Comparison of dot blot and PCR diagnostic techniques for detection of white spot syndrome virus in different tissues of Penaeus monodon

Comparison of PCR and dot blot diagnostic techniques for detection of white spot syndrome virus (WSSV) was made on different tissues of infected Penaeus monodon including eye stalk, eye stalk with eye, gills, cuticle, pleopod, periopods, uropods and telson. Dot blots of crude DNA extracted from infected tissue samples showed positive reactions with all the samples; however, the sensitivity of the dot blot was reduced with the purification of DNA samples extracted from pleopod, telson and uropod. PCR was found to be more sensitive when compared to dot blot. Both crude DNA and purified DNA samples extracted from all the tissues except for eye stalk with eye showed single step nested PCR positive reaction. The amplification of all or either of the three bands of 941 bp, 525 bp and 204 bp size varied with the tissues analysed. The severity of infection assessed by PCR amplification was found to be maximum in cuticle and telson followed by gill. Other tissues such as eye stalk, pleopod, periopods and uropod were observed to have mild infection. The maximum intensity of the PCR product was for the smallest amplified product of 204 bp followed by 525 bp and the weakest intensity was observed for the 941 bp size. The limitation of PCR due to inhibiting factors present in tissues could be overcome with the use of dot blot which gave positive reaction from the DNA extracted from eye stalk including the eye but yielded no amplification by PCR.     

Antinociceptive activity of Wrightia tinctoria bark

The ethyl acetate, acetone and methanol extracts of Wrightia tinctoria bark showed antinociceptive activity on acetic acid-induced writhing test in mice, their effects being comparable to that of acetylsalicylic acid.     

Zero-water exchange shrimp farming systems (extensive) in the periphery of Chilka lagoon, Orissa, India

The extensive zero-water exchange shrimp farming system in the periphery of Chilka lagoon (Orissa, India) was studied. The study aimed to describe this unique farming system with special reference to dynamics of macrozoobenthos, production characteristics and economics. The study conducted was based on a general survey as well as monitoring of five individual farms over a complete production cycle. The farming practice in this area is characterized by complete absence of water exchange during rearing. Ponds in this area are generally shallow (mean 72 cm). Most of the water and soil quality characteristics of these farms are within acceptable levels. Macrozoobenthos belonging to 12 taxa were collected, amphipods (81%) and polychaetes (13%) being most numerous. Overall macrobenthic density of farms studied varied from 968 to 11,470 individuals/m² with a gross mean of 5644 individuals/m². There was no general pattern to the variation in abundance of various taxa in different phases of the rearing cycle, suggesting a low predatory pressure by shrimp in the farms studied. Shrimp production was highly variable (91–250 kg/ha), but generally low with a mean of 145 kg/ha. The net income of these farms was estimated to be Rs. 63,250 per crop per ha. Compared with shrimp farming system with regular water exchange in the same area, Chilka farms generated high benefit-cost ratio indicating high profitability and sustainability.     

Effects of C:N ratio and substrate integration on periphyton biomass, microbial dynamics and growth of Penaeus monodon juveniles

An outdoor growth trial was conducted for 75 days to investigate the effect of C:N ratio and bamboo substrate (S) in brackish water shrimp culture. Penaeus monodon juveniles (0.35 ± 0.03 g) were stocked in experimental tanks with and without bamboo substrate. C:N ratio of 10 and 20 was manipulated with shrimp feed containing 32 % crude protein and rice flour as carbohydrate source. Addition of substrate and higher C:N ratio (CN20+S) significantly reduced (p < 0.01) inorganic nitrogen NH₃–N by 48.2 %, NO₃–N by 41.6 %, NO₂–N by 42.7 % compared with CN10. Substrate addition significantly (p < 0.05) reduced the turbidity by 22.4 % (CN10+S against CN10) and 20.7 % (CN20+S against CN20). Periphyton biomass and total heterotrophic bacterial load were significantly higher (p < 0.05) in CN20+S in comparison with CN10+S treatment. The autotrophic index increased from 123.4 ± 0.62 to 158.9 ± 3.8 for CN10+S and, 121.8 ± 1.6 to 228 ± 9.11 for CN20+S system. Provision of substrate and carbohydrate addition resulted in the highest body weight, 4.87 ± 0.12 g in CN20+S against 3.66 ± 0.07 g in CN20 and 2.90 ± 0.12 g in CN10 through provision of natural food in the form of periphytic algae and heterotrophic bacterial community. Treatment with substrates showed a higher survival rate by 7 % in comparison with without substrate treatments as it acted as shelter to shrimp during molting and there by reduced cannibalism. Thus, our results demonstrated that high C:N ratio and substrate addition improved growth, reduced FCR and better water quality conditions.     

