Cytochrome C Oxidase Subunit I Haplotypes Research Articles

A new species of Janiralata Menzies, 1951 (Isopoda: Janiridae) from Japanese bathyal waters, with a review of the associations with invertebrates among the species of Janiralata

Abstract Isopods of the genus Janiralata Menzies, 1951 (Asellota: Janiridae) are known from shallow and deep waters of the Northern Hemisphere. Most species are free-living, but some are symbiotic on the surface of other benthic invertebrates. We obtained from three beam- and four otter-trawl hauls 18 Janiralata asellote specimens from the bathyal zone off the Pacific coasts of Tohoku, northeastern Japan. Partial nucleotide sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene (505 bp) were determined for 16 of the 18 collected specimens. Non-significant genetic differences (< 0.8%) indicated that the individuals were conspecifics. Morphological analysis indicated that all the specimens belonged to a single species, Janiralata planasp. nov. based on the unique morphology of the anterolateral and distolateral projections on the head and pleotelson, surface and lateral margins of the body, antennula article 1, and of the male pleopod 1. Six of the 18 collected specimens were found attached to the surface of an unidentified sea anemone (Hormathiidae), ten to the solasterid starfish Crossaster borealis Fisher, 1906 and two were obtained from the bottom sediment. Three COI haplotypes of the new species were shared by individuals from two neighboring sites, which suggests that genetic isolation occurs on a relatively narrow horizontal scale, although the bathymetric distribution range was rather wide. We describe a case study of the geographic and bathymetric distributions of an asellote species with low dispersal ability and its genetically differentiated populations. Such approach could help elucidate the diversity and distribution of deep-sea organisms and their diversification processes.

Range extensions of Pacific bone-eating worms (Annelida, Siboglinidae, Osedax).

First described in 2004 off California, Osedax worms are now known from many of the world's oceans, ranging from 10 to over 4000 m in depth. Currently, little is known about species ranges, since most descriptions are from single localities. In this study, we used new sampling in the north-eastern Pacific and available GenBank data from off Japan and Brazil to report expanded ranges for five species: Osedaxfrankpressi, O.knutei, O.packardorum, O.roseus and O.talkovici. We also provided additional DNA sequences from previously reported localities for two species: Osedaxpriapus and O.randyi. To assess the distribution of each species, we used cytochrome c oxidase subunit I (COI) sequences to generate haplotype networks and assess connectivity amongst localities where sampling permitted. Osedaxfrankpressi, O.packardorum, O.priapus, O.roseus and O.talkovici all had one or more dominant COI haplotypes shared by individuals at multiple localities, suggesting high connectivity throughout some or all of their ranges. Low ΦST values amongst populations for O.packardorum, O.roseus and O.talkovici confirmed high levels of gene flow throughout their known ranges. High ΦST values for O.frankpressi between the eastern Pacific and the Brazilian Atlantic showed little gene flow, reflected by the haplotype network, which had distinct Pacific and Atlantic haplotype clusters. This study greatly expands the ranges and provides insights into the phylogeography for these nine species.

Theama japonica sp. nov., an Interstitial Polyclad Flatworm Showing a Wide Distribution along Japanese Coasts.

We establish a new interstitial polyclad species, Theama japonica sp. nov., based on specimens collected from coarse-sandy habitats in three Japanese main islands (Hokkaido, Honshu, and Shikoku) along the coasts of the Pacific Ocean and the Sea of Japan. Theama japonica is characterized by i) two pairs of cerebral eyespots and four to six precerebral eyespots; ii) eosinophilic secretion glands distributed in the distal half of the inner ventral part of the prostatic vesicle; iii) a conical penis papilla, bent up dorsally, with a sclerotized inner wall; iv) the prostatic sheath with an inner angular fold on the dorso-distal side; and v) the external cilia longer dorsally than ventrally. Partial sequences of the cytochrome c oxidase subunit I (COI) gene from 20 specimens collected at eight localities along Japanese coasts represented 19 haplotypes. The uncorrected p-distances among these COI haplotypes fell within intraspecific variations observed in other polyclads. A network analysis based on these COI haplotypes suggested a geographically non-cohesive genetic structure of the species, possibly indicating the species' high dispersibility. Molecular phylogenetic analyses based on a concatenated dataset of 18S and 28S rDNA sequences showed T. japonica formed a clade with other Theama species. The resulting tree also indicates that our new species is more closely related to Theama sp. from Colombia than species from Panama and Croatia.

