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Tick-borne pathogens pose a significant global threat, causing substantial economic losses to the dairy industry. In India, tropical theileriosis, anaplasmosis, babesiosis, and trypanosomiasis are major hemo-parasitic diseases affecting bovines. A cross-sectional study was conducted to determine the prevalence of hemo-parasites in different farms in India. PCR assays were employed to detect carrier status, using gene targets msp1b, tams1, rap-1, ama1, and ITS1 for A. marginale, T. annulata, B. bovis, B. bigemina, and Trypanosoma species, respectively. Out of the 578 apparently healthy animals screened, 30.45% (95% CI: 26.84-34.32%) were infected with at least one hemo-parasite. Cattle showed an overall positivity of 32.87%, while buffaloes had a prevalence of 15.19%, which was statistically significant ( p  < 0.001). Interestingly, prevalence was higher in indigenous cattle (47.81%) compared to cross-breeds (25.53%) and exotics (14.62%), with a statistically significant difference ( p  < 0.001). The prevalence of hemo-parasites varied widely among the farms, ranging from 5.77 to 100%. A. marginale was the most prevalent parasite (23.70% of animals), followed by T. annulata (13.67%), Babesia species (1.90%), and Trypanosoma species (1.56%). Enzootic instability was observed in six of the eight farms, indicating a potential for future outbreaks. Co-infection was detected in 60 out of 176 animals positive for hemo-parasites, with 59 animals co-infected with A. marginale and T. annulata , and only one cross-breed cattle infected with both Anaplasma marginale and Babesia bigemina . The findings highlight the prevalence of hemo-parasites in farms, underscoring the need for whole-herd screening, treatment of infected animals, and improvement in farm management practices to prevent production losses caused by these pathogens., Supplementary Information: The online version contains supplementary material available at 10.1007/s12639-024-01673-3., Competing Interests: Conflict of interestThe authors declare no conflict of interest related to this article., (© Indian Society for Parasitology 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Bovine alphaherpesvirus-1 (BoAHV-1) is an important viral pathogen that causes significant economic losses to the dairy industry. The present study aimed to determine the prevalence of BoAHV-1 in cases of bovine reproductive disorder. Clinical samples were collected from various villages in Gujarat using specialized FTA ® cards and were tested using real-time PCR assay targeting the gB gene of BoAHV-1. Out of 401 animals, 18.20% (95% CI: 14.74-22.28%) tested positive for BoAHV-1 DNA. The percentage positivity of BoAHV-1 was 20.37% in abortion cases and 19.55% in retention of fetal membrane cases, while only one out of nine metritis cases screened in the study was positive for BoAHV-1 DNA. A higher percentage positivity in buffaloes (22.14%) compared to cattle (16.30%) was recorded, but this difference was not statistically significant ( p  = 0.169). The frequency of BoAHV-1 detection was higher among crossbreeds (16.76%) and exotics (19.61%) than among indigenous cattle (8.82%), although this difference was not statistically significant ( p  = 0.400). There was also no significant difference in frequency distribution among animals of varying parity, ranging from 15.20 to 33.33% ( p  = 0.540). This study confirms the widespread circulation of BoAHV-1 and highlights the need for its control and prevention., Competing Interests: Conflict of interestThe authors declare no conflict of interest related to this article., (© The Author(s), under exclusive licence to Indian Virological Society 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Brucella melitensis primarily affects sheep, goats and is associated with brucellosis in humans, which is one of the world's most widespread neglected zoonotic disease. The current study attempted the determination of genetic diversity through comparative genome analysis of B. melitensis strains reported from India with other countries. The study also reports the isolation and identification of B. melitensis BMNDDB8664 from a cow with a history of abortion, whole-genome sequencing (WGS), determination of virulence factors, genotyping, and comparative genome analysis. Multilocus sequence typing, Multiple locus variable number of tandem repeats analysis (MLVA), and WGS based phylogeny revealed the predominance of ST-8 and genotypes (116 and II respectively) that clustered to the East Mediterranean lineage. Identification of hitherto unreported genotypes by MLVA also indicated the existence and circulation of West Mediterranean and American lineages in India. Though the AMOS-PCR results suggest the BMNDDB8664 isolate as Brucella abortus , the outcomes from multiplex PCR, ribosomal multilocus sequence typing, and WGS analysis confirmed it as B. melitensis . The analysis revealed the presence of adeF gene (aids conferring resistance to fluoro-quinolone and