Mapping Heterogeneous Population Structure of Mannheimia haemolytica Associated with Pneumonic Infection of Sheep in Southern State Karnataka, India.

Mannheimia haemolytica is recognized as principal pathogen associated with pneumonic pasteurellosis leading to huge economic losses to small ruminant farmers. Even though the disease causes huge economic losses, epidemiology of M. haemolytica is less studied, hindering the formulation of effective control strategies. Current study aimed to highlight molecular characterisation of M. haemolytica strains isolated from ovine pneumonic infection. M. haemolytica 27 isolates with two reference strains were characterised using capsular and virulence gene typing, multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) methods. M. haemolytica serotype A2 recognized as predominant serotype (74%) followed by A6 (11%) and A1 (5%) serotypes. Virulence gene profiling by PCRs showed dominance of all five virulent genes [such as adh and gcp (100% each)] followed by gs60 (88.8%), lktC (85.2%), tbpB (51.9%) and least nmaA gene (14.8%). MLST profiling delineated M. haemolytic isolates into 11 sequence types (STs) with most prevalent being ST37 (27.9%) and ST16 (23%) and nine new STs (ST37, 38, 39, 40, 41, 42, 47, 48, and 49). These new STs did not belong to any of the three clonal complexes (CC4, CC8 and CC28). ST16 was exclusively noted in A1 and A6 serotypes. Amongst 25 isolates, 22 pulsotypes (GD 0.88) recorded indicated variability of the M. haemolytica isolates in PFGE analysis. In conclusion, the study suggested dominance of M. haemolytica serotype A2 harbouring different virulent genes, diverse STs and pulsotypes responsible for pneumonic pasteurellosis frequently encountered in sheep.

Genetic insights of antibiotic resistance, pathogenicity (virulence) and phylogenetic relationship of Escherichia coli strains isolated from livestock, poultry and their handlers - a one health snapshot.

BACKGROUND: Pathogenic and non-pathogenic strains of Escherichia coli harbouring antibiotic resistance genes (ARGs) from any source (clinical samples, animal settings, or environment) might be transmitted and contribute to the spread and increase of antibiotic resistance in the biosphere. The goal of this study was to investigate the genome to decipher the repertoire of ARGs, virulence genes carried by E. coli strains isolated from livestock, poultry, and their handlers (humans), and then unveil the genetic relatedness between the strains. METHODS: Whole genome sequencing was done to investigate the genetic makeup of E. coli isolates (n = 20) [swine (n = 2), cattle (n = 2), sheep (n = 4), poultry (n = 7), and animal handlers (n = 5)] from southern India. The detection of resistome, virulome, biofilm forming genes, mobile genetic elements (MGE), followed by multilocus sequence typing (MLST) and phylogenetic analyses, were performed. RESULTS: E. coli strains were found to be multi drug resistant, with a resistome encompassing > 20 ARGs, the virulome-17-22 genes, and > 20 key biofilm genes. MGE analysis showed four E. coli isolates (host: poultry, swine and cattle) harbouring composite transposons with ARGs/virulence genes (blaTEM, dfr, qnr/nleB, tir, eae,and esp) with the potential for horizontal transfer. MLST analyses revealed the presence of ST937 and ST3107 in both livestock/poultry and their handlers. Phylogenomic analyses with global E. coli isolates (human/livestock/poultry hosts) showed close relatedness with strains originating from different parts of the world (the United States, China, etc.). CONCLUSION: The current study emphasizes the circulation of strains of pathogenic sequence types of clinical importance, carrying a diverse repertoire of genes associated with antibiotic resistance, biofilm formation and virulence properties in animal settings, necessitating immediate mitigation measures to reduce the risk of spread across the biosphere.

Prevalence and epidemiological characteristics of methicillin-resistant Staphylococcus aureus isolated from cattle in Bangalore India as a part of the One Health approach.

