Brucellosis is a neglected zoonosis that affects animals and people in much of the underdeveloped world. The disease is endemic in cattle in Punjab, India and controlling it is a public health challenge. Dairy farmers and farm labour commonly handle cattle faeces with bare hands and personal protective equipments are not used. No studies have been conducted about the shedding of Brucella species in faeces of sero positive cattle in the state. This study aimed to isolate and identify the Brucella species from faeces of sero positive cattle in Punjab, India. Faecal samples were collected from 350 Brucella sero positive cattle in Ludhiana district of Punjab, India. Isolation was performed using a pre-enriched Brucella selective broth medium as well as Brucella selective medium agar plates containing horse serum and Brucella selective supplements. Isolates were identified using Gram staining technique and rapid slide agglutination test, and then confirmed by using bcsp31 and 16s rRNA genus specific PCR. Isolates were further identified up to species level by using Bruce-Ladder multiplex PCR. Fourteen Brucella species were isolated, all of which showed coccobacilli on gram staining, positive rapid slide agglutination test and amplification of bcsp31 and 16s rRNA genes. Of the 14 isolates, 11 were identified as Brucella abortus and 3 were identified as Brucella melitensis. The study demonstrates that animal faeces could pose a potential risk for animal and human health and faeces of seropositive cattle must be handled with care.

布鲁氏菌病是一种被忽视的人畜共患病，影响许多不发达国家的动物和人类。这种疾病在印度旁遮普邦的牛中流行，控制它是一项公共卫生挑战。奶农和农场工人通常徒手处理牛粪，不使用个人防护装备。尚未对该州血清阳性牛的粪便中布鲁氏菌的脱落情况进行研究。本研究旨在从印度旁遮普邦血清阳性牛的粪便中分离和鉴定布鲁氏菌属。粪便样本采集自印度旁遮普邦卢迪亚纳区的350 头布鲁氏菌血清阳性牛。使用预富集的布鲁氏菌选择性肉汤培养基以及含有马血清和布鲁氏菌选择性补充剂的布鲁氏菌选择性培养基琼脂平板进行分离。使用革兰氏染色技术和快速玻片凝集试验对分离株进行鉴定，然后使用bcsp 31 和 16s rRNA 属特异性 PCR 进行确认。通过使用 Bruce-Ladder 多重 PCR 进一步鉴定分离株至物种水平。分离出14 种布鲁氏菌，革兰氏染色均显示球杆菌，快速玻片凝集试验呈阳性，且bcsp 31 和 16s rRNA 基因扩增。14个分离株中，11个被鉴定为流产布鲁氏菌，3个被鉴定为羊种布鲁氏菌。研究表明，动物粪便可能对动物和人类健康构成潜在风险，血清阳性牛的粪便必须小心处理。