tetracyclines). The isolate lacked two important T4SS genes virB2 and virB7 genes (roles in infection and rifampicin resistance respectively) and also lacked the Brucella suis mprF gene that aids intracellular survival. Further, BMNDDB8664 lacked some of the genes associated with LPS synthesis ( wbkB , wbkC ) and transport ( wzm , wzt ) and hence, is most likely a rough strain. WGS-based phylogenetic analysis revealed close genetic relatedness of this BMNDDB8664 with a sheep isolate and two human isolates. The results prompt systematic, broad-based epidemiological studies on brucella infection at the species level. For effective control of human brucellosis, a concerted One Health approach with studies encircling the identification of aetiology at species, strain level to find their prevalence, spread, and inter-host transmission patterns need to be understood, for better design and implementation of effective control strategies in India and other endemic regions., Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-023-01081-w., Competing Interests: Conflict of interestNone., (© Association of Microbiologists of India 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Background & Objectives: Crimean-Congo hemorrhagic fever (CCHF) is an emerging tick-borne viral zoonotic disease of public health importance. Cattle and buffaloes although not showing any clinical symptoms, can be infected by the CCHF virus and act as sources of infection to human beings. The prevalence of CCHF in cattle and buffaloes is important from One health perspective for control of CCHF in humans., Methods: A cross-sectional study was undertaken to ascertain the prevalence of CCHFV in cattle and buffaloes of India., Materials and Methods: A total of 804 serum samples from four states of India (Gujarat and Rajasthan: human outbreaks reported; Punjab and Haryana: no outbreak reported) were screened by ELISA test detecting nucleoprotein antibodies of CCHFV., Results: The overall true prevalence was 8.63% (95% CI: 6.76% - 10.9%). The highest prevalence was recorded in Rajasthan (13.24%) followed by Gujarat (8.68%), Haryana (6.84%), and Punjab (6.51%). Prevalence of CCHF was higher in cattle (9.92%) than buffaloes (5.84%); in females (10.87%) than males (4.99%); in adults (10.18%) than young ones (5.66%). Interestingly, higher seropositivity was recorded in indigenous cattle (12.04%) than in exotic and cross-breed cattle (1.69%) which was statistically significant (p=0.001)., Interpretation & Conclusion: These findings revealed CCHF virus is circulating unnoticed and the prevalence has increased over time which is of public health concern., Competing Interests: None

Bovine tropical theileriosis, a tick-borne disease, causes huge economic loss to the Indian dairy industry. Theileriosis in India is mainly caused by Theileria annulata, although the presence of T. orientalis has also been reported. The present study was undertaken to investigate the deaths of cross-bred Holstein Friesen (CBHF) cows on a farm in the state of Telangana, India. Deceased animals had recently calved and prior to death had developed high fever (107 °F) and anaemia. Infected cows were infested with ticks (Hyalomma species). Theileria piroplasms were noticed in the Giemsa stained blood smears. PCR assays further confirmed the presence of Theileria in the blood samples of the infected cows. Partial Tams1 gene sequences from the infected animals shared 99.87% to 100% identity scores with the sequences of Sri Lankan isolates recently proposed as a novel Theileria species (provisionally designated as Theileria sp. Yokoyama). To the best of our knowledge, this is the first report of the novel species of Theileria from India. Infected animals were effectively treated with buparvaquone and oxytetracycline. The introduction of new animals into the farm without risk assessment was found to be a major cause of the outbreak., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Seven ELISA kits were evaluated for the fitness of purpose in diagnosing brucellosis among cattle and buffaloes in the endemic scenarios of India. The sera (675 numbers) for the study were sourced from brucellosis-free as well as infected herds. The diagnostic sensitivity (dsn) and specificity (dsp) of the kits were determined by three approaches: based on the results of the Rose Bengal test, history of the animals (sera from infected or naïve animals), and based on the results obtained from the 'majority of the tests'. The dsn and dsp ranged from 65.10% to 98.66%, and 98.04% to 100% respectively. The results and suitability of the kits for diagnostic application in various epidemiological situations were discussed., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Information on the prevalence of infectious agents in dairy farms forms the basis for formulating a suitable control strategy; especially in endemic situations., Aims: A cross-sectional study was undertaken to determine the prevalence of six economically important bovine diseases, causing reproductive