Effects of potential probiotic Bacillus subtilis KADR1 and its subcellular components on immune responses and disease resistance in Labeo rohita

The present study was conducted to determine the effects of Bacillus subtilisKADR1 and its subcellular components on immunity and disease resistance in Labeo rohita against Aeromonas hydrophila infection. Fish were fed diet containing different concentrations of live bacterial cells (DI‐10⁶, DII‐10⁸ and DIII‐10¹⁰ CFU/g) and another group of fish were immunized intraperitoneally with cellular components (WCPs, CWPs and ECPs) of Bacillus subtilisKADR1. After 4 weeks of trial, fish were challenged intraperitoneally with Aeromonas hydrophila cell suspension and survival percentage was recorded. Significantly higher post‐challenge survivability was recorded in fish groups fed 10⁸ CFU/g of KADR1 (80.24%; RPS = 75.76%) or immunized with WCPs (77.77%; RPS = 72.73%), compared with the control (18.51%). Analysis of immunological parameters viz. serum lysozyme, phagocytosis, serum total protein, respiratory burst, serum IgM levels, superoxide dismutase and alternative complement pathway activity reflected significant enhancement (p < .05) of immune response in fish fed 10⁶, 10⁸ and 10¹⁰ CFU/g of live cells, or immunized with cellular components of Bacillus subtilisKADR1, with the highest values were observed DII fed group, followed by the group immunized with WCPs. Our results suggest that dietary administration of Bacillus subtilisKADR1 at 10⁸ CFU/g can effectively enhance the immune responses and disease resistance in Labeo rohita against Aeromonas hydrophila.     

Stress-strain curves and corresponding structural parameters in mulberry and non-mulberry silk fibers

Mulberry fiber (Bivoltine) and non-mulberry fiber (Tassar) were subjected to stress-strain studies and the corresponding samples were examined using wide angle X-ray scattering studies. Here we have two different characteristic stress-strain curves and this has been correlated with changes in crystallite shape ellipsoids in all the fibers. Exclusive crystal structure studies of Tassar fibers show interesting feature of transformation from antiparallel chains to parallel chains.     

Comparative Analysis of CD4+ and CD8+ T Cells in Tumor Tissues,Lymph Nodes and the Peripheral Blood from Patients with Breast Cancer

Background: CD4+ and CD8+ T cells are the main types of lymphocytes in cell-mediated immunity and play a central role in the induction of efficient immune responses against tumors. The frequencies of T cell subtypes in the peripheral blood and tumor tissues, and draining lymph nodes (dLN) can be considered as useful markers for evaluation of the immune system in cancers. Methods: In this study, the frequencies of CD4+ and CD8+ T cells in blood, tumor tissues, and dLN samples of breast cancer patients were compared with each other and with similar tissues from normal individuals. Immunophenotyping was carried out by flow cytometry and the expression levels of CXCL10, granzyme B, and mammaglobin were evaluated by real-time PCR. Results: In the peripheral blood, there were no differences in the T cell subsets between the patients and the normal individuals. The frequency of CD8+ T cells was significantly higher in tumor tissue than normal breast tissues while granzyme B expression was similar. Based on mammaglobin expression levels, dLN have been classified into micro- and macro-metastatic dLN. We found significantly lower frequency of CD4+ in macro-metastatic dLN than micro-metastatic dLN. CD8+ frequency was similar in both dLN; however, granzyme B expression was higher in micro-metastatic ones. There was not any significant difference in CXCL10 expression between the two types of dLN. Conclusion: Based on our results, although the tumor does not affect the systemic immunity, tumoral cells affect the local immune system in the tumoral tissues and the metastatic dLN. Key Words: Breast neoplasms, CD4+ T lymphocyte, CD8+ T lymphocytes, CXCL10, Granzymes     

Bio-potency of serine proteinase inhibitors from Acacia senegal seeds on digestive proteinases, larval growth and development of Helicoverpa armigera (Hübner)

Proteinase inhibitors (AsPIs) with high activity against serine proteinases were purified from seeds of the tree legume, Acacia senegal by ammonium sulfate precipitation followed by DEAE-Sephadex A-25 column and evaluated against Helicoverpa armigera larvae by in vitro and in vivo methods. The molecular weight of AsPIs was found to be approximately 19.58 ± 1.00 and 21.23 ± 1.00 kDa for PI and 18.16 ± 1.00 kDa for PII on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The AsPIs (5 μg/ml) inhibited approximately 70% of midgut trypsin and 61% of elastase-like chymotrypsin. In vitro studies showed that AsPIs have remarkable inhibitory activity towards total gut proteolytic enzymes followed by trypsin and chymotrypsin. The IC₅₀ of AsPIs for midgut trypsin was 0.1 μg/ml and for chymotrypsin was 2.0 μg/ml. The inhibition of gut proteinase enzymes was of the non-competitive type. In larval feeding studies, AsPIs were found to retard growth and development of H. armigera and also affects the fecundity of the pest. The results advocate the use of AsPIs in transgenic technology to develop plant resistance to H. armigera.     

