Comparison of monoclonal and polyclonal antibodies for the detection of canine IgG1 and IgG2, and associations with infection outcome in Leishmania infantum naturally infected dogs

In murine models of leishmaniasis, IgG subclass expression is a proxy measure for Th1/Th2 cellular immune response bias. However, in dogs, the reservoir of zoonotic visceral leishmaniasis, no consistent association has been described between IgG subclass ratios and disease resistance. Inconsistent results may reflect lack of specificity of commonly used commercial antibodies. Our aim was to measure IgG1 and IgG2 responses to crude Leishmania antigen using commercial polyclonal antibodies for comparison with a panel of commercially unavailable monoclonal antibodies, in a cohort of 60 naturally infected dogs, and to compare associations between subclass responses and clinical or parasitological outcomes. IgG1 and IgG2, measured by both antibodies, were higher in clinically symptomatic than in asymptomatic dogs (P ≤ 0.03), reflecting general upregulation of IgG in infected dogs. Unlike the murine model, canine IgG2:IgG1 ratios were not predictive of clinical or parasitological outcomes of infection. Associations between subclass levels and positivity by bone marrow culture and PCR were not consistent when measured with different antibodies. Further research is needed to re-evaluate the specificity of commercially available IgG subclass antibodies.     

Chicken anemia agent in the United States: isolation of the virus and detection of antibody in broiler breeder flocks

Chicken anemia agent (CAA) was isolated from broiler chickens in Texas with a blue wing or anemia dermatitis-like syndrome. Specific-pathogen-free chicks inoculated with field material developed anemia, and CAA was isolated in MDCC-MSB1 cells from bone marrow and lymphoid tissue from inoculated chicks. One isolate, designated EF88/78/276, was further characterized. Infectivity of EF88/78/276 was resistant to treatment with chloroform and with heat at 70 C for 5 minutes. EF88/78/276 was indistinguishable from the Cux-1 and Gifu-1 isolates of CAA by cross-neutralization tests. Almost all 1-day-old susceptible chicks inoculated intramuscularly with EF88/78/276 developed anemia, but contact-infected chicks did not. Antibody to CAA was detected in broiler breeder flocks from Texas, the Delmarva peninsula, and Alabama.     

Production and preliminary characterization of monoclonal antibodies to chicken anemia agent

Mice were immunized with partially purified preparations of the Cux-1 isolate of chicken anemia agent (CAA), and their splenocytes were fused with NSO myeloma cells. Three patterns of staining of CAA-infected cells were recognized when the resulting hybridomas were screened by indirect immunofluorescence (IIF). Hybridomas representative of each staining pattern were cloned, and the monoclonal antibodies (MAbs) were characterized. Type 1 staining was indistinguishable from that produced by polyclonal chicken antisera to CAA. Type 2 staining was confined to large nuclear inclusions. Type 3 staining was predominantly nuclear and granular, and differed from type 1 in being more intense and occurring in a higher proportion of nuclei. Three MAbs producing type 1 staining were predominantly Cux-1-specific by IIF; they also reacted to lower titers with the Gifu-1 isolate but not at all with three other CAA isolates. These MAbs had very slight neutralizing activity against Cux-1. Another MAb giving type 1 staining reacted with all CAA isolates tested to high titers in IIF and neutralization tests. MAbs with type 2 and type 3 staining reacted by IIF with all CAA isolates tested but possessed no neutralizing activity. The availability of MABs to CAA should facilitate development of diagnostic tests for the virus.     

Modelling the impacts of agriculture in mixed-use landscapes: a review and case study involving two species of dabbling ducks

Context

This study synthesizes evidence from three separate surveys of American Black Duck and Mallard breeding habitat usage to quantify the effects of agriculture at the landscape scale.

Objectives

To assess duck breeding activity in agricultural landscapes within the Canadian maritimes in order to measure the overall impact of agricultural land use, the response to particular agricultural activities, and the influence of landscape configuration.

Methods

Models, constructed using a long-term census (SURVEY1), were used to predict habitat selection for two other independent surveys (SURVEY2, SURVEY3). Predictions incorporated information about wetland area and diversity, as well as anthropogenic factors, allowing subsequent analyses to focus on the remaining residual error attributable to agricultural effects.