In India, limited studies are available on the epidemiological aspects of methicillin-resistant Staphylococcus aureus (MRSA) infections in both animal and human settings. Herein, we investigated the prevalence, antimicrobial resistance profile and molecular characteristics of MRSA isolates recovered from cattle using the One Health approach. Out of 66 mecA-positive staphylococci, species-specific multiplex PCR detected 24 % (n=16) of isolates as MRSA. Maximum antibiotic resistance was seen against cloxacillin (94 %, n=15) and least for enrofloxacin and cephalothin (each 13 %, n=2). Overall, 13 % (n=2) of MRSA isolates were multidrug-resistant. Molecular characterization by SCCmec typing identified 88 % (n=14) of MRSA isolates as type V. Twelve isolates (75 %) belonged to novel spa-type t17242, of which 67 % (n=8) belonged to agr type I. MLST analysis revealed ST 1687 (50 %, n=8) as the most predominant sequence type. Circulation of different MRSA clones among the cattle populace offers a risk of transmission to humans through direct contact, food chain or environmental contamination. Thus, continuous monitoring of MRSA strains is imperative for early diagnosis and for establishing effective treatment strategies to restrain the disease burden caused by MRSA infections.

Analyses of Extended-Spectrum-ß-Lactamase, Metallo-ß-Lactamase, and AmpC-ß-Lactamase Producing Enterobacteriaceae from the Dairy Value Chain in India.

The consumption of milk contaminated with antibiotic-resistant bacteria poses a significant health threat to humans. This study aimed to investigate the prevalence of Enterobacteriaceae producing ß-lactamases (ESBL, MBL, and AmpC) in cow and buffalo milk samples from two Indian states, Haryana and Assam. A total of 401 milk samples were collected from dairy farmers and vendors in the specified districts. Microbiological assays, antibiotic susceptibility testing, and PCR-based genotyping were employed to analyze 421 Gram-negative bacterial isolates. The overall prevalence of ß-lactamase genes was 10% (confidence interval (CI) (7-13)), with higher rates in Haryana (13%, CI (9-19)) compared to Assam (7%, CI (4-11)). The identified ß-lactamase genes in isolates were blaCMY, blaMOX, blaFOX, blaEBC, and blaDHA, associated with AmpC production. Additionally, blaCTX-M1, blaSHV, and blaTEM were detected as ESBL producers, while blaVIM, blaIMP, blaSPM, blaSIM, and blaGIM were identified as MBL producers. Notably, Shigella spp. were the dominant ß-lactamase producers among identified Enterobacteriaceae. This study highlights the presence of various prevalent ß-lactamase genes in milk isolates, indicating the potential risk of antimicrobial-resistant bacteria in dairy products. The presence of ß-lactam resistance raises concern as this could restrict antibiotic options for treatment. The discordance between genotypic and phenotypic methods emphasizes the necessity for comprehensive approaches that integrate both techniques to accurately assess antibiotic resistance. Urgent collaborative action incorporating rational and regulated use of antibiotics across the dairy value chain is required to address the global challenge of ß-lactam resistance.

Management of bovine brucellosis in organized dairy herds through the identification of risk factors: A cross-sectional study from Karnataka, India.