Selection of monoploids for protoplast fusion and generation of intermonoploid somatic hybrids of potato

The union of potato monoploid genotypes (2n=1x =12) through protoplast fusion may result in vigorous somatic hybrids due to a reduction of the “genetic load” normally present in this highly heterozygous tetraploid (2n=4x=48) crop. More than 100 androgenic monoploids derived from diploid (2n=2x=24)Solanum phureja Juz. & Buk. andS. chacoense Bitt. xS. phureja clones were evaluated in field trials during 1996 and 1997 to identify the most promising genotypes for protoplast fusion experiments. Compared to the total population, the 1996 selected genotypes had higher means for tuber number (30.1 vs 11.2 tubers/plant), average tuber weight (3.0 vs 1.8 g/tuber) and total yield (66.1 vs 20.4 g/plant). Similarly, the 1997 selected genotypes had higher means for tuber number (42.8 vs 25.4 tubers/plant), average tuber weight (3.6 vs 2.5 g/tuber) and total yield (114.0 vs 63.4 g/plant) compared to the total population. The 31 selected monoploid genotypes from 1996-97 varied in their response to protoplast isolation and culture from no growth (9), cell enlargement (5), limited cellular divisions (8), callus formation (5) to plant regeneration from callus (4). Chemical fusion and electrofusion produced three groups of intermonoploid somatic hybrids. Polymorphic simple sequence repeat (SSR) loci enabled distinction of somatic hybrids from parental somaclones. Rapid DNA extraction with SSR analysis enabled screening of calluses to identify somatic hybrid tissue prior to plant regeneration. The somatic hybrids were highly polyploid, mostly hexaploid (2n=6x=72), possibly due to fusion of endopolyploid protoplasts and/or chromosomal doubling during plant regeneration.     

Network analysis of oncogenic Ras activation in cancer

To investigate the unregulated Ras activation associated with cancer, we developed and validated a mathematical model of Ras signaling. The model-based predictions and associated experiments help explain why only one of two classes of activating Ras point mutations with in vitro transformation potential is commonly found in cancers. Model-based analysis of these mutants uncovered a systems-level process that contributes to total Ras activation in cells. This predicted behavior was supported by experimental observations. We also used the model to identify a strategy in which a drug could cause stronger inhibition on the cancerous Ras network than on the wild-type network. This system-level analysis of the oncogenic Ras network provides new insights and potential therapeutic strategies.     

SILICON ACCUMULATION AND ITS INFLUENCE ON SOME OF THE LEAF CHARACTERISTICS,MEMBRANE STABILITY AND YIELD IN RICE HYBRIDS AND VARIETIES GROWN UNDER AEROBIC CONDITIONS

Efforts to minimize water use in rice cultivation and stress tolerance are important in the present climate change scenario. Silica solubilizers might help in understanding the tolerance of plants to water deficit conditions or aerobic conditions. Rice cultivation in combination with silica was applied in the form of fertilizers (sodium silicate) and solubilizers (Imidazole and glycine) was studied in experimental farm and also in farmer's field. The varieties used were ‘KrishnaHamsa' (KH), ‘Rasi', ‘Jaya', hybrids ‘PA-6201' and ‘PHB-71' under aerobic conditions both in wet (Kharif) and dry (rabi) seasons. Transmission electron microscopy in this study provided evidence that silicon was deposited in the epidermal cell wall and the intercellular space of the silicon-treated rice leaves. The epidermal cell wall accumulation was absent in the control plants. Genotypic variation and treatment influences were observed for relative water content and cell membrane stability. Among the different rice cultivars ‘Rasi' followed by ‘PHB-71' and PA- 6201, were able to maintain cell membrane stability and chlorophyll content. Leaf rolling, chlorophyll, relative water contents, and dark adapted chlorophyll fluorescence were superior under aerobic conditions with application of solubilizers. However, significant differences in stomatal conductance were seen between seasons and genotypes. The silicon treated plants were able to maintain similar yields under aerobic conditions also as that of irrigated controls.     

