甘肃地区犊牛腹泻病原调查及产气荚膜梭菌的分离鉴定、耐药性分析

doi:10.16431/j.cnki.1671-7236.2025.08.041

Abstract

Abstract: 【Objective】 This experiment was to investigate the prevalence of infectious pathogenic microorganisms causing diarrhea in calves from large-scale cattle farms in Gansu and to understand the resistance status of Clostridium perfringens (Cp) to common antibacterial drugs. 【Method】 5-combined nucleic acid detection kit for pathogenic microorganisms of calf diarrhea was used to test 108 fecal samples of calf diarrhea collected from 10 cattle farms in 8 regions including Lanzhou,Zhangye and Wuwei，which mainly included Cp,Bovine rotavirus (BRV),Bovine coronavirus (BCoV),enterotoxigenic Escherichia coli K99 (K99) and Cryptosporidium (COWP). Cp nucleic acid positive bovine fecal samples were subject to bacterial isolation and culture,and identified using staining microscopy and biochemical tests.At the same time,PCR amplification was used to type the isolated strains.The drug sensitivity test and drug resistance gene detection of the isolated strains were conducted by disk diffusion method (K-B) and PCR，respectively. 【Result】 Among the 108 fecal samples collected from calves with diarrhea,the positive rate of Cp nucleic acid was 78.70% (85/108),the positive rate of individual infection was 14.81% (16/108),and the total mixed infection rate was 63.89% (69/108).The mixed infection rates of Cp with BRV and BCoV was 44.44% (48/108) and 24.07% (26/108),respectively.A total of 13 Cp were isolated,and all of them were type A.The results of drug sensitivity test showed that 84.62% (11/13) of the isolated strains were resistant to gentamicin,and 76.92% (10/13) were resistant to kanamycin.The results of the resistance gene detection showed that 84.62% (11/13) of the isolated strains were detected with Acc(6')/ph(2″) gene,53.85% (7/13) of the isolated strains were detected with ermC gene,and none of the seven resistance genes such as Sul1,qnrS,and ermB were detected. 【Conclusion】 The mixed infection of calf diarrhea in Gansu region was relatively serious.The main prevalent Cp strain was type A，and most Cp isolated strains were severely resistant to aminoglycoside and carried multiple resistance genes.

Key words: Gansu; calf diarrhea; Clostridium perfringens; isolation and identification; drug resistance

CLC Number:

S852.61^＋6.3

ZHAO Xuejian, CHEN Xiaoliang, FENG Xiaowei, ZHANG Kang, MA Xiaojun, LI Jianxi. Investigation of the Pathogen of Calf Diarrhea in Gansu Region and Isolation, Identification and Drug Resistance Analysis of Clostridium perfringens[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3927-3938.