Encountering the morphological and molecular complexity in the bramble shark Echinorhinus cf. E. brucus (Bonnaterre 1788) from the Oman Sea

Abstract This is the first simultaneous morphological and barcoding characterization with the cytochrome c oxidase subunit I (COI) of the bramble shark Echinorhinus from the coast of Oman. The morphology of the specimen was consistent with previous records of Echinorhinus from the Gulf of Oman and the Arabian Sea (Northwestern Indian Ocean). However, the new COI haplotype clustered together with homologous sequences of specimens from India. The specimen from Oman distinguished morphologically and genetically from an E. brucus from the Western Atlantic Ocean on the shape and size of the dermal denticles, the proportions of twelve morphometric measurements (differences ⩾3%) and the genetic p-distance = 3.8% of the COI fragment. The haplotype reported here increases the genetic diversity in genus Echinorhinus in the Northwest Indian Ocean, demonstrates conspecificity between specimens from Oman and Echinorhinus cf. E. brucus distributed in India and extends its range of distribution. The limited morphological and molecular data available constrained assigning our specimen to other than Echinorhinus cf. E. brucus (Bonnaterrez, 1788). Our findings highlight the urgent need of morphological review, redescription and the assignment of a neotype in order to guarantee accurate species identification and thus effective conservation measures for these deep-sea sharks. The existence of a third living species in the genus is briefly discussed.

The atlas of mitochondrial genetic diversity for Western Palaearctic butterflies

AbstractMotivationButterflies represent a model in biology and a flagship group for invertebrate conservation. We provide four new resources for the Western Palaearctic butterflies: (1) an updated checklist comprising 552 species; (2) a curated dataset of 32,126 mitochondrial cytochrome c oxidase subunit I (COI) sequences for 532 species, including a de novo reference library for the Maghreb (Morocco, northern Algeria and Tunisia) and Macaronesia (Azores, Madeira and Canary Islands); (3) seven indexes of intraspecific genetic variation (IGV): observed and expected number of haplotypes, haplotype and nucleotide diversity, two fixation indexes and maximum p‐distance; and (4) species‐level maps illustrating the distribution of COI variability and haplotype networks. The updated checklist will be fundamental for any application dealing with butterfly diversity in the Western Palaearctic. The IGV indexes provide measures for genetic polymorphism and spatial structure and represent proxies for dispersal capacity. These resources will facilitate comparative studies of macrogenetics, foster integrative taxonomy and aid conservation strategies.Main types of variables containedA complete species checklist in table format, 32,126 mitochondrial DNA barcodes provided with metadata (species membership, WGS84 coordinates and sequence length) and a book in PDF format, including the IGV atlas and indexes, are provided.Spatial location and grainThe checklist encompasses Europe up to the Urals in the east, north Macaronesia (the Azores, Madeira and the Canary Islands) and the Maghreb (Morocco, northern Algeria and Tunisia). COI sequences have been retained in the geographical interval of −31.3 to 67.5° of longitude and 27.5–71.2° of latitude.Time period and grainCOI sequences originate from studies published between 1998 and 2022 and from de novo sequencing of 2541 specimens done between 2007 and 2022.Major taxa and level of measurementButterflies (Lepidoptera: Papilionoidea), analysed from individual to species level.Software formatData and functions to manage the dataset are provided in the iodatabase R package (https://github.com/leondap/iodatabase) and in Dryad (https://doi.org/10.5061/dryad.9w0vt4bjj).

