Identification of immune genes in grass carp Ctenopharyngodon idella in response to infection of the parasitic copepod Sinergasilus major

The parasitic copepod Sinergasilus major is an important pathogen of grass carp Ctenopharyngodon idella. To understand the immune response of grass carp to the copepod infection, suppression subtractive hybridization method was employed to characterize genes up-regulation during the copepod infection in liver and gills of the fish. One hundred and twenty-two dot blot positive clones from infected subtracted library were sequenced. Searching available databases by using these nucleotide sequences revealed that 23 genes are immune-related, including known acute-phase reactants, and four novel genes encoding proteins such as source of immunodominant MHC-associated peptides (SIMP), TNF receptor-associated factor 2 binding protein (T2BP), poliovirus receptor-related protein 1 precursor, glycoprotein A repetitions predominant (GARP). The differential expression of seven immune genes, i.e. GARP, alpha-2-macroglobulin, MHC class I, C3, SIMP, T2BP, transferrin, as a result of infection was further confirmed by RT-PCR, with the up-regulation of alpha-2-macroglobulin, MHC class I, C3, SIMP and T2BP in the liver of infected fish, and down-regulation of SIMP in the gills of infected fish. The present study provides foundation for understanding grass carp immune response and candidate genes for further analysis.

Inhibition of the predatory activity of the copepod Mesocyclops notius by Microcystis spp.

The feeding rate of the copepod Mesocyclops notius on two species of cladocerans (Daphnia carinata and Ceriodaphnia cornuta) decreased with increasing environmental concentration of 64-122 mum colonial Microcystis spp. Rate of copepod feeding on Moina micrura was unaffected by the presence of Microcystis spp. Mesocyclops notius rarely preyed on Diaphanosoma brachyurum.

Mortalities induced by the copepod Sinergasilus polycolpus in farmed silver and bighead carp in a reservoir

The frequency distributions of the parasitic copepod Sinergasilus polycolpus were examined in silver carp Hypophthalmichthys molitrix and bighead carp Aristichthys nobilis during a disease outbreak of the 2 species of fish in a reservoir in China. The mean abundance of the copepod was positively related with host length and age, and the overdispersion of the copepod in both silver and bighead carp was fitted well with negative binomial distribution. Although parasite-induced host mortality was observed, a peaked age-parasite abundance curve was not detected in the present parasite-host system. It is also proposed that this peaked age-abundance curve is unlikely to be observed in its natural host populations.

Manipulation of host density affects infestation of a peritrich ciliate (Epistylis lacustris) on a calanoid copepod (Leptodiaptomus minutus) in Crystal Lake, Wisconsin, USA

Specimens of the calanoid copepod, Leptodiaptomus minutus, collected in June 1994 in oligotrophic: north temperate Crystal Lake, were infested with the stalked ciliate Epistylis lacustris. E. lacustris was highly specific to L. minutus and no other coexisting zooplankters were infested. Excluding nauplii, nearly 70% of copepods carried 1-20 ciliates, although the maximum load was as high as 250 ciliates. A lower percentage of nauplii were infested by the ciliate; those that were infested had a lower ciliate load than other copepod stages. Infestation by ciliates had no significant influence on the average egg number of female copepods. In a field experiment, higher copepod densities in enclosures resulted in a significantly higher infestation rate, but the ciliate load per individual copepod did not differ significantly among treatments.

Feeding and egg production of the planktonic copepod Calanus sinicus in spring and autumn in the Yellow Sea, China

Shipboard incubations were conducted in spring (April) and autumn (October/November) 2006 to measure the feeding and egg production rates (EPR) of Calanus sinicus in the Yellow Sea, China. The ingestion rate (2.08-11.46 and 0.26-3.70 mu g C female(-1) day(-1) in spring and autumn, respectively) was positively correlated with microplankton carbon concentrations. In the northern part of the Yellow Sea, feeding on microplankton easily covers the respiratory and production requirements, whereas in the southern part in spring and in the frontal zone in autumn, C. sinicus must ingest alternative food sources. Low ingestion rates, no egg production and the dominance of the fifth copepodite (CV) stage indicated that C. sinicus was in quiescence inside the Yellow Sea Cold Bottom Water (YSCBW) area in autumn. Calanus sinicus ingested ciliates preferentially over other components of the microplankton. The EPR (0.16-12.6 eggs female(-1) day(-1) in spring and 11.4 eggs female(-1) day(-1) at only one station in autumn) increased with ciliate standing stock. Gross growth efficiency (GGE) was 13.4% (3-39%) in spring, which was correlated with the proportion of ciliates in the diet. These results indicate that ciliates have higher nutrient quality than other food items, but the low GGE indicates that the diet of C. sinicus is nutritionally incomplete.

