Malignant catarrhal fever (MCF) presents a sporadic yet significant threat to livestock and wildlife. A comprehensive investigation in Karnataka, India into the prevalence and transmission patterns of sheep-associated MCF (SA-MCF) was conducted. A total of 507 sheep peripheral blood leukocyte samples from 13 districts along with 27 cows and 10 buffalo samples from various regions in Karnataka were tested for SA-MCF infection i.e. Ovine gammaherpesvirus 2 (OvHV-2) using heminested PCR. Furthermore, serum samples collected from 73 cows and 15 buffalo suspected of MCF were tested using a commercially available ELISA kit. Additionally, histopathological examinations of affected tissues and phylogenetic analysis of viral tegument protein sequences were conducted. Our findings indicated a 20.11%, 33.33% and 20% positivity for OvHV-2 in sheep, cows and buffalo respectively by PCR. Statistical analysis revealed a significant association between the age of sheep and the detection of OvHV-2. Seven cows and one buffalo serum samples tested positive for ELISA. Clinical findings in bovids were consistent with typical MCF signs, and histopathological results revealed multi-organ involvement characterised by necrotising vasculitis and lymphoid hyperplasia. The nucleotide pairwise identity matrix revealed 99.5% identity between the sequences obtained in the study with sequences from other states. The phylogenetic analysis of partial tegument protein sequences from bovid and sheep samples suggested a close genetic relationship between the local OvHV-2 strains and those from various global regions. Crucially, this study underscores the widespread presence of SA-MCF in Karnataka, with significant implications for both livestock management and wildlife conservation.

African Swine Fever Virus (ASFV) poses a significant threat to global swine populations, with devastating economic and agricultural implications. This review article provides a comprehensive examination of various facets of ASFV, encompassing its structure, entry mechanism, transmission dynamics, clinical signs, pathogenesis, diagnosis and control strategies. The complex virion architecture, including the multilayered core and distinctive outer envelope, is explored, shedding light on key elements influencing the virus’s stability and infectivity. The intricate mechanisms governing ASFV entry into host cells are discussed, emphasizing the interplay between viral proteins and cellular receptors. Insight into the virus-host interaction provides a foundation for understanding the initial stages of infection, influencing subsequent pathogenesis. Transmission dynamics, a critical aspect of ASFV epidemiology, are examined, encompassing both direct and indirect modes of spread. Factors influencing the persistence of ASFV in diverse environments and the role of vectors in disease dissemination are explored to elucidate the complex transmission pathways. Clinical signs and pathogenesis of ASFV infection are thoroughly reviewed, outlining the diverse manifestations in swine species. The immunopathological responses and host factors influencing disease severity are discussed, enhancing our understanding of ASFV pathobiology. A comprehensive understanding of diagnostic tools is pivotal for timely and accurate disease detection, enabling swift intervention measures. In conclusion, this review provides a nuanced and integrative overview of ASFV, offering valuable insights for researchers, veterinarians and policymakers engaged in combatting this significant threat to the swine industry.

Evaluation of the effectiveness of vaccination of animals against rabies is not routinely implemented. In cases where it is carried out, the rapid fluorescent focus inhibition test (RFFIT) or the fluorescent antibody virus neutralization (FAVN) test are the recommended tests. However, both of these tests require handling of live rabies virus (RABV), and are cumbersome to perform. In view of this, the enzyme-linked immunosorbent assay (ELISA) has been proposed as a surrogate test; however, availability of appropriate antigen is a major impediment for the development of ELISAs to detect anti-rabies antibodies. The most widely used antigen is the RABV glycoprotein (G) purified from cell culture-propagated virus, which requires a biosafety level 3 containment. The alternative is to use recombinantly expressed G, which needs to be to be properly glycosylated and folded to serve as the best antigen. The most suitable system for its production is the baculovirus expression system (BVES). However, purification of RABV G is challenging. We therefore tested partially purified preparations in the form of extracts of insect cells infected with baculovirus expressing RABV G, against sera from vaccinated dogs in an indirect ELISA. The results showed good concordance against RFFIT, with sensitivity and specificity of 90.48% and 80.00%, respectively. The system may be used for quick screening to determine the presence and an approximate level of antibodies, and can be modified to enable monitoring of mass dog vaccination programs, as well as to facilitate certification of dogs intended for international travel and transportation.

