Production and evaluation of monoclonal antibodies for the detection of beet mild yellowing luteovirus and related strains

A sugar-beet-infecting isolate of beet mild yellowing luteovirus (BMYV), and aBrassica-infecting isolate of beet western yellows luteovirus (BWYV) were used to produce monoclonal antibodies for epidemiological studies with BMYV and related field strains. Thirty-four monoclonal antibodies were tested for their reaction with 9 luteoviruses in triple-antibody-sandwich enzyme-linked immunosorbent assay. One (MAFF 24) is now routinely used in the UK for detecting BMYV and BWYV in plants and aphids, although it does not discriminate between them. Heterologous reactions were detected between some of the monoclonal antibodies and potato leafroll virus (PLRV), bean leafroll virus (BLRV) and barley yellow dwarf virus (BYDV-RPV). 38% of antibodies raised to BWYV reacted with PLRV compared with 4% of those raised to BMYV. Monoclonal antibodies were produced which distinguished a sugar-beet-infecting isolate of BMYV with differing host range and serological properties from the commonly-occurring field strain.     

A comparative study of carbofuran metabolism in treated and untreated soils

Soils which have been pretreated with carbofuran can degrade the insecticide more rapidly than untreated soils, with a consequent loss of efficacy. In laboratory studies, soils pretreated with carbofuran were found to degrade the chemical more rapidly than soils which were not so pretreated. When pretreated soils were sterilised, the rate of carbofuran degradation was much reduced, indicating that most of it was due to microbial action. Incubation of pretreated soil with [phenyl-U-¹⁴C]carbofuran led to the rapid disappearance of the parent compound (3 % left after seven days). Most of the ¹⁴C was accounted for as bound residue after seven days, whilst smaller amounts were recovered as carbon dioxide, 3-hydroxycarbofuran, 3-ketocarbofuran, and an unknown metabolite. Incubation of pretreated soil with [carbonyl-¹⁴C]carbofuran led to rapid loss of the parent compound and the recovery of 73% of ¹⁴C as carbon dioxide by five days. Most of the bound ¹⁴C (>90%) arising from [phenyl-U-¹⁴C]carbofuran treatment of pretreated soil was extracted by 1 M sodium hydroxide and about half of the extracted ¹⁴C was precipitated with ‘humic acids’ after acidification. These and other results suggest that the major metabolic route for carbofuran in pretreated soils involves hydrolysis of the ester bond leading to (1) release of carbofuran phenol which rapidly binds to soil organic matter and, (2) release of the carbonyl moiety which quickly degrades to generate carbon dioxide.     

The immune response and PBMC subsets in canine visceral leishmaniasis before, and after, chemotherapy

Peripheral blood mononuclear cell subsets, in vitro lymphoproliferative response to leishmanial antigen, and Leishmania-specific serum antibody levels were examined in 11 dogs, naturally infected with L. infantum, and 9 healthy control dogs. A decrease in the percentage of CD4+ T-cells and an increase in the proportion of gammadelta T-cells and sIgG+ B-cells were observed during canine visceral leishmaniasis (CVL). These changes may be responsible for the marked humoral response and the absence of in vitro lymphoproliferation to mitogen and specific parasite antigens. This possibility was supported by the analysis of these subsets after treatment with amphotericin B. One month after therapy, a significant increase in the percentage of CD4+ T-cells and a decrease of gammadelta T-cells and sIgG+ B-cells were observed. At the same time, the lymphocyte blastogenesis assay with leishmanial antigen was positive and the levels of specific antibodies to Leishmania were significantly lower than before the treatment. Five months after therapy, lymphocyte proliferative response to LSA disappeared, antibody and lymphocyte subsets levels returned to those observed during CVL. Therapeutic failure in CVL is associated with the inability of antileishmanial drugs to completely revert the profound immunodepression induced by the infection and prevent relapse.     

