The qualitative assessment of glycosaminoglycans in the canine intervertebral disc using a critical electrolyte concentration staining technique.

Through the use of a critical electrolyte concentration staining technique, the glycosaminoglycans (GAGs) in the nucleus pulposus region of the canine intervertebral disc were arbitrarily identified as "hyaluronic acid", chondroitin sulphate and keratan sulphate. Approximate estimates of GAG concentration could be qualitatively deduced by determining the appropriate GAG "ALCIANOPHILIC INDEX". This index was considered to be an effective qualitative adjunct to chemical quantitation of GAGs in the intervertebral disc.     

Carbon sequestration and relationship between carbon addition and storage under rainfed soybean–wheat rotation in a sandy loam soil of the Indian Himalayas

Soil organic matter (SOM) contributes to the productivity and physical properties of soils. Although crop productivity is sustained mainly through the application of organic manure in the Indian Himalayas, no information is available on the effects of long-term manure addition along with mineral fertilizers on C sequestration and the contribution of total C input towards soil organic C (SOC) storage. We analyzed results of a long-term experiment, initiated in 1973 on a sandy loam soil under rainfed conditions to determine the influence of different combinations of NPK fertilizer and fertilizer + farmyard manure (FYM) at 10 Mg ha⁻¹ on SOC content and its changes in the 0–45 cm soil depth. Concentration of SOC increased 40 and 70% in the NPK + FYM-treated plots as compared to NPK (43.1 Mg C ha⁻¹) and unfertilized control plots (35.5 Mg C ha⁻¹), respectively. Average annual contribution of C input from soybean (Glycine max (L.) Merr.) was 29% and that from wheat (Triticum aestivum L. Emend. Flori and Paol) was 24% of the harvestable above-ground biomass yield. Annual gross C input and annual rate of total SOC enrichment were 4852 and 900 kg C ha⁻¹, respectively, for the plots under NPK + FYM. It was estimated that 19% of the gross C input contributed towards the increase in SOC content. C loss from native SOM during 30 years averaged 61 kg C ha⁻¹ yr⁻¹. The estimated quantity of biomass C required to maintain equilibrium SOM content was 321 kg ha⁻¹ yr⁻¹. The total annual C input by the soybean–wheat rotation in the plots under unfertilized control was 890 kg ha⁻¹ yr⁻¹. Thus, increase in SOC concentration under long-term (30 years) rainfed soybean–wheat cropping was due to the fact that annual C input by the system was higher than the required amount to maintaining equilibrium SOM content.     

Development of mulberry varieties for sustainable growth and leaf yield in temperate and subtropical regions of India

Sericulture is an agro-based industry, which heavily depends on mulberry (Morus spp.) for its sustainability, as mulberry leaf is the only available feed for the silkworm Bombyx mori L. Sericulture, in West Bengal and other subtropical regions in India, suffers from lack of adequate quantity of quality mulberry leaf during the colder months, the best season for rearing the high yielding bivoltine silkworm hybrids. In order to develop mulberry varieties, which can sustain normal growth during these colder months to yield better, efforts have been made in different research institutes in India. The present report is part of such an effort in which accessions in germplasm bank were screened for better growth, leaf retention and leaf yield contributing associated traits. Twenty three selected parents were crossed, 3,500 seeds were sown, 2,700 seedlings were transplanted to progeny row trail, selected 210 hybrids were evaluated under primary yield trial and finally nine hybrids along with a control variety were studied in detail for all leaf yield contributing traits under final yield trial. Stability analysis was adopted to identify hybrids, which can yield stably across seasons. The hybrids CT-44 and CT-11 out yielding the control by 17.17 and 7.11% were selected as these hybrids yield 7.93 and 8.15 mt/ha leaf respectively during the colder months (February) for their direct use as cultivars in West Bengal and other subtropical areas of India to sustain bivoltine sericulture to produce quality and gradable silk fibers.     