The present study is focused to express the recombinant Leptospira surface adhesion-44 (rLsa44) protein of pathogenic Leptospira interrogans serovar Pomona in Escherichia coli and its diagnostic application in the latex agglutination test (LAT) system to detect anti-leptospiral antibodies in the bovine sera. The Lsa44 protein-coding gene sequences were amplified, the purified amplicon was initially cloned into the pGEM-T Easy vector, and subsequently to the pETite vector for the expression of rLsa44 in Escherichia coli. After induction with 1 mM isopropyl-β-D-thiogalactoside (IPTG), the expressed recombinant protein with a molecular weight of 42 kDa was purified by the Ni-NTA affinity chromatography, characterized by SDS-PAGE and confirmed by Western blot using Leptospira specific sera. The sensitized latex beads (0.8 µm) coated with rLsa44 protein were prepared and assessed for their suitability as a diagnostic antigen in the LAT. Ten microliters of sensitized beads were used with test serum for agglutination, the results were recorded for 3 min and positive results were rated as +++, ++, +, if the clumps appeared after 1, 2, and 3 min, respectively. The rLsa44-LAT was evaluated for detection of anti-leptospiral antibodies using known Microscopic Agglutination Test (MAT) with positive (n = 74) and negative (n = 91) samples from apparently healthy animals, which revealed the relative diagnostic sensitivity (DSn) of 88

The present study was conducted to assess the microbial quality of water in forest waterholes in different seasons and its possible impact on wild animals, at Bandipur and Nagarahole Tiger Reserve forests in the state of Karnataka, India, during the year 2012 which evidenced drought, and the year 2014 which witnessed normal rainfall in these forests. The forests recorded the death of 39 wild elephants during April and May of 2012. One ailing elephant was confirmed to have high fever, diarrhoea, leucocytosis, and symptoms of colic. Water samples collected from major waterholes during the peak drought showed higher numbers of coliforms and several species of opportunistic bacteria including species of Vibrio and Campylobacter. In the year 2014–15, with normal rainfall, the death of less than 10 wild elephants was documented during April to May, 2015. We collected water samples from 20 major waterholes every month from June 2014 to May 2015 and assessed the water quality. We found that the microbial water quality improved in rainy season (June–September), started deterioration in winter (October–January) and became poor in summer (February–May). Though, the water during the summer of 2014–15 was equally of poor microbial quality as seen during peaks of droughts, the elephant deaths were relatively lower, signifying the role of normal rainfall in forests which provides the availability of fodder and water, which determines the general body condition and ability to resist opportunistic infections. We discuss the measures suggested and implemented from this study and their utilities at ground level.

The experiment was undertaken to evaluate the effect of pomegranate (Punica granatum) juice and peel extract on antioxidant status in streptozotocin induced diabetic rats. The study included seven treatment groups comprising of ten albino Wistar rats each. The various groups in this study included normal control Group I (NC), diabetic control (Group II (DC), diabetic rats treated with metformin at the rate of 500 mg/kg bw Group III (MF), diabetic rats treated with pomegranate fresh juice at 1mL/day Group IV(PJ), diabetic rats treated with pomegranate peel extracts at 100mg/day Group V (PPE), diabetic rats treated with pomegranate fresh juice 1mL + pomegranate peel extracts (100mg) at 50 per-cent dosage and metformin (half dose) Group-VI (PJ+PPE+MF 50%), diabetic rats treated with pomegranate fresh juice (0.5mL) + pomegranate peel extracts extract (50mg) at 50 per-cent dosage and metformin (half dose) (Group-VII (PJ+PPE @ 50% +MF 50%). Antioxidant status of rats was evaluated by estimating product of oxidative injury (MDA) and endogenous antioxidant enzymes (CAT, SOD and GPx) in the liver on 3rd, 15th, 30th and 45th day. All the antioxidant enzymes (CAT, SOD and GPx) markedly reduced in diabetic control (DC) rats. However, treatment groups treated with pomegranate juice or pomegranate peel extract (Group IV to VII) individually or in combination showed a significant improvement in all the antioxidant enzymes (CAT, SOD and GPx) compared to the diabetic control rats. It was observed that metformin substantially alleviated the effects of STZ in diabetic rats compared to all the treatment groups. Pomegranate peel extract was observed to have marginally improve antidiabetic effects compared to pomegranate juice. Combination of pomegranate juice and pomegranate peel extract with metformin at half dose and full dose also alleviated STZ induced diabetic effects significantly. However, there was no dose dependent and synergistic effect. It could be inferred that bioactive compounds of pomegranate juice and pomegranate peel extract have antiperoxidative effect due to their ability to scavenge free radicals or chelate metal ions in STZ induced rats.

