A multipronged approach for the detection of leptospirosis in captive sloth bears (Melursus ursinus) in Agra and Bannerghatta sloth bear rescue centers in India

Leptospirosis is an exacerbating factor responsible for the drastic decline of sloth bear population in India. In this study, a multipronged approach based on antigen detection using Polymerase Chain Reaction (PCR) employing G1/G2 and LigBF/LigBR primers, antibody detection using Microscopic Agglutination Test (MAT) and recombinant LigBCon1-5 antigen based Latex Agglutination Test (rLigBCon1-5 LAT), serum biochemistry using hepatic (serum glutamate oxalo acetic transaminase (SGOT) and serum glutamate pyruvic transaminase (SGPT) and renal biomarkers (blood urea nitrogen (BUN) and Creatinine) and gross/histopathological evidence in liver and kidneys were employed to investigate leptospirosis in captive sloth bears. A total of 133 serum samples collected from Agra (n=113) and Bannerghatta (n=20) sloth bear rescue centers were screened using MAT and rLigBCon1-5 LAT. A total of 87 and 78 sera tested positive by MAT and LAT respectively. Pyrogenes was the leading serovar obtained using MAT followed by Icterohaemorrhagiae, Javanica, Grippotyphosa, Canicola and Tarassovi. The relative sensitivity, specificity and accuracy of rLigBCon1-5 LAT in comparison to MAT were 89.66%, 100% and 93.23% respectively. PCR performed on hepatic and renal tissues showed amplicon of 285 and 219 base pairs for G1/G2 and LigBF/LigBR primers respectively. Gross evidence (icteric liver, severely engorged hepatic sinusoids, congested kidneys with necrotic white spots on sub capsular surface), histopathology (severe hepatic degeneration and tubulointerstitial nephritis) and elevated hepatic/renal biomarkers were suggestive of leptospirosis. This study suggests that rLigBCon1-5 LAT can be employed as a pen-side test for detecting leptospirosis in sloth bears.     

Emergence of antigenic variants with in serotype A foot and mouth disease virus in India and evaluation of a new vaccine candidate panel

Emergence of genetically and antigenically divergent lineages/genotypes and poor intergenotypic antigenic coverage is a major concern in serotype A foot-and-mouth-disease virus (FMDV) in India. In 2009, to cover antigenic diversity emerged in serotype A virus field isolates, IND40/2000 was selected as the new vaccine strain for incorporation in the trivalent FMD vaccine formulation used in India. Although current vaccine strain (IND40/2000) covers most isolates antigenically, a few VP3(59)-deletion group isolates showed low r-value in routine vaccine matching exercise. The VP3(59)-deletion group within genotype 18 emerged first in late part of 2002 and in 2007 causing outbreaks along with non-deletion isolates of the same genotype. In case of emergence or re-emergence of more antigenically divergent isolates in future, a need for a new vaccine candidate to cover maximum isolates of both deletion and non-deletion group may arise. Four alternate candidate vaccine strains (IND281/2003, IND195/2007, IND360/2007 and IND123/2008) were selected based on set criteria and antigenic relationships with field isolates sampled between 2002 and 2009 were analyzed using a micro-neutralization test. Phylogenetic analysis based on capsid region of serotype A isolates revealed existence of two broad distinct clusters (VP3(59)-deletion and non-deletion group) within genotype 18. The VP3(59)-deletion group has diversified genetically with time giving rise to three different sub-lineages (clade18a, 18b and 18c). The present study indicates that the virus candidates IND281/2003 (VP3(59)-deletion group) and IND195/2007 (non-deletion group) can be used as an adjunct or alternative strain to currently used vaccine strain IND40/2000 in case of emergence of more antigenically divergent isolates in future.     