Genetic and morphological evidence indicates the persistence of Japanese mitten crab mitochondrial DNA in Europe for over 20 years and its introgression into Chinese mitten crabs

Cryptic biological invasions are largely unrecognised, leading to an underestimation of the number of invading taxa and their potential impacts. The Chinese mitten crab, Eriocheir sinensis, is a highly invasive species with serious economic and ecological impacts in Europe. Recently, mitochondrial DNA (mtDNA) of the Japanese mitten crab, E. japonica, has been discovered in populations from The Netherlands, Poland and Germany, but the taxonomic status and time of introduction of specimens carrying this mtDNA are uncertain. To this end, we investigated the morphology and variation of the mitochondrial cytochrome c oxidase subunit I (COI) gene of mitten crabs collected in central-western Europe between 1998 and 2020. Mitten crabs from Belgium harboured a Japanese mitten crab COI haplotype in 33% to 65% of individuals, even in our earliest samples from 1998. All other studied populations carried only Chinese mitten crab COI haplotypes. Morphologically, many of the juvenile Belgian mitten crabs showed intermediate traits between the two species, while all investigated adult mitten crabs, regardless of their mitochondrial haplotype or country of origin, were morphologically assigned to E. sinensis. This intermediate morphology of the juveniles and genetic-morphological discrepancy of adults suggests that Japanese mitten crabs introgressed with Chinese mitten crabs, which could have happened both before and after the introduction of mitten crabs to Europe. A specific Chinese mitten crab COI haplotype, found in Belgium, was previously only known from Vladivostok (Russia), where Chinese and Japanese mitten crab hybrids naturally occur. This Far East region is, therefore, a plausible source for at least part of the mitten crab mitochondrial diversity in Belgium.

Genetic diversity and demographic history of the Old World Bollworm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae), in Ethiopia inferred from mitochondrial gene sequences

The Old World bollworm, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae), is a globally distributed agricultural and horticultural insect pest. Despite the economic importance of this insect in Ethiopia, its genetic diversity and demographic history are poorly understood. We examined the nucleotide variation of the mitochondrial cytochrome c oxidase subunit I (COI) gene fragment of 74 H. armigera individuals from six collection sites in Ethiopia. We recorded 15 COI haplotypes in H. armigera, ten globally shared and five exclusive to Ethiopia (HaET15, HaET14, HaET10, HaET7, and HaET4). Haplotype HaET1 was the most widely geographically distributed and frequent (71.62%). Analysis of molecular variance (AMOVA) revealed a high and significant level of variation within H. armigera populations (θST = −0.0135). Negative values of the neutrality test and nonsignificant index of mismatch distribution supported the demographic expansion of H. armigera populations in Ethiopia; furthermore, this was also supported by the nonsignificant values of the sum of squared deviations (SSD) and raggedness index (r). The high genetic variation and population expansion of H. armigera have immense implications for devising locally adapted management strategies in area‐wide integrated pest management IPM programs. However, a comprehensive study of H. armigera genetic diversity and population structure using various molecular markers is needed for future confirmation.

Pliocene-Pleistocene dispersal bring along low inter species diversity between Vimba species based on multilocus analysis

This study investigates phylogenetic and phylogeographic relationships of Vimba species using mitochondrial cytochrome b (cyt b) (1023 bp) and cytochrome c oxidase subunit I (COI) barcoding region (652 bp) genes. Ninety-one samples from 36 populations for the cyt b gene and 67 samples from 20 populations for the COI were analyzed. We identified 29 haplotypes and calculated overall haplotype diversity as Hd: 0.907 ± 0.015 for cyt b. We also identified 13 COI haplotypes and calculated overall haplotype diversity as 0.826 ± 0.026 for this marker. The phylogenetic analysis of Vimba species reveals the presence of four clades, based on concatenated cyt b and COI sequences. The first and second clade consist of Vimba vimba Western lineage, and Vimba vimba Caspian lineage, while the third and fourth clade consist of Vimba mirabilis and Vimba melanops. Based on haplotype network analyses and phylogeographic inferences, the Vimba genus is monophyletic, and its species dispersed in the Pleistocene era.