Effects of single and mixed diatom diets on the reproduction of copepod Calanus sinicus

Under laboratory conditions, the potential influence of diatom diets on reproduction of zoo-plankton Calanus sinicus was studied. Four diatom diet ingredients: Skeletonema costatum (SC), Chaetoceros muelleri (CM), Phaeodactylum tricornutum (PT), diatom mixture (MIX) and a control diet: the flagellate Platymonas subordiformis (PS), were used at the same carbon concentrations of 2.0 mu g mL(-1) C. In a period of 17-day laboratory experiment, the effects of these algae diets on egg production and hatching success of the copepod Calanus sinicus were examined. The diets were analyzed for fatty acid content as an indicator of food quality. The results showed that the female survival of all treatments reached more than 80% except PT. Comparing to the initial value, egg production of Calanus sinicus was reduced in diatom diets (PT, CM), but remained in normal level in SC and MIX, indicating that some single diatom diets had a negative effect on the egg production of Calanus sinicus. Feeding with mixed food however can eliminate this negative effect. Among all the treatments, hatching success in filtered seawater was significantly higher than in algal exudates, indicating that not only diatoms but also other phytoplankton in certain concentration can release extracelluar substance that may inhibit eggs from hatching. Fatty acid analysis showed that both egg production rate and hatching success were negatively correlated to the ratio of 20:5 omega 3 and 14:0 in fatty acid composition.

Summer reproduction of the planktonic copepod Calanus sinicus in the Yellow Sea: influences of high surface temperature and cold bottom water

We have observed that Calanus sinicus retreated from neritic areas in the Yellow Sea and concentrated in the Yellow Sea Cold Bottom Water (YSCBW) area in summer. To investigate the summer reproductive strategy of C. sinicus in this situation, effects of high temperature on reproduction and hatching, as well as geographical variation of in situ egg production rate, were studied by onboard incubation in August 2001. Diel vertical migration (DVM) of females was investigated within and outside the YSCBW, respectively. Onboard incubation at 27 degrees C (i.e. surface temperature) resulted in lower fecundities than that at 9.8 and 12 degrees C (i.e. bottom temperature inside and outside the YSCBW) together with decreased hatching rates and increased naupliar malformation. Egg production was more active at stations outside the YSCBW than inside, where chlorophyll-a concentration was also relatively low. Females inside the YSCBW underwent DVM although they rarely entered the surface layer, but DVM was not observed outside the YSCBW. We conclude that surface temperature in summer has deleterious effects on C. sinicus egg production and hatching, and that it cannot reproduce successfully over the whole area. Inside the YSCBW, egg production is depressed by low food availability, while females outside suffer from high temperatures because of strong vertical mixing.

Onshore-offshore variations of copepod community in northern South China Sea

Copepod communities in onshore and offshore waters show a gradient from primarily near shore to primarily oceanic species. Understanding the transition between these communities is fundamental to determining the range of coastal influence. Copepod communities in the northern South China Sea (nSCS) were studied based on samples collected by vertically towing a net in 10 February-6 March (winter) and 26 August-6 September (summer) of 2004. Calanoida species richness, total copepod abundance, Shannon-Weaver diversity index, and onshore-offshore occurrence of dominant species showed obvious change from onshore to offshore waters. Although the offshore stations had lower abundance than the shelf stations, they had more species and larger diversity index. Abundance of some species (groups) with dominance index > 5% (Calanus sinicus, Euchaeta spp., Temora spp., Paracalanus parvus, and Subeucalanus subtenuis) declined from onshore to offshore waters. Warm water species (Pleuromamma abdominalis, P. gracilis, and P. robusta) occurred in offshore waters in both cruises. Station (q-type) cluster analysis in winter and summer separated copepod community into onshore and offshore communities at similar to 40% level of similarity. The two communities were divided at the position of similar to 100-m isobath. In summer, C. sinicus occurred in the upwelling area east of Hainan Island, indicating the presence of an oversummering stock of this species.

