A novel mannose-binding lectin from plasma of Labeo rohita

A new mannose-binding lectin was purified from plasma of freshwater fish, rohu (Labeo rohita) by ammonium sulphate precipitation followed by DEAE-cellulose ion-exchange chromatography. The hemagglutinating activity is strong for neuraminidase-treated rabbit erythrocytes. Mannose, glucose and their derivatives inhibit hemagglutination. The apparent molecular weight was determined to be 210 kDa in native PAGE. Analysed on SDS-PAGE under reducing and non-reducing conditions, the protein migrated as a single band of mol.wt. 40,000. The lectin is acidic in nature showing a pI of 4.5. It is a glycoprotein containing complex glycan part as indicated by carbohydrate analysis using high-pressure anion exchange chromatography with a pulsed amperometric detector. The N-terminal sequence (WLNGIGTQIPMKITT) shows no significant homology with known proteins. It appears to be a C-type lectin as Ca²⁺ is essential for carbohydrate-binding activity. Methyl -α- D-mannopyranoside was found to be the most potent inhibitor among the monosaccharides and disaccharides tested. Purified lectin was found to induce intracellular superoxide production following opsonization of E. coli by its own macrophages. This revised version was published online in August 2006 with corrections to the Cover Date.     

Soil degradation and mitigation in agricultural lands in the Indian Anthropocene

Current widespread and intensive soil degradation in India has been driven by unprecedented levels of population growth, large-scale industrialization, high-yield agriculture, urban sprawl and the spread of human infrastructure. The damage caused to managed and natural systems by soil degradation threatens livelihoods and local services and leads to national socio-economic disruption. Human-induced soil degradation results from land clearing and deforestation, inappropriate agricultural practices, improper management of industrial effluents and wastes, careless management of forests, surface mining, urban sprawl, and ill-planned commercial and industrial development. Of these, inappropriate agricultural practices, including excessive tillage and use of heavy machinery, over-grazing, excessive and unbalanced use of inorganic fertilizers, poor irrigation and water management techniques, pesticide overuse, inadequate crop residue and/or organic carbon inputs, and poor crop cycle planning, account for nearly 40% (121 Mha) of land degradation across India. Globally, human activities related to agriculture contribute to the transgression of four of the nine Planetary Boundaries proposed by Rockström et al. (2009): Climate Change, Biodiversity Integrity, Land-system Change, and altered Phosphorus and Nitrogen Biogeochemical Flows. This review focuses on how knowledge of soil processes in agriculture has developed in India over the past 10 years, and the potential of soil science to meet the objectives of the United Nations' Sustainable Development Goal 2: Zero Hunger (End hunger, achieve food security, improved nutrition and promote sustainable agriculture), using the context of the four most relevant Planetary Boundaries as a framework. Solutions to mitigate soil degradation and improve soil health in different regions using conservation agricultural approaches have been proposed. Thus, in this review we (1) summarize the outputs of recent innovative research in India that has explored the impacts of soil degradation on four Planetary Boundaries (Climate Change, Biodiversity Loss, Land-system Change, and altered Biogeochemical Flows of Phosphorus and Nitrogen) and vice-versa; and (2) identify the knowledge gaps that require urgent attention to inform developing soil science research agendas in India, to advise policy makers, and to support those whose livelihoods rely on the land.     

Giant piezoelectricity on Si for hyperactive MEMS

Microelectromechanical systems (MEMS) incorporating active piezoelectric layers offer integrated actuation, sensing, and transduction. The broad implementation of such active MEMS has long been constrained by the inability to integrate materials with giant piezoelectric response, such as Pb(Mg(1/3)Nb(2/3))O(3)-PbTiO(3) (PMN-PT). We synthesized high-quality PMN-PT epitaxial thin films on vicinal (001) Si wafers with the use of an epitaxial (001) SrTiO(3) template layer with superior piezoelectric coefficients (e(31,f) = -27 ± 3 coulombs per square meter) and figures of merit for piezoelectric energy-harvesting systems. We have incorporated these heterostructures into microcantilevers that are actuated with extremely low drive voltage due to thin-film piezoelectric properties that rival bulk PMN-PT single crystals. These epitaxial heterostructures exhibit very large electromechanical coupling for ultrasound medical imaging, microfluidic control, mechanical sensing, and energy harvesting.     

