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Brucellosis is a widely prevalent zoonotic disease of major public health significance. A collection of Brucella melitensis and Brucella abortus field isolates of animal and human origin were subjected to MLVA-15 typing followed by phylogeography studies. The MLVA-15 analysis of B. melitensis (n = 65) field isolates resulted in 48 different profiles. The panel I marker bruce45 was found to be most conserved, while the rest of the panel I markers showed low to moderate length polymorphism. Among the panel II markers, bruce04, bruce16 and bruce30 showed a high discriminatory index. The MLVA-15 typing of 13 B. abortus field isolates revealed 13 different genotypes with panel II markers showing higher discriminatory ability vis-a-vis panel I. The minimum spanning tree analysis (MST) in comparison with isolates from the international database revealed that all B. melitensis and B. abortus isolates from this study belonged to the 'Eastern Mediterranean' and the 'abortus C' lineage, respectively. The MLVA-15 typing could differentiate field isolates of B. abortus and B. melitensis originating from different regions, reaffirming the technique's potential of high resolution and suitability for local epidemiological studies. The MLVA scheme also has the advantage of comparison of local isolates with a worldwide database, allowing for phylogeographical studies.

Campylobacteriosis is among the most frequently reported foodborne zoonoses. A total of 848 samples were screened for Campylobacter spp. and occurrence was found to be 8.7%, 2.3% and 1.65% in broiler cecum samples, chicken meat samples and slaughter house environment swabs, respectively. High level of antimicrobial resistance was found against tetracycline (64.1%), doxycycline (54.4%), ampicillin (46.6%), nalidixic acid (42.7%), kanamycin (35.9%), and ciprofloxacin (33.33%). Resistance to co-amoxiclav (19.4%) and erythromycin (21.4%) was less common. The MAR index of the isolates was in the range of 0.11-0.78. Multi-drug resistance was observed in 54.4% of the isolates, with 53.2% C. jejuni and 55.3% C. coli isolates found resistant against three or more classes of antimicrobials. Presence of mutations in gyrA and 23S rRNA genes was investigated, which revealed that all the fluoroquinolone resistant isolates possessed Thr-86-Ile point mutation, whereas only 68% of erythromycin resistant isolates had A2075G mutation. The tetO gene was present in 91.7% tetracycline resistant isolates and blaOXA-61 gene was detected in 97.9% of the ampicillin resistant isolates. The occurrence of antimicrobial resistant Campylobacter spp. in broiler chicken samples and slaughter house settings is a public health risk and calls for judicial use of antimicrobials.

Swine is considered as a suitable sentinel to predict Japanese encephalitis virus (JEV) outbreaks in humans. The present study was undertaken to determine the circulating genotypes of JEV in swine population of India. A total of 702 swine serum samples from four states of western, northern, northern-temperate, and north-eastern zones of India were screened by real-time RT-PCR targeting envelope gene of JEV, which showed positivity of 35.33%. The viral copy number ranged from 3 copies to 6.3 × 10

Japanese encephalitis (JE) is a mosquito borne flaviviral zoonoses, causing fatal disease in equines and humans. JE is endemic in most of the states of India with occurrence of human cases every year. The horses are not vaccinated against JE in India and thus they are at more risk of acquiring the disease. Due to non-availability of indigenously developed ELISA and high cost of imported kits, regular sero-surveillance is not being carried out to assess the true picture of JE virus in equine population of India. Therefore, a recombinant NS1 protein based indirect IgG ELISA was developed with the objective to assess the sero-positivity of JE virus in equine population of India. The diagnostic sensitivity and specificity of developed ELISA was 84.73 % and 86.70 %, respectively. The validation studies revealed good reproducibility of ELISA with kappa value ranging from 0.75 to 1 between the results of different laboratories. A total of 2069 horse serum samples were screened using the developed ELISA and 401 samples were positive for IgG against JEV with an overall sero-positivity of 19.38% in equine population of India. A sero-positivity of 25.90% and 12.22% was recorded in Himachal Pradesh and Jammu-Kashmir, both hill states of North zone of India for the first time, revealing the spread of virus to the non-endemic parts of the country. The high sero-positivity of JE virus recorded in equine population warrants the need for initiation of vaccination of horses in India to prevent the morbidity and mortality.

