The use of molecular and morphological characters to resolve the taxonomic identity of cryptic species : the case of Miniopterus manavi (Chiroptera, Miniopteridae)

Based on recent molecular phylogenetic studies, the Old World bat family Miniopteridae, composed of species in the genus Miniopterus, has been shown to contain complex paraphyletic species, many of which are cryptic based on convergent morphological characters. Herein we resolve the phylogenetic relationships and taxonomy of the species complex M. manavi on Madagascar and in the Comoro Archipelago, where these animals occur in different bioclimatic zones. First using mitochondrial cytochrome-b sequence data to define clades and then morphology to corroborate the molecular data, including comparisons to type specimens, we demonstrate that animals identified as this taxon are a minimum of three species: M. manavi sensu stricto occurs in at least the central portion of the Central Highlands; M. griveaudi has a broad distribution in lowland northern and central western Madagascar and the Comoros (Anjouan and Grande Comore), and M. aelleni sp. n. has been found in northern and western Madagascar and the Comoros (Anjouan). In each case, these three clades were genetically divergent and monophyletic and the taxa are diagnosable based on different external and craniodental characters. One aspect that helped to define the systematics of this group was isolation of DNA from one of the paratypes of M. manavi collected in 1896 and new topotypic material. Miniopterus manavi is most closely allied to a recently described species, M. petersoni. At several localities, M. griveaudi and M. aelleni have been found in strict sympatry, and together with M. manavi sensu stricto show considerable convergence in morphological characters, but are not immediate sister taxa. In defining and resolving the systematics of cryptic species, such as miniopterid bats, the process of defining clades with molecular tools, segregating the specimens accordingly, and identifying corroborative morphological characters has been notably efficient.

The use of molecular phylogenetic and morphological tools to identify cryptic and paraphyletic species : examples from the diminutive long-fingered bats (Chiroptera : Miniopteridae : Miniopterus) on Madagascar

Based on nearly complete (1125 bp) cytochrome-b sequence data and morphological characters, two new endemic species of Miniopterus are described from Madagascar that were previously identified as M. manavi. Using phylogenetic analysis, the basal nodes of major lineages in the Malagasy members of this genus are weakly supported, while, in most cases, the branches leading to each of the clades are well resolved. Miniopterus mahafaliensis, new species, occurs in the southwestern semidesert areas and M. brachytragos, new species, has a broad distribution across the northern half of the island, ranging across several different biomes. Phylogenetic inference indicates that these two new taxa are not closely related to M. manavi sensu stricto, with average genetic distances of 9.2% and 5.7% from this taxon, respectively. On the basis of this and previous revisions, the former M. manavi complex is now recognized to represent at least five taxa, which do not form a monophyletic group with respect to one another, and represent extraordinary examples of convergent evolution. Miniopterus brachytragos is closely related to the recently named M. aelleni, while M. mahafaliensis is not closely associated with any of these species. Molecular phylogenetic analysis was imperative to resolve the species limits of these taxa and morphology then provided the means to corroborate the recovered clades. There are localities on the island, specifically limestone karstic zones, where four species of the former M. manavi sensu lato complex occur in strict sympatry. These species often use the same day-roost caves and have similar external and craniodental measurements. This raises intriguing questions as to how these animals divide their worlds with regard to dietary regimes and foraging strategies, as well as their speciation history.

Patterns of morphological and genetic variation in the endemic Malagasy bat Miniopterus gleni (Chiroptera: Miniopteridae), with the description of a new species, M. griffithsi

Over the past decade, major advances have been made concerning the systematics and species diversity of Malagasy bats, largely based on specimens collected during inventories and associated morphological and molecular genetic studies. Herein we describe a new species of endemic bat from southern Madagascar, Miniopterus griffithsi sp. n., which is the sister taxa to Miniopterus gleni, a taxon described in 1995 (holotype from Sarodrano, just north of the Onilahy River in the southwest). Based on current information, M. griffithsi is found in the sub-arid bioclimatic zone, south of the Onilahy River, and M. gleni occurs in a variety of different bioclimatic zones, north of the Onilahy River to the northern portion of the island and on the near shore island of Ile Sainte Marie. The realization that M. griffithsi was a separate entity was first based on phylogeographic studies of the M. gleni complex. Comparisons using 397 bp of mitochondrial cytochrome b found a divergence of 1.2% within animals occurring across much of Madagascar north of the Onilahy River, 0.07% in those south of the Onilahy River, and 7.4% in populations separated by this river. Subsequently, morphological characters were identified that supported the specific separation of populations occurring south (M. griffithsi) and north of the Onilahy River (M. gleni), which include tragus shape, pelage coloration, and skull proportions.