disorders including bovine abortion in organized dairy herds in India., Methods: A total of 1,075 animals (cattle and buffaloes) from 09 dairy farms were screened by ELISA tests., Results: Bovine viral diarrhoea (BVD) was the most prevalent (56.5%) disease followed by infectious bovine rhinotracheitis (IBR) (45.4%). Prevalence of Q-fever (5.4%) and neosporosis (6.1%) were less on the farms. Although 16.3% of the samples turned positive for brucellosis, the contribution of calf-hood vaccination ( B. abortus S19 vaccine) to the prevalence of antibodies cannot be ruled out. The overall prevalence of bovine anaplasmosis, known to cause sporadic abortions in dairy herds, was 34.1% in the 9 farms with a prevalence of less than 20% in 5 farms. Infection of multiple abortifacient (seroprevalence to more than two pathogens) was recorded in 56.8% of animals. A very strong association was observed between BVD and brucellosis (Odds ratio 14.2; P<0.001). Further, a positive association was also seen between seroprevalence of IBR and anaplasmosis, and neosporosis and Q fever (P<0.05)., Conclusion: Viral diseases were found to be more common in the dairy herds than bacterial and protozoan diseases. Increased susceptibility of IBR seropositive cows to other bacterial and viral infections was observed., Competing Interests: The authors declare that there are no conflicts of interest in this article.

Bovine alphaherpesvirus-1 (BoHV-1), the causative agent of infectious bovine rhinotracheitis (IBR), is an economically important viral pathogen affecting cattle and buffaloes. Serological assays are mostly used for detection of the antibodies, but variation has been detected in the diagnostic performances of the individual assay. In the present study, four commercially available ELISA kits {two indirect ELISA (kits A and B) and two blocking ELISA (kits C and D)} were evaluated for the detection of antibodies against BoHV-1 in Indian cattle and buffaloes (fitness of purpose). The diagnostic sensitivity (dsn) and specificity (dsp) of these kits were determined by three ways; considering virus neutralization test (VNT) as gold standard test, using pre-test information of the samples, and majority of tests. Screening of 200 known negative sera (124 cattle, 76 buffaloes) sourced from IBR free farms revealed gB based ELISA kits are more specific than the indirect ELISA kits. Testing of 125 known positive sera (81 cattle, 44 buffaloes) suggests kit B be most sensitive followed by kit C, A and D. Interestingly, kit D was found to be most sensitive for detection of vaccination-induced BoHV-1 antibodies followed by kit B. Similar trend were also observed in the limit of dilution experiment performed using known infected and vaccinated sera. VNT was found to be the most specific test and its use as the gold standard test revealed all kits to have more than 99 % sensitivity. All the ELISA kits could detect BoHV-1 specific antibodies in the IBR vaccinated calves as early as 11 days post-vaccination. In Kappa statistics, an almost perfect agreement between the ELISA kits was recorded. The overall performance of the kits in serodiagnosis of IBR as determined by the area under curve in ROC analysis was good., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Bovine anaplasmosis is one of the most important tick borne disease in ruminants causing huge economic loss to the dairy industry. A cross-sectional study was carried out to detect serum antibodies to Anaplasma infection in cattle and buffaloes housed in 14 organized herds located at various climatic zones spreading over 9 different states in India. A total of 911 serum samples, collected from 667 cattle and 244 buffaloes, were subjected to a competitive enzyme linked immune-sorbent assay detecting an epitope of major surface protein 5 (MSP5) of Anaplasma . The overall true prevalence was 48.72% (95% CI 45.13-52.32%). The prevalence rate was higher in cattle (51.58%) than buffaloes (40.89%) and the difference was statistically significant ( p  < 0.05). Indigenous cattle (59.30%) showed higher seropositivity than crossbreed (57.16%) and exotic cattle breeds (42.28%). Although statistically not significant, female (52.37%) showed higher seropositivity than male (46.43%). Similarly, significant difference in prevalence ( p  < 0.05) was observed for animals reared in different climatic zones with highest prevalence recorded in arid zone (90.49%) and lowest in semi-arid zone (29.83%). Very wide variation in prevalence (9.95-100%) was recorded between farms. The present study indicates endemicity of Anaplasma in India, similar to other tropical and sub-tropical countries of the world. Endemic instability was recorded in some of the studied farms suggesting possibility of outbreak of new clinical cases resulting in economic loss. Therefore, suitable policies and procedures for prevention and control of Anaplasma infection should be adopted in these farms., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Indian Society for Parasitology 2020.)

