In Vitro Characterization of <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> Strains with Inhibitory Activity on Enteropathogens for Use as Potential Animal Probiotics

The present study evaluates the probiotic properties of three Lactobacillus plantarum strains MJM60319, MJM60298, and MJM60399 possessing antimicrobial activity against animal enteric pathogens. The three strains did not show bioamine production, mucinolytic and hemolytic activity and were susceptible to common antibiotics. The L. plantarum strains survived well in the simulated orogastrointestinal transit condition and showed adherence to Caco-2 cells in vitro. The L. plantarum strains showed strong antimicrobial activity against enterotoxigenic Escherichia coli, Shiga toxin-producing E. coli, Salmonella enterica subsp. enterica serovar Typhimurium, Choleraesuis and Gallinarum compared to the commercial probiotic strain Lactobacillus rhamnosus GG. The mechanism of antimicrobial activity of the L. plantarum strains appeared to be by the production of lactic acid. Furthermore, the L. plantarum strains tolerated freeze-drying and maintained higher viability in the presence of cryoprotectants than without cryoprotectants. Finally, the three L. plantarum strains tolerated NaCl up to 8% and maintained >60% growth. These characteristics of the three L. plantarum strains indicate that they could be applied as animal probiotic after appropriate in vivo studies.     

Optimal analytical conditions for catalase in fresh water prawn, Macrobrachium malcolmsonii

The cytosol of hepatopancreas was prepared from the freshwater prawn Macrobrachium malcolmsonii, and optimal assay conditions, i.e., concentration of substrate, pH, and temperature, were determined to measure basal activities and kinetic constants of catalase activity. The properties of catalase were examined in M. macolmsonii, because quantitative data on catalase are limited in crustacean species. The optimal pH for catalase was 7.0. The activation energy was 3.55 Kcal/mol and energy inhibition value was 5.16 Kcal/mol. The value of energy inhibition is higher than that of energy activation. This may be due to inhibition of catalase by some substrate other than H2O2. A Km of 66.6 mM was also determined from various concentrations of substrate.     

Ecosystem Restoration of Jhum Fallows in Northeast India: Microbial C and N Along Altitudinal and Successional Gradients

Abstract To characterize the altitudinal and successional trends in microbial biomass and to understand their role in soil nutrient dynamics during the aggradation phase (vegetation recovery) of abandoned shifting cultivation systems, we determined the soil properties and microbial C and N in jhum (slash‐and‐burn) cultivation systems at different altitudes and 1‐, 7‐, and 16‐year‐old fallow agricultural lands at lower and higher altitudes in the northeastern Indian hills. Density of ground vegetation was lower in the undisturbed forest than in the jhum fallows. In general, 1‐year jhum fallow had greater herbaceous vegetation both at lower and higher altitudes. Although woody plants were observed in 7‐ and 16‐year‐old jhum fallows, their density was highest in the forest. Soil moisture, organic C, and total N also increased gradually with increasing altitude and progressive secondary succession. Soil pH showed a negative correlation with altitude (as also confounded by soil type) and fallow age. Both microbial C and N had a close correlation with altitude and fallow age. Contribution of microbial C to soil organic C was 2.0–2.6% and microbial N to total N 1.4–2.2% in jhum fields, 2.4–4.3% and 1.2–2.1%, respectively, in jhum fallows, and 2.5–2.9% and 1.6–1.9% in the forests. Microbial C and N showed a negative correlation with herbaceous plant density. Microbial biomass in the jhum fallows and forest stands had a positive relationship with woody vegetation. Along an altitudinal and/or successional gradient, microbial C and N were positively correlated with water‐holding capacity, soil moisture, organic C, and total N and negatively correlated with soil pH. Microbial C and N were positively correlated with each other. Therefore, the study suggests that the altitudinal and successional dynamics of microbial C and N are linked to, among other properties, soil organic matter and total nitrogen contents in the soil during community development after land abandonment from shifting cultivation.     

