Using the flagging method, questing ticks were collected at four sites in 2010-2012 and 2000 D. reticulatus were analysed regarding infection with Rickettsia, Babesia, Borrelia and Anaplasmataceae by conventional or real-time PCR.  PCR analyses showed an average prevalence of 64% for Rickettsia sp. 
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Dermacentor reticulatus in Berlin/Brandenburg (Germany): Activity patterns and associated pathogens.Dermacentor reticulatus is one of the most important European tick species.  However, its spatial distribution, seasonality and regional vector role are not well known.  This study aimed to gather information about abundance patterns of questing ticks and associated pathogens in unfed female adult D. reticulatus in the Berlin/Brandenburg area.  Using the flagging method, questing ticks were collected at four sites in 2010-2012 and 2000 D. reticulatus were analysed regarding infection with Rickettsia, Babesia, Borrelia and Anaplasmataceae by conventional or real-time PCR.  Dermacentor reticulatus showed a bimodal activity pattern: highest numbers of adult ticks were recorded between March and end of May (mean 50 ticks/h) and from mid-August until end of November (mean 102 ticks/h).  During summer, almost complete inactivity was observed (mean 0.4 ticks/h).  Sporadic samplings from December to February revealed tick activity also during winter (mean 47 ticks/h), which was characterised by large fluctuations.  Using negative binomial regression analysis, significant influences of the variables sampling site, season and temperature on the abundance of questing D. reticulatus were determined.  The parameters relative humidity and year were not of significant importance.  PCR analyses showed an average prevalence of 64% for Rickettsia sp.  Large differences in pathogen frequencies were observed between sampling sites (31.4-78.3%).  Regression analysis demonstrated a significant influence of the sampling site but not of season and year.  Examinations regarding other pathogen groups indicated prevalences of 0.25% (Borrelia sp.) and 0.05% (Anaplasmataceae) but absence of Babesia sp.  Sequencing of positive samples revealed infections with Rickettsia raoultii, Borrelia miyamotoi, Borrelia afzelii and Anaplasma phagocytophilum.  The study shows stable populations of D. reticulatus in Berlin/Brandenburg.  People should be aware of ticks throughout the year since Ixodes ricinus is co-endemic and active in spring, summer and autumn while adult D. reticulatus are active throughout the year and even in winter during periods of frost as long as it is warming up during the day.  Prevalence of R. raoultii in the present study is among the highest described for D. reticulatus.  Borrelia miyamotoi was detected for the first time in D. reticulatus, illustrating the importance of screening studies to evaluate the pathogen structure in D. reticulatus populations.
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