Detection of <Emphasis Type="Italic">Anaplasmataceae</Emphasis> agents and co-infection with other tick-borne protozoa in dogs and <Emphasis Type="Italic">Rhipicephalus sanguineus</Emphasis> sensu lato ticks

Anaplasmosis and ehrlichiosis are of serious health concern worldwide for animals and humans. In the present study, we report the occurrence of Anaplasma platys and Ehrlichia canis in dogs and Rhipicephalus sanguineus sensu lato (s.l.) ticks from Peninsular Malaysia using a nested polymerase chain reaction assay based on amplification of the 16S rRNA gene. Anaplasma platys was detected from dogs and ticks with prevalence rates of 3.3% (8/240) and 2.9% (4/140), respectively. On the other hand, 12.9% (31/240) of the dogs and 0.7% (1/140) of the ticks were tested positive for E. canis. Additionally, co-infections of A. platys and E. canis with Babesia or Hepatozoon protozoa were also noted in this study. Double infection (E. canis?+?B. gibsoni) was observed in tick, whereas triple infections (E. canis?+?A. platys?+?B. vogeli and E. canis?+?A. platys?+?H. canis) were found in dogs. This study represents the first evidence of A. platys DNA in R. sanguineus s.l. in Malaysia.     

A preliminary molecular phylogeny shows Japanese and Austrian populations of the red mite <Emphasis Type="Italic">Balaustium murorum</Emphasis> (Acari: Trombidiformes: Erythraeidae) to be closely related

The red mite Balaustium murorum (Hermann) inhabits the Western Palaearctic realm and is well adapted to man-made structures. In Japan, B. murorum had been reported more frequently after the 1980s. A molecular phylogeny based on the nuclear 18S rRNA and mitochondrial COI genes, and including B. murorum individuals from Japan and Austria and representatives of related species from Japan showed four Balaustium species-level lineages in Japan (B. murorum, Balaustium sp. 1, Balaustium sp. 2, Balaustium sp. 3). The B. murorum lineage shared identical 18S sequence and COI haplotype with the Austrian population. Balaustium sp. 1 was detected from the Tokyo and Misaki area (Honshu Island) and was the sister group to B. murorum; the other two lineages inhabited coastal environments of Erimo, Hokkaido Island (Balaustium sp. 2) and Ainan, Shikoku Island (Balaustium sp. 3). The high genetic distances among these four lineages indicate that each lineage is a distinct species, with three of the lineages representing undescribed species. Our results are compatible with the conclusion that B. murorum was introduced to Japan from Europe, although our study did not resolve the polarity or timing of migration events.     

Distribution of Spotted Fever Group Rickettsiae in Hard Ticks (Ixodida: Ixodidae) from Panamanian Urban and Rural Environments (2007–2013)

Tick-borne rickettsiosis is an important emerging disease in Panama; to date, there have been 12 confirmed cases, including eight fatalities. To evaluate the distribution of rickettsiae in Panamanian ticks, we collected questing and on-host ticks in urban and rural towns in elevations varying between 0 and 2300 m. A total of 63 sites (13 urban and 50 rural towns) were used to develop models of spatial distributions. We found the following tick species: Rhipicephalus sanguineus s.l. (present in 54 of 63 towns and cities), Amblyomma mixtum (45/63), Dermacentor nitens (40/63), A. ovale (37/63), Rhipicephalus microplus (33/63), A. oblongoguttatum (33/63), Ixodes affinis (3/63), and Ixodes boliviensis (2/63). Rhipicephalus sanguineus s.l. was present in urban and rural towns, and other species were present only in rural towns. DNA was extracted from 408 R. sanguineus s.l., 387 A. mixtum, 103 A. ovale, and 11 A. oblongoguttatum and later tested for rickettsiae genes using PCR. Rickettsia DNA was detected in ticks from 21 of 63 localities. Rickettsia rickettsii was detected in five A. mixtum (1.29%), and Candidatus “Rickettsia amblyommii” was found in 138 A. mixtum (35%), 14 R. sanguineus (3.4%), and one A. ovale (0.9%). These results suggest that much of rural Panama is suitable for the expansion of tick populations and could favor the appearance of new tick-borne rickettsiosis outbreaks.     

