Supranational comparison of Neospora caninum seroprevalences in cattle in Germany, The Netherlands, Spain and Sweden.

Herd, within-herd and animal prevalences for Neospora caninum in beef and dairy cattle were compared between four countries. In randomly selected herds from regions of Germany, The Netherlands, Spain and Sweden that were representative for the cattle production of these countries, all animals > or = 2 years were examined serologically by enzyme-linked immunosorbent assays (ELISAs) with high test specificity (> 98.0%). In a previous study, the ELISAs had been validated against each other. Single reacting animals within a herd were confirmed by immunobloting. At the time of sampling, animal (age, breed, herdtype, sex, lactation stage) and herd data (region) were collected. Considerable differences in N. caninum herd, within-herd, and overall animal prevalence estimations were observed between countries, regions, herdtype, age categories and breeds. Herd prevalences, based on confirmation of single reactors, for dairy herds were estimated to be 16% (95%CI: 10-24%) in Sweden, 49% (95%CI: 39-59%) in Germany, 63% (95%CI: 57-69%) in Spain and 76% (95%CI: 67-84%) in The Netherlands and for beef herds 41% (95%CI: 31-50%) in Germany, 46% (95%CI: 41-51%) in Spain and 61% (95%CI: 50-72%) in The Netherlands. No beef herds were examined in Sweden. The lowest animal true prevalence was estimated in dairy cattle in Sweden (0.5% (95%CI: 0.1-0.8%)) while the highest animal true prevalence was estimated for dairy cattle in Spain (16.2% (95%CI: 14.9-17.5%)). Within-herd prevalences varied greatly, with very few farms in Sweden having more than 10% seropositive animals while in Spain more than 10% of the herds had within-herd prevalences between 50 and 100%. Seropositivity was significantly associated with herdtype (beef versus dairy), age, breed and region within countries. The results of this supranational comparative study showed that the importance of N. caninum infection varied greatly within in Europe. Estimates of prevalence can be used to calculate the economic impact of N. caninum infection as well as to evaluate the effect of prevention and control strategies over time.

Comparison and standardisation of serological methods for the diagnosis of Neospora caninum infection in bovines.

Various existing serological tests were compared with a standard panel of 523 sera in a multicentred study across Europe. Well characterised sera from animals that were experimentally or naturally infected with Neospora caninum as well as sera from cattle deemed uninfected with N. caninum were provided by the participants of the study and analysed in several commercial (CHEKIT Dr. Bommeli/Intervet, CIVTEST BOVIS NEOSPORA Hipra, Cypress Diagnostics C.V., Herd Check IDEXX, Mastazyme MAST Diagnostics, P38-ELISA Animal Welfare and Food Safety GmbH (AFOSA)) as well as in-house assays (five ELISAs and one IFAT). Most tests showed a high level of agreement in the interpretation of the test results (positive or negative). A further distinct increase in agreement between tests was obtained after the application of standardised cut-offs offered by a two-graph receiver operating characteristic analysis. This procedure allows a standardised interpretation of results obtained with different tests used in independent, parallel seroepidemiological studies.

An interlaboratory comparison of immunohistochemistry and PCR methods for detection of Neospora caninum in bovine foetal tissues.

Seven European laboratories contributed to a multi-centre evaluation of detection techniques for Neospora caninum in bovine foetuses. Six laboratories participated in immunohistochemistry (IHC) testing. All seven laboratories participated in PCR testing, but the results from one laboratory were not included in the analysis, because of contamination problems in the preparation of the samples. A coded panel of tissue sections from 36 infected and non-infected foetuses was used to evaluate the IHC detection of parasites. A coded panel consisting of 44 homogenized foetal brain samples from natural bovine abortion cases and 32 spiked samples were used to evaluate the PCR methods. Inclusion of a duplicate dilution series of spiked samples was used to evaluate detection limits and repeatability. IHC methods had a relatively low sensitivity, but a high specificity. There was considerable variation in IHC results between participating laboratories, which may be partly explained by examination practices that depended on the experience of the operator. In addition, the use of different antibody reagents, different antibody dilutions, and different enzymatic treatments of tissues may have contributed to the observed variation. PCR methods generally had a higher sensitivity than IHC methods and also a high specificity. The agreement between the majority scores of IHC and PCR methods was low. False positive PCR results indicated contamination problems in some instances. Agreement between the PCR results of the various laboratories was better, compared with the IHC results. There appeared to be no clear relationship between the PCR format (i.e. single or nested) and diagnostic sensitivity. Consequently, an improvement of diagnostic performance of PCR might possibly be achieved by optimizing DNA extraction methods.

Influence of long-term changes in incubation temperature on catecholamine levels in plasma of chicken embryos (Gallus gallus f. domestica).

Catecholamine concentrations were determined from day 18 to 21 of incubation (D18, D21) in developing chicken embryos. The control group was continuously incubated at 37.5 degrees C. The eggs of the two other groups were incubated at 37.5 degrees C until day 14. In the cold group, temperature was decreased to 35.0 degrees C and in the warm group, incubation temperature was increased to 38.5 degrees C for the remainder of incubation. Plasma catecholamine concentrations were measured in eggs exposed to a change in incubation temperature for 4, 5, 6 and 7 days. Embryos in the warm group had dopamine (DA) and noradrenaline (NA) concentrations that were significantly higher than in the control group. On the contrary, eggs incubated at the cooler temperature had hormone levels that were significantly lower than in the control group. Adrenaline (A) levels in the two experimental treatments were significantly lower compared to control eggs. Temperature modulated the time needed for development. Chicken embryos are supposed to hatch on day 21. However, on day 20, NA concentration in the cold-incubated group was too low to fulfill its essential physiological function, whereas in the warm group, the NA concentration seems to be sufficient. Long-term exposure to altered incubation temperature affects the quantitative catecholamine concentration during development, but the relative proportion of each catecholamine remained constant.