Heat shock protein 70 expression in different tissues of Cirrhinus mrigala (Ham.) following heat stress

Members of heat shock protein 70 (HSP70) are highly conserved proteins of about 70 kDa and play important roles in protein folding. Levels of these proteins increase when cells are under stress. Environmental temperature influences both the basal and induced levels of HSPs. However, studies on HSPs in fishes from a tropical country such as India are lacking. In the present study, Indian major carp (IMC) Cirrhinus mrigala (Ham.) acclimatized at 25±2°C had high levels of HSP70, viz., 1.2–1.3 ng μg^?1 total protein in kidney and gill and 4.2–5.3 ng μg^?1 total protein in liver and brain tissues, indicating the presence of biochemically significant levels of stress. However, maintenance of acclimatized fish at 17°C for up to 48 h did not lead to a significant decrease in stress protein levels. A heat shock at 37°C for up to 48 h resulted in only two to threefold increase in HSP70 levels in these organs. Although the increase in HSP70 levels was apparent from the first hour of heat stress in all these tissues, the increase was significant from the second hour in the brain, the sixth hour in liver and kidney and the 20th hour in the gills.     

‘Soya Milk Tris‐based Phytoextender Reduces Apoptosis in Cryopreserved Buffalo (Bubalus bubalis) Spermatozoa’

The objective of this study was to investigate the effect of newly developed soya milk Tris (SMT)‐based phytoextender as an alternative to egg yolk Tris (EYT) extender used for cryopreservation of buffalo (Bubalus bubalis) spermatozoa on apoptosis. Fresh buffalo semen (control without dilution) was cryopreserved in conventional EYT (20% egg yolk v/v in Tris) and SMT (25% soya milk v/v in Tris) extender and used for the assessment of expression of apoptotic proteins. Proteins extracted from a total number of nine ejaculates from three individual buffalo bulls chosen at random were separated using SDS–PAGE followed by immunoblotting against caspase‐8, caspase‐9, caspase‐3, poly(ADP‐ribose)polymerase (PARP), cytochrome c and apoptosis inducing factor (AIF). In addition, fluorescence microscopy was used for the detection of mitochondrial membrane potential (JC‐1 assay) and apoptotic cells (annexin V‐FITC/PI assay). The results obtained clearly indicate the significant (p < 0.05) reduction in the expression of caspase‐3 (27 kDa), caspase‐8 (53 kDa), caspase‐9 (50 kDa) precursor and cytochrome c (17 kDa) in semen cryopreserved in SMT extender in comparison with EYT extender. A non‐significant (p > 0.05) reduction in expression of PARP‐DNA‐binding subunit (24 kDa) was observed in SMT extender. No expression of AIF was found in cryopreserved semen samples. A significant (p < 0.05) increase in the mean percentage of cells having high mitochondrial membrane potential and a non‐significant (p > 0.05) decrease in late apoptotic cells (AN+/PI+) was observed in SMT extender when compared to EYT extender. The results demonstrated that cryopreservation of buffalo semen in SMT‐based phytoextender can replace the traditional egg yolk extenders as it reduces the expression of apoptotic proteins maintaining high mitochondrial membrane potential and gives better protection to sperms in terms of its non‐animal origin.     

Induction,purification and partial characterization of vitellogenin in an Indian major carp Catla catla (Ham.)

Vitellogenin was purified from the serum of 17‐β estradiol (E₂)‐induced juvenile Catla catla using a simple two‐step purification procedure i.e. selective chemical precipitation followed by gel filtration chromatography. Purified protein migrated as single band in a native gradient PAGE which indicated the purity of the sample. The molecular weight of the native catla vitellogenin (~440 kDa) was determined using gel filtration chromatography. In SDS‐PAGE under reducing conditions catla vitellogenin dissociated into three major sub units at 115 kDa, 102 kDa and 73 kDa along with a few faint bands. Confirmation of purified protein as catla vitellogenin was supported by multiple physiological evidences, e.g. absence in male as well as juvenile sera and presence in matured female fish, ability to be synthesized upon estradiol injection in immature fish and certain unique biochemical properties like high molecular weight, phospholipoglycoprotein nature of the molecule. Western blot analysis showed that polyclonal antibody raised against purified protein detected vitellogenin in the sera of catla and in a few species selected from Cyprinidae family. These antisera were used to detect vitellogenin in liver tissue of hormone‐induced catla using immunohistochemistry and its applicability in other immunoassays is discussed.     

