Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Bock R, Jackcon L, de Vos A, Jorgensen W (2004) Babesiosis of cattle. Parasitology 129:S247–S269
de la Fuente J, Moreno-Cid JA, Canales M et al (2011) Targeting arthropod subolesin/akirin for the development of a universal vaccine for control of vector infestations and pathogen transmission. Vet Parasitol 181:17–22
Macaluso KR, Mulenga A, Simser JA et al (2003) Differential expression of genes in uninfected and rickettsia-infected Dermacentor variabilis ticks as assessed by differential-display PCR. Infect Immun 71:6165–6170
Mulenga A, Macaluso KR, Simser JA et al (2003) Dynamics of Rickettsia-tick interactions: identification and characterization of differentially expressed mRNAs in uninfected and infected Dermacentor variabilis. Insect Mol Biol 12:185–193
Nene V, Lee D, Kang’a S, Skilton R et al (2004) Genes transcribed in the salivary glands of female Rhipicephalus appendiculatus ticks infected with Theileria parva. Insect Biochem Mol Biol 34:1117–1128
Rudenko N, Golovchenko M, Edwards MJ et al (2005) Differential expression of Ixodes ricinus tick genes induced by blood feeding or Borrelia burgdorferi infection. J Med Entomol 42:36–41
de la Fuente J, Blouin EF, Manzano-Roman NR et al (2007) Functional genomic studies of tick cells in response to infection with the cattle pathogen, Anaplasma marginale. Genomics 90:712–722
de la Fuente J, Kocan KM, Almazan C et al (2007) RNA interference for the study and genetic manipulation of ticks. Trends Parasitol 23:427–433
Villar M, Torina A, Nunez Y et al (2010) Application of highly sensitive saturation labeling to the analysis of differential protein expression in infected ticks from limited samples. Proteome Sci 8:43
Zivkovic Z, Esteves E, Almazan C et al (2010) Differential expression of genes in salivary glands of male Rhipicephalus (Boophilus) microplus in response to infection with Anaplasma marginale. BMC Genomics 11:186
Mercado-Curiel RF, Palmer GH, Guerrero FD et al (2011) Temporal characterisation of the organ-specific Rhipicephalus microplus transcriptional response to Anaplasma marginale infection. Int J Parasitol 41:851–860
Antunes S, Golovchenko M, Rudenko N et al (2012) Gene silencing of the tick antigens selected after infection with Babesia bigemina, in the host tick Rhipicephalus annulatus by RNA interference. Int J Parasitol 42:187–195
Zivkovic Z, Torina A, Mitra R et al (2010) Subolesin expression in response to pathogen infection in ticks. BMC Immunol 11:7
de la Fuente J, Maritz-Olivier C, Naranjo V et al (2008) Evidence of the role of tick subolesin in gene expression. BMC Genomics 9:372
Kocan KM, Zivkovic Z, Blouin EF et al (2009) Silencing of genes involved in Anaplasma marginale–tick interactions affects the pathogen developmental cycle in Dermacentor variabilis. BMC Dev Biol 9:42
Guo YJ, Ribeiro JMC, Anderson JM et al (2009) dCAS: a desktop application for cDNA sequence annotation. Bioinformatics 25:1195–1196
Fire A, Xu S, Montgomery MK, Kostas SA et al (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811
Montgomery MK, Xu SQ, Fire A (1998) RNA as a target of double-stranded RNA-mediated genetic interference in Caenorhabditis elegans. Proc Natl Acad Sci U S A 95:15502–15507
Mello CC, Conte D (2004) Revealing the world of RNA interference. Nature 431:338–342
de la Fuente J, Kocan KM (2006) Strategies for development of vaccines for control of ixodid tick species. Parasite Immunol 28:275–283
Hoa NT, Keene KM, Olson KE et al (2003) Characterization of RNA interference in an Anopheles gambiae cell line. Insect Biochem Mol Biol 33:949–957
Pal U, Li X, Wang T et al (2004) TROSPA, an Ixodes scapularis receptor for Borrelia burgdorferi. Cell 119:457–468
Ramamoorthi N, Narasimhan S, Pal U et al (2005) The Lyme disease agent exploits a tick protein to infect the mammalian host. Nature 436:573–577
Sukumaran B, Narasimhan S, Anderson JF et al (2006) An Ixodes scapularis protein required for survival of Anaplasma phagocytophilum in tick salivary glands. J Exp Med 203:1507–1517
Merino O, Almazan C, Canales M et al (2011) Control of Rhipicephalus (Boophilus) microplus infestations by the combination of subolesin vaccination and tick autocidal control after subolesin gene knockdown in ticks fed on cattle. Vaccine 29:2248–2254
Shkap V, Leibovitz B, Krigel Y et al (2005) Vaccination of older Bos taurus bulls against bovine babesiosis. Vet Parasitol 129:235–242
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods 25:402–408
Schefe JH, Lehmann KE, Buschmann IR et al (2006) Quantitative real-time RT-PCR data analysis: current concepts and the novel “gene expression's C-T difference” formula. J Mol Med 84:901–910
de la Fuente J, Almazan C, Blouin EF et al (2006) Reduction of tick infections with Anaplasma marginale and A. phagocytophilum by targeting the tick protective antigen subolesin. Parasitol Res 100:85–91
Willadsen P, Kemp DH (1988) Vaccination with concealed antigens for tick control. Parasitol Today 4:196–198
Suarez CE, Palmer GH, Florin-Christensen M et al (2003) Organization, transcription, and expression of rhoptry associated protein genes in the Babesia bigemina rap-1 locus. Mol Biochem Parasitol 127:101–112
Petrigh R, Ruybal P, Thompson C et al (2008) Improved molecular tools for detection of Babesia bigemina. Ann N Y Acad Sci 1149:155–157
Bernard P, Gabant P, Bahassi EM, Couturier M (1994) Positive-selection vectors using the F plasmid ccdB killer gene. Gene 148:71–74
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Domingos, A., Antunes, S., Villar, M., de la Fuente, J. (2015). Functional Genomics of Tick Vectors Challenged with the Cattle Parasite Babesia bigemina . In: Cunha, M., Inácio, J. (eds) Veterinary Infection Biology: Molecular Diagnostics and High-Throughput Strategies. Methods in Molecular Biology, vol 1247. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2004-4_32
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2004-4_32
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2003-7
Online ISBN: 978-1-4939-2004-4
eBook Packages: Springer Protocols