Approaches for Reverse Line Blot-Based Detection of Microbial Pathogens in Ixodes ricinus Ticks Collected in Austria and Impact of the Chosen Method

作者：Schoetta Anna Margarita; Wijnveld Michiel; Stockinger Hannes; Stanek Gerold

来源：Applied and Environmental Microbiology, 2017, 83(13): UNSP e00489-17.

DOI：10.1128/AEM.00489-17

<jats:title>ABSTRACT</jats:title>
<jats:p>
Ticks transmit a large number of pathogens capable of causing human disease. In this study, the PCR-reverse line blot (RLB) method was used to screen for pathogens in a total of 554
<jats:named-content content-type="genus-species">Ixodes ricinus</jats:named-content>
ticks collected from all provinces of Austria. These pathogens belong to the genera
<jats:named-content content-type="genus-species">Borrelia</jats:named-content>
,
<jats:named-content content-type="genus-species">Rickettsiae</jats:named-content>
,
<jats:named-content content-type="genus-species">Anaplasma</jats:named-content>
/
<jats:named-content content-type="genus-species">Ehrlichia</jats:named-content>
(including “
<jats:named-content content-type="genus-species">Candidatus</jats:named-content>
Neoehrlichia”),
<jats:named-content content-type="genus-species">Babesia</jats:named-content>
, and
<jats:named-content content-type="genus-species">Coxiella</jats:named-content>
. The pathogens with the highest detected prevalence were spirochetes of the
<jats:named-content content-type="genus-species">Borrelia burgdorferi</jats:named-content>
<jats:named-content content-type="genus-species">sensu lato</jats:named-content>
complex, in 142 ticks (25.6%).
<jats:named-content content-type="genus-species">Borrelia afzelii</jats:named-content>
(80/142) was the most frequently detected species, followed by
<jats:named-content content-type="genus-species">Borrelia burgdorferi</jats:named-content>
<jats:named-content content-type="genus-species">sensu stricto</jats:named-content>
(38/142) and
<jats:named-content content-type="genus-species">Borrelia valaisiana</jats:named-content>
(36/142).
<jats:named-content content-type="genus-species">Borrelia garinii/Borrelia bavariensis</jats:named-content>
,
<jats:named-content content-type="genus-species">Borrelia lusitaniae</jats:named-content>
, and
<jats:named-content content-type="genus-species">Borrelia spielmanii</jats:named-content>
were found in 28 ticks, 5 ticks, and 1 tick, respectively.
<jats:named-content content-type="genus-species">Rickettsia</jats:named-content>
spp. were detected in 93 ticks (16.8%):
<jats:named-content content-type="genus-species">R. helvetica</jats:named-content>
(39/93),
<jats:named-content content-type="genus-species">R. raoultii</jats:named-content>
(38/93),
<jats:named-content content-type="genus-species">R. monacensis</jats:named-content>
(2/93), and
<jats:named-content content-type="genus-species">R. slovaca</jats:named-content>
(1/93). Thirteen
<jats:named-content content-type="genus-species">Rickettsia</jats:named-content>
samples remain uncharacterized. “
<jats:named-content content-type="genus-species">Candidatus</jats:named-content>
Neoehrlichia mikurensis,”
<jats:named-content content-type="genus-species">Babesia</jats:named-content>
spp. (
<jats:named-content content-type="genus-species">B. venatorum</jats:named-content>
,
<jats:named-content content-type="genus-species">B. divergens</jats:named-content>
,
<jats:named-content content-type="genus-species">B. microti</jats:named-content>
), and
<jats:named-content content-type="genus-species">Anaplasma phagocytophilum</jats:named-content>
were found in 4.5%, 2.7%, and 0.7%, respectively.
<jats:named-content content-type="genus-species">Coxiella burnetii</jats:named-content>
was not detected. Multiple microorganisms were detected in 40 ticks (7.2%), and the cooccurrence of
<jats:named-content content-type="genus-species">Babesia</jats:named-content>
spp. and “
<jats:named-content content-type="genus-species">Candidatus</jats:named-content>
Neoehrlichia mikurensis” showed a significant positive correlation. We also compared different PCR-RLBs for detection of
<jats:named-content content-type="genus-species">Borrelia burgdorferi</jats:named-content>
<jats:named-content content-type="genus-species">sensu lato</jats:named-content>
and
<jats:named-content content-type="genus-species">Rickettsia</jats:named-content>
spp. and showed that different detection approaches provide highly diverse results, indicating that analysis of environmental samples remains challenging.
</jats:p>
<jats:p>
<jats:bold>IMPORTANCE</jats:bold>
This study determined the wide spectrum of tick-borne bacterial and protozoal pathogens that can be encountered in Austria. Surveillance of (putative) pathogenic microorganisms occurring in the environment is of medical importance, especially when those agents can be transmitted by ticks and cause disease. The observation of significant coinfections of certain microorganisms in field-collected ticks is an initial step to an improved understanding of microbial interactions in ticks. In addition, we show that variations in molecular detection methods, such as in primer pairs and target genes, can considerably influence the final results. For instance, detection of certain genospecies of borreliae may be better or worse by one method or the other, a fact of great importance for future screening studies.
</jats:p>

• 出版日期2017-7