Purification and characterization of a 630 kDa bacterial killing metalloprotease (KilC) isolated from plaice Pleuronectes platessa (L.), epidermal mucus

Antibacterial chemicals in the mucus of fish such as lysozyme, lectins, peptides and proteases provide an efficient first line of defence against pathogens. This study shows that there are at least three antibacterial proteins in plaice skin mucus in addition to lysozyme. One of these proteins is responsible for approximately 74% of the antibacterial activity and is a 630 kDa protease complex designated KilC (bacterial killing metalloprotease C). Purified KilC kills the bacteria Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa efficiently. The protease activity of KilC is dependent upon the divalent cation Mg(2+) and shows pH dual optima of 5.0 and 8.0. The enzyme has a temperature optimum of 25 degrees C and is made up of at least five different sized peptides. Studies with protease inhibitors show that the catalytic site of KilC may be cysteine- or serine protease-like. KilC may kill bacterial cells by acting directly upon the bacteria or by producing low molecular weight bioactive compounds such as peptides.     

Enhancing the mineral and vitamin content of wheat and maize through plant breeding

More than half of the world's population suffers micronutrient undernourishment. The main sources of vitamins and minerals (iron, zinc, and vitamin A) for low-income rural and urban populations are staple foods of plant origin that often contain low levels or low bioavailability of these micronutrients. Biofortification aims to develop micronutrient-enhanced crop varieties through conventional plant breeding. HarvestPlus, the CGIAR's biofortification initiative, seeks to breed and disseminate crop varieties with enhanced micronutrient content that can improve the nutrition of the “hard to reach” (by fortification or supplementation programmes) rural and urban poor in targeted countries/regions. In attempting to enhance micronutrient levels in maize and wheat through conventional plant breeding, it is important to identify genetic resources with high levels of the targeted micronutrients, to consider the heritability of the targeted traits, to explore the availability of high throughput screening tools and to gain a better understanding of genotype by environment interactions. Biofortified maize and wheat varieties must have the trait combinations which encourage adoption such as high yield potential, disease resistance, and consumer acceptability. When defining breeding strategies and targeting micronutrient levels, researchers need to consider the desired micronutrient increases, food intake and retention and bioavailability as they relate to food processing, anti-nutritional factors and promoters. Finally, ex ante studies are required to quantify the burden of micronutrient deficiency and the potential of biofortification to achieve a significant improvement in human micronutrient status in the deficient target population in order to determine whether a biofortification program is cost-effective.     

Development and characterization of a cell line TTCF from endangered mahseer Tor tor (Ham.)

Tor tor is an important game and food fish of India with a distribution throughout Asia from the trans-Himalayan region to the Mekong River basin to Malaysia, Pakistan, Bangladesh and Indonesia. A new cell line named TTCF was developed from the caudal fin of T. tor for the first time. The cell line was optimally maintained at 28°C in Leibovitz-15 (L-15) medium supplemented with 20% fetal bovine serum (FBS). The propagation of TTCF cells showed a high plating efficiency of 63.00%. The cytogenetic analysis revealed a diploid count of 100 chromosomes at passage 15, 30, 45 and 60 passages. The viability of the TTCF cell line was found to be 72% after 6 months of cryopreservation in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 578- and 655-bp sequences of 16S rRNA and cytochrome oxidase subunit I (COI) genes of mitochondrial DNA (mtDNA) respectively. TTCF cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids. Further, immunocytochemistry studies confirm its fibroblastic morphology of cells. Genotoxicity assessment of H?O? in TTCF cell line revealed the utility of TTCF cell line as in vitro model for aquatic toxicological studies.     

Immunohistochemical study on changes in gill Na+/K+-ATPase α-subunit during smoltification in the wild masu salmon, Oncorhynchus masou

Changes in immunoreactivity of Na⁺/K⁺-ATPase -subunit in gill sections of wild masu salmon (Oncorhynchus masou) during the parr-smolt transformation (smoltification) were compared with changes in gill Na⁺/K⁺-ATPase specific activity. Gill Na⁺/K⁺-ATPase specific activity increased from April and peaked in May. Immunohistochemical analysis, using an antiserum against a synthetic oligopeptide based on the conserved region of the Na⁺/K⁺-ATPase -subunit, revealed that immunoreactivity was confined to chloride cells in the surface layer of primary lamellae and the proximal end of secondary lamellae. The size and number of these cells increased gradually from February to May; however, the number of chloride cells of the secondary lamellae decreased in May. These data suggest that the synthesis of Na⁺/K⁺-ATPase and the proliferation of chloride cells occur prior to the elevation of enzyme activity. Moreover, it is likely the proliferation and hypertrophy of chloride cells on primary lamellae prepare smolts for entry into seawater and migration in the ocean.     

