竹鼠源弗氏柠檬酸杆菌的分离鉴定及致病性分析

doi:10.11843/j.issn.0366-6964.2017.05.017

摘要/Abstract

摘要：

为确定桂林某竹鼠养殖场患病竹鼠的病原，从发病竹鼠肝组织分离到一株细菌。根据形态学特征及16S rRNA基因序列分析，确定分离菌株为弗氏柠檬酸杆菌。进化分析显示，该菌与来自棉花红蜘蛛的弗氏柠檬酸杆菌遗传关系最近，但处于独立的分支上。毒力试验结果表明，该菌对成年小白鼠的LD₅₀为2.8×10⁸CFU，对10~20日龄的竹鼠LD₅₀为6.3×10⁸ CFU。药敏试验结果显示，分离菌株对头孢曲松、庆大霉素、阿米卡星、氧氟沙星、左氧氟沙星药物高度敏感，对青霉素、头孢唑啉、氟苯尼考、林可霉素、强力霉素等有耐药性。病理组织学观察显示，发病竹鼠肺泡间隔增宽伴随炎性细胞浸润；肝组织血管扩张、淤血；脾被膜外大量坏死细胞；肾组织肾小管上皮细胞肿胀、颗粒变性。综上研究结果表明，弗氏柠檬酸杆菌是引起桂林竹鼠发病和致死的病原菌，分离菌株对竹鼠有较强的致病性。

Abstract:

To identify the causative agent infecting the bamboo rats on a farm in the Guilin, a strain of bacterium was isolated from the liver of diseased bamboo rats. Its morphological characteristics and the results of 16S rRNA gene sequence analysis proved that the pathogenic bacteria was Citrobacter freundii. Phylogenetic analysis indicated that the isolate was closely related to C. freundii from a two-spotted spider mite, but it was an independent branch. In a pathogenic test, the isolate was pathogenic to adult mice, with a median lethal dose (LD₅₀) of 2.8 ×10⁸ CFU, and its LD₅₀to 10-20 days old bamboo rats was 6.3×10⁸ CFU. Results of antibiotic susceptibility testing demonstrated that the C. freundii was highly sensitive to ceftriaxone, gentamycin, amikacin, ofloxacin, levofloxacin, and was resistant to penicillin, cefazolin, florfenicol, lincomycin, deoxycycline. Histopathological observation showed that the bamboo rat's septum alveolus widened, accompanied by inflammatory cell infiltration, and angiectasis in liver and hyperemia. Outside the integument of spleen there were a large number of necrotic cells, and the swelling and granular degeneration of the renal tubular epithelial cells. These findings suggest that C. freundii is the pathogen of the dead bamboo rats in Guilin. The pathogenicity of the isolate to bamboo rats is strong.

中图分类号:

S852.61

唐海波, 陈凤莲, 饶桂波, 姜佳佳, 孟菲, 白安斌, 吴健敏. 竹鼠源弗氏柠檬酸杆菌的分离鉴定及致病性分析[J]. 畜牧兽医学报, 2017, 48(5): 922-929.

TANG Hai-bo, CHEN Feng-lian, RAO Gui-bo, JIANG Jia-jia, MENG Fei, BAI An-bin, WU Jian-min. Isolation and Identification of Citrobacter freundii from the Diseased Bamboo Rats and Analysis of the Pathogenicity of the Isolate[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(5): 922-929.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2017.05.017