Extensive cryptic diversity in the cosmopolitan sludge worm <Emphasis Type="Italic">Limnodrilus hoffmeisteri</Emphasis> (Clitellata,Naididae)

Limnodrilus hoffmeisteri Claparède, 1862 is a common freshwater worm, often regarded as an indicator of organic pollution. The taxonomic status of this species is controversial due to great variation in morphological features. Numerous morphological forms of L. hoffmeisteri are recorded in the literature, especially from Europe and North America. Today, DNA-based species delimitation assumes that species boundaries can be more objectively and effectively estimated using genetic data rather than with morphological data alone. To investigate if L. hoffmeisteri is a single species, 295 worms identified as either L. hoffmeisteri or other similar (congeneric) morphospecies, using currently accepted morphological criteria, were collected from 82 locations in the northern hemisphere. The number of primary species hypotheses (PSHs) was first explored with cytochrome oxidase subunit I (COI), the proposed DNA barcode for animal species, and with data for all specimens. Both automatic barcoding gap discovery (ABGD) and the Bayesian general mixed Yule coalescent (bGMYC) model revealed the existence of ≥25 distinct PSHs (COI lineages) in our dataset. Then, smaller samples of individuals representing major COI lineages were used for exploration of a nuclear locus, the internal transcribed spacer (ITS) region. In the ITS gene tree (81 sequences), generated by BEAST, 16 well-supported terminal groups were found, but not all of these groups were congruent with the PSHs found in the COI tree. As results across these different analyses were inconsistent, we resorted to analyzing reciprocal monophyly between gene trees and used a minimum consensus of all evidence, suggesting that there are 13 separately evolving lineages (=13 species) within our sample. The smallest uncorrected COI p-distance between these species is 12.1%, and the largest intraspecific p-distance is 16.4%, illustrating the problem of species delimitation with a DNA-barcoding gap as the sole criterion. Ten of these species are morphologically identified as “L. hoffmeisteri,” the remaining three can be attributed to morphologically distinct congeneric species. An individual from the type locality in Switzerland was designated as a neotype of L. hoffmeisteri sensu stricto. This worm belongs to one of the ten species, and this lineage is widely distributed in Europe, Asia, and North America. The remaining nine species show a mixed distribution pattern; some appear to be endemic to a restricted area, others are Holarctic. Our results provide clues to the future revalidation of some of the nominal species today placed in synonymy with L. hoffmeisteri. A BEAST analysis, based on previously published and newly generated 16S data, suggested that this complex contains also other species than those studied by us. By integrating additional genetic data, it will be possible to identify these and additional specimens in future studies of Limnodrilus, and the neotype provides a baseline for further revisions of the taxonomy of the L. hoffmeisteri complex.     

Microsatellite analysis supports the existence of three cryptic species within the bumble bee <Emphasis Type="Italic">Bombus lucorum sensu lato</Emphasis>

Mitochondrial cytochrome oxidase I (COI) partial sequences are widely used in taxonomy for species identification. Increasingly, these sequence identities are combined with modelling approaches to delineate species. Yet the validity of species delineation based on such DNA ‘barcodes’ is rarely tested and may be called into question by phenomena such as ancestral polymorphisms in DNA sequences, phylogeographic divergence, mitochondrial introgression and hybridization, or distortion of mitochondrial inheritance through such factors as Wolbachia infection. The common and widespread European bumble bee Bombus lucorum s. lato contains three distinct mitochondrial DNA lineages that are assumed to represent three cryptic species, namely Bombus cryptarum, B. lucorum s. str. and B. magnus. To test whether nuclear gene pools of the three putative species were differentiated, we genotyped 304 sympatric members of the lucorum complex (54 B. cryptarum females, 168 B. lucorum s. str. females and 82 B. magnus females, as defined using mtDNA COI haplotypes) from 11 localities spread across the island of Ireland at seven nuclear microsatellite loci. Multilocus genotypes clustered into three discrete groups that largely corresponded to the three mtDNA lineages: B. cryptarum, B. lucorum s. str. and B. magnus. The good fit of mitochondrial haplotype to nuclear (microsatellite) genotypic data supports the view that these three bumble bee taxa are reproductively isolated species, as well as providing a vindication of species identity using so-called DNA barcodes.     

