Characterization of anthocyanins in grape juices by ion trap liquid chromatography-mass spectrometry

A reverse phase HPLC and electrospray interface with ion trap mass spectrometer method was developed for the characterization of anthocyanins in Concord, Rubired, and Salvador grape juices. Rubired and Salvador grapes are hybrids from Vitis vinifera and Vitis rupestris. Concord grape is a grape from the native American cultivar Vitis labrusca. Individual anthocyanins in these three varieties were identified on the basis of UV-vis and MS spectra and further elucidated by MS/MS spectra. Anthocyanins in Salvador and Concord grapes were 3-O-glucosides, 3-O-(6' '-O-p-coumaroyl)glucosides, 3-O-(6' '-O-p-acetyl)glucosides, 3,5-O-diglucosides, and 3-O-(6' '-O-p-coumaroyl)-5-O-diglucosides of delphinidin, cyanidin, petunidin, peonidin, and malvidin. Vitisin B was detected in Salvador grape juice. Anthocyanins in Rubired grape juice were primarily anthocyanin diglucosides: peonidin 3,5-O-diglucoside, malvidin 3,5-O-diglucoside, peonidin 3-O-(6' '-O-p-coumaroyl)-5-O-diglucoside, and malvidin 3-O-(6' '-O-p-coumaroyl)-5-O-diglucoside are the four major anthocyanins. The presence of pelargonidin 3-O-glucoside, not previously reported, has been established for the first time in all three juices.     

Time in lairage needed by pigs to recover from the stress of transport.

Two experiments were carried out in which a total of 602 pigs were slaughtered after being held in lairage for periods ranging from less than one hour to 21 hours. In the first experiment the pigs were handled under ideal conditions and slaughtered at the University of Bristol slaughterhouse; in the second the pigs were killed at a commercial plant. Blood samples collected at exsanguination were analysed for indices of stress. There were no consistent effects of time in lairage on the levels of lactate and creatine phosphokinase. Plasma cortisol and beta-endorphin levels were reduced by lairage for three hours or more in the first experiment and cortisol was reduced by lairage for two hours or more in the second; beta-endorphin was not measured in the second experiment. A period of rest in lairage allowed the pigs to recover from transport and the associated handling and the recovery appeared to be complete within two to three hours.     

Effect of paclobutrazol and salinity on ion leakage,proline content and activities of antioxidant enzymes in mango (Mangifera indica L.)

A study was conducted to investigate the effects of paclobutrazol (PBZ) on ion leakage (IL), proline content and activities of antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) of 1-year-old ‘Olour’ mango plants subjected to NaCl stress. Plants were treated with two levels of salt, i.e., 0.0 g NaCl (control) and 25 g NaCl/25 kg soil and three levels of paclobutrazol (PBZ) solution (0.0 (control), 750 and 1500 mg/l). Ion leakage, proline content and activities of antioxidant enzymes were significantly altered by both salinity and PBZ treatments. Results indicated that PBZ (1500 mg/l) mitigated the salinity stress and reduced ion leakage of mango seedlings by 64% over non-PBZ-treated salinised plants. PBZ upregulated the endogenous proline content and salinised plants treated with PBZ (1500 mg/l) had 17% higher proline content than salinised plants without PBZ treatment. Higher antioxidant enzyme activity was also observed in salinised plants treated by PBZ than salinised plants without PBZ treatments. Moreover, higher dose of PBZ (1500 mg/l) resulted in higher activity of these enzymes in mango leaves. In comparison to salinised plants without PBZ treatment, salinised plants treated with PBZ (1500 mg/l) had higher SOD (24%), CAT (46%) and POD (163%) activities. Our results suggest that PBZ application under salt stress conditions alters the equilibrium between free radical production and enzymatic defense reactions in mango by enhancing the proline content and free radical scavenging capacity.     

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.     

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.     

Saponins from Styrax officinalis

Three triterpene saponins named Styrax-saponin A-C (1-3) were found in pericarps of Styrax officinalis together with the deacylsaponin (4). Structural determinations were achieved using 1D-, 2D-NMR and mass spectrometry.     

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.     

The first serological evidence for Rift Valley fever infection in the camel,goitered gazelle and Anatolian water buffaloes in Turkey

Tropical Animal Health and Production - Rift valley fever (RVF), a vector-borne zoonotic disease, is caused by a phlebovirus (family Bunyaviridae). The virus was initially characterized...