Putative Sister Species Research Articles

Spiranthes hachijoensis (Orchidaceae), a new species within the S. sinensis species complex in Japan, based on morphological, phylogenetic, and ecological evidence

The systematics of the Old World Spiranthes sinensis (Pers.) Ames species complex (Orchidaceae) has been complicated by its wide distribution and morphological variations. Within the species complex, S. australis Lindl. has been generally accepted as the only Spiranthes Rich. species distributed on the Japanese mainland. The present study provides morphological, phylogenetic, and ecological evidence for the recognition of S. hachijoensis Suetsugu as a new species of the S. sinensis species complex on the Japanese mainland. Spiranthes hachijoensis is morphologically similar to S. hongkongensis S.Y. Hu & Barretto and S. nivea T.P. Lin & W.M. Lin, sharing a degenerated rostellum, pollinia without a viscidium, and distinctly trilobed stigma. However, the taxon can be morphologically distinguished from S. hongkongensis by its glabrous rachis, ovaries, and sepals, and from S. nivea by its papillate labellum disc, larger papillate basal labellum callosities, and glabrous rachis, ovaries, and sepals. The autogamy and flowering phenology (i.e., earlier flowering) of S. hachijoensis are most likely responsible for premating isolation from the sympatric S. australis. A MIG-seq-based high-throughput molecular analysis indicated that the genetic difference between S. hachijoensis and its putative sister species S. sinensis is comparable to, or even greater than, the genetic difference between pairs of other species within the S. sinensis species complex. Our multifaceted approach strongly supports the recognition of S. hachijoensis as a morphologically, phenologically, phylogenetically, and ecologically distinct species.

Neoponera Emery, 1901 (Hymenoptera: Formicidae) revisited: 1. The N. laevigata species-group

Abstract The Neotropical Neoponera laevigata species-group is revised. This group can be separated from others in the genus by means of a new key to Neoponera species–groups. We propose two new species, N. gojira sp. nov., and N. mashpi sp. nov., the latter being usually confused with its putative sister species, N. laevigata (Smith). This small, termite-feeding species-group now contains five taxa. We provide comprehensive redescriptions for the known castes of the three previously known species, together with images and notes about intraspecific morphological variation, biology, and distribution of the taxa here treated. We reveal the external morphological features of the genitalia of the known males. This is the first contribution en route to a better understanding of the systematics of this charismatic ponerine genus.

World Heritage lizard: population genetics and species status of the range-restricted Hamelin skink, Ctenotus zastictus

The Shark Bay World Heritage region in western Australia is home to a number of species of substantial conservation concern. Among these is a small scincid lizard, Ctenotus zastictus, which represents one of the most geographically-restricted vertebrates on the Australian mainland. The long-term persistence of Ctenotus zastictus is threatened due to the small size of its range, isolation from suitable habitat patches elsewhere, and potential impacts from climate change and mining. Accordingly, conservation efforts in Australia have targeted C. zastictus as the focus of protection. But this attention might be unwarranted – the species might not be evolutionarily unique. Previous genetic assessments have suggested limited differentiation between C. zastictus and its putative sister taxon, and the taxonomic status of C. zastictus has never been formally evaluated. Here, we use population genomic, phylogenetic, and ecoclimatic analyses to characterize the species status of C. zastictus in context of its closely-related congeners. In doing so, we explore the practical and conceptual challenges of revising species boundaries in threatened species, many of which are also rare and range-restricted. We demonstrate that C. zastictus is a coherent evolutionary unit that has been isolated from its putative sister species for at least two million years. Based on these results, we recommend that C. zastictus should retain its taxonomic status.