Exechonella (Exechonellidae: Bryozoa, Cheilostomata) from the recent and miocene of Southern Australia

The cheilostome family Exechonellidae Harmer, 1957 is widely distributed in time and space. The genus Exechonella Duvergier, 1924 has a pan-tropical to subtropical distribution from the Eocene to the Recent and is represented by several Australian species from the Tertiary of Victoria and the Recent ofthe southern and eastern coasts. Some species exhibit a wide range of variation in morphological characters, and one, Exechonella papillata, "appears to be new, and is described here. Nearly all specimens are encrusting, but one Tertiary Victorian species has erect, cylindrical, branching colonies. Recent samples, from a depth range of 40-190 metres, include large colonies of several thousand zooids. Frontal wall structures include marginal septular pores connecting between the visceral and hypostegal coeloms, and frontal foramina. Avicularia and homologous structures derived from frontal septular pores are illustrated. The structure of the frontal foramina in different populations of Tertiary E. marginata, demonstrates a major development of hypostegal coelom not found in other species, but resembling that found in another exechonellid genus, Stephanopora Kirkpatrick, 1888.

Does behavior reflect phylogeny in swiftlets (Aves: Apodidae)? A test using cytochrome b mitochondrial DNA sequences

Swiftlets are small insectivorous birds, many of which nest in caves and are known to echolocate. Due to a lack of distinguishing morphological characters, the taxonomy of swiftlets is primarily based on the presence or absence of echolocating ability, together with nest characters. To test the reliability of these behavioral characters, we constructed an independent phylogeny using cytochrome b mitochondrial DNA sequences from swiftlets and their relatives. This phylogeny is broadly consistent with the higher classification of swifts but does not support the monophyly of swiftlets. Echolocating swiftlets (Aerodramus) and the nonecholocating "giant swiftlet" (Hydrochous gigas) group together, but the remaining nonecholocating swiftlets belonging to Collocalia are not sister taxa to these swiftlets. While echolocation may be a synapomorphy of Aerodramus (perhaps secondarily lost in Hydrochous), no character of Aerodramus nests showed a statistically significant fit to the molecular phylogeny, indicating that nest characters are not phylogenetically reliable in this group.

Sympatric cryptic species in the crinoid genus Cenolia (Echinodermata: Crinoidea: Comasteridae) delineated by sequence and microsatellite markers

The marine species of the southern coast of Australia have not been well studied with regard to molecular connectivity. Cryptic species are expected to be prevalent on this coastline. Here, we investigate the crinoid genus Cenolia (Echinodermata: Crinoidea: Comasteridae) using molecular methods to elucidate cryptic species and phylogenetic relationships. The genus Cenolia dominates the southern Australian crinoid fauna in shallow waters. Few studies have examined crinoids for cryptic species at a molecular level and these have been predominantly based on mitochondrial data. We employ the nuclear markers 28S rRNA and ITS-2 in addition to the mitochondrial COI. Six divergent mitochondrial clades were identified. Gene flow between confirmed clades was subsequently examined by the use of six novel microsatellite markers, showing that sympatric taxa with low mtDNA divergences (1.7% K2P) were not interbreeding in the wild. The type specimens of Cenolia benhami and C. spanoschistum were examined, as well as all six divergent clades. Morphological characters dividing taxa were refined. Due to comb pinnule morphology, the New Zealand species benhami was determined to belong to the genus Oxycomanthus (nov. comb.). Three new species of Cenolia (including the Australian "benhami") require description. © 2014 Elsevier Inc.