Disentangling the effects of isolation-by-distance and isolation-by-environment on genetic differentiation among <Emphasis Type="Italic">Rhododendron</Emphasis> lineages in the subgenus <Emphasis Type="Italic">Tsutsusi</Emphasis>

Ecological speciation has long been noted as a central topic in the field of evolutionary biology, and investigation into the relative importance of ecological and geographical factors is becoming increasingly emphasized. We surveyed genetic variation of 277 samples from 25 populations of nine Rhododendron species within Tsutsusi subgenus in Taiwan using simple sequence repeats of expressed sequence tags. Bayesian clustering revealed four genetic lineages: (1) the Rhododendron simsii, Rhododendron kanehirai, and Rhododendron nakaharae lineage (lineage 1); (2) the Rhododendron longiperulatum, Rhododendron breviperulatum, and Rhododendron noriakianum lineage (lineage 2); (3) the Rhododendron rubropilosum lineage (lineage 3); and (4) the Rhododendron oldhamii lineage (lineage 4). Asymmetric introgressions were found from lineage 3 into lineages 1 and 2 (introgressed lineages). Genetic admixture of non-R. oldhamii species was also revealed by a neighbor-joining tree. Variation partitioning showed that environment explained much larger portions of genetic variation than geography between non-introgressed lineages (i.e., between R. oldhamii and other lineages). However, the Mantel and partial Mantel tests and the multiple matrix regression with randomization found that isolation-by-distance played a more important role than isolation-by-environment (IBE) in contributing to genetic variation in most between lineage comparisons. Nevertheless, strong IBE was found when compared between non-introgressed lineages of R. oldhamii and R. rubropilosum, suggesting post-speciation ecological divergence. Several environmental variables, including annual mean temperature, aspect, isothermality, seasonal precipitation, slope, and soil pH, could be important ecological drivers involved in reproductive isolation between R. oldhamii and non-R. oldhamii species within the Tsutsusi subgenus.     

Multi-approach analysis of the diversity in <Emphasis Type="Italic">Colletotrichum cliviae</Emphasis> sensu lato

Colletotrichum cliviae is a fungal species reported both as pathogen and endophyte with broad geographical distribution. Some purported isolates of this species have been assigned to different taxa, including Colletotrichum aracearum, Colletotrichum orchidearum and Colletotrichum. sichuanensis, for which a preliminary analysis of extensive multilocus (ACT, GAPDH, ITS, TUB2) data in this study revealed high sequence similarity with C. cliviae. We further reassessed the species delineation by using the coalescent method of the generalized mixed Yule-coalescent (GMYC) and Poisson Tree Processes (PTP). Single and multilocus gene trees strongly supported a C. cliviae s. lat. clade including the four species. This clade unfolded eight subclades grouped into three distinct lineages, but no monophyly of any of the four species. GMYC and PTP analyses confidently supported the evolutionary independence of these lineages. C. sichuanensis and C. cliviae, except one isolate, formed the largest lineage. The second lineage was made up of isolates named C. aracearum and some of C. orchidearum sharing the haplotype and the third lineage accommodated two isolates named C. cliviae and C. orchidearum. This finding suggests the synonymization of C. sichuanensis with C. cliviae whereas the taxonomic status of C. aracearum and C. orchidearum still needs clarification. This study lays great stress upon the use of comprehensive data for sequence-based characterisation of species in the C. cliviae s. lat. It also presents the first report of C. cliviae in tropical Africa and on citrus host.     

