DNA Barcoding of Scavenging Amphipod Communities at Active and Inactive Hydrothermal Vents in the Indian Ocean

Hydrothermal vent areas have drawn increasing interest since they were discovered in 1977. Because of chemoautotrophic bacteria, they possess high abundances of vent endemic species as well as many non-vent species around the fields. During the survey conducted by the Bundesanstalt für Geowissenschaften und Rohstoffe (Federal Institute for Geosciences and Natural Resources, BGR) to identify inactive polymetallic sulfide deposits along Central and Southeast Indian Ridges, the INDEX project studied the scavenging amphipod community at three newly discovered hydrothermal fields. A sample consisting of 463 representatives of Amphipoda (Malacostraca: Crustacea) was collected by means of baited traps in active and inactive vents of three different sites and subsequently studied by both morphological and genetic methods. Molecular methods included the analysis of two mitochondrial (cytochrome c oxidase subunit I [COI] and 16S rRNA) and one nuclear (18S rRNA) genes. By six delimitation methods, 22 molecular operational taxonomic units (MOTUs) belonging to 12 genera and 10 families were defined. The existence of potential species complexes was noted for the representatives of the genus Paralicella. The inactive site, where 19 species were found, showed higher species richness than did the active one, where only 10 taxa were recorded. Seven genera, Ambasiopsis, Cleonardo, Eurythenes, Parandania, Pseudonesimus, Tectovalopsis, and Valettiopsis, were observed only at inactive sites, whereas Haptocallisoma, was collected exclusively at active ones. The species Abyssorchomene distinctus (Birstein and Vinogradov, 1960), Hirondellea brevicaudata Chevreux, 1910, and Hirondellea guyoti Barnard and Ingram, 1990, have been previously reported from vent sites in the Atlantic or Pacific oceans. The present study provides the first report of Eurythenes magellanicus (H. Milne Edwards, 1848) and five other already described species in the Indian Ocean. The addition of 356 sequences strongly increases the number of amphipod barcodes in reference databases and provides for the first time COI barcodes for Cleonardo neuvillei Chevreux, 1908, Haptocallisoma abyssi (Oldevig, 1959), Hirondellea guyoti, Tectovalopsis fusilus Barnard and Ingram, 1990, and the genera Haptocallisoma, Pseudonesimus, and Valettiopsis.

A morphological and molecular reinvestigation of Janickina pigmentifera (Grassi, 1881) Chatton 1953 – an amoebozoan parasite of arrow-worms (Chaetognatha)

Amoebozoan parasites of arrow-worms (Chaetognatha) were isolated from their hosts living in plankton of the Bay of Villefranche (Mediterranean Sea). Based on the light microscopic characters, the amoebae were identified as Janickina pigmentifera (Grassi, 1881) by their limax locomotive form and due to the presence of the intracellular symbiont, Perkinsela amoebae, surrounded by a layer of pigment granules. Sequences of the 18S rRNA gene of both J. pigmentifera and its symbiont were obtained for the first time. The molecular phylogenetic analyses of 18S rRNA gene placed J. pigmentifera within the genus Neoparamoeba, a taxon also characterized by the presence of a symbiont, known as Perkinsela amoebae-like organism (PLO). The 18S rRNA gene sequence of P. amoebae from J. pigmentifera grouped with the sequences of 18S rRNA genes of PLOs from Neoparamoeba branchiphila and Neoparamoeba invadens. The first photo documentation of the light microscopic features of J. pigmentifera, such as locomotive form, the morphology of the nucleus and P. amoebae have been provided. The new results support the affinity of J. pigmentifera with the family Paramoebidae suggested previously based on the presence of PLO. In contrast to Janickina, typical members of Paramoebidae (Neoparamoeba and Paramoeba) have a flattened, dactylopodial locomotive form. This discrepancy in morphology can be explained by the obligate parasitic lifestyle of Janickina.

Two New Species of the Genus Longipedia Claus, 1863 (Copepoda: Harpacticoida: Longipediidae) from Korea, with an Update and a Key to Species

Benthic harpacticoids were collected from Korean waters. Two species were identified as members of the genus Longipedia Claus, 1863, because they have an extremely elongated distal segment of the P2 endopod. Longipedia koreana sp. nov. is morphologically most closely related to L. nichollsi Wells, 1980 and L. scotti Sars, 1903, but it can clearly be distinguished from both species based on the following morphological characteristics: P1 coxa with strong spinules near the outer margin and the distal element being much bigger than the proximal elements, P2 coxa with a small inner seta on the anterior surface, P4 exopod first segment without an inner element, and the P5 with a rectangular exopod (more than 3.5 times as long as wide). L. ulleungensis sp. nov. is similar to L. brevispinosa Gurney, 1927, L. spinulosa Itô, 1981, and L. weberi Scott A., 1909. However, L. ulleungensis sp. nov. is characterized by the P2 coxa with a reduced inner seta, the P4 exopod second segment without an inner seta, and the anal operculum with a long median projection, a single spine, and a group of outer spines on each side. In a molecular analysis using the mitochondrial cytochrome c oxidase subunit I (COI) and 18S ribosomal RNA (18S rRNA) genes, the inter-specific variation was 22.525–23.102% and 1.325–1.382% of COI and 18S rRNA between the two new species, respectively. A key to the family Longipediidae is provided herein.