EFFICACY OF GYPSUM AND MAGNESIUM SULFATE AS SOURCES OF SULFUR TO RAPESEED IN LATERITIC SOILS

A field experiment conducted on rapeseed (Brassica juncea L.) during 2005–2006 in a typical lateritic soil (Alfisol) of West Bengal, India revealed that sources of sulfur viz. gypsum and magnesium sulfate and levels of sulfur (0, 20, 40, 60 kg S ha^?1) have significant influence on grain yield, total biological yield, sulfur concentration in grain and stover, total sulfur uptake, oil content and oil yield and chlorophyll content. The maximum grain yield (18.28 q ha^?1) and oil yield (8.59 q ha^?1) was obtained with magnesium sulfate followed by gypsum yielded the grain yield of 17.99 q ha^?1 and oil yield of 8.22 q ha^?1 at 40 kg S ha^?1. Overall, the best performance was recorded when sulfur was applied at 40 kg S ha^?1 either as magnesium sulfate or gypsum. Results revealed that magnesium sulfate may be considered as the better source of sulfur than gypsum to raise the mustard crop in sulfur deficient acidic red and lateritic soils of West Bengal and if farmers apply either magnesium sulfate or gypsum to soils, the possible deficiency of sulfur and magnesium/calcium in soils and plants can be avoided.     

Effect of amending materials on growth of radish plant in salinized soil

The critical Na levels in soil which restricted seed germination and growth of radish, and effect of the amending materials on reducing the salinity hazard were determined by performing petri dish and pot experiments. The plants were grown in amended salinized soil in a phytotron for 21 days. Excess accumulation of Na in cells suppressed seed germination and plant growth. The critical Na level in control soil which restricted germination and growth was 12.5mg Na/g soil. At the same salinity level, amending materials promoted seed germination and plant growth mainly by inhibiting the excess flow of Na into the cell through ion holding mechanism. Among the materials used, the commercial manure of 4 months old held greater amount of Na(3.7mg/g) and thus more efficiently reduced the ion damage. Plants grown under such treatment had the highest dry matter yield (28.Gmg/plant), water content was normal(84%), and the ratio for Na in root to Na in plant's shoot was 2.4. As for the case of plant grown in control soil the respective data were 8. 5mg, 54%, and 4.8.     

Simulation Model for Composting Cellulosic (Bagasse) Substrates

A dynamic model of aerobic composting with solid substrates was developed as a first step to predict the rate of decomposition of cellulosic materials. The model was based on heat and mass transfer principles and reaction kinetics of biological processes. The relations describing the rates of reaction and the degradability potential of the substrates were determined based on laboratory-scale experiments. The model was developed using a graphical software package, Stella II® (which was able to solve feedback loops encountered in actual composting processes.

The model was validated with temperature, oxygen distribution and mass degradation accompanying the composting of bagasse. Experimental setups consisted of 30 gallon drum reactors, continuously aerated. Temperature and oxygen in exhaust gases were monitored on an hourly basis and measurements of moisture content, pH and volatile solids were done on a weekly basis.

The data fit with the model was excellent for mass degraded and temperatures for the first 10 days. From day 10 to 14, the deviations between model predictions (as solved with Stella II®) and actual measurements did not exceed 2.6 percent. The concentrations of oxygen in outlet gases showed fair agreement between predicted and experimental values. Simulating the model with varying airflow rates showed that the maximum mass breakdown rates occurred for airflow rates of 0.12 kg dry air/(kg dry compost.hr) while maintaining the temperatures below 50°C, through a temperature feedback control strategy.     

Calculating the density of loamy sandy soddy-podzolic soils from penetration resistance diagrams

The possibility for the calculation of the soil bulk density from two-dimensional penetration resistance diagrams drawn using a penetrometer during the vertical sounding of the soil has been discussed. A method has been proposed for calculating the bulk density of loamy sandy soddy-podzolic soils from the soil penetration test diagrams. The model for the calculation of the soil bulk density is based on the general physical concepts with the involvement of similarity theory methods. The adequacy and quality of the model have been studied statistically. The model has been found to be suitable for calculating the bulk density of loamy sandy and sandy podzolic soils.     

Changes in the pore space of soil constructions in physical laboratory models

The dynamics of pore space structure in different filled soil constructions during water infiltration and wetting–drying processes is studied. Model laboratory experiments in columns physically simulate water infiltration after penetration at a rate of 600 mm with free outflow from the lower end of the column followed by multiple drying of soil constructions composed by alternating layers of sand, peat, and A and В horizons of soddy-podzolic soil. In two- and three-dimensional tomographic images, changes in the pore space and the interpenetration of solid phase at the boundaries between individual horizons of soil constructions are analyzed.