Fecal samples fail in PCR-based diagnosis of malaria parasite infection in birds
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Fecal samples fail in PCR-based diagnosis of malaria parasite infection in birds E. S. Martinsen • H. Brightman • R. C. Fleischer
Received: 27 August 2013 / Accepted: 11 August 2014 / Published online: 19 August 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Malaria parasites are common in wild vertebrates on all warm continents and have recently been isolated from wild apes by non-invasive fecal sampling. Here, we examined the utility of fecal samples for malaria parasite detection in wild birds. We collected both blood and fecal samples from 56 birds sampled in the field, extracted DNA from all samples using various methods, and screened all samples using sensitive PCR-based methods. We found 35 birds to be positive for malaria parasite infection (genera Plasmodium and Parahaemoproteus) using blood samples while no fecal samples revealed a positive infection. These results suggest that malaria parasites cannot be efficiently detected from fecal samples of birds and that blood sampling is still necessary for the study of the malaria parasites of wild bird populations. Keywords Avian malaria parasites Fecal diagnostics Haemosporidians The malaria parasites are an extremely successful and diverse group of blood parasites that infect mammals, squamate reptiles, crocodiles, turtles, and birds worldwide (Garnham 1966; Martinsen et al. 2008). Historically, these parasites have been studied by microscopy of blood films prepared from live vertebrate hosts. For elusive, rare, and sensitive vertebrate species, many of which are difficult to capture and sample in the field and for which sampling logistics including permitting prove difficult, parasite sampling has been limited. Recently, standard PCR-based
E. S. Martinsen (&) H. Brightman R. C. Fleischer Center for Conservation and Evolutionary Genetics, Smithsonian Conservation Biology Institute, National Zoological Park, P. O. Box 37012, MRC5503, Washington, DC 20013-7012, USA e-mail: [email protected]
methods have been used to detect parasite infection from non-traditional samples including archived blood and other tissue samples including those from extinct species or historical populations. The combination of PCR-based methods and non-invasive sampling techniques has allowed for parasite sampling from the great apes, a group under strict protection and for which we have limited information on their malaria parasite fauna. Using fecal samples collected in the field, a number of research groups have successfully genotyped malaria parasites of the genus Plasmodium from chimpanzees (Pan troglodytes), western gorillas (Gorilla gorilla), eastern gorillas (Gorilla beringei), and bonobos (Pan paniscus) (Kaiser et al. 2010; Liu et al. 2010; Prugnolle et al. 2011). Screening for malaria parasite infection from feces has also been successfully carried out in humans (Jirku et al. 2012). As birds are notoriously difficult to sample in the field, and in some cases blood sampling is not permitted or feasible, we evaluated for the
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