Brucellosis is a widely prevalent zoonotic disease of major public health significance. A collection of

Japanese encephalitis (JE) is a major public health problem in the South Asian countries including India. Pigs serve as a relevant sentinel model, the surveillance of which could predict a potential JE outbreak in human population nearby. However, existing serological detection methods like Enzyme-Linked Immuno Sorbent Assay (ELISA), virus neutralization test (VNT) and Haemagglutination Inhibition (HI) require elaborative laboratory facilities which are invariably not available in field conditions. Recognizing the lacunae, attempts were made to develop recombinant antigen (rNS1) based latex agglutination test (LAT) as a rapid on-site test using covalent coupling method. Four different formats were evaluated using different coupling buffers, blocking buffers and reaction conditions. The format in which borate buffer at alkaline pH (8.5) was used for coupling of antigen with carboxylated beads followed by blocking with skimmed milk powder was found to be the best amongst all. Developed latex based test was used for screening of 207 pig serum samples for JE which revealed relative diagnostic sensitivity and specificity of 80.2% and 95.2%, respectively in comparison with indirect IgG ELISA. Hence, the present study demonstrated that covalently coupled recombinant antigen based LAT could be used as a reliable screening test for surveillance of JE in pigs under field conditions.

Purpose This paper aims to explore the application of drumstick (Moringa oleifera) flower (DF) as a functional antioxidative ingredient in goat meat product. Design/methodology/approach Dried DF was included in the product formulation at 1% (Treatment I) and 2% (Treatment II) levels. The physicochemical, colour, textural and sensory quality as well as storage stability of nuggets with DF were determined against control. Findings The dried DF was found to be rich source of protein and dietary fibre, possessing good antioxidant potential. Chromatographic analysis of DF extract showed presence of 14 active principles known to have antioxidative properties. Inclusion of dried DF decreased pH values of emulsion (p = 0.005) as well as nuggets (p p p = 0.017), yellowness (p p p = 0.032), cohesiveness (p = 0.006), gumminess and chewiness (p p = 0.002) flavour score for Treatment II. DF inclusion lowered (p Research limitations/implications DF can be used as a source of antioxidants and dietary fibre in goat meat nuggets to enhance their health value, functionality and storage stability. Originality/value Foods including goat meat nuggets enriched with goodness of functional ingredients like dietary fibre and natural antioxidants are gaining consumer’s preference globally. Inclusion of drumstick flower in goat meat nuggets significantly increases the dietary fibre and antioxidants making such products healthier and more stable. Consumption of goat meat nuggets added with drumstick flower is expected to improve consumer’s well-being as well.

Japanese encephalitis (JE) is a re-emerging mosquito-borne zoonotic flaviviral disease. Swine sero-convert 2-3 weeks before infection occurs in humans and thus serves as a suitable sentinel for JE surveillance and outbreak prediction in human population. The present study was conducted with the objective of developing a lateral flow assay (LFA) for detecting JEV antibodies in swine sera. Three different formats were tried using recombinant NS1 protein as antigen in order to select the best format. In format I, gold nanoparticles were conjugated with antigen followed by spotting of antigen on NCM as test line and anti-antigen IgG on NCM as control line. In format II, gold nanoparticles were conjugated with antigen followed by spotting of staphylococcal protein A as test line and anti-antigen IgG as control line. Format III used gold nanoparticles conjugated with goat anti-pig IgG followed by spotting of antigen as test line and pig IgG as control line. Amongst the three formats, format II was found to be superior with 100% relative diagnostic sensitivity and 100% relative diagnostic specificity during monsoon and post-monsoon period. A panel of 500 field swine serum samples was tested using format II which revealed sero-positivity of 15.6%, and the format was found suitable to screen swine serum samples during monsoon and post-monsoon period.

Campylobacteriosis is among the most frequently reported foodborne zoonoses. A total of 848 samples were screened for

Japanese encephalitis (JE) is a mosquito-borne flaviviral zoonotic disease and is one of the major causes of encephalitis in children. Swine, being an amplifier host of Japanese encephalitis virus (JEV), play an important role in its epidemiology. Therefore, early detection of either JEV or antibodies against JEV in swine is a feasible alternative for initiating necessary measures to prevent the spread of infection to humans. Since IgM antibodies appear early in swine sera, recombinant NS1 protein based indirect IgM ELISA was developed in the present study with the objective to know the recent infection of swine population with JEV. The relative diagnostic sensitivity and specificity of the developed ELISA was 95.34% and 98.6%, respectively. The developed ELISA was found to have excellent reproducibility on inter-laboratory and inter-institutional validation studies. A total of 3,027 field swine sera samples were screened using the developed ELISA and 488 samples were found positive for IgM against JEV with an overall sero-positivity of 16.12% in swine population of India. The highest sero-positivity was observed in swine population of Eastern zone of India which coincided with the maximum number of human JE cases reported from this zone during the same period. Further, antibody kinetics study revealed that the IgM antibodies against NS1 protein of JEV started appearing in swine sera at day 5 and disappeared completely by day 40. The IgG antibodies started appearing at day 7, and remained for more than 365 days indicating the suitability of IgM ELISA to know the recent infection of JEV. The developed IgM ELISA can be readily incorporated into surveillance programs for detection of JEV activity in swine population so that outbreaks in humans can be prevented by taking suitable preventive measures.

