Differential Expression of Three Members of the Multidomain Adhesion CCp Family in Babesia bigemina, Babesia bovis and Theileria equi

Bastos, Reginaldo G.; Suárez, Carlos E.; Laughery, Jacob M.; Johnson, Wendell C.; Ueti, Massaro W.; Knowles, Donald P.

doi:10.1371/journal.pone.0067765

Cited by 39 publications

(60 citation statements)

References 38 publications

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“…Gene expression was normalized to the total amount of RNA used to generate the cDNA, as xpreviously described [ 13 ]. The RmAQP2 transcription level was then calculated as a relative expression using the formula: Relative expression (sample) = 2 [ Cq (control) − Cq (sample)] , where the control is either the highest or lowest C q value for the gene of interest, as previously described [ 13 , 25 ]. Melt curve analyses showed the absence of primer dimers and nonspecific amplification.…”

Section: Methodsmentioning

confidence: 99%

Targeted silencing of the Aquaporin 2 gene of Rhipicephalus (Boophilus) microplus reduces tick fitness

et al. 2015

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BackgroundTicks are blood-feeding arthropods that can affect human and animal health both directly by blood-feeding and indirectly by transmitting pathogens. The cattle tick Rhipicephalus (Boophilus) microplus is one of the most economically important ectoparasites of bovines worldwide and it is responsible for the transmission of the protozoan Babesia bovis, the etiological agent of bovine babesiosis. Aquaporins (AQPs) are water channel proteins implicated in physiological mechanisms of osmoregulation. Members of the AQP family are critical for blood-feeding arthropods considering the extreme osmoregulatory changes that occur during their feeding. We investigated the pattern of expression of a newly identified AQP2 gene of R. microplus (RmAQP2) in different tick tissues and stages. We also examined in vivo the biological implications of silencing expression of RmAQP2 silencing during tick feeding on either uninfected or B. bovis-infected cattle.MethodsIn silico gene analyses were performed by multiple alignments of amino acid sequences and topology prediction. Levels of RmAQP2 transcripts in different tick tissues and stages were analyzed by reverse transcriptase quantitative PCR. Patterns of expression of RmAQP2 protein were investigated by immunoblots. Gene silencing was performed by RNA interference and in vivo functional analyses carried out by feeding ticks on either uninfected or B. bovis-infected cattle.ResultsRmAQP2 transcripts were found in unfed larvae, engorged nymphs, and salivary glands and guts of partially engorged females; however, of all tick tissues and stages examined, RmAQP2 protein was found only in salivary glands of partially engorged females. RmAQP2 silencing significantly reduced tick fitness and completely abrogated protein expression. The effect of RmAQP2 silencing on fitness was more pronounced in females fed on a B. bovis-infected calf than in ticks fed on an uninfected calf and none of their larval progeny survived.ConclusionsCollectively, considering the gene expression and tick fitness data, we conclude that RmAQP2 is critical for tick blood feeding and may be a suitable candidate target for the development of novel strategies to control R. microplus and tick-borne parasites.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-1226-2) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Targeted silencing of the Aquaporin 2 gene of Rhipicephalus (Boophilus) microplus reduces tick fitness

et al. 2015

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“…CCp proteins are, in general, characterized by the presence of at least one Limulus coagulation factor C (LCCL) domain [ 15 , 16 ] and are often involved in cell adhesion [ 16 ]. Gene orthologs from the highly-conserved CCp family have been identified in numerous apicomplexan parasites [ 16 – 18 ], including Babesia and Theileria species [ 14 , 19 ]. Transcription of ccp genes was found to be restricted to gametocytes in vertebrate blood, while translation occurs in the arthropod vector to mediate gamete fertilization [ 14 , 16 , 20 – 24 ].…”

Section: Introductionmentioning

confidence: 99%

“…Based on post-genomic bioinformatic analyses of Babesia and Plasmodium genomes, three bdccp genes ( bdccp1 , bdccp2 and bdccp3 ) were thoroughly characterized and described as markers of B. divergens sexual stages [ 14 ]. The transcripts of bdccp1 , bdccp2 and bdccp3 genes were also detected in gametocytes appearing in cultures of B. divergens , B. bigemina , Babesia bovis and Theileria equi [ 14 , 19 ]. Moreover, antibody targeted to BdCCp2 protein enabled visualization of B. divergens sexual stages exclusively in the midgut of Ixodes ricinus [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

