Cryptic species,taxonomic inflation,or a bit of both? New species phenomenon in Sri Lanka as suggested by a phylogeny of dwarf geckos (Reptilia,Squamata, Gekkonidae,Cnemaspis)

We evaluated the status of 16 of 22 recognized Sri Lankan Cnemaspis Strauch species, and flagged overlooked diversity with two mitochondrial (cyt b & ND2) and two nuclear markers (RAG1 & PDC) totalling 2829 base pairs. A fossil-calibrated timetree and sampling of other South Asian Cnemaspis provide insights into the diversification of the genus in peninsular India and Sri Lanka. Phylogenetic analyses consistently inferred two broad clades within South Asian Cnemaspis, with Sri Lankan species in two clades, which we call the podihuna and kandiana clades. Each Sri Lankan clade as a whole is sister to Indian taxa and nested within Indian lineages. Cnemaspis modigliani Das from Indonesia is a member of the kandiana clade. This suggests a minimum of two dispersal events between India and Sri Lanka and one between Sri Lanka/India and South-east Asia. South Asian Cnemaspis date back to at least the Eocene, in Sri Lanka to the early Miocene, with late Miocene diversification in the kandiana clade. All but one of the named species we sampled is likely to be valid, and 10 divergent unnamed lineages may warrant specific recognition. A resolution of Sri Lankan Cnemaspis taxonomy will require thorough sampling and the use of both morphological and molecular data.     

A molecular phylogeny of the freshwater-fish genus Rasbora (Teleostei: Cyprinidae) in Sri Lanka reveals a remarkable diversification—And a cryptic species

The diversity of the freshwater-fish genus Rasbora (Cyprinidae) on Sri Lanka (five species) is high compared with the four species reported from the peninsula of India, from which the island's cyprinid fauna is derived. The paucity of characters by which species of Rasbora can be phenotypically distinguished renders field identification difficult, adversely affecting the estimation of populations and distributions, with consequences for conservation and management, increasing also the risk of taxonomic inflation. From a sampling of 90 sites across Sri Lanka and based on phylogenetic and haplotype analyses of sequences of cox1 and cytb mitochondrial, and rag1 and irbp nuclear markers, we review the species diversity and phylogeography of Rasbora on the island. Molecular analyses recover, in addition to the five species previously reported, a new (cryptic) species: Rasbora adisi sp. nov. Uncorrected pairwise cox1 genetic distances between species range from 2.0 to 12.3 percent. The Sri Lankan diversification derives from a common ancestor which arrived from India during a sea-level low-stand in the mid-Miocene (15.1 Ma [95% HPD: 11.5–19.8 Ma]), when the present-day island was subaerially connected to the Indian subcontinent by a broad isthmus. This gave rise to a clade comprising five species—R. adisi sp. nov.,Rasbora armitagei, Rasbora microcephalus, Rasbora naggsi and Rasbora wilpita—with a crown age of 9.9 Ma (95% HPD: 7.1–13.3 Ma) and to a clade comprising Indian and Sri Lankan populations of Rasbora dandia, which themselves are reciprocally monophyletic. Morphological analysis of 334 specimens discriminates between most species which, however, are most reliably diagnosed by chromatic characters. The four endemic species exhibit a pattern of inter-basin dispersal via headwater capture, followed by vicariance, explaining the high diversity of the genus on the island.     

Phylogenetic relationships and molecular evolution in uropeltid snakes (Serpentes: Uropeltidae): allozymes and albumin immunology

