Coevolutionary Patterns and Diversification of Avian Malaria Parasites in African Sunbirds (Family Nectariniidae)
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1 Coevolutionary patterns and diversification of avian malaria parasites in African sunbirds (Family Nectariniidae) ELVIN J. LAURON1*, CLAIRE LOISEAU1, RAURI C. K. BOWIE2, GREG S. SPICER1, THOMAS B. SMITH3,MARTIMMELO4 and RAVINDER N. M. SEHGAL1 1 Department of Biology, San Francisco State University, San Francisco, California 94132, USA 2 Museum of Vertebrate Zoology and Department of Integrative Biology, University of California at Berkeley, Berkeley, California 94720, USA 3 Center for Tropical Research, University of California at Los Angeles, Los Angeles, California 90095, USA 4 CIBIO, Centro de Investigação em Biodiversidade e Recursos Genéticos, InBio, Universidade do Porto, Campus Agrário de Vairão, 4485-661 Vairão, Portugal (Received 14 July 2014; revised 15 September 2014; accepted 2 October 2014) SUMMARY The coevolutionary relationships between avian malaria parasites and their hosts influence the host specificity, geographical distribution and pathogenicity of these parasites. However, to understand fine scale coevolutionary host–parasite relation- ships, robust and widespread sampling from closely related hosts is needed. We thus sought to explore the coevolutionary history of avian Plasmodium and the widespread African sunbirds, family Nectariniidae. These birds are distributed throughout Africa and occupy a variety of habitats. Considering the role that habitat plays in influencing host-specificity and the role that host-specificity plays in coevolutionary relationships, African sunbirds provide an exceptional model system to study the processes that govern the distribution and diversity of avian malaria. Here we evaluated the coevolutionary histories using a multi-gene phylogeny for Nectariniidae and avian Plasmodium found in Nectariniidae. We then assessed the host–parasite biogeography and the structuring of parasite assemblages. We recovered Plasmodium lineages concurrently in East, West, South and Island regions of Africa. However, several Plasmodium lineages were recovered exclusively within one respective region, despite being found in widely distributed hosts. In addition, we inferred the biogeographic history of these parasites and provide evidence supporting a model of biotic diversification in avian Plasmodium of African sunbirds. Key words: Plasmodium, nectariniidae, cospeciation, African sunbird, avian malaria, host switching. INTRODUCTION molecular approaches allows researchers to elucidate the phenomena of cospeciation, the hallmark of co- Coevolution and natural selection may shape the evolution (Demastes and Hafner, 1993; Clark et al. interactions among species by presenting an evol- 2000). For example, studies of mites and their utionary trade-off between specializing to perform a avian hosts show significant evidence of cospeciation few activities well, and generalizing to perform many (Ehrnsberger, 2001; Morelli and Spicer, 2007; activities fairly (Levins, 1968; Brodie et al. 1999). Hendricks et al. 2013). In addition, mammalian and Thus, the ability for parasites to specialize or gen- avian hosts also show evidence of cospeciation with eralize and to infect hosts with varying efficacy poses haemosporidian blood parasites, including the causa- compelling questions: what factors determine para- tive agents of leucocytozoonosis and malaria (Ricklefs site host-specificity, and how does the degree of and Fallon, 2002; Garamszegi, 2009; Hughes and host-specificity impact parasite distribution and di- Verra, 2010; Jenkins and Owens, 2011). versification? These factors may ultimately influence Infections of malaria occur in multiple vertebrate the virulence, the geographical distributions, and the hosts including reptiles, mammals and birds (Levine, potential for parasites to emerge into novel hosts 1988). Malaria parasites of birds are found on all con- (Garamszegi, 2006; Hellgren et al. 2009; Cooper et al. tinents of the world, except Antarctica (Valkiunas, 2012). 2005), and have been studied intensively for over 100 The host range of parasites primarily depends years. Avian malaria has therefore proven to be an on their compatibility with the host, which is limited important model system for pursuing evolutionary by a coevolutionary arms race between hosts and and ecological issues such as speciation (Ricklefs parasites (Kawecki, 1998). In fact, the study of and Fallon, 2002; Ricklefs et al. 2004; Pérez-Tris host–parasite coevolutionary relationships through et al. 2007), life-history trade-offs (Garamszegi, 2006; Hellgren et al. 2009), and competition and com- * Corresponding author. Department of Biology San Francisco State University, 1600 Holloway Avenue, munity structure in a changing environment (Read San