Association between Parkinson’s disease and the faecal eukaryotic microbiota

Parkinson’s disease (PD) is one of the most common neurodegenerative disease, and is so far not considered curable. PD patients suffer from several motor and non-motor symptoms, including gastrointestinal dysfunctions and alterations of the enteric nervous system. Constipation and additional intestinal affections can precede the classical motor symptoms by several years. Recently, we reported effects of PD and related medications on the faecal bacterial community of 34 German PD patients and 25 age-matched controls. Here, we used the same collective and analysed the V6 and V7 hypervariable region of PCR-amplified, eukaryotic 18S rRNA genes using an Illumina MiSeq platform. In all, 53% (18) of the PD samples and 72% (18) of the control samples yielded sufficient amplicons for downstream community analyses. The PD samples showed a significantly lower alpha and a different beta eukaryotic diversity than the controls. Most strikingly, we observed a significantly higher relative abundance of sequence affiliated with the Geotrichum genus in the PD samples (39.7%), when compared to the control samples (0.05%). In addition, we observed lower relative abundances of sequences affiliated with Aspergillus/Penicillium, Charophyta/Linum, unidentified Opisthokonta and three genera of minor abundant zooflagellates in the PD samples. Our data add knowledge to the small body of data about the eukaryotic microbiota of PD patients and suggest a potential association of certain gut eukaryotes and PD.

DNA Metabarcoding Methods for the Study of Marine Benthic Meiofauna: A Review

Meiofaunal animals, roughly between 0.045 and 1 mm in size, are ubiquitous and ecologically important inhabitants of benthic marine ecosystems. Their high species richness and rapid response to environmental change make them promising targets for ecological and biomonitoring studies. However, diversity patterns of benthic marine meiofauna remain poorly known due to challenges in species identification using classical morphological methods. DNA metabarcoding is a powerful tool to overcome this limitation. Here, we review DNA metabarcoding approaches used in studies on marine meiobenthos with the aim of facilitating researchers to make informed decisions for the implementation of DNA metabarcoding in meiofaunal biodiversity monitoring. We found that the applied methods vary greatly between researchers and studies, and concluded that further explicit comparisons of protocols are needed to apply DNA metabarcoding as a standard tool for assessing benthic meiofaunal community composition. Key aspects that require additional consideration include: (1) comparability of sample pre-treatment methods; (2) integration of different primers and molecular markers for both the mitochondrial cytochrome c oxidase subunit I (COI) and the nuclear 18S rRNA genes to maximize taxon recovery; (3) precise and standardized description of sampling methods to allow for comparison and replication; and (4) evaluation and testing of bioinformatic pipelines to enhance comparability between studies. By enhancing comparability between the various approaches currently used for the different aspects of the analyses, DNA metabarcoding will improve the long-term integrative potential for surveying and biomonitoring marine benthic meiofauna.

Patterns of Cryptic Diversity and Phylogeography in Four Freshwater Copepod Crustaceans in European Lakes

Comparative phylogeography has become a powerful approach in exploring hidden or cryptic diversity within widespread species and understanding how historical and biogeographical factors shape the modern patterns of their distribution. Most comparative phylogeographic studies so far focus on terrestrial and vertebrate taxa, while aquatic invertebrates (and especially freshwater invertebrates) remain unstudied. In this article, we explore and compare the patterns of molecular diversity and phylogeographic structure of four widespread freshwater copepod crustaceans in European water bodies: the harpacticoids Attheyella crassa, Canthocamptus staphylinus and Nitokra hibernica, and the cyclopoid Eucyclops serrulatus, using sequence data from mtDNA COI and nuclear ITS/18S rRNA genes. The three taxa A. crassa, C. staphylinus and E. serrulatus each consist of deeply diverged clusters and are deemed to represent complexes of species with largely (but not completely) non-overlapping distributions, while in N. hibernica only little differentiation was found, which may however reflect the geographically more restricted sampling. However, the geographical patterns of subdivision differ. The divisions in A. crassa and E. serrulatus follow an east–west pattern in Northern Europe whereas that in C. staphylinus has more of a north–south pattern, with a distinct Fennoscandian clade. The deep mitochondrial splits among populations of A. crassa, C. staphylinus and E. serrulatus (model-corrected distances 26–36%) suggest that divergence of the lineages predate the Pleistocene glaciations. This study provides an insight into cryptic diversity and biogeographic distribution of freshwater copepods.

