Quantitative immunoenzymatic detection of viral encephalopathy and retinopathy virus (betanodavirus) in sea bass Dicentrarchus labrax

Viral encephalopathy and retinopathy disease caused by betanodavirus, genus of the family Nodaviridae, affects marine, wild and farmed species including sea bass, one of the most important farmed species in Europe. This work describes a reliable and sensitive indirect ELISA assay to detect betanodavirus in biological samples using a polyclonal antiserum (pAb 283) against the 283/I09 virus strain, the most common red‐spotted grouper nervous necrosis virus (RGNNV) genotype in the Mediterranean area, and a capture‐based ELISA using a monoclonal antibody (mAb 4C3) specific to a common epitope present on the capsid protein. Using adsorbed, purified VERv preparation, the detection limit of indirect ELISA was 2 μg mL^?1 (3 × 10⁵ TCID50 per mL), whereas for capture‐based ELISA, the sensitivity for the antigen in solution was 17 μg mL^?1 (35 × 10⁵ TCID50 per mL). The capture‐based ELISA was employed to detect VERv in brain homogenates of in vivo infected sea bass and resulted positive in 22 of 32 samples, some of these with a high viral load estimates (about 1.1 × 10⁸ TCID50 per mL). The ELISA system we propose may be helpful in investigations where coupling of viral content in fish tissues with the presence of circulating VERv‐specific IgM is required, or for use in samples where PCR is difficult to perform.     

Establishment and characterization of macrophage cell line from thymus of Catla catla (Hamilton, 1822)

A continuous cell line has been developed from thymus explants of Catla catla and the cells have been subcultured for 63 passages. The cells exhibited optimum growth at 30°C in L‐15 medium containing 15% foetal bovine serum. The cultured cells engulfed yeast cells and fluorescent latex beads. These cells produced reactive oxygen and nitrogen intermediates following stimulation with lipopolysaccharide and phorbol esters. The culture supernatant from the cultured cells had lysozyme activity and these cells demonstrated Fc receptors. Almost all the cells were positive for alpha‐naphthyl acetate esterase enzyme suggesting that the cells are of macrophage lineage and therefore, the cell line was designated as catla thymus macrophage (CTM) cell line. CTM cells formed aggregates around zoospores of Aphanomyces invadans, but were unable to inhibit the germination of spores. The karyotype analysis of CTM cells at 25th passage revealed a typical diploid model with 50 chromosomes per cell. Partial amplification, sequencing and alignment of fragments of two mitochondrial genes 16S rRNA and cytochrome c oxidase subunit 1 confirmed that the CTM cell line originated from C. catla. This cell line should be useful for studying the role of macrophages in differentiation and maturation of thymocytes and can be a source of macrophage‐specific enzymes and cytokines.     

辽宁省棉铃虫虫情划区的调查研究

研究表明,北方特早熟棉区的辽宁省各地棉铃虫的发生程度与气温（年均等温线８～９℃）、降水（年平均降水量５００～７００ｍｍ）及寄主植物的分布三个条件重合程度呈明显的正相关。据此,将本省棉铃虫的发生区域划分为：重发生区、轻发生区、可发生区、偶发生区、不发生区,并绘制出棉铃虫区域划分图。     

Barnyard millet – a potential food and feed crop of future

The two species under genus Echinochloa, E. frumentacea (Indian barnyard millet) and E. esculenta (Japanese barnyard millet), are cultivated for food and fodder by hilly and tribal communities in Asia particularly in India, China and Japan. The crop has wide adaptability and occupies a special place in marginal rainfed areas because of its short life cycle. Although the area under the crop has come down drastically in last 50 years, the crop ability to survive under harsh conditions makes it a better choice during famine years. In the Indian Himalayan region, the crop was traditionally used as a substitute for rice. It has been identified as a suitable choice for climate‐resilient agriculture. High nutrient content and antioxidant effects make it to be considered as a functional food crop. Recently, the demand of the crop has increased due to its highly nutritious grains. Thus, it has the potential to provide both food and nutritional security particularly in hills where nutritional deficiencies are in abundance. Despite enormous potential, the crop has not gained the popularity among masses and is still considered as poor man's food. This work therefore is an attempt to compile the meagre information available on crop history, evolution, crop breeding and present status to make the crop competitive and revamp its cultivation.     

Pyloric outflow obstruction secondary to sclerosing encapsulating peritonitis in a dog

A 6-year-old, male neutered mixed breed dog was presented emergently with a three-week history of hyporexia, vomiting, diarrhoea and weight loss. Upon examination, the patient was dull, had generalised muscle atrophy, moderate abdominal pain and a mild amount of peritoneal effusion. A fluid-filled, distended, corrugated small bowel with marked gastroparesis and moderate peritoneal effusion was noted on abdominal ultrasonography. Endoscopy revealed hyperaemic and friable mucosa and a subjectively narrowed pylorus. Emergency exploratory celiotomy was performed due to worsening patient condition and revealed thick, diffuse, fibrous adhesions of the abdominal cavity. Based on these findings, sclerosing encapsulating peritonitis (SEP) was suspected. A large mass of omentum adjacent to the greater curvature of the stomach had caused a pyloric outflow obstruction. Adhesiolysis was attempted but was unsuccessful due to the friability of the small intestines. The dog was humanely euthanased under anaesthesia. A diagnosis of SEP was confirmed via necropsy. No underlying cause was identified. This is the first known case of a pyloric outflow obstruction secondary to SEP in a dog. Although rare, this condition should be considered as a differential for dogs with signs of a pyloric outflow obstruction with concurrent ascites and abdominal pain, hyporexia, vomiting and diarrhoea.     

Effect of feeding stimulants on the insecticidal properties of Pieris brassicae granulovirus (PbGV) against Pieris brassicae

Feeding stimulants (phagostimulants) increase the food uptake by the host insect larvae and thereby play a vital role in enhancing overall activity of insect viruses formulated as biopesticides. A local strain of granulovirus of Pieris brassicae (PbGV), a serious pest of cole crops worldwide, has shown potential as a biopesticide against this pest and with slight increase in its efficacy can be used successfully as a promising biocontrol agent in integrated pest management (IPM) programs of the pest. The addition of phagostimulants may enhance the efficacy but so far no specific phagostimulant for PbGV has been reported. In the present studies, some reported phagostimulants for insect viruses were tested for their impact on food utilization by host larvae and efficacy of the virus. The specified concentration of virus isolate was mixed with phagostimulants and laboratory studies were conducted to uncover their effect on consumption index (CI) and larval mortality. The studies revealed that phagostimulants, viz. boric acid and crude sugar, when mixed with PbGV, showed highest CI and larval mortality. Consumption index of 1.34, 1.49 and 1.43 mg/mg/day and corresponding larval mortality of 32.9%, 45.8% and 34.2% were recorded with the addition of 0.5% and 1% boric acid and 2% crude sugar, respectively, in PbGV. The leaf extracts of cabbage, cauliflower and broccoli at both the concentrations (1% and 2%) also enhanced the CI but were comparatively less effective than the boric acid and crude sugar. Field evaluation of PbGV alone and in combination with phagostimulants carried out at two geographically isolated locations, viz. Palampur (subtropical) and Sangla (temperate) on three cole crops (cabbage, cauliflower and broccoli) showed that boric acid was the most promising candidate for enhancing the PbGV efficacy under field conditions, followed by crude sugar.