One-step RT-PCR for the Detection of Infectious Bursal Disease Virus in Clinical Samples

A single-tube, non-interrupted, one-step RT-PCR has been standardized to amplify the hypervariable region of the VP2 gene sequence of infectious bursal disease virus (IBDV). The technique standardized on purified viral RNA was successfully applied to the detection of the virus directly in clinical samples. The amplified products were confirmed to be IBDV specific by their size in ethidium bromide-stained agarose gel, nested PCR and restriction enzyme digestion. Digestion of the amplicons with StyI restriction enzyme also differentiated classical virus from six very virulent field isolates. The sensitivity of the one-step RT-PCR was found to be 0.2 pg of viral RNA.     

Evaluation of pulp and paper making characteristics of certain fast growing plants

Summary Four species of fast growing annual plant viz.Hibiscus sabdariffa, Crotalaria juncea, Tephrosia candida andHibiscus cannabinus and a variety of reed, Neyraudia reynaudiana were evaluated in the laboratory for their pulp and paper making properties. Data on proximate chemical analysis of raw materials, unbleached and bleached pulp properties, morphological properties of fibres and physical strength properties of paper sheets were evaluated. The unbleached pulp yields were 44.90–53.20% with all the five plant materials, while bleached pulp yields varied from 39.80–50.60%. The average fibre lengths and diameters of the unbleached pulp obtained from the five plant species were within the range of 0.75 mm–2.15 mm and 20–22 pun respectively. Paper hand sheets, made with bleached pulps (45° SR) obtained from the five plant species, gave burst indices 2.75–3.92 kPa m²/g, tear indices 3.73–9.61 m N m²/g and tensile indices 37.27–47.81 N mg^–1, indicating adequate strength properties for writing and printing type of paper.The authors thank Director, Regional Research Laboratory (CSIR), Jorhat, India, for his kind permission to publish this paper. The authors also wish to thank B. N. Tosh, JPF for his assistance.     

Performance of F1 progenies developed through crosses between Brassica carinata A. Braun A(♀) and Brassica rapa L (♂)

Genetic Resources and Crop Evolution - The present investigation was aimed to study the performance of F1 progenies of crosses between Brassica carinata A. Braun and Brassica rapa L. considering...     

Formulation of cheaper artificial feeds for shrimp culture: Preliminary biochemical,physical and biological evaluation

A compound shrimp diet was prepared making use of the locally available natural food stuffs, slaughter house products and factory by-products. The ingredients, viz: beef liver (discarded for human use on veterinary grounds), coconut oil cake, soya flour, mangrove leaves, and wheat flour, were analysed biochemically for their nutritive value. The formulated diet is in a dry pellet form. It was assessed experimentally for its physical stability and subsequently fed to various size groups of a palaeomonid prawn (Leptocarpus potamiscus) for biological evaluation. The results of the investigations suggest that beef liver meal and coconut oil cake are rich in protein and can be utilized in the formulation of aquatic diets. The diet is highly water stable, consumable and assimilable. It should be tested upon both palaeomonid and penaeid prawns in further experiments.     

Pathogenicity and In Planta Mycotoxin Accumulation Among Members of the Fusarium graminearum Species Complex on Wheat and Rice

ABSTRACT Fusarium head blight (FHB), or scab, is a destructive disease of small grains caused by members of the Fusarium graminearum species complex, comprised of at least nine distinct, cryptic species. Members of this complex are known to produce mycotoxins including the trichothecenes deoxynivalenol (DON) along with its acetylated derivatives and nivalenol (NIV). In this study, 31 strains, belonging to eight species of this complex and originating from diverse hosts or substrates, were tested for differences in aggressiveness and mycotoxin production. Large variation among strains, both in terms of their aggressiveness and the ability to produce trichothecenes on a susceptible cultivar of wheat was found; variation appears to be a strain-specific rather than species-specific characteristic. While pathogenicity was not influenced by the type of mycotoxin produced, a significant correlation was observed between the amount of the dominant trichothecene (DON and its acetylated forms or NIV) produced by each strain and its level of aggressiveness on wheat. Some isolates also were tested for their ability to infect rice cv. M201, commonly grown in the United States. While tested strains were capable of infecting rice under greenhouse conditions and causing significant amount of disease, no trichothecenes could be detected from the infected rice florets.     