References

[1] 高海慧,黎玉琼,高小斐,等.犊牛腹泻源大肠杆菌耐药情况及HPI相关基因的检测[J].中国畜牧兽医,2020,47(6):1942-1951. GAO H H,LI Y Q,GAO X F,et al.Drug resistance and detection of HPI related genes in E.coli from calves with diarrhea[J].China Animal Husbandry & Veterinary Medicine,2020,47(6):1942-1951.(in Chinese)
[2] 王玙，王建强.甘肃陇东南规模养牛场常发二三类牛病的血清流行病学调查[J].中国兽医杂志,2021,57(3):11-13. WANG Y,WANG J Q.Seroepidemiological investigation of common class Ⅱ and Ⅲ cattle diseases in the Southeastern of Tianshui local cattle farms[J].Chinese Journal of Veterinary Medicine,2021,57(3):11-13.(in Chinese)
[3] 吴克.西北地区羊源产气荚膜梭菌分离鉴定及生物特性分析[D].杨凌：西北农林科技大学,2023. WU K.Isolation and biological characteristic analysis of sheep/goat-derived Clostridium perfringens in Northwest China[D].Yangling：Northwest A&F University,2023.(in Chinese)
[4] VERMA K A,ABDEL-GLIL Y M,MADESH A,et al.Multilocus sequence typing of Clostridium perfringens strains from neonatal calves,dairy workers and associated environment in India[J].Anaerobe,2020,63:102212.
[5] 陆承平.兽医微生物学[M].北京:中国农业出版社,2012. LU C P.Veterinary Microbiology[M].Beijing:China Agriculture Press,2012.(in Chinese)
[6] XIU L,LIU Y,WU W,et al.Prevalence and multilocus sequence prevalence and multilocus sequence typing of Clostridium perfringens isolated from 4 duck farms in Shandong province,China[J].Poultry Science,2020,99(10):5105-5117.
[7] 候子栋,祖新政,翁业斌,等.规模化奶牛养殖场犊牛梭菌性肠炎的诱发因素和防制[J].草食家畜,2020,6:32-36. HOU Z D,ZU X Z,WENG Y B,et al.Predisposing factors and control of clostridial enteritis in calves in a large-scale dairy farms[J].Grass-feeding Livestock,2020,6:32-36.(in Chinese)
[8] KIU R,CAIM S,ALEXANDER S,et al.Probing genomic aspects of the multi-host pathogen Clostridium perfringens eveals significant pangenome diversity,and a diverse array of virulence factors[J].Frontiers in Microbiology，2017，8:2485.
[9] LUCEY B P，HUTCHINS G M.William H.Welch,MD,and the discovery of Bacillus welchii[J].Archives of Pathology & Laboratory Medicine,2004,128(10):1193-1195.
[10] BRYNESTAD S,GRANUM P E.Clostridium perfringens and foodborne infections[J].International Journal of Food Microbiology,2002,74(3):195-202.
[11] YAMAMOTO Y,IYOH N,SUGIYAMA T,et al.Clinical features of Clostridium bacteremia in cancer patients:A case series review[J].Journal of Infection and Chemotherapy,2020,26(1):92-94.
[12] LI J,MCCLANE B A.Further comparison of temperature effects on growth and survival of Clostridium perfringens type A isolates carrying a chromosomal or plasmid-borne enterotoxin gene[J].Applied and Environmental Microbiology，2006，72(7):4561-4568.
[13] 刘伟德.仔猪梭菌性肠炎的流行及诊治[J].畜牧兽医科技信息,2017,3:85-86. LIU W D.Prevalence and diagnosis and treatment of clostridial enteritis in piglets[J].Chinese Journal of Animal Husbandry and Veterinary Medicine,2017,3:85-86.(in Chinese)
[14] MUDASSAR M,ZAHID I,ABUBAKAR S,et al.Prevalence,genotypic and phenotypic characterization and antibiotic resistance profile of Clostridium perfringens type A and D isolated from feces of sheep (Ovis aries) and goats (Capra hircus) in Punjab,Pakistan[J].Toxins,2020,12(10):657.
[15] UZAL A F,MCCLANE A B,CHEUNG K J,et al.Animal models to study the pathogenesis of human and animal Clostridium perfringens infections[J].Veterinary Microbiology,2015,179(1-2):23-33.
[16] EUNTAE K,SANGJIN L,TAEYONG K,et al.Dynamic changes infecal microbial communities of neonatal dairy calves by aging and diarrhea[J].Animals,2021,11(4):1113.
[17] FINNIE J W,UZAL F A.Pathology and pathogenesis of brain lesions produced by Clostridium perfringens type D epsilon toxin[J].International Journal of Molecular Sciences,2022,23(16):9050.
[18] 董洁,杨晓静,苗承霞,等.产气荚膜梭菌菌落多重PCR方法的建立及初步应用[J].中国兽医学报，2013,33(12):1842-1847. DONG J,YANG X J,MIAO C X,et al.Development and preliminary application of colony multiplex PCR for serotyping of Clostridium perfringens strains[J].Chinese Journal of Veterinary Medicine,2013,33(12):1842-1847.(in Chinese)
[19] ROOD J I,ADAMS V,LACEY J,et al.Expansion of the Clostridium perfringens toxin-based typing scheme[J].Anaerobe,2018,53:5-10.