Natural compounds enhancing growth and survival of rhizobial inoculants in vermicompost-based formulations

The present study demonstrates the usefulness of natural microbial growth-promoting compounds for improving the stability and life of vermicompost-based (both granular and its aqueous extract) bioformulations. Granular vermicompost maintained the number of cells of Rhizobium meliloti Rmd 201 up to 5.9 × 10⁸ after 180 days at 28°C compared with 2.1 × 10⁸ in charcoal (powdered), while aqueous extract of the vermicompost supported the 5.6 × 10⁷ rhizobia numbers even after 270 days. The addition of 25 μL/mL cow urine and 0.01 mM calliterpinone, a natural plant growth promoter, increased the rhizobia number significantly in granular vermicompost and its aqueous extract, respectively.     

Evaluation of well-being, productivity, and longevity of pregnant sows housed in groups in pens with an electronic sow feeder or separately in gestation stalls

OBJECTIVE: To compare well-being, performance, and longevity of gestating sows housed in stalls or in pens with an electronic sow feeder (ESF). ANIMALS: 382 pregnant sows of parities 1 through 6. PROCEDURE: Sows were housed in separate stalls (n = 176) or group pens (206) with an ESF. Well-being of sows was assessed at various time points in terms of injuries, salivary cortisol concentration, and behavior in a novel arena or to a novel object. Farrowing performance and longevity of sows were also assessed. RESULTS: Total injury scores (TIS) of sows in pens were significantly higher at initial introduction and mixing. In stall-housed sows, TIS was significantly higher during late gestation. The TIS and cortisol concentration were significantly lower in stall-housed sows, compared with values for sows in pens. As parity increased, the likelihood of higher median TIS decreased significantly in pen-housed sows and increased significantly in stall-housed sows. The TIS of sows in pens was negatively correlated with body weight and backfat thickness, whereas these correlations were positive in stall-housed sows. Farrowing performance and results for novel arena or objects did not differ. Proportion of sows removed was significantly higher for pens than for stalls; lameness was the major reason for removal for both systems. CONCLUSIONS AND CLINICAL RELEVANCE: Stalls impose space restrictions for larger sows, resulting in injuries during late gestation. Interventions are needed to minimize aggression during initial introduction and mixing and at the ESF in pens to reduce severe injuries or lameness of gestating sows.     

Comparison of injuries in sows housed in gestation stalls versus group pens with electronic sow feeders

OBJECTIVE: To compare the location and severity of injuries in pregnant sows housed in individual gestation stalls with that in pregnant sows housed in dynamic groups in pens with electronic sow feeders. DESIGN: Prospective study. ANIMALS: 100 pregnant sows of parity 1 to 3 and various body weights. PROCEDURE: Fifty sows each were randomly allotted to gestation stalls or group pens with electronic sow feeders. Injuries were scored on the basis of location, number, and depth of wounds. Injury scores for sows in both housing systems were compared during a period of 90 days. The influence of factors such as duration of stay in the housing system, parity, and body weight on sow injuries was also examined. RESULTS: Injury scores were higher in group pens with electronic sow feeders. As body weight increased, injury scores decreased for sows housed in group pens with electronic sow feeders and increased for sows housed in gestation stalls. There was a significant negative association between second parity and total injury scores. CONCLUSIONS AND CLINICAL RELEVANCE: Modifications in housing system design and management procedures are needed to reduce injuries in pregnant sows.     

Attenuated cytokine responses in porcine lymph node cells stimulated with CpG DNA are associated with low frequency of IFNalpha-producing cells and TLR9 mRNA expression