Bovine mastitis is a persistent and major problem in the dairy industry globally. Staphylococci and Escherichia coli are predominantly responsible for this condition. The aim of this study was to look for the genes associated with antimicrobial resistance in these predominant bacteria isolated from bovine mastitis in different regions in Karnataka. In all, 211 bacterial isolates were recovered from bovine mastitis cases in Karnataka. Of these, 85, 23 and 47 were Staphylococcus aureus, S. epidermidis and E. coli, respectively, as confirmed by PCR. Among S. aureus, blaZ was detected in 61.17% (52/85), mecA and tetK detected in 3.52% (3/85), aacA-aphD, ermC and strA genes detected in 4.7% (4/85), tetM was detected in 16.4% (14/85) and strB in 7.05% (6/85) isolates. None of the isolates contained tetO and ermA genes and 30.58% (26/85) isolates did not reveal any of the genes. Among S. epidermidis, blaZ was detected 43.47% (10/23), mecA and tetM detected in 30.43% (7/23), aacA-aphD and ermC detected in 13.04% (3/23), tetK detected in 8.69% (2/23), strA detected in 4.34% (1/23) and strB detected in 21.73% (5/23) isolates. None of them had tetO and ermA genes and 13.04% (3/23) isolates did not reveal any of the genes screened. Among E. coli, blaTEM and tetA genes were detected in 100% (47/47), blaSHV and strA were detected in 31.9% (15/47), blaOXA 30 detected in 12.76% (6/47), tetM and tetB detected in 2.12% (1/47), and strB was detected in 51.06% (24/47). None of the isolates contained aacA-aphD, mecA, tetO, tetK, ermA and ermC genes. This genetic profile of prevalence of antimicrobial genes for varied antibiotics/antimicrobials indicates the possible wider antimicrobial resistance among the predominant species of bacteria causing bovine mastitis. Further, phenotypic characterisation of these isolates with respect to their antimicrobial resistance would be of significance.

BACKGROUND AND AIM:In recent times, non-aureus staphylococci (NAS) have emerged as the major organisms isolated from mastitis cases in dairy animals, with a predominance of Staphylococcus epidermidis and Staphylococcus chromogenes. As compared to Staphylococcus aureus, much less is known about the molecular types or the spatiotemporal epidemiology of these NAS species. In the present study, randomly amplified polymorphic DNA (RAPD) was employed to detect genetic polymorphisms, intraspecies diversity, and epidemiology of S. chromogenes strains (n=37) isolated from bovine and bubaline mastitis cases in the state of Karnataka.MATERIALS AND METHODS:Thirty-seven S. chromogenes isolates (14 from bovines and 23 from bubaline) isolated from subclinical mastitis cases, from organized and unorganized sectors, were subjected to RAPD typing. Further, methicillin resistance was determined by cefoxitin disk diffusion method.RESULTS:The amplified DNA fragments ranged from 150 to 3000 base pairs and yielded several RAPD profiles. Further analysis using Digital Image Correlation Engine correlation coefficient and UPGMA method showed that the 37 isolates could be classified into 12 distinct RAPD types (A to L) at 62% similarity (D=0.889). Four of the most predominant RAPD types, B, A, C, and E, in that order, and together, represented 65% of the isolates. High diversity was observed among the isolates both within farms and between geographic locations. Most of the isolates exhibited methicillin resistance. This is the first such report from India.CONCLUSION:In the absence of defined multilocus sequence type protocols or sufficient sequences available in the public domain, RAPD can be employed to determine genetic diversity of S. chromogenes isolates.

