Molecular characterization of Streptococcus agalactiae and Streptococcus uberis isolates from bovine milk

Streptococci are one among the major mastitis pathogens which have a considerable impact on cow health, milk quality, and productivity. The aim of the present study was to investigate the occurrence and virulence characteristics of streptococci from bovine milk and to assess the molecular epidemiology and population structure of the Indian isolates using multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Out of a total of 209 bovine composite milk samples screened from four herds (A–D), 30 Streptococcus spp. were isolated from 29 milk samples. Among the 30 isolates, species-specific PCR and partial 16S rRNA gene sequence analysis identified 17 Streptococcus agalactiae arising from herd A and 13 Streptococcus uberis comprising of 5, 7, and 1 isolates from herds B, C, and D respectively. PCR based screening for virulence genes revealed the presence of the cfb and the pavA genes in 17 and 1 S. agalactiae isolates, respectively. Similarly, in S. uberis isolates, cfu gene was present in six isolates from herd C, the pau A/skc gene in all the isolates from herds B, C, and D, whereas the sua gene was present in four isolates from herd B and the only isolate from herd D. On MLST analysis, all the S. agalactiae isolates were found to be of a novel sequence type (ST), ST-483, reported for the first time and is a single locus variant of the predicted subgroup founder ST-310, while the S. uberis isolates were found to be of three novel sequence types, namely ST-439, ST-474, and ST-475, all reported for the first time. ST-474 was a double locus variant of three different STs of global clonal complex ST-143 considered to be associated with clinical and subclinical mastitis, but ST-439 and ST-475 were singletons. Unique sequence types identified for both S. agalactiae and S. uberis were found to be herd specific. On PFGE analysis, identical or closely related restriction patterns for S. agalactiae ST-483 and S. uberis ST-439 in herds A and B respectively, but an unrelated restriction pattern for S. uberis ST-474 and ST-475 isolates from herds D and C respectively, were obtained. This signifies that the isolates of particular ST may exhibit related PFGE patterns suggesting detection of a faster molecular clock by PFGE than MLST. Since all the isolates of both the species belonged to novel sequence types, their epidemiological significance in global context could not be ascertained, however, evidence suggests that they have uniquely evolved in Indian conditions. Further research would be useful for understanding the role of these pathogens in bovine sub-clinical mastitis and implementing effective control strategies in India.     

Cold tolerance at the early growth stage in wild and cultivated rice

The present study was conducted to understand the pattern of variation and the genetic bases for cold tolerance at the early growth stage in Asian rice. The genetic variation was investigated at the germination, plumule and seedling stages among 57 strains including cultivated rice (Oryza sativa ssp. indica and ssp. japonica) and its wild progenitor (Oryza rufipogon). The significant differentiation of cold tolerance was observed among the taxonomically divided groups. At the germination stage, both indica and japonica subspecies tended to be more tolerant than O. rufipogon, whereas at the plumule and seedling stages, ssp. japonica tended to be more tolerant than ssp. indica and O. rufipogon. Furthermore, in cold tolerance at the plumule stage, the clinal variation across the latitude of origins was observed within O. rufipogon and ssp. japonica, suggesting that the current pattern of variation seems to have been shaped by both their phylogenetic histories and on-going adaptation to the local environments. QTL analysis between O. sativa ssp. japonica (tolerant) and O. rufipogon (susceptible) revealed five putative QTLs for cold tolerance at the plumule and seedling stages but not at the germination stage. Substitution mapping was also carried out to precisely locate the two major QTLs for cold tolerance at the plumule stage, which could be used for improvement of tolerance to cold stress in ssp. indica.     

Comparative evaluation of fermented and non-fermented de-oiled rice bran with or without exogenous enzymes supplementation in the diet of <Emphasis Type="Italic">Labeo rohita</Emphasis> (Hamilton, 1822)

