Evaluation of polyherbal feed supplement (Ban-V) on the enhancement of growth and non-specific immune responses in shrimp Penaeus vannamei

Owing to the growing need for eco-friendly aquaculture, usage of herbal feed supplement receives higher attention. A commercial shrimp feed supplement known as Ban-V is a unique blend of polyherbal extracts that act synergistically with each other to help in improving the immune system, protects hepatopancreas and activates several metabolic pathways for better growth and disease resistance. For that, four supplemental concentrations such as 0.5, 1.0, 2.0 and 3.0 % of Ban-V were used along with commercial shrimp feed, and feed with no supplement (control) were evaluated for 60 days. Results revealed that 2.0% of Ban-V diet fed group showed significantly increased biological performance like (average body weight, average daily growth, feed conversion ratio and survival rate) than control. The tested immunological parameters such as total haemocyte count, oxyhaemocyanin level, phenoloxidase (PO) activity, respiratory burst activity (RB), superoxide dismutase (SOD), lysozyme activity, plasma protein content (PPC) and bactericidal activity were higher in 2.0% test diet fed P. vannamei, when compared to other test and control diet fed shrimp. It is therefore suggested that Ban-V supplementation at a concentration of 2.0% was efficient in stimulating the growth and immunity in shrimp.

     

Shallow Hydrothermal Vent Bacteria and Their Secondary Metabolites with a Particular Focus on Bacillus

Extreme environments are hostile for most organisms, but such habitats represent suitable settings to be inhabited by specialized microorganisms. A marine shallow-water hydrothermal vent field is located offshore in northeast Taiwan, near the shallow shore of the southeast of Kueishantao Island (121°55′ E, 24°50′ N). Research on extremophilic microorganisms makes use of the biotechnological potential associated with such microorganisms and their cellular products. With the notion that extremophiles are capable of surviving in extreme environments, it is assumed that their metabolites are adapted to function optimally under such conditions. As extremophiles, they need specific culture conditions, and only a fraction of species from the original samples are recovered in culture. We used different non-selective and selective media to isolate bacterial species associated with the hydrothermal vent crab Xenograpsus testudinatus and the sediments of its habitat. The highest number of colonies was obtained from Zobell marine agar plates with an overall number of 29 genetically distinct isolates. 16S RNA gene sequencing using the Sanger sequencing method revealed that most of the bacterial species belonged to the phylum Firmicutes and the class Bacilli. The present study indicates that hydrothermal vent bacteria and their secondary metabolites may play an important role for the reconstruction of the evolutionary history of the phylum Procaryota.     

Physiological effect of chitinase purified from Bacillus subtilis against the tobacco cutworm Spodoptera litura Fab.

An extracellular chitinase was purified from Bacillus subtilis. The lethal concentration (LC₅₀) was determined by using chitinase in first, second, and third instars of Spodoptera litura Fab. Chitinase showed the highest insecticidal activity at 6 μM concentration within 48 h. The nutritional indices were also significantly affected by the 6 μM concentration (P < 0.05). Food consumption, efficiency of conversion of ingested and digested food, relative growth rate, and consumption values declined significantly while approximate digestibility was increased. Our study indicates that treatment of host plant leaves with the chitinase can regulate (reduce) larval growth and weight, and enhance the mortality. This may serve as an effective biocide and alternative to Bt toxin.     

Influence of N fertilisation on the isolation of Acetobacter diazotrophicus and Herbaspirillum spp. from Indian sugarcane varieties

 Observations of newer diazotrophs like Acetobacter diazotrophicus and Herbaspirillum spp. in Indian sugarcane varieties is reported here for the first time. A. diazotrophicus was isolated from sugarcane plants where N fertilisation was nil, and numbers in the Indian cultivars were estimated to be up to 10⁶–10⁷ colony forming units g¹ fresh weight. N fertilisation was the limiting factor for the isolation of A. diazotrophicus, as evidenced by selective growth patterns in different concentrations of N sources, but Herbaspirillum spp. were found to occur both in N fertilised and unfertilised samples. The fact that N is not the limiting factor for Herbaspirillum spp. was evidenced in a similar study of its growth pattern in different N sources. Received: 14 May 1998     

Molecular cloning,sequence analysis and tissue expression of translationally controlled tumour protein from the WSSV‐infected Indian shrimp Penaeus indicus

The gene coding for translationally controlled tumour protein (TCTP) was polymerase chain reaction amplified from haemocyte cDNA of Indian shrimp, Penaeus indicus, and sequenced. The N‐terminal region, a conserved one among all the TCTPs, was shown to have one substitution at position 37, in the Indian isolate. Besides this, there were two substitutions in the C‐terminal region (135, 149), exclusive to the Indian isolate. Phylogenetic analysis suggested a close relatedness of TCTP from P. indicus to Fenneropenaeus chinensis compared with other isolates. Translationally controlled tumour protein gene expression was found to be elevated in the haemocytes of WSSV‐infected shrimps compared with the uninfected ones. However, tissues from the infected shrimps did not exhibit any detectable levels of TCTP expression.     