Bovine mastitis is of major economic importance and is treated by using various antimicrobials/antibiotics at the field level. However, antimicrobial resistance among the predominant mastitis causing bacterial pathogens is an issue in successful treatment. Also mastitis is increasingly becoming a public health concern due to the ability of the causative bacterial pathogens and/or their products, such as enterotoxins, to enter the food supply and cause food-borne diseases. The objective of this study was to assess the antimicrobial resistance (AMR) in Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli from bovine mastitis by employing microtiter plate based Minimal Inhibitory Concentration (MIC) of commonly used antimicrobials/antibiotics in Karnataka. In all, out of 211 bacterial isolates from bovine mastitis cases, S. aureus (n=85), S. epidermidis (n=23) and E. coli (n=47) isolates, which were confirmed by targeting species specific genes like nuc, rpoB and uspA, respectively were used. On in-vitro antimicrobial susceptibility testing of 31 S. aureus isolates for 6 different antimicrobials, 9.7 per cent (3/31) of isolates were found to be sensitive and 90.3 per cent (28/31) were resistant to penicillin, and 29 per cent (9/31) isolates were sensitive and 70.9 per cent (22/31) were resistant to cefotaxime. All of the 5, 9, 17 and 8 S. aureus isolates subjected to gentamicin, streptomycin, tetracycline and oxacillin MIC, respectively, were found resistant. Of the 10 S. epidermidis isolates used for susceptibility testing, 5 isolates each were subjected to penicillin, tetracycline and cefotaxime, 3 isolates each to gentamicin and streptomycin and 6 isolates for oxacillin susceptibility testing. All of them exhibited resistance to all the six antimicrobials. On MIC testing of 20 E. coli isolates, all of them showed resistance to penicillin, enrofloxacin and tetracycline; whereas 85 per cent (17/20) of isolates were sensitive and 15 per cent (3/20) were resistant to cefotaxime. Of 10 E. coli isolates subjected to streptomycin MIC, all of them showed resistance. Overall, the study revealed the high prevalence of AMR in S. aureus, S. epidermidis and E. coli to multiple antimicrobials used to treat bovine mastitis.

Antimicrobial resistance (AMR) in poultry production chain is one of the major food safety concerns due to indiscriminate usage of antibiotics and the presence of pathogens such as Salmonella which causes infections in various stages of production. In the present study, 182 samples were collected from commercial broiler supply chain, viz., three hatcheries (n=29), three commercial broiler farms (CBF; n=99), and three retail meat shops (RMS; n=54), and used for isolation and identification of Salmonella using three different selective agar media and a selective enrichment medium followed by PCR confirmation targeting the hilA gene. The overall prevalence of Salmonella was 47/182 (25.82%), and a significantly higher (P<0.05) prevalence was observed in retail meat shops (46.29%), CBF (19.19%), and hatcheries (10.34%). Comparison of three agar media for isolation of Salmonella revealed that all the media were equally selective. However, PCR amplification of hilA gene fragment was significantly higher (P<0.01) in selective enrichment culture tetrathionate brilliant green bile broth (TTB) as compared to all solid (agar-based) media. Susceptibility pattern against most frequently used antibiotics revealed that 100% of the isolates were resistant to at least one antibiotic. High resistance was observed for doxycycline (94.34%), followed by cefpodoxime (84.91%), ciprofloxacin (72.64%), gentamicin (65.09%), enrofloxacin (61.32%), colistin sulphate (40.42%), amikacin (34.91%), ampicillin (33.96%), neomycin (33.02), cefotaxime (30.19%), ceftazidime (29.25%), trimethoprim-sulfamethoxazole (23.58%), amoxicillin+clavulanic acid (21.70%), and chloramphenicol (12.26%); 16.98% of the isolates were ex-tended spectrum β-lactamase (ESBL) producers, and 76.41% were multidrug resistant (MDR). MDR Salmonella were significantly higher (P<0.01) in RMS (91.66%) followed by CBF (82.75%), whereas no MDR isolates were present in the isolates from hatcheries. The results indicated a higher prevalence of Salmonella and AMR for commonly used antibiotics in the complete broiler supply chain, especially RMS and CBF. Also, this study idicated that TTB enrichment followed by PCR and colony PCR was found to be rapid, specific and time-saving method.

