A comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: Adeleorina) and its application for studying infection in natural populations
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Public Library of Science
2014-04-17
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| Online Access: | http://hdl.handle.net/10261/111263 http://dx.doi.org/10.13039/501100001871 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003381 |
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dig-ibe-es-10261-1112632021-12-28T16:01:49Z A comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: Adeleorina) and its application for studying infection in natural populations Maia, Joao P. M. C. Harris, David James Carranza, Salvador Gómez-Díaz, Elena Fundação para a Ciência e a Tecnologia (Portugal) European Commission Ministério da Educação e Ciência (Portugal) Ministerio de Educación y Ciencia (España) This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Identifying factors influencing infection patterns among hosts is critical for our understanding of the evolution and impact of parasitism in natural populations. However, the correct estimation of infection parameters depends on the performance of detection and quantification methods. In this study, we designed a quantitative PCR (qPCR) assay targeting the 18 S rRNA gene to estimate prevalence and intensity of Hepatozoon infection and compared its performance with microscopy and PCR. Using qPCR, we also compared various protocols that differ in the biological source and the extraction methods. Our results show that the qPCR approach on DNA extracted from blood samples, regardless of the extraction protocol, provided the most sensitive estimates of Hepatozoon infection parameters; while allowed us to differentiate between mixed infections of Adeleorinid (Hepatozoon) and Eimeriorinid (Schellackia and Lankesterella), based on the analysis of melting curves. We also show that tissue and saline methods can be used as low-cost alternatives in parasitological studies. The next step was to test our qPCR assay in a biological context, and for this purpose we investigated infection patterns between two sympatric lacertid species, which are naturally infected with apicomplexan hemoparasites, such as the genera Schellackia (Eimeriorina) and Hepatozoon (Adeleorina). From a biological standpoint, we found a positive correlation between Hepatozoon intensity of infection and host body size within each host species, being significantly higher in males, and higher in the smaller sized host species. These variations can be associated with a number of host intrinsic factors, like hormonal and immunological traits, that require further investigation. Our findings are relevant as they pinpoint the importance of accounting for methodological issues to better estimate infection in parasitological studies, and illustrate how between-host factors can influence parasite distributions in sympatric natural populations. © 2014 Maia et al. JPM was funded through a PhD grant (SFRH/BD/74305/2010) supported by a Fundação para a Ciência e a Tecnologia (FCT) doctoral fellowship under the Programa Operacional Potencial Humano – Quadro de Referência Estratégico Nacional funds (POPH-QREN) from the European Social Fund (ESF) and Portuguese Ministério da Educação e Ciência. EG-D was supported by a Juan de la Cierva contract from the Ministerio de Educación y Ciencia, Spain. Financial support was provided by project ERG-PARIS-276838 from the European Commission. DJH is supported by FEDER through the compete program, the project “Genomics and Evolutionary Biology” cofinanced by North Portugal Regional Operational Program (ON.2) under NSRF through the European Regional Development Fund. Peer Reviewed 2015-02-25T13:39:28Z 2015-02-25T13:39:28Z 2014-04-17 2015-02-25T13:39:28Z artículo http://purl.org/coar/resource_type/c_6501 doi: 10.1371/journal.pone.0095010 issn: 1932-6203 PLoS ONE 9(4): e95010 (2014) http://hdl.handle.net/10261/111263 10.1371/journal.pone.0095010 http://dx.doi.org/10.13039/501100001871 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003381 24743340 Publisher’s version http://dx.doi.org/10.1371/journal.pone.0095010 Sí open Public Library of Science |
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This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
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Fundação para a Ciência e a Tecnologia (Portugal) |
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Fundação para a Ciência e a Tecnologia (Portugal) Maia, Joao P. M. C. Harris, David James Carranza, Salvador Gómez-Díaz, Elena |
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Maia, Joao P. M. C. Harris, David James Carranza, Salvador Gómez-Díaz, Elena |
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Maia, Joao P. M. C. Harris, David James Carranza, Salvador Gómez-Díaz, Elena A comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: Adeleorina) and its application for studying infection in natural populations |
| author_sort |
Maia, Joao P. M. C. |
| title |
A comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: Adeleorina) and its application for studying infection in natural populations |
| title_short |
A comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: Adeleorina) and its application for studying infection in natural populations |
| title_full |
A comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: Adeleorina) and its application for studying infection in natural populations |
| title_fullStr |
A comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: Adeleorina) and its application for studying infection in natural populations |
| title_full_unstemmed |
A comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: Adeleorina) and its application for studying infection in natural populations |
| title_sort |
comparison of multiple methods for estimating parasitemia of hemogregarine hemoparasites (apicomplexa: adeleorina) and its application for studying infection in natural populations |
| publisher |
Public Library of Science |
| publishDate |
2014-04-17 |
| url |
http://hdl.handle.net/10261/111263 http://dx.doi.org/10.13039/501100001871 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003381 |
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