Sensitivity of Tilapia (Oreochromis mossambicus) to Clostridium botulinum toxins

The sensitivity of Oreochromis mossambicus to Clostridium botulinum toxin types A–E was investigated. All five toxin types were toxic to O. mossambicus. In the case of toxin types A–D, O. mossambicus was considerably more resistant than mice and, in the case of type E toxin, fish were more sensitive. The minimum intraperitoneal (i.p.) dose of type E toxin for fish was half the minimum lethal dose for mice. The results of the study suggest that good hygiene should be maintained in fish/shrimp farms to keep contamination at a low level.     

Histopathological changes in giant freshwater prawn Macrobrachium rosenbergii (de Man 1879) fed with probiotic Bacillus licheniformis upon challenge with Vibrio alginolyticus

The effect of dietary supplementation of probiotic bacterium Bacillus licheniformis on the histopathological changes in Macrobrachium rosenbergii juveniles (4.0 ± 0.02 g) challenged with known pathogenic strain of Vibrio alginolyticus are reported. Two isocaloric basal diets supplemented with probiotic bacteria B. licheniformis (1.0 × 10⁹ cfu/g feed) and other without probiotic supplementation were fed to the M. rosenbergii juveniles for 45 days. The histological observations revealed no significant changes in the hepatopancreas and gut tissues of both the experimental and the control groups which indicate that the present bacterium is a safe candidate probiont for the host. Prawns were challenged with V. alginolyticus after 45 days of feeding with probiotic diet. The histopathological studies of the hepatopancreas revealed that M. rosenbergii fed with probiotic‐supplemented diet showed less changes as compared to the prawns fed with control diet on second and fourth day of post‐experimental challenge with V. alginolyticus. Histopathological observations revealed that the gills of the prawns fed with control diet were severely affected in comparison to the prawns fed with probiotic‐supplemented diet after challenging with V. alginolyticus. Results from this study revealed the improved protection by dietary incorporation of B. licheniformis in reducing the histopathological manifestations due to V. alginolyticus infection in freshwater prawn.     

Determination of theanine in commercial tea by liquid chromatography with fluorescence and diode array ultraviolet detection

Two liquid chromatographic methods that involve precolumn derivatization with o-phthaladehyde (OPA) and phenylisothiocyanate (PITC) with fluorescence and diode array UV detection for the determination of theanine have been developed. The chromatographic separations were achieved by reverse-phase high-performance liquid chromatography using octadecyl columns and gradient elution. The methods were applied to evaluate the theanine content of commercial tea leaves. The coefficient of variation of the peak area repeatability for within day (n = 8) and between day (n = 8 over 10 days) was lower than 3% for both of the methods. The estimated limit of detection (LOD) and limit of quantitation (LOQ) for the OPA method was 0.12 and 0.35 microg theanine, respectively. The PITC method was 500-fold more sensitive with LOD and LOQ values of 0.25 and 0.75 ng, respectively. The theanine content of the commercial tea samples varied from 2-5 mg/g leaf. The overall % recoveries for these methods ranged from 93-99.3. The sensitivity and simplicity of the method render them suitable for use in quality control laboratories.     

Enhancement of beta-glucosidase and beta-galactosidase of Trigonella foenum-graecum by exposure to the allelochemical mimosine

Glycohydrolases assume significance in the metabolism of biological systems and have important industrial applications in the areas of pharmaceuticals, food, and medicine. Glycosidases were screened in germinating seeds, and attempts were made to enhance their levels. Screening of glycosidases in the seedlings during a 72 h germination period revealed higher levels of beta-glucosidase and beta-galactosidase in Trigonella foenum-graecum compared to Cicer arietinum and Vigna radiata. Activity of beta-galactosidase was in general higher than that of beta-glucosidase in all the seedlings tested. During growth, exposure of the seedlings to an allelochemical, mimosine, at 0.1 mM resulted in the enhancement of enzyme levels by 50% in the seedlings of T. foenum-graecum, whereas the addition of mimosine to the assay medium in vitro did not affect the enzyme activities. Hydrolytic activity was enhanced by addition of glycerol in the medium up to 0.1 M in the case of beta-glucosidase and with 0.05 M in the case of beta-galactosidase. In general, the hydrolytic rate was higher by about 30% in the seedlings exposed to mimosine compared to that of the control. Concomitant enhancement in the rates of transgalactosidation by 51% and transglucosidation by 23% was also noted, underscoring the relevance of plant glycohydrolases for appropriate applications.     