Marker‐assisted introgression of rare allele of β‐carotene hydroxylase (crtRB1) gene into elite quality protein maize inbred for combining high lysine,tryptophan and provitamin A in maize

Vitamin A deficiency in humans is a widespread health problem. Quality protein maize (QPM) is a popular food rich in lysine and tryptophan, but poor in provitamin A (proA). Here, we report the improvement of an elite QPM inbred, HKI1128Q for proA using marker‐assisted introgression of crtRB1‐favourable allele. HKI1128 was one of the parental lines of three popular hybrids in India and was converted to QPM in our earlier programme. Severe segregation distortion for crtRB1 was observed in BC₁F₁, BC₂F₁ and BC₂F₂. Background selection by 100 SSRs revealed mean recovery of 91.07% recurrent parent genome varying from 88.78% to 93.88%. Across years, introgressed progenies possessed higher mean β‐carotene (BC: 9.22 µg/g), β‐cryptoxanthin (BCX: 3.05 µg/g) and provitamin A (proA: 10.75 µg/g) compared to HKI1128Q (BC: 2.26 µg/g, BCX: 2.26 µg/g and proA: 3.38 µg/g). High concentration of essential amino acids, viz. lysine (mean: 0.303%) and tryptophan (0.080%) in endosperm, was also retained. Multi‐year evaluation showed that introgressed progenies possessed similar grain yield (1,759–1,879 kg/ha) with HKI1128Q (1,778 kg/ha). Introgressed progenies with higher lysine, tryptophan and proA hold immense potential as donors and parents in developing biofortified hybrids.     

Relative potencies of natural estrogens on vitellogenin and choriogenin levels in the Indian freshwater spotted snakehead, <Emphasis Type="Italic">Channa punctata</Emphasis>: in vivo and in vitro studies

The relative efficacies of three natural estrogens viz., estrone (E₁), estradiol-17β (E₂) and estriol (E₃) to induce synthesis of vitellogenin (Vg) and choriogenin (Chg) were assessed in primary hepatocyte cultures of the Indian freshwater spotted snakehead, Channa punctata. Hepatocytes were isolated from the spotted snakehead liver by a non-enzymatic protocol. Optimum culture conditions were standardized for ensuring their viability and functioning. Isolated hepatocytes were cultured for 48 h for monolayer formation and then exposed to various concentrations (0.001–10 μM) of the three estrogens. Competitive homologous ELISAs, developed and validated for spotted snakehead Vg and Chg were employed to determine the amounts of these two proteins secreted into the culture medium after 48 h of incubation. The results reveal that although all the three estrogens were effective in inducing the production of Vg and Chg in a dose-dependent manner, there were differences in their relative potencies. Of three estrogens, E₁ was the least potent and could induce synthesis of Vg and Chg only at a minimum concentration of 0.5 μM; whereas significant levels of both the proteins were quantified in culture medium by exposing the hepatocytes to E₂ or E₃ even at a concentration of 0.001 μM. All three estrogens were effective in inducing synthesis of Vg and Chg in vivo also. These results suggest the possibility of employing the above in vitro experimental design to monitor the presence of estrogens/estrogen-like chemicals in natural waters, which could interfere with the estrogen receptor system of fish. This study further points to the possibility of using Chg, in addition to Vg, as a parameter for screening various chemicals for their estrogenic activity.     

Development and characterization of a cell line TTCF from endangered mahseer Tor tor (Ham.)

Tor tor is an important game and food fish of India with a distribution throughout Asia from the trans-Himalayan region to the Mekong River basin to Malaysia, Pakistan, Bangladesh and Indonesia. A new cell line named TTCF was developed from the caudal fin of T. tor for the first time. The cell line was optimally maintained at 28°C in Leibovitz-15 (L-15) medium supplemented with 20% fetal bovine serum (FBS). The propagation of TTCF cells showed a high plating efficiency of 63.00%. The cytogenetic analysis revealed a diploid count of 100 chromosomes at passage 15, 30, 45 and 60 passages. The viability of the TTCF cell line was found to be 72% after 6 months of cryopreservation in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 578- and 655-bp sequences of 16S rRNA and cytochrome oxidase subunit I (COI) genes of mitochondrial DNA (mtDNA) respectively. TTCF cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids. Further, immunocytochemistry studies confirm its fibroblastic morphology of cells. Genotoxicity assessment of H?O? in TTCF cell line revealed the utility of TTCF cell line as in vitro model for aquatic toxicological studies.