Abortions in dairy animals can be caused by several infectious agents. Identification of the actual causal agent(s) is important for formulating suitable control strategies. A 3-year (2016-2018) longitudinal study was conducted in a dairy farm following an abortion storm in the mid- to late gestations. The investigation focused on the seven major infectious abortifacient in cattle, viz. bovine alphaherpesvirus-1 (BoHV-1), bovine viral diarrhoea virus (BVDV), Neospora caninum, Brucella abortus, Coxiella burnetii, Leptospira Hardjo, and Listeria monocytogenes. High seroprevalence was observed for BVDV (79.4%), Leptospira (70.5%), BoHV-1 (53.5%), and Brucella (45.0%) at the beginning of the investigation (August 2016). The incidence proportion increased for BVDV, Leptospira, and Brucella in the following years of the investigation. A strong association of Brucella seropositivity with history of abortion (OR = 3.27) was recorded. Incidence of BoHV-1 reduced during the period of study coincident with systematic IBR inactivated marker vaccination of the herd. Sixty-four abortion cases were investigated for the identification of causative agent(s) by microbial culture, serological (ELISA), and molecular detection (PCR/ real-time PCR). Antibodies to BVDV, Brucella, BoHV-1, Leptospira, Neospora, and Coxiella were detected in 63, 61, 56, 35, 5, and 6 aborting cattle, respectively. Real-time PCR/PCR of clinical specimens detected DNA of Brucella, BoHV-1, Coxiella, Leptospira, and Listeria in 34, 13, 12, 9, and 4 abortion cases, respectively. BVDV and Neospora were not detected in any specimen samples. Brucella abortus isolated from the farm was determined as ST1 by multi-locus sequence typing (MLST). DNA of multiple agents were detected in 21 of the 64 cases (43.75%). Overall, the data suggests, Brucella was the major causative agent, although multiple causative agents circulated in the farm.

Infectious bovine rhinotracheitis (IBR) caused by bovine alphaherpesvirus 1 (BoHV-1) is an economically important disease of cattle and buffaloes. Following acute infection, the virus usually attains latency in the sensory neurons. Stress-induced reactivation of latency can cause the infected animals to intermittently shed the virus in body secretions including semen. A longitudinal analysis was carried out to study BoHV-1 shedding in the semen of IBR seropositive cattle and buffaloes. The study involved data generated from the screening of 119,850 extended frozen semen (EFS) batches, collected from 1,229 IBR seropositive bulls, over a period of four years (April 2015 to March 2019). A TaqMan based real-time PCR assay was employed to detect the gB gene BoHV-1 DNA in the EFS batch samples. Each sample was tested in duplicate and amplification in any of the replicates at or below the threshold cycle (Ct ≤ 40) was considered positive. The overall positivity of BoHV-1 in EFS batches was 1.18%. About 41% of the bulls (509 of 1,229) were found to have excreted the virus in semen at least once during the study period. The frequency of viral shedding in buffaloes (0.96%) was significantly lower than that of cattle (1.3%) (p < 0.001). No significant difference was noted in the rate of shedding between the first and the second ejaculates collected on the same day (p = 0.607). The rate of shedding also did not vary among various breeds of cattle (p = 0.454) or with the age of the bulls (p = 0.054). No significant variation in the shedding rate was observed in cattle across different seasons (p = 0.101); while in buffaloes, the rate was higher in autumn (1.2%) than in winter (0.7%) (p = 0.037). The difference in positivity among semen stations was statistically significant (p < 0.001). Analysis of data revealed that ≥100 EFS batch samples/bull were screened from 361 of the 1,229 bulls included in the study. None of the EFS batches screened from 39 of these 361 bulls were found positive during the four years, suggesting they were non-shedders. Further research is warranted to delineate the underlying features of the seropositive non-shedders; following which an adequate risk assessment may be made for the maintenance of infected but non-shedding bulls in semen production., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest to this article., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The extended frozen semen (EFS) batches produced from infectious bovine rhinotracheitis (IBR) sero-positive cattle and buffalo bulls housed in