First discovery of Perkinsus beihaiensis in Mediterranean mussels (Mytilus galloprovincialis) in Tokyo Bay, Japan.

During analyses of the invasive Mediterranean mussel Mytilus galloprovincialis for pathologies in Tokyo Bay, infection by the protozoan parasite Perkinsus beihaiensis was found through histological examination, Ray's Fluid Thioglycollate Medium assays, and molecular analyses. Specific PCR assays for each Perkinsus species also revealed the presence of an indigenous congeneric species, Perkinsus olseni, but P. beihaiensis was dominant in M. galloprovincialis. Sequences of the ribosomal internal transcribed spacer region I of P. beihaiensis found in Japan were genetically more similar to those found in South American countries (Panama and Brazil) than in Asian countries (China and India). Though Mediterranean mussels have become widespread in Japanese waters since their invasion in the 1930s, epidemiological surveys show that mussels collected outside Tokyo Bay are free of any Perkinsus infections. Based on these results, it was strongly suggested that P. beihaiensis invaded Tokyo Bay by transportation of bivalves originating from South America but has not yet spread to other parts of Japan. The possibility is not ruled out, however, that the parasite is indigenous in Japan but the environment in Tokyo Bay favors its transmission to Mediterranean mussels.

Morphological, Ultrastructural, Motility and Evolutionary Characterization of Two New Hemistasiidae Species.

Until now, Hemistasia phaeocysticola was the only representative of the monogeneric family Hemistasiidae available in culture. Here we describe two new axenized hemistasiids isolated from Tokyo Bay, Japan. Like in other diplonemids, cellular organization of these heterotrophic protists is characterized by a distinct apical papilla, a tubular cytopharynx contiguous with a deep flagellar pocket, and a highly branched mitochondrion with lamellar cristae. Both hemistasiids also bear a prominent digestive vacuole, peripheral lacunae, and paraflagellar rods, are highly motile and exhibit diverse morphologies in culture. We argue that significant differences in molecular phylogenetics and ultrastructure between these new species and H. phaeocysticola are on the generic level. Therefore, we have established two new genera within Hemistasiidae - Artemidia gen. n. and Namystynia gen. n. to accommodate Artemidia motanka, sp. n. and Namystynia karyoxenos, sp. n., respectively. A. motanka permanently carries tubular extrusomes, while in N. karyoxenos, they are present only in starving cells. An additional remarkable feature of the latter species is the presence, in both the cytoplasm and the nucleus, of the endosymbiotic rickettsiid Candidatus Sneabacter namystus.

Microbial Community Dynamics and Assembly Follow Trajectories of an Early-Spring Diatom Bloom in a Semienclosed Bay.

Harmful algal blooms (HABs) are serious ecological disasters in coastal areas, significantly influencing biogeochemical cycles driven by bacteria. The shifts in microbial communities during HABs have been widely investigated, but the assembly mechanisms of microbial communities during HABs are poorly understood. Here, using 16S rRNA gene amplicon sequencing, we analyzed the microbial communities during an early-spring diatom bloom, in order to investigate the dynamics of microbial assembly processes. Rhodobacteraceae, Flavobacteriaceae, and Microbacteriaceae were the main bacterial families during the bloom. The 30 most abundant operational taxonomic units (OTUs) segregated into 4 clusters according to specific bloom stages, exhibiting clear successional patterns during the bloom process. The succession of microbial communities correlated with changes in the dynamics of algal species. Based on the ß-nearest taxon distance, we constructed a simulation model, which demonstrated that the assembly of microbial communities shifted from strong heterogenous selection in the early stage of the bloom to stochasticity in the middle stage and then to strong homogeneous selection in the late and after-bloom stages. These successions were driven mainly by chlorophyll a contents, which were affected mainly by Skeletonema costatum Moreover, functional prediction of microbial communities showed that microbial metabolic functions were significantly related to nitrogen metabolism. In summary, our results clearly suggested a dominant role of determinacy in microbial community assembly in HABs and will facilitate deeper understanding of the ecological processes shaping microbial communities during the algal bloom process.IMPORTANCE Harmful algal blooms (HABs) significantly influence biogeochemical cycles driven by bacteria. The shifts in microbial communities during HABs have been studied intensively, but the assembly mechanisms of microbial communities during HABs are poorly understood, with limited investigation of the balance of deterministic and stochastic processes in shaping microbial communities in HABs. In this study, the dynamics and assembly of microbial communities in an early-spring diatom bloom process were investigated. Our data both confirm previously observed general microbial successional patterns and show new detailed mechanisms for microbial assembly in HABs. These results will facilitate deeper understanding of the ecological processes shaping microbial communities in HABs. In addition, predictions of metabolic potential in this study will facilitate understanding of the influence of HABs on nitrogen metabolism in marine environments.