Multilocus electrophoretic methods and microcomplement fixation comparisons of serum albumin are used to assess phylogenetic relationships among species of uropeltid snakes, to infer aspects of their population biology and biogeography, and to evaluate their relationships to other primitive snakes (Henophidia). There is very good agreement between phylogenetic inferences derived from the electrophoretic data and those derived from the albumin immunological data. Protein variation detected by electrophoresis is relatively high among 17 operational taxonomic units (OTUs) examined. The mean number of alleles per locus (5.1 across all OTUs), levels of polymorphism (25% of loci), and heterozygosity (4–6%), are typical of, or greater than, values reported for other snakes. Species of uropeltids are genetically highly differentiated, as measured by genetic distances (lowest interspecific Nei's unbiased genetic distances, 0.22-0.27 among several Sri Lankan species; 2.3 between Teretrurus of India and other uropeltines). The phylogenetic tree most consistent with both the immunological and electrophoretic data shows uropeltines from Sri Lanka to be monophyletic, but the Indian species are paraphyletic with respect to those from Sri Lanka. Rhinophis travancoricus of India is inferred to be the sister taxon to the Sri Lankan radiation. As the genera are presently understood, neither Rhinophis nor Uropeltis appears to be monophyletic. A biogeographic scenario derived from the phylogenetic hypothesis suggests an early diversification of uropeltids in India, followed by a single invasion into the lowlands of Sri Lanka. Subsequent evolution on Sri Lanka resulted in occupation of montane biotopes. Cylindrophis is the sister group to uropeltines and is considered a member of the Uropeltidae. The immunological data indicate no phylogenetic association between uropeltids and other ‘anilioid’ taxa, specifically Anilius, Loxocemus or Xenopeltis, although we cannot rule out a very remote relationship. We specifically reject the hypothesis that uropeltines and scolecophidians form a clade relative to henophidians. High levels of genetic variation and a trend toward negative F_IS values for polymorphic loci in three populations suggest generally large effective population sizes and outbreeding in these species. The niche-width variation hypothesis for allozyme loci is not supported by the uropeltid data. In comparison to other vertebrates, the relationship between Nei's genetic distance and albumin immunological distance in uropeltids suggests either conservative albumin evolution or strong differentiation at electrophoretic loci.     

The phylogenetic history of the old world butterfly subtribe Mycalesina extended: the Mycalesis (Lepidoptera: Nymphalidae) of Sri Lanka

Sri Lanka, together with the Western Ghats, is one of the world's biodiversity hotspots, yet little is known about the genetic diversity of the butterflies here. Within the framework of the recently reconstructed molecular phylogeny for the subtribe Mycalesina, we determined the phylogenetic relationships and biogeographic history of the five species of Mycalesis butterflies that are found in Sri Lanka. Sequences of Elongation Factor 1-α (EF1-α), Wingless (Wgl) and the barcode region of the Cytochrome c Oxidase sub unit 1 (COI) were used to confirm the identity of species, to resolve taxonomic queries and to infer the phylogenetic history of the group.Time-calibrated analysis of genetic data suggests that the Mycalesis species that occur in Sri Lanka diverged in the Miocene (5–23 million years ago). The results of phylogenetic analyses confirmed the following; the endemicity of Mycalesis rama to Sri Lanka; the subspecies classification of M. perseus typhlus and, the close sister relationship of the Sri Lankan M. patnia patnia to M. p. junonia of India. However, whether or not M. subdita of Sri Lanka and India are genetically similar remains unknown. Lastly, genetic evidence emerged suggesting that M. mineus forms a cryptic species complex in the Oriental region, and that in Sri Lanka, there may be occasional hybridization between M. mineus and M. perseus. As a case study of island colonization and diversification by the Mycalesis species, this study further extends our understanding of the Old World butterfly subtribe of Mycalesina.     

The taxonomy and biology of Stenosiphonium Nees (Acanthaceae)

A taxonomic revision of Stenosiphonium Nees is provided. Morphological variation within the genus is documented, the relationship between Stenosiphonium and Strobilanthes Blume is discussed, and problems of species delimitation are resolved. Three species are recognized. Stenosiphonium cordifolium (Vahl) Alston is morphologically variable and is widespread throughout peninsular India and Sri Lanka. S. setosum T. Anderson and S. wightii Bremek. are restricted endemics from the southern Western Ghats, each known from very few herbarium collections. Both S. setosum and S.wightii are recognized as rare in accordance with IUCN criteria. There is evidence to suggest that all three species of Stenosiphonium are plietesial which may explain the paucity of herbarium collections of both S. setosum and S. wightii. The implications of a plietesial lie history strategy for the assessment of the conservation status of these species is considered.     

