Analysis of Babesia bovis infection-induced gene expression changes in larvae from the cattle tick, Rhipicephalus (Boophilus) microplus

Particle and Fibre Toxicology

Kelly A Brayton⁵  Appolinaire Djikeng²  Vishvanath Nene²  Cedric Gondro³  Glen A Scoles⁶  Leo Saldivar⁴  Kylie G Bendele¹  Felix D Guerrero¹  Andrew M Heekin¹ 

[1] USDA-ARS, Knipling Bushland US Livestock Insect Research Laboratory, 2700 Fredericksburg Rd, Kerrville, TX, 78028, USA;International Livestock Research Institute (ILRI) and Biosciences eastern and central Africa (BecA) Hub, PO Box 30709, Nairobi, Kenya;The Institute for Genetics and Bioinformatics, University of New England, Armidale, NSW, 2351, Australia;Department of Mathematics, University of Texas at El Paso, El Paso, TX, 79968, USA;Program in Vector-Borne Diseases, Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, 99164, USA;USDA-ARS Animal Disease Research Unit, Pullman, WA, 99164, USA

关键词: Serine proteinase inhibitor;    Transcriptome;    Larva;    Babesia bovis;    Rhipicephalus (Boophilus) microplus;    Cattle tick;   

Others  :  1231726
DOI  :  10.1186/1756-3305-5-162

 received in 2012-05-24, accepted in 2012-07-26,  发布年份 2012

【 摘 要 】

Background

Cattle babesiosis is a tick-borne disease of cattle that has severe economic impact on cattle producers throughout the world’s tropical and subtropical countries. The most severe form of the disease is caused by the apicomplexan, Babesia bovis, and transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus, with the most prevalent species being Rhipicephalus (Boophilus) microplus. We studied the reaction of the R. microplus larval transcriptome in response to infection by B. bovis.

Methods

Total RNA was isolated for both uninfected and Babesia bovis-infected larval samples. Subtracted libraries were prepared by subtracting the B. bovis-infected material with the uninfected material, thus enriching for expressed genes in the B. bovis-infected sample. Expressed sequence tags from the subtracted library were generated, assembled, and sequenced. To complement the subtracted library method, differential transcript expression between samples was also measured using custom high-density microarrays. The microarray probes were fabricated using oligonucleotides derived from the Bmi Gene Index database (Version 2). Array results were verified for three target genes by real-time PCR.

Results

Ticks were allowed to feed on a B. bovis-infected splenectomized calf and on an uninfected control calf. RNA was purified in duplicate from whole larvae and subtracted cDNA libraries were synthesized from Babesia-infected larval RNA, subtracting with the corresponding uninfected larval RNA. One thousand ESTs were sequenced from the larval library and the transcripts were annotated. We used a R. microplus microarray designed from a R. microplus gene index, BmiGI Version 2, to look for changes in gene expression that were associated with infection of R. microplus larvae. We found 24 transcripts were expressed at a statistically significant higher level in ticks feeding upon a B. bovis-infected calf contrasted to ticks feeding on an uninfected calf. Six transcripts were expressed at a statistically significant lower level in ticks feeding upon a B. bovis-infected calf contrasted to ticks feeding on an uninfected calf.

Conclusion

Our experimental approaches yielded specific differential gene expression associated with the infection of R. microplus by B. bovis. Overall, an unexpectedly low number of transcripts were found to be differentially expressed in response to B. bovis infection. Although the BmiGI Version 2 gene index (http://compbio.dfci.harvard.edu/tgi/cgi-bin/tgi/gimain.pl?gudb=b_microplus webcite) was a useful database to help assign putative function to some transcripts, a majority of the differentially expressed transcripts did not have annotation that was useful for assignment of function and specialized bioinformatic approaches were necessary to increase the information from these transcriptome experiments.

【 授权许可】

   
2012 Heekin et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1 .

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