Alterations in the epidermis of the carp,Labeo rohita (Cyprinidae: Cypriniformes), infected by the bacteria,Aeromonas hydrophila: A scanning electron microscopic,histopathological and immunohistochemical investigation

This study was carried out to comprehend the pathogenicity of the bacteria in the epidermis of Labeo rohita inoculated with Aeromonas hydrophila. Alterations in the histopathology of the epidermis were examined using scanning electron microscopy, light microscopy and the localization of iNOS and caspase 3 + ve cells by means of immunohistochemical methods. Skin samples obtained from infected fish at different intervals 2, 4, 6, 8 and 10 days showed significant changes in the cellular components of the epidermis. Epithelial cells often appeared hypertrophied with fragmented and loosely arranged microridges, and in the process of exfoliation. Mucous goblet cells increased significantly in density. Club cells showed degenerative changes, often with simultaneous confluence of adjacent cells and release of their contents. Increase in density of iNOS and caspase 3 + ve cells indicates inflammatory response and apoptosis. This study could provide valuable information on the pathogenesis of the disease, and disease outbreaks in farmed fish. Further, it could provide useful guidelines for fish farmers to take preventive measures for the control of the disease.     

Evaluation of antibacterial activity and innate immune components in skin mucus of Indian major carp,Cirrhinus mrigala

The present work has been undertaken to analyse the antibacterial activity and innate immune components in the skin mucus of Indian major carp, Cirrhinus mrigala. Skin mucus was extracted separately in triple‐distilled water (TDW), 3% acetic acid (3% AA) or 0.1% trifluoroacetic acid (1% TFA). All mucus extracts exhibited different spectrum of the antibacterial activity against different groups of pathogenic bacteria. Protein profiling by polyacrylamide gel electrophoresis revealed a series of protein bands in the TDW extract, four major protein bands in the AA extract and two protein bands in the TFA extract. Tandem mass spectrometry analysis of distinct protein bands identified potential innate immune factors – histone H2A, histone H3, histone H4, haemoglobin, cofilin and nucleoside diphosphate kinase in the TDW extract, and ubiquitin and histone H2B isoforms in acidic extracts of skin mucus of C. mrigala. The presence of these innate immune molecules suggests that skin mucus play an important role in the protection of the fish against microbial invasion.     

Isolation and cultivation of equine corneal keratocytes, fibroblasts and myofibroblasts

Objective  To establish an in vitro model for the investigation of equine corneal wound healing. To accomplish this goal, a protocol to isolate and culture equine corneal keratocytes, fibroblasts and myofibroblasts was developed.
Animal material  Equine corneal buttons were aseptically harvested from healthy research horses undergoing humane euthanasia for reasons unrelated to this study. Slit-lamp biomicroscopy was performed prior to euthanasia by a board-certified veterinary ophthalmologist to ensure that all samples were harvested from horses free of anterior segment disease.
Procedure  Equine corneal stroma was isolated using mechanical techniques and stromal sub-sections were then cultured. Customized media at different culture conditions was used to promote growth and differentiation of corneal stromal cells into keratocytes, fibroblasts and myofibroblasts.
Results  Cell culture techniques were successfully used to establish a method for the isolation and culture of equine corneal keratocytes, fibroblasts and myofibroblasts. Immunohistochemical staining for alpha-smooth muscle and F-actin was used to definitively differentiate the three cell types.
Conclusion  Equine corneal stromal keratocytes, fibroblasts and myofibroblasts can be predictably isolated and cultured in vitro using this protocol.     

Induction of systemic resistance to <Emphasis Type="Italic">Exobasidium vexans</Emphasis> in tea through SAR elicitors

The effect of two chemical elicitors (acibenzolar-S-methyl benzo-[1,2,3]- thiadiazole-7-carboxylic acid S-methyl ester [Boost 500SC]) and salicylic acid in inducing resistance in tea plants against blister blight disease caused by Exobasidium vexans Massee, was studied. Treatments with elicitors resulted in reduced severity of blister blight disease in nursery plants on challenge with the pathogen. There was a significant increase in the activities of defense enzymes like phenylalanine ammonia lyase, peroxidase and β-1,3-glucanase on elicitor treatments in tea leaves challenged with the pathogen than on unchallenged leaves. Acibenzolar-S-methyl (ASM) at 0.14% registered the lowest disease severity (25.2%), whereas treatments with salicylic acid were inferior. Under field conditions, the application of ASM at 0.14% resulted in disease protection of 25%. When ASM was applied in alternate rounds with a standard fungicide, the disease protection improved to 46.8%. The importance of incorporating ASM as a component in integrated disease management and also its importance in organic tea cultivation is discussed.     

Validation and Marker-Assisted Selection of Stem Rust Resistance Gene Sr2 in Indian Wheat Using Gel-Based and Gel-Free Methods

Journal of Crop Science and Biotechnology - Stem rust resistance gene Sr2 is an important slow-rusting gene which has provided resistance against stem rust for many years. The Sr2 gene has durable...     