Cloning of sarco-endoplasmic reticulum Ca<Superscript>2+</Superscript>-ATPase (SERCA) from Caribbean spiny lobster <Emphasis Type="Italic">Panulirus argus</Emphasis>

We have previously reported on calcium transport mechanisms in American lobster, Homarus americanus, using ⁴⁵Ca²⁺ coupled with vesicle preparations of hepatopancreatic endoplasmic reticulum. The active transport of calcium across membranes bordering calcium-sequestering stores such as sarcoplasmic or endoplasmic reticulum is catalyzed by membrane-spanning proteins, the sarco-endoplasmic Ca²⁺-ATPases (SERCAs). In the study described here we used advanced bioinformatics and molecular techniques to clone SERCA from the economically important Caribbean spiny lobster, Panulirus argus. We report the complete cloning of a full-length SERCA from P. argus antenna cDNA (GenBank accession number AY702617). This cDNA has a 1020-amino acid residue open reading frame which is 90% identical to published sequences of other crustacean SERCA proteins. Our data support the hypothesis that one crustacean and three vertebrate genes controlling calcium transport were derived from a common ancestral gene.     

Relevance of Non-ceruloplasmin Copper to Oxidative Stress in Patients with Hepatocellular Carcinoma

Altered copper homeostasis and oxidative stress have been observed in patients with hepatocellular carcinoma. Non-ceruloplasmin copper, the free form, is a potent pro-oxidant than the protein bound copper. The aim of the present study was to evaluate which form of copper can be correlated with the oxidative stress in the circulation and in the malignant liver tissues of hepatocellular carcinoma patients. Hepatocellular carcinoma patients (grades II and III, n = 18) were enrolled in this study. Serum levels of total, free and bound copper, ceruloplasmin, iron, iron-binding capacity, lipid peroxidation products, and enzymatic and non-enzymatic antioxidants were quantified in serum and in malignant liver tissues and compared with those of normal samples (n = 20). A significant positive correlation between the serum non-ceruloplasmin copper and lipid peroxidation products and negative correlation with antioxidants were observed in hepatocellular carcinoma patients. In liver tissue, glutathione peroxidase, superoxide dismutase, and catalase activity were significantly decreased with concomitant elevation in oxidative stress markers. Our experiment revealed that the elevation in non-ceruloplasmin copper has high relevance with the oxidative stress than the bound copper.     

Association of CRISP2, CCT8, PEBP1 mRNA abundance in sperm and sire conception rate in Holstein bulls

The objective was to determine the association of mRNA expression of cystine rich secretary protein 2 (CRISP2), chaperonin containing T-complex protein 1, subunit 8 (CCT8), and phosphatidylethanolamine-binding protein 1 (PEBP1), in sperm of Holstein bulls with Sire Conception Rate (SCR) scores between −4 and +4. These proteins were involved in sperm capacitation and sperm-egg fusion. Samples of sperm obtained on a single day from Holstein bulls (N = 34) in a commercial AI centre were used to evaluate relative mRNA expression of CRISP2, CCT8, and PEBP1. The mRNA abundance of CRISP2 was positively correlated (r = 0.88; P < 0.002), CCT8 was negatively correlated (r = −0.87; P < 0.002), and PEBP1 was positively correlated (r = 0.83; P < 0.006) with SCR-scores. The means of CRISP2 mRNA abundance was greater among positive SCR-score bulls (2.5 to 8 fold), the means of CCT8 mRNA abundance was greater among the negative SCR-score bulls (9.5 to 3.5 fold), and the means of PEBP1 mRNA abundance was greater for the positive SCR-score bulls (5.4 to 7.7 fold). In multivariate regression models predicting SCR-scores, mRNA abundance of CCT8 was significantly associated with SCR-score in all models. In the presence of CRISP2 mRNA abundance in the model, the SCR score's predictability of PEBP1 was insignificant. However, in the absence of CRISP2 mRNA abundance in the model, the SCR-score's predictability of PEBP1 was significant. In multivariate regression models, CRISP2 and CCT8 mRNA expression in sperm accounted for 95% of the variance in Holstein bull's SCR-scores. In conclusion, Holstein bulls with greater CRISP2 and lower CCT8 mRNA expression in sperm had higher probabilities of siring calves.     

