Domingos Ana, Antunes Sandra, Villar Margarita, de la Fuente José
Global Health and Tropical Medicine, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, Portugal,
Methods Mol Biol. 2015;1247:475-89. doi: 10.1007/978-1-4939-2004-4_32.
Ticks are obligate hematophagous ectoparasites considered as vectors of animal diseases, having a huge economic impact in cattle industry. Babesia spp. are tick-borne pathogens that cause a disease called babesiosis in a wide range of animals and in humans. Control of tick infestations is mainly based on the use of acaricides, which have limited efficacy reducing tick infestations, mostly due to wrong usage, and is often accompanied by the selection of acaricide-resistant ticks, environmental contamination, and contamination of milk and meat products. Vaccines affecting both vector and pathogens constitute new control strategies for tick and tick-borne diseases and are, therefore, a good alternative to chemical control. In this chapter we describe the identification of Rhipicephalus (Boophilus) annulatus genes differentially expressed in response to infection with B. bigemina by using suppression-subtractive hybridization (SSH), which allows the identification of differentially expressed genes. The results of the SSH studies are validated by real-time reverse transcription (RT)-PCR. Functional analyses are conducted by RNAi on selected R. annulatus genes to determine their putative role in B. bigemina-tick interactions. Gathered data may be useful for the future development of improved vaccines and vaccination strategies to control babesiosis.
蜱是专性吸血的体外寄生虫，被视为动物疾病的传播媒介，对养牛业造成巨大经济影响。巴贝斯虫属是蜱传播的病原体，可在多种动物和人类中引起一种名为巴贝斯虫病的疾病。控制蜱虫感染主要基于使用杀螨剂，但杀螨剂减少蜱虫感染的效果有限，主要原因是使用不当，而且往往会导致产生抗杀螨剂的蜱虫、环境污染以及牛奶和肉类产品受到污染。影响传播媒介和病原体的疫苗构成了蜱虫及蜱传疾病的新控制策略，因此是化学控制的良好替代方法。在本章中，我们描述了通过抑制性消减杂交（SSH）鉴定微小牛蜱中因感染双芽巴贝斯虫而差异表达的基因，该方法可用于鉴定差异表达基因。SSH研究结果通过实时逆转录（RT）-PCR进行验证。通过RNA干扰对选定的微小牛蜱基因进行功能分析，以确定它们在双芽巴贝斯虫与蜱虫相互作用中的假定作用。收集的数据可能有助于未来开发改进的疫苗和疫苗接种策略以控制巴贝斯虫病。