Development and evaluation of flow through technique for diagnosis of cystic echinococcosis in sheep

A novel in vitro flow through technique was developed and evaluated for immunodiagnosis of cystic echinococcosis in sheep using hydatid specific non-cross reactive 8-kDa protein. The 8-kDa protein was prepared from hydatid cyst fluid by DEAE-Sepharose fast flow anion exchange chromatography. In this flow through technique, the 8-kDa antigen was coated on the nitrocellulose membrane of flow through device. Protein A colloidal gold was used as detector. The evaluation of the technique was performed by comparing 150 known positive hydatid serum and known negative serum collected from sheep. The test was shown to be high sensitivity and specificity that were closely correlated with those of EITB. Furthermore the immunofiltration-based assay is rapid (2 min) and easy to perform with no requirement of special skill, reagent and instrumentation. This suggests the flow through technique is an acceptance alternative to be used in clinical laboratories lacking specialized equipments as well as large scale screening of cystic echinococcosis both in the field with animal and human populations.     

Making an Indian traditional rice variety Mahsuri, bacterial blight resistant using marker-assisted selection

Mahsuri a popular traditional variety and the first rain-fed mega variety of the Indian sub- continent. It is highly susceptible to bacterial leaf blight caused by Xanthomonas oryzae pv.oryzae. Nine best performing families of Mahsuri pyramid containing four bacterial blight resistance genes (Xa4, xa5, xa13 and Xa21) were evaluated for agronomic, yield and its related characters viz. days to 50% flowering, plant height, number of tillers, panicle length, filled grains per panicle, grain weight, and yield under natural and disease pressure conditions for three consecutive wet seasons. In addition these pyramids were also evaluated for three different spacings to find out the optimum spacing under disease free and disease pressure conditions. Results revealed that under disease free conditions there was no significant difference between the pyramids and the parent for the characters evaluated in each spacing. However characters plant height, number of tillers, panicle length, filled grains per panicle, yield per plant and yield per sq. m. showed significant variation between the different spacings across seasons. Under heavy disease pressure the parent exhibited highly susceptible reaction whereas the pyramid families were highly resistant. A wider spacing had less yield loss when compared to dense planting under BB infestation in case of parent. There was no such yield loss in the pyramid families. When yield per sq. m. was taken into consideration the 20 × 20 cm spacing showed the highest yield when compared to the other two spacings since number of plants were more. The pyramids insulated the yield loss against bacterial leaf blight and are a gain to the farmers to help overcome the heavy yield losses due to this disease. These pyramids have the potential to replace the parent and can be used directly. In addition they can be used as donors for bacterial blight resistance in any breeding program.     

Angiotensin I-converting enzyme inhibitory peptide derived from glycinin, the 11S globulin of soybean (Glycine max)

