Šlapeta Štěpán, Šlapeta Jan
School of Life and Environmental Sciences, Faculty of Veterinary Science, McMaster Building B14, The University of Sydney, New South Wales 2006, Australia.
School of Life and Environmental Sciences, Faculty of Veterinary Science, McMaster Building B14, The University of Sydney, New South Wales 2006, Australia.
Vet Parasitol Reg Stud Reports. 2016 Jun;3-4:36-40. doi: 10.1016/j.vprsr.2016.06.005. Epub 2016 Jul 2.
The cat flea (Ctenocephalides felis) is the most common ectoparasite of dogs and cats. Close association of humans with cats and dogs enables flea-borne disease transmission either directly, via flea bites, or indirectly, via pathogens excreted in flea faeces. The aim of this study was to evaluate molecular identity of C. felis from cats in Georgia, USA based on a molecular barcode marker gene (cox1) and the frequency at which the fleas were carriers of the vector-borne bacteria, Bartonella and Rickettsia. The multiplexed Bartonella and Rickettsia real-time PCR assay (targeting ssrA and gltA, respectively) adopted in this study, together with sequencing of the ssrA enabled rapid identification of Bartonella spp. in cat fleas. Eighteen out of 20 fleas examined were positive for Bartonella spp. and all fleas were positive for Rickettsia spp. DNA sequencing of the ssrA confirmed presence B. clarridgeiae and B. henselae. Amplification and DNA sequencing of gltA and ompA confirmed presence of Rickettsia sp. RF2125 (Rickettsia felis-like organism). All fleas from 21 cats in Georgia, USA were morphologically identical with C. felis. Sequencing of the flea cox1 revealed identity with C. felis from Seychelles, Africa recognised as a subspecies C. felis strongylus, the African cat flea. Analysis of the cox1 sequences of fleas improves understanding of global distribution of cat flea cox1 clades (C. felis) when compared with sequences from Ctenocephalides spp. from Asia, Africa, Europe, Asia and Australia. Coupling flea barcoding approach with the multiplexed real-time PCR assay followed by Bartonella sequencing represents a rational approach for screening and elucidation of fleas' capacity to vector infectious agents.
猫蚤（猫栉首蚤）是犬猫最常见的体外寄生虫。人类与猫和狗的密切接触使得跳蚤传播的疾病能够直接通过跳蚤叮咬传播，或者间接通过跳蚤粪便中排出的病原体传播。本研究的目的是基于分子条形码标记基因（细胞色素氧化酶亚基I，cox1）评估美国佐治亚州猫身上猫栉首蚤的分子特征，以及这些跳蚤作为媒介传播细菌（巴尔通体和立克次体）携带者的频率。本研究采用的多重巴尔通体和立克次体实时荧光定量PCR检测方法（分别针对单链核糖体RNA基因，ssrA和柠檬酸合酶基因，gltA），以及对ssrA进行测序，能够快速鉴定猫蚤中的巴尔通体菌种。在检测的20只跳蚤中，有18只巴尔通体菌种呈阳性，所有跳蚤立克次体菌种均呈阳性。对ssrA进行DNA测序证实存在克拉氏巴尔通体和汉赛巴尔通体。对gltA和外膜蛋白A（ompA）进行扩增和DNA测序证实存在立克次体sp. RF2125（类猫立克次体）。美国佐治亚州21只猫身上的所有跳蚤在形态上均与猫栉首蚤相同。对跳蚤cox1进行测序发现，其与来自非洲塞舌尔的猫栉首蚤相同，该猫栉首蚤被认定为猫栉首蚤亚种——非洲猫蚤（C. felis strongylus）。与来自亚洲、非洲、欧洲和澳大利亚的栉首蚤属（Ctenocephalides spp.）序列相比，对跳蚤cox1序列的分析有助于更好地理解猫蚤cox1进化枝（猫栉首蚤）的全球分布情况。将跳蚤条形码方法与多重实时荧光定量PCR检测方法相结合，随后对巴尔通体进行测序，是一种合理的方法，可用于筛选和阐明跳蚤传播感染因子的能力。