Genus-level phylogeny of snakes reveals the origins of species richness in Sri Lanka

Snake diversity in the island of Sri Lanka is extremely high, hosting at least 89 inland (i.e., non-marine) snake species, of which at least 49 are endemic. This includes the endemic genera Aspidura, Balanophis, Cercaspis, Haplocercus, and Pseudotyphlops, which are of uncertain phylogenetic affinity. We present phylogenetic evidence from nuclear and mitochondrial loci showing the relationships of 40 snake species from Sri Lanka (22 endemics) to the remaining global snake fauna. To determine the phylogenetic placement of these species, we create a molecular dataset containing 10 genes for all global snake genera, while also sampling all available species for genera with endemic species occurring in Sri Lanka. Our sampling comprises five mitochondrial genes (12S, 16S, cyt-b, ND2, and ND4) and five nuclear genes (BDNF, c-mos, NT3 RAG-1, and RAG-2), for a total of up to 9582 bp per taxon. We find that the five endemic genera represent portions of four independent colonizations of Sri Lanka, with Cercaspis nested within Colubrinae, Balanophis in Natricinae, Pseudotyphlops in Uropeltidae, and that Aspidura + Haplocercus represents a distinct, ancient lineage within Natricinae. We synonymize two endemic genera that render other genera paraphyletic (Haplocercus with Aspidura, and Cercaspis with Lycodon), and discover that further endemic radiations may be present on the island, including a new taxon from the blindsnake family Typhlopidae, suggesting a large endemic radiation. Despite its small size relative to other islands such as New Guinea, Borneo, and Madagascar, Sri Lanka has one of the most phylogenetically diverse island snake faunas in the world, and more research is needed to characterize the island’s biodiversity, with numerous undescribed species in multiple lineages.     

Genetic divergence and evolutionary relationships in six species of genera Hoplobatrachus and Euphlyctis (Amphibia: Anura) from Bangladesh and other Asian countries revealed by mitochondrial gene sequences

To elucidate the species composition, genetic divergence, evolutionary relationships, and divergence time of Hoplobatrachus and Euphlyctis frogs (subfamily Dicroglossinae, family Ranidae) in Bangladesh and other Asian countries, we analyzed the mitochondrial Cyt b, 12S, and 16S rRNA genes of 252 specimens. Our phylogenetic analyses showed 13 major clades corresponding to several cryptic species as well as to nominal species in the two genera. The results suggested monophyly of Asian Hoplobatrachus species, but the position of African Hoplobatrachus occipitalis was not clarified. Nucleotide divergence and phylogenetic data suggested the presence of allopatric cryptic species allied to Euphlyctis hexadactylus in Sundarban, Bangladesh and several parapatric cryptic species in the Western Ghats, India. The presence of at least two allopatric cryptic species among diverged Euphlyctis cyanophlyctis in Bangladesh, India, and Sri Lanka was also suggested. In some cases, our estimated divergence times matched the paleogeological events of South and Southeast Asian regions that may have led to the divergence of Hoplobatrachus and Euphlyctis taxa. Especially, land formation at Bangladesh (15–10 Ma) may have allowed the spread of these frog taxa to Southeast Asian areas, and the aridification of central India (5.1–1.6 Ma) might have affected the gene flow of widely distributed species. The present study revealed prior underestimation of the richness of the amphibian fauna in this region, indicating the possible occurrence of many cryptic species among these groups.     

Geographic origin and taxonomic status of the invasive Privet, Ligustrum robustum (Oleaceae), in the Mascarene Islands, determined by chloroplast DNA and RAPDs

Information concerning the area of origin, genetic diversity and possible acquisition of germplasm through hybridisation is fundamental to understanding the evolution, ecology and possible control measures for an introduced invasive plant species. Among the most damaging of alien plants that are invading and degrading native vegetation in the Mascarene Islands of the Indian Ocean is the Tree Privet, Ligustrum robustum. Exact information about the geographic source of introduced material of this species is lacking, in part because Ligustrum is a taxonomically difficult genus. Native material of L. robustum ssp. walkeri from Sri Lanka, L. robustum ssp. robustum from northeastern India, and the closely related L. perrottetii from southern India was compared with introduced material from La Réunion and Mauritius using chloroplast DNA RFLP markers and random amplified polymorphic DNA (RAPDs). Sri Lankan and introduced material was monomorphic for the same cpDNA haplotype that was absent from south and northeast Indian Ligustrum. Sri Lankan and introduced material was also clearly distinguished from Indian Ligustrum by RAPDs. It was concluded that material introduced and established in the Mascarene Islands is derived from the Sri Lankan subspecies L. robustum ssp. walkeri. No geographic structuring of genetic variation within Sri Lanka was detected for this taxon, so the location(s) within Sri Lanka from which introduced material is derived could not be pinpointed. RAPDs indicate that L. robustum ssp. walkeri in Sri Lanka is more similar to south Indian L. perrottetii than to northeast Indian L. robustum ssp. robustum. Moreover, RAPDs showed that introduced material in La Réunion has undergone little or no loss of genetic diversity since introduction. However, there was no evidence that it is introgressed with germplasm from two other alien Ligustrum species present on La Réunion.     