Improving hybrid rice parental lines for blast resistance by introgression of broad-spectrum resistance genes Pi54 and Pi9 by marker-assisted selection

Hybrid rice technology offers a great promise to produce 15% to 20% more yield than pure line varieties. The success of hybrid rice hinges on developing superior parental lines. To improve the blast resistance of hybrid rice parental line RP5933-1-19-2R, crosses were made with donors of two major blast resistance genes namely, Pi54 (Tetep) and Pi9 (IR71033–121-15) and the resulting F₁s were confirmed for their hybridity by using Pi54MAS and NMSMPi9-1 genic markers. The confirmed F₁s were intercrossed to obtain ICF₁s and selected positive plants by markers were backcrossed to the recurrent parent, as well as selfed for advancing further to BC₁F₃ and ICF₄ generations. The segregating plants were phenotyped for blast resistance at Uniform Blast Nursery. The identified complete restorers namely, RP 6619-1, RP 6616-26, RP 6619-3 and RP 6619-11 with Pi9 and Pi54 genes would serve as donors for broad spectrum blast resistance. This could ultimately lead to the development of new rice hybrids with improved resistance to blast disease, which is crucial for sustainable rice production and food security.     

In vitro lipopolysaccharide treatment alters regulatory T cell properties in chickens

The objective of this study was to identify the effect of in vitro lipopolysaccharide (LPS) treatment on regulatory T cells (Tregs) from chickens. Tregs had approximately 30-fold higher TLR 2-type 2 and six-fold higher TLR 4 mRNA content than CD4+CD25- cells. Tregs were treated with either 0 or 1 μg/ml LPS for 0, 2, and 4d. LPS treatment increased the IL-2 mRNA amount in Tregs at 2 and 4d post-LPS treatment. LPS treatment increased the IL-10 mRNA amount in Tregs at 4d post-LPS treatment. The total live cell numbers were approximately two-fold higher at 2d and three-fold higher at 4d in the 1 μg/ml LPS-treated groups than in the 0 μg LPS-treated controls. LPS treatment abrogated suppressive properties of Tregs at 2d post-LPS treatment. At 4d post-LPS treatment, Tregs became supersuppressive. In conclusion, chicken Tregs are differentially activated to facilitate immune response during the early stage of inflammation and to facilitate immune suppression at a later stage of inflammation.     

Effects of Bacillus subtilis metabolites on larval Aedes aegypti L

The culture supernatant of a strain of Bacillus subtilis isolated from soil samples killed larvae of the mosquito Aedes aegypti. The metabolites produced by B. subtilis were characterized using high performance liquid chromatography (HPLC). Mortality rate was dose-dependent for all larval instars of A. aegypti. Log probit analysis (95% confidence level) revealed an LC₅₀ of 1.73 and an LC₉₀ 3.71 μg/ml. Molecular weights/masses of B. subtilis metabolites were confirmed using SDS–PAGE analysis. B. subtilis metabolites were confirmed using HPLC analysis. We demonstrate that secondary metabolites from B. subtilis have larvicidal activity against A. aegypti and may be suitable for the control of this and other mosquito vectors of human disease. The larvae to the metabolites, significant reduction in the activities of acetylcholinesterse, α-carboxylesterase, and acid phosphatases were recorded.     

Regulatory T cell properties of thymic CD4(+)CD25(+) cells in ducks

Thymic CD4(+)CD25(+) cells from ducks were characterized for mammalian T regulatory cells' suppressive and cytokine production properties. The cross reactivity of anti-chicken CD25 monoclonal antibody with duck CD25 was confirmed by evaluating Concanavalin-A-stimulated CD25 upregulation in splenocytes. CD4(+)CD25(+) cells were detectable in the thymus, spleen, cecal tonsil, and lung (airsacs), but not in the bursa. Duck CD4(+)CD25(+) cells had approximately nine-fold higher IL-10 mRNA, 12-fold higher TGF-β, 16-fold higher CTLA-4, and nine-fold higher LAG-3 mRNA amounts than thymic CD4(+)CD25(-) cells. Thymic CD4(+)CD25(+) cells had no detectable levels of IL-2 mRNA. Duck CD4(+)CD25(+) cells had a three-fold higher IL-10 mRNA amount than chicken CD4(+)CD25(+) cells. Duck CD4(+)CD25(+) cells were anergic in vitro. Duck CD4(+)CD25(+) cells suppressed naive cell proliferation at effector: responder cell ratios above 0.5:1 in both contact-dependent and -independent pathways. It could be concluded that thymic CD4(+)CD25(+) cells in ducks are most likely the counterpart of mammalian T regulatory cells.