Molecular evolutionary and epidemiological dynamics of a highly pathogenic fish rhabdovirus, the spring viremia of carp virus (SVCV)

Spring viremia of carp virus (SVCV), which causes an acute hemorrhagic and highly contagious disease in cyprinids, was first described in Europe and subsequently reported in parts of Asia and North America. SVCV can be classified into four genogroups: Ia, Ib, Ic, and Id. While Ia and Id have wide circulation and are reported to cause outbreaks in North America and Europe, respectively, Ib and Ic were last reported in the 1980s. We used a Bayesian framework to determine the nucleotide substitution rates, relative genetic diversity, and time to the most recent common ancestor (TMRCA) of SVCV using large sets of sequences of the phosphoprotein and glycoprotein (G) genes. The sampled genetic diversities of Ia and Id were found to have arisen during the year 1996 (95% Highest Posterior Density: 1986-1998) and 1957 (1926-1972), respectively, with consistent results across the two genes. The TMRCA for SVCV was estimated to have been around 1850 (1727-1938). The substitution rate for Ia is at least 5-7 times higher than that of Id. The rate of nonsynonymous (dn) to synonymous (ds) substitutions (dn/ds=ω) for the G gene of Ia (ω=0.608) is significantly higher than that of Id (ω=0.0749), indicating both exhibit distinct selection profiles. The SVCV population experienced a bottleneck during the early 1990 s followed by a sudden rebound, primarily due to the sudden increase in genetic variants in Ia and Id, which coincided with the timing of a recent series of outbreaks reported in Europe and North America.     

Seed variability in <Emphasis Type="Italic">Madhuca longifolia</Emphasis> (Koenig) J. F. Macbride,a source of oil for use as bio-energy

Madhuca longifolia (Koenig) J. F. Macbride is native to India. Seeds contain 20–50% oil (Mahua oil) and the seed cake is used as manure. Mahua oil is used by rural communities for protection against storage pests. Almost all parts of this tree have medicinal properties. Four explorations and collection missions was conducted during July–October 2007 in Tamil Nadu and the Union Territory of Puducherry. This resulted in the collection of 55 accessions. Characterisation and analysis of 7 seed characters were done. The existence of enormous variability in seed length, thickness and 100 seed weight indicates scope for utilization of these accessions for selection of promising material for extraction of oil. Oil content of kernels has been evaluated. Kernel oil ranged from 44.43 to 61.50%. Three accessions IC556617 with 61.50, IC556632 with 60.80 and IC556632 with 60.55% of kernel oil are superior to the rest. Further collections in the site of these oil rich accessions are promising for more productive Madhuca longifolia genetic resources.     

High antigenic cross-reaction among the bacterial species responsible for diseases of cultured freshwater fishes and strategies to overcome it for specific serodiagnosis

Antigenic sharing among the most commonly bacterial pathogens such as Aeromonas hydrophila, Edwardsiella tarda and Pseudomonas fluorescens of Indian major carps has been studied using immunological reactions such as cross-agglutination, disc diffusion and indirect enzyme linked immunosorbent assay (ELISA). The data were analysed using statistical analysis (SAS), version 6.12. The results showed high antigenic similarities among the bacterial whole cells, whole cell lysates, somatic 'O' antigens, lipopolysaccharides (LPS) and extracellular products (ECP). However, few or no similarities were observed in ECP components of <20kD. The present study indicates a need to develop differential diagnostic methods based on serology by choosing the highly specific less cross-reactive ECP antigen.     

High-level expression of SAG1 and GRA7 gene of Toxoplasma gondii (Izatnagar isolate) and their application in serodiagnosis of goat toxoplasmosis

Toxoplasmosis, caused by Toxoplasma gondii, is a disease of economic importance in livestock, especially in sheep and goats, where it causes abortion. Although several serological tests are in use for diagnosis of infection, production of reliable reagents is a constraint. An 814 bp sequence coding for a truncated surface antigen surface antigen 1 (SAG1), a tachyzoite stage-specific protein, as well as a 657 bp sequence coding for granule protein 7 (GRA7), a dense granule protein were PCR amplified from the genomic DNA of T. gondii. The amplified products were ligated in pET-32b(+) and pET-32c(+) expression vectors, respectively and subsequently transformed into BL21(DE3)pLysS cells. A high-level expression of the histidine-tagged SAG1 and GRA7 fusion proteins were obtained after 7h of incubation. The recombinant proteins were purified using Ni-NTA column and were characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis using reference positive sera from goat, rabbit and humans at 1:100 dilution. Subsequently, the diagnostic efficiency of the recombinant proteins, either individually or as a cocktail of the recombinant proteins, was assessed with 56 reference goat sera by enzyme-linked immunosorbent assay (ELISA). The immunoreactivity of the refolded SAG1 and GRA7 was evidenced by high OD values. The reactivity of the recombinant proteins as a cocktail preparation was more than that of individual proteins in ELISA and could detect accurately the infection in goats. This is the first report of serological detection of caprine toxoplasmosis by ELISA using a cocktail of recombinant Toxoplasma proteins.     

