Agononida Baba and de Saint Laurent, 1996 （Crustacea： Decapoda： Anomura： Galatheoidea： Munididae） from Chinese waters

The present paper reports five species of squat lobster, genus Agononida Baba and de Saint Laurent, 1996, of which A. squamosa （Henderson, 1885） and A. cf. variabilis （Baba, 1988） were not previously reported in Chinese waters. All the specimens are kept in the Marine Biological Museum collection in the Institute of Oceanology, Chinese Academy of Sciences, Qingdao. To date, there have been l 1 species of this genus recorded from China＇s seas. A key to those species is provided in this paper.     

Galatheid squat lobster species from Chinese waters

Five galatheid squat lobster species belonging to four genera of two galatheid families are reported for the first time from Chinese waters, namely Lauriea simulata Macpherson and Robainas- Barcia, 2013, Phylladiorhynchus ikedai （Miyake and Baba, 1965）, Phylladiorhynchus integrirostris （Dana, 1852）, Babamunida sp., and Raymunida elegantissima （de Man, 1902）. The genera Lauriea Baba, 1971 and Babamunida Cabezas, Macpherson, and Machordom, 2008 have not previously been recorded from Chinese waters.     

First report of the genus Lysippe Malmgren, 1866 (Polychaeta: Ampharetidae) from Chinese waters

The ampharetid genus Lysippe Malmgren, 1866 is reported for the first time from Chinese waters. The identification was based on material deposited in the Marine Biological Museum of the Chinese Academy of Sciences (Qingdao). A new species, Lysippe trichobranchia sp. nov. is described. The new species is widely distributed in shallow waters of the Bohai Gulf, the Yellow Sea and the East China Sea at depths of 10-40 m.     

A preliminary phylogenetic analysis of Luidia (Paxillosida: Luidiidae) from Chinese waters with cytochrome oxidase subunit I (COI) sequences

Luidia Forbes (Paxillosida: Luidiidae) are common soft bottom sea stars with 49 described species. Because of substantial morphological diversity, the taxonomy of the genus is complex and hasn’t been resolved definitely. In order to resolve general taxonomic issues, and determine species boundaries and phylogenetic relationships within the genus Luidia, the sequences of cytochrome oxidase subunit I (COI) gene from 24 specimens of Luidia, belonging to eight taxa in Chinese waters, were studied. Three sequences of two species in genus Luidia from GenBank were used to analyze the phylogenetic relationships. The molecular phylogeny exhibited three main clades, each with strong bootstrap support: Clade A including Luidia quinaria from the Sea of Japan; Clade B including seven nominal species (L. quinaria von Martens, L. yesoensis Goto, L. changi Liu, Liao and Li, L. orientalis Fisher, L. avicularia Fisher, L. longispina Sladen and L. hardwicki Gray) from Chinese waters; and Clade C including L. maculata Müller & Troschel from Chinese waters. Our molecular phylogeny results support the morphological Quinaria-Group and Alternata-Group assigned by Döderlein. Seven nominal species we sampled do not exhibit genetic distances that are large enough to recognize them as separate species. Cryptic species may exist in’ Luidia quinaria’ from the Yellow Sea and the Sea of Japan. Meaningful morphological characters need further investigation in Luidia.     

A new record of a flathead fish (Teleostei: Platycephalidae) from China based on morphological characters and DNA barcoding

A new record of Platycephalus sp.1 (sensu Nakabo, 2002) was documented based on morphological characters and DNA barcoding. We collected 174 specimens of the genus Platycephalus from Chinese coastal waters of Dongying, Qingdao, Zhoushan, and Beihai. Samples were identified as Platycephalus sp.1 morphologically. The coloration, meristic traits, and morphometric measurements are consistent with previously published records. In brief, it is an orange-brown flathead fish with dark brown spots scattered on head and body, lateral line scales 83 to 99 with one or two spine-bearing anteriormost pored scale, no yellow blotch on the caudal fin. Cytochrome oxidase I subunit (COI) gene fragments were sequenced for phylogenetic analysis. The mean evolutionary distance within the species Platycephalus sp.1 was 0.1%. Net evolutionary distances between Platycephalus sp.1 and other species of Platycephalus ranged from 10.8% to 19.7%, which is much greater than the threshold for species delimitation. The COI sequence analysis strongly supports the validity of Platyceohalus sp.1 at genetic level.     