Impacts of Bioinoculants Pseudomonas jesenii MP1 and Rhodococcus qingshengii S10107 on Chickpea (Cicer arietinum L.) Yield and Soil Nitrogen Status

Cold-adapted bioinoculants are considered as harbingers of sustainable hill agriculture. Therefore, two previously characterized psychrotolerant diazotrophs, Pseudomonas jesenii MP1 and Rhodococcus qingshengii S10107, were evaluated for their plant growthpromoting potential for chickpea (Cicer arietinum L.) grown under natural field conditions. Comparative analysis of agronomical and biochemical crop parameters revealed the irrelevance of chemical fertilizers for chickpea production; the diazotrophs alone were sufficient to fulfil the crop''s nutritional requirement. However, the integrated use of bacterial strains in combination with urea at 20 kg N ha^-1 as urea was being recommended for higher crop yield and better soil nitrogen status. Quantitative polymerase chain reaction (qPCR) and denaturing gradient gel electrophoresis (DGGE)-based soil bacterial dynamics unveiled the persistence of both diazotrophs until the end of the crop maturation period without affecting the native micro-flora. Therefore, these bioinoculants can be explored as natural nitrogen resource, and an additional incentive in their bio-formulation will be a step towards agricultural sustainability.     

Impacts of elemental S applied under various temperature and moisture regimes on pH and available P in acidic, neutral and alkaline soils

We evaluated the effect of elemental S (S⁰) under three moisture (40, 60, 120% water-filled pore space; WFPS) and three temperature regimes (12, 24, 36°C) on changes in pH and available P (0.5 N NaHCO₃-extractable P) concentrations in acidic (pH 4.9), neutral (pH 7.1) and alkaline (pH 10.2) soils. Repacked soil cores were incubated for 0, 14, 28 and 42 days. Application of S⁰ did not alter the trends of pH in acidic and neutral soils at all moisture regimes but promoted a decrease in the pH of alkaline soil under aerobic conditions (40%, 60% WFPS). Moisture and temperature had profound effects on the available P concentrations in all three soils, accumulation of available P being greatest under flooded conditions (120% WFPS) at 36°C. Application of S⁰ in acidic, neutral and alkaline soils resulted in the net accumulation of 16.5, 14.5 and 13 g P g^–1 soil after 42 days at 60% WFPS, but had no effect under flooded conditions. The greatest available P accumulations in the respective soils were 19, 19.5 and 20 g P g^–1 soil (equivalent to 38, 41, 45 kg P ha^–1) with the combined effects of 36°C, 60% WFPS and applied S⁰. The results of our study revealed that oxidation of S⁰ lowered the pH of alkaline soil (r=–0.88, P<0.01), which in turn enhanced available P concentrations. Also, considering the significant relationship between the release of sulphate and accumulation of P, even in acidic soil (r=0.92, P<0.01) and neutral soil (r=0.85, P<0.01) where the decrease in pH was smaller, it is possible that the stimulatory effect of sulphate on the availability of P was due to its concurrent desorption from the colloidal surface, release from fixation sites and/or mineralization of organic P. Thus, in the humid tropics and irrigated subtropics where high moisture and temperature regimes are prevalent, the application of S⁰ could be beneficial not only in alleviating S deficiency in soils but also for enhancing the availability of P in arable soils, irrespective of their initial pH.     

Acrylamide in French fries: influence of free amino acids and sugars

The free amino acid profile and sugar (fructose, glucose, and sucrose) composition were determined in potato samples selected to give a large range of variation (a total of 66 samples). From these samples French fries were produced in a laboratory-scale simulation of an industrial process followed by a finish fry at 180 degrees C for 3.5 min using a restaurant fryer. The final product was blast frozen and analyzed for acrylamide. Acrylamide was detected in all samples, but its concentration varied significantly from 50 to 1800 ng/g. For isotope dilution (13C3) acrylamide analysis, samples were extracted with water, cleaned up on HLB Oasis polymeric and Accucat mixed mode anion and cation exchange SPE columns, and analyzed by LC-MS/MS. Statistical analysis of the data indicates that the effect of sugars and asparagine on the concentration of acrylamide in French fries is positive and significant (p < 0.001). It appears that one of the ways acrylamide formation in French fries can be effectively controlled is by the use of raw products with low sugar (and to a lesser degree, asparagine) content.     

Polyclonal antibody‐based farmer‐friendly flow‐through test for the detection of acute hepatopancreatic necrosis disease in shrimp

Early mortality syndrome (EMS) or acute hepatopancreatic necrosis disease (AHPND) is currently the most significant disease of shrimp in farms of Vietnam, Thailand, Malaysia, China and Mexico, and there is a great risk that it may spread to other shrimp farming countries. Although, an array of sophisticated detection tools for AHPND available, there is a need for a sensitive, simple and rapid detection method. In this study, a simple, sensitive, rapid and polyclonal antibody‐based farmer‐friendly flow‐through assay (FTA) test has been developed for the detection of AHPND pathogen. The recombinant Photorhabdus insect‐related (Pir) A toxin‐like protein of AHPND pathogen was used to immunize rabbits at 21‐day interval observed for highest antibody titre after third booster by ELISA. The raised rabbit antiserum was purified by affinity chromatography and characterized by Western blot. The antiserum showed no cross‐reactivity with AHPND‐free Vibrio parahaemolyticus, V. anguillarum, White Spot Virus (WSV), Aeromonashydrophila and Aphanomycesinvadans. This polyclonal rabbit antiserum was used to develop a farmer‐friendly FTA test for the detection of AHPND pathogen. This simple FTA testis is more sensitive and could detect PirA^VP toxin up to 0.121 µg/ml, compared with 0.242 µg/ml by immunodot assay. Furthermore, FTA test requires only 8–10 min for completion, compared with 3 hr by immunodot thus found to be more sensitive, specific and cost‐effective. Collectively, sensitive FTA test would help shrimp farmers to take real‐time management decisions, especially emergency harvest and finally be a better hope for the prevention of AHPND.