Purification and Characterization of a Novel Alginate Lyase from a Marine Streptomyces Species Isolated from Seaweed

Alginate, a natural polysaccharide derived from brown seaweed, is finding multiple applications in biomedicine via its transformation through chemical, physical, and, increasingly, enzymatic processes. In this study a novel alginate lyase, AlyDS44, was purified and characterized from a marine actinobacterium, Streptomyces luridiscabiei, which was isolated from decomposing seaweed. The purified enzyme had a specific activity of 108.6 U/mg, with a molecular weight of 28.6 kDa, and was composed of 260 amino acid residues. AlyDS44 is a bifunctional alginate lyase, active on both polyguluronate and polymannuronate, though it preferentially degrades polyguluronate. The optimal pH of this enzyme is 8.5 and the optimal temperature is 45 °C. It is a salt-tolerant alginate lyase with an optimal activity at 0.6 M NaCl. Metal ions Mn²⁺, Co²⁺, and Fe²⁺ increased the alginate degrading activity, but it was inhibited in the presence of Zn²⁺ and Cu²⁺. The highly conserved regions of its amino acid sequences indicated that AlyDS44 belongs to the polysaccharide lyase family 7. The main breakdown products of the enzyme on alginate were disaccharides, trisaccharides, and tetrasaccharides, which demonstrated that this enzyme acted as an endo-type alginate lyase. AlyDS44 is a novel enzyme, with the potential for efficient production of alginate oligosaccharides with low degrees of polymerization.     

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.     

Picobirnavirus detection in bovine and buffalo calves from foothills of Himalaya and Central India

The present study describes detection of picobirnavirus (PBV) in faecal samples from bovine and buffalo calves employing the polyacrylamide gel electrophoresis (PAGE). A total of 136 faecal samples from buffalo (n = 122) and cow calves (n = 14) exhibiting clinical signs of diarrhoea and from healthy calves were collected during 2007–2010 from subtropical (central India) and tarai area of western temperate Himalayan foothills (Uttarakhand). The dsRNA nature of the virus was confirmed by nuclease treatment (RNase A, RNaseT1 and DNase 1). PAGE results confirmed 3.67% (5/136) positivity for PBV, showing a typical genomic migration pattern with two discrete bands with size of approximately 2.4 and 1.7 kbps for the larger and smaller segments, respectively. Among the five PBV samples identified, three were from buffalo calves and one from cow calf exhibiting clinical signs of acute diarrhoea, while one sample from non-diarrhoeic buffalo calf also showed the presence of PBV. None of the samples showed dual infection of rotavirus and PBV. The preliminary findings indicate sporadic incidences of PBV in bovine calves and emphasize the need for the development of better diagnostics for early detection and genetic characterization of these emerging isolates of farm animals of economic significance.     

Pharmacokinetics and physio‐metabolic response of single and multiple dose of fenbendazole in Labeo rohita (Hamilton, 1822) fingerlings

An experiment was carried to determine the plasma fenbendazole (FBZ) concentration and physio‐metabolic responses in juveniles of Labeo rohita (90 ± 4 g) after oral administration of single doses at 10, 20 and 50 mg, 20 mg FBZ/kg b.wt. in multiple times on 1st, 3rd and 7th day. The blood samples were collected at 0.5, 1, 2, 4, 8, 12, 24, 30, 48, 72, 96 and 120 hr, after single‐dose administration, and regularly (upto 15 day) in multiple dose. Plasma FBZ concentration was determined up to the limit of detection (LoD) of 0.09 µg/ml by HPLC. There was no parent drug detected in plasma for administration of 10 mg FBZ/kg b.wt. The drug attained the peak concentration (C_max) 1.85 and 3.09 µg/ml in plasma at 4 hr (T_max) after administration of 20 and 50mg FBZ/kg b.wt. respectively. Plasma FBZ was detectable up to 96 and 120 hr with concentration 0.09 ± 0.007 and 0.098 ± 0.006 µg/ml, respectively, after single‐dose administration of 20 and 50mg/kg b.wt. In case of multiple‐dose administration, the maximum concentration of FBZ was 1.01 ± 0.03 µg/ml on 7th day that was less than to the single dose at 50 mg/kg b.wt. However, FBZ was detected up to 11 day after multiple doses. The study revealed that the hepatic antioxidant enzymes activities like superoxide dismutase, catalase and glutathione‐S‐transferase were significantly affected by increasing FBZ in single and multiple doses. The results of the present study could reveal that single‐ or multiple‐oral administration of FBZ at 20 mg/kg b.wt. in feed as antihelminthic drug in L. rohita could be considered as the safe dose.     

