Speciation in the Genus Cloacina (Nematoda: Strongylida): Species flocks and Intra-Host Speciation

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Speciation in the Genus Cloacina (Nematoda: Strongylida): Species flocks and Intra-Host Speciation 1828 Speciation in the genus Cloacina (Nematoda: Strongylida): species flocks and intra-host speciation N. B. CHILTON1, M. A. SHUTTLEWORTH2, F. HUBY-CHILTON2,A.V.KOEHLER2, A. JABBAR2,R.B.GASSER2 and I. BEVERIDGE2* 1 Department of Biology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada 2 Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Parkville, Victoria, Australia (Received 1 April 2017; revised 13 June 2017; accepted 13 June 2017; first published online 12 July 2017) SUMMARY Sequences of the first and second internal transcribed spacers (ITS1 + ITS2) of nuclear ribosomal DNA were employed to determine whether the congeneric assemblages of species of the strongyloid nematode genus Cloacina, found in the forest- omachs of individual species of kangaroos and wallabies (Marsupialia: Macropodidae), considered to represent species flocks, were monophyletic. Nematode assemblages examined in the black-striped wallaby, Macropus (Notamacropus) dor- salis, the wallaroos, Macropus (Osphranter) antilopinus/robustus, rock wallabies, Petrogale spp., the quokka, Setonix bra- chyurus, and the swamp wallaby, Wallabia bicolor, were not monophyletic and appeared to have arisen by host colonization. However, a number of instances of within-host speciation were detected, suggesting that a variety of methods of speciation have contributed to the evolution of the complex assemblages of species present in this genus. Key words: Cloacina, Nematoda, Strongylida, speciation, species flocks, internal transcribed spacers. INTRODUCTION differences in the distribution of species within the gastro-intestinal tract (Ogbourne, 1976;Mfitilodze The phenomenon of ‘species flocks’, that is the and Hutchinson, 1985; Bucknell et al. 1995; occurrence of numerous species of congeneric (or Stancampiano et al. 2010). Comparably detailed confamilial) parasites in the same host species, has studies on the strongyloid nematodes of elephants been the focus of a number of studies, particularly and rhinoceroces have not been conducted, but of parasitic nematodes. One of the best-known four genera and 49 species belonging to the related examples of the phenomenon is that of the oxyuroid strongyloid tribes Kiluluminea, Murshidinea and nematodes found in the colon of tortoises (Schad, Quiloninea are known to occur in their large intes- 1963; Petter, 1966). In this instance, each species tines (Chabaud, 1957; Round, 1968; Canaris and of tortoise harbours large numbers of congeneric or Gardner, 2003; Beveridge and Jabbar, 2013). In confamilial nematode species, all belonging to the the case of kangaroos and wallabies (family family Pharyngodonidae (Petter, 1966). However, Macropodidae) some 36 genera and 256 species of there are differences in the longitudinal and radial nematodes belonging to the sub-family Cloacininae distributions of these nematode species within the occur in the sacculated forestomach (Beveridge and colon of the host (Schad, 1963). Other examples of Chilton, 2001), frequently in large numbers species flocks of parasitic nematodes are those (Beveridge and Arundel, 1979), with again, some found in the large intestines of equids, elephants degree of spatial separation within the stomach and rhinoceroses, as well as those in the sacculated (Mykytowycz, 1964; Arundel et al. 1979; Pamment forestomachs of kangaroos and wallabies (Inglis, et al. 1994). 1971). All of these nematodes have direct life- In a critical review of the phenomenon of species cycles with the ingestion of eggs or third-stage flocks in parasitic helminths, Kennedy and Bush larvae from the environment (Anderson, 2000). (1992) indicated several difficulties in the application In the case of equids (horses, donkeys and zebras), of this appellation to the examples cited above. First 14 genera and 50 species of strongyloid nematodes of all, these authors noted that species flocks, accord- belonging to the tribe Cyathostominea are currently ing to classical definitions, could be defined either by recognized (Lichtenfels et al. 2008), with the ecological parameters such as co-occurrence (Mayr, common co-occurrence of many species (Bucknell 1984) and endemism (Ribbink, 1984) or could be et al. 1996; Anjos and Rodrigues, 2003;Buet al. circumscribed phylogenetically, with a species flock 2009; Kuzmina et al. 2009), but again with being a monophyletic assemblage (Greenwood, 1984). In the former case, such associations of mul- * Corresponding author: Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, tiple congeners could develop through a number of Parkville, Victoria, Australia. E-mail: ibeve@unimelb. host colonization events, while in the