The use of a geographic information system to identify a dairy goat farm as the most likely source of an urban Q-fever outbreak.

BACKGROUND: A Q-fever outbreak occurred in an urban area in the south of the Netherlands in May 2008. The distribution and timing of cases suggested a common source. We studied the spatial relationship between the residence locations of human cases and nearby small ruminant farms, of which one dairy goat farm had experienced abortions due to Q-fever since mid April 2008. A generic geographic information system (GIS) was used to develop a method for source detection in the still evolving major epidemic of Q-fever in the Netherlands. METHODS: All notified Q-fever cases in the area were interviewed. Postal codes of cases and of small ruminant farms (size >40 animals) located within 5 kilometres of the cluster area were geo-referenced as point locations in a GIS-model. For each farm, attack rates and relative risks were calculated for 5 concentric zones adding 1 kilometre at a time, using the 5-10 kilometres zone as reference. These data were linked to the results of veterinary investigations. RESULTS: Persons living within 2 kilometres of an affected dairy goat farm (>400 animals) had a much higher risk for Q-fever than those living more than 5 kilometres away (Relative risk 31.1 [95% CI 16.4-59.1]). CONCLUSIONS: The study supported the hypothesis that a single dairy goat farm was the source of the human outbreak. GIS-based attack rate analysis is a promising tool for source detection in outbreaks of human Q-fever.

[Reintroduction of E. granulosus by import of cows in the Netherlands]. / Herintroductie van echinococcose via runderimport in Nederland.

Since East European countries joined the EU, the import of both dairy and beef cows from these countries increased considerably. Based on the identification and registration system it turned out that in the period from May until December 2007 about 200 cows per month were imported from Romania. These animals were either slaughtered immediately or in autumn. In autumn, cysts were noticed both in slaughtered cows during meat inspection and in deceased animals (originated from Romania) during postmortem investigation performed by the Animal Health Service. Because cysts were strongly reminiscent of Echinococcus granulosus hydatid cysts, samples were sent to the authorized laboratory (National Reference Laboratory of Parasitology), where the reintroduction of this potentially zoonotic parasitic infection has been confirmed. The risks of reintroduction of E. granulosus in the Netherlands are described.

Low predictive value of seroprevalence of Toxoplasma gondii in cattle for detection of parasite DNA.

The role of beef in human infections with Toxoplasma gondii is not clear. To get a better understanding of the value of seroprevalence as an indication of the role of beef in human infections with T. gondii we studied the seroprevalence of T. gondii in Dutch cattle and analysed the correlation between detection of antibodies and parasitic DNA. An indirect ELISA was developed and used to test a sample of the Dutch cattle population. Since validation of the ELISA was hampered by a lack of sufficient bovine reference sera, the results were analysed in two different ways: using a cut-off value that was based on the course of the OD in 27 calves followed from birth until 16 months of age, and by fitting a mixture of two normal distributions (binormal mixture model) to the log-transformed ODs observed for the different groups of cattle in the study population. Using the cut-off value, the seroprevalence was estimated at 0.5% for white veal, 6.4% for rosé veal and 25.0% for cattle. However, using the frequency distributions the prevalences were higher: 1.9% for white veal, 15.6% for rosé veal and 54.5% for cattle. Next, for 100 cattle the results with two different serological assays (ELISA and Toxo-Screen DA) were compared with detection of parasites by our recently developed sensitive magnetic capture PCR. Toxoplasma gondii DNA was detected in only two seronegative cattle. This discordance demonstrates that seroprevalence cannot be used as an indicator of the number of cattle carrying infectious parasites. Demonstrating parasitic DNA in seronegative cattle and not in seropositive cattle suggests that only recent infections are detectable. Whether beef from these PCR-positive cattle is infectious to humans remains to be studied.

Evaluation of ELISA test characteristics and estimation of Toxoplasma gondii seroprevalence in Dutch sheep using mixture models.

Lamb and mutton are considered important sources of human Toxoplasma gondii infections, but actual data on the prevalence of T. gondii in sheep in The Netherlands is lacking. The aim of this study was to investigate the prevalence of T. gondii in slaughtered sheep to get more insight in the importance of sheep as a source of human infection. In addition, regional variation in prevalence was studied, as this may indicate differences in environmental contamination. An in-house ELISA that detects antibodies against T. gondii was developed and used to test 1179 sera collected from sheep presented at 11 Dutch slaughterhouses between October and December 2007. Since validation of the serological assay was hampered by a lack of appropriate reference sera, the diagnostic performance and seroprevalence were estimated by fitting a binormal mixture model. ROC-curve analysis on the fitted distributions showed high discriminatory power (AUC=0.995), and high sensitivity and specificity of the ELISA. The overall prevalence was estimated at 27.8% (25.6-29.9%), but was significantly higher in sheep over 1 year old, and in sheep from the central provinces. The high sensitivity and specificity of the in-house ELISA were confirmed by Bayesian analysis together with three commercially available assays: Toxo-Screen DA (bioMérieux), Chekit Toxotest Antibody ELISA (IDEXX), and Toxoplasmosis serum screening ELISA (Institut Pourquier). In conclusion, the binormal mixture model proved a useful method to obtain estimates of diagnostic performance and seroprevalence without use of reference sera. The seroprevalence in sheep was high, and as sheep with antibodies usually carry tissue cysts, this indicates that undercooked lamb and mutton may indeed be important sources of human toxoplasmosis in The Netherlands.