Three new records of deep-water goniasterids (Echinodermata: Asteroidea: Goniasteridae) from China seas

In this paper,three deep-water species of the family Goniasteridae,Ceramaster misakiensis(Goto,1914),Nymphaster arthrocnemis Fisher,1913 and Pontioceramus grandis Fisher,1911,are recorded for the first time from Chinese waters based on collections deposited in the Marine Biological Museum,Chinese Academy of Sciences.The specimens examined were collected during the period 1956 to 1978 from the East China and South China Seas at depths of 184 to 472 m.Diagnosis,detailed figures,and the geographic distributions are provided.A revised list of Goniasteridae recorded from Chinese waters is proposed.     

Molecular phylogeny and evolution of Scomber (Teleostei: Scombridae) based on mitochondrial and nuclear DNA sequences

A molecular phylogenetic analysis of the genus Scomber was conducted based on mitochondrial (COI, Cyt b and control region) and nuclear (5S rDNA) DNA sequence data in multigene perspective. A variety of phylogenetic analytic methods were used to clarify the current taxonomic Classification and to assess phylogenetic relationships and the evolutionary history of this genus. The present study produced a well-resolved phylogeny that strongly supported the monophyly of Scomber. We confirmed that S. japonicus and S. colias were genetically distinct. Although morphologically and ecologically similar to S. colias, the molecular data showed that S. japonicus has a greater molecular affinity with S. australasicus, which conflicts with the traditional taxonomy. This phylogenetic pattern was corroborated by the mtDNA data, but incompletely by the nuclear DNA data. Phylogenetic concordance between the mitochondrial and nuclear DNA regions for the basal nodes Supports an Atlantic origin for Scomber. The present-day geographic ranges of the species were compared with the resultant molecular phylogeny derived from partition Bayesian analyses of the combined data sets to evaluate possible dispersal routes of the genus. The present-day geographic distribution of Scomber species might be best ascribed to multiple dispersal events. In addition, our results suggest that phylogenies derived from multiple genes and long sequences exhibited improved phylogenetic resolution, from which we conclude that the phylogenetic reconstruction is a reliable representation of the evolutionary history of Scomber.     

Age, maturation, and population structure of the Humboldt squid Dosidicus gigas off the Peruvian Exclusive Economic Zones

Age, maturation and population structure of the Humboldt squid Dosidicus gigas were studied based on random sampling of the Chinese jigging fishery off the Peruvian Exclusive Economic Zones (EEZ) during 2008–2010. Estimated ages ranged from 144 to 633 days, confirming that the squid is a short-lived species with longevity no longer than 2 years. Occurrence of mature females and hatching in each month indicated that Humboldt squid spawned year-round. Back-calculated hatching dates for the samples were from January 22^nd, 2008 to April 22^nd, 2010 with a peak between January and March. Two size-based and two hatching date-based populations could be defined from mantle length (ML) at maturity and back-calculated hatching dates, respectively. Females matured at a larger size than males, and there was a significant difference in ML at maturity between the two hatching groups (P <0.05). The waters adjacent to 11°S off the Peruvian EEZ may be a potential spawning ground. This study shows the complexity of the population structure and large variability in key life history parameters in the Humboldt squid off the Peruvian EEZ, which should be considered in the assessment and management of this important resource.     