Phylogenetic analyses of <Emphasis Type="Italic">Phragmites</Emphasis> spp. in southwest China identified two lineages and their hybrids

The species/lineage delimitation and possible hybridization/introgression are prerequisites in the management of invasive organism. Phragmites australis invaded diverse habitats and displaced the native lineages in North America as a consequence of the introduction from the Eurasia. Such species threatened the biodiversity safety of the invaded regions, in particular the biodiversity hot spots. Southwest (SW) China is a biodiversity hot spot with the occurrence of Phragmites species, both native and introduced. However, the genetic identity of Phragmites species in this biodiversity hot spot remains unclear, hampering effective ecological managements. In this study, we explored the phylogenetic lineages of Phragmites species in SW China. A total of 44 accessions sampled across SW China were analyzed using two chloroplast DNA (cpDNA) markers and amplified fragment length polymorphisms. Two genetic lineages were recovered, i.e., (1) the tropical lineage which primarily consisted of native Phragmites species represented by cpDNA haplotypes I, Q, and U in relatively low altitude and (2) the common lineage including native species at higher elevations in the Hengduan Mountains as well as artificially planted species represented by cpDNA haplotype P. The between-lineage hybridization was suggested for five analyzed accessions collected from either natural or artificial habitats. The putative hybrids might have originated from the maternal native tropical lineages and paternal introduced common lineage. Our results suggest the likelihood of introgressive hybridizations in SW China and thus provided implications for future research and ecological management.     

Playing hide-and-seek with the tiny dragonfly: DNA barcoding discriminates multiple lineages of <Emphasis Type="Italic">Nannophya pygmaea</Emphasis> in Asia

We examined the utility of DNA barcode data for assessing genetic diversity of the tiny dragonfly Nannophya pygmaea Rambur in Asia. Data analyses inferred from the barcode region of cytochrome oxidase subunit I (COI) were performed with Malaysian N. pygmaea, along with the existing COI haplotypes distributed in Asia. We applied four species delimitation analyses [automatic barcode gap discovery (ABGD), generalised mixed yule coalescent (GMYC), poisson tree processes maximum likelihood (PTP_ML) and poisson tree processes simple heuristic solutions (PTP_sh)] to investigate potential lineages in this geographically widespread species. Based on our dataset, we provisionally recognize four distinct lineages or operational taxonomic units of N. pygmaea, which were represented by the taxa from Japan/Korea, China/Laos/Taiwan, Malaysia and Vietnam, respectively. Phylogenetic analyses showed two well-supported assemblages of N. pygmaea: one restricted to the taxa from Malaysia and Vietnam; and the other covering all populations further north (i.e., China, Japan, Korea, Laos and Taiwan). An extraordinarily high degree of genetic distance (up to >12 %) was detected between these two assemblages—suggesting they represent two separate species.     

Genetic diversity of <Emphasis Type="Italic">Haemaphysalis qinghaiensis</Emphasis> (Acari: Ixodidae) in western China

Haemaphysalis qinghaiensis as an endemic species in China mainly infests domestic animals and causes great harm to animals and humans in Northwestern plateau. However, there is no information about genetic diversity within the recently established populations of this tick species. Therefore, the present study analyzed the fragments of mitochondrial 16S rDNA, COI and the nuclear gene ITS1 of 56 H. qinghaiensis ticks across four regions of China which are main endemic areas of this species. Analysis showed 98.1–100% (16S rDNA), 97.9–100% (COI), 99.7–100% (ITS1) identity within individuals. For these sequences, 9, 15 and 8 haplotypes were found for 16S rDNA, COI and ITS1, respectively. Ticks from Yongjing were the most variable group, followed by Lintan, Huangyuan, and Tianzhu. Based on parallel analysis of the mitochondrial and nuclear genetic diversity of H. qinghaiensis, our results indicated that mitochondrial markers (especially COI) were much more useful than nuclear ITS for intraspecific genetic variability analysis.     