Feeding and respiration rates of a planktonic copepod (Calanus sinicus) oversummering in Yellow Sea Cold Bottom Waters

The distribution, feeding and oxygen consumption of Calanus sinicus were studied in August 2001 on a transect across Yellow Sea Cold Bottom Waters (YSCBW) and two additional transects nearby. The distribution of C. sinicus adults and copepodites stage CV appeared to be well correlated with water temperature. They tended to concentrate in the YSCBW (>10,000 ind. m(-2)) to avoid high surface temperature. Gut pigment contents varied from 0.44 to 2.53 ng chlorophyll a equivalents (chl a equiv.) ind.(-1) for adults, and from 0.24 to 2.24 ng chl a equiv. ind.(-1) for CV copepodites. We found no relationship between gut pigment contents and the ambient chl a concentrations. Although the gut evacuation rate constants are consistent with those measured for other copepods, their low gut pigment contents meant an estimated daily herbivorous ingestion of <3% of body carbon in the YSCBW and <10% outside the YSCBW. However, based on estimates of clearance rates, C. sinicus feeds actively whether in the YSCBW or not, so the low ingestion rates probably reflect shortage of food. Oxygen consumption rates of C. sinicus ranged from 0.21 to 0.84 mul O-2 ind.(-1) h(-1), with high rates often associated with high temperature. From the oxygen consumption rates, daily loss of body carbon was estimated to be 4.0-13.7%, which exceeds our estimates of their carbon ingestion rates. C. sinicus was probably not in diapause, either within or outside the YSCBW, but this cold-water layer provides C. sinicus with a refuge to live through the hot, low-food summer.

Spatial distribution of ciliates, copepod nauplii and eggs, Engraulis japonicus post-larvae and microzooplankton herbivorous activity in the Yellow Sea, China

The abundance of anchovy Engraulis japonicus larvae, >20 mum ciliates, copepod eggs and nauplii, and microzooplankton herbivorous activity were studied in the Yellow Sea in June 2000. Anchovy juveniles and larvae were found in only 6 of the 19 stations sampled. The ciliate communities were dominated by 2 species: Laboea strobila and Strombidium compressum. In the surface waters, the abundance of L. strobila ranged between 0 and 560 ind. l(-1). S. compressum only appeared at Stns 15 to 18 (20 to 3300 ind. l(-1)). L. strobila was found mainly in the top 20 m. The abundance of L. strobila was less than 50 ind, l(-1) in waters deeper than 25 m. S, compressum showed subsurface abundance peaks at the salinity abnormality. Tintinnids occurred occasionally with abundance lower than 100 ind. l(-1), The total ciliate abundance fell in the range of 40 to 3420 ind. l(-1). The ciliate biomass in the surface water and the water column ranged between 0,15 and 6.76 mug C l(-1) and 0.4 and 134.4 mg C m(-2), respectively, In the surface waters, the abundance of copepod eggs and nauplii ranged from 0,3 to 3.1 and 1,1 to 15.6 ind, l(-1), respectively. The average abundance of copepod eggs and nauplii in 4 depth (0, 5, 10 and 20 m) fell in the range of 0.2 to 2.8 and 1.0 to 29.4 ind. l(-1), respectively. As a food item of the E. japonicus post-larvae, the abundance of copepod nauplii and eggs appeared to be low. The abundance peaks of ciliate and E, japonicus post-larvae coincided. Although not found in the gut of E, japonicus post-larvae, aloricate ciliates might be ingested by first-feeding anchovy larvae, preventing initial starvation and prolonging the time to irreversible starvation. On the basis of dilution experiments with positive microzooplankton grazing rates, microzooplankton grazed at rates of 0 to 0.61 d(-1). Grazing pressure of microzooplankton on chlorophyll a standing stock (P-i) and potential chlorophyll a primary production (P-p) were 17 to 46% and 35 to 109% d(-1), respectively.