Loss of mitochondrial diversity in invasive populations of Asian honey bees, Apis cerana (Hymenoptera: Apidae), in the Austral‐Pacific

AbstractIntroduced species are a significant global concern because they can disrupt native ecosystems and cause economic damage to agricultural systems and infrastructure. The mitochondrial DNA (mtDNA) diversity of invasive populations can help to identify the source and number of incursions and reveal whether these populations have maintained standing genetic variation that might allow mitochondrial genomes to adapt to new environments. We investigated mitochondrial diversity in native and invasive populations of the Asian honey bee (Apis cerana). We generated 144 sequences of mitochondrial cytochrome c oxidase subunit I (COI) from samples spanning three invasive populations (Australia, New Guinea and Solomon Islands) and three native‐range Indonesian populations (Java, Sulawesi and Flores). We then analysed these sequences together with published COI sequences to evaluate changes in mtDNA diversity associated with the recent spread of A. cerana across the Austral‐Pacific region. Mitochondrial COI diversity was consistent with invasive A. cerana populations having endured severe founder event bottlenecks during colonisation of New Guinea from Java, and then again during colonisation of Australia and the Solomon Islands from New Guinea. Mitochondrial diversity of native‐range Indonesian populations was high (26 COI haplotypes detected), particularly on Java (20 haplotypes), while all samples from invasive populations carried the same COI haplotype. Indeed, the single COI haplotype of invasive A. cerana populations is consistent with invasive populations being founded by just one, or very few, queens, as previously suggested from nuclear markers. Further investigation is needed to understand the fitness effects, if any, of mitochondrial diversity loss in invasive A. cerana populations of the Austral‐Pacific.

Population Genetic Structures and Demographic Expansion of the Exotic Jellyfish Carybdea brevipedalia in Korean Coasts Inferred from Mitochondrial COI Analysis.

Carybdea brevipedalia Kishinouye, 1891 is a poisonous jellyfish that usually occurs only in Japanese coastal regions. However, it was recently found on the Korean coast, thus expanding its known geographical range. In this study, we analyzed the population genetics and demographic histories of 113 C. brevipedalia specimens from the southern and eastern coastal regions of Korea by sequencing mitochondrial DNA cytochrome c oxidase subunit I (COI). We identified 42 C. brevipedalia COI haplotypes with high genetic diversity and a significant genetic structure. Populations were highly differentiated based on geographic location and distinctly divided into A and B clades. The results of Mantel tests indicated that geographic distance influenced the genetic distance between the two clades. Moreover, demographic analyses (neutrality tests) and the star-like profile of the Templeton, Crandall, and Sing (TCS) haplotype network indicated that C. brevipedalia had recently expanded into the southern and eastern coastal regions of Korea. These findings suggest that C. brevipedalia populations along the Korean coast have significant genetic differentiation that could be influenced by geographic isolation and subsequent adaptation to regional ecological conditions.

Brazilian invasion and dispersion routes of the cotton boll weevil

AbstractThe cotton boll weevil, Anthonomus grandis (Coleoptera: Curculionidae), is the major insect pest of cotton in the Americas. Although the demographic and dispersion patterns of the cotton boll weevil in the USA and Mexico were well characterized, in Brazil the information is scarce and does not include the main cotton‐producing regions of the country. Thus, our objective was to apply a phylogeographic approach sequencing a gene fragment of the cytochrome c oxidase subunit I (COI) to estimate the genetic diversity, identify the genetic strains, and describe the dispersion routes of the cotton boll weevil in the Brazilian territory. The COI marker revealed a low genetic diversity of the cotton boll weevil with only two haplotypes in Brazil. The haplotypes are genetically related to individuals from the eastern region of the USA and Mexico, which suggests a recent invasion event in Brazil. The genetic structure of COI haplotypes indicates a low dispersion of boll weevils between north/northeast and midwest/southeast regions of Brazil. The genetic drift and low gene flow during the insect expansion explain the COI haplotype distribution. The genetic structure of the cotton boll weevil in Brazil is novel information and provides valuable insight for the design of suitable pest management practices against this key pest species in the country.

Weak population‐genetic structure of a widely distributed nematode parasite of frogs in the western Palearctic