Development of transgenic fungi that kill human malaria parasites in mosquitoes

Metarhizium anisopliae infects mosquitoes through the cuticle and proliferates in the hemolymph. To allow M. anisopliae to combat malaria in mosquitoes with advanced malaria infections, we produced recombinant strains expressing molecules that target sporozoites as they travel through the hemolymph to the salivary glands. Eleven days after a Plasmodium-infected blood meal, mosquitoes were treated with M. anisopliae expressing salivary gland and midgut peptide 1 (SM1), which blocks attachment of sporozoites to salivary glands; a single-chain antibody that agglutinates sporozoites; or scorpine, which is an antimicrobial toxin. These reduced sporozoite counts by 71%, 85%, and 90%, respectively. M. anisopliae expressing scorpine and an [SM1](8):scorpine fusion protein reduced sporozoite counts by 98%, suggesting that Metarhizium-mediated inhibition of Plasmodium development could be a powerful weapon for combating malaria.     

Evaluation of the nutritive value of Leucaena leucocephala leaf meal, inoculated with fish intestinal bacteria Bacillus subtilis and Bacillus circulans in formulated diets for rohu, Labeo rohita (Hamilton) fingerlings

Nine isonitrogenous (35% crude protein approximately) and isocaloric (18.37 kJ g^?1) experimental diets (RLL20–BCFL40) were formulated with either raw or treated (inoculated with fish intestinal bacteria) Leucaena leucocephala leaf meal at 20%, 30% and 40% levels replacing other ingredients partially from a fish meal based reference diet (RD). Two specific strains of fish intestinal bacteria, Bacillus subtilis (isolated from Cyprinus carpio) and B. circulans (isolated from Oreochromis mossambicus) having extracellular cellulolytic and amylolytic activities, were used to inoculate Leucaena leaf meal for 15 days at 37°C. The crude fibre, cellulose and hemicellulose contents and the antinutritional factors, tannin, phytic acid and mimosine in the leaf meal decreased due to inoculation. However, free amino acids and fatty acids increased in the treated leaf meal. The response of rohu, Labeo rohita, fingerlings fed the experimental diets for 80 days was compared with fish fed a RD. Both the inclusion level and type of Leucaena leaf meal in diets significantly affected the growth performance of rohu. Fish fed diets containing inoculated Leucaena leaf meal performed better in comparison with those with the RD. On the basis of growth response, feed conversion ratio, protein efficiency ratio and apparent net protein utilization, diet formulated with 30%Leucaena leaf meal inoculated with B. circulans resulted in the best performance of rohu fingerlings followed by diet with 40%B. subtilis inoculated Leucaena leaf meal. The apparent protein digestibility (APD) was better in fish fed diets containing B. circulans inoculated leaf meal. An increasing level of raw Leucaena leaf meal was associated with a decrease in the carcass protein content of rohu fingerlings. The activity of α‐amylase increased with the increasing level of treated leaf meal in diets. Cellulase activity increased with increasing level of inclusion of raw leaf meal, and was comparatively lower in fish fed diets with treated leaf meal. Activities of protease and lipase were higher in fish fed the RD. The results showed that it is possible to incorporate Leucaena leaf meal inoculated with enzyme‐producing fish intestinal bacteria in carp diets up to 40% level of inclusion.     

Determination of inter- and intra-species genetic relationships among six Eucalyptus species based on inter-simple sequence repeats (ISSR)

Eucalyptus is the most economically important hardwood plantation tree cultivated in tropical and subtropical countries. Inter-simple sequence repeat (ISSR) markers were used to evaluate genetic relationships within and between individuals of six Eucalyptus species. A total of 583 loci (265 to 1535 bp) were amplified from 149 individuals belonging to the six Eucalyptus species using seven ISSR primers (two to three nucleotide repeats anchored with one or two nucleotides at the 3' or 5' region). The ISSR fragments indicated significant polymorphism and genetic diversity among the individuals. Cluster analysis and principal component analysis revealed the occurrence of wide genetic diversity among populations of E. tereticornis Sm., E. camaldulensis Dehnh. and E. urophylla S.T. Blake and narrow genetic diversity among populations of E. citriodora Hook. and E. grandis W. Hill ex Maiden. Genetic diversity was high in E. tereticornis Sm. (47.27%) and low in E. citriodora (18.64%). Maximum Nei's genetic identity (0.897) was observed between E. camaldulensis and E. tereticornis species, whereas maximum genetic diversity (0.286) was found between individuals of E. citriodora and E. grandis.     