India–Madagascar vicariance explains cascade beetle biogeography

India and Madagascar drifted apart more than 80 Mya, yet few taxonomic groups currently found in these regions bear any signature of this split. When drifting in isolation, extensive volcanic activity covered almost half of India in lava flows, likely triggering widespread extinction on the island. Consequently, most of India's rich extant flora and fauna are considered to be the result of recent Cenozoic dispersal, and no lineages are conclusively a result of ancient vicariance. Many of Madagascar's lineages also stem from either Cenozoic diversification or dispersal events, with the latter being a result of the close proximity of the island with mainland Africa. In the present study, we focus on two remarkable lineages of cascade beetles in the genera Scoliopsis and Tritonus (Coleoptera, Hydrophilidae), respectively, dwelling in the mountains of south India and Sri Lanka, as well as in Madagascar. Based on a molecular phylogeny of the family Hydrophilidae dated with eight fossils, we show that these two lineages are sister taxa, and diverged when Madagascar and Greater India (India, Sri Lanka, Seychelles) separated, suggesting a pattern of Gondwanean vicariance. The results of the present study show that, despite geological upheaval, the present‐day fauna of India still retains traces of its Gondwanan past.     

Welcome back New Zealand: regional biogeography and Gondwanan origin of three endemic genera of mite harvestmen (Arachnida, Opiliones, Cyphophthalmi)

Aim Biogeographers have long been intrigued by New Zealand’s biota due to its unique combination of typical ‘continental’ and ‘island’ characteristics. The New Zealand plateau rifted from the former supercontinent Gondwana c. 80 Ma, and has been isolated from other land masses ever since. Therefore, the flora and fauna of New Zealand include lineages that are Gondwanan in origin, but also include a very large number of endemics. In this study, we analyse the evolutionary relationships of three genera of mite harvestmen (Arachnida, Opiliones, Cyphophthalmi) endemic to New Zealand, both to each other and to their temperate Gondwanan relatives found in Australia, Chile, Sri Lanka and South Africa. Location New Zealand (North Island, South Island and Stewart Island). Methods A total of 94 specimens of the family Pettalidae in the suborder Cyphophthalmi were studied, representing 31 species and subspecies belonging to three endemic genera from New Zealand (Aoraki, Neopurcellia and Rakaia) plus six other members of the family from Chile, South Africa, Sri Lanka and Australia. The phylogeny of these taxa was constructed using morphological and molecular data from five nuclear and mitochondrial genes (18S rRNA, 28S rRNA, 16S rRNA, cytochrome c oxidase subunit I and histone H3, totalling c. 5 kb), which were analysed using dynamic as well as static homology under a variety of optimality criteria. Results The results showed that each of the three New Zealand cyphophthalmid genera is monophyletic, and occupies a distinct geographical region within the archipelago, grossly corresponding to palaeogeographical regions. All three genera of New Zealand mite harvestmen fall within the family Pettalidae with a classic temperate Gondwanan distribution, but they do not render any other genera paraphyletic. Main conclusions Our study shows that New Zealand’s three genera of mite harvestmen are unequivocally related to other members of the temperate Gondwanan family Pettalidae. Monophyly of each genus contradicts the idea of recent dispersal to New Zealand. Within New Zealand, striking biogeographical patterns are apparent in this group of short‐range endemics, particularly in the South Island. These patterns are interpreted in the light of New Zealand’s turbulent geological history and present‐day patterns of forest cover.     