https://www.xmsyxb.com/CN/Y2017/V48/I5/922

参考文献

[1] 曾元根, 邓治邦. 中华竹鼠大肠杆菌病诊断与防制[J]. 动物医学进展, 2002, 23(3): 94-95.
ZENG Y G, DENG Z B. Studies on isolation and identification of Escherichia coli in Bamboo Rat[J]. Progress in Veterinary Medicine, 2002, 23(3): 94-95. (in Chinese)
[2] 刘欢, 苏丹萍, 陈敏鸿, 等. 竹鼠巴氏杆菌病的诊治[J]. 中国畜牧兽医, 2009, 36(10): 165-168.
LIU H, SU D P, CHEN M H, et al. Diagnosis and treatment of pasteurellosis in Rhizomys sinensis[J]. China Animal Husbandry & Veterinary Medicine, 2009, 36(10): 165-168. (in Chinese)
[3] 祝宇, 时永强, 王奇惠, 等. 竹鼠沙门氏菌与链球菌混合感染的诊断[J]. 中兽医学杂志, 2016(4): 49-51.
ZHU Y, SHI Y Q, WANG Q H, et al. Diagnosis of coinfection with Salmonella and Streptococcus in Rhizomys sinensis[J]. Chinese Journal of Traditional Veterinary Science, 2016(4): 49-51. (in Chinese)
[4] 马磊, 颜其贵, 万莉, 等. 竹鼠肺炎克雷伯氏菌的分离鉴定[J]. 中国人兽共患病学报, 2011, 27(9): 825-827.
MA L, YAN Q G, WAN L, et al. Isolation and identification on K. peneumoniae in Rhizomys[J]. Chinese Journal of Zoonoses, 2011, 27(9): 825-827. (in Chinese)
[5] 张宝元. 10%福尔马林与中性福尔马林的比较[J]. 四川解剖学杂志, 2001, 9(2): 97.
ZHANG B Y. The comparison of 10% formaldehyde and neutral formaldehyde[J]. Sichuan Journal of Anatomy, 2001, 9(2): 97. (in Chinese)
[6] 谢彬, 袁洁, 袁书智, 等. 猪源绿色气球菌的分离鉴定与药敏特性研究[J]. 中国微生态学杂志, 2010, 22(3): 238-243.
XIE B, YUAN J, YUAN S Z, et al. Isolation and identification of Aerococcus viridans from swine and research on its susceptibility characteristics[J]. Chinese Journal of Microecology, 2010, 22(3): 238-243. (in Chinese)
[7] TAMURA K, PETERSON D, PETERSON N, et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Mol Biol Evol, 2011, 28(10): 2731-2739.
[8] 王道若. 微生物学[M]. 3版. 北京: 人民卫生出版社, 1994: 164.
WANG D R. Microbiology[M]. 3rd ed. Beijing: People's Medical Publishing House, 1994: 164. (in Chinese)
[9] 薛巧, 赵战勤, 刘会胜, 等. 弗氏柠檬酸杆菌对动物和人致病性研究进展[J]. 动物医学进展, 2015, 36(7): 81-85.
XUE Q, ZHAO Z Q, LIU H S, et al. Progress on Citrobacter freundii pathogenicity in animals and humans[J]. Progress in Veterinary Medicine, 2015, 36(7): 81-85. (in Chinese)
[10] 纪新梅, 赵奋勤, 张云梅, 等. 小儿枸橼酸杆菌性肠炎110例临床分析[J]. 中华医院感染杂志, 1997, 7(2): 95-96.
JI X M, ZHAO F Q, ZHANG Y M, et al. Clinical analysis of 110 citrobacter enteritis children[J]. Chinese Journal of Nosocomiology, 1997, 7(2): 95-96. (in Chinese)
[11] 孙晓春. 新生儿弗劳第枸橼酸杆菌脑膜炎并阻塞性脑积水1例[J]. 中国现代医学杂志, 1997, 7(1): 46.
SUN X C. Citrobacter freundii caused meningitis and obstructive hydrocephalus in newborn[J]. China Journal of Modern Medicine, 1997, 7(1): 46. (in Chinese)
[12] 余德文, 田梅兰, 王根春. 弗劳地枸橼酸杆菌败血症11例报告[J]. 西南国防医药, 1992, 2(6): 348-349.