Summertime ciliate and copepod nauplii distributions and micro-zooplankton herbivorous activity in the Laizhou Bay, Bohai Sea, China

The abundance and biomass of ciliated protozoa and copepod nauplii were investigated at 21 grid stations and two anchored stations in the Laizhou Bay, Bohai Sea, China in June 1998. Dilution incubations were carried out to investigate micro-zooplankton grazing pressure at the anchored stations during spring tide and neap tide. The dominant species were Tintinnopsis amoyensis, T. chinglanensis, T. pallida and aloricate ciliates. A total of 13 species of tintinnids were found. The total abundance of ciliates and nauplii ranged from 30 to 2390 ind l(-1) at grid stations. Tintinnopsis amoyensis was the only ciliate found at the anchored stations and in concentrations which varied from 0 to 6700 ind l(-1). The spatial distribution of ciliates was patchy. Tintinnopsis amoyensis and T. pallida were distributed in the Weihe River mouth and Xiaoqinghe River mouth respectively. The aloricate ciliates, T. chinglanensis and Codonellopsis ostenfeldi dominated offshore in sequence. The water mixing process may affect the spatial pattern of the dominant ciliate species. The abundance and biomass of copepod nauplii were in the range of 0-140 ind l(-1) and 0-7 mu g C l(-1) respectively, with the peak appearing at grid station 15. The total biomass of ciliates and copepod nauplii was in the range of 1(.)5-25 mu g C l(-1). Water column biomass of ciliates and nauplii varied from 2(.)37 to 52(.)3 mg C m(-2). At the anchored stations, the phytoplankton growth rates ranged from undetectable to 0 21 d(-1) and micro-zooplankton grazing rates from 0 13 to 0(.)57 d(-1). The grazing pressure of micro-zooplankton were 12 to 43% of the chlorophyll standing stock and 84 to 267% of the chlorophyll (C) 2000 Academic Press.

Effects of different diets on the reproduction and naupliar development of the copepod Acartia bifilosa

The influence of diatoms on the reproduction and naupliar development of Acartia bifilosa was investigated under laboratory conditions, comparing initial in situ values and laboratory-food treatments. Egg production by A. bifilosa was significantly reduced by one diatom diet (Phaeodactylum tricornutum: Pt) and by two non-diatom diets (Platymonas subordiformis: Ps and Nannochloropsis oculata: No). It was less affected by the other diatom diet (Skeletonema costatum: Sc) or by two mixed-food treatments (D-mix and DG-mix), composed of two diatoms (Pt, Sc) and four species (Pt, Sc, Ps and No), respectively. The negative effect of Pt was eliminated when adult copepods were offered mixed-food diets. There were no significant differences between the hatching success values observed in filtered seawater and in algal exudates, indicating that diatoms did not produce active dissolved toxic substances under the different food concentrations tested. The mortality rate of nauplii was higher with Pt than the other diets, suggesting that this diatom species had a negative effect on egg production, hatching success and naupliar survival simultaneously. Compared to other diets, No and Pt were not beneficial food sources for reproduction and for female and larval survival. (c) 2007 Elsevier B.V. All rights reserved.

Autumn net copepod abundance and assemblages in relation to water masses on the continental shelf of the Yellow Sea and East China Sea

Copepod species diversity, abundance and assemblages in relation to water masses over the continental shelf of the Yellow Sea (YS) and East China Sea (ECS) were studied extensively based on the net plankton samples in autumn 2000. Multivariate analysis based on copepod assemblage resulted in recognition of five groups (Groups 1-5) corresponding to the water masses. Groups 1 and 2 delineated from inshore stations with low salinity YS Surface Water, and offshore stations with YS Cold Water in the YS. Group 3 located in the joint area of YS and ECS mainly with Mixed Water. Groups 4 and 5 in the ECS delineated two assemblages mainly from inshore and shallow stations with ECS Mixed Water in the southeastern ECS, and offshore stations along the ECS shelf edge controlled by saline Kuroshio Water. Salinity and temperature were more important in characterizing copepod assemblage of the continental shelf than chlorophyll a. (c) 2005 Elsevier B.V. All rights reserved.