河南部分地区乌鸡群鸭源鸡杆菌的分离鉴定及系统发育分析

doi:10.11843/j.issn.0366-6964.2018.11.017

Abstract

Abstract:

In order to understand the epidemic situation of Gallibacterium in silky and the genetic evolution characteristics of isolates, samples were collected for the isolation of Gallibacterium anatis and q-PCR identification from two silky farms in Xinyang and Nanyang, Henan province, then isolates were detected by Gallibacterium specific PCR followed by drug susceptibility test, and the PCR amplification and sequencing of the 16S rRNA, rpoB and gyrB genes. Finally, the MegAlign program was used for calculating the similarity between isolates of G. anatis, and MEGA5.0 for constructing evolutionary tree and genetic evolution analysis. Results were as follows:Of 22 silky chickens, 16 were found positive to G. anatis, the positive rate was 72.7%; the four Gallibacterium isolates were identified as G. anatis; the four isolates were all multidrug resistant, and were sensitive to ceftriaxone, cefotaxime, amikacin and tobramycin only; the homology between the isolates and the reference G. anatis species was 99.1%-99.8% (16S rRNA), 85.4%-99.6% (rpoB), and 92.7%-99.8% (gyrB), respectively. In the 16S rRNA gene evolution tree, the four isolates and the reference G. anatis were located in the same branch; the GAC193 isolate in the rpoB gene evolution tree were located in the same branch with Pasteurella multocida, not in the same branch with other G. anatis; in the gyrB gene evolution tree, GAC017 and G. genomosp.1 were in a small branch. The results showed that G. anatis were found in the silky chickens with a high detection rate, and four strains of G. anatis were isolated, which showing serious multidrug resistance. There are significant genetic differences in the rpoB and gyrB gene of some isolates of G. anatis.

Key words: silky, Gallibacterium anatis, isolation and identification, drug sensitivity test, phylogenetic analysis

CLC Number:

S852.61

HUANGFU He-ping, DU Zhen-long, DONG Qing, WANG Hong-kui, SUN Yan-ting, GUO Hong-wei, JI Xiang-bo, JIA Han-xiao, YU Meng-wei, BU Ya-ge. Isolation,Identification and Phylogenetic Analysis of Gallibacterium anatis from Silky Population in Henan Province, China[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2451-2460.