YU D W, TIAN M L, WANG G C. The report of 11 citrobacter freundii caused blood poisoning in human[J]. Medical Journal of National Defending Forces in Southwest China, 1992, 2(6): 348-349. (in Chinese)
[13] 胡秀彩, 王艺, 吕爱军. 弗氏柠檬酸杆菌的分离鉴定与PCR-SSCP分析[J]. 微生物学杂志, 2011, 31(4): 12-18.
HU X C, WANG Y, LV A J. Isolation and identification of Citrobacter freundii and its PCR-SSCP analysis[J]. Journal of Microbiology, 2011, 31(4): 12-18. (in Chinese)
[14] 王利, 魏勇. 鲫鱼弗氏柠檬酸杆菌的鉴定及系统发育分析[J]. 水产科学, 2012, 31(8): 481-484.
WANG L, WEI Y. Identification and phylogenetic analysis of bacterium Citrobacter freundii from crucian carp Carassius auratus by 16S rDNA genes[J]. Fisheries Science, 2012, 31(8): 481-484. (in Chinese)
[15] 吕爱军, 胡秀彩, 朱静榕, 等. 弗氏柠檬酸杆菌感染诱导斑马鱼皮肤免疫相关基因的差异表达[J]. 水产学报, 2012, 36(3): 359-366.
LV A J, HU X C, ZHU J R, et al. Differential expression of immune-related genes from skin of zebrafish induced by the infection of Citrobacter freundii[J]. Journal of Fisheries of China, 2012, 36(3): 359-366. (in Chinese)
[16] 杨移斌, 夏永涛, 赵蕾, 等. 鲟源弗氏柠檬酸杆菌分离鉴定及药敏特性研究[J]. 水生生物学报, 2013, 37(4): 766-771.
YANG Y B, XIA Y T, ZHAO L, et al. Isolation and identification of Citrobacter freundii from amur sturgeon (Acipenser schrenckii brandt) and its antibiotic sensitivity[J]. Acta Hydrobiologica Sinica, 2013, 37(4): 766-771. (in Chinese)
[17] 林启存, 朱丽敏, 李忠全, 等. 中华鳖弗氏柠檬酸杆菌败血症病原分离鉴定与药敏试验[J]. 水产科学, 2008, 27(1): 42-43.
LIN Q C, ZHU L M, LI Z Q, et al. Isolation, determination and antimicrobial susceptibility test of the Citrobacter freundii septicemia from soft shelled turtle Trionyx sinensis[J]. Fisheries Science, 2008, 27(1): 42-43. (in Chinese)
[18] 沈锦玉, 顾志敏, 潘晓艺, 等. 红螯螯虾弗氏柠檬酸杆菌病病原的分离与鉴定[J]. 中国水产科学, 2005, 12(2): 197-200.
SHEN J Y, GU Z M, PAN X Y, et al. Isolation and identification of Citrobacter freundii from Cherax quadricarinatus[J]. Journal of Fishery Sciences of China, 2005, 12(2): 197-200. (in Chinese)
[19] 雷静, 练耕宁, 牟平, 等. 从犊牛腹泻病料中分离到两株弗劳地枸橼酸盐杆菌的报告[J]. 中国奶牛, 2007(7): 33-34.
LEI J, LIAN G N, MOU P, et al. The report of isolation of 2 Citrobacter freundii from calf diarrhea[J]. China Dairy Cattle, 2007(7): 33-34. (in Chinese)
[20] 孙洋, 郭学军, 周博, 等. 东北虎出血性腹泻病原分离与鉴定[J]. 中国兽医杂志, 2008, 44(9): 89-90.
SUN Y, GUO X J, ZHOU B, et al. Isolation and identification of the pathogen caused bloody diarrhea in Siberian tiger[J]. Chinese Journal of Veterinary Medicine, 2008, 44(9): 89-90. (in Chinese)
[21] 刘会胜, 赵战勤, 薛云, 等. 羊源致病性弗氏柠檬酸杆菌的分离与鉴定[J]. 畜牧兽医学报, 2015, 46(9): 1593-1599.
LIU H S, ZHAO Z Q, XUE Y, et al. Isolation and identification of Citrobacter freundii from the diseased small tail Han sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2015, 46(9): 1593-1599. (in Chinese)