various semen stations in India are transported to the testing laboratory in liquid nitrogen (LN 2 ) for screening bovine herpesvirus-1 (BoHV-1). This procedure is laborious and poses LN 2 related hazards. An alternative logistics for transportation of samples was investigated. Use of Flinders Technology Associates (FTA ® ) elute card was evaluated for transportation of extended bovine semen to screen BoHV-1 DNA by real-time PCR targeting gB gene and the method was compared with the OIE approved Chelex resin based method. A protocol for extraction of BoHV-1 DNA from FTA ® card spotted with extended semen was optimized. The viral DNA was found to be stable on FTA ® card for at least 28 days when the cards are stored at 4°-37 °C. The analytical sensitivity for the assay was determined using variable dilutions of BoHV-1 spiked semen and positive plasmid harbouring gB gene (97bp) spotted onto FTA ® card and it was found to be 10 0.8 TCID 50 /ml or 100 copies respectively in real-time PCR. The test could detect as low as 10 0.008 TCID 50 /ml or 1 copy of positive plasmid when more number of replicates (n = 6) of the same sample were tested. This sensitivity was found to be comparable to Chelex method and both the methods demonstrated a very strong correlation (r = 0.9774; 95% CI: 0.9620-0.9860) in terms of Ct value (p < 0.0001). The diagnostic sensitivity and specificity of the FTA method in comparison to the Chelex method was 83.08% (95% CI: 71.73%-91.24%) and 93.23% (95% CI: 89.38%-96.01%) respectively when 316 samples were screened by both the methods. The degree of agreement between these two tests was good (Kappa value: 0.738; 95% CI: 0.646-0.829). The method was found to be robust and highly repeatable in inter-assay and intra-assay precision testing. The result suggests that the FTA ® card holds promise as an alternative system for transportation of EFS for downstream screening of BoHV-1 DNA., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Foot-and-mouth disease (FMD) is a highly contagious and rapidly transmissible disease of cloven footed animals. Emergence of genetically divergent strains of FMD virus (FMDV) is a major concern globally. FMD is endemic in India and three serotypes (O, A and Asia 1) prevail. The study was undertaken to characterize the isolates from the state of Odisha, India both genetically and antigenically. FMDV was detected in 7 of the 17 clinical samples collected from FMD affected/suspected animals, in which serotype O and A were found in three and four samples, respectively. Serotype O field isolates clustered in an unnamed group (designated here as Eastern cluster) circulating mostly in the Eastern region of the country and had 10-12.7% divergence from the Ind2001 lineage circulating predominantly throughout the country. The serotype A isolates sequenced in this study was grouped within VP3 59 -deletion group of genotype 18, precisely in clade 18c, having high genetic homology to the virus circulating in the neighboring states, suggesting interstate movement. Both the serotype O and A isolates showed good antigenic relationship value with the respective vaccine strains currently used in the country.

Multilocus sequence typing (MLST), a sequence-based typing method for bacterial pathogens, is currently the best method for long-term epidemiological study and to understand the population structure of the bacteria. This investigation was carried out to study the diversity of Pasteurella multocida isolates circulating in India. Ten different sequence types (ST) identified in this study are ST 122 from cattle, goat, mithun and pig; ST 50 from pig; ST 9 from cattle and sheep; ST 229 from cattle and goat; ST 71 and ST 277 from cattle; and ST 129, ST 280, ST 281 and ST 282 from avian species. Of these, ST 277, ST 280, ST 281 and ST 282 were identified for the first time. The analysis of results provides novel epidemiological information on the circulation of multiple STs across India. The majority of STs or their variants identified in this study have already been reported from different parts of the globe. This suggests that probably transboundary spread of strains across countries and continents has occurred across evolutionary time and is still happening. The isolation of ST 122 from small ruminants and pigs suggests that these species may be included in the preventive vaccination policy for effective control of haemorrhagic septicaemia in India., (© 2014 Blackwell Verlag GmbH.)