Two species of Neometadena Hafeezullah & Siddiqi, 1970 (Digenea: Cryptogonimidae) from Moreton Bay, Australia, including the description of Neometadena paucispina n. sp. from Australian Lutjanidae.

A survey of the trematode fauna of lutjanid fishes off the east coast of Queensland (QLD), Australia revealed the presence of two species of Neometadena Hafeezullah & Siddiqi, 1970 (Digenea: Cryptogonimidae). Neometadena paucispina n. sp. is described from the intestine and pyloric caeca of Lutjanus fulviflamma (Forsskål) and L. russellii (Bleeker) from Moreton Bay, in southeast QLD. Specimens of the type- and only other species, N. ovata (Yamaguti, 1952) Miller & Cribb, 2008, were recovered from L. carponotatus (Richardson), L. fulviflamma, L. fulvus (Forster), L. russellii, and L. vitta (Quoy & Gaimard) off Lizard Island, on the northern Great Barrier Reef (GBR). Neometadena paucispina is distinguished from N. ovata in having fewer oral spines (55-65 vs 67-80). Alignment of novel molecular data for these two taxa revealed that they differ consistently by 13 nucleotides (1.5%) over the partial large subunit (LSU), 34 nucleotides (6.6%) over the internal transcribed spacer 1 (ITS1), 0 nucleotides over the 5.8S, and 21 nucleotides (7.3%) over the ITS2 rDNA regions. Despite relatively large samples of L. carponotatus, L. fulviflamma and L. russellii from three distinct locations along the east coast of QLD (i.e. Moreton Bay in the south, Heron Island in central QLD and Lizard Island in northern QLD), these two species have been found at only one site each with neither species at Heron Island. These distributions are discussed in the context of the wide distribution of other cryptogonomid species in the same hosts elsewhere in the Indo-West Pacific.

Clydonella sawyeri n. sp. (Amoebozoa, Vannellida): Morphological and molecular study and a re-definition of the genus Clydonella Sawyer, 1975.

We isolated and described a brackish-water amoeba, Clydonella sawyeri n. sp. (Amoebozoa, Vannellida), from the littoral habitat in Kandalaksha Bay (The White Sea, northwestern Russia). Morphology of this amoeba corresponds to the initially proposed diagnosis of the genus Clydonella Sawyer, 1975, although it is sufficiently different from other described species of this genus to warrant a distinct species designation. Phylogenetic analysis based on the small-subunit rRNA gene shows that this species is closely related to the two previously studied ATCC strains identified as Clydonella sp., for which only ultrastructural and molecular data were published. Cytochrome oxidase subunit 1 (COI) gene sequence of the studied species was for the first time obtained for Clydonella. Analysis of this marker shows that this genus belongs to Vannellida and is separated from the other vannellids sampled to date. Phylogenetic analysis of the concatenated SSU rRNA and COI genes dataset yields the best resolved position of Clydonella compared to both markers when analysed separately. Based on the data presented we finally link light microscopic, ultrastructural and molecular data in a description of a single strain, which allows a refinement of the diagnosis of the genus Clydonella.

[Causes of succession of planktonic algae in Shennong bay of Three Gorges Reservoir in spring in 2014].

Objective: Algal blooms occurred in some sections of Shennong bay after impounding of Three Gorges Reservoir. Methods: Related environmental and hydrodynamic factors were monitored during the period of algal blooming season in 2014 (March 20, April 13, May 23) in Shennong bay, Three Gorges Reservoir. To study succession of planktonic algae, water stable coefficient, euphotic depth and mixed layer depth were used to analyze stratification and hydrodynamic characteristics. Results: We identified 6 phyla, 38 species (genera) planktonic algae. The sensitive area of algal bloom was at SN05 (677.677×105 cells/L) and SN06 (716.761×105 cells/L), and the planktonic algae biomass during this period was significantly different (ANOVA, p<0.05). Moderate water temperature, adequate nutrients, weak stratification and poor mixing promoted the rapid growth and breakout of the diatom bloom with Cyclotella spp. as the dominant species in March. Further increase of water temperature, stronger stratification and decrease of dissolved silicate and mixing layer restricted the diatom growth. Chlorella spp. and Chlamydomonas spp. grew better in shallow mixed layer with rich nutrients and evident stratification. Then Chlorophycean bloom broke out with Chlorella spp. as the dominant species and Chlamydomonas spp. the next-dominant species. High biomass maintained in April. In May, algal bloom gradually vanished due to sharp fluctuation of water level and increase of velocity. Monitored maximum water velocity was 0.1141 m/s at 2 m depth, exceeded an optimal flow rate perfect for growth of planktonic algae. Conclusion: Stratification and hydrodynamic characteristics had important effect on planktonic algae under the condition of adequate nutrients. Velocity became the main factor that inhibited the growth of algae in Shennong bay in pre-flood falling stage of the Three Gorges Reservoir.