Prevalence,molecular fingerprinting and drug resistance profile of enterovirulent <Emphasis Type="Italic">Escherichia coli</Emphasis> isolates from free-ranging yaks of Tawang district,Arunachal Pradesh,India

Of 273 samples (rectal swab) collected from free-ranging yaks of Tawang district, Arunachal Pradesh, 42 Shiga toxin-producing Escherichia coli (STEC), six enteropathogenic E. coli (EPEC) and 27 enterotoxigenic E. coli (ETEC) strains were isolated. All the STEC and EPEC strains were further investigated for respective stx variants (for STEC only) and additional putative virulence factors. The 27 ETEC strains were also screened for characteristic enterotoxin gene(s) and colonization factors. Occurrence of ETEC was significantly (p < 0.05) higher in the diarrheic yaks and yaks of less than 1 year of age. Majority of enterovirulent E. coli isolates were resistant to amikacin, azithromycin, chloramphenicol, colistin, doxycycline, furazolidone, nalidixic acid, nitrofurantoin, streptomycin and tetracycline. Dendrogram, constructed with molecular fingerprinting profiles obtained from RAPD (Randomly Amplified Polymorphic DNA) and ERIC (Enterobacterial Repetitive Intergenic Consensus) PCR, placed the isolates in different clusters irrespective of their serotypes, virulence gene and drug resistance pattern. Collectively, the study indicates that yaks, being a potential reservoir of multidrug resistant STEC and EPEC, may represent significant risk to public health in this region. Higher recovery of ETEC isolates from yaks with diarrhea points out that ETEC may be a major determinant for repeated occurrence of diarrhea in yaks.     

Phytic Acid‐Induced Inhibition of Digestive Protease and α‐Amylase in Three Indian Major Carps: An in vitro Study

Antinutritional effects of phytic acid (myoinositol hexaphosphate, IP6) on growth and digestibility in fish have been reported. However, specific effect of IP6 on the digestive enzymes in fish has not been addressed. In this study, inhibitory effect of synthetic IP6 (Phytic acid sodium salt, 90% purity) on the activity of the digestive protease and α‐amylase in rohu, Labeo rohita; catla, Catla catla; and mrigal, Cirrhinus mrigala has been investigated in vitro. Graded levels (12.5, 25, 50, 100, 150, and 200 µg/mL) of IP6 were added to the reaction mixtures containing enzyme extracts and substrate solution in triplicate to detect any change in enzyme activity. Results of the experiment revealed that IP6 significantly inhibit/lower activities of the digestive enzymes in a dose‐dependent manner, as evident from the regression equations (F values significant at P < 0.001 level). Apparently, irrespective of the fish species studied IP6‐induced inhibition of α‐amylase activity was greater than protease activity. Among the three fish species studied, C. mrigala appeared to be more sensitive to IP6 for both α‐amylase and protease activity. Enzyme activity was least affected in C. catla. Results of the study might raise concern while incorporating IP6 rich plant‐derived feed ingredients in aqua feed preparation.     

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.     

Extension of shelf life of tuberose flowers using a combination of gamma irradiation and generally regarded as safe (GRAS) preservatives and assessment of antimicrobial potency of senesced flowers

Tuberose cut flowers, available as loose flowers, were treated with gamma (γ) irradiation and generally regarded as safe (GRAS) preservative solutions for extension of shelf life. The flowers were packaged in low-density polyethylene bags, heat sealed and stored at 23 ± 2°C, 80% relative humidity (RH) and 4 ± 1°C, 40% RH, respectively. The flowers stored at these two temperature regimes were subjected to sensory evaluation and biochemical analyses. From these assessments, the longest shelf life of tuberose flowers was found to be 8 days at 23 ± 2°C, 80% RH (compared to 4 days for control) and 24 days at 4 ± 1°C, 40% RH (compared to 8 days for control) using combination treatment of low dose γ-irradiation (0.02 kGy) and preservative solutions (4% sucrose and 0.02% CaCl₂). Ethanolic extract of tuberose flowers of the most shelf stable set (stored at 4 ± 1°C), i.e. at the end of 24 days, showed antimicrobial potency against the common skin pathogen Staphylococcus aureus (ATCC 25923 and MDR strains), suggesting utility of the senesced tuberose flowers for therapeutic applications. This preservation technique would promote export of tuberose flowers by extension of their shelf lives and allow utilization of these flowers, post senescence.