Bablu Kumar

Predominance of rotavirus genotype G6P[11] in diarrhoeic lambs

Out of 500 faecal samples from lambs with diarrhoea in Jammu and Kashmir, India, 66 (13.2%) were positive for group A rotavirus (GARV) by the latex agglutination test (LAT). Electropherotyping by RNA-polyacrylamide gel electrophoresis revealed the typical GARV 4-2-3-2 migration pattern in 49/66 (74.2%) samples. Fifty-two samples (10.4%) were positive by reverse transcription-PCR. G6 was the predominant G genotype (25/52; 48.07%), followed by G10 (19/52; 36.54%) whereas, the predominant P genotype was P[11] (46/52; 88.46%). G6P[11] is the prevalent strain of group A rotavirus in sheep in Jammu and Kashmir, India.

Bacteriological and Molecular Detection of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in Equines of Northern India

Present study was undertaken to study the prevalence of β-haemolytic streptococci in equine of northern temperate region of Jammu and Kashmir, India. One hundred and forty one samples were collected in duplicate from nasopharyngeal tract of diseased (53) and apparently healthy equine (88) for isolation and direct PCR. A total of 77 isolates of streptococci were recovered from 141 samples with an overall prevalence rate of 54.60%. Out of these 77 isolates, 52 were from diseased and 25 from apparently healthy animals. Of the 77 isolates, 4 were identified as Streptococcus equi subsp. equi, 56 as S. equi subsp. zooepidemicus and 17 as S. dysgalactiae subsp. equisimilis. Thus the overall prevalence of S. equi subsp. equi, S. equi subsp. zooepidemicus and S. dysgalactiae subsp. equisimilis was 2.83, 39.71 and 12.05% respectively. The sensitivity of the PCR for the detection of S. equi species was found higher when attempted from direct swab samples.

Prevalence and Antibiogram study of Rhodococcus equi in equines of Jammu and Kashmir, India.

ABSTRACT The present study was conducted to determine the prevalence of Rhodococcus equi infection in equines of Jammu and Kashmir, India, and evaluate the zoonotic threat posed by this organism to equine owners and tourists. One hundred and forty-one samples (98 samples from adult animals ≥5 years old and 43 samples from foals less than 6 months old) were collected in duplicate from nasopharyngeal tract of equines for isolation and direct PCR. A total of 12 isolates of R. equi were recovered, of which 9 were from foals and 3 from adult animals. Therefore, the present study recorded prevalence rates of 20.93% and 3.06% among foals and adult equines respectively. The prevalence rates were found to be 25.58% and 4.08% by 16S rRNA species-specific PCR among foals and adult animals respectively. Thus, the PCR-based assay was found to be more sensitive and helped in quick detection of R. equi than the culture based method which is time consuming and laborious. However, the culture-based method is still preferred due to some limitations of PCR. The antibiogram of the isolates revealed that erythromycin and rifampicin were the most effective antimicrobials with 100% sensitivity, followed by amoxicillin (66.67%), lincomycin (58.3%) and kanamycin (58.3%). The results also revealed that resistance was highest for penicillin G (50%), followed by kanamycin (25%) and streptomycin (25%).

Protective immune-response of aluminium hydroxide gel adjuvanted phage lysate of Brucella abortus S19 in mice against direct virulent challenge with B. abortus 544.

The prophylactic efficacies of plain and alum adsorbed lysate were evaluated by direct virulent challenge in mice model. A recently isolated brucellaphage ϕLd was used for generation of lysates. Twenty four h incubated Brucella abortus S19 broth cultures standardized to contain approximately 10(8)xa0CFU/ml were found suitable for generation of lysates. Three lysate batches produced through separate cycles did not show any significant variation with respect to protein and polysaccharide contents, endotoxin level and phage counts, indicating that compositionally stable lysate preparations can be generated through an optimized production process. Three polypeptides of ∼16, 19 and 23xa0kDa could be identified as immuno-dominant antigens of the lysate which induced both humoral and cell-mediated immune responses in a dose dependent manner. Results of efficacy evaluation trial confirmed dose-dependent protective potencies of lysate preparation. The lysate with an antigenic dose of 0.52xa0μg protein and 60xa0μg CHO adsorbed on aluminium gel (0.1 percent aluminium concentration) exhibited the highest protective potency which was greater than that induced by standard S19 vaccine. Phage lysate methodology provides a very viable option through which an improved immunizing preparation with all desirable traits can be developed against brucellosis, and integrated with immunization programmes in a more efficient manner.

Active immunization with Brucella abortus S19 phage lysate elicits serum IgG that protects guinea pigs against virulent B. abortus and protects mice by passive immunization

Brucellosis is an economically important zoonosis of worldwide significance. Earlier (Jain etxa0al., 2015) we reported methodology for generation of phage lysate preparations against Brucella abortus S19 using brucellaphage ϕLd. In this study, using a fixed dose (Two mouse PD100) of lysates, the prophylactic efficacies of both plain and alum gel adjuvanted lysates were evaluated in guinea pig by direct virulent challenge and passive mouse protection test (PMPT). Strong humoral and cell mediated immune responses in guinea pigs and protection comparable to S19 vaccine was observed with low dose (1.0xa0μg protein and 120xa0μg carbohydrate adsorbed on 0.1% aluminium gel). Passive transfer of antibodies to mice using d 90 post immunization sera of guinea pig protected the animals against challenge. The study suggested the significance of humoral immunity in murine brucellosis. Further, the methodology can be explored to produce a new class of immunotherapeutic agents against bovine brucellosis.