Effects of low‐density lipoproteins as additive on quality parameters and oxidative stress following cryopreservation of mithun (Bos frontalis) spermatozoa

Artificial breeding of mithun poses several challenges including lack of standard protocol for cryopreservation of spermatozoa. This is further complicated by harmful effects of hen's egg yolk (EY) as additive in extender. Purified low‐density lipoproteins (LDL) extracted from EY have been shown as beneficial over EY extender for long‐term semen storage in several species. This investigation explored use of LDL versus EY on semen quality and oxidative stress following freezing–thawing of spermatozoa. A total of 25 of 50 ejaculates based on biophysical parameters were selected for the experiment. After diluting with the Tris‐citrate‐glycerol (TCG) extender, each sample was split into three equal aliquots: Group I, control, EY; Group II and Group III contained 8% and 10% purified LDL, respectively. Frozen–thawed samples were evaluated for motility parameters (progressive, and in the bovine cervical mucus penetration test [BCMPT]), viability, sperm and nuclear abnormality, acrosome integrity, and enzymatic (leakage of intracellular contents) and biochemical (oxidative stress) profiles and in vitro fertility (IVF) assay. Study revealed a significant (p < .05) improvement in viability, sperm and nuclear abnormality, acrosome integrity, motility (progressive and in cervical mucus), cholesterol content, and reduction in the leakage of intracellular enzymes in Group II. Moreover, intactness of acrosome and biochemical membranes was protected significantly (p < .05) in addition to significant (p < .05) improvement in binding per cent and binding index in IVF assay in extender containing 8% LDL. These results demonstrate that although cryopreservation of mithun's spermatozoa in EY was comparable with other species, addition of 8% LDL holds a clear advantage over EY or 10% LDL.     

Effect of Supplementation of Taurine or Trehalose in Extender on Immunolocalization of Tyrosine Phosphoproteins in Buffalo and Cattle (Karan Fries) Cryopreserved Spermatozoa

The present study assessed the effects of incorporation of Taurine or Trehalose in extender on immunolocalization of tyrosine phosphoproteins, Cryocapacitation and other sperm quality parameters (motility, viability and membrane integrity) in post‐thawed sperm from Buffalo (Murrah) and Cattle (Karan Fries). Six ejaculates from six individual bulls from both species were chosen at random and split into four aliquots: one aliquot without dilution (fresh sample), another diluted in egg yolk tris‐citrate (EYTC) extender and the rest of aliquots with EYTC dilution supplemented with taurine (50 mm ) or trehalose (100 mm ), respectively, and cryopreserved. Following cryopreservation, semen were thawed and assessed for standard semen quality parameters. Extent of capacitation in cryopreserved spermatozoa was measured by inducing in vitro acrosome reaction followed by dual staining. Immunolocalization of tyrosine phosphoproteins was carried out by immunocytochemistry using primary antibody clone pT‐154 (anti‐phosphotyrosine antibody) and FITC‐conjugated secondary antibody. Immunofluorescent signals were analysed for level of protein tyrosine phosphorylation in spermatozoa. Post‐thaw semen evaluation showed supplementation of taurine or trehalose to EYTC extender significantly (p < 0.05) increased motility, viability and membrane integrity of spermatozoa in both species. Percentage of cryocapacitated spermatozoa was significantly (p < 0.05) higher in cattle as compared to buffalo and degree of cryocapacitaion of spermatozoa decreased significantly (p < 0.05) upon supplementation of additives in both the species. It was also found that tyrosine phosphoproteins were localized differentially in fresh and cryopreserved spermatozoa. Supplementation of taurine or trehalose to freezing extender changed the localization of tyrosine phosphoproteins in cryopreserved spermatozoa similar to fresh in both the species. The results obtained clearly indicated that supplementation of taurine or trehalose to EYTC prior to cryopreservation improves Buffalo and Cattle sperm quality in terms of cryocapacitation and immunolocalization of tyrosine phosphoproteins during freezing–thawing process.     

Outbreaks of egg drop syndrome due to EDS-76 virus in quail (Coturnix coturnix japonica).

Two outbreaks of the egg drop syndrome were observed in quail flocks maintained on a farm together with chickens. The decrease in egg production ranged from 10.6 per cent to 50.6 per cent, and the number of soft-shelled eggs increased with the decline in egg production. In both the outbreaks haemagglutination inhibiting antibodies to EDS-76 virus were detected. Fluorescent viral antigen specific to EDS-76 virus was also detected in the lining epithelial and glandular cells of the uterus.