Integrative taxonomy supports two new species of Chimarra Stephens, 1829 from Brazil (Trichoptera: Philopotamidae)

Chimarra Stephens, 1829 is the largest genus of the Philopotamidae with about 930 species and cosmopolitan distribution. Recent taxonomic revisions have subdivided the genus into four subgenera: Chimarra, Curgia Walker, 1860, Chimarrita Blahnik, 1997, and Otarrha Blahnik, 2002, the last three restricted to the New World. In this paper, we describe and illustrate two new species of Chimarra from Brazil, C. (Otarrha) paraodontasp. nov. from Rio de Janeiro State and C. (Chimarrita) truncatasp. nov. from Amazonas and Pará states. Partial sequences of cytochrome oxidase subunit I (COI, DNA barcodes) were generated and integrated with morphological evidence to delimit the new species and evaluate their phylogenetic relationships within the genus. A maximum likelihood analysis of 48 COI sequences representing 19 species of Chimarra corroborated their subgeneric assignment based on morphology and highlighted their putative sister species. Both new species showed high K2P divergences when compared to their sister species: Chimarra (O.) paraodontasp. nov. and C. (O.) odonta (17.4–21.3%) and Chimarra (C.) truncatasp. nov. and C. (C.) simpliciforma (20.0–21.3%). These distances are comparable to the range of interspecific distances calculated for the whole genus (13.6–22.7%), adding support to their description as new species. This analysis was especially important because of the high morphological similarity of C. paraodontasp. nov. and C. odonta. Finally, analysis of the sequences of Chimarra odonta suggests that the nominal species may represent a complex of cryptic species with high intraspecific divergences (up to 18.1%), with at least two of those lineages co-occurring with C. paraodontasp. nov. at Parque Nacional do Itatiaia.

Mitochondrial genomes of the freshwater monogonont rotifer Brachionus fernandoi and of two additional B. calyciflorus sensu stricto lineages from Germany and the USA (Rotifera, Brachionidae)

The Brachionus calyciflorus species complex was recently subdivided into four species, but genetic resources to resolve phylogenetic relationships within this complex are still lacking. We provide two complete mitochondrial (mt) genomes from B. calyciflorus sensu stricto (Germany, USA) and the mt coding sequences (cds) from a German B. fernandoi. Phylogenetic analysis placed our B. calyciflorus sensu stricto strains close to the published genomes of B. calyciflorus, forming the putative sister species to B. fernandoi. Global representatives of B. calyciflorus sensu stricto (i.e. Europe, USA, and China) are genetically closer related to each other than to B. fernandoi (average pairwise nucleotide diversity 0.079 intraspecific vs. 0.254 interspecific).

Ecological niche divergence between the brown recluse spiders Loxosceles laeta and L. surca (Sicariidae) in Chile

ABSTRACT The laeta species group is the main source of Loxosceles species in Chile, a genus of medical importance due to the cytotoxic properties of its venom and the synanthropic habits of its dominant species, Loxosceles laeta. However, a poorly studied, cryptic, and putative sister species, Loxosceles surca, is also known to live in Chile at latitudes that overlap with L. laeta. In contrast to L. laeta, L. surca lives in natural areas away from humans. We used DNA sequences from L. surca to infer the phylogenetic history of this species and its congenerics. Additionally, we used ecological niche modelling to define the ecology of this species in contrast to its sister species. Our results show that L. surca does, indeed, belong to the laeta species group, and is likely a sister species to L. laeta. Although these species are fairly cryptic with respect to each other, the substantial genetic divergence between them is consistent with other distinct sister species in this genus. The laeta group is also sister to the large reclusa group that radiated into the Caribbean, Central, and North America. The phylogeny further supports the contention that L. diaguita belongs in its own species group separate from laeta. Our ecological niche analysis shows that L. surca and L. laeta have distinctly different allopatric habitats, and we suggest that these differences explain why one species became synanthropic, while the other did not.

