Molecular identification of a begomovirus causing yellow vein disease on Calendula officinalis in India

A whitefly transmitted begomovirus was detected by PCR using begomovirus-specific primers from naturally infected Calendula officinalis plants showing yellow vein disease symptoms. An approximately 800 bp PCR amplicon was cloned and sequenced to identify the species of the virus isolate. Analysis of nucleotide sequence data resulted in its identification as the complete coat protein gene open reading frame (CP ORF) of 771 bp, which encoded 256 amino acid residues. The coat protein of the virus isolate shared maximum identities of 96–97% with four strains of Tobacco curly shoot virus (ToCSV) and an Ageratum enation virus (AgEV) during BLAST analysis of sequence data. Nucleotide- and amino acid-based phylogenetic analysis revealed the close relationship of the isolate with ToCSV strains, therefore it has been identified as an isolate of ToCSV and C. officinalis is considered to be a new host of ToCSV begomovirus. Association of a DNA-β molecule with the virus isolate was also detected by PCR and Southern hybridization tests using DNA-β specific primers and probe.     

Effect of electrical stun/kill method, interval between killing and neck cutting and blood vessels cut on blood loss and meat quality in broilers

1. Broiler chickens were killed using either an electrical waterbath (WB system) delivering 120 mA per bird (50 Hz, alternating current, AC) for 4 s or an alternative stun/kill method (ASK system); where head-only stunning for 1 s was immediately followed by head-to-body (vent) application for 1 s (150 mA, 50 Hz sine wave AC). Within each stun/kill system, the neck was cut ventrally or unilaterally 20, 60, or 180 s after killing. In addition, a control group of broilers was stunned with 100 mA per bird in a waterbath using 1500 Hz AC for 4 s and were bled by a ventral neck cut within 20 s. 2. Blood leaving the neck cut was collected for 90 s in a bin placed on an electronic balance and blood loss (g/kg body weight) calculated. 3. Individually identified, unplucked and uneviscerated carcases were held at ambient temperature until the end of the experimental day and then stored overnight in an air chiller (5 degrees C). The carcases were dissected and the incidence of broken furculum and coracoid bones, haemorrhaging in P. minor and P. major muscles, and discolouration of P. major muscles were determined. 4. When neck cutting was performed in broilers 20 s after the stun or kill, the ASK and WB systems, in comparison with high frequency stunning, produced on average about 10 g per kg less bleed out. Within the stun/kill systems, broilers killed by ASK had a greater bleed out than in the WB system. Neck cutting at 20 s or 60 s post-kill resulted in a greater bleed out than when performed after a delay of 180 s. Ventral or unilateral neck cutting resulted in a similar bleed out. 5. Stunning broilers with 1500 Hz AC resulted in lower incidences of broken bones, haemorrhaging in breast muscles and muscle discolouration post mortem than the stun/kill systems. These defects were significantly lower in the ASK than in the WB system. Delayed neck cutting increased the severity of discolouration occurring Post mortem in the breast muscles. 6. It is suggested that broilers killed by ASK can be neck cut with a delay of up to 180 s without compromising bleed out. The incidence of broken bones and haemorrhaging in breast muscles are significantly less with ASK than WB. 7. Owing to the commercial benefits and potential for improved welfare at slaughter, ASK would appear to be a better method than WB.     

Rocket Immunoelectrophoresis in the Diagnosis of Infectious Bursal Disease

The rocket immunoelectrophoresis (RIE) test was used for the qualitative detection and quantitative estimation of infectious bursal disease virus (IBDV) specific antigen in experimentally infected chickens and samples collected from suspected outbreaks. The IBDV specific antigen was detected in the bursae of experimentally inoculated chickens up to 5 days post infection (PI) by the agar gel precipitation (AGP) test and 7 days PI by the RIE test. The RIE detected IBDV specific antigen in a significantly greater number of samples collected from the field outbreaks than the conventional AGP test. Exudative bursae were found to have a higher antigen content than haemorrhagic bursae and are recommended as the material of choice for diagnosis of IBD. This test could also be used to quantify IBDV specific antigen in commercial killed vaccines.     

