Detection and differentiation of Borrelia burgdorferi sensu lato in ticks collected from sheep and cattle in China

<p>Abstract</p> <p>Background</p> <p>Lyme disease caused by <it>Borrelia burgdorferi </it>sensu lato complex is an important endemic zoonosis whose distribution is closely related to the main ixodid tick vectors. In China, isolated cases of Lyme disease infection of humans have been reported in 29 provinces. Ticks, especially ixodid ticks are abundant and a wide arrange of <it>Borrelia </it>natural reservoirs are present. In this study, we developed a reverse line blot (RLB) to identify <it>Borrelia </it>spp. in ticks collected from sheep and cattle in 7 Provinces covering the main extensive livestock regions in China.</p> <p>Results</p> <p>Four species-specific RLB oligonucleotide probes were deduced from the spacer region between the 5S-23S rRNA gene, along with an oligonucleotide probe which was common to all. The species specific probes were shown to discriminate between four genomic groups of <it>B. burgdorferi </it>sensu lato i.e. <it>B. burgdorferi </it>sensu stricto, <it>B. garinii, B. afzelii</it>, and <it>B. valaisiana</it>, and to bind only to their respective target sequences, with no cross reaction to non target DNA. Furthermore, the RLB could detect between 0.1 pg and 1 pg of <it>Borrelia </it>DNA.</p> <p>A total of 723 tick samples (<it>Haemaphysalis, Boophilus, Rhipicephalus </it>and <it>Dermacentor</it>) from sheep and cattle were examined with RLB, and a subset of 667 corresponding samples were examined with PCR as a comparison. The overall infection rate detected with RLB was higher than that of the PCR test.</p> <p>The infection rate of <it>B. burgdoreri </it>sensu stricto was 40% in south areas; while the <it>B. garinii infection rate </it>was 40% in north areas. The highest detection rates of <it>B. afzelii </it>and <it>B. valaisiana </it>were 28% and 22%, respectively. Mixed infections were also found in 7% of the ticks analyzed, mainly in the North. The proportion of <it>B. garinii </it>genotype in ticks was overall highest at 34% in the whole investigation area.</p> <p>Conclusion</p> <p>In this study, the RLB assay was used to detect <it>B. burgdorferi </it>sensu lato in ticks collected from sheep and cattle in China. The results showed that <it>B. burdorferi senso stricto </it>and <it>B. afzelii </it>were mainly distributed in the South; while <it>B. garinii </it>and <it>B. valaisiana </it>were dominant in the North. <it>Borrelia </it>spirochaetes were detected in <it>Rhipicephalus </it>spp for the first time. It is suggested that the <it>Rhipicephalus </it>spps might play a role in transmitting <it>Borrelia </it>spirochaetes.</p

Theileria annulata Cyclophilin1 (TaCyp1) Interacts With Host Cell MED21

Host cells infected by Theileria annulata schizonts show the character of permanent proliferation in vitro, also named transformation. To explore the molecular mechanism a T. annulata Cyp1 (TaCyp1) protein potentially involved in regulating cell transformation was used as bait to screen for its interacting proteins by yeast-two-hybrid assay. Additional GST-pull down experiments confirmed that only MED21 specifically interacted with TaCyp1. Moreover, the distribution of TaCyp1 around T. annulata schizonts facilitated interaction with host cell MED21. As a component of mediator complex, MED21 is normally involved in regulating the transcription of nearly all RNA polymerase II-dependent genes. Therefore, to explore its influence on NF-κB signaling MED21 RNA interference and parasite killing with BW720c treatment were performed. Knock down of MED21 resulted in a significant decrease in NF-κB1/2 mRNA expressions, but no significant change in P105, P52 levels, nor detectable alteration in levels of phosphorylated IκBα/β. By contrast, BW720c treatment induced an obvious decrease in the phosphorylation status of P52 and IκBα/β, but no obvious change in that of P105. This suggests that BW720c-induced parasite death had a significant negative influence on NF-κB signaling, whereas knock down of MED21 had no obvious effect on NF-κB signaling. Characterization of TaCyp1 provides information on the function of parasite cyclophilins and leads to a better understanding of the interactions between T. annulata and its host leukocytes

Molecular Evidence of Bartonella melophagi in Ticks in Border Areas of Xinjiang, China

Bartonella are gram-negative intracellular bacteria; certain species of Bartonella can cause diseases in mammals and humans. Ticks play a major role in the transmission of Bartonella. Xinjiang is the largest province in China according to land area and has one-third of the tick species in China; the infection rate of Bartonella in ticks in the Xinjiang border areas has not been studied in detail. Therefore, this study investigated tick infections by Bartonella in Xinjiang border areas, and the purpose of the study was to fill in gaps in information regarding the genetic diversity of tick infections by Bartonella in Xinjiang. We tested 1,549 tick samples from domestic animals (sheep and cattle) for Bartonella using ribC-PCR. Positive samples from the ribC-PCR assay for Bartonella spp. were further subjected to PCR assays targeting the ITS, rpoB and gltA genes followed by phylogenetic analyses. Bartonella DNA was detected in 2.19% (34/1,549) of tick samples, and the ITS, rpoB and gltA genes of ribC gene-positive samples were amplified to identify nine samples of Bartonella melophagi. In this study, molecular analysis was used to assess the presence and genetic diversity of B. melophagi in ticks collected from sheep and cattle from Xinjiang, China. This study provides new information on the presence and identity of B. melophagi in ticks from sheep and cattle