latter case, edu.au the communities could evolve through intra-host Parasitology (2017), 144, 1828–1840. © Cambridge University Press 2017 doi:10.1017/S0031182017001238 Downloaded from https://www.cambridge.org/core. University of Athens, on 30 Sep 2021 at 12:41:00, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0031182017001238 Nematode species flocks in kangaroos 1829 speciation. Intra-host speciation (Price, 1980; of host specificity (Beveridge et al. 2002). In add- Poulin, 2007) or even sympatric speciation (Kunz, ition, the study of a single genus (although the 2002) are considered to be potentially common monophyly of Cloacina has not yet been investigated modes of evolution in parasites. using molecular methods) overcomes the difficulty The phylogenetic definition of species flocks has of deciding whether species flocks should be consid- been successfully applied to cichlid fishes ered as being composed of congeners or whether the (Seehausen, 2006)andtorockfishes (Alesandrini concept should expand to con-sub- familiar or con- and Bernardi, 1999), but the paucity of rigorous familiar taxa (see Kennedy and Bush, 1992), a poten- phylogenetic studies of parasitic nematodes means tial complication in the studies of the nematode that this definition is often not applicable. In the assemblages of equids. Furthermore, studies to case of the cyathostomin nematodes of equids (Equus date of different species of kangaroos and wallabies spp.), available molecular evidence suggests that they (seven species of the currently recognized 54 host do indeed form a monophyletic assemblage (Hung species were studied by Beveridge et al. 2002) have et al. 2000;McDonnellet al. 2000). The same situ- shown that assemblages of nematode species range ation may apply to the strongyloid nematodes of ele- across a continuum, from three to 12. This spectrum phants as most species belong to two related tribes, of hosts with varying numbers of co-occurring con- the Murshidiinea and Quiloninea (Lichtenfels, geners is not represented in any other host-parasite 1980). This may well also apply to the Cloacininae system as there remain only five extant species of in macropodids. However, appropriate morphological equids (all with a very similar parasite fauna), three and molecular phylogenetic studies are lacking. A species of elephants and four species of rhinoceros, similar situation pertains, in the case of molecular all relicts of formerly more diverse faunas studies, to the oxyuroid nematodes in tortoises for (Franzen, 2010). In addition a number of the latter which molecular data are lacking. As a consequence, host species are endangered thereby imposing a limi- among nematodes, it appears that the phylogenetic tation on parasitological studies. definition of a species flock may only be applicable In an attempt to address the question of whether currently to the cyathostomin nematodes of equids. the species flocks of Cloacina seen in macropodids A molecular study of species of Onchocerca occurring are monophyletic, Beveridge et al. (2002) undertook in cattle also suggests within-host speciation in this a phylogenetic analysis based on morphological nematode genus, but the result is dependent upon characters. However, given that only a limited the inclusion of remaining congeners (Morales-Hojas number of morphological characters was available, et al. 2006). a common phenomenon for parasitic nematodes, In considering the ecological definition of a the resulting phylogeny exhibited a relatively low species flock, Kennedy and Bush (1992) pointed consistency index. In spite of this reservation, it out that the criterion of ‘co-occurrence’ was also was suggested that while several nematode species difficult to apply since, for several parasite groups, pairs could be identified in individual host species, relevant data were not available on the co-occurrence there was no strong evidence for the existence of of congeneric or confamilial parasites within an indi- monophyletic species flocks (Beveridge et al. 2002). vidual host; rather published data described the It was therefore tentatively suggested that the assem- meta-community. This ecological difference is also blages might have arisen through host switching. reflected in phylogenetic studies in which the criter- Given the difficulties encountered in the use of ion of sympatry in potential examples of intra-host morphological characters to establish phylogenetic speciation can be problematical (McCoy, 2003). relationships among nematodes, it was decided to Data on the co-occurrence of species of nematodes re-examine the problem using molecular methods. are however available for the cyathostomins of The combined first and second internal transcribed equids (Bucknell et al. 1996; Anjos and Rodrigues, spacers (ITS-1 and ITS-2) of nuclear ribosomal
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