INFLUENCE OF BORON ON SPIKELET FERTILITY UNDER VARIED SOIL CONDITIONS IN RICE GENOTYPES

Rice, grown on a wide range of soils with varying soil pH levels boron (B) availability, uptake and mobilization could be limiting hence may lead to lower productivity and rice yields has been tested at 11 locations on low yielding rice genotypes from, Initial Evaluation Trials (IET) of AICRIP viz.IET 20979, IET 21003, IET 21007, IET 21014, IET 21025 and Rasi as check variety. Active boron supplemented as foliar spray at anthesis stage at 0.2, 0.4 and 0.8 ppm. Grains with 1.20 (High density), and 1.06 (normal density) specific gravity (sp. gr.) and unfilled grains were determined. The results showed that application of boron resulted in increase in grain number (25–45) and reduced the number of unfilled spikelets. At majority of the locations, application of 0.4 ppm boron had resulted in significant increase in grain yield (4–8%). Cultures, IET 20979, IET 21007, and IET 21014 showed a positive response with the 0.4 ppm application at the majority of locations.     

Biochemistry of TBT-Degrading Marine Pseudomonads Isolated from Indian Coastal Waters

Tributyltin (TBT) is a very effective biocide and an active ingredient in antifouling paints. Screening along the Indian coast yielded 49 bacterial isolates capable of TBT assimilation. The screening was done based on the ability of bacteria to grow in mineral salt medium (MSM) containing TBT as the sole source of carbon. All the isolates produced exopolysaccharides (biosurfactants) in the medium which aid in emulsification and thus ease bioavailability of TBT. Five isolates were identified as potent TBT degraders (namely, Pseudomonas pseudoalcaligenes, Pseudomonas stutzeri, Pseudomonas mendocina, Pseudomonas putida, and Pseudomonas balearica) based on their biomass production in MSM containing TBT as the sole source of carbon. In addition to evaluating the potential of individual bacterial strains, the study also focused on using a consortium of bacteria to explore their synergistic effect when grown on TBT. Further tests like growth profile, rhamnolipid secretion profile, extracellular protein secretion profile, and detection of siderophores were performed on these isolates when grown in MSM supplemented with 2 mM TBT concentration. Emulsification activity of the crude extracellular polysaccharides against kerosene was evaluated. It can be therefore inferred that TBT degradation by these marine pseudomonads is a two-step process: (a) dispersion of TBT in the aqueous phase and (b) tin–carbon bond cleavage by siderophores affecting debutylation of TBT. The consortium of bacteria may be effective in the treatment of TBT-contaminated waste water in dry docks.     

Effect of stocking density on soil,water quality and nitrogen budget in Penaeus monodon (Fabricius, 1798) culture under zero water exchange system

Soil and water interactions and their influence on growth and production in different densities (SD8 & SD16) under zero water exchange were studied in two successive crops of Penaeus monodon in Tamil Nadu, India during 2007 and 2008. Scraping and tilling during pond preparation increased the mineralization rate than scraping only. During crop, there was no significant difference in soil organic carbon and total nitrogen between the SDs. However, as the crop progressed, organic carbon and total nitrogen content of the soil showed significant difference under both the SDs. Between the SDs, nitrate and phosphate content in water significantly differed, whereas the progress of the crop significantly increased both available and total nutrients. Mass balance of nitrogen indicated that applied feed contributed to 97.4–98.5% of input nitrogen, of which nitrogen in sediment accounted for 16.5–27.3%, nitrogen recovery in shrimp was 34.2–43.6% and the nitrogen lost through denitrification and volatilization varied from 4.7% to 34.7%. Zero water exchange system is highly efficient as nitrogen recovery is higher in shrimp and lower in discharge water. Lack of significant difference in metabolites between the SDs indicates the role of aeration and probiotics in sustaining SD16 cultures.