Perceptions and preparedness of veterinarians to combat brucellosis through Brucellosis Control Programme in India.

BACKGROUND AND AIM: Brucellosis caused by bacteria belongs to the genus Brucella is an important zoonosis and constitutes a serious public health hazard worldwide including India. The present study aimed to estimate the knowledge of veterinarians on brucellosis, its public health threat, diagnosis, and vaccination. MATERIALS AND METHODS: This cross-sectional study was conducted during 2013-2015 and 453 veterinarians representing 11 states/Union Territories (UT) of India (Assam, Tripura, Meghalaya, Goa, Karnataka, Madhya Pradesh, Uttar Pradesh, Delhi, Jammu and Kashmir, Tamil Nadu, and Punjab) were interviewed using self-administered questionnaire. RESULTS: Out of 453 veterinarians, 71.74% stated handling of the animals on day-to-day basis and 28.25% were engaged in administration activities. The veterinarians ranked foot-and-mouth disease and brucellosis at the first and fourth ranks among the list of ten economic impacted diseases in the country. A significant association was observed between laboratory confirmation with those who handled brucellosis-suspected cases (p=0.000). Similarly, significant association was noted for the availability of vials/slides (p=0.114), vacutainers (p=0.008), icebox (p=0.103), and refrigerator (p=0.106) for those who preferred laboratory diagnosis. Only 20% of the veterinarians recommended vaccination against bovine brucellosis, and 17% obtained laboratory confirmation for the brucellosis-suspected cases. CONCLUSION: The study highlighted the need for awareness programs, laboratory facilities, veterinary doctors, and protective measures for the veterinarians for combating brucellosis through the control program in the country.

Brucellosis in India: results of a collaborative workshop to define One Health priorities.

Brucellosis is an important zoonosis worldwide. In livestock, it frequently causes chronic disease with reproductive failures that contribute to production losses, and in humans, it causes an often-chronic febrile illness that is frequently underdiagnosed in many low- and middle-income countries, including India. India has one of the largest ruminant populations in the world, and brucellosis is endemic in the country in both humans and animals. In November 2017, the International Livestock Research Institute invited experts from government, national research institutes, universities, and different international organizations to a one-day meeting to set priorities towards a "One Health" control strategy for brucellosis in India. Using a risk prioritization exercise followed by discussions, the meeting agreed on the following priorities: collaboration (transboundary and transdisciplinary); collection of more epidemiological evidence in humans, cattle, and in small ruminants (which have been neglected in past research); Economic impact studies, including cost effectiveness of control programmes; livestock vaccination, including national facilities for securing vaccines for the cattle population; management of infected animals (with the ban on bovine slaughter, alternatives such as sanctuaries must be explored); laboratory capacities and diagnostics (quality must be assured and better rapid tests developed); and increased awareness, making farmers, health workers, and the general public more aware of risks of brucellosis and zoonoses in general. Overall, the meeting participants agreed that brucellosis control will be challenging in India, but with collaboration to address the priority areas listed here, it could be possible.

Seroprevalence of brucellosis in small ruminants in organized and unorganized sectors of Gujarat state, India.

AIM: The present study aimed to study the seroprevalence of brucellosis in small ruminants of Gujarat state, India, using Rose Bengal Plate test (RBPT) and indirect enzyme-linked immunosorbent assay (iELISA). MATERIALS AND METHODS: A total of 2444 sera samples (675 sheep and 1769 goat) from unorganized sector and 1310 sera samples (861 sheep and 449 goat) from seven organized farms were collected for brucellosis screening. RESULTS: In unorganized sector, 23.70% sheep (160/675) and 15.99% goat (283/1769) were positive by RBPT and 24.44% sheep (165/675) and 17.24% goat (305/1769) by iELISA. The organized sector samples showed higher seroprevalence in goat (7.79 %, 35/449) than sheep (4.06 %, 35/861) by RBPT. Similarly, in iELISA, goat samples showed a higher seroprevalence (9.35%, 42/449) compared to sheep (7.50%, 65/861). The diagnostic sensitivity and specificity of RBPT with ELISA were 88.69% and 99.65%, respectively, and showed a significant difference (p≤0.0001). The Chi-square analysis revealed a significant difference in seroprevalence between sectors (p≤0.01) and species (p≤0.01). CONCLUSION: The seroprevalence of brucellosis in small ruminants of Gujarat was investigated and showed a higher prevalence of brucellosis and warrants the implementation of proper preventive measures.

Evaluation of lateral flow assay as a field test for investigation of brucellosis outbreak in an organized buffalo farm: A pilot study.