Results

SURVEY2 results demonstrated that the proportion of active agriculture is an important indicator of the severity of human disturbance, yielding threshold estimates of 39% for Mallards and 60% for Black Ducks, with an overall average of 50%. Agricultural conversion beyond these thresholds deterred breeding ducks independently of other factors. SURVEY3 land cover information demonstrated that the presence of cropland intensified this deterrence effect, even at levels as low as 10%. Woodland cover (in excess of 30%) was important for both species, but its configuration was not.

Conclusions

In addition to quantifying threshold effects, this study reaffirms that woodland is an important part of the maritime landscape matrix, and contributes positively to habitat diversity in mixed use, moderate intensity agricultural regions. Wetland restoration in agricultural landscapes should monitor and promote less crop-intensive, mixed-use practices.

     

Molecular genotyping of multinational ovine and caprine Corynebacterium pseudotuberculosis isolates using pulsed-field gel electrophoresis

Caseous lymphadenitis (CLA) is a chronic, suppurative disease, with a worldwide distribution, caused by Corynebacterium pseudotuberculosis. The clinical manifestation of CLA is known to vary between different countries, and has been postulated to be due to differences in the strains present in these countries. Forty-two sheep and goat isolates of C. pseudotuberculosis from Australia, Canada, Eire, The Netherlands and Northern Ireland were characterized by pulsed-field gel electrophoresis (PFGE), biotyping, antimicrobial susceptibility, and production of phospholipase D. The PFGE-determined genotypes of this multicentric collection were then compared with representative ovine and caprine isolates from a previously published panel of PFGE profiles of United Kingdom isolates. Digestion with SfiI generated 16-18 bands in the 48.5 and 290 kb range, and differentiated four distinct pulsotypes amongst the 36 ovine and 6 caprine strains which displayed remarkable homogeneity. Based on these results, it would appear that the genome of C. pseudotuberculosis is highly conserved, irrespective of the country of strain origin.     

Bovine trypanosomiasis in Southern Tanzania: Parasitological and serological survey of prevalence

In a survey for bovine trypanosomiasis blood smears from 1,617 cattle at 72 sites were examined. Trypanosomes were found in 93 cattle, representing 16% of the cattle in herds in which trypanosomiasis was confirmed. Of the positive cattle 56% had infections with T. congolense, 17% T. vivax and 2.2% T. brucei. Five cattle had mixed infections and in 18 cattle the species was not identified. Sera from 1,352 cattle were tested using microelisa. Ten out of 16 sites, at which no trypanosomes were found in blood smears and at which trypanocides were in use, had over 15% seropositive cattle compared with five of 19 sites at which trypanocides were not in use. It was concluded that the microelisa was a useful aid to the diagnosis of bovine trypanosomiasis and that there is a need for accurate records of drug use and livestock movements to be kept. The serious risk of drug resistant strains of trypanosomes emerging due to the uncontrolled use of trypanocides is emphasised.     

Designing cropping systems for efficient use of limited water in southern Australia

There has been continuous change in the systems of cereal production in southern Australia where the climate is semi-arid with moderately wet winters and hot dry summers. At the outset wheat (c. 1840) was grown in continuous culture as the land was gradually cleared for cropping, but yields declined to uneconomic levels by 1900. That system was then replaced with fallow-wheat rotations, and using phosphorus fertilizer, yields recovered, but not to the original level. From 1945, the high profitability of sheep production encouraged improvement of legume-based pastures and the introduction of pasture-crop sequences – ley-farming. Productivity was greatly increased and the yield of wheat crops regained the original level. But such were the rates of acidification and salinization of soil that reduced productivity and, in the case of salinity, reduced stream quality and threatened adjacent natural ecosystems. This ley-farming system was used until the late 1980s when prices for wool fell dramatically, and farmers reduced sheep numbers and intensified cropping. The thrust since then has been to design cropping systems that are economically as well as environmentally acceptable with salinity control as the major objective. There is now a search for systems that use more water to reduce drainage through the soil and lower saline water tables. A major contribution is the much restricted use of fallow but summer active plants are also required to provide a soil water storage buffer to retain autumn–winter rainfall. New systems under evaluation include herbaceous perennials such as lucerne that are readily integrated into a crop-livestock system and agroforestry combinations with various trees. The design and appropriate distribution of the new systems is aided by remote sensing techniques to locate areas of saline discharge and linkages to zones of groundwater recharge, geographical information systems to arrange the data for land system analysis, and computer models of crops and cropping systems. There is a substantial challenge for farmers, agronomists and others to identify and assess suitable new systems and to bring the public into the debate and into the solution.