Background and Aim: Brucellosis is an infectious disease caused by Brucella species. This study aimed to identify the risk factors associated with bovine brucellosis seropositivity in organized dairy farms to control the disease in unvaccinated adult bovine herds in Karnataka, India. Materials and Methods: In total, 3610 samples (3221 cattle and 389 buffaloes) were subjected to parallel testing using the Rose Bengal plate test and protein G-based enzyme-linked immunosorbent assay, followed by analyses of animal- and farm-level epidemiological datasets to identify the risk factors. Results: The apparent brucellosis prevalence at the animal level was higher in buffaloes (8.2%, 95% confidence interval [CI] = 5.9-11.4) than in cattle (6.1%, 95% CI = 5.3-7.0). In a multivariable logistic model, animals calved 3-5 times (odds ratio [OR] = 2.22, 95% CI = 1.50-3.1, reference [ref]: animals calved <2 times); animals with a history of abortion (OR = 54.73, 95% CI = 33.66-89.02), repeat breeding (OR = 19.46, 95% CI = 11.72-32.25), and placental retention (OR = 13.94, 95% CI = 4.92-39.42, ref: no clinical signs); and dogs on farms (OR = 2.55, 95% CI = 1.48-4.40, ref: absence of dogs); disposal of aborted fetus in open fields (OR = 4.97, 95% CI = 1.93-12.84) and water bodies (OR = 2.22, 95% CI = 1.50-3.1, ref: buried); purchase of animals from other farms (OR = 6.46, 95% CI = 1.01-41.67, ref: government farms); hand milking (OR = 1.98, 95% CI = 1.02-10.0, ref: machine milking); and use of monthly veterinary services (OR = 3.45, 95% CI = 1.28-9.29, ref: weekly services) were considered significant risk factors for brucellosis in organized bovine herds (p < 0.01). Conclusion: The study identified that the animals calved 3-5 times or with a history of abortion/repeat breeding/placental retention, and disposal of aborted fetus in open fields/water bodies as the potential risk factors for bovine brucellosis. These risk factors should be controlled through the implementation of best practices to reduce the brucellosis burden in bovine farms.

Characterization of humoral and cellular immune responses elicited by reduced doses of Brucella abortus S19 (calfhood) vaccine in cattle calves of India.

Brucella abortus S19 vaccine is a stable attenuated smooth strain, globally used as calfhood vaccine for the prevention of bovine brucellosis. Various agencies demonstrated different doses for vaccinating cattle and buffalo calves leading to ambiguity in selecting a suitable immune vaccine dose. The current study aimed at evaluating four graded doses of S19 vaccine to arrive at the dose which could produce comparable effectiveness as that of full dose prescribed by Indian Pharmacopeia among the Indian calves. Four vaccine doses of which the first dose consisted of full dose (40 × 109 CFU/dose) and the other three were 1/10th, 1/20th, 1/100th reduced doses along with control were tested. Each vaccine dose was administered to 13 cattle calves of 4-5 months of age maintained in separate groups. The blood samples were collected on 0 to 240 days post-vaccination (DPV) at the intervals of 0, 14, 28, 45, 60, 90, 150, 180 and 240 for assessment of vaccine-induced innate, humoral and cell-mediated immune responses. The sero-conversion of all vaccinated animals on DPV 45 and persistence of antibody till DPV 240 were noticed. No significant differences were observed in antibody response between animal groups that received full and 1/10th reduced doses. Innate and cell-mediated response by IL-6, TNF-α¸ IFN-γ, CD4+ and CD8+ cell counts showed dose-dependent responses with no significant difference between full dose and 1/10th reduced doses. The results suggest a possible one log reduction of full dose without compromising immune responses to aid larger vaccination coverage for creating herd immunity.

Evaluation of an in-house IgM/IgG lateral flow assay for serodiagnosis of human brucellosis.

This study aimed to evaluate the diagnostic efficacy of an in-house lateral flow assay (LFA) for the detection of IgM/IgG anti-Brucella antibodies for rapid serodiagnosis of human brucellosis. Three groups of sera samples including 476 from high-risk individuals, 27 from culture-confirmed patients, and 43 from healthy blood donors were used for evaluation of LFA. In comparison with iELISA, the sensitivity, specificity, and accuracy of LFA were >95%, >99%, and 99% respectively. Considering the very good agreement, accuracy, simplicity, and rapidity, LFAs might be useful as a point of care test for the diagnosis of human brucellosis in resource-limited laboratories.

Molecular Characterization of Methicillin-Resistant Staphylococci from the Dairy Value Chain in Two Indian States.

