Comparison of the complement fixation test and competitive ELISA for serodiagnosis of Anaplasma marginale infection in experimentally infected steers

OBJECTIVE: To compare sensitivity of a complement fixation (CF) test and competitive ELISA (cELISA) for detection of Anaplasma marginale in experimentally infected steers. ANIMALS: 40 crossbred (Angus-Simmental) steers. PROCEDURES: Steers were inoculated with 2.6 x 10(9) A marginale-infected erythrocytes (day 0). Blood samples were collected on days 9, 13, 20, 28, 34, 41, 61, 96, 126, and 156 days after inoculation. The percentage of parasitized erythrocytes (PPE) was determined by microscopic examination of stained blood films, and sera were evaluated with the CF test and cELISA by use of USDA-approved methods. Sensitivity and agreement (kappa statistic) between the 2 methods were determined. Persistent infections were confirmed by inoculation of blood obtained from infected steers into susceptible, splenectomized calves. RESULTS: 9 days after inoculation, sensitivity of the cELISA was 47.5%, whereas the CF test failed to identify seropositive steers. After day 13, sensitivity of the cELISA and CF test was 100% and 20%, respectively. During peak parasitemia (day 20), sensitivity of the cELISA and CF test was 100%. Thereafter, sensitivity of the CF test fluctuated between 7.5% and 37.5%, whereas sensitivity of the cELISA remained at 100%. Overall sensitivity of the cELISA and CF test was 94.8% and 26.5%, respectively (kappa statistic, 0.039). CONCLUSIONS AND CLINICAL RELEVANCE: The cELISA had superior sensitivity for serologic detection of A marginale.The CF test and cELISA each had a high percentage of false-negative results during the prepatent period. These findings are relevant for export certification and anaplasmosis prevention or eradication programs.     

Shiga toxin-producing Escherichia coli and their bacteriophages as a model for the analysis of virulence and stress response of a food-borne pathogen

Enterohemorrhagic Escherichia coli (EHEC) are a subgroup of Shiga toxin-producing Escherichia coli (STEC) that are able to cause serious food-borne intestinal diseases which can be followed in 5 to 15% by extraintestinal sequelae such as the hemolytic-uremic syndrome (HUS). One of the major pathogenicity factors of EHEC is the production of one or more Shiga toxins (Stx), which act as inhibitors of protein biosynthesis and have profound effects on the signal transduction and immunological response in eukaryotic cells. The stx genes are located in the genome of heterogeneous, lambdoid, functional or cryptic bacteriophages and are expressed during the phage life cycle. Due to the linkage between the phage life cycle and stx expression, STEC and their bacteriophages are useful as a model for the analysis of stress response and virulence of this food-borne pathogen.     

Expression of porcine endogenous retroviruses (PERVs) in melanomas of Munich miniature swine (MMS) Troll

Porcine endogenous retroviruses (PERVs) are integrated in the genome of all pig breeds. Since some of them are able to infect human cells, they might represent a risk for xenotransplantation using pig cells or organs. However, the expression and biological role of PERVs in healthy pigs as well as in porcine tumours is largely unknown. Since we and others have recently shown overexpression of a human endogenous retrovirus, HERV-K, in human melanomas, we studied the expression of PERVs in melanomas of selectively bred Munich miniature swine (MMS) Troll. This breeding herd of MMS Troll is characterised by a high prevalence of melanomas, which histologically resemble various types of cutaneous melanomas in humans. Several genetic factors have been defined when studying inheritance of melanomas and melanocytic nevi in MMS Troll. Here we show that the polytropic PERV-A and PERV-B as well as the ecotropic PERV-C are present in the genome of all melanoma bearing MMS Troll investigated. Most interestingly, in the spleen, but not in other organs, recombinant PERV-A/C proviruses were found. PERV expression was found elevated in melanomas when compared to normal skin and viral proteins were expressed in melanomas and pulmonary metastasis-derived melanoma cell cultures. During passaging of these cells in vitro the expression of PERV mRNA and protein increased and virus particles were released as shown by RT activity in the supernatant and by electron microscopy. Genomic RNA of PERV-A, -B and -C were found in pelleted virus particles. Although PERV expression was elevated in melanomas and pulmonary metastasis-derived cell cultures, the function of the virus in tumour development is still unclear.     

Contrasting edge and pasture matrix effects on ant diversity from fragmented landscapes across multiple spatial scales

Landscape Ecology - Harmful effects of habitat loss and fragmentation can be detected across multiple spatial scales, yet most studies that aim to characterize these effects take place at a single...     