Stimulation and quantification of Babesia divergens gametocytogenesis

et al. 2016

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BackgroundBabesia divergens is the most common blood parasite in Europe causing babesiosis, a tick-borne malaria-like disease. Despite an increasing focus on B. divergens, especially regarding veterinary and human medicine, the sexual development of Babesia is poorly understood. Development of Babesia sexual stages in the host blood (gametocytes) plays a decisive role in parasite acquisition by the tick vector. However, the exact mechanism of gametocytogenesis is still unexplained.MethodsBabesia divergens gametocytes are characterized by expression of bdccp1, bdccp2 and bdccp3 genes. Using previously described sequences of bdccp1, bdccp2 and bdccp3, we have established a quantitative real-time PCR (qRT-PCR) assay for detection and assessment of the efficiency of B. divergens gametocytes production in bovine blood. We analysed fluctuations in expression of bdccp genes during cultivation in vitro, as well as in cultures treated with different drugs and stimuli.ResultsWe demonstrated that all B. divergens clonal lines tested, originally derived from naturally infected cows, exhibited sexual stages. Furthermore, sexual commitment was stimulated during continuous growth of the cultures, by addition of specific stress-inducing drugs or by alternating cultivation conditions. Expression of bdccp genes was greatly reduced or even lost after long-term cultivation, suggesting possible problems in the artificial infections of ticks in feeding assays in vitro.ConclusionsOur research provides insight into sexual development of B. divergens and may facilitate the development of transmission models in vitro, enabling a more detailed understanding of Babesia-tick interactions.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1731-y) contains supplementary material, which is available to authorized users.

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“…Recently, it was proposed that during the Babesia spp. sexual phase, some specific proteins with known functional roles in recognition and adhesion are expressed, including glycosylphosphatidylinositol (GPI) anchored proteins that interact with specific targets in the epithelial cells (Bastos et al, 2013 ; Alzan et al, 2016 ).…”

Section: Tick Midgut Molecules With a Role In Babesia mentioning

confidence: 99%

Deciphering Babesia-Vector Interactions

Antunes

Rosa

Couto

et al. 2017

Front. Cell. Infect. Microbiol.

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Understanding host-pathogen-tick interactions remains a vitally important issue that might be better understood by basic research focused on each of the dyad interplays. Pathogens gain access to either the vector or host during tick feeding when ticks are confronted with strong hemostatic, inflammatory and immune responses. A prominent example of this is the Babesia spp.—tick—vertebrate host relationship. Babesia spp. are intraerythrocytic apicomplexan organisms spread worldwide, with a complex life cycle. The presence of transovarial transmission in almost all the Babesia species is the main difference between their life cycle and that of other piroplasmida. With more than 100 species described so far, Babesia are the second most commonly found blood parasite of mammals after trypanosomes. The prevalence of Babesia spp. infection is increasing worldwide and is currently classified as an emerging zoonosis. Babesia microti and Babesia divergens are the most frequent etiological agents associated with human babesiosis in North America and Europe, respectively. Although the Babesia-tick system has been extensively researched, the currently available prophylactic and control methods are not efficient, and chemotherapeutic treatment is limited. Studying the molecular changes induced by the presence of Babesia in the vector will not only elucidate the strategies used by the protozoa to overcome mechanical and immune barriers, but will also contribute toward the discovery of important tick molecules that have a role in vector capacity. This review provides an overview of the identified molecules involved in Babesia-tick interactions, with an emphasis on the fundamentally important ones for pathogen acquisition and transmission.

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Differential Expression of Three Members of the Multidomain Adhesion CCp Family in Babesia bigemina, Babesia bovis and Theileria equi

Cited by 39 publications

References 38 publications

Targeted silencing of the Aquaporin 2 gene of Rhipicephalus (Boophilus) microplus reduces tick fitness

Targeted silencing of the Aquaporin 2 gene of Rhipicephalus (Boophilus) microplus reduces tick fitness

Stimulation and quantification of Babesia divergens gametocytogenesis

Deciphering Babesia-Vector Interactions

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