Historical trends in the species inventory of tintinnids (ciliates of the microzooplankton) in the Bay of Villefranche (NW Mediterranean Sea): Shifting baselines.

In the Bay of Villefranche, tintinnid ciliates have been studied since 1879 yielding a unique time series. The species inventory, excluding likely synonymous records, numbers 108. Temporal increases in the inventory appear linearly related to sampling effort up until the 2000s with a cumulative sampling effort of about 200 dates. Subsequently, with a large increase in sampling to currently over 460 dates, the rate of increases in species numbers declines. Surprisingly, the inventory is not highly inflated by unique occurrences, as species found but once are only 17 out of the 108. However, in recent years many previously recorded taxa have not been seen. Missing from a species list derived solely from intensive sampling from 2013 to 2016 are 38 previously recorded species. Most (26 out of 38) were recorded from a single year and thus may have been temporary residents. However, 12 species were found in multiple years by different investigators suggesting relatively common occurrence in the past. The substantial effort required to adequately sample a locality and possible historical changes in species inventories suggest that claims of a species as 'new' to a locality should be made with caution & caveats.

Unusually abundant and large ciliate xenomas in oysters, Crassostrea virginica, from Great Bay, New Hampshire, USA.

During routine histological examination of oysters (Crassostrea virginica) from Great Bay, New Hampshire, USA, a high prevalence and intensity of ciliate xenomas has been noted since sampling began in 1997. Xenomas are hypertrophic lesions on the gills of bivalve molluscs caused by intracellular ciliates, likely Sphenophrya sp. Although not known to cause mortality in oysters, xenomas have not previously been reported at this high abundance. The objectives of this study were to characterize the xenomas, describe the ciliates, and gather baseline epizootiological data with correlations to environmental and biological parameters. Upon gross examination, xenomas appeared as white nodules, up to 3mm in diameter, located in the gill tissue and occasionally fusing into large masses along the gill filaments. Light microscopy of histological sections revealed xenomas located in the gill water tubes, which they often completely blocked. Higher magnification revealed dual nuclei, eight kineties, and conjugation of the ciliates. Transmission electron microscopy revealed dual nuclei that varied in density, a maximum of twenty cilia in each kinety radiating from the oral apparatus to the posterior, and a 9+2 axoneme structure within the cilia. These traits place the ciliates into the Order Rhynchodida, but insufficient molecular data exist to confirm classification of this ciliate to the Genus Sphenophrya. Since 1997, xenoma prevalence has fluctuated with peaks in 2000, 2004, and 2011. Infected oysters generally contained <30 xenomas, but 2.1% contained >100, sharply contrasting the rare prevalence and low intensity reported elsewhere. Prevalence increased with oyster size, leveling off near 50% in oysters >60mm. Infection intensity peaked in 70-90mm oysters and declined in larger oysters. Individual oyster condition was not associated with xenoma intensity, but sites with oysters in higher condition generally had a greater prevalence and intensity of xenoma infections. Seasonal data indicated an infection cycle increasing from summer to fall, peaking at 55-65% in November and dropping to <10% by spring. The oyster population in Great Bay, NH warrants further examination to understand the mechanisms and conditions controlling xenoma formation.

Parasite fauna of Epinephelus coioides (Hamilton, 1822) (Epinephelidae) as environmental indicator under heavily polluted conditions in Jakarta Bay, Indonesia.

The objective of this study was to assess the environmental conditions of a heavily polluted marine habitat using descriptors of fish parasites. Epinephelus coioides from Jakarta Bay as well as off Jakarta Bay was studied for metazoan parasites. Based on 70 fish and considering previous studies (230 fish), an environmental indicator system was designed. Including the recent study, a total of 51 parasite species have been recorded for E. coioides in Indonesian waters. Seven of them combined with five parasitological indices are useful descriptors for the environmental status of marine ecosystems. The results are visualized in a star graph. A significant different parasite infection between nine analyzed localities demonstrates the negative influence of the megacity Jakarta onto the coastal environment. We herewith complete a parasite based indicator system for Indonesian coastal waters, and suggest that it can be used in other marine habitats as well as for further epinephelids.

Heterogeneous distribution in sediments and dispersal in waters of Alexandrium minutum in a semi-enclosed coastal ecosystem.