A 60-day feeding trial was conducted to study the effect of exogenous enzymes (xylanase and phytase) supplementation in the non-fermented and fermented de-oiled rice bran (DORB)-based diet of Labeo rohita. Four test diets (T1-DORB-based diet, T2-fermented DORB-based diet, T3-phytase and xylanase supplemented DORB-based diet, and T4-phytase and xylanase supplemented fermented DORB-based diet) were formulated and fed to the respective groups. Test diets T3 and T4 were supplemented with 0.01% xylanase (16,000 U kg^?1) and 0.01% phytase (500 U kg^?1) enzymes. One hundred twenty juveniles of L. rohita, with an average weight 5.01?±?0.02 g, were stocked in 12 uniform size plastic rectangular tanks in triplicate with 10 fishes per tank following a completely randomized design (CRD). Exogenous enzyme supplementation to the T3 group significantly improved the growth performance of L. rohita (p?<?0.05). Fermented DORB fed groups registered significantly lower growth irrespective of the supplementation of exogenous enzymes. The carcass composition (except CP %), enzyme activities (except amylase activity), globulin, and A/G ratio did not vary significantly (p?>?0.05). Based on the results of the present study, it is concluded that exogenous enzyme supplementation significantly increases the growth of fish fed with DORB-based diet.     

Diversity analysis and evaluation of wild <Emphasis Type="Italic">Abelmoschus</Emphasis> species for agro-morphological traits and major biotic stresses under the north western agro-climatic condition of India

The wild species in general is considered to be the reservoir of genes especially for biotic and abiotic stresses. In okra, the predominant biotic stresses are yellow vein mosaic disease (YVMD), shoot and fruit borer and leaf hopper. Sixty eight (68) accessions belonging to four wild Abelmoschus species [Abelmoschus caillei (A. Chev.) Stevels, Abelmoschus manihot (L.) Medik., Abelmoschus moschatus (L.) Medik. and Abelmoschus tuberculatus Pal et Singh] and eight okra varieties were characterized and evaluated for phenological characters including biotic stresses under natural epiphytotic condition. The wild species examined consisted of 18 accessions (16 exotic and 2 indigenous) of A. caillei, 29 accessions of A. manihot, 16 accessions of A. moschatus and 5 accessions of A. tuberculatus. All the wild Abelmoschus species exhibited high diversity (as measured by Shannon Diversity Index) for 3 qualitative characters viz. intensity of stem colour, leaf shape, epicalyx shape, 13 quantitative characters and 3 biotic stress parameters. Among the wild species, A. caillei and A. tuberculatus showed maximum and minimum diversity for qualitative characters, respectively. There was significant variation for 19 out of 24 quantitative characters studied. Inter-species diversity pattern as estimated through Ward’s Minimum Variance Dendrogram and Principal Component Analysis revealed clear differentiation among the species with minimum overlapping indicating close association between geographical origins and clustering pattern. Intra-species diversity indicated role of specific adaptation in sub-clustering. Resistance to YVMD was found in accessions belonging to three wild species viz. A. caillei, A. manihot and A. moschatus while resistance to shoot and fruit borer and leaf hopper was found in accessions of all the four wild species. The resistant accessions can further be used for introgressing biotic stress resistance through pre-breeding into cultivated okra species.     

Evaluation of the persistent activity of moxidectin (10%) long-acting (LA) injectable formulation against Dictyocaulus viviparus, Haemonchus placei, Trichostrongylus axei and Oesophagostomum radiatum infections in cattle

Two controlled studies were conducted to evaluate the persistent efficacy of moxidectin (10%) long-acting (LA) injectable formulation against Dictyocaulus viviparus, Haemonchus placei, Trichostrongylus axei and Oesophagostomum radiatum in cattle. The moxidectin LA injectable formulation was administered as a single subcutaneous injection into the proximal third of the ear at a dose rate of 0.01 ml/kg BW to provide 1.0 mg moxidectin/kg BW. The product had persistent efficacy of >90% against D. viviparus, H. placei and Oe. radiatum for at least 150 days post-treatment and against T. axei for at least 90 days post-treatment.     

Zinc fertilization in rice-wheat cropping system under upland calcareous soil

Zinc (Zn) fertilization is important for Zn crop biofortification as well as increasing yields, thus proper Zn recommendations for soil application is needed for Zn deficient soils. The effectiveness of Zn applications was evaluated in different combinations of rates (2.5, 5.0, 7.5, and 10.0?kg?ha^?1 per year) and frequencies (initial, alternate, and every year) in rice (Oriza sativa L.) – wheat (Triticum aestivum L.) cropping system in a Zn-deficient upland calcareous soil in the fourth year. Zn applications to rice at 7.5 and 10?kg?ha^?1 of alternate year and 5.0 to 10?kg?ha^?1 of every year had the highest rice equivalent yield as compared to no-Zn treatment. Hence, Zn application to rice at 7.5?kg?ha^?1 at alternate years is the lowest rate at which highest rice equivalent yield of rice-wheat cropping system can be obtained.     