Regression of dark color in subterranean fishes involves multiple mechanisms: response to hormones and neurotransmitters

ABSTRACT Organisms with source-populations restricted to the subterranean biotope (troglobites) are excellent models for comparative evolutionary studies, due to their specialization to permanent absence of light. Eye and dark pigment regression are characteristics of most troglobites. In spite of the advance in knowledge on the mechanisms behind eye regression in cave fishes, very little is known about pigmentation changes. Studies were focused on three species of the genus Pimelodella. Exemplars of the troglobitic P. spelaea and P. kronei were compared with the epigean (surface) P. transitoria, putative sister-species of the latter. Melanophore areas and densities are significantly lower in the troglobitic species. Evaluating the in vitro response of these cells to adrenaline, acetylcholine and MCH, we observed a reduced response in both troglobites to adrenaline. The same trend was observed with MCH, but not statistically significant. No response to acetilcholine was detected in all the three. Contrary to expectations, even though eye-regression in P. spelaea was much lower than in P. kronei, pigmentation regression was more advanced. Multiple mechanisms of loss showing a mosaic of traits in troglobitic fishes are discussed here.

Historical biogeography of fishes from coastal basins of Maranhão State, northeastern Brazil

ABSTRACT The Amazonian ichthyofauna is one of the most diverse in the world, yet fishes from the adjacent coastal basins of Maranhão State in Northeastern Brazil remain poorly known. We use phylogeographic, community phylogenetic and phylogenetic beta diversity methods to study the biogeographic history of fishes from the coastal basins of Maranhão State. We report a total of 160 fish species from the basins of the Maranhão region, representing a 93% increase over results of previous studies. All the fish species assemblages from Maranhão are polyphyletic, with only a few putative sister species pairs inhabiting the region. The modern watershed divides among Maranhão basins do not form substantial barriers to dispersal for freshwater fish species, and are more effectively modelled as biogeographic islands than as biogeographic provinces. In combination these results suggest that the Maranhão ichthyofauna was assembled under the influence of several macroevolutionary (extinction, dispersal) and landscape evolution processes, during the Miocene and Pliocene, as well as by the modern ecological characteristics of the region. The results indicate that the distinctive geological and climatic conditions and history of Northeastern Brazil strongly constrained the formation of aquatic faunas in coastal basins of Maranhão State.

Ultrastructural and elemental characterization of the extracorporeal tube of the sessile rotifer Floscularia conifera (Rotifera: Gnesiotrocha)

Rotifers are aquatic microinvertebrates that live in the plankton or in the benthos, which may include a variety of macrophytes. Among these periphytic forms of rotifers, some have taken up a sessile existence and secrete protective tubes around their bodies. One type of tube common to species of Floscularia is made of small round pellets. To date, the building process and some fine structural details are known for Floscularia ringens, but many questions about the composition of the tube and its ultrastructure still remain unanswered. Here, we use transmission electron microscopy and scanning electron microscopy–energy dispersive X-ray spectroscopy (SEM–EDS) to study the ultrastructure of the pellets and their elemental composition, respectively, in the putative sister species, Floscularia conifera. We revealed several new details that add important information about the physiology of tube-making in species of Floscularia. First, we note an inner secretory membrane that is thin, electron lucent, and supports the external pellets. The pellets are relatively consistent in size and have a small depression on their inner surface. All pellets are individually wrapped in a secretory membrane that completely encapsulates suspended materials collected from the surrounding water. Elemental signatures of pellets reveal they consist mostly of carbon (C), nitrogen (N), and oxygen (O), with some silicon (Si) content that is likely the result of diatom shells. Other trace elements such as iron (Fe) and sodium (Na) are also present and likely the result of incorporated bacteria and suspended materials. When larval rotifers are cultured in filtered pond water, the pellets consist mostly of C and O, with little N and no Si; Fe is present in smaller amounts. These new discoveries provide a better understanding of the physiology of rotifer tube construction and tube composition, and their future utility in understanding if and how changes in freshwater environments might impact these factors.

Rediscovery of Dixa repanda Peters and description of its putative sister species, Dixa falcata sp. nov., from the south-central United States (Diptera: Dixidae).