The immune stimulatory effects of synthetic CpG DNA, on porcine peripheral blood mononuclear cells (PBMC) have been reported, but little is known about CpG-induced responses in other lymphoid tissues of pigs. We investigated innate immune responses induced by CpG DNA in cells from blood, lymph nodes (LN) and spleens of pigs. Porcine PBMC and lymph node cells (LNC) were stimulated in vitro with three classes (A-, B- and C-class) of CpG oligodeoxynucleotides (ODNs), and a non-CpG control ODN. All three classes of CpG ODNs induced significant production of IFNalpha, TNFalpha, IL-1, IL-6 and IL-12 in PBMC. In contrast, in LNC, only IL-12 was stimulated by all three classes of CpG ODNs, while IFNalpha, and IL-6 were induced by A- and C-class ODNs. No TNFalpha was induced in LNC by any of the ODNs. Significant lymphocyte proliferation was induced in PBMC by all three classes of CpG ODNs and non-CpG control. However, in LNC, B- and C-class ODNs induced significant proliferation, while no proliferation was seen with A-class and non-CpG control ODN. All three classes of ODNs induced NK-like cytotoxicity in PBMC and spleen cells, but were less effective in inducing NK cytotoxicity in LNC. We then investigated the reasons for the relatively poor CpG-induced responses in LNC. Our investigations revealed that LNC had a lower frequency of IFNalpha-secreting cells and expressed low levels of TLR9 mRNA compared to PBMC. We conclude that the lower number of IFNalpha-secreting cells and receptor expression may contribute to the attenuated responses in LNC following stimulation with CpG ODN.     

Rice Bran Oil: Emerging Trends in Extraction,Health Benefit,and Its Industrial Application

Rice bran oil(RBO) is unique among edible vegetable oils because of its unique fatty acid composition, phenolic compound(γ-oryzanol, ferulic acid) and vitamin E(tocopherol and tocotrienol). It has become a great choice of cooking oil because of its very high burning point, neutral taste and delicate flavour. Non-conventional methods of RBO extraction are more efficient and environmentally friendly than conventional extraction methods. Advances in RBO extraction using innovative extraction strategies like super/sub-critical CO_2, microwave-assisted, subcritical H_2 O, enzyme-assisted aqueous and ultrasoundassisted aqueous extraction methods have proven to significantly improve the yields along with improved nutritional profile of RBO. The compositions and strategies for stabilization of RBO are well discussed. The constituents are present in the RBO contribute to antioxidative, anti-inflammatory, antimicrobial, antidiabetic and anti-cancerous properties to RBO. This has helped RBO to become an important substrate for the application in food(cooking oil, milk product and meat product) and non-food industries(polymer, lubricant, biofuel, structural lipid and cosmetic). This review provided comprehensive information on RBO extraction methods, oil stabilization, existing applications and health benefits.     

Efficacy of single and multiple doses of fenbendazole against gill parasites (Dactylogyrus sp.) of Labeo rohita (Hamilton, 1822) and its physio‐metabolic effects on the fish

The study was conducted to investigate the efficacy and physio‐metabolic responses of single and multiple doses of fenbendazole (FBZ) in Labeo rohita against gill parasite (Dacylogyrus sp.). Two Isoenergetic (395 Kcal DE/100 g) and isonitrogenous (35% CP) purified diets viz. control (without FBZ) and treatment (0.2% FBZ) were prepared. The efficacy of FBZ was measured after administration of single dose (20 mg/kg b. wt.) on 0th day and multiple doses of FBZ, that is, 20 mg/kg b. wt. on 0th, 3rd and 7th day through diet. Although oral administration of FBZ at single dose could significantly reduce the parasite in fish in comparison with the control group, 100% efficacy of the drug was not achieved up to 15th day of treatment. On the other hand, multiple doses of FBZ could show almost 100% efficacy on 15th day in terms of elimination of parasites from gill. The drug induced oxidative stress in examined fish. There was no significant change in activities of hepatic and muscle aspartate aminotransferase and alanine aminotransferase of L. rohita up to 15th day after oral administration of FBZ at single dose. But both single and multiple doses of FBZ could significantly enhance the hepatic superoxide dismutase, catalase and glutathione‐S‐transferase activities in fish up to 7th day of drug administration and significantly decreased thereafter up to 15th and/or 30th day. It is concluded that oral administration of FBZ at multiple doses (20 mg/kg b. wt.) on 1st, 3rd and 7th day could be effective against Dactylogyrus sp. infection in L. rohita.