Deep-Sea Carbonates Are a Reservoir of Fossil Microbes Previously Inhabiting Cold Seeps

Carbonates are globally distributed particularly around deep-sea cold seeps. The embedded microbes are fossil records of the past bioprocess but metagenomes of the carbonates have not been fully studied. In this study, we report microbial community structures and genomes of dominant species in cold-seep carbonates from the South China Sea (SCS) and Gulf of Mexico (GoM). The carbonates contained both anaerobic microbes represented by methane oxidizing archaea (ANME) and aerobic ammonia-oxidizing archaea (AOA). The samples from GoM were mostly composed of small microbial groups, indicating heavy degradation of the fossil microbes. The composition of the carbonate communities differed from that of cold seep sediments, suggesting alteration of cold-seep microbial structures during formation and weathering of carbonates. Extraction of 18S rRNA genes from metagenomic reads revealed prevalence of fungal species in the carbonates of the GoM. Genome binning resulted in 10 genomes for dominant prokaryotic species. The ANME genomes showed a short genetic distance to the relatives from the current cold seep sediments; the AOA genomes were affiliated with alpha ecotype dominating deep-sea sediments. Our study reports the genomes in ancient carbonates and sheds lights on microbial role in formation and bioweathering of carbonates.

Occurrence and Genetic Diversity of Babesia caballi and Theileria equi in Chilean Thoroughbred Racing Horses

This study aimed to serologically and molecularly survey Babesia caballi and Theileria equi in thoroughbred horses from racecourses in Chile. Additionally, the genetic diversity of the positive samples was assessed. A total of 286 thoroughbred horses from the Santiago and Valparaíso racecourses had their serum samples submitted to an ELISA for B. caballi and T. equi, and 457 samples (from the Santiago, Valparaíso, and Concepción racecourses) were tested with nested PCRs for the B. caballi 48 KDa rhoptry protein (RAP-1) and T. equi 18S rRNA genes. Selected RAP-1 and 18S positive products were sequenced to perform phylogenetic and haplotype analyses. An overall seroprevalence of 35.6% was observed for these Chilean racecourses: 23.7% for T. equi, 8.4% for B. caballi, and 3.5% for both agents. Overall, a 53.6% occurrence by nPCR was detected for the three Chilean racecourses: 44.2% for T. equi, 5.4% for B. caballi, and 3.9% for both agents. Phylogenetic analysis of T. equi and B. caballi showed genetic proximity with sequences previously detected in other countries. Haplotype analysis revealed a low diversity among the Chilean sequences, which may have originated from those reported in Brazil, Israel, or Cuba. Babesia caballi and T. equi were detected for the first time in Chilean thoroughbred horses.

A new amphibious species of the genus Scolopendra Linnaeus, 1758 (Scolopendromorpha, Scolopendridae) from the Ryukyu Archipelago and Taiwan

In Japan and Taiwan, five valid species of the genus Scolopendra Linnaeus, 1758 have been described: S. morsitans Linnaeus, 1758, S. subspinipes Leach, 1816, S. mutilans Koch, 1878, S. japonica Koch, 1878, and S. multidens Newport, 1844. Recently, an undetermined species was found in the Ryukyu Archipelago and Taiwan. Using molecular phylogenetic analyses with mitochondrial COI and 16S rRNA and nuclear 28S rRNA and 18S rRNA genes as well as conventional morphological examination, we successfully discriminated this sixth species as an independent lineage from S. subspinipes, S. mutilans, and other named congeners from East and Southeast Asia. Therefore, the species was described as S. alcyona Tsukamoto & Shimano, sp. nov. Several situational evidences suggest that this species prefers streamside environments and exhibits amphibious behavior.

Detection Moleculer Of Putative 18S rRNA Gen Protozoa Trichodina sp. Infected Larvae Gurami (Osphronemus gouramy L) in Balai Benih Ikan Kutasari Purbalingga Central Java

Trichodina spp. are ectoparasitic pathogens of ciliata group that commonly infect both freshwater and marine fish, including gouramy fish. As a result of infection of Trichodina spp. this will lead to inhibition of fish growth and decreased fish production, resulting in low fish selling value. The rate of occurrence of Trichodina spp. that infects gurami can reach 100%. Research has been conducted to determine which one Trichodina spp. Protozoa that infects the gouramy seeds of BBI (Fish Seed Center) Kutasari Purbalingga following detection of 18S RNA gene. Gene detection method used in this research is Polymerase Chain Reaction (PCR) is a technique of DNA synthesis and amplification in vitro. This research is done following these methodes: (1) sampling of Gurami fish with purposive sampling which obtained from BBI Kutasari Purbalingga, (2) isolation of Trichodina spp., (3). Preparation of Trichodina spp. sample and its identification, and (4). Molecular character obervation following detection of 18S rRNA gene. This study obtained 10% percentage of detection of 18S rRNA genes of the species of Trichodina paraheterodentata that infect on the gouramy fish of Purbalingga. The percentage rate of detection of these genes is low when compared with the results of the detection of 18S rRNA Trichodina paraheterodentata gene that infects gouramy fish in Banjarnegara.