Bovine milk and milk products may contain pathogens, antimicrobial resistant bacteria, and antibiotic residues that could harm consumers. We analyzed 282 gram-positive isolates from milk samples from dairy farmers and vendors in Haryana and Assam, India, to assess the prevalence of methicillin-resistant staphylococci using microbiological tests, antibiotic susceptibility testing, and genotyping by PCR. The prevalence of genotypic methicillin resistance in isolates from raw milk samples was 5% [95% confidence interval, CI (3-8)], with 7% [CI (3-10)] in Haryana, in contrast to 2% [CI (0.2-6)] in Assam. The prevalence was the same in isolates from milk samples collected from farmers [5% (n = 6), CI (2-11)] and vendors [5% (n = 7), CI (2-10)]. Methicillin resistance was also observed in 15% of the isolates from pasteurized milk [(n = 3), CI (3-38)]. Two staphylococci harboring a novel mecC gene were identified for the first time in Indian dairy products. The only SCCmec type identified was Type V. The staphylococci with the mecA (n = 11) gene in raw milk were commonly resistant to oxacillin [92%, CI (59-100)] and cefoxitin [74%, CI (39-94)], while the isolates with mecC (n = 2) were resistant to oxacillin (100%) only. All the staphylococci with the mecA (n = 3) gene in pasteurized milk were resistant to both oxacillin and cefoxitin. Our results provided evidence that methicillin-resistant staphylococci occur in dairy products in India with potential public health implications. The state with more intensive dairy systems (Haryana) had higher levels of methicillin-resistant bacteria in milk.

Genetic characterization and comparative genomics of a multi drug resistant (MDR) Escherichia coli SCM-21 isolated from a subclinical case of bovine mastitis.

Escherichia coli is one of the major pathogens causing mastitis that adversely affects the dairy industry worldwide. This study employed whole genome sequence (WGS) approach to characterize the repertoire of antibiotic resistance genes (resistome), virulence genes (virulome), phylogenetic relationship and genome wide comparison of a multi drug resistant (MDR) E. coli(SCM-21) isolated from a case of subclinical bovine mastitis in Bangalore, India. The genome of E. coli SCM- 21 was found to be of 4.29 Mb size with 50.6% GC content, comprising a resistome of 22 genes encoding beta-lactamases (blaTEM,blaAmpC), polymyxin resistance (arnA) and various efflux pumps (acr, ade, emr,rob, mac, mar, rob), attributing to the bacteria's overall antibiotic resistance genetic profile. The virulome of E. coli SCM-21 consisted of genes encoding different traits [adhesion (ecp, fim, fde), biofilm formation (csg) and toxin production (ent, esp, fep, gsp)], necessary for manifestation of the infection. Phylogenetic relationship of E. coli SCM- 21 with other global E. coli strains (n = 4867) revealed its close genetic relatedness with E. coli strains originating from different hosts of varied geographical regions [human (Germany) bos taurus (USA, Belgium and Scotland) and chicken (China)]. Further, genome wide comparative analysis with E. coli (n = 6) from human and other animal origins showed synteny across the genomes. Overall findings of this study provided a comprehensive insight of the hidden genetic determinants/power of E. coli SCM-21 that might be responsible for manifestation of mastitis and failure of antibiotic treatment. Aforesaid strain forms a reservoir of antibiotic resistance genes (ARGs) and can integrate to one health micro biosphere.

Countrywide cross-sectional study of swine brucellosis sero-prevalence in Indian subcontinent during 2018-2019.

Brucellosis in swine is a contagious disease with greater zoonotic potential caused by Brucella suis. The study describes PAN India swine brucellosis sero-prevalence in 5431 stratified random serum samples collected during 2018-2019 from 26 out of 29 states and two out of seven union territories. The serum samples were tested for anti-Brucella antibodies by indirect ELISA and overall, 4.33% apparent prevalence (AP) was recorded. The AP is ≥ 10% in five states among 26 states, P ≥ 50% in four districts out of 117 districts screened and cent percent prevalence in two epi units out of 264 sampled. Significantly high seropositivity (p < 0.05) in male (6.08%) than female pigs (3.46%) and in ≥ 24-month-old pigs indicated older and male pigs as potential carriers of the disease. The study recorded endemicity of the swine brucellosis in few regions of India requiring periodical surveillance for control of the disease. Brucella testing of boars before breeding and awareness among farmers and veterinarians will aid in reduction of disease burden in the absence of vaccination policy.

