Copepod parasites of a commercial clam (Meretrix meretrix) from Phuket,Thailand

Three species of copepods are reported from hard clams, Meretrix meretrix (L.), obtained from the market in Phuket, Thailand. They are: Conchyliurus bombasticus Reddiah (Clausidiidae), Ostrincola portonoviensis Reddiah (Myicolidae), and Lichomolgus similis Ho & Kim (Lichomolgidae). The first two species are redescribed based on the newly collected material. Conchyliurus fragilis Reddiah is proposed to be relegated to a synonym of C. bombasticus. L. similis is recorded for the first time from the Indian Ocean.     

Isolation and characterization of a virulent Vibrio sp. bacterium from clams (Meretrix meretrix) with mass mortality

MM5 was a bacterial strain isolated from moribund clam (Meretrix meretrix) collected from a farm with mass mortality outbreak. Primary genotypic and phenotypic identification including 16S rDNA sequence analysis, multilocus sequence analysis (MLSA) of four housekeeping genes (gapA, ftsZ, mreB and topA) and biochemical tests suggested that strain MM5 was a Vibrio species closest to but different from Vibrio furnissii. Our previous study indicated that MM5 could induce a high mortality of M. meretrix (Yue et al., 2010). Quantitative challenge test was performed in this study to further evaluate the pathogenic potential of MM5, which showed that at 84 h post-inoculation, the cumulative mortalities of the MM5-injected group were significantly higher than those of control groups (P < 0.05). Cytopathological and histopathological features of the clam infected by MM5 were carried out by transmission electron microscopy (TEM) and Hematoxylin and Eosin (H&E) staining, respectively. Cytopathologically, foci of MM5 were found in hepatocytes of the clam infected by MM5. In addition, cytopathological lesion was detected in foot of infected clam. Histopathologically, MM5 was detected in different tissues of infected clam, including hepatopancreas, mantle and gill. Challenge test combined with pathological features indicated that MM5 was virulent to M. meretrix.     

Bioaccumulation and distribution of mercury in the hard clam,Meretrix lusoria (Bivalvia:Veneidae)

1. Total mercury concentration in whole soft parts of the hard clam, Meretrix lusoria, increased significantly with increasing exposure concentrations. It reached 4.247–7.084 and 9.956–13.643 μg T-Hg/g dry weight in 5 and 50 μg/l Hg mercury solution, respectively, against a background level of 1.824–0.577μg T-Hg/g dry weight.2. Bioconcentration factor was 4–6 times higher in 5 μg/l Hg than that of 50 μg/l Hg. An estimated biological half-life of 20.2 and 18.4 days in 5 and 50 μg/l Hg, respectively, were also obtained after 14 days uptake.3. Accumulation of mercury in tissues of the clams was greater in the gills and viscera than in the mantle, adductor, foot, or hemolymph.4. The amount of mercury present in the gills is related to be a linear relationship with the mercury content of viscera.     

Intracellular Cu/Zn superoxide dismutase (Cu/Zn-SOD) from hard clam Meretrix meretrix: its cDNA cloning, mRNA expression and enzyme activity

Hard clam (Meretrix meretrix) is an economically important bivalve in China. In the present study, a gene coding for an intracellular Cu/Zn-SOD was cloned and characterized from hard clam. The full-length cDNA of this Cu/Zn-SOD (designated as Mm-icCuZn-SOD) consisted of 1,383?bp, with a 462-bp of open reading frame (ORF) encoding 153 amino acids. Several highly conserved motifs, including the Cu/Zn binding sites [H(46), H(48), H(63), and H(119) for Cu binding; H(63), H(71), H(80), and D(83) for Zn binding], an intracellular disulfide bond and two Cu/Zn-SOD signatures were identified in Mm-icCu/Zn-SOD. The deduced amino acid sequence of Mm-icCu/Zn-SOD has a high degree of homology with the Cu/Zn-dependent SODs from other species, indicating that Mm-icCu/Zn-SOD should be a member of the intracellular Cu/Zn-dependent SOD family. Real-time PCR analysis showed that the highest level of Mm-icCu/Zn-SOD expression was in the hepatopancreas, while the lowest level occurred in the hemocytes. Hard clam challenged with Vibrio anguillarum showed a time-dependent increase in Mm-icCu/Zn-SOD expression that reached a maximum level after 6?h. Mm-icCu/Zn-SOD purified as a recombinant protein expressed in E. coli retained a high level of biological activity, 83?% after 10?min incubation at 10–50?°C, and more than 87?% after incubation in buffers with pH values between 2.2 and 10.2. These results indicated that Mm-icCu/Zn-SOD may play an important role in the innate immune system of hard clam.     

