A quantitative real‐time PCR assay for detection of Colletotrichum lindemuthianum in navy bean seeds

Bean anthracnose is a seedborne disease of common bean (Phaseolus vulgaris) caused by the fungal pathogen Colletotrichum lindemuthianum. Using seed that did not test positive for the pathogen has been proven to be an effective strategy for bean anthracnose control. To quantify the extent of anthracnose seed infection, a real‐time PCR‐based diagnostic assay was developed for detecting C. lindemuthianum in seeds of the commercial bean class navy bean. The ribosomal DNA (rDNA) region consisting of part of the18S rDNA, 5^.8S rDNA, internal transcribed spacers (ITS) 1, 2 and part of the 28S rDNA of seven races of C. lindemuthianum, 21 isolates of Colletotrichum species and nine other bean pathogens were sequenced with the universal primer set ITS5/ITS4. Based on the aligned sequence matrix, one primer set and a probe were designed for a SYBR Green dye assay and a TaqMan MGB (minor groove binder) assay. The primer set was demonstrated to be specific for C. lindemuthianum and showed a high sensitivity for the target pathogen. The detection limit of both assays was 5 fg of C. lindemuthianum genomic DNA. To explore the correlation between the lesion area and the DNA amount of C. lindemuthianum in bean seed, seeds of the navy bean cultivar Navigator with lesions of different sizes, as well as symptomless seeds, were used in both real‐time PCR assays.     

Changes in carbohydrate and nitrogenous components and amylase activities during germination of grain amaranth

Grain amaranth (Amaranthus hypochondriacus), Yercaud local variety, was soaked overnight and germinated for 192 h taking the soaked grains as the zero time (0 h) sample. The changes in the activities of - and -amylases, starch, sugar, protein and lysine contents during germination are reported. Activity of -amylase was high in the 0 h soaked grains, while -amylase activity was high in 72 h germinated grains. The joint action of the amylases resulted in a decrease of starch content from 0 to 192 h in germinated grains and an increase in total sugars during the initial period of germination.Protein nitrogen was found to decrease from 48 h to 192 h accompanied by an increase in free amino acid and non-protein nitrogen contents. Total lysine content was found to be increased by 31% in 24 h germinated grain amaranth. Protein fractionation of raw, soaked and 24 h germinated grain amaranth showed that the distribution of different types of proteins varied during germination of the grains. An increase of water soluble protein content was noticed in 24 h germinated grains.     

Effects of taurine on glutathione peroxidase, glutathione reductase and reduced glutathione levels in rats

The aim of the present study is to investigate the effects of oral administration of taurine on endogenous glutathione peroxidase (GPx) and Glutathione Reductase (GR) activities and reduced glutathione (GSH) level in normal rats. Normal saline (Group I) or 5% taurine in normal saline was administered in dose of 50 mg (Group II), 250 mg (Group III) or 500 mg kg(-1) of body weight (Group IV) through intragastric intubation for 60 days. GPx and GR enzyme activities and GSH and taurine levels were determined in liver, heart, stomach, kidney and plasma of normal Wistar rats. GPx activity showed an increase in liver, heart, stomach and plasma. GR activity increased in kidney and decreased in liver and plasma. GSH levels increased in liver, stomach and decreased in kidney. Liver showed an increase and heart, stomach and kidney a decrease in taurine level in taurine administered rats when compared to control rats. The results varied from organ to organ and the observed variations among organs might be related to their respective enzymatic, non-enzymatic antioxidant potential and its functions. From the present study it may be concluded that long term oral administration of taurine affects GPx, GR and GSH levels in normal rats.     

Genogroup I picobirnavirus in diarrhoeic foals: Can the horse serve as a natural reservoir for human infection?

ABSTRACT: Picobirnaviruses (PBV) are small, non-enveloped viruses with a bisegmented double-stranded RNA genome. In this study a PBV strain, PBV/Horse/India/BG-Eq-3/2010, was identified in the faeces of a 10 month old weaned female foal with diarrhoea in January 2010 from Kolkata, India. Surprisingly, sequence comparison and phylogenetic analysis of a short stretch of the RNA dependent RNA polymerase gene revealed close genetic relatedness (> 98% nucleotide identity) to a human genogroup I PBV strain (Hu/GPBV1) detected earlier from the same part of India. Our observations together with earlier findings on genetic relatedness between human and animal PBV warrant further studies on zoonotic potential.     

Phenolics and Antioxidative Activities in Narrow-Leafed Lupins (Lupinus angustifolius L.)

Eight lupin (Lupinus angustifolius L.) genotypes grown at four locations in south central Alberta in 2004 were evaluated for variability in phenolic constituents and antioxidant activity measured by a photochemiluminescence assay. Genotype was the main source of variation for content of phenolic compounds and antioxidant activities. Phenolic compounds in genotypes varied minimally from 11.9 to 14.7 mg catechin equivalent and 4.15 to 4.95 mg rutin equivalent g⁻¹ lupin for total phenolic and flavonoid contents, respectively. Lupin genotypes exhibited weak antioxidant activity based on water–soluble substances (ACW) of 0.54 to 1.07 μmole Trolox equivalent antioxidant capacities (TEAC)/g with lag time ranging from 70 to 153 s and an antioxidant index of 6.7 to 14.5 and 1.9 to 3.3 μmole TEAC/g based on measurements of lipid-soluble substances (ACL). Antioxidant activity of lupin genotypes was not related to phenolic contents of seeds.     

