Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.     

The effect of phosphorus and mulching on the efficiency of phosphorus use and productivity of wheat grown on a mountain Alfisol in the Western Himalayas

Abstract. A field experiment was conducted over two years in one of the mountain Alfisols of the Western Himalayas to study the effects of phosphorus and mulching on phosphorus use efficiency (PUE) and productivity of wheat (cv. S-308) at difference growth stages. The source of phosphorus was single superphosphate at 0, 26, 52 and 78 kg P/ha whereas the sources of mulching materials were pine needles ( Pinus longifolia ), lantana weed ( Lantana camara ) at 8 t/ha and transparent polyethylene sheet compared with no mulch.
The phosphorus use efficiency (PUE) at tillering, flowering and harvesting was greatest at the lowest dose of P and decreased as the P levels were increased. However, the increased levels of P up to 78 kg/ha, significantly improved the dry matter yield at tillering stage (30%) and flowering stage (93%) and also the grain yield (139%) and straw yield (148%) at the harvest stage. The application of mulching materials in general, and polyethylene in particular, significantly increased the phosphorus use efficiency and the productivity of wheat by 27% at tillering stage; 17% at flowering stage, and by 98 and 110% in the case of grain and straw yield at harvest stage.
Although the polyethylene mulch effect was physically superior to the other materials, the latter gave greater financial returns. Consequently, the economic optimum application of P in association with 8 t/ha of lantana mulch was found to be 53 kg/ha over two years, whereas, in the absence of mulching, it was 58 kg/ha.     

Effect of brewer’s yeast on immune response of giant freshwater prawn, Macrobrachium rosenbergii, and its resistance to white muscle disease

The present study was conducted to find the effect of dietary brewer’s yeast on growth, survival, immune response, and resistance to white muscle disease in giant freshwater prawn, Macrobrachium rosenbergii. The brewer’s yeast was supplemented at graded levels 0, 5, 10, and 20?g per kg diet, and the experiment was conducted for 75?days. After the feeding trials, growth, survival, immune parameters like prophenoloxidase activity, respiratory burst, and total hemocyte count were assessed. The growth, specific growth rate, and survival were not found significantly different among the treatment and control groups “prophenoloxidase activity and respiratory burst” were found significantly different (p?<?0.05) among the treatment and control groups. The total hemocyte count was significantly different (p?<?0.05) among control and treatment groups. Cumulative percent survival was higher in M rosenbergii fed 1?% brewer’s yeast-supplemented diet when they were challenged with white muscle disease virus (MrNV (Macrobrachium rosenbergii nodavirus) and XSV (extra small virus)). The present work revealed that the incorporation of brewer’s yeast in the diet is effective in enhancing immune response and controlling the white muscle disease in M rosenbergii.     

An assessment of benzimidazole resistance against caprine nematodes in Central India

Current status of resistance to benzimidazole (BZ) group of anthelmintic drugs against caprine nematodes in Central India at Amanala goat farm, Jabalpur, Madhya Pradesh (M. P.), was systematically investigated using faecal egg count reduction (FECR) test and egg hatch test (EHT). Besides, allele-specific PCR (AS-PCR) was deployed to ascertain the susceptible genotype (alleles) especially of the Haemonchus contortus. Randomly selected 30 goats, irrespective of age and sex, were divided into three groups of 10 each, to serve as treated and untreated controls. It was ensured that the animals were not administered with an anthelmintic drug for the past 3 months prior to undertaking the study, and faecal egg counts were estimated. FECR test evidenced fenbendazole resistance by partial elimination (24.90%) copro-egg counts in the treated group of animals vis-à-vis controls with a lower confidence interval of ?26%. Further, EHT revealed ED-50 value of 0.335 μg of thiabendazole/ml, confirming benzimidazole resistance in the animals of that farm. AS-PCR showed that 62% of H. contortus larvae were homozygous resistant (rr), 24% heterozygous (rS) and 14% homozygous susceptible (SS). The genotypic frequencies of three genotypes (rr, rS and SS) were significantly (P < 0.01) different. The prevalence of benzimidazole resistance allele (r) was also significantly (P < 0.01) higher (74%) as compared to susceptible allele (S) (26%). The resistance to benzimidazole has been discussed while emphasizing improved managemental practices designed to reduce exposure of the goat population to parasites, minimize frequency of anthelmintic use at optimum dose and rotational use of different chemical groups of medicines with different mode of action, so as to overcome and combat the upcoming problem in the field.     

Molecular characterization and variability analysis of <Emphasis Type="Italic">Apple scar skin viroid</Emphasis> in India

Apple scar skin viroid (ASSVd) infection is a major limitation to apple fruit quality and causes huge economic losses. In surveys of apple orchards in the northern Indian state of Himachal Pradesh, fruits with dappling symptoms were noticed. ASSVd was detected from these fruits and molecularly characterized. Ten clones from three isolates were sequenced, of which seven were new sequence variants of ASSVd. The clones had significant sequence variability (94–100%) with each other. Variability was more common in the pathogenic domain of the viroid genome. Four of the clones were 330 nucleotides (nt) long, and the other six had an additional nucleotide. Phylogenetic analysis showed close affinity of the present isolates with some Chinese and Korean isolates. The study reports seven new variants of ASSVd and also provides the first molecular evidence of viroid infection (ASSVd) in apple in India.     