Dixa repanda Peters, previously known only from the type series of three males, was rediscovered after targeted collections in the southcentral and midwestern United States. These collections also revealed the presence of its undescribed putative sister species, Dixa falcata sp. nov. The two species are sympatric in the Ozark Mountains. Adults of D. repanda are redescribed and those of Dixa falcata are described. Males of both species are distinguished from related, similarly colored species by a falcate gonostylus and extremely reduced cercus. Males of the two species are easily separated from each other by the shape and thickness of the gonostylus. Females are not reliably separable from each other or related species. Biological insights, including known distributions, of both species are provided.

Intrageneric chloroplast genome comparison in the genus Euglena (Phylum: Euglenophyta) with annotated chloroplast genomes of Euglena hiemalis and Euglena clara

The genus Euglena is composed of six subclades with a diversity of chloroplast morphologies, unlike that found in the other genera of Euglenaceae. This genus contains five published chloroplast genomes (cpGenome) and the colorless plastid genome (ptGenome) of the non-photosynthetic euglenoid Euglena longa within the same subclade as Euglena gracilis. Previous studies of cpGenomes revealed that Euglena viridis and E. gracilis, although in separate subclades, had few cpGenome differences, while Euglena mutabilis maintained the same gene order but was mirror-inverted except for the rRNA cluster. However, we expanded the number of cpGenomes available in Euglena by sequencing and annotating the cpGenomes of Euglena clara, the earliest diverging species in the E. gracilis and E. longa subclade and Euglena hiemalis, the putative sister species to E. longa. Analysis of these newly annotated cpGenomes showed them to be largely similar in gene content, conserved gene clusters (operons), and G + C/A + T percentage to previously published Euglena cpGenomes. The only extensive gene rearrangements observed were between cpGenomes and the ptGenome in subclade B. However, a unique feature of the subclade was multi-copies of the rRNA operon. Also, homologous twintrons in psbD and psbF to E. gracilis were observed in E. hiemalis. Overall, these results revealed a conserved intrageneric cpGenome for Euglena species.

Floral morphology and anatomy of Ophiocaryon, a paedomorphic genus of Sabiaceae.

Ophiocaryon is a lesser known genus in Sabiaceae. This study examines flowers of six Ophiocaryon species in comparison with Meliosmaalba, to identify taxonomically informative characters for understanding relationships within the family Sabiaceae, to imply previously unknown pollination mechanisms of Ophiocaryon, and to contribute to the placement of Sabiaceae within the early-diverging eudicots. Floral morphology and anatomy of six Ophiocaryon species and M. alba were studied and described using scanning electron microscopy, clearing techniques and resin sectioning. Novel characters of Ophiocaryon were identified, e.g. conical cells on petals, different kinds of orbicules in anthers, stomata on nectary appendage tips and ovary, two distinct surface patterns on stamens and ovary, tanniferous cell layers in the ovary wall, and acorn-shaped unitegmic ovules with very short integuments. Comparison of floral characters between Ophiocaryon and Meliosma found that the calyx, corolla, androecium and gynoecium of Ophiocaryon resemble an undeveloped state of the latter taxon, reflecting a paedomorphic regression of the flower of Ophiocaryon. The flower morphology and anatomy of Ophiocaryon was compared with its putative sister species M. alba, but no clear shared derived characters could be detected. Moreover, the findings of scent, presence of conical cells on petals and a nectary suggest flowers are pollinated by small insects with a secondary pollen presentation on the cupula of fertile stamens. We found that Ophiocaryon may be derived from ancestors that were similar to extant Meliosma in their flower structure and pollination mechanism. However, the lack of shared derived characters between Ophiocaryon and its phylogenetic sister group M. alba is puzzling and requires further investigations on the diversity of the latter species.

