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.     

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.     

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.     

Fate of [14C]daminozide and [14C]maleic hydrazide in American elm (Ulmus americana L.)

Radiolabelled daminozide and maleic hydrazide (MH) were injected into American elm seedlings, kept in nutrient solution, to determine their translocation pattern and metabolic fate. Both compounds were rapidly translocated to all parts of the plant. After 21 days, 13% of the applied ¹⁴C was exuded into the nutrient solution from the roots of the plants treated with MH. Using gel-filtration and thin-layer chromatographic techniques, it was determined that daminozide did not form any metabolite, and that MH was converted into a MH-sugar complex. A significant amount of ¹⁴C was unextractable from the plant tissue.     

Adult‐onset,chronic, cyclic thrombocytopenia in a Rhesus macaque (Macaca mulatta) after dengue virus vaccination and viral challenge

An 8‐year‐old, male Rhesus macaque (Macaca mulatta), previously used for dengue virus (DENV) vaccine research with viral challenge, was presented with adult‐onset, chronic, cyclic thrombocytopenia. Platelet number, morphology, and function were evaluated by automated hematology, peripheral blood smears, electron microscopy, flow cytometry, and impedance aggregometry. Bone marrow was evaluated by cytology. Both serum anti‐dengue nonstructural protein 1 (NS1) antibodies and anti‐platelet antibodies were detected by ELISA. Platelet characterization showed a lack of aggregation to all agonists (ADP, ASP, and collagen), increased activation with increased expression of surface marker (HLA‐ABC), and an absence of surface receptor GPIX during clinical episodes of petechiae and ecchymoses, even in the presence of normal platelet counts. Bone marrow aspirates identified potential mild megakaryocytic hypoplasia. All platelet functions and morphologic attributes were within normal limits during clinically normal phases. Presence of anti‐dengue NS1 serum antibodies confirmed a positive DENV titer 8 years postvaccination. Based on the history and clinical findings, a primary differential diagnosis for this chronic, cyclic platelet pathology was autoimmune platelet destruction with potential bone marrow involvement.     

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。     

Infectious dose‐dependent accumulation of live highly pathogenic avian influenza H5N1 virus in chicken skeletal muscle—implications for public health

Highly pathogenic avian influenza viruses (HPAIV) of H5N1 subtype are a major global threat to poultry and public health. Export of poultry products, such as chicken and duck meat, is a known source for the cross‐boundary spread of HPAI H5N1 viruses. Humans get infected with HPAI H5N1 viruses either by close contact with infected poultry or through consumption of fresh/undercooked poultry meat. Skeletal muscle is the largest soft tissue in chicken that has been shown to contain virus during systemic HPAIV infection and supports productive virus infection. However, the time between infection of a chicken with H5N1 virus and presence of virus in muscle tissue is not yet known. Further, it is also not clear whether chicken infected with low doses of H5N1 virus that cause non‐fatal subclinical infections continue to accumulate virus in skeletal muscle. We investigated the amount and duration of virus detection in skeletal muscle of chicken experimentally infected with different doses (10², 10³ and 10⁴ EID₅₀) of a HPAI H5N1 virus. Influenza viral antigen could be detected as early as 6 hr after infection and live virus was recovered from 48 hr after infection. Notably, chicken infected with lower levels of HPAI H5N1 virus (i.e., 10² EID₅₀) did not die acutely, but continued to accumulate high levels of H5N1 virus in skeletal muscle until 6 days post‐infection. Our data suggest that there is a potential risk of human exposure to H5N1 virus through meat from clinically healthy chicken infected with a low dose of virus. Our results highlight the need to implement rigorous monitoring systems to screen poultry meat from H5N1 endemic countries to limit the global spread of H5N1 viruses.     

Effect of Castor Oil on Soil Erosion by Water

Soils from six different regions of the State of Uttar Pradesh, India, were treated with 1% and 2% concentrations of castor oil and incubated for 15 days and 30 days time. The erosion coefficients of natural as well as treated soils were determined with a simple laboratory erosion apparatus. The castor oil was found to be highly effective in reducing soil erosion. It was more effective in light texture soils with reduction in erosion coefficients to the extent of 88 70. Heavy textured soils also registered good reduction in erosion coefficient, to the extent of 20 %. The castor oil was effective in both the concentrations and both the incubation periods. Similarly, it was effective at all the four slope levels viz. 1, 2, 4 and 8%. The castor oil has been found much superior than cellulose and protein as far as their capability to check soil erosion is concerned.