Production of recombinant capsid protein of Macrobrachium rosenbergii nodavirus (r‐MCP43) of giant freshwater prawn,M. rosenbergii (de Man) for immunological diagnostic methods

White tail disease (WTD) caused by Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) is a serious problem in prawn hatcheries. The gene for capsid protein of MrNV (MCP43) was cloned into pRSET B expression vector. The MCP43 protein was expressed as a protein with a 6‐histidine tag in Escherichia coli GJ1158 with NaCl induction. This recombinant protein, which was used to raise the antiserum in rabbits, recognized capsid protein in different WTD‐infected post‐larvae and adult prawn. Various immunological methods such as Western blot, dot blot and ELISA techniques were employed to detect MrNV in infected samples using the antiserum raised against recombinant MCP43 of MrNV. The dot blot assay using anti‐rMCP43 was found to be capable of detecting MrNV in WTD‐infected post‐larvae as early as at 24 h post‐infection. The antiserum raised against r‐MCP43 could detect the MrNV in the infected samples at the level of 100 pg of total protein. The capsid protein of MrNV estimated by ELISA using anti‐rMCP43 and pure r‐MCP43 as a standard was found to increase gradually during the course of infection from 24 h p.i. to moribund stage. The results of immunological diagnostic methods employed in this study were compared with that of RT‐PCR to test the efficiency of antiserum raised against r‐MCP43 for the detection of MrNV. The Western blot, dot blot and ELISA detected all MrNV‐positive coded samples as detected by RT‐PCR.     

Assessment of Forest Carbon Stocks in the Himalayas: Does Legal Protection Matter?

Small-scale Forestry - Although protected areas are central to global biodiversity conservation, most land lies within unprotected areas. In the developing world forest habitats have received...     

Influence of age,sex and rearing systems on Toll-like receptor 7 (TLR7) expression pattern in gut,lung and lymphoid tissues of indigenous ducks

1. The objective of the experiment was to determine the influence of age, sex and rearing system on Toll-like receptor 7 (TLR7) gene expression in gut, lung and lymphoid tissues and physiological responses to stress in male and female indigenous ducks of Tamil Nadu, India.

2. A total of 36 ducks (12 males and 24 females) were obtained from local farmers and tissue samples of gut tissues (duodenum, jejunum, ileum and caecum), lymphoid organs (spleen and bursa) and lungs were collected in RNAlater solution followed by RNA extraction.

3. After normalisation to β-actin (endogenous control) qPCR analysis identified a significant effect of age, sex and rearing system on TLR7 expression in the ducks.

4. A significant up-regulation of TLR7 expression was observed in lungs, duodenum, jejunum, ileum and caecum of sexually mature (45 wk) compared with that of immature ducks (16 wk). Among sexes, male ducks had significantly higher TLR7 expression than female ducks.

5. Age and sex interactions were significant in lungs, duodenum, jejunum and caecum. Ducks reared in an extensive housing system showed significantly higher TLR7 expression in bursa, lungs, duodenum, ileum and caecum compared to intensively reared ducks. There were no effects of age, sex and rearing systems on TLR7 expression in the spleen.

6. The heterophil-to-lymphocyte ratio and serum corticosterone were higher in ducks reared on an intensive system compared with ducks from an extensive rearing system.     

Comparison of 4 Direct Coombs' Test Methods with Polyclonal Antiglobulins in Anemic and Nonanemic Dogs for In‐Clinic or Laboratory Use

Background

Difficulties with the direct antiglobulin test (DAT) and its apparent lack of sensitivity and specificity for immune‐mediated hemolytic anemia (IMHA) in dogs have raised skepticism regarding its diagnostic value.

Objective

To compare different DATs and other hematologic parameters in dogs.

Animals

Anticoagulated blood samples from 59 nonanemic and 46 anemic dogs (± IMHA) from a research colony and veterinary clinics.

Methods

Prospective observational study: Immunochromatographic strip, gel microcolumn, and capillary techniques were compared with standard microtiter DAT using 2 polyvalent antiglobulins. Spherocytosis, autoagglutination, osmotic fragility, and clinical data were assessed.