The systematics and biogeography of African tailorbirds (Cisticolidae: Artisornis) with comment on the choice of Bayesian branch-length prior when analyzing heterogeneous data

The Long-billed Tailorbird (Artisornis moreaui), one of Africa’s rarest birds, has a strikingly disjunct distribution, the origin of which has long puzzled biogeographers. One small population (subspecies moreaui) occurs in sub-montane forest in the East Usambara Mountains, a sky island near the coast of northern Tanzania, and another (subspecies sousae) on Serra Jeci in northwestern Mozambique, 950km away. The African Tailorbird, the putative sister-species of Long-billed Tailorbird, also occurs in the East Usambara Mountains and on Serra Jeci, but in addition occupies all the Eastern Arc Mountain forests between these disjunct sites. Stuart (1981) hypothesized that the two tailorbird distributions could be explained by strong ecological competition, with African Tailorbird populations having eliminated Long-billed Tailorbird populations via competitive exclusion in montane forests between the East Usambara and Serra Jeci. If such competitive exclusion explains these geographic distributions, the co-occurrence of the two species in the East Usambara and at Serra Jeci may be ephemeral, with the status of Long-billed Tailorbird especially in doubt. We sought to (1) determine whether the two species of African tailorbirds are indeed sister-species, and (2) test predictions from Stuart’s (1981) competitive exclusion hypothesis using genetic data. Phylogenetic analyses of our seven gene dataset (3 mtDNA, 4 introns; 4784bp) indeed place these two species together in the genus Artisornis. Instead of finding shallow divergence among African Tailorbird populations and deep divergence between Long-billed Tailorbird populations as expected from Stuart’s hypothesis, we recover deep genetic divergence and geographic structure among populations of both tailorbird species. This result is consistent with long-term co-existence of the two species at East Usambara and Serra Jeci. Observational data from both the East Usambara and Serra Jeci suggest that the two species have diverged in use of forest canopy strata. From a conservation standpoint, our results suggest that extinction of the Long-billed Tailorbird as a function of competition with African Tailorbird is highly unlikely, and should not be viewed as imminent. Threats to its survival are instead anthropogenic, and conservation measures should take this into account. Finally, our empirical results suggest that mis-specification of the branch-length prior in Bayesian analyses of mitochondrial DNA data can have a profound effect on the overall tree-length (sum of branch-lengths), whereas the topology and support values tend to remain more stable. In contrast, mis-specification of the branch-length prior had a lesser impact on all aspects of the nuclear-only DNA analyses. This problem may be exacerbated when mitochondrial and nuclear DNA analyses are combined in a total evidence approach.

Inside-out flowers of Lacandonia brasiliana (Triuridaceae) provide new insights into fundamental aspects of floral patterning.

Background and Aims. A recently described Brazilian species, Lacandonia brasiliana, shares with its longer established putative sister species from Mexico, L. schismatica, inverted floral patterning (carpels surrounding stamens) that is almost unique among angiosperms. We present a detailed ontogenetic study of L. brasiliana for comparison with other members of the tribe Triurideae (Triuridaceae) to explore the possible evolutionary origins of “inside-out” flowers.Methods. Wild-source populations of L. brasiliana were compared morphologically and ontogenetically with related species of Triurideae, using light and scanning electron microscopy.Key Results. Relatively few morphological differences separate flowers of L. brasiliana and L. schismatica. Both species have tepals with late-developing subapical appendages. In both species, the three central (almost sessile) anthers develop precociously with respect to the carpels; the anthers remain closed, and fertilization is achieved via pollen-tube growth from germinating pollen grains of the same cleistogamous flower. Carpels are initiated on fascicles.Conclusions. The close similarity between the two Lacandonia species makes it unlikely that they arose independently from two separate homeotic transformation events; they could either represent sister species or two populations of a single disjunct species. Our study underlines the problematic generic and species boundaries within Triurideae. We present an evolutionary scenario of character evolution in Triuridaceae. The inside-out Lacandonia flower could have resulted from a stabilized homeotic transformation; this hypothesis is not in conflict with constrasting theories of the origin of the Triuridaceae flower, which coincided with a shift to unisexuality. The unisexual yet highly plastic flowers that are typical of Triuridaceae could have pre-adapted the origin of the extraordinary Lacandonia morphology.