Draft genome sequence of field isolate Brucella melitensis strain 2007BM/1 from India

OBJECTIVESnBrucellosis is among one of the most widespread important global zoonotic diseases that is endemic in many parts of India. Brucella melitensis is supposed to be the most pathogenic species for humans. Here we report the draft genome sequence of B. melitensis strain 2007BM/1 isolated from a human in India.nnnMETHODSnGenomic DNA was extracted from Brucella culture and was sequenced using an Illumina MiSeq platform. The generated reads were assembled using three de novo assemblers and the draft genome was annotated.nnnRESULTSnThis monoisolate, with a genome length of 3268756bp, was found to be resistant to azithromycin and trimethoprim/sulfamethoxazole but susceptible to tetracycline, ofloxacin, rifampicin, ciprofloxacin and doxycycline. The presence of virulence genes in the strain was identified.nnnCONCLUSIONSnThe results obtained will help in understanding drug resistance mechanisms and virulence factors in highly zoonotic B. melitensis and suggest the need for judicious use of antibiotics in livestock health and management practices.

Rapid visual isothermal nucleic acid-based detection assay of Brucella species by polymerase spiral reaction

The aim of this study was to develop polymerase spiral reaction (PSR) for rapid, sensitive and specific detection of Brucella sp.

Immunization with Salmonella Abortusequi phage lysate protects guinea pig against the virulent challenge of SAE-742

Salmonella Abortusequi causes important clinical diseases in horses possibly leading to abortion. In the present investigation, the protective efficacy of both plain and aluminum hydroxide gel adjuvanted phage lysate was evaluated in guinea pig model. Broad host range bacteriophage PIZ-SAE-2, was characterized and used for generation of lysates. Three different lysate batches, produced through separate cycles and characterized, were pooled together for immunization study. Plain and adjuvanted phage lysate preparations elicited both humoral and cellmediated immunity. The adjuvanted lysate at a dose of 50u202fμl elicited the highest protective efficacy against direct challenge at 28th DPI. Thus, the present study describes a new method of bacterial inactivation for producing a new class of better & safe immunprophylactic agents. This is the first report of producing an inactivated vaccine candidate using a new approach against equine salmonellosis.

Protective Efficiency of Pasteurella multocida A:1 Bacteriophage lysate Vaccine against Homologous and Heterologous Challenge in Poultry

Fowl cholera (FC) caused by serotypes of Pasteurella multocida includes A:1, A:3, A:4 is a highly fatal septicemic disease. Preliminary trials of P. multocida A:1 bacteriophage lysate vaccine against FC was evaluated. Lytic phage and P. multocida ratio was standardized to obtain stable lysate batches. Consequently, three batches of lytic phage preparation were produced; estimation of protein and carbohydrate content amongst batches did not shown any significant variation indicating same batches can be produced by standardized procedure. Protective response trials in poultry with P. multocida A:1, A:3, A:4 against plain lysate and alum adsorbed lysate (1% alum) on vaccinated group showed both homologous and heterologous protection compared with inactivated whole cell group provided only homologous protection. Assessment of antibody response towards P. multocida A:1, A:3, A:4 antigen evaluated by Indirect Haemagglutination test (IHA) reveals presence of protective antibody titer.

Effect on immune response against Pasteurella multocida capsular type a:1 and a:4 on supplementation with protein purified derivatives of Mycobacterium bovis in chicken

The adjuvant potential of tuberculo-protein of Mycobacterium bovis and Montanide ISA-206 adjuvants in different combination with Pasteurella multocida serotype A:1 and A:4 whole cell antigens have been evaluated in chicken model. Four different groups of chicken were immunized with different combination of adjuvants and humoral immune response was assessed by indirect ELISA. The immune response study revealed that chicken immunized with whole cell antigen in combination with Protein precipitated derivative (PPD) as well as montanide ISA-206 elicited a robust humoral immune response. To assess the protective ability two groups of chicken immunized with P.multocida A:1 were challenged with 1 MLD of Pasteurella multocida A:1; whereas two other groups of chicken immunized with P.multocida A:4 bacterin were challenged with 100 MLD of P.multocida A:4. Challenge studies indicated that both the groups of chicken immunized with P. multocida A:1 whole cell antigen adjuvanted with both Montanide ISA-206 and Mycobacterium bovis PPD as well as only Montanide ISA-206 conferred 100 % protection against P multocida A:1. However two other groups immunized with P.multocida A:4 bacterins could not sustain 100 MLD dose of direct challenge and there were 100% death of all the birds in both the immunized group, but the group immunized with both Montanide and PPD showed delayed death. * Corresponding author