The Cladocera of Sri Lanka (Ceylon), with remarks on some species

The freshwater Cladocera of Sri Lanka has been revised based on the study of over 700 zooplankton samples collected from all habitat types during 1965–1980. The cladoceran fauna is represented by six families; members of the families Polyphemidae, Leptodoridae and Holopedidae are absent. The common temperate genus Daphnia is rare.Sixty-two species have been recorded from Sri Lanka. Of these, five are new records. Remarks on a few species are given with illustrations. The distribution of Cladocera in different types of habitats is discussed. The greatest species diversity was found in ponds. The Sri Lankan fauna is numerically and in species diversity typical of tropical cladoceran fauna. It resembles the southern Indian fauna very closely except for the absence in Sri Lanka of the genera Acroperus and Camptocercus.     

Phylogeny and species delimitation of the C‐genome diploid species in Oryza

Abstract The diploid Oryza species with C‐genome type possesses abundant genes useful for rice improvement and provides parental donors of many tetraploid species with the C‐genome (BBCC, CCDD). Despite extensive studies, the phylogenetic relationship among the C‐genome species and the taxonomic status of some taxa remain controversial. In this study, we reconstructed the phylogeny of three diploid species with C‐genome (Oryza officinalis, O. rhizomatis, and O. eichingeri) based on sequences of 68 nuclear single‐copy genes. We obtained a fully resolved phylogenetic tree, clearly indicating the sister relationship of O. officinalis and O. rhizomatis, with O. eichingeri being the more divergent lineage. Incongruent phylogenies of the C‐genome species found in previous studies might result from lineage sorting, introgression/hybridization and limited number of genetic markers used. We further applied a recently developed Bayesian species delimitation method to investigate the species status of the Sri Lankan and African O. eichingeri. Analyses of two datasets (68 genes with a single sample, and 10 genes with multiple samples) support the distinct species status of the Sri Lankan and African O. eichingeri. In addition, we evaluated the impact of the number of sampled individuals and loci on species delimitation. Our simulation suggests that sampling multiple individuals is critically important for species delimitation, particularly for closely related species.     

The Indian brown mongoose,yet another invader in Fiji

A thriving population of the Indian brown mongoose (Herpestes fuscus), native to southwest India and Sri Lanka, has been discovered on the island of Viti Levu in Fiji. This is the first known introduction of this species and may derive from a pair brought from an unknown source to a private zoo in the late 1970s. They co-occur on Viti Levu with the much smaller small Indian mongoose (H. auropunctatus), and they are probably nocturnally active. No research has been conducted on their activity, diet, or impact on Fiji.     

Carving out turf in a biodiversity hotspot: multiple,previously unrecognized shrew species co‐occur on Java Island,Indonesia

In theory, competition among species in a shared habitat results in niche separation. In the case of small recondite mammals such as shrews, little is known about their autecologies, leaving open questions regarding the degree to which closely related species co‐occur and how or whether ecological niches are partitioned. The extent to which species are able to coexist may depend on the degree to which they exploit different features of their habitat, which may in turn influence our ability to recognize them as species. We explored these issues in a biodiversity hotspot, by surveying shrew (genus Crocidura) diversity on the Indonesian island of Java. We sequenced portions of nine unlinked genes in 100–117 specimens of Javan shrews and incorporated homologous data from most known Crocidura species from other parts of island South‐East Asia. Current taxonomy recognizes four Crocidura species on Java, including two endemics. However, our phylogenetic, population genetic and species delimitation analyses identify five species on the island, and all are endemic to Java. While the individual ranges of these species may not overlap in their entirety, we found up to four species living syntopically and all five species co‐occurring on one mountain. Differences in species' body size, use of above ground‐level habitats by one species and habitat partitioning along ecological gradients may have facilitated species diversification and coexistence.     

Isolation and Molecular Characterization of Entamoeba nuttalli Strains Showing Novel Isoenzyme Patterns from Wild Toque Macaques in Sri Lanka

We have proposed the revival of the name Entamoeba nuttalli for a virulent ameba strain, P19‐061405, from a rhesus macaque and located it phylogenetically between E. histolytica and E. dispar. As E. nuttalli was originally described for an ameba found in a toque macaque in Sri Lanka, the prevalence and characteristics of Entamoeba species in wild toque macaques were examined. PCR analysis of 227 stool samples from six locations showed positive rates for E. nuttalli, E. dispar, and E. histolytica of 18.5%, 0.4%, and 0%, respectively. Fifteen E. nuttalli strains were cultured successfully from five locations. The 18S ribosomal RNA gene showed only three nucleotide differences in comparison with P19‐061405 strain. In isoenzyme analysis, the pattern of hexokinase in Sri Lankan strains was different from that of P19‐061405 strains and the difference was confirmed by analysis of the genes. Hepatic inoculation of one of the Sri Lankan E. nuttalli strains in hamsters resulted in amebic abscess formation and body weight loss. These results demonstrate that E. nuttalli is prevalent in wild toque macaques and that several characteristics of the strains are unique. We conclude that use of the name E. nuttalli is appropriate for the new Entamoeba species found in nonhuman primates.     