Immunodiagnostic/protective role of cathepsin L cysteine proteinases secreted by Fasciola species

Cathepsin L, a major cysteine proteinase secreted by the parasite plays a pivotal role in various aspects of its pathogenecity. The enzyme takes part in nutrient acquisition by catabolizing host proteins to absorbable peptides, facilitates the migration of the parasite through the host intestine and liver by cleaving interstitial matrix proteins such as fibronectin, laminin and native collagen and is implicated in the inactivation of host immune defenses by cleaving immunoglobulins. Recently, Cathepsin L has been shown to suppress Th1 immune response in infected laboratory animals making them susceptible to concurrent bacterial infections. Accordingly, the protease has been recognized as an important target at which parasite intervention strategies should be directed. Fluke Cathepsin L cysteine proteinases are also reported as sensitive and specific markers for the immunodiagnosis of fasciolosis in ruminants. Further, vaccination of laboratory or large animals with these proteases resulted in a significant reduction in fluke burden and/or fecundity.     

Azadirachtin-A and tetrahydroazadirachtin-A concentrates: preparation, LC-MS characterization and insect antifeedant/IGR activity against Helicoverpa armigera (Hübner)

A 60% azadirachtin-A concentrate has been obtained through repeated precipitation with hexane from a methanolic solution of a 20% concentrate. Azadirachtin-A (90%) has been obtained by medium-pressure liquid chromatography of the 60% concentrate with an RP-18 column and a methanol + water (1 + 1 by volume) solvent system. Catalytic hydrogenation of the 60 and 90% azadirachtin concentrates yielded the corresponding tetrahydroazadirachtin concentrates. Dihydroazadirachtin and tetrahydroazadirachtin formed during the first 5 h of hydrogenation were identified by LC-ESI-MS on the basis of their unique mass fragmentation pattern. The efficacy of tetrahydroazadirachtin concentrates in inhibiting the feeding and growth of Helicoverpa armigera (Hübner) larvae has been compared with that of azadirachtin concentrates. They were in general more active and deterred larvae from feeding at all concentrations. Tetrahydroazadirachtin-A (90%) and azadirachtin-A (90%) with respective IC(50) values of 280 and 390 mg L(-1) were effective as insect growth regulators, while tetrahydroazadirachtin-A (90%) displayed higher antifeedant activity (AI(50) 14 mg L(-1)) against the test insect.     

Adipose triglyceride lipase contributes to cancer-associated cachexia

Cachexia is a multifactorial wasting syndrome most common in patients with cancer that is characterized by the uncontrolled loss of adipose and muscle mass. We show that the inhibition of lipolysis through genetic ablation of adipose triglyceride lipase (Atgl) or hormone-sensitive lipase (Hsl) ameliorates certain features of cancer-associated cachexia (CAC). In wild-type C57BL/6 mice, the injection of Lewis lung carcinoma or B16 melanoma cells causes tumor growth, loss of white adipose tissue (WAT), and a marked reduction of gastrocnemius muscle. In contrast, Atgl-deficient mice with tumors resisted increased WAT lipolysis, myocyte apoptosis, and proteasomal muscle degradation and maintained normal adipose and gastrocnemius muscle mass. Hsl-deficient mice with tumors were also protected although to a lesser degree. Thus, functional lipolysis is essential in the pathogenesis of CAC. Pharmacological inhibition of metabolic lipases may help prevent cachexia.     

Effect of emulsion size and shelf life of azadirachtin A on the bioefficacy of neem (Azadirachta indica A. Juss) emulsifiable concentrates

In a study of 33 recipes of neem oil based emulsifiable concentrates, the specific surface area of the emulsions and cream plus oil layer separation in emulsions at 24 h revealed a correlation of -0.6874 between them and correlations of -0.8940 and 0.6972, respectively, with bioefficacy (LC(50)) against the 3-day-old second-instar larvae of the Bihar hairy caterpillar, Spilosoma obliqua Walker. Nearly 96-99% of azadirachtin A in emulsifiable concentrates (aza-A content = 617.93-1149.65 ppm) degraded during the heat stability test at 54 +/- 1 degrees C for 14 days with half-lives ranging between 1.84 and 4.53 days. The LC(50) values against S. obliqua were, however, statistically at par in both the pre- and the post-heat-treated samples, suggesting a similar effect of azadirachtin A and its degradation products on the bioactivity. The half-life of azadirachtin A could be enhanced by storing the concentrates at lower temperatures. A low pH of the formulation solvent did not check the degradation of azadirachtin A, as reported with aqueous solutions in the literature.