Speciation processes in putative island endemic sister bat species: false impressions from mitochondrial DNA and microsatellite data.

Cases of geographically restricted co-occurring sister taxa are rare and may point to potential divergence with gene flow. The two bat species Murina gracilis and Murina recondita are both endemic to Taiwan and are putative sister species. To test for nonallopatric divergence and gene flow in these taxa, we generated sequences using Sanger and next-generation sequencing, and combined these with microsatellite data for coalescent-based analyses. MtDNA phylogenies supported the reciprocally monophyletic sister relationship between M. gracilis and M. recondita; however, clustering of microsatellite genotypes revealed several cases of species admixture suggesting possible introgression. Sequencing of microsatellite flanking regions revealed that admixture signatures stemmed from microsatellite allele homoplasy rather than recent introgressive hybridization, and also uncovered an unexpected sister relationship between M. recondita and the continental species Murina eleryi, to the exclusion of M. gracilis. To dissect the basis of these conflicts between ncDNA and mtDNA, we analysed sequences from 10 anonymous ncDNA loci with *beast and isolation-with-migration and found two distinct clades of M. eleryi, one of which was sister to M. recondita. We conclude that Taiwan was colonized by the ancestor of M. gracilis first, followed by the ancestor of M. recondita after a period of allopatric divergence. After colonization, the mitochondrial genome of M. recondita was replaced by that of the resident M. gracilis. This study illustrates how apparent signatures of sympatric divergence can arise from complex histories of allopatric divergence, colonization and hybridization, thus highlighting the need for rigorous analyses to distinguish between such scenarios.

The Taxonomic Status of Mazama bricenii and the Significance of the Táchira Depression for Mammalian Endemism in the Cordillera de Mérida, Venezuela.

We studied the taxonomy and biogeography of Mazama bricenii, a brocket deer classified as Vulnerable by the IUCN, drawing on qualitative and quantitative morphology and sequences of the mitochondrial cytochrome-b gene. We used Ecological Niche Modeling (ENM) to evaluate the hypothesis that M. bricenii of the Venezuelan Cordillera de Mérida (CM) might have become isolated from populations of its putative sister species, Mazama rufina, in the Colombian Cordillera Oriental (CO). This hypothesis assumes that warm, dry climatic conditions in the Táchira Depression were unsuitable for the species. Our analyses did not reveal morphological differences between specimens geographically attributable to M. bricenii and M. rufina, and phylogenetic analyses of molecular data recovered M. bricenii nested within the diversity of M. rufina. These results indicate that M. bricenii should be regarded as a junior synonym of M. rufina. ENM analyses revealed the existence of suitable climatic conditions for M. rufina in the Táchira Depression during the last glacial maximum and even at present, suggesting that gene flow between populations in the CO and CM may have occurred until at least the beginning of the current interglacial period and may continue today. Because this pattern might characterize other mammals currently considered endemic to the CM, we examined which of these species match two criteria that we propose herein to estimate if they can be regarded as endemic to the CM with confidence: (1) that morphological or molecular evidence exists indicating that the putative endemic taxon is distinctive from congeneric populations in the CO; and (2) that the putative endemic taxon is restricted to either cloud forest or páramo, or both. Only Aepeomys reigi, Cryptotis meridensis, and Nasuella meridensis matched both criteria; hence, additional research is necessary to assess the true taxonomic status and distribution of the remaining species thought to be CM endemics.