Changes in Plasma Concentrations of LH, FSH, Estradiol 17-β and Progesterone During Oestrus in Mithun (Bos frontalis)

The objective of the present study was to establish the changes in plasma concentrations of LH, FSH, estradiol 17-beta (E2) and progesterone (P4), as well as to understand their temporal relationships during oestrus in mithun (Bos frontalis). The experiment was conducted on 11 mithuns during third or fourth postpartum oestrous cycle. Since oestrus onset the jugular vein blood samples were collected every 2 h for 72 and 96 h, respectively from the animals without and with standing heat. The LH, FSH, E2 and P4 concentrations were estimated in plasma. The P4 concentration was fluctuated throughout the oestrus period and the average P4 concentration was found significantly (p<0.05) lower on the day of oestrus onset. The multiple rises in LH and FSH concentrations above the basal level in spike like fashion were observed throughout the oestrus period irrespective of the occurrence of standing heat. A significant (p<0.01) gradual increase in the average daily E2 concentration was observed till day 2 following oestrus onset irrespective of the occurrence of standing heat. A significant (p<0.05) simultaneous increase in LH, FSH and E2 concentrations and a transient increase in P4 concentration at approximately the time of standing heat onset were observed. During investigation a definite temporal coupling between LH and FSH rises was absent throughout the oestrus period. The results suggest that (1) the multiple short-duration low-amplitude LH and FSH surges during oestrus may be crucial for the final maturation of ovulatory follicle and subsequent ovulation in mithun; (2) a differential mechanism for controlling LH and FSH secretions probably exists in mithun.     

Goat serum, a substitute of bovine serum in cultivation of Babesia bovis.

Babesia bovis (a Mexican isolate) was cultivated in MASP culture system using goat serum in various concentrations as substitute of bovine serum. It was observed that 20% goat serum + 20% bovine serum + 60% Parker's medium 199 supported the growth of the parasite, which was maintained in this medium through 8 subcultures. The soluble exoantigen (vaccine) present in the culture supernatant is to be quantified and tested in vitro. Goat serum from slaughterhouses may be utilized for in vitro cultivation of the parasite and, expectedly, production of vaccine. This study may prove to be useful in reducing the cost of vaccine at least in tropical countries.     

Genetic typing of bovine viral diarrhoea virus isolates from India

Thirteen BVDV isolates collected in four geographic regions of India between 2000 and 2002 were typed in 5'-UTR. To confirm results of genetic typing, selected viruses were also analysed in the N(pro) region. Phylogenetic analysis revealed that all Indian BVDV isolates belong to BVDV-1b (Osloss-like group). Despite a long distance between the farms from which the viruses were isolated there was no correlation between the origin of viral isolates and their position in a phylogenetic tree. Higher genetic similarity of Indian BVDV isolates was observed most probably due to the uncontrolled movement of cattle as well as the uncontrolled use of semen from bulls for breeding of local and farm cattle in different states of India.     

Marker-assisted breeding to develop the drought-tolerant version of Sabitri,a popular variety from Nepal

Sabitri is a rice variety grown in a large part of the rainfed areas of Nepal. It was originally developed for irrigated condition; hence, this variety suffers high yield decline under drought. Two QTLs, qDTY _3.2 and qDTY _12.1 , with large effects on grain yield under drought were identified in the Sabitri background in separate QTL mapping studies. The present study reports the development of Sabitri near isogenic lines (NILs) with combinations of these two QTLs and their characterization under drought. To do so, marker-assisted backcross breeding (MABB) was combined with phenotypic selection to develop high-yielding drought-tolerant NILs with Sabitri grain type. Apart from this, drought-tolerant variants for grain type with high yield under non-stress were identified among the developed NILs. Early days to flowering of up to 13 days and reduction in plant height of up to 13 cm as compared to Sabitri were observed in the developed NILs. Some of these NILs showed higher yield compared to Sabitri and relatively higher tolerance to drought, indicating the capture of positive alleles and interactions during the course of selection. The developed NILs possessed high yield potential which make them suitable materials for the testing of water-saving technologies in irrigated areas. Based on their performance, these NILs can be deployed in rainfed areas in Nepal and other countries of South Asia to increase yield stability.     

Basalt nanoparticle reinforced hybrid woven composites: Mechanical and thermo-mechanical performance

Nanoparticles are gaining wider importance and increasing utility in many areas of engineering and technology. This investigative work is conducted to study the effect of incorporating basalt nano particles in composites with basalt/basalt and basalt/jute woven structures as reinforcement. The nanoparticles were developed from basalt, they were characterized and used for reinforcing composites of basalt and jute hybrid fabrics. The mechanical and thermo-mechanical properties of hybrid woven basalt reinforced epoxy composites were evaluated. Microscopic examination was carried out in order to analyze the internal structure and fractured surfaces. Interfacial characteristics, material morphology and failure was studied by use of Scanning Electron Microscope (SEM) and optical microscopy. Thermal stability was characterized by TGA. The results elaborated that the incorporation of basalt nanoparticles exhibited superior properties compared to the pure epoxy resin impregnated basalt fabric reinforced composites in terms of mechanical and thermal stability.