Results

Blood samples from all 59 nonanemic dogs were DAT‐. Among 46 anemic dogs, 33 were suspected of IMHA, but only 20 were DAT+. Old and new DAT methods yielded comparable and consistent results even after storage of chilled blood samples for 1 week. Spherocytosis and autoagglutination (that did not persist after washing) were noted in 15 and 16 DAT+ dogs, respectively. The other 26 anemic dogs, including 21 previously transfused dogs and 4 with autoagglutination, tested DAT‐ by the other methods. Osmotic fragility was increased in 70% (19/27) of anemic and all 15 DAT+ dogs tested. Limited follow‐up testing revealed DAT+ results for 3–70 days.

Conclusions and Clinical Importance

The novel strip and capillary DAT methods are promising adjunct in‐clinic tools. Despite prior immunosuppressive treatment and presence of autoagglutination, the DAT was positive in anemic dogs with IMHA. Transfusion did not cause false DAT+ results. Our results support DAT as a cornerstone in the diagnosis of canine IMHA.     

Compaction of sandy soils for irrigation management

Summary Several irrigation management experiments were conducted at different locations on sandy soils in Haryana State to overcome excessive permeability, poor soil moisture retention and storage in the root zone. Subsurface compaction to 30–40 cm depth created by 6 passes of a 1,500 kg tractor-driven iron roller, 24 to 48 h after irrigation, was found to be beneficial in reducing irrigation requirement. In general, yield of different crops was not affected significantly by surface rolling, except that of mustard which increased significantly. Slight increase in subsurface compaction, about 0.1 g cm^–3, increased the soil moisture retention and reduced the infiltration rate and saturated hydraulic conductivity. Hydraulic conductivity was a better parameter than bulk density for evaluating the effect of rolling. The depth of irrigation water applied in rolling treatment was about 58–74% that of the no-rolling control. Compaction thus reduced water input to these sandy soils without adversely affecting the crop yield.     

Influence of soil inoculation with vesicular-arbuscular mycorrhiza and a phosphate-dissolving bacterium on plant growth and 32P-uptake

The influence of inoculation of soil with a vesicular-arbuscular mycorrhizal fungus (Glomus fasciculatus) and a phosphate-dissolving bacterium (Bacillus circulans) on phosphate solubilization, growth of finger millet (Eleusine coracana) and phosphorus uptake from ³²P-labelled tricalcium phosphate and superphosphate were studied. The mycorrhizal plants produced more dry matter and removed more ³²P from the soil than non-mycorrhizal plants, but did not show increased ³²P activity per unit plant mass. The 30 mm NH₄F-HCl extractable ³²P (available ³²P) in soil, plant ³²P activity and total P uptake were enhanced by soil inoculation with the bacterium. In the treatment receiving both inocula a synergistic effect was recorded with increased P uptake and dry matter production.     

Acidification of Surface Water in Central India

The results of the seasonal analysis of the chemistry of rain, fog and surface water of central India for the two years: June 1996 to May 1998 are presented. A total of 636 rain water samples, 146 fog water samples and 226 surface water samples of 8 locations: namely, Ambikapur, Baikunthpur, Korba, Bilaspur, Raipur, Bhilai, Dallirajhara and Kanker, from different sites were collected for the present study. This paper documents the occurrence of acidic rain and fog events and their effect on surface water pH and aquatic life in central India. Most of the rain and fog water samples collected from Baikunthpur and Korba sites were found to be acidic in nature (i.e. pH < 5). The volume weighted mean pH of rain and fog water samples of these sites was 4.8 and 4.4 with lowest pH 4.4 and 4.0, respectively. However, samples from Ambikapur, Bilaspur, Raipur and Bhilai were slightly acidic and their pH values were always around 5.8. One reservoir (mean pH 5.8) and two stop dams (mean pH 4.4 and 4.2) in Baikunthpur area were found to be acidic. The effect of acidification of water on metal-accumulation in muscles of fishes was studied. Concentrations of Al, Hg, Mn, Cd and Pb were higher in fish from acid, than from less acid waters. Causes of acidification of the atmospheric water and surface water are critically discussed.