[20] 郑晓星.产气荚膜梭菌p1c和NetB基因的表达及单克隆抗体制备与鉴定[D].哈尔滨：东北农业大学，2012. ZHENG X X.Expression p1c and NetB of Clostridium perfringens and characterization of monoclonal antibodies against p1c and NetB of proteins[J].Harbin:Northeast Agricultural University,2012.(in Chinese)
[21] 候照峰,冯茜茜,李贝贝,等.鹅源产气荚膜梭菌的分离鉴定及药敏试验[J].中国家禽，2022,44(5):32-37. HOU Z F,FENG X X,LI B B,et al.Isolation,identification and drug sensitivity test of Clostridium perfringens from goose[J].China Poultry,2022,44(5):32-37.(in Chinese)
[22] NCCLS.Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals[J].Approved Standard, 2002，5:M31-A2.
[23] 张娜.羊源产气荚膜梭菌的分离鉴定及耐药性分析[D].杨凌：西北农林科技大学，2019. ZHANG N.Isolation identification and drug resistance analysis of Clostridium perfringens from sheep[D].Yangling：Northwest A&F University,2019.(in Chinese)
[24] 吴克,赵学亮,冯航,等.甘肃省牛和羊源产气荚膜梭菌耐药性分析[J].动物医学进展,2022,43(1):141-144. WU K,ZHAO X L,FENG H,et al.Antimicrobial susceptibility test of Clostridium perfringens from sheep and cattle in Gansu province[J].Progress in Veterinary Medicine,2022,43(1):141-144.(in Chinese)
[25] 罗润波,吴丹,黄家旗,等.青藏高原地区牦牛源产气荚膜梭菌流行病学调查及耐药性分析[J].中国兽医学报,2023,43(9):1851-1858. LUO R B,WU D,HUANG J Q,et al.Epidemiological investigation and drug resistance analysis of Clostridium perfringens yaks on Qinghai-Tibet plateau[J].Chinese Journal of Veterinary Medicine,2023,43(9):1851-1858.(in Chinese)
[26] 黄广明,宝立群,牟和鑫,等.我国地区牧场产气荚膜梭菌感染状况的调查分析[J].中国乳业,2019,12:47-49. HUANG G M,BAO L Q,MOU H X,et al.Investigation and analysis of Clostridium perfringens infection status in China’s regional pastures[J].China Dairy,2019,12:47-49.(in Chinese)
[27] KHAN Z U M.中国和巴基斯坦鸡源产气荚膜梭菌菌株基因型分析及分子流行病学研究[D].北京：中国农业科学院,2020. KHAN Z U M.Profiling & molecular typeing of Clostridium perfringens isolated from broiler chicken in China and Pakistan[D].Beijing：Chinese Academy of Agricultural Sciences,2020.(in Chinese)
[28] 蔡重振,罗润波,黄家旗,等.西藏林芝牦牛源产气荚膜梭菌的分离鉴定及耐药性分析[J].中国兽医学报,2024,44(1):97-106. CAI C Z,LUO R B,HUANG J Q,et al.Isolation,identification and drug resistance analysis of Clostridium perfringens from yaks in Nyingchi,Xizang[J].Chinese Journal of Veterinary Medicine,2024,44(1):97-106.(in Chinese)
[29] 刘红燕,郭亚男,王建东,等.宁夏某牛场致泌乳奶牛急性死亡的产气荚膜梭菌分离鉴定及其药敏试验[J].家畜生态学报,2024,45(6):65-70. LIU H Y,GUO Y N,WANG J D,et al.Isolation and antibiotic susceptibility test of Clostridium perfringens causing acute death of dairy cows in a dairy farm of Ningxia Hui Autonomous Region[J].Journal of Domestic Animal Ecology,2024,45(6):65-70.(in Chinese)
[30] 马紫薇,马学军,马亚楠,等.新疆喀什某规模化安格斯牛场犊牛产气荚膜梭菌的分离鉴定及耐药性分析[J].中国畜牧兽医,2024,51(5):2188-2197. MA Z W,MA X J,MA Y N,et al.Isolation，identification and drug resistance analysis of Clostridium perfringens in calves in a large-scale Angus beef cattle farm in Kashgar,Xinjiang[J].China Animal Husbandry & Veterinary Medicine,2024,51(5):2188-2197.(in Chinese)
[31] KOTTLAHOUSE A,KUMARAGURUBARAN K,VIJAYABALAN D,et al.Toxinotyping and molecular characterization of antimicrobial resistance in Clostridium perfringens isolated from different sources of livestock and poultry[J].Anaerobe,2020,67:102298.
[32] 周增,王迪,高萌萌,等.黑龙江地区“牛猝死症”病原分离鉴定及生物学特性分析[J].中国奶牛,2024,11:28-32. ZHOU Z,WANG D,GAO M M,et al.Isolation，identification and biological characteristics analysis of the pathogen of sudden cattle death syndrome in Heilongjiang region[J].China Dairy Cattle,2024,11:28-32.(in Chinese)
[33] 王冬经,吴金措姆,曾江勇.牦牛源A型产气荚膜梭菌的分离鉴定及生物学特性研究[J].中国畜牧兽医,2024,51(1):229-241. WANG D J,WU J C M,ZENG J Y.Isolation，identification and biological characteristics analysis of Clostridium perfringens type A from yaks[J].China Animal Husbandry & Veterinary Medicine,2024,51(1):229-241.(in Chinese)
[34] 张仕泓,王少林.动物源产气荚膜梭菌耐药性研究进展[J].畜牧兽医学报,2021,52(10):2762-2771. ZHANG S H,WANG S L.Research progress on antimicrobial resistance of Clostridium perfringens of animal origins[J].Acta Veterinaria et Zootechnica Sinica,2021,52(10):2762-2771.(in Chinese)

Investigation of the Pathogen of Calf Diarrhea in Gansu Region and Isolation, Identification and Drug Resistance Analysis of Clostridium perfringens

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