Designation of a new genus <Emphasis Type="Italic">Bathymenes</Emphasis> for the deep-sea pontoniine shrimps of the ‘<Emphasis Type="Italic">Periclimenes alcocki</Emphasis> species group’ (Decapoda,Caridea, Palaemonidae), with a checklist of the species assigned to the genus

A new genus of the deep-sea pontoniine shrimps, Bathymenes gen. nov., is established for the ‘Periclimenes alcocki species group’ of the genus Periclimenes Costa, 1844. The new genus is distinguished from other genera of the Pontoniinae by a combination of characters: the posteriormost dorsal rostral tooth remote from other teeth, situated in the epigastric position, cornea usually reduced, the propodus of second pereiopods covered with fine granules, the dactylus of the major second chela being generally flanged, the ambulatory pereiopods with the dactyli being biunguiculate and telson with more than two pairs of dorsolateral spines. The genus is mainly distributed in the tropical to warm-temperate Indo-West Pacific waters at depths greater than 200 m. Fifteen species previously recognized as belonging to the ‘P. a lcocki species group’ are now placed in Bathymenes gen. nov. A key for their identification and a checklist of congeneric species are provided.     

Purification and characterization of 2-haloacid dehalogenase from marine bacterium Paracoccus sp. DEH99, isolated from marine sponge Hymeniacidon perlevis

2-haloacid dehalogenases constitute a group of dehalogenases which are capable of dehalogenating the halogenated organic compounds. So far, the 2-haloacid dehalogenases have been found in many bacteria, but not in Paracoccus genus. In the present study, one enzyme 2-haloacid dehalogenase(designated as Deh99), induced by DL-2-chloropropionate(DL-2-CPA), was purified from the marine bacterium Paracoccus sp. DEH99, isolated from marine sponge Hymeniacidon perlevis. The enzyme of Deh99 was purified to homogeneity by ammonium sulfate precipitation, ion exchange chromatography(Q-Sepharose HP), and Superdex 200 gel filtration chromatography. The molecular weight of Deh99 was estimated to be 25.0 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE), and 50.0 kDa natively by gel filtration chromatography. The enzyme of Deh99 stereospecifically dehalogenated L-2-CPA to produce D-lactate, with an apparent Michaelis-Menten constant(Km) value of 0.21 mmol L-1 for L-2-CPA. The optimal pH and temperature for Deh99 activity were 10.0 and 40℃, respectively. The enzyme of Deh99 acted on short-carbon-chain 2-haloacids, with the highest activity towards monochloroacetate. The activity of Deh99 was slightly affected by DTT and EDTA, but strongly inhibited by Cu2+ and Zn2+. The enzyme of Deh99 shows unique substrate specificity and inhibitor sensitivities compared to previously characterized 2-haloacid dehalogenases and is the reported one about purified 2-haloacid dehalogenase isolated from the bacteria of Paracoccus genus.     

Digestive enzyme activities in mudskipper Boleophthalmus pectinirostris and Chinese black sleeper Bostrichthys sinensis

The mudskipper Boleophthalmus pectinirostris and Chinese black sleeper Bostrichthys sinensis occupy the intertidal zone. However, both species have their own unique diet. The former is an herbivore and the latter is a carnivore. In order to reveal the relationship between digestive enzyme activities and diets in the two species, the activities of protease (P), non-specific bile salt-activated lipase (BAL) and α-amylase (A) were determined in the stomach and intestine of adult mudskipper B. pectinirostris and Chinese black sleeper B. sinensis. The results showed that the activities of protease, BAL and α-amylase in the intestine of B. pectinirostris were significantly (P<0.05) higher than those in the stomach. In B. sinensis, gastric protease activity was not different from the intestinal protease (P>0.05), while BAL and α-amylase activities of the intestine were significantly (P<0.05) higher than those of the stomach. The activity of gastric protease in B. sinensis was significantly (P<0.05) higher than that in B. pectinirostris, while the activities of intestinal protease were not different between the two fish species (P>0.05). BAL activities of the stomach and intestine in B. sinensis were significantly (P<0.05) higher than those in B. pectinirostris, while α-amylase activities of the stomach and intestine in B. pectinirostris were significantly (P<0.05) higher than those in B. sinensis. The ratios of P/BAL, A/P and A/BAL of the digestive tract in B. pectinirostris were 1.5, 107.3 and 158.6, respectively; and those in B. sinensis were 0.2, 1.6 and 0.2, respectively. It can be concluded that food digestion in the adult B. pectinirostris is mainly carried out in the intestine, whereas in the adult B. sinensis it is initiated in the stomach and finishes in the intestine. The activities of BAL and α-amylase in B. pectinirostris and B. sinensis are well correlated with their diets. However, a clear-cut correlation between protease activity and diets is not found in these two species.     