Angiotensin I-converting enzyme (ACE), a dipeptidyl carboxypeptidase, catalyzes the conversion of Angiotensin I to the potent vasoconstrictor Angiotensin II and plays an important physiological role in regulating blood pressure. Inhibitors of angiotensin 1-converting enzyme derived from food proteins are utilized for pharmaceuticals and physiologically functional foods. ACE inhibitory properties of different enzymatic hydrolysates of glycinin, the major storage protein of soybean, have been demonstrated. The IC50 value for the different enzyme digests ranges from 4.5 to 35 microg of N2. The Protease P hydrolysate contained the most potent suite of ACE inhibitory peptides. The ACE inhibitory activity of the Protease P hydrolysate after fractionation by RP-HPLC and ion-pair chromatography was ascribed to a single peptide. The peptide was homogeneous as evidenced by MALDI-TOF and identified to be a pentapeptide. The sequence was Val-Leu-Ile-Val-Pro. This peptide was synthesized using solid-phase FMOC chemistry. The IC50 for ACE inhibition was 1.69 +/- 0.17 microM. The synthetic peptide was a potent competitive inhibitor of ACE with a Ki of 4.5 +/- 0.25 x 10(-6) M. This peptide was resistant to digestion by proteases of the gastrointestinal tract. The antihypertensive property of this peptide derived from glycinin might find importance in the development of therapeutic functional foods.     

Biophysical and socio‐economic causes for increasing fallow land in Tamil Nadu

The question of increasing fallow land assumes significance at national and international levels due to the decline in net cultivated area and increasing population. Biophysical characteristics of farmland and socio‐economic conditions of farmers are responsible for the increasing fallow. This study was carried out to assess the extent of permanent fallow and identify causes for its increase. High‐resolution remote sensing data were used to identify fallow land in two identified blocks of south India, viz. Nanguneri and Kangeyam. The land resources were characterized, and the specific biophysical and socio‐economic factors responsible for the increasing fallow land were assessed. Multiple regression analysis was carried out to identify the factors responsible for increasing fallows. Soil parameters such as erosion, soil texture, soil depth and available nitrogen contributed significantly (P > 0.05) to the increase in fallow land. The size of farm holding, number of farm fragments, distance from the road in Nanguneri and farm size and the distance to the nearest town in Kangeyam were socio‐economic factors that had a strong effect (P > 0.01) on increasing the fallow land. Variable importance rankings in random forest analysis indicated that soil erosion and soil pH had the largest impact on the decision to fallow land in Nanguneri and Kangeyam blocks, respectively. Based on biophysical and socio‐economic constraints, the land resources of fallow land were evaluated for other alternate land use options and policy measures were suggested.     

Comparison of chemical characteristics of three soybean cysteine proteinase inhibitors

Three recombinant soybean cysteine proteinase inhibitors (rSCPIs) L1, R1, and N2 were chemically characterized. These inhibitors have the potential to inhibit the growth and development of three major agricultural crop pests known to utilize cysteine proteinases (CPs) for protein digestion: Western corn rootworm, Colorado potato beetle, and cowpea weevil. Characterization data obtained show differences between the inhibitors and will be needed to consider the use of rSCPIs to create insect resistance in plants.     

Bacterial microflora associated with farmed freshwater prawn Macrobrachium rosenbergii (de Man) and the aquaculture environment

The initial counts of bacteria associated with farmed giant freshwater prawn (Macrobrachium rosenbergii de Man) as well as with the water and sediment from two farms located at Kottayam district in Kerala (India) were determined. A total of 367 randomly selected isolates were characterized and identified. Prawn samples yielded mean microbiological counts of 4.92 log CFU g^?1 of shell with muscle and 7.78 log CFU g^?1 of intestine at 30°C. Bacterial numbers in the intestine of prawn were much higher than those in the pond water. Motile and non‐motile aerobic Gram‐negative bacteria together with Enterobacteriaceae accounted for 60–70% of the mesophiles isolated from water and prawn. However, the microorganisms isolated in significant numbers from shell with muscle and intestine of prawn were not recovered from the growing waters. Among Gram‐positive bacteria, Micrococcus, Bacillus, coryneforms, and Arthrobacter were found. Faecal coliforms and enterococci were found in significant numbers in M. rosenbergii. The rearing practices such as feeding and pond fertilization could have influenced the microflora in prawn. The commensal microflora of freshwater prawn included opportunistic pathogens such as Aeromonas spp. and Streptococcus spp. Storage of prawn at elevated temperatures can permit their survival and growth leading to quality loss. Care should be exercised during handling and processing to prevent contamination of edible meat.