Comparative morphological analysis of two parallel mycoheterotrophic transitions reveals divergent and convergent traits in the genus <Emphasis Type="Italic">Pyrola</Emphasis> (Pyroleae,Ericaceae)

The genus Pyrola includes species with different degree of mycoheterotrophy; some species possess individuals that rely on all carbon through their associations with fungi (full mycoheterotrophy, FM), whereas some species obtain carbon through both fungi and photosynthesis by itself (partial mycoheterotrophy, PM). To investigate how plant functional traits of photosynthesis and reproduction are related to the degree of mycoheterotrophy in the initial stage of the transition from PM to FM, we determined morphological traits in FM (or nearly FM) and PM species in two independent lineages, P. picta and P. japonica complexes. We used herbarium specimens and examined leaf number, leaf area, flower number, and scape length in FM or nearly FM species (P. aphylla and P. subaphylla) and PM species (P. picta s.l. and P. japonica). We found a leaf area reduction in FM (or nearly FM) species in both lineages, suggesting that this is a convergent trait. The number of flowers was not significantly different between FM (or nearly FM) and PM species in both lineages. On the other hand, differences in the variation between FM (or nearly FM) and PM species were found in some traits between the two lineages. The FM (or nearly FM) species in one lineage only possessed rudimentary leaves, whereas that in the other linage possessed a few small, ordinary leaves in addition to those with only rudimentary leaves. The scape length of the FM (or nearly FM) species was significantly longer than that of PM species in one lineage, whereas it was shorter in the other lineage. The different and common variations are divergent and convergent traits, respectively, that could be associated with the transition to FM in Pylora. In addition, shoots of both PM species occasionally lacked ordinary leaves, possibly indicating possession of these shoots is preadaptation for the transition to FM in Pyrola.     

Identification of Japanese <Emphasis Type="Italic">Lymantria</Emphasis> species (Lepidoptera: Lymantriidae) based on DNA sequences

We examined the utility of the cytochrome c oxidase subunit I (COI) and II (COII) genes of mitochondrial DNA (mtDNA) as a tool to identify nine Japanese Lymantria species, including four Asian gypsy moth species [Lymantria dispar japonica (Motschulsky), Lymantria umbrosa (Butler), Lymantria albescens Hori and Umeno, and Lymantria postalba Inoue]. In phylogenetic trees constructed for the COI and COII genes using the maximum likelihood methods, we could identify seven out of the nine Lymantria species (L. albescens and L. postalba could not be identified). These results suggest that the DNA sequences of the COI and COII genes are useful for identifying Japanese Lymantria species.     

Comparative genome-wide analysis of repetitive DNA in the genus <Emphasis Type="Italic">Populus</Emphasis> L.

Genome skimming was performed, using Illumina sequence reads, in order to obtain a detailed comparative picture of the repetitive component of the genome of Populus species. Read sets of seven Populus and two Salix species (as outgroups) were subjected to clustering using RepeatExplorer (Novák et al. BMC Bioinformatics 11:378 2010). The repetitive portion of the genome ranged from 33.8 in Populus nigra to 46.5% in Populus tremuloides. The large majority of repetitive sequences were long terminal repeat-retrotransposons. Gypsy elements were over-represented compared to Copia ones, with a mean ratio Gypsy to Copia of 6.7:1. Satellite DNAs showed a mean genome proportion of 2.2%. DNA transposons and ribosomal DNA showed genome proportions of 1.8 and 1.9%, respectively. The other repeat types accounted for less of 1% each. Long terminal repeat-retrotransposons were further characterized, identifying the lineage to which they belong and studying the proliferation times of each lineage in the different species. The most abundant lineage was Athila, which showed large differences among species. Concerning Copia lineages, similar transpositional profiles were observed among all the analysed species; by contrast, differences in transpositional peaks of Gypsy lineages were found. The genome proportions of repeats were compared in the seven species, and a phylogenetic tree was built, showing species separation according to the botanical section to which the species belongs, although significant differences could be found within sections, possibly related to the different geographical origin of the species. Overall, the data indicate that the repetitive component of the genome in the poplar genus is still rapidly evolving.     

