Glanders or farcy, caused by Burkholderia mallei, is an infectious and zoonotic disease of solipeds. Horses, donkeys and mules are the only known natural reservoir of B. mallei. Although glanders has been eradicated from most countries, it has regained the status of a re‐emerging disease because of the numerous recent outbreaks. Pre‐symptomatic or carrier animals are the potential source of infection for the healthy equine population and play a crucial role in the spreading of the infectious agent. Glanders is characterized by ulcerating nodular lesions of the skin and mucous membrane. Generalized symptoms include fever, malaise, depression, cough, anorexia and weight loss. Burkholderia mallei can invade its host through mucous membranes, gastrointestinal tract and the integument. Its virulence mechanisms and pathogenesis are not yet completely understood. A major problem when using serological tests for diagnosing glanders is the occurrence of false‐positive and false‐negative results leading to difficulties in international trade with equids and to the spread of glanders to disease‐free regions. Moreover, poor tests critically result in poor control of disease. These tests are not only incapable of discriminating between B. mallei and B. pseudomallei antibodies, they are also unable to differentiate between malleinized and naturally infected animals. Combined use of both serological and molecular detection methods increases the detection rate of glanders. Countermeasures against glanders include early detection of disease in susceptible animals, stringent quarantine measures, testing and safe destruction of infected carcasses, adequate compensation to the animal owners, disinfection of infected premises and awareness about glanders and the zoonotic implications through veterinary extension services. An account of the clinical picture and successful experimental therapy of spontaneous equine glanders is also given.

The present study was conducted to determine the seroprevalence and identify risk factors associated with brucellosis in humans at high risk in the Potohar plateau of northeastern Pakistan. A total of 262 serum samples were collected from persons of different occupational groups: veterinary personnel, milkers, abattoir workers, livestock farmers, and others (drivers, security guards, housewives). Data related to gender, age, occupation, contact with animals, brucellosis-related symptoms, consumption of raw milk, and geographical region were collected. The Rose Bengal plate test and the serum agglutination test were performed to determine the seroprevalence of brucellosis. The overall seroprevalence was found to be 6.9% (95% confidence interval [CI]: 4.1, 10.6). Real-time polymerase chain reaction assay showed that all cases were affected by Brucella abortus. Individuals who consumed raw milk had higher odds of brucellosis seropositivity. This is the first report of human brucellosis related to B. abortus in high-risk professionals from Pakistan by the combined use of serological and molecular methods.

Brucellosis in livestock causes enormous losses for economies of developing countries and poses a severe health risk to consumers of dairy products. Little information is known especially on camel brucellosis and its impact on human health. For surveillance and control of the disease, sensitive and reliable detection methods are needed. Although serological tests are the mainstay of diagnosis in camel brucellosis, these tests have been directly transposed from cattle without adequate validation. To date, little information on application of real-time PCR for detection of Brucella in camel serum is available. Therefore, this study was performed to compare the diagnostic efficiency of different serological tests and real-time PCR in order to identify the most sensitive, rapid and simple combination of tests for detecting Brucella infection in camels.

The seroprevalence and risk factors of bovine brucellosis were studied at animal and herd level using a combination of culture, serological and molecular methods. The study was conducted in 253 randomly selected cattle herds of the Potohar plateau, Pakistan from which a total of 2709 serum (1462 cattle and 1247 buffaloes) and 2330 milk (1168 cattle and 1162 buffaloes) samples were collected. Data on risk factors associated with seroprevalence of brucellosis were collected through interviews using questionnaires. Univariable and multivariable random effects logistic regression models were used for identifying important risk factors at animal and herd levels. One hundred and seventy (6.3%) samples and 47 (18.6%) herds were seropositive for brucellosis by Rose Bengal Plate test. Variations in seroprevalence were observed across the different sampling sites. At animal level, sex, species and stock replacement were found to be potential risk factors for brucellosis. At herd level, herd size (≥9 animals) and insemination method used were important risk factors. The presence of Brucella DNA was confirmed with a real-time polymerase chain reaction assay (qRT-PCR) in 52.4% out of 170 serological positive samples. In total, 156 (6.7%) milk samples were positive by milk ring test. B. abortus biovar 1 was cultured from 5 positive milk samples. This study shows that the seroprevalence of bovine brucellosis is high in some regions in Pakistan. Prevalence was associated with herd size, abortion history, insemination methods used, age, sex and stock replacement methods. The infected animal may act as source of infection for other animals and for humans. The development of control strategies for bovine brucellosis through implementation of continuous surveillance and education programs in Pakistan is warranted.

Purpose: Pet animals have been considered a potential carrier of clinically important multidrug-resistant Escherichia coli. However, little is known about the role of pets as reservoirs of extended-spectrum β-lactamase (ESBL) producing E. coli in Pakistan. This study was designed to determine the prevalence and genetic relatedness of ESBL-producing multidrug-resistant E. coli in pets, their owners, and veterinary professionals. Methods: A total of 105 fecal samples were collected from dogs, cats, their owners, and veterinary professionals from veterinary clinics. Isolates of ESBL-producing E. coli were subjected to antimicrobial susceptibility testing. The presence of bla _CTX-M genes and CTX-M groups I and II in multidrug-resistant E. coli was detected using PCR. Clonal diversity was checked using BOX-PCR. Results: Of the 105 fecal samples screened, 73 (69.5%) were found to contain ESBL-producing E. coli. The percentage of ESBL-producing E. coli isolates in dogs and dog owners was found to be 81.8% (18/22) and 59% (13/22), respectively. In cats, this percentage was 73.9% (17/23) and in cat owners, 56.5% (13/23). Furthermore, 80% (12/15) of E. coli isolates in veterinary professionals were ESBL producers. Of these 73 ESBL-producing E. coli isolates, 23 isolates exhibited a multidrug-resistant phenotype. The most prevalent multidrug-resistant pattern (17.4%) identified was resistant to ampicillin, cefotaxime, ciprofloxacin, and nitrofurantoin. In the multidrug-resistant E. coli, bla _CTX-M was identified as the most common ESBL-producing genotype (19/23), with bla _CTX-M-1 dominating in all 19 isolates. Furthermore, BOX-PCR analysis exhibited genetically diverse clonal groups among isolates of the CTX-M-1 group. Conclusion: Our results provide important baseline information on the potential burden of multidrug-resistant E. coli among companion animals in Pakistan. Further studies are needed to understand the drivers of antimicrobial resistance at human–animal–environmental intersections.