Effect of fenvalerate on oxidative stress biomarkers in the brackish water prawn Penaeus monodon

The toxicity of fenvalerate to the prawn Penaeus monodon was evaluated using biomarkers of stress. In a preliminary bioassay test, P. monodon was exposed to a series of fenvalerate concentrations, which showed 4, 6.5 and 8.5 μg L⁻¹ to be sublethal, median lethal and lethal, respectively. Sublethal effect of fenvalerate was further evaluated in hepatopancreas, muscle and gills of prawns with reference to oxidative stress biomarkers. Significant induction of lipid peroxidation and glutathione-S-transferase activity was found in hepatopancreas, muscle and gills of prawns exposed to fenvalerate when compared to control (P < 0.001, P < 0.05 and P < 0.05). On the contrary, the activities of Superoxide dismutase, catalase, glutathione peroxidase, vitamin C, vitamin E and glutathione were found to be reduced in the experimental group of prawns when compared to control. The results suggest that the animals were under oxidative stress when exposed to sublethal concentration of fenvalerate.     

Effect of Naphthalene on Phosphatases and Esterase in Muscle and Ovary of Intermoult Marine Edible Female Crab,Scylla Serrata

The effects of naphthalene on glycogen, lipid, phosphatases and esterase have been studied in the muscle and ovary of Intermoult marine female crab, Scylla serrata. Rearing the crab in increasing sublethal concentrations of naphthalene for 96 hr (i.e. 0.005, 0.010, 0.015 and 0.020 mg L-1). The results show an overall decrease in the glycogen, lipid, alkaline phosphatase and esterase in muscle and ovary. On the other hand, acid phosphatase activity in the muscle and ovary has increased.     

Effect of buprofezin on adult life span,oviposition, egg hatch and progeny production of the cotton whitefly,Bemisia Tabaci

The 50% survival ofBemisia tabaci Genn. females was reduced to 5.9 days by exposure to bean seedlings treated with 1000 ppm buprofezinvs 8.9 days in the control. At lower concentrations ranging from 50 to 500 ppm, survival was extended to 11.8–14.2 days. Mean fecundity was significantly reduced on plants treated with 1000 ppm as compared with the control, whereas at 50 to 500 ppm the oviposition period was extended with a considerable increase in mean fecundity, irrespective of concentrations. Overall mean percent hatch dropped gradually from 25.8 to 12.9 with increase of concentration from 50 to 1000 ppm, as against 96.9% in the control. The survival of hatched nymphs was greatly affected at all buprofezin concentrations. No immature stages survived at 50, 100, 250 and 500 ppm; nymphal survival was 13.8, 7.0, 1.9 and 1.2%, respectively, as against 81.6% in the control.     

A rare case of globosus amorphus in a goat

We present a case of globosus amorphus delivered from a goat and subjected to radiography and histological examination. Radiography revealed a lack of development of any organ system; histological sections showed evidence of lymphoid aggregations, mononuclear infiltrations, blood capillaries, and dense fibroblasts.     

Cellular Composition and Viability of Cloned Bovine Embryos Using Exogene-Transfected Somatic Cells

The present study compared the efficiency of transgenic (TG) cloned embryo production by somatic cell nuclear transfer (SCNT) with fetal-derived fibroblast cells (FFCs) which were transfected with pEGFP-N1 to in vitro-fertilized (IVF), parthenogenetic and SCNT counterparts by evaluating the rates of cleavage and blastocyst formation, apoptosis rate at different developmental stages, cell number, ploidy and gene expression in blastocysts. In SCNT and TG embryos, the rates of cleavage and blastocyst formation were significantly lower (p < 0.05) than those of IVF controls, but it did not differ between SCNT and TG embryos. In IVF control, 86.7% embryos displayed diploid chromosomal complements and the rates were significantly (p < 0.05) higher than those of SCNT and TG embryos. Most TG embryos (79%) with FFCs expressed the gene by both PCR and under fluorescence microscopy. The expression of apoptosis by TUNEL was first detected at six to eight cell stages in all embryos of IVF, SCNT and TG groups, but the expression rate at each developmental stages was significantly higher (p < 0.05) in SCNT and TG embryos than in IVF counterparts. The expression rate in inner cell mass (ICM) of TG embryos was significantly higher (p < 0.05) than in SCNT and IVF embryos. These results indicate that the high occurrence of apoptosis observed in SCNT and TG embryos compared with IVF counterparts might influence the developmental competence. Moreover, the SCNT embryos derived using non-transfected donor cells exhibited a lower apoptosis expression in ICM cells than in TG embryos derived using pEGP-N1-transfected donor cells suggesting a possible role of negative gene effect in TG embryos.