Occurrence and characterization of genetic determinants of ß-lactam-resistance in Escherichia coli clinical isolates.

ß-lactamase mediated resistance in Escherichia coli is a significant problem that requires immediate attention. Herein, we aim to characterize and understand the dynamics of the genetic determinants of ß-lactam resistance (i.e. ESBL, AmpC, and MBL) in E. coli. Out of 203 E. coli isolates, genetic determinants of ß-lactam resistance were identified in 50% (n = 101) of isolates. ESBL, AmpC, and MBL resistance determinants were detected in 78%, 40%, and 18% of isolates, respectively with blaCTX-M group 4 (48%), blaCMY (40%), and blaSIM (33%) as the most prevalent ß-lactam resistance genes. Among these isolates, 45% harbored plasmid replicon types, with L/M (40%) and Y (33%) as the most dominant replicon types. Integrons were detected in 40% of such isolates, with Class-1 and Class-3 representing 62% and 55%, respectively. Overall, we observed high rate of genetic determinants of ß-lactam-resistance in E. coli isolates recovered from patients in clinical settings. The co-occurrence of antimicrobial resistance genes and mobile genetic elements in a high percentage of isolates is a major concern and relates to complex resistance mechanisms. To combat the serious threat of antimicrobial resistance, it is imperative to develop strategies for robust surveillance and understand the molecular basis of resistance acquisition and transmission.

Estimates of the Economic Cost Caused by Five Major Reproductive Problems in Dairy Animals in Assam and Bihar, India.

Reproductive problems in dairy animals reduce fertility, prevent conception, create problems in the delivery of healthy calves, lead to postpartum complications, increase inter-calving periods, reduce milk yield, and lower overall lifetime productivity. This study aimed at understanding the incidence of reproductive problems and the cost caused by these. The study covered 954 dairy animals in Bihar and 1348 dairy animals in Assam that were selected using a multi-stage random sampling method. The costs were calculated as the sum of income losses and expenditures incurred. The major cost incurred resulted from extended calving intervals (46.1% of the total cost), followed by loss through salvage selling (38.1%), expenditure for treatment of repeat breeders (5.9%), loss of milk production (5.3%) and expenditure for extra inseminations (2.0%). About one fifth of the selected reproductive problems were left untreated. The estimated cost of reproductive problems was Indian Rupees (INR) 2424.9 (USD 36.1) per dairy animal per year (of the total dairy animal population) which represented approximately 4.1% of the mean value loss of dairy animals (INR 58,966/USD 877) per year. Reproductive problems were significantly (p < 0.001) higher among improved (exotic breed or cross-bred) dairy animals than indigenous (native breed or nondescript indigenous) dairy animals. The study suggests that with the increase of improved dairy animal population, the loss may further increase. The study concludes that any economic estimation of reproduction problems based on aetiology without confirmatory diagnoses could be highly misleading because of the complex nature of the problems.

Understanding Antibiotic Usage on Small-Scale Dairy Farms in the Indian States of Assam and Haryana Using a Mixed-Methods Approach-Outcomes and Challenges.

The use and misuse of antibiotics in both humans and animals contributes to the global emergence of antimicrobial resistant (AMR) bacteria, a threat to public health and infection control. Currently, India is the world's leading milk producer but antibiotic usage within the dairy sector is poorly regulated. Little data exists reflecting how antibiotics are used on dairy farms, especially on small-scale dairy farms in India. To address this lack of data, a study was carried out on 491 small-scale dairy farms in two Indian states, Assam and Haryana, using a mixed method approach where farmers were interviewed, farms inspected for the presence of antibiotics and milk samples taken to determine antibiotic usage. Usage of antibiotics on farms appeared low only 10% (95% CI 8-13%) of farmers surveyed confirmed using antibiotics in their dairy herds during the last 12 months. Of the farms surveyed, only 8% (6-11%) had milk samples positive for antibiotic residues, namely from the novobiocin, macrolides, and sulphonamide classes of antibiotics. Of the farmers surveyed, only 2% (0.8-3%) had heard of the term "withdrawal period" and 53% (40-65%) failed to describe the term "antibiotic". While this study clearly highlights a lack of understanding of antibiotics among small-scale dairy farmers, a potential factor in the emergence of AMR bacteria, it also shows that antibiotic usage on these farms is low and that the possible role these farmers play in AMR emergence may be overestimated.