References

[1] 王川庆,陈陆,杨霞,等. 蛋鸡群卡氏杆菌感染情况的初步研究[J]. 河南农业科学, 2008, 37(3):97-100, 103.
WANG C Q, CHEN L, YANG X,et al. A premilinary study of Gallibacterium anantis infection in laying hens[J]. Journal of Henan Agricultural Sciences, 2008, 37(3):97-100, 103. (in Chinese)
[2] 郑鹿平,杨霞,陈陆,等. 45株鸡卡氏杆菌的分离鉴定[J]. 中国兽医学报, 2010, 30(3):347-351.
ZHENG L P, YANG X, CHEN L,et al. Isolation and preliminary identification of 45 Gallibacterium anatis strains[J]. Chinese Journal of Veterinary Science, 2010, 30(3):347-351. (in Chinese)
[3] 田明星,林艳,邹年莉,等. 鸡卡氏杆菌的分离与16S rRNA基因的分析鉴定[J]. 中国畜牧兽医, 2010, 37(9):57-61.
TIAN M X, LIN Y, ZOU N L,et al. Isolation of Gallibacterium and identification and analysis of 16S rRNA gene[J]. China Animal Husbandry & Veterinary Medicine, 2010, 37(9):57-61. (in Chinese)
[4] 王珊,陈陆,付仁一,等. 我国部分地区蛋鸡群鸭源鸡杆菌血清流行病学调查[J]. 中国预防兽医学报, 2011, 33(2):114-117.
WANG S, CHEN L, FU R Y,et al. Seroepidemiological survey of Gallibacterium anatis infection in layer chicken flocks in some provinces[J]. Chinese Journal of Preventive Veterinary Medicine, 2011, 33(2):114-117. (in Chinese)
[5] 李乔晶,张九州,姬郭彪,等. 河北省鸡群鸭源鸡杆菌血清流行病学调查[J]. 河南农业科学, 2012, 41(2):132-135.
LI Q J, ZHANG J Z, JI G B,et al. Seroepidemiological survey of Gallibacterium anatis infection in chicken flocks in Hebei Province[J]. Journal of Henan Agricultural Sciences, 2012, 41(2):132-135. (in Chinese)
[6] 方翟,项玉,谷长勤,等. 1株鸭源鸡杆菌的分离与鉴定[J]. 中国兽医杂志, 2014, 50(6):3-5, 8, 49.
FANG D, XIANG Y, GU C Q,et al. Isolation and identification of a Gallibacterium anatis isolated from chicks[J]. Chinese Journal of Veterinary Medicine, 2014, 50(6):3-5, 8, 49. (in Chinese)
[7] 董青,史静柯,王海花,等. 郑州地区7株鸭源鸡杆菌的MALDI-TOF MS鉴定及其生物学特性分析[J]. 河南农业科学, 2017, 46(10):143-147.
DONG Q, SHI J K, WANG H H, et al. Identification of 7 Gallibacterium anatis strains by MALDI-TOF MS in Zhengzhou area and research on partial biological characteristics[J]. Journal of Henan Agricultural Sciences, 2017, 46(10):143-147. (in Chinese)
[8] 杨晓林,陈英,余松城,等. 1株鸽源鸡杆菌复合群3的分离和鉴定[J]. 浙江大学学报:农业与生命科学版, 2014, 40(3):249-256.
YANG X L, CHEN Y, YU S C,et al. Isolation and identification of one pigeon-origin Gallibacterium genomosp. 3[J]. Journal of Zhejiang University:Agriculture & Life Sciences, 2014, 40(3):249-256. (in Chinese)
[9] EL-ADAWY H, BOCKLISCH H, NEUBAUER H, et al. Identification, differentiation and antibiotic susceptibility of Gallibacterium isolates from diseased poultry[J]. Ir Vet J, 2018, 71:5.
[10] ELBESTAWY A R, ELLAKANY H F, EL-HAMID H S A, et al. Isolation, characterization, and antibiotic sensitivity assessment of Gallibacterium anatis biovar haemolytica, from diseased Egyptian chicken flocks during the years 2013 and 2015[J]. Poultry Sci, 2018, 97(5):1519-1525.
[11] LA DUC M T, SATOMI M, AGATA N, et al. gyrB as a phylogenetic discriminator for members of the Bacillus anthracis-cereus-thuringiensis group[J]. J Microbiol Methods, 2004, 56(3):383-394.
