Bacterial response to presence of eukaryote inhibitors in water from a coastal marine environment

A method involving chemical (cycloheximide, thiram) inhibition of protozoan bacterivores was applied in an attempt to estimate marine bacterioplanktonic growth rates. The inhibitors were capable of preventing growth of marine protozoan bacterivores at concentrations which did not inhibit growth of marine bacterial assemblages. Use of the predator-inhibition method with incubated water samples from a nearshore marine environment indicated that protozoan grazing control of the size of bacterial standing crop was quite variable with time and site. The apparent degree of control ranged from non-existent to complete. As a means of estimating bacterial generation times, the predator-inhibition method gave results more closely aligned with those of a dividing-cells technique than a [³H]thymidine technique.     

Determining Water Depths for Monitoring Coastal Water Quality Using Multiple Functional Traits of Periphytic Protozoa in Marine Ecosystems

Ocean Science Journal - Biological trait analysis (BTA) of protozoan communities has been used a useful bioindicator of environmental quality status in marine ecosystems. To determine the optimal...     

纤毛虫诱导鱼类免疫反应的研究

本文综述了纤毛虫诱导鱼类发生免疫反应的类型，免疫反应机制以及纤毛虫疫苗的研究现状。鱼类对纤毛虫诱导产生免疫反应的研究多集中在小瓜虫诱导鱼类免疫反应的研究。鱼类对瓜虫免疫保护机制已基本弄清且已成功研制出小瓜虫疫苗，这是目前唯一的应用于实验室的寄生原生动物疫苗。     

同时检测海洋贝类包纳米虫和派琴虫的双重TaqMan MGB探针实时荧光PCR方法

根据包纳米虫（Bonamia sp.）SSU rDNA和派琴虫（Perkinsus sp.）ITS rDNA的保守区序列,设计特异性引物和Taqman MGB探针,建立了同时检测上述两种贝类原虫的双重实时荧光PCR方法.该方法可检测到包纳米虫基因的446拷贝质粒,以及派琴虫基因的171拷贝质粒,具有较高的灵敏度.以10倍系列稀释的包纳米虫阳性标准品质粒和派琴虫阳性标准品质粒为模板,测得该方法的定量标准曲线相关系数分别为0.999和1.000,显示出很好的扩增效率.同时该方法与尼氏单孢子虫（Haplosporidium nelsoni）、沿岸单孢子虫（H.costale）等其他贝类原虫,以及副溶血性弧菌（Vibrio parahaemolyticus）、迟缓爱德华氏菌（Edwardsiella tarda）和鮰爱德华氏菌（E.ictaluri）等海水养殖常见致病菌之间无交叉反应,表现出较高的特异性.福建莆田的湄州湾、平海湾、兴化湾等地采集的296份贝类临床样本的检测证明,该方法是一种有效的快速检测鉴定上述2种贝类原虫的方法,具有良好的灵敏度和特异性,可广泛应用于海洋贝类原虫感染情况调查,以及贝类疫病检疫监控等领域.     

Protozoa from Blue Lake,Raoul Island (note)

The protozoan populations of microhabitats in Blue Lake, Raoul Island, Kermadec Group, 28° 30’ S 178° 00’ W are described qualitatively and semi‐quantitatively. The in situ and in vitro pH values of the four micro‐habitats examined were recorded. After two days in vitro there was a marked drop in the pH of the mud zones, a slight drop in pH of the water zone and no decrease in the pH at the surface. The nature of the protozoan populations is discussed in relation to the pH values and the changes which took place in these values.     

越冬期鲍养成池中浮游动物密度与种类的变化

研究了杂色鲍(Haliotisdiversicolor)越冬期养殖环境中的浮游动物种类组成和密度变化。共检测到浮游动物33种,其中纤毛虫(Ciliata)23种,鞭毛虫(Mastigophora)5种,轮虫(Rotatoria)4种,桡足类(Copepoda)1种。优势种是奇异猛水蚤(Miraciaefferata)和海洋尾丝虫(Uromenamarinam);钩状狭甲轮虫(Colurellauncinata)和偏体虫(Dysterialancolata)为常见种。砂井水养殖可以减少养殖环境中原生动物以外的各种浮游动物种类。     

Inter-annual variability of the epibiotic community on Pagurus bernhardus from Scotland