Ca2+-dependent activator protein for secretion 1 is critical for constitutive and regulated exocytosis but not for loading of transmitters into dense core vesicles

Although CAPS1 was originally identified as a soluble factor that reconstitutes Ca(2+)-dependent secretion from permeabilized neuroendocrine cells, its exact function in intact mammalian cells remains controversial. Here we investigate the role for CAPS1 by generating stable cell lines in which CAPS1 is strongly down-regulated. In these cells, Ca(2+)-dependent secretion was strongly reduced not only of catecholamine but also of a transfected neuropeptide. These secretion defects were rescued by infusion of CAPS1-containing brain cytosol or by transfection-mediated expression of CAPS1. Whole cell patch clamp recording revealed significant reductions in slow burst and sustained release components of exocytosis in the knockdown cells. Unexpectedly, they also accumulated higher amounts of endogenous and exogenous transmitters, which were attributable to reductions in constitutive secretion. Electron microscopy did not reveal abnormalities in the number or docking of dense core vesicles. Our results indicate that CAPS1 plays critical roles not only in Ca(2+)-dependent, regulated exocytosis but also in constitutive exocytosis downstream of vesicle docking. However, they do not support the role for CAPS1 in loading transmitters into dense core vesicles.     

Substrate selection and seasonal variation in densities of invertebrates in stream pools of a tropical river

Spatial distribution and seasonal variation in densities of the invertebrates were investigated for a year in three stream pools of a South Indian river. The effects of season, substrate type and water depth on the distribution were analyzed. Substrate type and season influenced the invertebrate distribution the most. Leaf packs harboured most of the organisms followed by macro-algal substrate and sand. The lowest densities were observed on rocky substrates and in the water column. Rocky substrates in shallow water supported higher densities of total invertebrates than deeper areas. Chironomid larvae dominated all benthic substrates throughout the year. Of the 19 invertebrate taxa studied, 6 showed no seasonality in densities, and most of the rest showed their highest densities in the pre-monsoon period and lowest in the SW monsoon or post-monsoon periods. However, in two of the three pools, the densities of total invertebrates were highest during the post-monsoon period with secondary peaks in the pre-monsoon period.     

Bovine interleukin 2: production kinetics in normal and in vivo antigen-primed cattle

Conditions influencing production kinetics of bovine interleukin 2 (IL-2), viz. cell concentration, mitogen and its concentration, length of incubation, nutrient medium and in vivo antigen-priming were investigated. Peripheral blood mononuclear cells (PBL) of outbred cattle of different age groups showed considerable variation in their ability to secrete IL-2 which possibly reflects their immune competence. Of the cultures initiated with PBL, 5 x 10(6) cells/ml cultured in serum free Iscove's medium and stimulated with 5 micrograms Con A/ml for 24 hr produced maximal amount of IL-2 activity. In vivo antigen-priming of bovine lymphocytes with the live attenuated rinderpest virus revealed that IL-2 production was not affected by rinderpest virus but the in vivo antigen-priming possibly resulted in concomitant production of suppressor factor(s) which suppressed the already produced IL-2. The implications of this factor(s) in relation to regulation of immune responses in the disease process are discussed.     

Feeding habit and food partitioning in a stream fish community of Western Ghats,India

The diet and feeding ecology of 12 cyprinid fishes were studied in the Thalayanai stream of southern India. The diet of Barbodes sarana, Danio aequipinnatus and Rasbora daniconius were specialized in terrestrial and aquatic insects. Hypselobarbus dobsoni and Garra mullya consumed vegetative matter and unicellular algae as their most important dietary component, respectively. Hypselobarbus curmuca and Hypselobarbus dubius specialized in crustaceans and molluscs. Puntius amphibious, Puntius bimaculatus, Puntius dorsalis, Puntius filamentosus and Puntius tambraparniei fed on variety of food resources (aquatic insects, algae and detritus) and their feeding intake varied between wet to dry seasons. Levin’s index of diet breadth showed the broadest diets for P. dorsalis in both seasons, whereas G. mullya had a narrow feeding spectrum in both seasons. Among co-existing species, diet overlaps were substantially low during both wet and dry season. These results suggest that feeding specialization and low diet overlap reduce inter-specific competition, allowing co-existence of these cyprinid fishes in this highly structured stream assemblage.