[12] 侯晓丽,曹清毅,潘劲草,等. 霍乱弧菌和副溶血弧菌分离株的gyrB基因系统发育分析[J]. 微生物学报, 2006, 46(6):884-889.
HOU X L, CAO Q Y, PAN J C, et al. Classification and identification of Vibrio cholerae and Vibrio parahaemolyticus isolates based on gyrB gene phylogenetic analysis[J]. Acta Microbiologica Sinica, 2006, 46(6):884-889. (in Chinese)
[13] 李路茜,李振军,楼永良. GyrB基因在诺卡菌菌种鉴定中的应用研究[J]. 疾病监测, 2016, 31(6):503-506.
LI L X, LI Z J, LOU Y L. Identification of Nocardia species based on gyrB gene[J]. Disease Surveillance, 2016, 31(6):503-506. (in Chinese)
[14] 邵高祥,束长龙,耿丽丽,等. 茶卡盐湖中耐盐抗真菌芽孢杆菌的分离和鉴定[J]. 中国生物防治学报, 2016, 32(4):440-448.
SHAO G X, SHU C L, GENG L L, et al. Isolation and identification of antifungal Bacillus with Salt-tolerant in Caka Salt Lake[J]. Chinese Journal of Biological Control, 2016, 32(4):440-448. (in Chinese)
[15] 倪萍,李美芽,葛尔宁,等. 肠道病原菌gyrB基因系统发育分析[J]. 浙江中医药大学学报, 2015, 39(7):548-554.
NI P, LI M Y, GE E N, et al. Classification and identification of enteric pathogens based on gyrB gene phylogenetic analysis[J]. Journal of Zhejiang Chinese Medical University, 2015, 39(7):548-554. (in Chinese)
[16] 何艳霞,向诗非,李浇,等. 基于16S rRNA和rpoB基因的分子系统发育分析在铜绿假单胞菌鉴定中的应用[J]. 微生物学杂志, 2016, 36(4):27-35.
HE Y X, XIANG S F, LI J, et al. Molecular phylogeny analysis based on 16S rRNA and rpoB sequences for identification of clinical isolates of Pseudomonas aeruginosa[J]. Journal of Microbiology, 2016, 36(4):27-35. (in Chinese)
[17] 皇甫和平,杨霞,赵军,等. 豫陕两省14株鸡杆菌的gyrB、16S rRNA和rpoB基因序列比较分析[J]. 畜牧兽医学报, 2012, 43(7):1103-1110.
HUANGFU H P, YANG X, ZHAO J,et al. Sequence analysis of fourteen Gallibacterium isolates from Henan and Shanxi Provinces based on gyrB, 16S rRNA and rpoB genes[J]. Acta Veterinaria et Zootechnica Sinica, 2012, 43(7):1103-1110. (in Chinese)
[18] WEISBURG W G, BARNS S M, PELLETIER D A, et al. 16S ribosomal DNA amplification for phylogenetic study[J]. J Bacteriol, 1991, 173(2):697-703.
[19] CHRISTENSEN H, KUHNERT P, OLSEN J E, et al. Comparative phylogenies of the housekeeping genes atpD, infB and rpoB and the 16S rRNA gene within the Pasteurellaceae[J]. Int J Syst Evol Microbiol, 2004, 54(5):1601-1609.
[20] JOHNSON T J, DANZEISEN J L, TRAMPEL D, et al. Genome analysis and phylogenetic relatedness of Gallibacterium anatis strains from poultry[J]. PLoS One, 2013, 8(1):e54844.
[21] HUANGFU H P, XU W B, WANG H K, et al. Detection of Gallibacterium anatis by TaqMan fluorescent quantitative PCR[J]. Avian Pathol, 2018, 47(3):245-252.
[22] BOJESEN A M, VAZQUEZ M E, ROBLES F, et al. Specific identification of Gallibacterium by a PCR using primers targeting the 16S rRNA and 23S rRNA genes[J]. Vet Microbiol, 2007, 123(1-3):262-268.
[23] KRISTENSEN B M, FREES D,BOJESEN A M. GtxA from Gallibacterium anatis, a cytolytic RTX-toxin with a novel domain organisation[J]. Vet Res, 2010, 41(3):25.
[24] 张秀平,卢彩景,陈陆,等. 鸭源鸡杆菌对鸡原代输卵管上皮细胞作用[J]. 中国兽医学报, 2016, 36(2):228-233.
ZHANG X P, LU C J, CHEN L,et al. Impact of Gallibacterium anatis on chicken oviduct epithelial cells[J]. Chinese Journal of Veterinary Science, 2016, 36(2):228-233. (in Chinese)