Japanese encephalitis (JE) is an important viral zoonotic disease in Asia, especially in rural and suburban areas where rice cultivation and pig farming coexist. Pigs serve as a suitable sentinel model, the surveillance of which could predict a potential JE outbreak in human population in the immediate vicinity. However, existing diagnostics like ELISA and VNT require sophisticated laboratory facilities which are more often not available in field conditions. In the present study, we aimed at developing recombinant non-structural (NS1) protein-based dipstick IgG ELISA as an on-site assay for sero-diagnosis of JE in swine. The assay was standardized by optimizing various parameters and the following conditions were found to be ideal including 1 μg of rNS1 protein in carbonate buffer per strip of nitrocellulose membrane comb; bovine serum albumin as blocking agent at 4 °C overnight; serum dilution of 1:10 and conjugate dilution of 1:5000 in skimmed milk powder. Relative diagnostic sensitivity and specificity of dipstick IgG ELISA was 100% and 92.9%, respectively. The dipstick assay was validated in three laboratories as per OIE guidelines. The storage life of dipstick was up to 7 months at 4 °C. The assay is easy to perform and the results can be interpreted with visual observation that precludes the need for absorbance reading equipment. The standardized dipstick assay was found promising for screening swine serum samples in field conditions. Timely detection of JE virus in swine will aid in predicting the outbreak in humans and thus in taking suitable preventive and control measures.

Japanese encephalitis is an emerging mosquito-borne flaviviral zoonotic disease. The present study was undertaken with the objective of developing rapid and sensitive nucleic-acid-based assays for detection of Japanese encephalitis virus (JEV) in swine blood samples. Three nucleic-acid-based assays, viz., reverse transcription polymerase chain reaction (RT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP), and real-time RT-PCR, were developed and compared in terms of their diagnostic efficacy. All three assays were found to be 100 per cent specific. The minimum detection limit of RT-LAMP and real-time RT-PCR was 12 copies/µl, while RT-PCR could detect 1.2 × 10(5) copies/µl. On comparison, RT-LAMP and real-time RT-PCR were 4-log more sensitive than RT-PCR. The applicability of the assays was evaluated by screening 135 field swine blood samples, of which 24 (17.77 %) were positive by RT-LAMP and real-time RT-PCR and only six (4.44 %) were positive by RT-PCR. The viral load in swine blood samples ranged between 2 × 10(6) and 4.8 × 10(9) copies per ml of blood by real-time RT-PCR. The comparative diagnostic sensitivity and specificity of RT-LAMP vis-à-vis real-time RT-PCR was found to be 100 %, while the sensitivity and specificity of RT-PCR vis-à-vis real-time RT-PCR was found to be 25 % and 100 %, respectively. Thus, the use of RT-PCR may cause the incidence of JEV in the swine population to be underestimated, while the real-time RT-PCR reported here is the test of choice for reference laboratories, and the newly developed one-step RT-LAMP assay will be suitable for field-level testing.

The present study was designed to determine the occurrence of C. jejuni and C. coli in chicken meat, chicken offal, chevon and milk samples from Bareilly, Uttar Pradesh (India) to elucidate the role of these products in transmission of Campylobacter to humans. The samples were examined using both cultural as well as polymerase chain reaction based detection method. Out of 280 samples examined, 29 (10.36%) samples were found positive by cultural method, where as 68 (24.29%) samples showed presence of Campylobacter species by polymerase chain reaction. All the samples depicted positive by culture were also found positive by PCR. The highest occurrence rate was observed in chicken meat (46%), followed by chicken offal (20%), chevon (18%) and milk (3.75%). The results depicted the superior efficacy of PCR for rapid screening of samples owing to its high rapidity, specificity and automation potential. The study suggested the potential of transmission of Campylobacter species to humans due to the consumption of raw or contaminated meat or milk.