Sensitivity analyses for four pesticide leaching models

Sensitivity analyses using a one-at-a-time approach were carried out for leaching models which have been widely used for pesticide registration in Europe (PELMO, PRZM, PESTLA and MACRO). Four scenarios were considered for simulation of the leaching of two theoretical pesticides in a sandy loam and a clay loam soil, each with a broad distribution across Europe. Input parameters were varied within bounds reflecting their uncertainty and the influence of these variations on model predictions was investigated for accumulated percolation at 1-m depth and pesticide loading in leachate. Predictions for the base-case scenarios differed between chromatographic models and the preferential flow model MACRO for which large but transient pesticide losses were predicted in the clay loam. Volumes of percolated water predicted by the four models were affected by a small number of input parameters and to a small extent only, suggesting that meteorological variables will be the main drivers of water balance predictions. In contrast to percolation, predictions for pesticide loss were found to be sensitive to a large number of input parameters and to a much greater extent. Parameters which had the largest influence on the prediction of pesticide loss were generally those related to chemical sorption (Freundlich exponent nf and distribution coefficient Kf) and degradation (either degradation rates or DT50, QTEN value). Nevertheless, a significant influence of soil properties (field capacity, bulk density or parameters defining the boundary between flow domains in MACRO) was also noted in at least one scenario for all models. Large sensitivities were reported for all models, especially PELMO and PRZM, and sensitivity was greater where only limited leaching was simulated. Uncertainty should be addressed in risk assessment procedures for crop-protection products.     

A decision-support system for Salmonella in broiler-chicken flocks

We built a decision-support system to assess the risk of contamination of chicken-broiler flocks by Salmonella at the end of the rearing period. This system was developed from the survey data from 85 chicken-broiler flocks located in western France. First, we estimated the probability of contamination of the house by Salmonella before placement of day-old chicks via a cleansing inspection using a visual-inspection grid, a decontamination evaluation using count-plates, and risk factors for Salmonella persistence in the barn after cleansing and disinfection. Second, we estimated (using a logistic model) the probability of prevalent contamination of the flock by Salmonella at the end of the rearing period. Validation was carried out on 60 flocks selected from seven production companies in western France. The risk estimated by the model was compared to the Salmonella status of the flock (gold standard) assessed by samples taken from the environment of the broilers and analysed with classical bacteriological methods. The sensitivity was 97.8% and the specificity 64.3%.     

Differences in metabolic parameters and gene expression related to osteochondrosis/osteoarthrosis in pigs fed 25-hydroxyvitamin D3

Osteochondrosis/osteoarthrosis (OC/OA) are common terms for various joint pathologies that occur in pigs. Pathologies that may contribute to these disorders have been described, but the primary cause(s) remain unknown. We hypothesised that as OC has some similarities to dyschondroplasia, which involves a failure of growth plate chondrocytes to fully differentiate and hypertrophy, treatment with 25-hydroxyvitamin D3 (25-D) might reduce the incidence and/or severity of lesions in pigs, as it does in chickens with dyschondroplasia. Control pigs were fed a commercial diet ad libitum. In the treated group this diet was supplemented with 25-D at 0.1 mg/kg. Ten pigs from each of the control and treated groups were sampled at 7, 12, 16 and 21 weeks. Treatment with 25-D had no effect on the incidence or severity of OC/OA lesions. Cartilage dry weight, total collagen content and proteoglycan content, and plasma levels oftotal calcium, inorganic phosphorous, vitamin C, insuline-like growth factor-I, parathyroid hormone and tumour necrosis factor alpha were unaffected by treatment. In addition, none of these parameters were correlated with the incidence or severity of OC/OA lesions. The mRNA expression levels of 21 out of 23 genes assayed by RT-PCR were unaltered in articular cartilage from OA lesion samples as compared to normal articular cartilage. However, collagen type II was reduced and collagen type X increased in OA lesion and near lesion samples. These results suggest that OA in pigs may share some features of osteoarthritis in other mammalian species.     

Quantitative analysis of production traits in saltwater crocodiles (Crocodylus porosus): III. juvenile survival

Mortality records of 1302 juvenile crocodiles were available for analysis. Crocodiles that were slaughtered during this study were treated as censored (n = 2151). Additionally, records from animals that had neither died nor been slaughtered, i.e. were still alive in the production system (n = 1582), were censored at the last date of data collection. There were a total of 3733 censored records. The data were all full‐sib records from 29 parental pairs from Janamba Croc Farm (Northern Territory, Australia), collected over nine consecutive years. Data were analysed using an extension of Cox's proportional hazards model to include frailty (random) terms to account for genetic effects. Heritability of log survival time for juvenile crocodile survival was 0.15 (SE 0.04). The probability of a juvenile crocodile surviving to day 400 was estimated to be only 51%. These results are the first to quantify juvenile survival in a captive breeding situation. Also, this is the first heritability estimate of crocodile survival and is a fundamental element in the development of a genetic improvement programme.