Comparative evaluation of the immunodominant proteins of Brucella abortus for the diagnosis of cattle brucellosis.

BACKGROUND AND AIM: The present serodiagnosis of brucellosis in livestock is based on the whole cell or smooth lipopolysaccharide of the Brucella organism in which specificity is hampered by the cross-reactivity, especially with the antibodies against Yersinia enterocolitica O:9 organism. The problem can be addressed by screening for better immunodominant antigens. Hence, the present study was undertaken to screen protein antigens of Brucella abortus for their diagnostic potential in cattle brucellosis. MATERIALS AND METHODS: Protein antigens of B. abortus (n=10) non-reactive to antibodies against Y. enterocolitica O:9 were selected, expressed in Escherichia coli, assessed the reactivity of expressed recombinant proteins by Western blot, standardized indirect-enzyme-linked immunosorbent assay (ELISA) for detecting Brucella antibodies in cattle serum, and comparative evaluation was done. RESULTS: All the selected protein antigens were expressed and in the Western blot with Brucella antibodies positive cattle serum, six recombinant (Brucella protein 26 [BP26], Cu-Zn Superoxide dismutase [SodC], B. abortus I-1885, Serine protease, Bacterioferritin, and Brucella Lumazine Synthase [BLS]) proteins showed reaction whereas none of the proteins showed reactivity with Brucella negative cattle serum. ELISA has been done using known Brucella positive and negative cattle sera samples (n=113 each) in which the performance of recombinant proteins in diagnosing brucellosis was in the order of BP26 > BLS > SodC followed by rest of the proteins. BP26 based ELISA was found to be better with area under the curve as 0.953, and diagnostic sensitivity, diagnostic specificity, and Youden's index of 90.27%, 95.58%, and 0.8584, respectively, with the excellent agreement (k=0.85). CONCLUSION: BP26 could be a potential diagnostic antigen among the immunodominant proteins of B. abortus in ruling out Y. enterocolitica O:9 infection while diagnosing brucellosis in cattle herds.

Seroprevalence and Risk Factors of Brucella Infection in Dairy Animals in Urban and Rural Areas of Bihar and Assam, India.

This study assessed seropositivity of Brucella infection in dairy animals and risk factors associated with it. The cross-sectional study used multi-stage, random sampling in the states of Bihar and Assam in India. In total, 740 dairy animals belonging to 534 households of 52 villages were covered under this study. Serological testing was conducted by indirect enzyme-linked immunosorbent assay (iELISA). Animal-level Brucella seropositivity was found to be 15.9% in Assam and 0.3% in Bihar. Seropositivity in urban areas (18.7%) of Assam was found to be higher than in rural areas (12.4%). Bihar was excluded from the risk factor analysis, as only one Brucella seropositive sample was detected in the state. A total of 30 variables were studied for assessing risk factors, of which 15 were selected for multivariable regression analyses following a systematic process. Finally, only three risk factors were identified as statistically significant. It was found that animals belonging to districts having smaller-sized herds were less likely (p < 0.001) to be Brucella seropositive than animals belonging to districts having larger-sized herds. Furthermore, the chance of being Brucella seropositive increased (p = 0.007) with the increase in age of dairy animals, but decreased (p = 0.072) with the adoption of artificial insemination (AI) for breeding. We speculated that the identified risk factors in Assam likely explained the reason behind lower Brucella seropositivity in Bihar, and therefore any future brucellosis control program should focus on addressing these risk factors.

Animal disease surveillance: Its importance & present status in India.