A new fossil megamouth shark (Lamniformes, Megachasmidae) from the Oligocene-Miocene of the western United States

ABSTRACT The extant megamouth shark, Megachasma pelagios (Lamniformes: Megachasmidae), is a large filter-feeding fish. We here describe a new species of Megachasma, M. applegatei, sp. nov., a putative sister species of the extant M. pelagios, based on isolated teeth from late Oligocene–early Miocene (late Chattian–Aquitanian) marine deposits in California and Oregon, U.S.A. Although showing a megachasmid tooth design, teeth of M. applegatei, sp. nov., exhibit a wide morphological range and are reminiscent to those of odontaspidid sharks with strong heterodonty. Megachasma applegatei, sp. nov., could have commonly measured approximately 6 m in total length and likely had a wide range of diet, possibly including small fishes and planktonic invertebrates. The fossil record indicates that either M. applegatei, sp. nov., was broadly adapted to a wide bathymetric tolerance or was a nektopelagic feeder over both deep and shallow water habitats. SUPPLEMENTAL DATA—Supplemental materials are available for this article for free at http://www.tandfonline.com/UJVP.

Bursaphelenchus penai n. sp. (Tylenchomorpha: Aphelenchoididae), a phoretic associate of ambrosia beetles (Coleoptera: Scolytinae) from avocado in Florida

During an experimental host-plant survey for the invasive redbay ambrosia beetle, Xyleborus glabratus, in Homestead, Florida, it and three native species of ambrosia beetle, Xyleborus affinis, X. volvulus and Xyleborinus gracilis were found to be associated with a Bursaphelenchus species. This nematode species, isolated from ambrosia beetles from the tribe Xyleborini, was cultured on Monilinia fructicola or Botryotinia fuckeliana for further study and was determined to be new to science and a putative sister species to B. kiyoharai because of two apomorphic characters in males, viz., possession of a tail spike vs the typical bursal flap, and the apparent absence of the P1 ventral single papilla, both typically plesiomorphic characters for the genus. Additionally, B. kiyoharai is associated with X. serriatus suggesting that the host and microbiome associations that are shared between these two species, both of which are carried by ambrosia beetles, may have ecological and biological significance in their evolution and lineage radiation. Molecular phylogenetic analyses of the near-full-length small subunit (SSU: 18S) and the D2/D3 expansion segments of the large subunit (LSU: 28S) confirmed that B. penai n. sp. is very closely related to B. kiyoharai which is a member of the B. fungivorus clade that includes B. thailandae and B. willibaldi. Bursaphelenchus penai n. sp. and B. kiyoharai both share very similar overall spicule morphology with the B. fungivorus clade. The new species is described and can be typologically differentiated from B. kiyoharai by the position of the hemizonid and excretory pore and its different geographical and host associations.

Morphological variation and affinities of the poorly known snakeAtractus caxiuana (Serpentes: Dipsadidae).

Atractus caxiuana was recently described based on three specimens (two males and one female) from the Floresta Nacional de Caxiuanã, municipality of Melgaço, state of Pará, in the eastern portion of the Brazilian Amazon. Apart from the type series, no additional samples are known for the species. In this study, we report new specimens of A. caxiuana, providing new morphological data (meristic, morphometric, pholidosis, colour pattern, and hemipenis) and localities. We relate the variability displayed by the characters analyzed to sexual dimorphism, geographic variation, and ontogeny. Additionally, we provide detailed comparisons with A. collaris and putative sister species, and propose a new species group to accommodate this distinct and possible monophyletic assemblage.

Development and characterisation of microsatellite loci for Murina gracilis (Vespertilionidae) and cross-species amplification in two other congeneric species

Tube-nosed bats (Murina) are restricted to the forests of Asia, Papua New Guinea and northeast Australia, where they are considered highly vulnerable to habitat loss and fragmentation. Several members of the genus have been recently discovered and remain poorly known, including the putative sister species Murina gracilis and M. recondita, both of which are endemic to Taiwan. We developed novel microsatellite markers for M. gracilis and screened these in both taxa and a third Taiwanese endemic M. puta. The final number of useable markers that did not deviate from Hardy–Weinberg or linkage equilibrium was 18 for M. gracilis and M. recondita, including one that was X-linked, and nine for M. puta. We are using these markers to characterise population genetic structure and gene flow within all three taxa to inform conservation management.