Antunes Sandra, Merino Octávio, Lérias Joana, Domingues Nuno, Mosqueda Juan, de la Fuente José, Domingos Ana
Institute of Hygiene and Tropical Medicine, New University of Lisbon, Rua da Junqueira 100, 1349-008 Lisboa, Portugal.
SaBio, Research Institute on Cinegetic Resources, IREC-CSIC-UCLM-JCCM, Ronda de Toledo s/n, 13005 Ciudad Real, Spain.
Ticks Tick Borne Dis. 2015 Feb;6(1):47-55. doi: 10.1016/j.ttbdis.2014.09.003. Epub 2014 Sep 26.
Ticks are obligate haematophagous ectoparasites considered the principal vectors of disease among animals. Rhipicephalus microplus and R. annulatus ticks are the most important vectors for Babesia bigemina and B. bovis, two of the most important intraerythrocytic protozoan parasites species in cattle, responsible for babesiosis which together with anaplasmosis account for substantial economic losses in the livestock industry worldwide. Anti-tick vaccines are a proved alternative to traditional tick and tick borne diseases control methods but are still limited primarily due to the lack of effective antigens. Subsequently to the identification of antigens the validation is a laborious work often expensive. Tick artificial feeding, is a low cost alternative to test antigens allowing achieving critical data. Herein, R. microplus females were successfully artificially fed using capillary tubes. Calreticulin (CRT) protein, which in a previous study has been identified as being involved in B. bigemina infection in R. annulatus ticks, was expressed as recombinant protein (rCRT) in an E. coli expression system and antibodies raised against rCRT. Anti-rCRT serum was supplemented to a blood meal, offered to partially engorged R. microplus females and their effect in feeding process as well as infection by B. bigemina was analyzed. No significant reductions in tick and egg weight were observed when ticks fed with anti-rCRT serum. Furthermore, B. bigemina infection levels did not show a statistically significant decrease when ticks fed with anti-rCRT antibodies. Results suggest that CRT is not a suitable candidate for cattle vaccination trials.
蜱是专性吸血的体外寄生虫,被认为是动物疾病的主要传播媒介。微小牛蜱和环形牛蜱是双芽巴贝斯虫和牛巴贝斯虫最重要的传播媒介,这两种是牛体内最重要的红细胞内原生动物寄生虫,可导致巴贝斯虫病,与无浆体病一起给全球畜牧业造成重大经济损失。抗蜱疫苗是传统蜱及蜱传疾病控制方法的一种经证实的替代方法,但主要仍受到缺乏有效抗原的限制。在鉴定出抗原之后,验证工作既费力又昂贵。蜱的人工饲养是一种低成本的抗原测试替代方法,能够获取关键数据。在此,使用毛细管成功地对微小牛蜱雌蜱进行了人工饲养。钙网蛋白(CRT)蛋白在先前的一项研究中已被确定与环形牛蜱感染双芽巴贝斯虫有关,在大肠杆菌表达系统中作为重组蛋白(rCRT)表达,并产生了针对rCRT的抗体。将抗rCRT血清添加到血餐中,提供给部分饱血的微小牛蜱雌蜱,并分析其在进食过程中的作用以及对双芽巴贝斯虫感染的影响。当蜱用抗rCRT血清喂养时,未观察到蜱和卵的重量有显著减轻。此外,当蜱用抗rCRT抗体喂养时,双芽巴贝斯虫的感染水平没有显示出统计学上的显著下降。结果表明,CRT不是牛疫苗接种试验的合适候选抗原。