Francisco, California 94132, USA. E-mail: elauron@ and Taylor, 2001; Mideo et al. 2008; Mideo, 2009; mail.sfsu.edu Palinauskas et al. 2011; Loiseau et al. 2012; Parasitology, Page 1 of 13. © Cambridge University Press 2014 doi:10.1017/S0031182014001681 Elvin J. Lauron and others 2 Svensson-Coelho et al. 2013). Extensive host switch- Plasmodium vector distribution, which may conse- ing of some avian Plasmodium lineages has been quently affect the transmission and prevalence of shown in birds belonging to different families avian malaria (Kamdem et al. 2012; Pavlacky et al. (Ricklefs and Fallon, 2002; Waldenström et al. 2012). Because avian Plasmodium are globally dis- 2002; Fallon et al. 2003, 2005; Szymanski and tributed among many bird species, the coevolution of Lovette, 2005) and the potential for these widely avian Plasmodium parasites are likely driven at the distributed avian Plasmodium parasites to switch into global-scale by environmental changes with periodic new hosts can have calamitous effects, as was seen in cycling of geographic expansion and isolation, a the endemic bird populations of Hawaii (van Riper, process described as taxon pulses (Hoberg and 1986). Thus, it is crucial to understand the factors Brooks, 2008; Agosta et al. 2010). Here we assess that influence parasite host specificity and the the cross-continental patterns and coevolutionary longstanding host relationships among parasites. histories of a host–parasite system in Africa and Considering host–parasite relationships, Ricklefs provide empirical evidence that supports a model for and Fallon (2002) used tree-based methods to assess biotic diversification in avian Plasmodium. cospeciation and host switching between the avian haemosporidian genera Plasmodium and Haemo- MATERIALS AND METHODS proteus and their hosts. Their study included 20 avian families distributed throughout six countries, Sample collection in which they reported significant cospeciation and Blood samples from birds were collected in 12 host switching. In addition, Ricklefs et al.(2004) countries of Africa and in three island countries conducted a more focused analysis by re-examining covering five biogeographic regions: West Africa cospeciation of avian malaria parasites and forest (Gabon, Ghana, and Cameroon; N = 935), East dwelling songbirds within North America and the Africa (Burundi, Kenya, Democratic Republic of West Indies. Consistent with their previous findings, the Congo, Malawi, Mozambique, Rwanda, Uganda, cospeciation and host switching were prominent Tanzania and Zambia; N = 392), Southern Africa events recovered in their study. Here, we assessed (The Republic of South Africa; N = 252), the coevolutionary host–parasite relationships of Madagascar (N = 18) and two small oceanic archipe- avian Plasmodium found in the Old-World bird lagos (São Tomé and Principe; N = 21, and the family Nectariniidae. We sought to increase the sen- Comoros; N = 18) during the period 1999 to 2009. sitivity for the detection of coevolutionary relation- All birds were caught with mist-nets and blood ships by focusing on the large assemblage of Africa samples were collected from the brachial vein. Blood sunbirds. This assemblage comprises 95 species that samples were stored in lysis buffer (10 mM Tris-HCL are primarily restricted to continental Africa and its pH 8·0, 100 mM EDTA, 2% SDS) or in absolute neighbouring islands including Madagascar, but ethanol at −80 °C or in liquid Nitrogen cryo-tanks. which includes five Asian species that represent a recent back-colonization from Africa (Bowie, 2003, PCR amplification and DNA sequencing Bowie et al. unpublished data). Sunbirds utilize virtually all forms of habitat, In total, 1636 individual sunbirds (N species = 32) including moist forests, swamps, grasslands, primary were screened for Plasmodium parasites (see Table 1). rainforests, open woodlands, and semi-arid regions DNA was extracted from whole blood following a (Cheke et al. 2001). Most species are sedentary or DNeasy kit protocol (Qiagen, Valencia, California). short-distance seasonal migrants, and can be either Success of each DNA extraction was verified with ecological generalists or specialists (i.e. restricted primers that amplify the brain-derived neurotrophic to a very small area, e.g. highlands in East Africa; factor (BDNF) (Sehgal and Lovette, 2003). Bowie et al. 2004). A recent study established that Since mitochondrial lineages have been shown to be Nectariniidae and other bird families in Africa reproductively isolated (Bensch et al. 2004), Plasmo- (Muscicapidae, Pycnonotidae, Timaliidae and dium spp. mitochondrial haplotypes are often defined Turdidae) also harbour both generalist and specialist as unique cytochrome b lineages (Hellgren et al. 2004; Plasmodium parasites (Loiseau et al. 2012). The Waldenström et al. 2004; Bensch et al. 2009). There- detection of