Optimization of de‐oiled rice bran inclusion level in the diet of Labeo rohita: Effect on growth performance,nutrient utilization,insulin‐like growth factors I and II gene expression

A 60‐day feeding trial was conducted to optimize the inclusion level of de‐oiled rice bran (DORB) in the diet of Labeo rohita fingerlings. Six iso‐nitrogenous (260g/kg), iso‐lipidic (70g/kg) and iso‐energetic (15 MJ/Kg) diets were prepared with different inclusion level of DORB viz., T33, T38_, T43, T48, T53 and T58. One hundred and eighty fingerlings (9.8 ± 0.5 g) were randomly distributed in seven experimental groups in triplicates following a completely randomized design. The weight gain %, specific growth rate, protein efficiency ratio and feed efficiency ratio were found similar in 33%, 38% and 43% DORB fed groups. The expression of both insulin‐like growth factors I and II (IGF‐I and II) was significantly (p < 0.05) higher in T33, T38 and T43 groups. Based on second order polynomial regression analysis (y = ?0.0059x² + 0.4573x ? 4.8404, R² = 0.95), the expression of IGF‐I was found to be maximum at 39.5% inclusion level of DORB. Significantly (p < 0.05) lower muscle RNA content, RNA‐DNA ratio and higher feed conversion ratio were found in T58 group. Hence, from these results, it can be concluded that the optimum inclusion of DORB in the diet (260g/kg CP and 70g/kg lipid) of L. rohita should be 39.5% to support maximum nutrient utilization and growth performance.     

Effect of a decade-long chemical fertilizers and amendments application on potassium fractions and yield of maize​–wheat in an acid Alfisol

Effect of long-term addition of chemical fertilizers with or without amendments was studied on different forms of potassium and the yield of maize and wheat. Continuous application of chemical fertilizers and amendments for 40 years influenced different fractions of potassium significantly. Integrated use of a balanced dose of chemical fertilizer, with farmyard manure (FYM) or lime, sustained higher yields of maize and wheat in comparison to inorganic fertilizers alone. Application of urea (100%) N alone for 40 years resulted in zero yield level. Continuous application of chemical fertilizers either alone or in combination with FYM or lime influenced different fractions of potassium significantly. Continuous cropping without fertilization resulted in depletion to the order of 21.5%, 16.6%, 11.7%, and 5.5% in water-soluble, exchangeable, 0.5 N HCl extractable, and non-exchangeable K, respectively. Different fractions of potassium were found to be positively and significantly correlated with grain and stover/straw yield of maize and wheat.     

Comparison of Cell Disruption Methods for Improving Lipid Extraction from Thraustochytrid Strains

Lipid extraction is an integral part of biodiesel production, as it facilitates the release of fatty acids from algal cells. To utilise thraustochytrids as a potential source for lipid production. We evaluated the extraction efficiency of various solvents and solvent combinations for lipid extraction from Schizochytrium sp. S31 and Thraustochytrium sp. AMCQS5-5. The maximum lipid extraction yield was 22% using a chloroform:methanol ratio of 2:1. We compared various cell disruption methods to improve lipid extraction yields, including grinding with liquid nitrogen, bead vortexing, osmotic shock, water bath, sonication and shake mill. The highest lipid extraction yields were obtained using osmotic shock and 48.7% from Schizochytrium sp. S31 and 29.1% from Thraustochytrium sp. AMCQS5-5. Saturated and monounsaturated fatty acid contents were more than 60% in Schizochytrium sp. S31 which suggests their suitability for biodiesel production.     

Effect of manures and inorganic fertilizers on growth,yield, and essential oil of damask rose (Rosa damascena Mill.) and chemical properties of soil in western Himalayas

A field experiment was conducted to study different manures and inorganic fertilizers effect on damask rose during 2008–2012. The rose essential oil (EO) was obtained by hydro distillation in Clevenger type apparatus and the components in the rose oil were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). Application of 90:80:90 kg nitrogen: phosphorus: potassium (NPK) ha^?1 outshine all other treatments and recorded significantly higher flower and oil yield as compared to control. EO yield was also significantly correlated with flower yield plant^?1, number of flowers plant,^?1 and flower yield ha^?1. The higher percentage of citronellol+nerol was obtained by the application of 120:40:90 kg NPK ha^?1. Geraniol (26.2%) was maximum when the plants were fertilized with 90:40:90 kg NPK ha^?1. Citronellol+nerol/geraniol ratio was also high in fertilized plots. Furthermore, manure application resulted in a marked improvement in chemical properties (pH, percent organic carbon, available N, P, and K) of the soil.