[25] BAGER R J, PERSSON G, NESTA B, et al. Outer membrane vesicles reflect environmental cues in Gallibacterium anatis[J]. Vet Microbiol, 2013, 167(3-4):565-572.
[26] BAGER R J, NESTA B, PORS S E, et al. The fimbrial protein FlfA from Gallibacterium anatis is a virulence factor and vaccine candidate[J]. Infect Immun, 2013, 81(6):1964-1973.
[27] GARCÍA-GÉMEZ E, VACA S, PÉREZ-MÉNDEZ A, et al. Gallibacterium anatis-secreted metalloproteases degrade chicken IgG[J]. Avian Pathol, 2005, 34(5):426-429.
[28] 王继洋,王艳,彭志锋,等. 鸭源鸡杆菌相关致病因子研究进展[J]. 中国兽医学报, 2017, 37(8):1627-1632.
WANG J Y, WANG Y, PENG Z F, et al. Research process on the virulence factors of Gallibacterium anatis in poultry[J]. Chinese Journal of Veterinary Science, 2017, 37(8):1627-1632. (in Chinese)
[29] DRANCOURT M, BOLLET C, CARLIOZ A, et al. 16S ribosomal DNA sequence analysis of a large collection of environmental and clinical unidentifiable bacterial isolates[J]. J Clin Microbiol, 2000, 38(10):3623-3630.
[30] PEI A Y, OBERDORF W E, NOSSA C W, et al. Diversity of 16S rRNA genes within individual prokaryotic genomes[J]. Appl Environ Microbiol, 2010, 76(12):3886-3897.
[31] KIM B J, KIM C J, CHUN J, et al. Phylogenetic analysis of the genera Streptomyces and Kitasatospora based on partial RNA polymerase β-subunit gene (rpoB) sequences[J]. Int J Syst Evol Microbiol, 2004, 54(2):593-598.
[32] CHRISTENSEN H, BISGAARD M, BOJESEN A M, et al. Genetic relationships among avian isolates classified as Pasteurella haemolytica. ‘Actinobacillus salpingitidis’ or Pasteurella anatis with proposal of Gallibacterium anatis gen. nov., comb. Nov. and description of additional genomospecies within Gallibacterium gen. nov.[J]. Int J Syst Evol Microbiol, 2003, 53(Pt 1):275-287.
[33] BISGAARD M, KORCZA B M, BUSSE H J, et al. Classification of the taxon 2 and taxon 3 complex of bisgaard within Gallibacterium and description of Gallibacterium melopsittaci sp. nov., Gallibacterium trehalosifermentans sp. nov. and Gallibacterium salpingitidis sp. nov.[J]. Int J Syst Evol Microbiol, 2009, 59(4):735-744.
[34] 郭伦涛. 鸡杆菌地方分离株耐药性及耐药基因的研究[D]. 郑州:河南农业大学, 2010.
GUO L T. Studies on drug resistance and resistant genes of Gallibacterium anatis strains isolated from chickens in different localities[D]. Zhengzhou:Henan Agricultural University, 2010. (in Chinese)
[35] 高冬生. 61株鸭源鸡杆菌部分耐药基因及其与耐药性关系的研究[D]. 郑州:河南农业大学, 2011.
GAO D S.The correlation between drug resistance and some drug resistance genes in sixty-one Gallibacterium anatis isolates[D]. Zhengzhou:Henan Agricultural University, 2011. (in Chinese)
[36] 彭志锋,高冬生,刘红英,等. 鸭源鸡杆菌整合子及其与耐药性的相关性分析[J]. 畜牧兽医学报, 2016, 47(8):1676-1681.
PENG Z F, GAO D S, LIU H Y, et al. Gallibacterium anatis Integron and its correlation with the drug resistance[J]. Acta Veterinaria et Zootechnica Sinica, 2016, 47(8):1676-1681. (in Chinese)