Land-use history has a stronger impact on soil microbial community composition than aboveground vegetation and soil properties

The response of soil microbial communities following changes in land-use is governed by multiple factors. The objectives of this study were to investigate (i) whether soil microbial communities track the changes in aboveground vegetation during succession; and (ii) whether microbial communities return to their native state over time. Two successional gradients with different vegetation were studied at the W. K. Kellogg Biological Station, Michigan. The first gradient comprised a conventionally tilled cropland (CT), mid-succession forest (SF) abandoned from cultivation prior to 1951, and native deciduous forest (DF). The second gradient comprised the CT cropland, early-succession grassland (ES) restored in 1989, and long-term mowed grassland (MG). With succession, the total microbial PLFAs and soil microbial biomass C consistently increased in both gradients. While bacterial rRNA gene diversity remained unchanged, the abundance and composition of many bacterial phyla changed significantly. Moreover, microbial communities in the relatively pristine DF and MG soils were very similar despite major differences in soil properties and vegetation. After >50 years of succession, and despite different vegetation, microbial communities in SF were more similar to those in mature DF than in CT. In contrast, even after 17 years of succession, microbial communities in ES were more similar to CT than endpoint MG despite very different vegetation between CT and ES. This result suggested a lasting impact of cultivation history on the soil microbial community. With conversion of deciduous to conifer forest (CF), there was a significant change in multiple soil properties that correlated with changes in microbial biomass, rRNA gene diversity and community composition. In conclusion, history of land-use was a stronger determinant of the composition of microbial communities than vegetation and soil properties. Further, microbial communities in disturbed soils apparently return to their native state with time.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.     

Cover crop and plant density in seed production of white clover (Trifolium repens L.)

Seed crops of white clover (Trifolium repens L.) are usually established with a cover crop. Provided sufficient light, white clover may compensate for low plant density by stoloniferous growth. Our objectives were (1) to compare spring barley or spring wheat used as cover crops for white clover and (2) to find the optimal seeding rate/row distance for white clover. Seven field trials were conducted in Southeast Norway from 2000 to 2003. Barley was seeded at 360 and 240 seeds m^?2 and wheat at 525 and 350 seeds m^?2. White clover was seeded perpendicularly to the cover crop at 400 seeds m^?2/13 cm row distance or 200 seeds m^?2/26 cm. Results showed that light penetration in spring and early summer was better in wheat than in barley. On average for seven trials, this resulted in 11% higher seed yield after establishment in wheat than in barley. The 33% reduction in cover crop seeding rate had no effect on white clover seed yield for any of the cover crops. Reducing the seeding rate/doubling the row distance of white clover had no effect on seed yield but resulted in slightly earlier maturation of the seed crop.     

Constructed wetlands as a treatment method for effluents from intensive trout farms

This study examined the effects of different hydraulic loading rates on the treatment efficiency of subsurface flow (SSF) constructed wetlands treating effluents from trout farming over a period of 6 months. Six identical wetland cells with a pre-sedimentation zone of 9.6 m² and a root zone of 23.6 m² were loaded with effluents from intensive trout farming (> 2.1 kg feeding stuff per L/s and day). The total runoff of 13.2 L/s was treated in the wetland cells, where two duplicate cells received equal hydraulic loads of 3.9, 1.8 and 0.9 L/s. All examined wetland cells had significant treatment effects on the nutrient fractions containing particulate matter [total nitrogen (TN), total phosphorous (TP), biological oxygen demand in 5 days (BOD₅), chemical oxygen demand (COD), and total suspended solids (TSS)].

Efficiency was between 5.5% for TN and 90.1% for TSS. The SSF wetland also had a high treatment effect on total ammonia nitrogen (TAN), with efficiencies of 61.2 to 87.8%. Nitrate nitrogen (NO₃–N) and phosphate phosphorous (PO₄–P) showed a significant increase in the wetland effluent by 8.4 to 209%. Nitrite nitrogen (NO₂–N), had no significant, or significant effluent increase depending on the inflow rate. Treatment efficiency for particulate nutrients and TAN increased with decreasing hydraulic load, while the differences between 1.8 and 0.9 L/s were not significant. The treatment efficiency for TP was constant for all cells, at around 40%. The wetland receiving 3.9 L/s was over-flooded after 10 to 12 weeks due to colmatation. Nevertheless, the wetland still showed high treatment efficiencies. For commercial trout farms, SSF wetlands are a highly effective method of effluent treatment. A hydraulic load of 1 L/s on 13.3 m² wetland area (1.8 L/s on the examined wetland) seems most suitable. Higher loads lead to accelerated wetland colmatation, while lower loads waste space.