文蛤受精及早期胚胎发育过程的细胞学观察

用普通光镜、荧光显微镜技术和石蜡切片技术三种方法,对文蛤卵在受精及早期胚胎发育过程中的外形和核相变化进行了详细观察。结果表明:文蛤成熟未受精卵呈圆球形,直径90.06μm±5.59μm,核相处于第一次成熟分裂中期;精子为鞭毛型,全长48.05μm±1.60μm,头部呈狭茧形,长度为3.06μm±0.17μm;精卵混合后,精子迅速附着于卵子表面,受精后5min-10min,精子进入卵内并明显膨胀,激活卵子启动两次成熟分裂;分别在受精后20min、30min,受精卵完成第一次和第二次成熟分裂,先后排出第一、第二极体;成熟分裂完成之后,精、卵核体积迅速膨胀到最大,核膜重新出现,形成弥散状的雌、雄原核;受精后35min左右,雌、雄原核在卵子中央发生染色体联合,共同排列在纺锤体的赤道板上,形成第一次有丝分裂的中期分裂相;受精后40min-45min,在纺锤丝的牵引下染色体被拉向两极,结果形成2个大小不等的卵裂球;受精后55min-60min,第二次卵裂结束,形成1大3小4个卵裂球,卵裂过程中的核相变化与第一次卵裂基本相同,只是卵裂方向是与第一次卵裂的卵裂沟呈基本垂直的纵裂;受精后80min-90min,第三次卵裂完成,仍为不等全裂,但自此次起开始进行螺旋分裂。此外,实验中也发现了少量的多精入卵、多极分离和天然三倍体等异常现象。     

不同藻类对成熟前文蛤的生长及其生化成分的影响

为了找到适于文蛤(Meretrix meretrix)暂养、育肥的藻类,通过实验观察了不同藻类对成熟前文蛤生长的影响.实验共分为5组,分别单独投喂4种单胞藻,包括海水小球藻(CMorella vulgaris)(A组)、球等边金藻(Isochrysis golbana)(B组)、青岛大扁藻(Platymonas halgolandica)(C组)和牟氏角毛藻(Chaetoceros müelleri)(D组),及混合投喂海水小球藻、球等边金藻与牟氏角毛藻(E组).结果表明,单胞藻无论是单独投喂还是混合投喂,对成熟前文蛤的壳长和体重增长没有显著影响(P>0.05),其常规生化成分如蛋白质、脂肪、总糖及各种脂肪酸含量在某些实验组之间存在显著性差异(P<0.05).E组蛋白质含量(63.57%)最低,与其他组之间存在显著性差异;E组脂肪含量(8.17%)除低于D组(8.64%)外,高于其他3组,但与其他组间无显著性差异;E组总糖含量(7.08%)除低于C组(7.62%)外,高于其他3组,与其他组之间存在显著性差异(P<0.05).亚油酸(C18:2)、亚麻酸(C18:3)和花生四烯酸(C20:4)均为C组最高,含量分别为2.06%、2.28%和6.50%,与其他组之间存在显著性差异.EPA以D组为最高(8.51%),DHA以B组为最高(10.33%).     

Potential role of cathepsin B in the embryonic and larval development of clam Meretrix meretrix

This study was designed to investigate the possible role of Meretrix meretrix cathepsin B (MmeCB) in embryonic and larval development. MmeCB mRNA expression profile was revealed by semi-quantitative RT-PCR. The level of MmeCB mRNA expression was low in trochophore stage but high in pedveliger stage. MmeCB protein expression was detected in the digestive gland, velum, and epidermis along the edges of the shell in D-larvae and pedveligers by immunocytochemistry. In post larvae, MmeCB protein expression was noticed abundant in the digestive gland, whereas a modest expression was identified in the gill filament. The average shell length of larvae hatched from embryos treated with 0.01, 1, and 10?μmol/L Ca074Me (a cathepsin B inhibitor) was significantly shorter than that of control groups. The metamorphosis rates of larvae treated with 0.01 and 1?μmol/L Ca074Me were significantly lower than that of control groups in 4-day larvae, but not in 5-day larvae. Taken together, these results indicated that MmeCB may have stimulatory effects on embryonic development, metamorphosis, and larval growth during M. meretrix larval development.     