Application of otolith shape analysis for stock discrimination and species identification of five goby species （Perciformes： Gobiidae） in the northern Chinese coastal waters

We tested the use of otolith shape analysis to discriminate between species and stocks of five goby species( Ctenotrypauchen chinensis, Odontamblyopus lacepedii, Amblychaeturichthys hexanema, Chaeturichthys stigmatias, and Acanthogobius hasta) found in northern Chinese coastal waters. The five species were well differentiated with high overall classification success using shape indices(83.7%), elliptic Fourier coefficients(98.6%), or the combination of both methods(94.9%). However, shape analysis alone was only moderately successful at discriminating among the four stocks(Liaodong Bay, LD; Bohai Bay, BH; Huanghe(Yellow) River estuary HRE, and Jiaozhou Bay, JZ stocks) of A. hasta(50%–54%) and C. stigmatias(65.7%–75.8%). For these two species, shape analysis was moderately successful at discriminating the HRE or JZ stocks from other stocks, but failed to effectively identify the LD and BH stocks. A large number of otoliths were misclassified between the HRE and JZ stocks, which are geographically well separated. The classification success for stock discrimination was higher using elliptic Fourier coefficients alone(70.2%) or in combination with shape indices(75.8%) than using only shape indices(65.7%) in C. stigmatias whereas there was little difference among the three methods for A. hasta. Our results supported the common belief that otolith shape analysis is generally more effective for interspecific identification than intraspecific discrimination. Moreover, compared with shape indices analysis, Fourier analysis improves classification success during inter- and intra-species discrimination by otolith shape analysis, although this did not necessarily always occur in all fish species.     

Effects of a Potential Autochthonous Probiotic <Emphasis Type="Italic">Bacillus subtilis</Emphasis> 2-1 on the Growth and Intestinal Microbiota of Juvenile Sea Cucumber, <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka

The effects of Bacillus subtilis 2-1 from the intestine of healthy sea cucumber on the growth, digestive enzyme activities and intestinal microbiota of juvenile sea cucumber (Apostichopus japonicus) were determined in the present study. Sea cucumber was fed with Sargassum thunbergii powder supplemented with B. subtilis 2-1 at different concentrations varying among 0 (control), 10⁵, 10⁷, and 10⁹ CFU g^?1 for 8 weeks. Results showed that the growth performance and intestinal amylase and trypsin activities were significantly increased by dietary B. subtilis 2-1 at 10⁹ CFU g^?1 (P < 0.05). However, dietary B. subtilis 2-1 had no significant influence on the lipase activity in sea cucumber (P > 0.05). The polymerase chain reaction denaturing gradient gel electrophoresis and 16S rRNA gene sequencing analysis indicated that dietary B. subtilis 2-1 at 10⁵ and 10⁷ CFU g^?1 inhibited most of the Proteobacteria including those in genus Vibrio. Dietary B. subtilis 2-1 at 10⁹ CFU g^?1 not only decreased the abundance and species of genus Vibrio, but also increased the intensity of genera Psychrobacter and Bacillus. A specific dosage of dietary B. subtilis 2-1 could increase the growth and modulate the intestinal microbiota of sea cucumber; thus it might be a novel probiotic for keeping the health of sea cucumber.     

