,2016,47(1):82.
[22]AN J H,Infection and cytokine storm[J].Chin J Exp Clin Infect Dis(Electronic Ed),2013,7(6):925-926.(in Chinese)安纪红.感染与细胞因子风暴[J].中华实验和临床感染病杂志(电子版),2013,7(6):925-926.
[2]ZHU L Y,ZHENG L L,BI J L,et al.Isolation,identification and drug resistance analysis of bacterial pathogen of piglets diarrhea syndrome in Qujing Area[J].Chin Animal Husbandry&Veter Med,2015,42(3):696-700.(in Chinese)朱玲云,郑龙龙,毕峻龙,等.曲靖地区仔猪腹泻细菌性病原的分离鉴定与耐药性分析[J].中国畜牧兽医,2015,42(3):696-700.
[3]BARROW P A,METHNER U.Salmonella in domestic animals[M].Wallingford:CABI Publishing,2013.
[4]GUO R,GENG S,JIAO H,et al.Evaluation of protective efficacy of a novel inactivated Salmonella pullorum ghost vaccine against virulent challenge in chickens[J].Vet Immunol Immuno Pathol,2016,173:27-33.
[5]CHENG Z,YIN J,KANG X,et al.Safety and protective efficacy of a spiC and crp deletion mutant of Salmonella gallinarum as a live attenuated vaccine for fowl typhoid[J].Res Vet Sci,2016,107:50-54.
[6]GONG J,ZHANG J,XU M,et al.Prevalence and fimbrial genotyPe distribution of poultry Salmonella isolates in China(2006 to 2012)[J].Appl Environ Microbiol,2014,80(2):687-693.
[7]LI X.The characteristics of antimicrobial resistance and related genes of Salmonella pullorum isolates in parts of China[D].Yangzhou:Yangzhou University,2016.(in Chinese)李秀.我国部分地区鸡白痢沙门菌耐药性与耐药基因分析[D].江苏扬州:扬州大学,2016.
[8]DESIN T S,KORSTER W,POTTER A A.Salmonella vaccines in poultry:past,present and future[J].Expert Rev Vaccines,2013,12(1):87-96.
[9]OKAMURA M,TACHIZAKI H,KUBO T,et al.Comparative evaluation of a bivalent killed Salmonella vaccine to prevent egg contamination with Salmonella enterica serovars enteritidis,typhimurium,and Gallinarum biovar pullorum,using 4 different challenge models[J].Vaccine,2007,25(25):4837-4844.
[10]WIGLEY P.Salmonella enterica serovar gallinarum:addressing fundamental questions in bacteriology sixty years on from the9R vaccine[J].Avian Pathol,2017,46(2):119-124.
[11]LIU N N.Construction of Salmonella enterica serovar pullorum S06004ΔspiC and its immunobiological features[D].Yangzhou:Yangzhou University,2012.(in Chinese)刘男男.鸡白痢沙门菌S06004ΔspiC突变株的构建及其免疫生物学特性初步研究[D].江苏扬州:扬州大学,2012.
[12]LIU Y F,YOU M S.Management and evaluating methods of biosafety of transgenic crops in China[J].Wuyi Sci J,2002(00):46-50.(in Chinese)刘雨芳,尤民生.中国农业转基因生物的安全性管理与评价方法[J].武夷科学,2002(00):46-50.
[13]ISSENHUTH-JEANJEAN S,ROGGENTIN P,MIKOLEIT M,et al.Supplement 2008-2010(no.48)to the White-KauffmannLe Minor scheme[J].Res Microbiol,2014,165(7):526-530.
[14]GENG S Z,GUO R X,JIAO X A,et al.Advances in Salmonella pullorum[J].China Poultry,2014,36(5):34-37.(in Chinese)耿士忠,郭荣显,焦新安,等.鸡白痢沙门菌研究进展[J].中国家禽,2014,36(5):34-37.
[15]GUO R,JIAO Y,LI Z,et al.Safety,protective immunity,and DIVA capability of a rough mutant Salmonella pullorum vaccine candidate in broilers[J].Front Microbiol,2017,8:547.
[16]LIU G,RAHN A,LIU W,et al.The evolving genome of Salmonella enterica serovar pullorum[J].J Bacteriol,2002,184(10):2626-2633.
[17]YIN J,CHENG Z,WANG X,et al.Evaluation of the Salmonella enterica serovar pullorum pathogenicity island 2mutant as a candidate live attenuated oral vaccine[J].Clin Vaccine Immunol,2015,22(7):706-710.
[18]FOLEY S,NAYAK R,HANNING I,et al.Population dynamics of Salmonella enterica serotypes in commercial egg and poultry production[J].Appl Environ Microbiol,2011,77(13):4273-4279.
[19]DUTTA S,DAS S,MITRA U,et al.Antimicrobial resistance,virulence profiles and molecular subtypes of Salmonella enterica serovars typhi and paratyphi A blood isolates from Kolkata,India during 2009-2013[J].Plos One,2014,9(8):e101347.
[20]GENG S,JIAO X,BARROW P,et al.Virulence determinants of Salmonella gallinarum biovar pullorum identified by PCRsignature-tagged mutagenesis and the spiC mutant as a candidate live attenuated vaccine[J].Vet Microbiol,2014,168(2-4):388-394.
[21]KILROY S,RASPOET R,HAESEBROUCK F,et al.Prevention of egg contamination by Salmonella enteritidis after oral vaccination of laying hens with Salmonella enteritidisΔtolC andΔacrABacrEFmdtABC mutants[J].Vet Res,2016,47(1):82.
[22]AN J H,Infection and cytokine storm[J].Chin J Exp Clin Infect Dis(Electronic Ed),2013,7(6):925-926.(in Chinese)安纪红.感染与细胞因子风暴[J].中华实验和临床感染病杂志(电子版),2013,7(6):925-926.