The genetic structure of parasite populations is affected by various factors such as host–parasite interactions, life-history strategies, and the evolutionary histories of both interacting organisms. In this study, we investigated the distribution, prevalence, and population-genetic structure of Icosiella neglecta (Spirurida, Onchocercidae), a nematode parasite found in Ranidae frogs. We reported this parasite from eight species of water frogs (genus Pelophylax) in Europe, the Middle East, and Central Asia. Its prevalence across investigated localities varied from 3.03% to 95.83%. Based on nucleotide variation in a 28S ribosomal RNA gene, all investigated I. neglecta sequences formed a well-supported phylogenetic clade and were placed in the sister position to the genus Ochoterenella. Despite the substantial genetic variability in a mitochondrial cytochrome c oxidase subunit I (COI) fragment (33 unique haplotypes recognized among 91 sequences), we found only weak population-genetic structure across the study area. There was no obvious association of COI haplotypes with geography, except haplotypes from eastern Turkey, Lebanon, and Iraq which formed a homogeneous, albeit only weakly differentiated group. The historical demographic analyses suggest that the species underwent a sudden and relatively recent population expansion. According to our results, we assume that the population-genetic structure of I. neglecta might be linked to the evolutionary history and dispersal of its dipteran vectors than water frog hosts.

Population genetics and conservation of recently discovered springsnails in Arizona

Establishing baseline geographical distributions of extant genetic diversity is increasingly important for future conservation efforts of freshwater species. We analyse the mitochondrial cytochrome c oxidase subunit I (COI) gene to taxonomically characterize 233 samples from recently discovered springsnail populations throughout 17 sites in Arizona, USA. A total of 28 unique COI haplotypes were recovered, with the number of haplotypes ranging from 1 to 4 by population in Arizona. Phylogenetic analyses resulted in haplotypes from 13 of 17 locations in Arizona being successfully identified to species, with five described and three undescribed species in the genus Pyrgulopsis (Hydrobiidae). Future work will require in-depth morphological work to clarify the taxonomic status of these putatively novel species. Importantly, among recovered species, we identified haplotype diversity of the critically endangered Three Forks springsnail, Pyrgulopsis trivialis, in the eastern Gila watershed, which will inform wildlife managers in deciding which source populations to use in reintroduction efforts. We discuss possible causes for observed population structure of Arizona's springsnail populations, with suggestions for the future sampling schemes necessary for the conservation of this uniquely important freshwater mollusc.

Genetic diversity, population structure and rapid early detection of the parasitoid Anastatus orientalis (Hymenoptera: Eupelmidae) inside eggs of spotted lanternfly (Hemiptera: Fulgoridae)

AbstractSpotted lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae), is a native pest of more than 70 plant species in China. To date, only two parasitoids are well recognised for biological control of this pest, among which, the egg parasitoid Anastatus orientalis (Hymenoptera: Eupelmidae) is known to account for approximately 69% of parasitism observed in the field. Variable emergence timings have been observed in different geographic populations in this parasitoid. To understand the genetic structure among A. orientalis populations we sequenced partial mitochondrial Cytochrome c oxidase subunit I (COI) sequences and the D2 region of the nuclear 28S gene across five regions of China (Beijing, Gansu, Shaanxi, Ningxia, Shandong). We identified 16 COI haplotypes among these populations and the lowest level of haplotype diversity was found in the Ningxia population Hd = 0.583 ± 0.133). Results from the analysis of molecular variance indicated that most of the genetic variation is present within the populations. Phylogenetic analysis demonstrated a weak geographic structure among the populations. To evaluate parasitism rapidly, we developed a 558‐bp species‐specific molecular marker which could efficiently detect parasitoids of any developmental stage inside their host eggs. Sensitivity analysis of markers revealed that our assay could detect as low a DNA template concentration as 0.025 pg/μl. This method provided more rapid and accurate detection of parasitoids compared with regular natural rearing methods.

Toward the conservation of the endemic monotypic fish genus Aulopyge from the Balkan Dinaric karst: Integrative assessment of introduced and natural population.