Two new free-living marine nematode species of the genus Anoplostoma (Anoplostomatidae) from the mangrove habitats of Xiamen Bay,East China Sea

Two new species of free-living marine nematode from mangrove habitats in Xiamen Bay are identified.Anoplostoma tumidum sp.nov.is characterized by relatively short outer labial setae(0.86–1.00 h.d.),long tail(c 7.2–8.9,c' 8.3–10.5),an instinct swollen distal portion of slender spicule(Sc 94–101 μm),and well developed copulatory bursae without bursal papillae.A.tumidum sp.nov.differs from all valid species of genus Anoplostoma in copulatory apparatus of males with a distinct swollen distal portion of spicule,and a relatively obvious constriction of head.A.paraviviparum sp.nov.is characterized by relatively long outer labial setae(1.11–1.22 h.d.),and tail(c 6.6–8.5,c' 8.6–10.2);elongated spicules with distinct knob-like proximal and pointed distal ends(Sc 46–69 μm);distinct strip-like gubernaculum(length with 11–15μm);well developed copulatory bursae with precloacal papillae and post-cloacal papillae;and a distinct constriction of head.A.paraviviparum sp.nov.is similar to A.viviparum Bastian,1865,but differs in the reproductive mode of female and the constriction of head.     

A polysaccharide-degrading marine bacterium Flammeovirga sp. MY04 and its extracellular agarase system

Bacteria of the genus Flammeovirga can digest complex polysaccharides (CPs), but no details have been reported regarding the CP depolymerases of these bacteria. MY04, an agarolytic marine bacterium isolated from coastal sediments, has been identified as a new member of the genus Flammeovirga. The MY04 strain is able to utilize multiple CPs as a sole carbon source and grows well on agarose, mannan, or xylan. This strain produces high concentrations of extracellular proteins (490 mg L-1 ± 18.2 mg L-1 liquid culture) that exhibit efficient and extensive degradation activities on various polysaccharides, especially agarose. These proteins have an activity of 310 U mg-1 ± 9.6 U mg-1 proteins. The extracellular agarase system (EAS) in the crude extracellular enzymes contains at least four agarose depolymerases, which are with molecular masses of approximately 30-70 kDa. The EAS is stable at a wide range of pH values (6.0-11.0), temperatures (0-50℃), and sodium chloride (NaCl) concentrations (0- 0.9 mol L-1). Two major degradation products generated from agarose by the EAS are identified to be neoagarotetraose and neoagarohexaose, suggesting that β-agarases are the major constituents of the MY04 EAS. These results suggest that the Flammeovirga strain MY04 and its polysac-charide-degradation system hold great promise in industrial applications.     

First record of Biecheleria cincta (Dinophyceae) from Chinese coasts, with morphological and molecular characterization of the strains

The presence of Biecheleria cincta (=Woloszynskia cincta) in the Chinese coasts is reported for the first time. In scanning electron microscope, three to five series of vesicles and an elongated apical vesicle (EAV) were visible in the epicone, and both the hypocone and the cingulum had three series of vesicles each. Thin sections revealed that B. cincta possesses stalked pyrenoids and an unusual eyespot consisting of a stack of cisternae with brick-like materials (type E), thus supporting its transfer from Woloszynskia to Biecheleria. Spiny cysts formed spontaneously in culture, with an encystment rate of around 20%. Both large subunit ribosomal DNA (LSU rDNA) and internal transcribed spacer region (ITS) sequences in 12 strains from the Chinese coasts were determined. Phylogenetic analyses based on LSU rDNA and ITS sequences using Bayesian inference and maximum likelihood revealed two distinct ribotypes (referred to as ribotype A and B) in B. cincta. ITS region pairwise distances within B. cincta ranged from 0.024 to 0.072, suggesting the existence of a complex of cryptic species.     