Background: Brucellosis is an economically important zoonotic disease, affecting domestic animals and humans in many developing countries including India. Small ruminant brucellosis caused by B. melitensisis the primary source of human brucellosis. The B. melitensis Rev 1 vaccine, though considered being the best available vaccine against small ruminant brucellosis, it is not free from risk of handling. Hence, to address the biohazard nature of vaccine, the study aimed to evaluate the construction of B. melitensis ghost vaccine, an alternate to live vaccine. The ghost vaccine compared with standard B. melitensis Rev 1 vaccine along with challenge studies in mice model. Methods and materials: The bacterial ghosts were constructed using B. melitensis Rev 1 and B. melitensis field isolate at a cell concentration of 3.0 × 109 CFU/mL. The bacterial cells were lysed using GI24, a porcine cathelicidin at the rate of 80 μg/mL. The ghost formation was confirmed by absence of growth on brucella selective agar (BSA), non-amplification of 731 bp in AMOS PCR and morphological changes including tunnel formation under scanning and transmission electron microscopy. Two ghost vaccines and the standard Rev 1 vaccine were evaluated in mice by intra peritoneal route, for humoral immune response by quantifying IgG levels in post vaccinal sera and for cell mediated immune response by lymphocyte proliferation assay and profiling of Th2 (IL-4, IL-10) and Th1 (TNF-α, IFN-γ) cytokines. Results: The results revealed a significant (P < 0.05) difference in IgG levels between control and vaccinated groups. The interplay of Th2 and Th1 cytokine profile was characterised by an initial domination of Th2 response by elaborating IL-4 and IL-10 needed to antagonize the TNF-α and IFN-γ cytokines which is critical to initiate cell mediated immune response. In later stages there was decreased IL-4 and IL-10 levels, essential for TNF-α and IFN-γ surge for an effective Th1 mediated cellular immune response required for clearance of Brucella organisms. Vaccinated mice withstood the challenge by B. melitensis 16M. Conclusion: It is concluded that, although B. melitensis Rev 1 has an edge over ghost vaccines in eliciting immune response; the ghost vaccines appeared to be safe and offer an alternate putative vaccine candidates against brucellosis.

Dermatophytosis is a fungal skin infection which can infect various species of animals and human. The infection is also called ringworm and is caused by many types of fungi. In the present study total 633 small ruminants (441 from Noorgal district (188 sheep and 253 goats) and 192 from Goshta district (57 sheep and 135 goats)) and 320 humans in medical hospital were examined for presence of ringworm. Skin lesions of suspected patients were taken and subjected to KOH wet mount and culturing in Sabouraud's dextrose agar (SDA). Overall prevalence in small ruminants according to KOH test was 5.213 per cent and according to SDA culture was 5.529 per cent. In sheep the prevalence as per KOH and SDA culture was 6.93 per cent each. In goat KOH test showed 4.123 per cent and fungal culture showed 4.639 per cent positive. Result of medical hospital revealed that the overall result based on KOH was 4.68 per cent and on culture base was 3.75 per cent. In Goshta and Noorgal districts females were more susceptible to ringworm than the males and old sheep were more susceptible than young. In Goshta young goats were more susceptible whereas in Noorgal old goats were more susceptible. The findings revealed that there was high prevalence of dermatophyte both in human beings and animals in this part of Afghanistan.

6-Thioguanine encapsulated chitosan nanoparticles (6-TG-CNPs) has formulated by the ionic-gelation method. Morphologically, the 6-TG-CNPs were spherical and showed mean size, PDI, zeta potential, and entrapment efficiency of 261.63 ± 6.01 nm, 0.34 ± 0.10, +15.97 ± 0.46 mV and 44.27%, respectively. The IR spectra confirmed the 6-TG complex with chitosan. The in vitro drug release profile of 6-TG-CNPs revealed an increase in sustained-release (91.40 ± 1.08% at 48 h) at pH 4.8 compared to less sustained-release (73.96 ± 1.12% at 48 h) at pH 7.4. The MTT assay was conducted on MCF-7 and PA-1 cell lines at 48 h incubation to determine % cell viability. The IC50 values of 6-TG, 6-TG-CNPs, and curcumin for MCF-7 were 23.09, 17.82, and 15.73 μM, respectively. Likewise, IC50 values of 6-TG, 6-TG-CNPs, and curcumin for PA-1 were 5.81, 3.92, and 12.89 μM, respectively. A combination of 6-TG-CNPs (IC25) with curcumin (IC25) on PA-1 and MCF-7 showed % cell viability of 43.67 ± 0.02 and 49.77 ± 0.05, respectively. The in vitro cytotoxicity potential in terms of % cell viability, early apoptosis, G2/M phase arrest, and DNA demethylating activity of 6-TG-CNPs alone and combination with curcumin proved to be more effective than that of 6-TG on PA-1 cells.