Aim: The aim of the present study was to carry out antigenic detection of bluetongue virus (BTV) among the small ruminant population of two different geographical regions of Odisha (coastal and central) using recombinant VP7 (r-VP-7) based sandwich enzyme-linked immunosorbent assay (s-ELISA)., Materials and Methods: Blood samples (n=274) were collected from two different geographical pockets of Odisha, which covered mostly the coastal and central regions. Of the total samples under study 185 were from goat and 89 were from sheep. The blood samples were tested for the presence of BTV antigen by r-VP7 based s-ELISA., Results: r-VP-7 s-ELISA detected BTV antigen in 52.43% and 44.94% of the goat and sheep population under study, respectively. This study highlights the antigenic persistence of BTV in the state for the 1(st) time., Conclusion: This high antigenic presence in both sheep and goat population suggests an alarming BTV infection in field conditions which warrants more systematic study directed toward isolation and characterization studies as well as the implementation of control strategy for BT in Odisha.

RNA virus population exists as a complex distribution of non-identical but closely related sequences known as viral quasispecies. Variant strains are selected from this quasispecies population in response to changing environment. The quasispecies dynamics of a virus existing within an infected host differs from that in a cell culture-adapted population. This study was carried out to explore the genetic variations present in the VP1 coding region of the foot-and-mouth disease (FMD) virus serotype O derived directly from infected cattle tongue epithelium. Molecular clonal populations of two serotype O strains belonging to lineages Ind2001 (IND 30/2011) and PanAsia2 (IND 5/2011) were sequenced at VP1 coding region. For IND 30/2011, 19 clones were sequenced and analysis showed variations at 12 nucleotide positions (nt) resulting in 8 amino acid (aa) replacements. Similarly, for IND 5/2011 virus, 18 clones were sequenced, of which six showed nt variations leading to 3 aa replacements. Most of the variable positions mapped to the surface-exposed loops and some of them were found in the neutralizing antigenic sites (position 81, 149, 169, 186 and 202 of IND 30/2011 and 141 of IND 5/2011), which potentially could be beneficial in rapid adaptive evolution of the virus by giving rise to antigenic variants to overcome neutralizing antibodies. These findings encourage further research into the landscape of the viral quasispecies population in vivo and its implication for viral ecology.

Pasteurellosis in small ruminants affects the livelihood of small and marginal farmers of India. The present study was undertaken to understand the trends in gene carriage and antibiotic resistance pattern of Pasteurella multocida isolates recovered from small ruminants over a period of 10 years in India. A total of 88 P. multocida isolates of small ruminant origin were subjected to virulence gene profiling for 19 genes by PCR and antibiogram study employing 17 different antibiotics. Virulence genes like exbB, exbD, tonB, oma87, sodA, sodC, nanB and plpB (100% prevalence) and ptfA and hsf-2 (>90% prevalence) were found to be uniformly distributed among isolates. Unexpectedly, a very high prevalence (95.45%) of pfhA gene was observed in the present study. Dermonecrotoxin gene (toxA) was observed in 48.9% of isolates with highest occurrence among serotype A isolates and interestingly, one of each isolate of serotype B and F were found to carry this gene. Antimicrobial susceptibility testing revealed 17.04% isolates to be multidrug resistant. Amongst all the antibiotics tested, most of the P. multocida isolates were found to be susceptible to enrofloxacin and chloramphenicol. This study highlights novel epidemiological information on frequency and occurrence of virulence genes among Indian isolates from small ruminants., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In this study, 108 P. multocida isolates recovered from various host animals such as cattle, buffalo, swine, poultry (chicken, duck, and emu) and rabbits were screened for carriage of 8 virulence associated genes. The results revealed some unique information on the prevalence of virulence associated genes among Indian isolates. With the exception of toxA gene, all other virulence associated genes were found to be regularly distributed among host species. Association study between capsule type and virulence genes suggested that pfhA, nanB, and nanH genes were regularly distributed among all serotypes with the exception of CapD, whereas toxA gene was found to be positively associated with CapD and CapA. The frequency of hgbA and nanH genes among swine isolates of Indian origin was found to be less in comparison to its equivalents around the globe. Interestingly, very high prevalence of tbpA gene was observed among poultry, swine, and rabbit isolates. Likewise, very high prevalence of pfhA gene (95.3%) was observed among Indian isolates, irrespective of host species origin.