A plant macrofossil identification tool for south-west Victoria

The ‘Seeds of South-Western Victoria’ database is presented here. The database is a digital collection of seed information that can be searched based on a range of seed morphological characters. Additionally it provides colour photographs of seeds and ecological detail of the plant species. It aims to facilitate plant macrofossil studies in Australia by aiding the identification of unknown sub-fossil seed material. It contains descriptions of 156 commonly occurring species of the Victorian Volcanic Plain, but design allows for continual addition of species from this region and elsewhere. The application of the database system to plant macrofossil identification, from both surface and fossil samples allows, for the first time, comparability of fossil assemblages and modern vegetation at a resolution of relevance to ecologists and conservationists. The taxonomic refinement of the pollen data provided by the macrofossils adds to a better understanding of recent vegetation dynamics and management, demonstrating the applicability of the technique to palaeoenvironmental analyses. A current application is to better understand the past environments and activities of Indigenous people on the Mt Eccles lava flow and assist in landscape restoration for potential World Heritage listing.

An integrative approach to characterize Malagasy bats of the subfamily Vespertilioninae Gray, 1821, with the description of a new species of Hypsugo

© 2015 The Linnean Society of London. Although important advances have been made in recent years in the taxonomy of different families and subfamilies of Malagasy bats, those belonging to the Vespertilioninae remain partially unresolved. Herein using a mitochondrial marker (cytochrome b) as the point of departure for 76 specimens of Malagasy vespers and appropriate African taxa, we diagnose the six taxa of this subfamily on the island by overlaying different morphological and bioacoustic characters on the clade structure of sequenced animals. The species include: endemic Neoromicia matroka, which is sister to African N. capensis; endemics N. malagasyensis and N. robertsi, which form sister species; a member of the genus Hypsugo, which is sister to African H. anchietae and described herein as new to science; Pipistrellus hesperidus for which Madagascar animals are genetically close but distinct from African populations of the same species; and endemic P. raceyi, which shows segregation of eastern (mesic) and western (dry) populations and its sister species relationships are unresolved. While the external and craniodental measurements, as well as bioacoustic variables, allow only partial differentiation of these six species of Vespertilioninae, molecular characters provide definitive separation of the taxa, as do male bacular morphology.

Antitropicality and convergent evolution: a case study of Permian neospiriferine brachiopods

Antitropical distribution is a biogeographical pattern characterized by natural occurrences of the same species or members of the same clade in the middle- or middle-to-high-latitudinal habitats of both hemispheres, either on land or in marine environments, without appearing in the intervening tropical environments. For most of the noted examples of Permian antitropical distribution, particularly in marine invertebrates, the causes of disjunctions have been mainly linked to either dispersal or vicariance models. Little attention has been paid to other possible mechanisms. This study investigated the antitropicality of some Permian neospiriferine brachiopods through detailed taxonomic revision, comparison of palaeobiogeographical distribution, and a phylogenetic analysis. Several species, previously assigned to Kaninospirifer, are here reassigned to other genera, especially to Fasciculatia in the northern hemisphere and to Quadrospira in the southern hemisphere during the Permian. Both Kaninospirifer and Fasciculatia appear to have been restricted to north-western Pangea and north-eastern Asia during the Permian, but there is no robust evidence to suggest their presence in the southern hemisphere to which Imperiospira and Quadrospira were confined. In spite of the distributional separation between the two pairs of neospiriferine genera in the Permian palaeobiogeographical regime, they share considerable numbers of morphological characters, such as a large shell, subdued fasciculation, and reduction of ventral adminicula. Notwithstanding these morphological similarities, our phylogenetic reconstruction of the neospiriferines does not support a close relationship between these genera. This therefore must indicate that these similar morphological features were independently acquired, probably with these taxa living in spatially separate but ecologically compatible environmental conditions in the mid-latitudinal area of each hemisphere during the Permian. We regard this as an example of convergent evolution.

Description of a new species of the miniopterus aelleni group (chiroptera: miniopteridae) from upland areas of central and northern Madagascar