Epibiont communities of Pagurus bernhardus and its inhabiting shell collected in the same month of two consecutive years near the Isle of Cumbrae (Scotland) were analyzed. The epibionts found were: (1) the protozoans Acineta, Conchacineta, Corynophrya, Zoothamnium, Cothurnia mobiusi, Cothurnia longipes, Chilodochona, Cryptacineta, Ephelota gemmipara and Ephelota plana; (2) the hydrozoans Leuckartiara, Clytia and Phialella; (3) the entoproct Barentsia; (4) the cirripeds Balanus balanus and Balanus crenatus; (5) the polychaetes Pomatoceros triqueter, Circeis amoricana paguri and Hydroides norvegica. Among these epibionts, a protozoan (E. plana), two hydrozoans (Leuckartiara, and Phialella), and the entoproct (Barentsia) have not been described previously as epibionts on crustaceans. The protozoan, entoproct and hydrozoan epibiont species found on P. bernhardus represent the first mention of their presence on this hermit crab. The protozoan epibionts were found only on the crab, while the hydrozoan epibionts were observed on the shell and on the crab. The cirriped species were observed only on the external surface of the shell, and the polychaete species were observed on the external and internal surfaces of the shell, although Circeis was found only internally. An analysis of the distribution and density of each epibiont species on the anatomical units of the crab, as well as on the different shell areas were made. The comparison between the data of the two years showed differences with respect to the size of crabs, diversity of epibiont groups and density of epibionts. There was a significant difference between the two years with respect to the distribution of epibionts on the anatomical units of the crab, both in terms of density and biomass. The influence of some environmental parameters on these differences is considered.     

Preliminary results of a field study on some stress-related parameters in Tapes philippinarum naturally infected by the protozoan Perkinsus sp.

The aim of the present work was to investigate the possible use of the clam Tapes philippinarum—a filter-feeding bivalve living on soft bottoms—in the monitoring of sediment pollution in the Lagoon of Venice (North-East Italy). To this end, two clam populations, exposed to different environmental conditions (a relative clean site and an industrial polluted area), have been monitored for the prevalence of the protozoan parasite Perkinsus sp. as well as for several biological parameters commonly used as determinants of contaminants exposure. First results showed a low prevalence, higher in the clean site, but a very diffuse distribution of the parasite throughout the two populations. Nevertheless, greater digestive cells height and longer lysosomal latency time were observed in clams from the clean area. As for the physiological stress-related parameters, even if clearance rates were always higher in these clams, scope for growth values indicated a more complicated biological response.     

Spring distribution of larger (>64 μm) protozoans in the Atlantic sector of the Southern Ocean

Distribution of larger protozoans (armoured dinoflagellates, tintinnids, heliozoans, radiolarians and foraminiferans >64 μm) is presented for three major water masses of the Southern Ocean: the Polar Front region (PFr), the southern Antarctic Circumpolar Current (southern ACC) and the northern Weddell Gyre. Sampling took place during the SO-JGOFS cruise ANT X/6 of R/V Polarstern (October–November 1992) along a meridional transect at 6°W between 48°00′S and 59°30′S. Multinet samples (64 μm mesh size) were taken at six stations from the surface down to 500 m depth at five different depth intervals. In the upper 100 m of the water column abundances of larger protozoans varied between 94 and 10,930 ind. m^–3, with highest abundances in the PFr, where phytoplankton blooms occurred, and lowest values in the Antarctic Circumpolar Current–Weddell Gyre Boundary (AWB). Foraminiferans and polycystine and smaller (<300 μm) phaeodarian radiolarians dominated larger protozoan assemblages in the PFr. In open water of the southern ACC, tintinnids, armoured dinoflagellates, foraminiferans and smaller (<300 μm) phaeodarian radiolarians were equally important. The heliozoans Sticholonche spp. and nassellarian radiolarians dominated assemblages in the Weddell Gyre and AWB. Larger protozoan biomasses ranged between 2 and 674 μg C m⁻³ and were always dominated by larger (>300 μm) phaeodarians. Highest biomasses were found in the AWB between 200 and 500 m depth. Standing stocks of larger protozoans constituted a negligible fraction of zooplankton biomass in the upper 200 m of the water column. In deeper layers of the ice-covered Weddell Gyre and AWB their biomasses, dominated by larger (>300 μm) phaeodarians, was significant contributing up to 45% to total larger protozoan and metazoan biomass. Analysis of correlation between distribution patterns and environmental conditions at the stations sampled indicate that spring distribution patterns of heterotrophic armoured dinoflagellates, polycystine radiolarians and foraminiferans follow productivity in the water column. Of the protozoan groups studied the smaller (<300 μm) phaeodarian radiolarians also showed a significant correlation with productivity during spring, however, results from previous studies do not suggest a consistent pattern. Spring distribution patterns of other larger protozoans were not related to differences in productivity in the water column, and effects such as ice-cover, grazing or silica limitation might be determining. Dead radiolarian skeletons constituted on average 27, 8 and 11% of the population of nassellarians, spumellarians and smaller (<300 μm) phaeodarians, respectively. The contribution of dead radiolarian skeletons to total radiolarian stocks varied with depth and water mass. Differences between live and skeleton assemblages composition were observed. These differences should be taken into consideration when interpreting the geological record.     