The genus Thermocyclops (Crustacea: Copepoda) in Uzbekistan (Central Asia)

Five Thermocyclops species were found in Uzbekistan: T. crassus, T. dybowskii, T. rylovi, T. taihokuensis, and T. vermifer. The last species has been for the first time recorded in the region. The data on morphology, variability and distribution of these species in Uzbekistan are given. A key to determine to the five species is proposed.     

Mass mortality of Sardinella aurita Valenciennes (Pisces, Clupeidae) in Thessaloniki Bay (Macedonia, Greece)

Mass mortality of Sardinella aurita was observed in Thessaloniki Bay during an abrupt temperature fall in February 1991. The mass of dead fish was estimated to be about 79.4 tonnes.     

Two new species of Paracyclops (Copepoda: Cyclopoida,Cyclopidae) from Africa

Paracyclops longispina n. sp. and Paracyclops altissimus n. sp. are described. New characters derived from detailed examination of body and limb ornamentation are used to differentiate them from other Paracyclops species. Paracyclops longispina n. sp. resembles Paracyclops fimbriatus (Fischer, 1853) and Paracyclops imminutus Kiefer, 1929. It differs from the former by the presence of a well-developed spinular row near the base of the inner setae on the antennal coxobasis in both sexes, and from the latter in the structure of the seminal receptacle and the position of the mid-distal spinular row on the posterior surface of the coxa of leg 1. Paracyclops altissimus n. sp. can be distinguished from other members of genus mainly by the structure of the seminal receptacle and leg 5. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ostrincola humesi sp. nov., a myicolid copepod (Poecilostomatoida) parasitic in rock oysters cultured in the Gulf of Thailand

A new species of poecilostomatoid copepod, Ostrincola humesi (Myicolidae), is described, based on specimens collected from the mantle cavity of the rock oyster, Saccostrea cucullata (Born), cultured in the Gulf of Thailand. A close comparison was made between the new species and Ostrincola koe Tanaka with which the new species was previously confused. A key to the nine species of Ostrincola is provided.     

Toxic effects of mercury on the hard clam,Meretrix lusoria,in various salinities

1. The 96-hr lc₅₀ values for juvenile hard clams, Meretrix lusoria, were 328, 392 and 194 μg/l Hg in 10, 20 and 30 ppt salinities at 25 ± 1°C, respectively; for adult hard clams 341 and 140 μg/l Hg in 20 and 30 ppt salinities, respectively.2. Acclimatizing the adult clams to low salinity of 10 ppt lessened the toxicity of mercury. However, juvenile animals appeared to be more sensitive to mercury poisoning after 96 hr exposure in 10 ppt salinity.3. All embryos exposed to 40 μg/l Hg and above died within 30 hr. In the control, 44% of hatched embryos had developed into D-stage larvae, while those exposed to 20 μg/l Hg were still in the trochophore stage. Most of the retarded larvae developed into abnormal forms within 30 hr at 28°C in 15 ppt salinity.4. In order to maintain water quality and protect natural resources, the recommended safe level of mercury is 0.046 (0.039–0.053) μg/l Hg, based on the estimated 30-hr EC₅₀ for the clam embryos, with an application factor of 0.01.     

A parthenogenetic life cycle in a population of Canthocamptus Staphylinus (Copepoda,Harpacticoida)

The harpacticoid copepod Canthocamptus staphylinus (Jurine) was shown to reproduce parthenogenetically in an oligotrophic Finnish lake. The population was univoltine with peaks of egg production in winter and early spring. Young from both peaks became adults in spring and aestivated as cysts. Laboratory experiments suggested that either high temperature (12 °C or more) or long days will inhibit egg production, but that both factors together are needed to induce encystment. Embryonic survival was low, on the average only 60%. The population was limited by low food levels in winter, and it is suggested that food shortage prevents the species from colonizing the profundal zone of oligotrophic lakes.     

On nine Tropodiaptomus-species (Copepoda,Calanoida) from equatorial East Africa

Of the nine species of Tropodiaptomus occurring in Equatorial East Africa, seven are redescribed and figured in great detail. Their relationships are discussed. A primary homonym is eliminated, and types are indicated wherever possible.     