Animal disease surveillance encompasses systematic collection of long-term data on disease events, risk factors and other relevant parameters followed by analyzing the same with reference to temporal and spatial characteristics to arrive at a conclusion so that necessary preventive measures can be taken. In India, the animal disease surveillance is done through National Animal Disease Reporting System, which is a web-based information technology system for disease reporting from States and Union Territories with the aim to record, monitor livestock disease situation and to initiate the preventive and curative action in a swift manner during disease emergencies. National Animal Disease Referral Expert System is a dynamic geographic information system and remote sensing-enabled expert system that captures an incidence of 13 economically important livestock diseases from all over the country and also provides livestock disease forecasting. The laboratories under State and Central governments, several research institutes under the Indian Council of Agricultural Research and veterinary colleges are involved in livestock disease diagnosis including zoonotic diseases. An integrated surveillance system is necessary for early detection of emerging/zoonotic diseases in humans. This review provides information on disease reporting and surveillance systems in animal health sector and the need for One Health approach to improve and strengthen the zoonotic disease surveillance system in India.

Seroprevalence and risk factors for brucellosis in small ruminant flocks in Karnataka in the Southern Province of India.

BACKGROUND AND AIM: Brucellosis is a zoonotic disease of high economic and public health importance in large and small ruminant populations worldwide. A cross-sectional study was conducted to determine the seroprevalence and risk factors of brucellosis in small ruminants in organized farms in the southern region of India. MATERIALS AND METHODS: Farms exclusively rearing sheep and goats were selected based on the number of animals (small, medium, or large) and the location of the farm (urban, periurban, or rural). A total of 1499 serum samples; 1001 from sheeps and 498 from goats were sourced from six sheep and four goat farms and tested using Rose Bengal Plate and indirect Enzyme-Linked Immunosorbent Assay tests. RESULTS: The apparent prevalence of brucellosis was higher in sheep (8.29%, 95% CI 6.7-10.1) than goats (5.82%, 95% CI 4.0-8.2). The true adjusted population level seroprevalence was also higher in sheep, at 7.7% (95% CI 6.0-9.6) than in goats, at 5.1% (95% CI 3.2-7.6). According to bivariate categorical analysis, six highly significant (p<0.001) animal- and farm-level risk factors for sheep were age, breed, number of lambings, history of abortion, rural farms, and presence of dogs on the farm. In goats, five significant risk factors were found: History of abortion, separate sheds, dogs on the farm, weekly veterinary consultation, and lack of brucellosis awareness. In a logistic regression model, abortion (OR adjusted 10.8, 95% CI 1.2-96.12), rural farms (OR adjusted 8.5, 95% CI 3.6-20.0), and absence of separate sheds on the farms (OR 1.9, 95% CI 1.1-3.5) were found to be significant risk factors for ovine brucellosis. CONCLUSION: The use of complementary measures to tackle the multiple animal- and farm-level risk factors may help to reduce the disease burden in the absence of a vaccination policy for small ruminants in India.

Spatial sero-prevalence of brucellosis in small ruminants of India: Nationwide cross-sectional study for the year 2017-2018.

Brucellosis in small ruminants caused mainly due to Brucella melitensis is an important zoonotic disease characterized by abortion, retained placenta, infertility, orchitis, epididymitis and rarely arthritis. Small ruminants are the main source of economy for the rural and marginally poor farmers and brucellosis is resulting in huge economic losses due to abortions and infertility and causing public health concern among the small ruminant keepers. Bovine brucellosis control programme has been implemented in India and small ruminants are left out of the programme mainly due to paucity of brucellosis status. The present cross-sectional study based on stratified random sampling was undertaken during 2017-18 to provide the nationwide brucellosis sero-prevalence in small ruminants. A total of 24,056 small ruminant serum samples (sheep samples = 8,103 [male-2,440 and female-5,663] and goat samples = 15,953 [male-4,331 and female-11,622]) sourced from 27 out of 29 states and two out of seven union territories (UTs), 350 districts of total 640 districts (54.68% of the Indian districts) and from 1,462 villages out of 6,40,867 villages (43.83% of the Indian villages). The serum samples were tested by indirect ELISA and overall brucellosis apparent and true prevalence of 7.45 (95% CI: 7.13-7.79) and 3.79 (95% CI: 3.44-4.17) was recorded. Significantly higher brucellosis sero-prevalence (p < .0001) was observed in sheep (11.55%) than goats (5.37%). Similarly, brucellosis seropositivity was highly significant in females compared to males in both sheep and goats. Countrywide, greater than 5% brucellosis sero-prevalence in sheep and goats was recorded in 14 and 10 states, respectively, indicating endemicity of the disease. The study provided the latest update on nationwide spatial sero-prevalence of small ruminant brucellosis which will aid government to strengthen regular surveillance and vaccination to reduce the disease burden and public health problems in the country.

