New metabolites in the degradation of alpha- and gamma-hexachlorocyclohexane (HCH): pentachlorocyclohexenes are hydroxylated to cyclohexenols and cyclohexenediols by the haloalkane dehalogenase LinB from Sphingobium indicum B90A

Technical hexachlorocyclohexane (HCH) and lindane are obsolete pesticides whose former production and use led to widespread contaminations posing serious and lasting health and environmental risks. Out of nine possible stereoisomers, alpha-, beta-, gamma-, and delta-HCH are usually present at contaminated sites, and research for a better understanding of their biodegradation has become essential for the development of appropriate remediation technologies. Because haloalkane dehalogenase LinB was recently found responsible for the hydroxylation of beta-HCH, delta-HCH, and delta-pentachlorocyclohexene (delta-PCCH), we decided to examine whether beta- and gamma-PCCH, which can be formed by LinA from alpha- and gamma-HCH, respectively, were also converted by LinB. Incubation of such substrates with Escherichia coli BL21 expressing functional LinB originating from Sphingobium indicum B90A showed that both beta-PCCH and gamma-PCCH were direct substrates of LinB. Furthermore, we identified the main metabolites as 3,4,5,6-tetrachloro-2-cyclohexene-1-ols and 2,5,6-trichloro-2-cyclohexene-1,4-diols by nuclear magnetic resonance spectroscopy and gas chromatography-mass spectrometry. In contrast to alpha-HCH, gamma-HCH was not a substrate for LinB. On the basis of our data, we propose a modified gamma-HCH degradation pathway in which gamma-PCCH is converted to 2,5-cyclohexadiene-1,4-diol via 3,4,5,6-tetrachloro-2-cyclohexene-1-ol and 2,5,6-trichloro-2-cyclohexene-1,4-diol.     

Untapped amaranth (<Emphasis Type="Italic">Amaranthus </Emphasis>spp.) genetic diversity with potential for nutritional enhancement

Significant genetic diversity was observed in 218 out of a total of 1309 accessions of amaranth (Amaranthus hypochondriacus L.) and its seven wild relatives, A. spinosus L., A. dubius Mart. ex Thell., A. hybridus L., A. tricolor L., A. cruentus L., A. caudatus L., A. retroflexus L. for 24 nutritional parameters including total oil content, fatty acid profile, total protein content and amino acid profile. Diversity for total oil content (6.42–12.53%), linoleic acid (25.68–54.34%), oleic acid (21.97–42.01%) of the total fatty acids, total protein content (7.84–18.01%), among important essential amino acids; lysine content (0.66–11.12 g/16 g N), methionine (0.35–4.80 g/16 g N) and half cystine and (0.12–8.32 g/16 g N) was reported. The un-weighted pair-group method using arithmetic average cluster analysis based on pair wise Euclidean genetic distance grouped the accessions into seven major clusters. Histidine, half cystine, tyrosine, essential amino acids, total oil content, linoleic acid and oleic acid content were the major parameters contributing significantly to genetic diversity. Present findings indicate that significant diversity exists for nutritional parameters in amaranth germplasm. The promising accessions with higher multiple nutritive traits; protein content (>16%), oil content (>11%), lysine content (>7.5 g/16 g N) and EAA higher than the FAO reported values, were identified. This is the first report on detailed nutritional analysis of diversity collected worldwide. These could be used as potential breeding material for nutritional enhancement through genetic improvement. This will help in overcoming the “triple burden” of malnourishment, hidden hunger, and obesity.     

Nutrient Management for Sustaining Productivity of Sunflower-Based Cropping Sequence in Indian Semiarid Regions

A comprehensive long-term study (2006–2010) was undertaken to develop a balanced and integrated nutrient supply system for sunflower-based cropping sequence considering the efficient utilization of residual and cumulative soil nutrient balance along with added fertilizers by the crops grown in rotation. The fertilizer application was done in potato and sunflower while greengram was raised as such on their residual effect. Significant response in yield was observed with 150% of the recommended nitrogen, phosphorus and potassium (NPK) or inclusion of farmyard manure (FYM) with the recommended NPK in the cropping sequence indicating 6.2–7.0% gain in system productivity over the existing recommendations. Each additional unit of P and K nutrition prompted system productivity by 18.9 and 11.0 kg kg^?1 of applied nutrient, respectively. Apparent yield decline was observed in K and PK omission plots to the extent of 15.8 and 27.4% in potato, 10.5 and 23.9% in sunflower and 4.2 and 8.3% in greengram, respectively, compared to the recommended fertilization. The superiority of the FYM along with the recommended NPK (potato/sunflower) was evident on the overall profitability and sustainability of the system, highlighted by the significantly higher productivity (7.16 t SFEY ha^?1), sustainability yield index (SYI; 0.76), production efficiency (PE; 27.85 kg SFEY ha^?1 day^?1) and net returns (2520 USD ha^?1) with a B:C ratio of 2.91. Apparent change in potassium permanganate (KMnO₄)-N was negative in all the treatments while N and P balance was positive with 150% NPK fertilization. Nutrient uptake exceeded the replenishment with 100% NPK application and maintained net negative soil nutrient stock for all the primary nutrients, indicating the need for revalidation of the existing recommendations in the system perspective. Conspicuous improvement in residual soil fertility in terms of maximum buildup of soil organic carbon (14%) and enhancement in soil KMnO₄-N (4.2%), Olsen-P (19.4%), ammonium acetate (NH4OAc)-K (5.8%) and dehydrogenase enzyme activity (44.4%) was observed in FYM-treated plots over the initial values. The study suggested that the inclusion of legumes and FYM application with the recommended NPK in potato-sunflower cropping sequence will sustain the system’s productivity through the efficient use of nutrients, enhanced microbial activity and improved soil health while combating escalating prices of fertilizers as well as environmental issues in the Indo-Gangetic plains of India and similar environments.     