Genetic differentiation of <Emphasis Type="Italic">Ganaspis brasiliensis</Emphasis> (Hymenoptera: Figitidae) from East and Southeast Asia

Ganaspis brasiliensis (Ihering) (Hymenoptera: Figitidae: Eucoilinae) is a Drosophila parasitoid that has often been misidentified as G. xanthopoda (Ashmead) in recent studies. This study aims to clarify genetic differentiation of G. brasiliensis based on the nucleotide sequences of the mitochondrial cytochrome oxidase subunit 1 (CO1) gene and three nuclear DNA regions, the inter-transcribed spacers 1 and 2 (ITS1 and ITS2) and putative 60S ribosomal protein L37 (RpL37), as well as crossing experiments. Four lineages are recognized in individuals assigned as G. basiliensis by morphology, (1) individuals occurring in Japan and probably South Korea, (2) individuals from a small subtropical island of Japan, Iriomote-jima, (3) individuals from temperate lowlands of Japan and high altitude areas of Southeast Asia, and (4) individuals occurring widely in Asia, America, Hawaii and Africa. The first lineage is a specialist of Drosophila suzukii (Matsumura), a pest of fresh fruit, and also the fourth lineage has a capacity to parasitize this pest species. The first, third and fourth lineages occur sympatrically at least in Tokyo. The third and fourth lineages differed in mate choice and host use to some extent, but post-mating isolation between them was almost absent.     

Diversity of root-endophytic <Emphasis Type="Italic">Trichoderma</Emphasis> from Malaysian Borneo

Trichoderma species form endophytic associations with plant roots and may provide a range of benefits to their hosts. However, few studies have systematically examined the diversity of Trichoderma species associated with plant roots in tropical regions. During the evaluation of Trichoderma isolates for use as biocontrol agents, root samples were collected from more than 58 genera in 35 plant families from a range of habitats in Malaysian Borneo. Trichoderma species were isolated from surface-sterilised roots and identified following analysis of partial translation elongation factor-1α (tef1) sequences. Species present included Trichoderma afroharzianum, Trichoderma asperelloides, Trichoderma asperellum, Trichoderma guizhouense, Trichoderma reesei, Trichoderma strigosum and Trichoderma virens. Trichoderma asperellum/T. asperelloides, Trichoderma harzianum s.l. and T. virens were the most frequently isolated taxa. tef1 sequence data supported the recognition of undescribed species related to the T. harzianum complex. The results suggest that tropical plants may be a useful source of novel root-associated Trichoderma for biotechnological applications.     

Cryptic genetic divergence in <Emphasis Type="Italic">Scolopsis taenioptera</Emphasis> (Perciformes: Nemipteridae) in the western Pacific Ocean

We studied the phylogeny, population structure, and demographic history of Scolopsis taenioptera in the western Pacific Ocean. Using the 80 samples collected from four locations, we obtained the nucleotide sequences of mitochondrial cytochrome c oxidase subunit I and cytochrome b genes. We identified two distinct lineages showing a clear phylogeographic break that was possibly due to the Pleistocene sea-level change. One lineage was distributed in Iloilo (Philippines) and the other in Terengganu (Malaysia), Rayong (Thailand), and Ha Long Bay (Vietnam). The Terengganu and Rayong populations showed clear signs of demographic expansion; the Iloilo and Ha Long Bay populations were relatively stable or spatially expanded as geographically subdivided populations.     

Conspecific tolerance and heterospecific competition as mechanisms for overcoming resistance to invasion by an intertidal crab

The success of the invasive Asian shore crab, Hemigrapsus sanguineus, stems partly from its ability to exclude established crab species from preferred rocky and cobble intertidal habitat. Here, we assessed preference and competition for habitat types (cobble vs. sand) for H. sanguineus and two competitor species; the previous invasive green crab, Carcinus maenas, and the native rock crab, Cancer irroratus, in New England. In simple laboratory experiments, we paired similarly sized heterospecifics and conspecifics from each species, and also grouped combinations of C. maenas and H. sanguineus in a series of four-individual sets in order to dissect the outcome of intra- and inter-specific competition at different densities. Individually, all three species preferred cobble substrate. With paired conspecifics, H. sanguineus individuals would cohabitate in cobble, whereas C. maenas and C. irroratus individuals each excluded conspecifics from cobble. In heterospecific pairs, H. sanguineus excluded both C. maenas and C. irroratus from cobble. C. maenas and C. irroratus, were equally likely to exclude the other species, but rarely excluded H. sanguineus. In larger assemblages, H. sanguineus preferentially grouped under cobble, whereas C. maenas were more evenly distributed among habitat types. These observations demonstrate that conspecific tolerance and heterospecific competition can be effective, complementary mechanisms for overcoming invasion resistance. Such mechanisms help explain the well-studied success of H. sanguineus following its introduction into New England coastal habitats, and the resulting exclusion of preexisting crab species.     