The complex biogeographical history of the Balkan Peninsula caused remarkable freshwater fish diversity and endemism, among which Cyprinidae fish dominate. The Dinaric karst was a Pleistocene refugium and it harbors ancient and endemic cyprinids, including Aulopyge huegelii, a sole representative of its genus. Being highly distributionally restricted, it faces various threats that promote a critical decline in population abundance and even population extinction. Phenotypic and molecular diversity of the introduced (Šator Lake, Šator Mountain) and natural (Studena River, Duvanjsko Polje) populations of Dalmatian barbelgudgeon from Bosnia and Herzegovina was studied by using two mitochondrial genes and morphometric traits (linear and geometric morphometrics). Nonparametric ANOVA showed that two analyzed populations significantly differed in six linear measurements, except snout length and postorbital head length. Contrary to centroid size, two populations were found to be significantly different in body shape. Deformation grids indicated that individuals from Studena River are characterized by wider and slightly shorter body comparing to individuals from Šator Lake. Incongruence in cytochrome c oxidase subunit I (COI) and cytochrome b (cyt b) mitochondrial DNA (mtDNA) variation was observed since a common COI haplotype was observed, while four and three cyt b haplotypes were registered in Šator Lake and Studena River, respectively. Since it was demonstrated that cyt b mtDNA was a faster evolving gene, we encourage its use in intraspecies studies, especially for evaluating the connectivity of fragmented populations and for studying the evolutionary footprint of the processes incorporated into the distinctive evolution of Aulopyge. Finally, findings herewith provide a firm basis for designing a long‐term sustainable conservation strategy for endemic species in Dinaric karst.

Genetic and Morphologic Variation in a Potential Mosquito Biocontrol Agent, Hydrochara Affinis (Coleoptera: Hydrophilidae)

Hydrochara affinis (Coleoptera: Hydrophilidae), a water scavenger beetle, was recently identified as a natural and effective agent for biological mosquito control; it was reported to exhibit high rates of mosquito larvae predation. However, maintaining the quality (i.e., natural ecological attributes, such as genetic variation) of laboratory-reared populations is essential for ensuring the long-term success of biological control programs. Accordingly, here, we aimed to use mitochondrial Cytochrome c oxidase subunit I (COI) sequences to document the genetic diversity, population structure, and phylogenetic position of natural and lab-reared H. affinis populations in South Korea and use geometric morphometric analysis to investigate the populations’ morphological divergence. The natural H. affinis populations possessed high genetic diversity and numerous COI haplotypes, suggesting that these populations were healthy and could be directly applied to mosquito habitats without alterations to their natural genetic attributes. The lab-reared populations also possessed high genetic diversity and, thus, the potential for high adaptive capacity to new environments. Although no distinct population genetic structures were observed, quantitative variation was observed in the body shape of both the natural and lab-reared populations. The high levels of genetic and morphologic variation observed in the H. affinis populations examined here indicate the species’ favorable conservation status, genetic diversity, adaptive capacity, and, thus, “suitability” for field application as an effective mosquito control agent.

Evidence for multiple maternal lineages of the invasive pest Helicoverpa armigera in Argentina

The polyphagous Helicoverpa armigera (Lepidoptera: Noctuidae) causes significant agricultural losses worldwide. This pest has recently expanded its range into South America. Since its first report in north-western Argentina, H. armigera has already been detected in geographically distant locations within the country. In this work, mitochondrial cytochrome c oxidase subunit I (COI) and cytochrome b (Cytb) sequences obtained from H. armigera individuals collected in central and northern Argentina were analyzed to explore genetic variability. Five COI haplotypes and three Cytb haplotypes were detected among ten sequenced specimens (seven haplotypes if considering concatenated sequences). Thus, even with a limited number of samples, a high number of haplotypes was found, suggesting the occurrence of several female founder lineages of H. armigera in the country. With one exception, these mtDNA haplotypes were shared with H. armigera specimens from the neighboring countries of Brazil, Paraguay and/or Uruguay. However, the impact of natural migration and intercontinental agricultural trade on the establishment and spread of H. armigera in Argentina and other Latin-American countries is still a matter of research.

Range expansion in the land snail species Carinigera buresi (Clausiliidae): long-distance dispersal by ancient marble transport?