Enhanced Erythrocytic Lipid Peroxides and Reduced Plasma Ascorbic Acid,and Alteration in Blood Trace Elements Level in Dairy Cows with Mastitis

Oxidative stress has been associated in several inflammatory conditions and incriminated in the pathogenesis of many diseases. However, little information is available on the status of plasma antioxidant levels, essential components of important antioxidant enzymes such as copper, zinc and selenium in blood, and the end product of oxidative damage to the erythrocytic polyunsaturated fatty acids in inflammatory udder conditions. Blood samples were collected from three groups of dairy cows, with 21 in each group: animals with healthy udder, clinical mastitis, and subclinical mastitis. These animals were randomly selected from a herd on the basis of the California mastitis test, somatic cell count and total bacterial count. The mean plasma ascorbic acid concentration was significantly lower in cows with subclinical (p = 0.004) and clinical mastitis (p = 0.000) and the erythrocytic lipid peroxide levels were significantly (p = 0.000) higher in clinical mastitis as compared to controls. There was a significant decrease in mean blood zinc concentration in subclinical (p = 0.005) and clinical mastitis (p = 0.000), but an increase in mean blood copper level in the clinical mastitis group. It was concluded that the blood antioxidant status declines in inflammatory udder conditions, suggesting that incorporation of antioxidants may help in better management of mastitis in dairy cows.     

Therapeutic and persistent efficacy of moxidectin 1% nonaqueous injectable formulation against natural and experimentally induced lung and gastrointestinal nematodes in cattle

Four controlled trials were conducted to evaluate the therapeutic and persistent efficacy of a new moxidectin formulation (moxidectin 1% nonaqueous injectable) against nematode parasites in cattle. This injectable moxidectin formulation, given as a single subcutaneous injection at a dose rate of 0.02 ml/kg BW to provide 0.2 mg moxidectin/kg BW, was highly efficacious (>90–100%) against larval and/or adult stages of many species of nematodes in cattle including, Dictyocaulus viviparus, Ostertagia spp., Trichostrongylus axei, Haemonchus placei, Trichostrongylus colubriformis, Cooperia spp., Nematodirus helvetianus, Strongyloides papillosus, Oesophagostomum radiatum and Trichuris spp. This formulation had persistent efficacy of >90% against D. viviparus for at least 6 weeks post-treatment, H. placei and Oe. radiatum for 5 weeks post-treatment, and Ostertagia spp. and T. axei for 2 weeks post-treatment.     

Co-inoculation of potassium solubilizing and nitrogen fixing bacteria on solubilization of waste mica and their effect on growth promotion and nutrient acquisition by a forage crop

Waste mica, a potassium-bearing mineral, is a by-product of mica industry; however, its potassium (K)-supplying capacity for crop production is not well understood. A greenhouse trial was made to study the effect of co-inoculation of potassium solubilizing (Bacillus mucilaginosus) and nitrogen (N) fixing (Azotobacter chroococcum A-41) bacteria on solubilization of waste mica (a potassium-bearing mineral) and their effects on growth promotion and nutrient uptake by a forage crop of sudan grass (Sorghum vulgare Pers.) in a Typic Haplustalf. Results revealed that significantly higher biomass accumulation and nutrient acquisition were obtained in all the pots treated with mica and/or bacterial strain as compared to control. Data indicated that co-inoculation of waste mica with B. mucilaginosus and A. chroococcum A-41 resulted in highest biomass production and nutrient acquisition. Co-inoculation of bacterial strains maintained consistently highest amounts of available K and N in soils even at 150 days of crop growth than other treatments. B. mucilaginosus strain was more effective and potent K solubilizer than A. chroococcum A-41. Thus, co-inoculation of potassium solubilizing and nitrogen fixing bacteria to waste mica could be a promising and alternative option for utilizing this potent source as K fertilizer to crops and maintaining greater nutrients availability in soil. Further studies are necessary to see the effects of these bacterial strains on mobilization of potassium-bearing minerals under field conditions.