Recent molecular genetic work, combined with morphological comparisons, on Malagasy members of the bat genus Miniopterus (Family Miniopteridae), has uncovered a number of cryptic species. Based on recently collected specimens and associated tissues, we examine patterns of variation in M. aelleni, the holotype of which comes from Ankarana in northern Madagascar. Using molecular genetic (mitochondrial cytochrome b) and morphological characters we describe a new species, M. ambohitrensis sp. nov. In northern Madagascar, M. ambohitrensis and M. aelleni are allopatric, but occur in relatively close geographical contact (approximately 40 km direct line distance) with M. ambohitrensis found at Montagne d'Ambre in montane humid forest and M. aelleni sensu stricto at Ankarana in dry deciduous forest. Morphologically, this new taxon is differentiated from M. aelleni based on pelage coloration, external measurements, craniodental differences, and tragus shape. Comparisons using 725 bp of cytochrome b found a divergence of 1.1% within M. aelleni sensu stricto, 0.8% within M. ambohitrensis, and 3.3% between these two clades. The two sister species do not demonstrate acoustical differences based on recordings made in a flight cage. Miniopterus ambohitrensis is known from four localities in the northern and central portions of Madagascar, all from montane regions and across an elevational range from about 800 to 1600 m; its calculated "Extent of occurrence" is 15,143 km². It is possible that this species is at least partially migratory.

Shedding new light on old species identifications : morphological and genetic evidence suggest a need for conservation status review of the critically endangered bat, Saccolaimus saccolaimus

Information based on the accurate identification of species is a vital component for achieving successful outcomes of biodiversity conservation and management. It is difficult to manage species that are poorly known or that are misidentified with other similar species. This is particularly problematic for rare and threatened species. Species that are listed under endangered species classification schemes need to be identified accurately and categorised correctly so that conservation efforts are appropriately allocated. In Australia, the emballonurid Saccolaimus saccolaimus is currently listed as ‘Critically Endangered’. On the basis of new observations and existing museum specimens, we used a combination of genetic (mitochondrial DNA sequence) and morphological (pelage characteristics, dig III : phalanx I length ratio, inter-upper canine distance) analyses to identify six new geographic records for S. saccolaimus, comprising ~100 individuals. Our analyses also suggested that there are likely to be more records in museum collections misidentified as S. flaviventris specimens. The external morphological similarities to S. flaviventris were addressed and genetic, morphological and echolocation analyses were used in an attempt to provide diagnostic characters that can be used to readily identify the two species in the field. We recommend genetic testing of all museum specimens of Australian Saccolaimus to clarify species’ distributions and provide data for reassessing the conservation status for both S. saccolaimus and S. flaviventris. Museum curators, taxonomists and wildlife managers need to be aware of potential species misidentifications, both in the field and laboratory. Misidentifications that result in misclassification of both threatened and non-threatened species can have significant implications.

Morphological, bioacoustical, and genetic variation in Miniopterus bats from eastern Madagascar, with the description of a new species

Recent molecular genetic work, combined with morphological comparisons, of Malagasy members of the bat genus Miniopterus (Family Miniopteridae), has revealed several cryptic species. Based on new specimens and associated tissues, we examine patterns of variation in the recently described species M. petersoni, the holotype of which comes from extreme southeastern Madagascar, and for which specimens from more northerly portions of eastern Madagascar were noted to show some morphological divergence from typical M. petersoni. On the basis of morphological and genetic (cytochrome b) characters we described a new species, M. egeri sp. nov. This taxon also shows bioacoustical differences from M. petersoni. Miniopterus egeri is widely distributed in the eastern portion of Madagascar across an elevational range from near sea level to 550 m. The specific status of moderately small Miniopterus from Montagne d'Ambre in the far north remains to be determined.

Phytophthora cinnamomi in native vegetation communities of southern Victoia - morphological variation and paragyny among isolates

Morphology has often been used as an indicator of variability within species. The present study investigated morphological and physiological characteristics of isolates of Phytophthora cinnamomi collected from diseased vegetation communities at Anglesea, Victoria, and isolates collected from other regions in the State. Characteristics studied included growth rate on potato-dextrose agar (PDA), corn-meal agar and V8-juice agar at 24°C, growth rate on V8 agar at 15°C, colony morphology on PDA, sporangial and gametangial morphology, sporangial production and mating type. Phenotypic variation was demonstrated in radial growth rate, colony morphology and sporangial dimensions. Sporangial and oogonial dimensions and sporangial production were not significantly different between isolates from different geographical regions. All isolates were found to be of the A2 mating type suggesting variation was derived asexually. Paragynal associations, in an organism characteristically defined as amphigynal, were observed following crossing with A1 isolates. This is the first such study undertaken in southern Victoria. The findings highlight the importance of appropriate management of an area of such high conservation value as the Anglesea Heath to contain the current infection and to prevent introduction of new isolates into the area.