Grillini Marika, Beraldo Paola, Frangipane di Regalbono Antonio, Dotto Giorgia, Tessarin Cinzia, Franzo Giovanni, Marchiori Erica, Modrý David, Simonato Giulia
Department of Animal Medicine, Production and Health, University of Padua, Padua, Italy.
Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, Udine, Italy.
Front Vet Sci. 2023 Jun 12;10:1113681. doi: 10.3389/fvets.2023.1113681. eCollection 2023.
Tick-transmitted apicomplexans of the genera and affect a wide range of felids worldwide, but little is known about them. Recently, several studies addressed the species circulating in Europe, their distribution, and their hosts. Molecular assays are the method of choice for their detection. Unfortunately, conventional PCRs already described are time- and cost-consuming and specific for either or detection. This study was developed to evaluate (i) the occurrence of and in felids using a fast and cost-saving real-time PCR capable of detecting both protozoa simultaneously, (ii) the distribution of and species in north-eastern Italy, and (iii) the involvement of other susceptible felid hosts in the same area. An SYBR Green-based real-time PCR with primers targeting the 18S-rRNA was validated and applied to 237 felid samples, i.e., whole blood from 206 domestic cats and 12 captive exotic felids, and tissues from 19 wildcats. Positive results were obtained by melting temperature curve analysis due to the specific melting peak (i.e., 81°C spp.; 78-78.5°C spp.). Positive samples were subjected to conventional PCR, followed by sequencing for species identification. Phylogenetic analyses were performed to assess relatedness among European isolates. Data on domestic cats (age class, sex, origin, management, and lifestyle) were recorded, and statistical analyses were performed to identify potential risk factors. A total of 31 (15%) domestic cats were positive for spp. (i.e., 12 for 19 for ), while six (2.9%) for . The prevalence of was significantly ( < 0.05) higher in domestic cats, while was higher in strays and animals from the Eastern region (i.e., Friuli-Venezia Giulia). was detected only in stray cats from Friuli-Venezia Giulia (province of Trieste). Among captive felids, one tiger was infected with and another with ; eight out of 19 (42%) wildcats were positive for spp. (i.e., six with , two with ) and four out of 19 (21%) for . Outdoor lifestyle and origin (i.e., Friuli-Venezia Giulia region) were the most relevant risk factors for and infections. Conversely, was most frequently isolated from domestic cats, suggesting different modes of transmission.
蜱传播的巴贝斯属和泰勒虫属顶复门原虫在全球范围内影响着广泛的猫科动物，但人们对它们了解甚少。最近，几项研究涉及了在欧洲传播的这些物种、它们的分布以及宿主。分子检测是检测它们的首选方法。不幸的是，已描述的传统聚合酶链反应（PCR）既耗时又费钱，且分别针对巴贝斯虫或泰勒虫的检测具有特异性。本研究旨在评估：（i）使用一种能够同时检测这两种原生动物的快速且节省成本的实时PCR，检测猫科动物中巴贝斯虫和泰勒虫的存在情况；（ii）巴贝斯虫和泰勒虫物种在意大利东北部的分布；（iii）同一地区其他易感猫科动物宿主的感染情况。一种基于SYBR Green的实时PCR，其引物靶向18S核糖体RNA（rRNA），经过验证后应用于237份猫科动物样本，即206只家猫和12只圈养外来猫科动物的全血，以及19只野猫的组织样本。通过熔解温度曲线分析获得了阳性结果，这是由于特定的熔解峰（即，巴贝斯虫属为81°C；泰勒虫属为78 - 78.5°C）。对阳性样本进行传统PCR，随后进行测序以进行物种鉴定。进行系统发育分析以评估欧洲分离株之间的亲缘关系。记录了家猫的数据（年龄组、性别、来源、管理方式和生活方式），并进行统计分析以确定潜在的风险因素。共有31只（15%）家猫的巴贝斯虫属检测呈阳性（即，12只为巴贝斯虫，19只为分歧巴贝斯虫），而泰勒虫属为6只（2.9%）。家猫中巴贝斯虫属的患病率显著更高（P < 0.05），而泰勒虫属在流浪猫和来自东部地区（即弗留利 - 威尼斯朱利亚）的动物中患病率更高。泰勒虫仅在弗留利 - 威尼斯朱利亚（的里雅斯特省）的流浪猫中检测到。在圈养猫科动物中，一只老虎感染了巴贝斯虫，另一只感染了泰勒虫；19只野猫中有8只（42%）的巴贝斯虫属检测呈阳性（即，6只为巴贝斯虫，2只为分歧巴贝斯虫），19只中有4只（21%）的泰勒虫属检测呈阳性。户外生活方式和来源（即弗留利 - 威尼斯朱利亚地区）是巴贝斯虫和泰勒虫感染最相关的风险因素。相反，泰勒虫最常从家猫中分离出来，这表明传播方式不同。