Knowledge and practices of dairy farmers relating to brucellosis in urban, peri-urban and rural areas of Assam and Bihar, India.

BACKGROUND: Brucellosis is one of the most common zoonotic diseases in the world. This study aimed at assessing farmers' knowledge about brucellosis as well as practices relevant to transmission of brucellosis and their associated determinants. RESULTS: Few farmers knew about brucellosis (3.4%, n = 18) and its zoonotic importance (0.8%, n = 4). Knowledge about brucellosis was higher for farmers with a larger herd size (p < 0.001) and fully using a stall-fed system (p < 0.001). Training on dairy cattle management (p < 0.001), training on animal disease (p < 0.01), consultation with veterinarians (p < 0.001) and farms being in urban areas (p < 0.01) were also significantly positively associated with knowledge about brucellosis. No significant association was observed between farmers' knowledge about brucellosis and state, family size, education, age or gender of the farmers. Farmers knowledge about brucellosis was significantly associated with certain practices that include use of disinfectant while cleaning farms (p < 0.05), animal movement (p < 0.01), introduction of new animals (p < 0.05) and raw milk consumption (p < 0.05). The study did not find any association between knowledge about brucellosis and method of disposal of aborted materials, personal hygiene and quarantine practices. CONCLUSION: More interaction with veterinarians and training on animal management may be an important tool for generating awareness among the farming community for reducing transmission of the disease.

Prevalence and antibiotic susceptibility of Mannheimia haemolytica and Pasteurella multocida isolated from ovine respiratory infection: A study from Karnataka, Southern India.

BACKGROUND AND AIM: Respiratory infection due to Mannheimia haemolytica and Pasteurella multocida are responsible for huge economic losses in livestock sector globally and it is poorly understood in ovine population. The study aimed to investigate and characterize M. haemolytica and P. multocida from infected and healthy sheep to rule out the involvement of these bacteria in the disease. MATERIALS AND METHODS: A total of 374 healthy and infected sheep samples were processed for isolation, direct detection by multiplex PCR (mPCR), and antibiotic susceptibility testing by phenotypic and genotypic methods. RESULTS: Overall, 55 Pasteurella isolates (27 [7.2%] M. haemolytica and 28 [7.4%] P. multocida) were recovered and identified by bacteriological tests and species-specific PCR assays. Significant correlation between the detection of M. haemolytica (66.6%) with disease condition and P. multocida (19.1%) exclusively from infected sheep was recorded by mPCR. In vitro antibiotic susceptibility testing of 55 isolates revealed higher multidrug resistance in M. haemolytica (25.9%) than P. multocida (7.1%) isolates. Descending resistance towards penicillin (63.6%), oxytetracycline (23.6%), streptomycin (14.5%), and gentamicin (12.7%) and absolute sensitivity towards chloramphenicol were observed in both the pathogens. The antibiotic resistance genes such as strA (32.7%) and sul2 (32.7%) associated with streptomycin and sulfonamide resistance, respectively, were detected in the isolates. CONCLUSION: The study revealed the significant involvement of M. haemolytica together with P. multocida in ovine respiratory infection and is probably responsible for frequent disease outbreaks even after vaccination against hemorrhagic septicemia in sheep population of Karnataka, southern province of India.