Assessment of the genetic distances between some species of the family Bradybaenidae (Mollusca,Pulmonata)

On the basis of inter-simple sequence repeat (ISSR) loci and the nucleotide sequences of nuclear (18S and ITS-1) and mitochondrial genes (COI and 16S), a phylogenetic analysis of the three species of terrestrial mollusks of the family Bradybaenidae (Mollusca, Pulmonata), Bradybaena fruticum Müll., Bradybaena schrencki Midd., and Bradybaena transbaicalia Shileyko, was conducted to clarify their taxonomic status. The analysis showed that Br. fruticum was far apart from the other two species (Br. schrencki and Br. transbaicalia). The genetic distance between the latter puts in doubt their status as distinct species. It is suggested that the species Br. transbaicalia can be treated as a form of Br. schrencki var. transbaicalia.     

Establishing species and species boundaries in <Emphasis Type="BoldItalic">Sabellastarte</Emphasis> Krøyer, 1856 (Annelida: Sabellidae): an integrative approach

Sabellastarte Krøyer, 1856 (Sabellidae), a morphologically homogeneous group distributed in warm and temperate coasts of the Indo-Pacific and Caribbean Sea, is characterized by the presence of a unique combination of features. To date, the genus comprises eight species, but morphological characters traditionally used in diagnostics have shown intra-specific variability, making species boundaries and distributions unclear. The present study constitutes the first attempt to test the monophyly of Sabellastarte and its relationships to other sabellid genera by combining molecular (COI and 16S) and morphological data. Results include placement of a clade containing Stylomma, Sabella, Branchiomma and Bispira as the sister group to Sabellastarte. Phylogenetic analyses and genetic divergence among specimens from several localities around the world indicate the presence of at least six lineages within Sabellastarte. In the context of a discussion of species boundaries and diagnostic features, the distribution of some of those lineages can be explained by the presence of cryptic species and potential introductions.     

Genetic population structure and low genetic diversity in the over-exploited sea cucumber <Emphasis Type="Italic">Holothuria edulis</Emphasis> Lesson, 1830 (Echinodermata: Holothuroidea) in Okinawa Island

Understanding genetic connectivity is fundamental for ecosystem-based management of marine resources. Here we investigate the metapopulation structure of the edible sea cucumber Holothuria edulis Lesson, 1830 across Okinawa Island, Japan. This species is of economic and ecological importance and is distributed from the Red Sea to Hawai‘i. We examined sequence variation in fragments of mitochondrial cytochrome oxidase subunit I (COI) and 16S ribosomal RNA (16S), and nuclear histone (H3) at six locations across Okinawa Island. We found higher haplotype diversity for mtDNA (COI: Hd = 0.69 and 16S: Hd = 0.67) and higher heterozygosity of nDNA (H3: H _E = 0.39) in populations from the west coast of Okinawa compared to individuals from populations on the east coast (COI: Hd = 0.40; 16S: Hd = 0.21; H3: H _E = 0.14). Overall population structure was significant (AMOVA results for COI: Φ _ST = 0.49, P < 0.0001; 16S: Φ _ST = 0.34, P < 0.0001; H3: Φ _ST = 0.12, P < 0.0001). One population in the east, Uruma, showed elevated pairwise Φ _ST values in comparisons with all other sites and a marked reduction of genetic diversity (COI: Hd = 0.25 and 16S: Hd = 0.24), possibly as a consequence of a shift to a more dominant asexual reproduction mode. Recent reports have indicated that coastal development in this area influences many marine organisms, and ecosystem degradation in this location could cause the observed decrease of genetic diversity and isolation of H. edulis in Uruma. Our study should provide valuable data to help with the urgently needed management of sea cucumber populations in Okinawa, and indicates particular attention needs to be paid to vulnerable locations.