Isolation,Identification and Phylogenetic Analysis of Gallibacterium anatis from Silky Population in Henan Province, China

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LIU Qi, SHI Lijun, LIANG Lin, ZHANG Lingling, YUAN Weifeng, LIANG Ruiying, LI Jinxiang, CUI Shangjin. Isolation and Identification of a Feline Panleukopenia Virus and Genetic Variation Analysis of Its VP2 and NS1 [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 1106-1112.
[2]	LIU Qi, SHI Lijun, LIANG Lin, ZHANG Lingling, YUAN Weifeng, LIANG Ruiying, LI Jinxiang, CUI Shangjin. Isolation and Identification of a Feline Panleukopenia Virus and Genetic Variation Analysis of Its VP2 and NS1 [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 1106-1112.
[3]	YANG Zhenxing, MENG Jinxin, XIAO Lei, ZHU Jianbo, LIAO Defang, GAO Lin, LI Zhanhong, YANG Heng, LI Huachun. Isolation of Epizootic Haemorrhagic Disease Virus Serotype 7 Strain in China for the First Time [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(3): 602-610.
[4]	YANG Zhenxing, MENG Jinxin, XIAO Lei, ZHU Jianbo, LIAO Defang, GAO Lin, LI Zhanhong, YANG Heng, LI Huachun. Isolation of Epizootic Haemorrhagic Disease Virus Serotype 7 Strain in China for the First Time [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(3): 602-610.
[5]	WEN Fengqin, XIANG Haitao, HAO Baocheng, XING Xiaoyong, BAO Shijun, HU Yonghao. Isolation and Identification of Corynebacterium Xerosis Isolated from Yak Liver [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(1): 227-232.
[6]	WEN Fengqin, XIANG Haitao, HAO Baocheng, XING Xiaoyong, BAO Shijun, HU Yonghao. Isolation and Identification of Corynebacterium Xerosis Isolated from Yak Liver [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(1): 227-232.
[7]	SUN Lu, REN Zhen-hui, SONG Run-jie, QUAN Hai-yun, JIAO Yun-jie, HUANG Zhu-tao, WANG Jing-long, ZHAO Bao-yu, LU Hao. Isolation, Identification and Phylogenetic Analysis of Fungal Endophyte in Oxytropis trichophora [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 2025-2035.
[8]	SUN Lu, REN Zhen-hui, SONG Run-jie, QUAN Hai-yun, JIAO Yun-jie, HUANG Zhu-tao, WANG Jing-long, ZHAO Bao-yu, LU Hao. Isolation, Identification and Phylogenetic Analysis of Fungal Endophyte in Oxytropis trichophora [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 2025-2035.
[9]	WEI Run-yu, LUO Yao-yao, Lü Yan, WANG Jing-jing, ZHENG Dong-xia, ZHAO Yun-ling, LIU Wen-bo, LIU Hua-lei, WANG Zhi-liang. Distribution and Molecular Characterization of 4 New Class Ⅰ Newcastle Disease Viruses Isolated from China [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1781-1788.
[10]	HAO Fei, MAO Li, LI Wen-liang, LI Ji-zong, YANG Lei-lei, ZHANG Wen-wen, SUN Min, ZHONG Chun-yan, LIU Mao-jun, JIANG Jie-yuan. Identification and Pathogenicity of One Caprine Herpesvirus 1 Strain [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1797-1802.
[11]	WEI Run-yu, LUO Yao-yao, Lü Yan, WANG Jing-jing, ZHENG Dong-xia, ZHAO Yun-ling, LIU Wen-bo, LIU Hua-lei, WANG Zhi-liang. Distribution and Molecular Characterization of 4 New Class Ⅰ Newcastle Disease Viruses Isolated from China [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1781-1788.
[12]	HAO Fei, MAO Li, LI Wen-liang, LI Ji-zong, YANG Lei-lei, ZHANG Wen-wen, SUN Min, ZHONG Chun-yan, LIU Mao-jun, JIANG Jie-yuan. Identification and Pathogenicity of One Caprine Herpesvirus 1 Strain [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1797-1802.
[13]	JI Cheng-yuan, WANG Wei-yi, FENG Xuan-rong, YAO Jing, HUANG Xin, FENG Yuan-qing, QIN Yi-feng, FANG Qing-li, CHEN Ying, OUYANG Kang, WEI Zu-zhang, HUANG Wei-jian. Epidemiological Investigation and Genetic Diversity Analysis of Porcine Circovirus Type 3 in Slaughter Pigs of a City in Guangxi [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1265-1273.
[14]	JI Cheng-yuan, WANG Wei-yi, FENG Xuan-rong, YAO Jing, HUANG Xin, FENG Yuan-qing, QIN Yi-feng, FANG Qing-li, CHEN Ying, OUYANG Kang, WEI Zu-zhang, HUANG Wei-jian. Epidemiological Investigation and Genetic Diversity Analysis of Porcine Circovirus Type 3 in Slaughter Pigs of a City in Guangxi [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1265-1273.
[15]	CHEN Yao, DING Hong-lei, HE Ying, MU Hao, WANG Li-min, WANG Hao-ju. Antimicrobial Susceptibility Analysis and ESBL Gene Detection of Staphylococcus aureus Isolated at Animal Farms in Chongqing [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 1074-1080.