AIM: The aim was to evaluate lateral flow assay (LFA) as a field test for investigation of brucellosis outbreak in organized buffalo farm. MATERIALS AND METHODS: A total of 153 serum samples were tested to detect the presence of brucella antibodies by LFA and three other serological tests i.e. rose bengal plate test (RBPT), protein G based indirect enzyme-linked immunoassay (iELISA), and competitive ELISA (cELISA). The performances of LFA and other serological tests were evaluated using OIE complaint cELISA as the gold standard. RESULTS: Serological tests revealed 50% of the animals were seropositive for Brucella antibodies and correlated with clinical history of abortions, infertility, and productive failures. The newly developed assay showed 87.1% and 92.6% sensitivity and specificity, which was even higher than the specificity of RBPT. CONCLUSIONS: The investigation proved the potential usefulness of LFA for field diagnosis of brucellosis in the regions where laboratory facilities are limited.

Staphylococcus aureus spa type t267, clonal ancestor of bovine subclinical mastitis in India.

AIM: To evaluate the virulence determinants and genetic diversity of Staphylococcus aureus from bovine subclinical mastitis milk. METHODS AND RESULTS: PCR detection of virulence genes was performed for 173 Staph. aureus from bovine subclinical mastitis milk. Further, genetic diversity was analysed by agr and spa typing followed by pulsed field gel electrophoresis (PFGE) of selected isolates. Screening of virulence genes (n = 19) showed the adherence genes viz. fnbA, clfA, fnbB and cna in 98·8, 97·1, 68·8 and 28·3 percentage of isolates, respectively, and 80 strains (46·24%) positive for enterotoxin genes were distributed as 23 toxinotypes, of which, 5 genotypes contained a single gene and the rest comprised of multiple toxin genes. Out of agr type-1 (87·3%), 74·2 per cent belonged to the three predominant spa types. Of 27 spa types, 11 were identified for the first time. The predominant spa types were t267 (N =44), t359 (N = 42) and t6877 (N =29), which together accounts to 66·5 per cent of isolates. PFGE analysis of isolates (N = 45) covering all the spa types revealed mostly similar or closely related pulsotypes. Local emergence of spa type t6877 in herd-dependant manner was observed. spa sequence-based phylogenetic analysis suggested t267 as the ancestral clone of t359, t6877 and other spa types except two. CONCLUSION: Heterogenous virulence profile of the isolates had no significant association with the genotype. High prevalence of agr group I reaffirms their association with persistent subclinical mastitis. The spa type t267 appears to be the ancestral clone endemic in the region causing subclinical mastitis. In addition, few new spa types have emerged in the geographic region. SIGNIFICANCE AND IMPACT OF STUDY: Gives an insight into the genetic and evolutionary behaviour of Staph. aureus associated with bovine subclinical mastitis in India. The study would pave the way for devising effective control strategy for bovine mastitis in Indian context.

Multiplex PCR assay for species identification of bovine mastitis pathogens.

AIM: To develop and evaluate a multiplex PCR (mPCR) assay for simultaneous detection of 10 bacterial species causing bovine mastitis namely, Staphylococcus aureus, Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus sciuri, Staphylococcus haemolyticus, Staphylococcus simulans, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis and Escherichia coli in milk. METHODS AND RESULTS: A two-tube mPCR assay was developed. The accuracy of the mPCR was evaluated using 56 standard reference strains and 705 strains comprising of E. coli (n = 99), staphylococci (n = 522) and streptococci (n = 84). The threshold of detection of the mPCR assay was 10 fg of genomic DNA and <10(3) CFU ml(-1). A comparative evaluation of mPCR with culture method using 115 milk samples from subclinical mastitis showed mPCR to be more efficacious. Subsequently, the mPCR showed successful detection of target bacteria, when applied directly for the assessment of 36 bulk milk samples. CONCLUSION: The developed mPCR assay was found to be simple, rapid, reliable and specific in species identification of 10 bacteria at a time. SIGNIFICANCE AND IMPACT OF THE STUDY: The assay will be useful for the detection of mastitis, testing bacteriological safety of milk and for species level differentiation. The assay will be of value in the dairy sector for diagnosis and research. The early and accurate identification of pathogens will enable timely interventions for the treatment and control of bovine mastitis.

Investigation of haemorrhagic enteritis in pygmy hogs (Sus salvanius) from India.

The pygmy hog is a representative of the smallest and rarest wild species of known living Suidae. This paper reports the investigation of haemorrhagic enteritis encountered amongst the pygmy hogs at the Research and Breeding Centre of the Pygmy Hog Conservation Programme, Guwahati, Assam, India. Three out of 68 pygmy hogs died of enteric infection. Post-mortem examination and bacteriological investigation of two out of the three animals that died revealed clostridial infection. The isolates harboured two plasmids of molecular weight 42.8 kilobases (kb) and 51.9 kb. Clostridium perfringens Type A positive for the beta2 toxin (cpb2) gene was detected by polymerase chain reaction. Sequence analysis of the partial alpha toxin (cpa) gene showed 98% to 100% homology with isolates from different geographical locations.