Complete genome sequence analysis of chicken astrovirus isolate from India

Objective

Chicken astroviruses have been known to cause severe disease in chickens leading to increased mortality and “white chicks” condition. Here we aim to characterize the causative agent of visceral gout suspected for astrovirus infection in broiler breeder chickens.

Methods

Total RNA isolated from allantoic fluid of SPF embryo passaged with infected chicken sample was sequenced by whole genome shotgun sequencing using ion-torrent PGM platform. The sequence was analysed for the presence of coding and non-coding features, its similarity with reported isolates and epitope analysis of capsid structural protein.

Results

The consensus length of 7513 bp genome sequence of Indian isolate of chicken astrovirus was obtained after assembly of 14,121 high quality reads. The genome was comprised of 13 bp 5′-UTR, three open reading frames (ORFs) including ORF1a encoding serine protease, ORF1b encoding RNA dependent RNA polymerase (RdRp) and ORF2 encoding capsid protein, and 298 bp of 3′-UTR which harboured two corona virus stem loop II like “s2m” motifs and a poly A stretch of 19 nucleotides. The genetic analysis of CAstV/INDIA/ANAND/2016 suggested highest sequence similarity of 86.94% with the chicken astrovirus isolate CAstV/GA2011 followed by 84.76% with CAstV/4175 and 74.48%% with CAstV/Poland/G059/2014 isolates. The capsid structural protein of CAstV/INDIA/ANAND/2016 showed 84.67% similarity with chicken astrovirus isolate CAstV/GA2011, 81.06% with CAstV/4175 and 41.18% with CAstV/Poland/G059/2014 isolates. However, the capsid protein sequence showed high degree of sequence identity at nucleotide level (98.64-99.32%) and at amino acids level (97.74–98.69%) with reported sequences of Indian isolates suggesting their common origin and limited sequence divergence. The epitope analysis by SVMTriP identified two unique epitopes in our isolate, seven shared epitopes among Indian isolates and two shared epitopes among all isolates except Poland isolate which carried all distinct epitopes.

     

Somatic hypermutation leads to diversification of the heavy chain immunoglobulin repertoire in cattle

The availability of unique variable (VH), diversity (D), and joining (JH) gene segments in the vertebrate germline determines the extent to which a primary immunoglobulin (Ig) repertoire can be generated through combinatorial rearrangement. Although bovine D segments possess unusual properties, the diversity of the primary Ig heavy chain (IgH) repertoire in cattle is restricted by the dominance of a single family of germline VH genes of limited number and diversity. Cattle therefore must employ other diversification strategies in order to generate a functional IgH repertoire, the main candidates being gene conversion and somatic hypermutation. In considering these possibilities, we predicted that if somatic hypermutation was active during B lymphocyte development, the process would introduce nucleotide substitutions to the VDJ exon and also non-coding region lying downstream of the rearranged JH segment. In contrast, our expectation was that gene conversion would show a greater tendency to confine modification to the IgH coding sequence, leaving intron regions substantially unmodified. An analysis of rearranged IgH sequences from cattle of different ages revealed that the diversification of germline sequences could be observed in very young calves and that substitution frequency increased with age. The age-dependent accumulation of mutations was particularly apparent in the second IgH complementarity-determining region (CDR2). Single base substitutions were found to predominate, with purines targeted more frequently than pyrimidines and transitions favoured over transversions. In non-coding regions, mutations were detected at a normalised frequency that was indistinguishable from that observed in CDR2. These data are consistent with a process of IgH diversification driven predominantly by somatic hypermutation.     