Cloning and characterization of a hsp70 gene from Asiatic hard clam Meretrix meretrix which is involved in the immune response against bacterial infection

In the present study, a 71.43 kDa heat shock protein cDNA was cloned from Asiatic hard clam Meretrix meretrix. The cDNA was 2292 bp, containing an open reading frame (ORF) of 1959 bp, which encodes a protein of 652 amino acids with a theoretical molecular weight of 71.43 kDa and an isoelectric point of 5.32. Based on the amino acid sequence analysis and phylogenetic analysis, this hsp70 cDNA is a member of cytoplasmic hsc70 (constitutive genes) subfamily in the hsp70 family, and is designated as MmeHsc71. Quantitative RT-PCR was carried out to compare the spatial and temporal expression patterns of MmeHsc71 in the mRNA level between control clams and Vibrio parahaemolyticus-infected clams. Spatially, MmeHsc71 mRNA was found in all tested tissues, including foot, hepatopancreas, mantle and gill. MmeHsc71 mRNA expression level in hepatopancreas and gill displayed a significant increase in vibrio-challenged clams at 24h post-infection compared to control clams (P < 0.05). Temporally, there was a significant increase of MmeHsc71 mRNA level in hepathopancreas of vibrio-challenged clams compared to control clams at 6, 12, and 24h post-challenge, respectively. The result of quantitative immunofluorescence also indicated that there was obvious increase of MmeHsc71 in hepatopancreas of vibrio-challenged clams compared to control clams in protein level at 24h post-infection. The results suggested that MmeHsc71 may play an important role in mediating the immune responses of M. meretrix to bacterial challenge.     

A new interpretation of the female genitalia in Macrocyclops albidus (Copepoda, Cyclopidae)

The female genital structures of Macrocyclops albidus (Cyclopidae, Eucyclopinae) were studied using light and electron microscopy. The results confirm that the exterior genital area shows only a copulatory pore, located anteromedially on the ventral face of the genital double-somite, and paired gonopores (not directly visible), situated laterally under the P6 plates. An internal seminal receptacle, composed of several parts, is connected to the gonopores by ventro-lateral cuticular extensions or seminal ducts. The lateral site of communication shows a complex set of connections between the seminal receptacle and the oviducts (via the egg-laying ducts). The structure until now designated as ‘transverse ducts’, visible by transparency on the ventral face, is in fact constituted of internal cuticular thickenings resulting of the fusion of the 6th thoracic somite and the 1st abdominal somite forming the genital double-somite and appearing externally as a part of the suture line; the term ‘suture cord’ is proposed to designate it. The functioning of the system is explained.     

The freshwater Copepoda (Crustacea) of Costa Rica with notes on some species

Sixteen species of Copepoda were recorded in this first detailed study of Costa Rican zooplankton. Two of these species are halophile. Notes are given on some of the species and their distribution. One new species found will be described elsewhere. The Costa Rican fauna resembles in composition the fauna of other Central American and Caribbean regions. Relatively few species were recorded. This is partly due to a low intensity of sampling but also to the paucity of lentic habitats. The occurrence of two circumtropical species of Mesocyclops is interesting.

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Résumé 16 espèces de Copépodes ont été inventoriées au cours d'une première étude détaillée du zooplankton dulcaquicole de Costa Rica. Deux des espèces trouvées sont halophiles. Des remarques sont faites a propos de quelques espèces et de leur répartition. Une nouvelle espèce est nouvelle — elle sera décrite par ailleurs. La fauna de Costa Rica ressemble par sa composition à celle des autres pays d'Amérique centrale et des îles caraibes. Relativement peu d'espèces sont notées. La faible intensité de l'échantillonnage mais aussi la pauvreté en habitats limniques en sont responsables. La présence de deux espèces circumtropicales de Mesocyclops est intéressante.

     

Purification and kinetic characteristics of strombine dehydrogenase from the foot muscle of the hard clam (Meretrix lusoria)

Strombine dehydrogenase (SDH, EC 1.5.1.22) from the foot of the hard clam Meretrix lusoria was purified over 470-fold to apparent homogeneity. It has a monomeric structure with a relative molecular mass of 46,000. Two isoenzymes were identified with isoelectric points of 6.83 and 6.88. SDH is heat labile, and has pH and temperature optima of 7.4–7.6 and 45–46 °C, respectively. l-Alanine, glycine, and pyruvate are the preferred substrates. l-Serine is the third preferred amino acid. Iminodiacetate with the lowest K_i of SDH at both pH 6.5 and 7.5 was the strongest inhibitor among succinate, acetate, iminodiacetate, oxaloacetate, and l-/d-lactate. The inhibitory activities of succinate at pH 6.5, and iminodiacetate and oxaloacetate at pH 7.5 on the SDH were higher. These inhibitors are either competitive or mixed-competitive inhibitors. Half of the enzymatic activity of SDH was inhibited by 0.2 mM Fe³⁺ and 0.6 mM Zn²⁺.