First record and description of male Rhopalione sinensis Markham, 1990, from China,with remarks on genus Rhopalione Pérez, 1920(Crustacea: Isopoda: Bopyridae)

The bopyrid isopod species Rhopalione sinensis Markham, 1990, is recorded for only the second time, from a new locality in China outside Hong Kong, the type-locality. This is also the fi rst fi nding of the male R. sinensis, which is herein described. Its host, Arcotheres sinensis(Shen, 1932) is a new record. A review of the four known species of Rhopalione Pérez, 1920, an updated diagnosis of the genus and a key to species are provided.     

Inhibitory activity of an extract from a marine bacterium Halomonas sp. HSB07 against the red-tide microalga Gymnodinium sp. （Pyrrophyta）

In recent years, red tides occurred frequently in coastal areas worldwide. Various methods based on the use of clay, copper sulfate, and bacteria have been successful in controlling red tides to some extent. As a new defensive agent, marine microorganisms are important sources of compounds with potent inhibitory bioactivities against red-tide microalgae, such as Gymnodinium sp. （Pyrrophyta）. In this study, we isolated a marine bacterium, HSB07, from seawater collected from Hongsha Bay, Sanya, South China Sea. Based on its 16S rRNA gene sequence and biochemical characteristics, the isolated strain HSB07 was identified as a member of the genus Halomonas. A crude ethyl acetate extract of strain HSB07 showed moderate inhibition activity against Gymnodinium sp. in a bioactive prescreening experiment. The extract was further separated into fractions A, B, and C by silica gel column chromatography. Fractions B and C showed strong inhibition activities against Gymnodinium. This is the first report of inhibitory activity of secondary metabolites of a Halomonas bacterium against a red-tide-causing microalga.     

A bio-optical inversion model to retrieve absorption contributions and phytoplankton size structure from total minus water spectral absorption using genetic algorithm

We propose a bio-optical inversion model that retrieves the absorption contributions of phytoplankton and colored detrital matter(CDM),as well as the phytoplankton size classes(PSCs),from total minus water absorption spectra.The model is based on three-component separation of phytoplankton size structure and a genetic algorithm.The model performance was tested on two independent datasets(the NASA bio-Optical Marine Algorithm Dataset(NOMAD) and the northern South China Sea(NSCS) dataset).The relationships between the estimated and measured values were strongly linear,especially for aCDM(412),and the Root Mean Square Error(RMSE) of the CDM exponential slope(SCDM) was relatively low.Next,the inversion model was directly applied to in-situ total minus water absorption spectra determined by an underwater meter during a cruise in September 2008,to retrieve the phytoplankton size structure in the seawater.By comparing the measured and retrieved chlorophyll a concentrations,we demonstrated that total and size-specific chlorophyll a concentrations could be retrieved by the model with relatively high accuracy.Finally,we applied the bio-optical inversion model to investigate changes in phytoplankton size structure induced by an anti-cyclonic eddy in the NSCS.     

Molecular phylogeny of Parapenaeopsis Alcock, 1901 (Decapoda: Penaeidae) based on Chinese materials and 16S rDNA and COI sequence

The phylogenetic relationship among Chinese Parapenaeopsis species was studied by comparing 16S rDNA and COI gene sequence. The results showed that Parapenaeopsis hardwickii (Miers, 1878) and P. cultrirostris Alcock, 1906 identified from Chinese samples were the same species. Parapenaeopsis cornuta (Kishinouye, 1900), P. incisa Liu and Wang, 1987, and P. sinica Liu and Wang, 1987 were different species. However, their systematic relationship were closer than those of other species, and they formed an advanced branch in the phylogenetic trees. Additionally, the systematic position of P. hardwickii (Miers, 1878), P. hungerfordi Alcock, 1905, and P. tenella (Bate, 1888) were not stable in the phylogenetic trees. More genetic markers such as 18S rDNA and 28S rDNA were supposed to be used to confirm the phylogenic relationship of Parapenaeopsis species.