Marker-assisted introgression of QTLs for yield under moisture stress into elite varieties of rice (Oryza sativa)

Rice production is severely constrained by the moisture stress. The present study was undertaken to transfer two important QTLs viz., (qDTY2.2 and qDTY4.1), which controls yield under moisture stress (DTY) into two elite varieties, that is, MTU1010 and NLR34449 through marker-assisted breeding. Foreground and background selections of backcross generations, that is, BC₁F₁, BC₂F₁ and BC₂F₂ identified several promising introgression lines (ILs) with two QTLs and single QTLs with an appreciable level of recovery of recipient parent genome. In ILs, the recovery of MTU1010 genome content was estimated to be 83%–93% while the recovery of NLR34449 was 84%–92%. The two-QTL ILs of MTU1010 and NLR34449 backgrounds (qDTY2.2 and qDTY4.1) have shown substantial yield advantage (32 to 84%) over the single-QTL ILs (either qDTY2.2 or qDTY4.1) under moisture stress conditions. Among all, two ILs, MBC-124 and NBC-127 are the best high yielding lines under moisture stress conditions. These outyielded ILs have the potential to be released as varieties in rainfed ecosystem and also can be used as donors in the existing breeding programme in rice.     

Effects of Varying Concentrations of l-carnitine-incorporated Diets on Growth and Body Composition of Fry of Cirrhinus mrigala (Hamilton, 1822)

Experiment was conducted to ascertain the effect of l ‐carnitine on growth and body composition of Cirrhinus mrigala fry (0.342 ± 0.03 g) using five different concentrations (0.25, 0.50, 0.75, 1, and 0%) of l ‐carnitine‐incorporated diets for 120 d. At the end of trial, weight gain, feed conversion efficiency, and protein efficiency of fishes fed with 0.25% l ‐carnitine diet were observed to be significantly higher. Poor growth was recorded in the fry fed diet with no carnitine, while intermediate growth was observed when fed with 0.50, 0.75, and 1% l ‐carnitine‐incorporated diets. The analysis of body composition of C. mrigala fry indicated that there is no significant difference (P > 0.05) in moisture, protein, and ash contents, but significant difference (P < 0.05) was found in body lipid content of fry, fed with different concentrations of l ‐carnitine. Decrease in viscerosomatic and hepatosomatic indices were observed in fishes fed with l ‐carnitine‐incorporated diets. Results of this study indicated that diet containing 0.25% l ‐carnitine can promote higher growth in C. mrigala fry.     

The BamHI site in the Internal Transcribed Spacer Region of Mungbean ribosomal RNA gene is partially methylated

The 18s-5.8s-25s ribosomal gene (18s-25s rDNA) in higher plants is present in multiple copies, on different chromosomes, as tandemly repeated units. Among the multiple BamHI sites that occur in the repeat unit, only the site in the middle of the 25s rRNA coding region is methylated in most cereals and pulses. BamHI restriction enzyme analyses of the mungbean 18s-25s rDNA showed the presence of two populations. Nearly 50% of the 18s-25s rDNA population had BamHI site situated in the internal transcribed spacer region (ITS-BamHI) resistant to cleavage by BamHI. The amplified ITS fragment was completely digestible by BamHI showing that partial cleavage by BamHI is not due to variation in the recognition sequence but most probably due to methylation. The complete cleavage of the ITS-BamHI site by BstYI that recognizes BamHI site but is insensitive to methylation confirms that the ITS-BamHI site is methylated. Methylation is probably due to the presence of a guanosine residue adjacent to the 3′ cytosine in the recognition sequence. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of the root zone water quality model for predicting water and NO3–N movement in an Iowa soil

Evaluation of computer models with field data is required before they can be effectively used for predicting agricultural management systems. A study was conducted to evaluate tillage effects on the movement of water and nitrate–nitrogen (NO₃–N) in the root zone under continuous corn (Zea mays L.) production. Four tillage treatments considered were: chisel plow (CP), moldboard plow (MP), no-tillage (NT), and ridge-tillage (RT). The root zone water quality model (RZWQM: V.3.25) was used to conduct these simulations. Three years (1990–1992) of field observed data on soil water contents and NO₃–N concentrations in the soil profile were used to evaluate the performance of the model. The RZWQM usually predicted higher soil water contents compared with the observed soil water contents. The model predicted higher NO₃–N concentrations in the soil profile for MP and NT treatments in comparison with CP and RT treatments, but the magnitude of simulated NO₃–N peak concentrations in the soil profile were substantially different from those of the observed peaks. The average NO₃–N concentrations for the entire soil profile predicted by the model were close to the observed concentrations except for ridge tillage (percent difference for CP=+5.1%, MP=+12.8%, NT=+18.4%, RT=−44.8%). Discrepancies between the simulated and observed water contents and NO₃–N concentrations in the soil profile indicated a need for the calibration of plant growth component of the model further for different soil and climatic conditions to improve the N-uptake predictions of the RZWQM.     

Wound‐healing potential of curcumin in the carp,Labeo rohita

In this investigation, the effect of dietary administration of curcumin on the healing of skin wound in fish, Labeo rohita, has been reported. Fish were divided into three groups: control group (fish without skin wound), sham group (fish with skin wound without curcumin treatment) and curcumin‐treated group (fish with skin wound and subjected to dietary administration of 1% curcumin). Experiments were conducted for 30 days to assess the healing of skin wounds at different time intervals using scanning electron microscopy, histology, and mucopolysaccharide and enzyme histochemistry. In the curcumin‐treated group, healing of skin wounds was found to be enhanced than in the sham group as indicated by early restoration of morphology of the surface layer of epithelial cells; the density of the mucous goblet cells; the density of club cells in epidermal layer; and early granular tissue formation, collagen deposition and tissue remodelling in dermal layer. Furthermore, peroxidase and catalase enzyme activity showed increased endogenous defence system in the curcumin‐treated group compared with the sham group. It could be concluded that dietary administration of curcumin is beneficial in rapid healing of skin wounds in fish. Early healing of wounds could be considered to prevent the invasion of pathogens and to maintain the integrity of the surrounding tissue.