Extended maritime jurisdiction and its impact on South Asia

Abstract

Extension of zones of national jurisdiction under UNCLOS III has increased maritime contiguity among Bangladesh, India, Pakistan, and Sri Lanka. It raises possibilities of conflict on boundary delimitation, transnational stocks and pollution and also presents opportunities for joint efforts in the exploration and exploitation of marine resources.     

Assessment of genetic diversity in Calamus thwaitesii BECC. (Arecaceae) using RAPD markers

Calamus thwaitesii Becc. is a potentially useful rattan found in the Western Ghats of India and Sri Lanka. The wild stock of this rattan species is greatly diminished due to overexploitation for the furniture industry and increasingly rare. Genetic diversity was estimated in 80 samples representing eight populations from the Western Ghats and Sri Lanka using Random Amplified Polymorphic DNA (RAPD) markers. RAPDs generated a total of 120 markers with 10 decamer primers, of which 85% were found to be polymorphic. The percentage of polymorphic loci varied from 40.00 to 60.83 and genetic distance between populations ranged from 0.0332 to 0.2777. Among the analysed populations, Goa was found to be genetically superior followed by Achenkovil, Sinharaja and Talakkaveri. Majority of the genetic diversity was distributed within populations (70.79%) and only (29.21%) among populations. Genetic relationships estimated by the unweighted pair-group method with arithmetic averaging (UPGMA) cluster analysis and principal co-ordinate analysis failed to separate Indian and Sri Lankan populations geographically into two distinct groups.     

Redescription of Amblyomma integrum adults and immature stages

Amblyomma integrum Karsch, 1879 (Acari: Ixodidae) is one of four Amblyomma Koch, 1844 species with eyes found in southern India and Sri Lanka. The immature stages of this species were poorly described. Therefore, accurate identification is difficult. Here we re‐describe the male, female, nymph and larva of A. integrum and illustrate all the stages in greater detail for the first time. A set of diagnostic morphological characters is defined to distinguish this species from other sympatric species of eyed Amblyomma in any parasitic stage of development. Adults of A. integrum parasitize mostly various larger mammals whereas nymphs and larvae use mostly larger and medium mammals. Amblyomma integrum is recorded from India (Andhra Pradesh, Goa, Karnataka, Orissa and Tamil Nadu States) and throughout Sri Lanka.     

A redescription ofDiaphanosoma modigliani Richard, 1894 (Crustacea : Daphniiformes : Sididae)and an evaluation of its occurence outside Lake Toba (Sumatra)

A pelagic species of Diaphanosoma, D. modigliani is redescribed on material from the type locality (Lake Toba, Sumatra, Indonesia). Other records of this species in tropical Asia (India, Philippine Islands, Malaysia, Sri Lanka, Thailand) are either unsubstantiated or erroneous.     

Geographical variation in acoustic signals of the planthopper, Nilaparvata bakeri (Muir), in Asia: species recognition and sexual selection

Populations of the planthopper .Nilaparvata bakeri (Muir) were sampled from the Philippines (Luzon), Indonesia (Bali), India (Bihar) and Sri Lanka (Central Province) from the grass Leersia hexandra (Schwartz), and cultured in Cardiff. Low intensity vibratory signals produced by males and females during mate location and courtship were recorded and analysed. Obvious differences were found between male calling songs of insects from the Philippines and those from the other three areas. Smaller differences were detected between the Indonesian, Indian and Sri Lankan populations. Hybrids between Philippine and Indonesian, and Indonesian and Sri Lankan, insects were obtained freely in the laboratory. Male calls of F, hybrids were variable and intermediate between parentals. Mate choice experiments showed no significant assortative mating between the different populations. There was no indication of reproductive isolation and therefore of different biological species. The assumption that different male calling songs necessarily indicate the presence of different biological species is not confirmed in this example.