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肯尼亚奈若比县穆库雷尼次县从奶山羊中分离的乳腺炎病原菌的流行情况和耐药谱。

Prevalence and antimicrobial resistance profiles of mastitis causing bacteria isolated from dairy goats in Mukurweini Sub-County, Nyeri County, Kenya.

机构信息

Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya.

Department of Medical Microbiology, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya.

出版信息

Vet Med Sci. 2024 May;10(3):e1420. doi: 10.1002/vms3.1420.

Abstract

BACKGROUND

Ruminant mastitis continues to be a cause of economic losses in the dairy industry and remains a major public health hazard globally.

OBJECTIVES

This cross-sectional study was carried out in Mukurweini Sub-County of Nyeri County, Kenya, to investigate the prevalence of bacteria causing mastitis, risk factors associated with goat mastitis and the antibiotic resistance profiles of bacteria isolated from the goat milk.

METHODS

Farm level data on risk factors for mastitis was obtained from 56 farmers using a semi structured questionnaire. A total of 189 goat milk samples were collected. The goat's udder was observed for signs of clinical mastitis and the California Mastitis Test (CMT) used to test the milk for sub-clinical mastitis. All samples were then cultured for morphological identification of bacteria and strain typing by Matrix Assisted Laser Desorption/Ionization (MALDI)-Time of Flight (ToF) technique. Antimicrobial susceptibility of the isolated Staphylococcus aureus, coagulase-negative Staphylococcus (CoNS), Escherichia coli, Klebsiella oxytoca, Pseudomonas spp., Enterobacter spp., Proteus vulgaris and Escherichia vulneris to eight commonly used antibiotics was done by the disc diffusion method and validated by determining the presence of antibiotic resistance genes (mecA and blaTEM) using polymerase chain reaction method.

RESULTS

The prevalence of clinical mastitis was 1.1% (2/189) while that of sub-clinical mastitis was 84.7% (160/189). Higher (p < 0.05) prevalence of mastitis was observed in goats whose houses were cleaned fortnightly and in cases where farmers used same towel to dry different does' udders during the milking process. Thirteen different bacterial species were isolated from the milk samples and identified by MALDI-ToF, and these included S. aureus (22.0%), CoNS (20.3%), E. coli (18.1%), Pseudomonas spp. (14.3%), Enterobacter spp. (10.4%), K. oxytoca (6.0%), E. vulneris (1.7%), P. vulgaris (1.7%), Raoutella ornithinolytica (1.7%), Stenotrophomonas maltophilia (1.1%), Pantoea agglomerans (1.1%), Serratia marcescens (1.1%) and Cedeceas spp. (0.6%). One hundred pathogenic bacterial isolates were randomly selected and tested for antibiotic sensitivity to eight antibiotics out of which S. aureus were 97.5% resistant to Oxacillin and 100% sensitive to Ciprofloxacin. The CoNSs were 100% resistant to Oxacillin and 100% sensitive to Ciprofloxacin. E. coli were 93.9% resistant to Oxacillin, 69.7% sensitive to Ciprofloxacin and 87.9% sensitive to both Amoxicillin/Clavulanic acid and Meropenem. The antimicrobial resistant genes detected in S. aureus and E. coli were mecA [66.7%, 0%], and bla [20% and 78.3%], respectively.

CONCLUSION

In conclusion, the study showed that most of the does were affected by subclinical mastitis with the main causative bacteria being Staphylococci spp. and coliforms. Farmers need to be trained on improved control of mastitis by adoption of good milking practices and use of CMT kit for early detection of mastitis. Occurrence of multidrug resistance by key mastitis causing pathogens was shown to be prevalent and therefore there is need for development of intervention strategies.

摘要

背景

反刍动物乳腺炎仍然是奶牛养殖业的经济损失原因,并仍然是全球主要的公共卫生危害。

目的

本横断面研究在肯尼亚恩耶里县穆库雷维尼次县进行,旨在调查引起乳腺炎的细菌流行情况、与山羊乳腺炎相关的风险因素以及从山羊奶中分离的细菌的抗生素耐药谱。

方法

使用半结构化问卷从 56 位农民那里获得有关乳腺炎风险因素的农场水平数据。共采集了 189 份山羊奶样本。观察山羊乳房是否有临床乳腺炎的迹象,并使用加利福尼亚乳腺炎测试(CMT)测试牛奶是否有亚临床乳腺炎。然后,所有样本均进行形态学鉴定,并通过基质辅助激光解吸/电离(MALDI)-飞行时间(ToF)技术进行菌株分型。采用纸片扩散法测定分离的金黄色葡萄球菌、凝固酶阴性葡萄球菌(CoNS)、大肠杆菌、产酸克雷伯菌、假单胞菌、肠杆菌、普通变形杆菌和脆弱拟杆菌对 8 种常用抗生素的药敏性,并通过聚合酶链反应法检测抗生素耐药基因(mecA 和 blaTEM)的存在来验证。

结果

临床乳腺炎的患病率为 1.1%(2/189),而亚临床乳腺炎的患病率为 84.7%(160/189)。在房屋每两周清洁一次且农民在挤奶过程中使用同一毛巾擦干不同母羊乳房的情况下,乳腺炎的患病率更高(p<0.05)。从牛奶样本中分离出了 13 种不同的细菌,通过 MALDI-ToF 鉴定,包括金黄色葡萄球菌(22.0%)、凝固酶阴性葡萄球菌(20.3%)、大肠杆菌(18.1%)、假单胞菌(14.3%)、肠杆菌(10.4%)、产酸克雷伯菌(6.0%)、脆弱拟杆菌(1.7%)、普通变形杆菌(1.7%)、鸟氨酸罗特氏菌(1.7%)、嗜麦芽窄食单胞菌(1.1%)、成团泛菌(1.1%)、黏质沙雷氏菌(1.1%)和食酸菌属(0.6%)。随机选择了 100 个致病细菌分离株,测试了 8 种抗生素的药敏性,其中金黄色葡萄球菌对苯唑西林的耐药率为 97.5%,对环丙沙星的敏感率为 100%。CoNS 对苯唑西林的耐药率为 100%,对环丙沙星的敏感率为 100%。大肠杆菌对苯唑西林的耐药率为 93.9%,对环丙沙星的敏感率为 69.7%,对阿莫西林/克拉维酸和美罗培南的敏感率为 87.9%。金黄色葡萄球菌和大肠杆菌中检测到的抗生素耐药基因分别为 mecA[66.7%,0%]和 bla[20%和 78.3%]。

结论

总之,该研究表明,大多数母羊都受到亚临床乳腺炎的影响,主要的病原体是葡萄球菌和大肠埃希菌。农民需要通过采用良好的挤奶方法和使用 CMT 试剂盒进行早期乳腺炎检测,接受更好的乳腺炎控制培训。主要乳腺炎病原体表现出多药耐药性的发生非常普遍,因此需要制定干预策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/95fc/10976424/8bf55d081a3a/VMS3-10-e1420-g003.jpg

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