Within the framework of research aimed at using genetic methods to evaluate harmful species distribution and their impact on coastal ecosystems, a portion of the ITS1rDNA of Alexandrium minutum was amplified by real-time PCR from DNA extracts of superficial (1-3cm) sediments of 30 subtidal and intertidal stations of the Bay of Brest (Brittany, France), during the winters of 2013 and 2015. Cell germinations and rDNA amplifications of A. minutum were obtained for sediments of all sampled stations, demonstrating that the whole bay is currently contaminated by this toxic species. Coherent estimations of ITS1rDNA copy numbers were obtained for the two sampling cruises, supporting the hypothesis of regular accumulation of A. minutum resting stages in the south-eastern, more confined embayments of the study area, where fine-muddy sediments are also more abundant. Higher ITS1rDNA copy numbers were detected in sediments of areas where blooms have been seasonally detected since 2012. This result suggests that specific genetic material estimations in superficial sediments of the bay may be a proxy of the cyst banks of A. minutum. The simulation of particle trajectory analyses by a Lagrangian physical model showed that blooms occurring in the south-eastern part of the bay are disconnected from those of the north-eastern zone. The heterogeneous distribution of A. minutum inferred from both water and sediment suggests the existence of potential barriers for the dispersal of this species in the Bay of Brest and encourages finer analyses at the population level for this species within semi-enclosed coastal ecosystems.

Disease Risk & Landscape Attributes of Tick-Borne Borrelia Pathogens in the San Francisco Bay Area, California.

Habitat heterogeneity influences pathogen ecology by affecting vector abundance and the reservoir host communities. We investigated spatial patterns of disease risk for two human pathogens in the Borrelia genus-B. burgdorferi and B. miyamotoi-that are transmitted by the western black-legged tick, Ixodes pacificus. We collected ticks (349 nymphs, 273 adults) at 20 sites in the San Francisco Bay Area, California, USA. Tick abundance, pathogen prevalence and density of infected nymphs varied widely across sites and habitat type, though nymphal western black-legged ticks were more frequently found, and were more abundant in coast live oak forest and desert/semi-desert scrub (dominated by California sagebrush) habitats. We observed Borrelia infections in ticks at all sites where we able to collect >10 ticks. The recently recognized human pathogen, B. miyamotoi, was observed at a higher prevalence (13/349 nymphs = 3.7%, 95% CI = 2.0-6.3; 5/273 adults = 1.8%, 95% CI = 0.6-4.2) than recent studies from nearby locations (Alameda County, east of the San Francisco Bay), demonstrating that tick-borne disease risk and ecology can vary substantially at small geographic scales, with consequences for public health and disease diagnosis.

Gonad-infecting species of Philometra (Nematoda: Philometridae) from groupers Epinephelus spp. (Osteichthyes: Serranidae) in the Bay of Bengal, India.

Based on light and scanning electron microscopical studies, two new and one specifically not identified gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) are described from the ovary of marine fishes of the genus Epinephelus Bloch (Serranidae, Perciformes) in the Bay of Bengal, off the eastern coast of India: P. indica sp. nov. (male and females) from the honeycomb grouper E. merra Bloch, P. tropica sp. nov. (males and females) from the duskytail grouper E. bleekeri (Vaillant) and Philometra sp. (only females) from the cloudy grouper E. erythrurus (Valenciennes). Philometra indica is mainly characterized by the length of spicules 192-195 µm and the gubernaculum 84 µm, the distal tip of the gubernaculum without a dorsal protuberance, and by the presence of five pairs of caudal papillae. Philometra tropica is mainly characterized by the spicules conspicuously ventrally distended at their posterior halves, the distal tip of the gubernaculum with a dorsal protuberance, and the presence of three pairs of caudal papillae.

A quantitative assessment of the role of the parasite Amoebophrya in the termination of Alexandrium fundyense blooms within a small coastal embayment.

Parasitic dinoflagellates of the genus Amoebophrya infect free-living dinoflagellates, some of which can cause harmful algal blooms (HABs). High prevalence of Amoebophrya spp. has been linked to the decline of some HABs in marine systems. The objective of this study was to evaluate the impact of Amoebophrya spp. on the dynamics of dinoflagellate blooms in Salt Pond (MA, USA), particularly the harmful species Alexandrium fundyense. The abundance of Amoebophrya life stages was estimated 3-7 days per week through the full duration of an annual A. fundyense bloom using fluorescence in situ hybridization coupled with tyramide signal amplification (FISH-TSA). More than 20 potential hosts were recorded including Dinophysis spp., Protoperidinium spp. and Gonyaulax spp., but the only dinoflagellate cells infected by Amoebophrya spp. during the sampling period were A. fundyense. Maximum A. fundyense concentration co-occurred with an increase of infected hosts, followed by a massive release of Amoebophrya dinospores in the water column. On average, Amoebophrya spp. infected and killed ∼30% of the A. fundyense population per day in the end phase of the bloom. The decline of the host A. fundyense population coincided with a dramatic life-cycle transition from vegetative division to sexual fusion. This transition occurred after maximum infected host concentrations and before peak infection percentages were observed, suggesting that most A. fundyense escaped parasite infection through sexual fusion. The results of this work highlight the importance of high frequency sampling of both parasite and host populations to accurately assess the impact of parasites on natural plankton assemblages.