Olivieri Emanuela, Zanzani Sergio A, Latrofa Maria S, Lia Riccardo P, Dantas-Torres Filipe, Otranto Domenico, Manfredi Maria T
Department of Veterinary Medicine, University of Perugia, 06126, Perugia, Italy.
Department of Veterinary Medicine, University of Milan, 20133, Milan, Italy.
Parasit Vectors. 2016 Apr 18;9:213. doi: 10.1186/s13071-016-1502-9.
Two clustered clinical cases of canine babesiosis were diagnosed by veterinary practitioners in two areas of northeastern Italy close to natural parks. This study aimed to determine the seroprevalence of babesial infection in dogs, the etiological agents that cause canine babesiosis and the potential tick vector for the involved Babesia spp.
The study area was represented by two parks in northeastern Italy: Groane Regional Park (Site A) and the Ticino Valley Lombard Park (Site B). From March to May 2015 ticks were collected from the vegetation in three transects in each site. In the same period, blood samples were collected from 80 dogs randomly chosen from veterinary clinics and kennel located in the two areas. Morphological identification of the ticks was performed and six specimens were molecularly characterised by the amplification and sequencing of partial mitochondrial 12S rRNA, 16S rRNA and cox1 genes. For phylogenetic analyses, sequences herein obtained for all genes and those available from GenBank for other Dermacentor spp. were included. Dog serum samples were analysed with a commercial indirect fluorescent antibody test to detect the presence of IgG antibodies against Babesia canis. Ticks and blood samples were tested by PCR amplification using primers targeting 18S rRNA gene of Babesia spp.
Ticks collected (n = 34) were morphologically identified as adults of D. reticulatus. Twenty-eight ticks were found in all transects from Site A and the remaining six were collected in Site B. Blast analysis of mitochondrial sequences confirmed the morphological identification of processed tick specimens by revealing a highest nucleotide similarity (99-100%) with those of D. reticulatus available in the GenBank database. The phylogenetic trees were concordant in clustering D. reticulatus in a monophyletic clade. Seven dogs (8.8%) had antibodies against B. canis, most of which (n = 6) came from Site A. Analysis of nucleotide sequences obtained from one tick and from one dog identified B. canis displayed a 100% similarity to those available in GenBank.
This study morphologically and molecularly confirms the presence of D. reticulatus in Italy and links it, for the first time, with the occurrence of B. canis infection in dogs in this country.
意大利东北部靠近自然公园的两个地区的兽医从业者诊断出两例聚集性犬巴贝斯虫病临床病例。本研究旨在确定犬类中巴贝斯虫感染的血清阳性率、引起犬巴贝斯虫病的病原体以及所涉及的巴贝斯虫属的潜在蜱传播媒介。
研究区域以意大利东北部的两个公园为代表：格罗阿内地区公园（地点A）和提契诺山谷伦巴第公园（地点B）。2015年3月至5月，在每个地点的三个样带中从植被上采集蜱虫。同一时期，从位于这两个地区的兽医诊所和犬舍中随机选取80只狗采集血样。对蜱虫进行形态学鉴定，并通过对部分线粒体12S rRNA、16S rRNA和cox1基因进行扩增和测序，对六个标本进行分子特征分析。为进行系统发育分析，纳入了本文获得的所有基因序列以及从GenBank获得的其他硬蜱属物种的序列。用商业间接荧光抗体试验分析犬血清样本，以检测抗犬巴贝斯虫IgG抗体的存在。使用针对巴贝斯虫属18S rRNA基因的引物通过PCR扩增对蜱虫和血样进行检测。
采集的蜱虫（n = 34）经形态学鉴定为网纹硬蜱成虫。在地点A的所有样带中发现了28只蜱虫，其余6只在地点B采集。线粒体序列的Blast分析通过揭示与GenBank数据库中可用的网纹硬蜱序列具有最高核苷酸相似性（99 - 100%），证实了处理后的蜱虫标本的形态学鉴定。系统发育树在将网纹硬蜱聚类到一个单系类群中是一致的。7只狗（8.8%）具有抗犬巴贝斯虫抗体，其中大多数（n = 6）来自地点A。对从一只蜱虫和一只狗获得的核苷酸序列进行分析，鉴定出犬巴贝斯虫与GenBank中可用的序列具有100%的相似性。
本研究从形态学和分子学上证实了意大利存在网纹硬蜱，并首次将其与该国犬类中犬巴贝斯虫感染的发生联系起来。