Identification of outer membrane proteins (OMPs) is important to understand the bacteria structure and function, host-pathogen interaction, development of novel vaccine candidates, and diagnostic antigens. But till now the key antigens of P. multocida B:2 isolate causing haemorrhagic septicaemia (HS) in animals are not clearly defined. In this study, P52 strain of P. multocida serotype B:2 was grown in vitro under iron-rich and iron-limited condition. The OMPs were extracted by sarkosyl method followed by SDS-PAGE and the proteins were identified by MALDI-TOF/MS analysis. In total, 22 proteins were identified, of which 7 were observed exclusively under iron-limited condition. Most of the high molecular weight proteins (TbpA, HgbA, HgbB, HasR, IroA, and HemR) identified in this study were involved in iron acquisition. Some hypothetical proteins (HP-KCU-10206, HP and AAUPMB 08244, HP AAUPMB 21592, HP AAUPMB 19766, AAUPMB 11295) were observed for the first time in this study which could be unique to serotype B:2. Further functional in vivo study of the proteins identified are required to explore the utility of these proteins in developing diagnostics and vaccine against HS.

Foot-and-mouth disease virus (FMDV) is constantly evolving under neutralizing antibody pressure in either naturally infected or vaccinated animals. This study was carried out to understand the dynamics of evolution of antigenic sites. Neutralizing antibody-resistant populations of three strains of FMDV serotype O (INDR2/1975, IND120/2002 and IND271/2001) were isolated by serial propagation in BHK-21 cells in the presence of sub-neutralizing level of bovine vaccinal sera (BVS). In the partial neutralization escape variants, fixation of aa substitutions were observed at critical residues of all established antigenic sites of serotype O {144 of VP1 (site 1), 45 and 48 of VP1 (site 3), 72 and 134 of VP2 (Site 2)} except site 4 and 5. In majority of the variant populations, site 3 was found to be substituted and therefore immunodominance may not be associated with a particular site, rather it appears to be a virus strain and infected host specific affair. Substitutions were also observed in proximity to the identified residues {41 and 51 (βB-βC loop), 133, 140 and 143 (βG-βH loop), 201, 204 and 209 (C terminus) of VP1, 71 and 75 (βB-βC loop), 131 (βE-αB region), 174 and 179 (βG-βH loop) and 219 (C terminus) of VP3} within antigenic sites of serotype O or other serotypes which could be significant in terms of neutralizing antibody binding and immune escape. Presence of similar residues in the Indian field viruses as selected in the variants supports the importance of these sites in antigenic diversification of serotype O FMD virus., (Copyright © 2013 Elsevier B.V. All rights reserved.)

India is endemic for foot-and-mouth disease (FMD), and goats constitute the second largest susceptible population of domestic livestock. FMD surveillance and control strategies in the country largely ignore small ruminants, known to be critical in the epidemiology of the disease. Here, serological investigations were carried out to generate estimates of antibody prevalence in goats of Orissa state to both non-structural (NSP-Ab) and structural proteins (SP-Ab) of FMD. The apparent overall NSP-Ab and SP-Ab seroprevalences were 38% and 20.7%, respectively, which signifies a very high level of FMD virus circulation in the goat population despite the lack of clinical signs in this species. The apparent prevalence of NSP-Ab and SP-Ab was positively correlated in the sampling areas. Interestingly, the values found for NSP-Ab prevalence were almost consistently higher than those found for SP-Ab prevalence. This could have been attributable to either issues related to sensitivity and specificity of the test systems employed or differences in the post-infection kinetics of NSP- and SP-Ab. The pattern that emerged from SP-Ab analysis indicated goats being infected with all three prevalent serotypes (O, A and Asia 1) and reinforces the concept that non-vaccinated goats can be exploited as tracer animals for detecting serotypes involved in outbreaks. The results underscore the requirement to bring caprine species under comprehensive surveillance and vaccination campaigns to check silent amplification, excretion and transmission of the virus., (© 2010 Blackwell Verlag GmbH.)