A PCR approach (transposon PCR) with primers based on repetitive transposon-like sequences, whichdepending on the isolatewere found at a minimum frequency of 19 on the C. burnetii genome, was established for the highly sensitive and specific detection of C. burnetii. This study describes the analytical detection of C. burnetii in milk, which requires a special preparation method prior to PCR. Because of the low level of C. burnetii particles in milk samples, template DNA was concentrated by a factor of 200, using cetyltrimethylammonium bromide as the precipitation reagent. Using this particular preparation method, even a single C. burnetii particle could be detected in 1 ml milk.

BackgroundAvian trichomonosis is known as a widespread disease in columbids and passerines, and recent findings have highlighted the pathogenic character of some lineages found in wild birds. Trichomonosis can affect wild bird populations including endangered species, as has been shown for Mauritian pink pigeons Nesoenas mayeri in Mauritius and suggested for European turtle doves Streptopelia turtur in the UK. However, the disease trichomonosis is caused only by pathogenic lineages of the parasite Trichomonas gallinae. Therefore, understanding the prevalence and distribution of both potentially pathogenic and non-pathogenic T. gallinae lineages in turtle doves and other columbids across Europe is relevant to estimate the potential impact of the disease on a continental scale.ResultsWe examined 281 samples from four wild columbid species for Trichomonas infection and determined the genetic lineages. The overall prevalence was 74%. There were significant differences between the species (P = 0.007). The highest prevalence was found in stock doves Columba oenas (86%, n = 79) followed by wood pigeons Columba palumbus (70%, n = 61) and turtle doves (67%, n = 65), while three of five collared doves Streptopelia decaocto (60%) were infected. We found seven different lineages, including four lineages present in columbids in the UK, one lineage already described from Spain and three new lineages, one of those found in a single turtle dove migrating through Italy and another one found in a breeding stock dove. Stock doves from Germany and collared doves from Malta were infected with a potentially pathogenic lineage (lineage A/B), which is known to cause lesions and mortality in columbids, raptors and finches.ConclusionsGenerally, turtle doves showed high prevalence of Trichomonas infection. Furthermore, the potentially pathogenic lineage A/B (or genotype B according to previous literature) was found in a recovering stock dove population. Both findings are worrying for these columbid species due to the occasional epidemic character of trichomonosis, which can have severe negative effects on populations.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-2170-0) contains supplementary material, which is available to authorized users.

A "nested" PCR approach with primers based on conserved plasmid sequences was used for the highly sensitive and specific detection of Coxiella (C.) burnetii in clinical samples collected from cattle, dogs, cats and humans. Results were in good agreement with those obtained from Capture-ELISA and isolation of the organism in BGM cell culture. We also tested primers with sequences derived from genomic DNA and sequences based on 16S rRNA. In addition, we applied PCR for the differentiation of C. burnetii plasmid types from 28 isolates originating from the USA, Europe and South Africa. Reference isolates Nine Mile RSA493, Dugway 5J108-111 and all European isolates tested were recognized only by primers specific for the QpH1 plasmid. One isolate from a goat abortion in Namibia reacted identically to the reference isolate Priscilla Q177 bearing the QpRS plasmid. Reference isolate S Q217 with plasmid sequences integrated into the genome reacted with none of the plasmid-specific primer pairs.

15The Porcine Reproductive and Respiratory Syndrome (PRRS) is one of the economically 16 most important swine diseases worldwide. The virus can be spread by viraemic and 17 persistent infected pigs. Spread and infection are advanced in areas with high herd and 18 population densities. As wild boars have been found seropositive for PRRSV in 19Germany, France, and the USA, exchange between wild and domestic pig populations 20 may exist. However, comprehensive information on PRRSV infection in wild boars is 21 presently not available. The aim of the current study was thus, to systematically study 22 PRRSV infection in wild boars to provide information on spatiotemporal, host and viral 23 effects. The study was based on 531 wild boars from 52 hunts in Germany (2004Germany ( -2007. 24 PRRSV-infection was determined and strains (US/EU) were classified by PCR. A total of 2515.9% of the wild boars were PRRSV-positive (US: 14.2%; EU: 6.2%), with remarkable 26 effects of state (US: 5.1 -46.2%; EU: 0 -17.6%), season (0 -36.5%) and tissue (lungs: 27 89%; tonsils: 11%). Prevalences did neither correlate with age or weight, nor with density 28 of production units, domestic pigs or wild boars. ORF 1-sequences within EU and US 29 strains did not differ among wild boar samples. Homologies between EU-30 samples/Lelystad-Virus and US-samples/PRRSV-MLV virus were 99. 3% and 97%, 31 respectively. This is the first comprehensive evidence of PRRSV infection in wild boars. 32We conclude that there is only a weak relation between wild boar and domestic PRRSV 33 infection. 34 35