Neuroprotective effect of naringin by modulation of endogenous biomarkers in streptozotocin induced painful diabetic neuropathy

Diabetes mellitus is a serious debilitating epidemic affecting all social strata in developing as well as developed countries. Diabetic neuropathy is most common of secondary complications associated with diabetes mellitus and is characterized by slowing of nerve conduction velocity, elevated pain, sensory loss and nerve fiber degeneration. The aim of the present investigation was to evaluate the neuroprotective effect of naringin against streptozotocin (STZ) induced diabetic neuropathic pain in laboratory rats. Four weeks after intraperitoneal injection of STZ resulted in significant decrease in mechano-tactile allodynia, mechanical hyperalgesia, thermal hyperalgesia and motor nerve conduction velocity. Activity of endogenous antioxidant like superoxide dismutase as well as membrane bound inorganic phosphate enzyme was also found to be significantly decreased. It not only caused neural cell apoptosis but also enhanced lipid peroxide, nitrite, and inflammatory mediators' (TNF-α) level. Chronic treatment with naringin (40 and 80mg/kg) for 4 weeks significantly and dose dependently attenuated the decrease in level of nociceptive threshold, endogenous antioxidant and membrane bound inorganic phosphate enzyme. It also decreased the elevated levels of oxidative-nitrosative stress, inflammatory mediators as well as apoptosis in neural cells significantly and dose dependently. The important finding of the study is that, the naringin-insulin combination not only attenuated the diabetic condition but also reversed the neuropathic pain, whereas insulin or naringin alone only improved hyperglycemia but partially reversed the pain response in diabetic rats. Thus, naringin is a potential flavonone bearing antioxidant, antiapoptotic and disease modifying property acting via modulation of endogenous biomarker to inhibit diabetes induced neuropathic pain.     

Chirality index, molecular overlay and biological activity of diastereoisomeric mosquito repellents

Both 1-methylisopropyl 2-(2-hydroxyethyl)piperidine-1-carboxylate, (Picaridin((R))) and cyclohex-3-enyl 2-methylpiperidin-1-yl ketone (AI3-37220; 220) have two asymmetric centers, and the four diastereoisomers of each compound are known to have differing degrees of mosquito-repellent activity according to quantitative behavioral assays conducted at the United States Department of Agriculture. Computational chemistry was used to identify the structural and configurational basis for repellent activity. Molecular overlay of the optimized geometries of the lowest energy conformers of the diastereoisomers was investigated to elucidate the role of chiral centers in 220 and Picaridin. It was found that the presence of a chiral carbon alpha to the nitrogen with the S configuration in the piperidine ring is essential to the three-dimensional arrangement of the atoms of the pharmacophore for effective repellent activity.     

Utilisation of sesame (Sesamum indicum) seed meal in broiler chicken diets

1. Soybean meal (SBM) was replaced with sesame seed meal (SSM) at proportions of 0.00, 0.33, 0.67 and 1.00 on a protein basis in broiler diets (2 to 42 d of age). Each diet was fed to 8 replicates of 5 birds each. 2. Body weight gain, food efficiency, ready-to-cook (RTC) yield, relative weights of giblet, liver and abdominal fat were not affected by including SSM up to 0.67 proportion of SBM in starter and finisher diets (360 and 310 g/kg, respectively). The relation between level of SSM and growth performance variable (weight gain, food efficiency and RTC yield) was non-linear. 3. Weight, breaking strength and ash content of tibia decreased non-linearly with the proportion of SSM in diet. 4. The activity of alkaline phosphatase and protein concentration in serum decreased linearly with the amount of SSM in the diet.     

两个玉米自交系苗期叶温的干旱响应研究

利用远红外热成像系统的技术,研究玉米自交系综3与87-1苗期叶片对干旱的响应以及生物量的积累变化。结果表明,在遭受中度干旱胁迫的条件下,87-1的相对生物鲜重(0.716)及相对生物干重(0.799)显著高于综3(0.564,0.658),87-1的叶温差(0.30℃)显著高于综3(0.13℃)。通过离体叶片分析,综3离体叶片较87-1失水速率快,叶温在离体短时间内下降更低。表明在响应水分胁迫方面,87-1较综3叶片蒸发量小,叶温变化大,保水能力好,87-1表现出了更强的耐旱性。生物量积累与叶温差存在紧密的相关性,较高叶温差的基因型在遭受到水分胁迫时生物量积累更强。     

Evaluation of fipronil nanoformulations for effective management of brown plant hopper (Nilaparvata lugens) in rice

In this paper, we report the development, characterization and bioefficacy evaluation of fipronil nanoformulations against brown plant hopper (BPH); Nilaparvata lugens in rice. Nanoformulations were characterized for nanosizing and stability under variable conditions. Bioefficacy of nanoformulations were evaluated under field condition at farms of Indian Agricultural Research Institute, New Delhi during Kharif 2015. At milking stage BPH populations were above economic threshold level; followed by foliar application of fipronil nanoformulations at four doses (viz., 30, 45, 60 and 90 g a.i./ha) along with commercial formulation of fipronil. The mean mortality clearly indicated that all the insecticidal treatments were significantly superior over untreated control where, highest mortalities were observed in nanodispersions followed by monolithic dispersion and commercial formulation of fipronil. Treated plots 7 days after application @ 60 g a.i./ha has shown 93.47%, 86.89% and 80.47% reduction BPH population in nanodispersion, encapsulated monolithic dispersion and commercial formulation treated plots respectively. In comparison to commercial formulation @ 60 g a.i./ha yield has increased by 0.56 t ha^?1 (28.76%) and 0.26 t ha^?1 (14.04%) in nanodispersion and monolithic dispersion.