The clausiliid Carinigera buresi has a distribution that, considering its low vagility, is enigmatically disjunct. Carinigera buresi pharsalica occurs in Thessaly in central Greece, over 200 km outside the main range of C. buresi in NE Greece – SW Bulgaria. This range disjunction has led to the hypothesis that in antiquity the ancestors of C. buresi pharsalica were transported by the marble trade from NE Greece to Thessaly. To explore this hypothesis, we included samples from the NE Greek island of Thasos (the only area within the range of C. buresi from which marble was widely transported in antiquity) in a time-calibrated molecular phylogeny of C. buresi. Maximum likelihood (GARLI) and time-calibrated Bayesian (BEAST) analyses of mitochondrial cytochrome c oxidase subunit I (COI) from samples collected throughout the range of C. buresi indicate that the ancestral C. buresi pharsalica COI lineage reached Thessaly not from Thasos, but from the NE Greek mainland. This happened between 1,740 and 83,000 yr BP according to the BEAST analyses. Although this time interval does not entirely pre-date the earliest phase of marble transport in the region, we consider this mode of dispersal unlikely for two reasons: (1) our data show that C. buresi pharsalica must be older than estimated on the basis of geological calibration points; (2) our findings indicate that the dispersal event that brought C. buresi pharsalica to Thessaly did not follow ancient marble transport routes. We suggest dispersal via floating objects or aerial dispersal by birds as alternative hypotheses to dispersal by the historic marble trade. Unexpectedly, our phylogenetic analyses also showed the COI haplotype of C. buresi from the city of Kavala to be nested among COI haplotypes from northern Thasos. This suggests possible human-aided dispersal of C. buresi from Thasos to Kavala.

Assessing the Taxonomic Validity of Austrodiplostomum spp. (Digenea: Diplostomidae) through Nuclear and Mitochondrial Data

Adults of the genus Austrodiplostomum are parasites in cormorants of the New World, whereas metacercariae are parasites from eye globe and brain of freshwater and brackish water fishes. In this study, specimens of Austrodiplostomum mordax from South America (type-species) were analyzed together with other specimens of Austrodiplostomum spp. collected from several locations across Middle America and North America. Partial DNA sequences of the mitochondrial gene cytochrome c oxidase subunit I ( COI), the internal transcribed spacers ( ITS1, ITS2, and 5.8S gene), and the D2-D3, domains of the large subunit ( LSU) of nuclear ribosomal DNA, were generated for both developmental stages and compared with available sequences of Austrodiplostomum spp. Phylogenetic analyses inferred with each molecular marker using maximum likelihood and Bayesian inference revealed the existence of 4 lineages representing 2 described species, A. mordax and Austrodiplostomum compactum (syn. Austrodiplostomum ostrowskiae) and 2 undescribed species of Austrodiplostomum recognized in previous studies. The COI haplotype network inferred with 172 sequences detected 28 haplotypes divided into 4 clusters, separating each other by 33 and 40 substitutions and with a genetic divergence ranging from 9 to 12%. The largest group included specimens identified as A. compactum plus those identified as A. ostrowskiae, supporting the synonymy of both species. As a result, we conclude that A. compactum is widely distributed across the Americas, in locations of the United States, Mexico, El Salvador, Honduras, Costa Rica, Venezuela, Peru, and Brazil. The other 2 undescribed species of the genus Austrodiplostomum were previously recorded in the United States and now are reported in Mexico. These 2 species cannot be described because adult forms have not been found in their definitive hosts. Finally, the species A. mordax has been found only in some lakes from Argentina, and it was validated in this study through molecular analyses.

DNA Barcoding Reveals Identity of Mugilids (Teleostei: Mugiliformes) from the East China Sea along the Coast of Zhejiang, China

Dramatically intensified economic development in the coastal areas of Zhejiang, China, has posed unprecedented challenges and pressures on the native coastal marine biota. Mugilids (Teleostei: Mugiliformes) are widely distributed across marine, brackish, and freshwater habitats in tropical, subtropical, and temperate regions of the world. In this study, DNA sequences of a consensus length of 611 nucleotides in mitochondrial Cytochrome c Oxidase subunit I (COI ) were used to identify 144 mugilid samples collected from various intertidal waters along the coast of Zhejiang in the East China Sea. A total of 35 distinctive COI haplotypes was scored. Results of neighbor-joining ( NJ) clustering clearly support classification of seven species, in agreement with morphological diagnosis: Planiliza affinis, Planiliza haematocheila, Planiliza sp. H, Planiliza subviridis, Crenimugil sp. A, Osteomugil cunnesius, and Mugil sp. C. Planiliza affinis was found to be the dominant mugilid in the coastal waters. This research provides critical scientific data for understanding the identity and abundance of mugilids and managing their fishery resources and habitats along the coast of Zhejiang.