A numerical analysis of carbon dynamics of the Southern Ocean phytoplankton community: the roles of light and grazing in effecting both sequestration of atmospheric CO2 and food availability to larval krill

Reduced ice extent within coastal regions of the Southern Ocean may lead to deeper surface mixed layers (SML), as prevail in offshore areas. A future decline of ice melt-induced stability of the water column may be associated with a shift in dominant food webs, from larger, sun-adapted diatoms grazed by euphausiids to smaller, shade-adapted flagellates consumed by salps. A basically one-dimensional numerical model of three dominant groups of the Antarctic phytoplankton community (diatoms, cryptophytes, and colonial prymnesiophytes) and four types of herbivore (protozoans, salps, copepods, and euphausiids) is used to explore the seasonal importance of both light limitation and grazing pressure on the amount of annual carbon sequestration and larval krill survival within contrasting oceanic and neritic waters, where respective validation data have been gathered during austral spring by the European JGOFS and RACER programs. With imposition of moderate and large grazing stresses, thought to be typical of offshore waters, we were able to replicate the European JGOFS 1992 observations of light penetration, phytoplankton biomass, primary production, pCO₂, bacterial biomass, labile DOC, ammonium, and total particle effluxes at 100 m within the deep SML of our model. The fidelity of such a large set of simulated state variables suggests that multiple limiting factors are indeed operating on different components of the oceanic phytoplankton community — selective grazing losses on the flagellates, but light limitation of diatoms. Release of protozoan grazing pressure in our model instead leads to unobserved spring blooms of cryptophytes, found only in laboratory enclosures. On an annual basis, weak sequestration of atmospheric CO₂ is simulated in a habitat typical of the Polar Front, while evasion of carbon dioxide occurs under biophysical conditions of the Antarctic Circumpolar Current. Stratification in shallow SML and the same absolute grazing demands by krill and copepods allows sun-adapted diatoms of our model to bloom at the expense of shade-adapted cryptophytes and prymnesiophytes, eaten by salps and protozoans. We were also able to replicate RACER-I observations of the same suite of variables in 1986–1987, as well as the observed 10-fold range of detrital fluxes caught by other sediment trap deployments during 1980 and 1983 along the Antarctic Peninsula. In western Bransfield Strait, coastal waters are a strong sink for atmospheric CO₂ within parcels of Bellingshausen Sea origin, but not perhaps in those from the Weddell Sea, which resemble the oceanic regime of deep SML. We conclude that even in shallow neritic SML, some protozoan rivals of larval krill must still crop flagellates to ensure sufficient abundance of diatom food for both euphausiid survival and possible clogging of the mucous nets of other salp rivals.     

致病性哈维氏弧菌:海洋寄生性纤毛虫的内共生细菌

哈维氏弧菌被观察到与海洋寄生性纤毛虫刺激隐核虫具有内共生现象,其作为一种致病性细菌,可导致感染刺激隐核虫的大黄鱼产生严重的继发性细菌感染。我们通过16s宏测序技术对刺激隐核虫的细菌群落进行鉴定,并通过标准的细菌培养方法分离鉴定出哈维氏弧菌。通过透射电镜和荧光原位杂交方法,在刺激隐核虫的细胞胞质中,均观察到内共生的哈维氏弧菌的存在;而在其细胞核中,则没有相关信号存在。哈维氏弧菌与刺激隐核虫的相关关系以及内共生的哈维氏弧菌对刺激隐核虫的影响作用,仍需要深入进行研究。     

Up-regulation of Hsp60 in response to skeleton eroding band disease but not by algal overgrowth in the scleractinian coral Acropora muricata

Heat shock proteins are biomarkers commonly used to determine the effects of abiotic stresses on the physiology of reef building corals. In this study the effectiveness of the Hsp60 as indicator of biotic stresses in the scleractinian coral Acropora muricata was analyzed, considering the whole holobiont. We focused on two biological interactions recognized to be important contributors to coral reef degradation such as a coral disease, the Skeleton eroding band (SEB) caused by the protozoan Halofolliculina corallasia and the algal overgrowth. In the lagoon of Magoodhoo Island (Maldives) fragments of living tissue of A. muricata exposed to these biotic factors were sampled and proteins subjected to Western analysis. The two different biological interactions trigger diverse responses on Hsp60 level. No detectable effect on Hsp60 modulation appeared in colonies subjected to algal overgrowth. On the contrary, corals displayed a robust up-regulation of Hsp60 in the fragments sampled just above the SEB dark band, where the level of Hsp60 was almost twice compared to the control colonies, indicating that the aggressive behavior of the protozoan causes cellular damage also in coral portions neighboring and along the advancing front of the infection. Portions of coral sampled distant to the SEB band showed a Hsp60 level comparable to that observed in healthy colonies. We propose Hsp60 expression as a promising tool to evaluate physiological stress caused by SEB disease in reef corals.     

桑沟湾养殖生态系统浮游植物的粒径结构及浮游生物对栉孔扇贝碳需求的贡献

The biomass and size fraction of phytoplankton in terms of chlorophyll a(Chl a) was measured during four cruises conducted in April, July, October 2013 and January 2014 in mariculture area, the Sanggou Bay, China.Results show that total Chl a levels in the surface seawater of the Sanggou Bay generally range from 0.10 to 20.46μg/L, with an average value of 2.13 μg/L. Nano-phytoplankton was the most important size-fraction and accounted for about 65.1% of total Chl a. In order to evaluate the importance of the "protozoan trophic link" for energy transfer from the microbial loop to filter-feeding feeders, Zhikong scallop Chlamys farreri was then offered a natural planktonic community as potential prey. Results show that scallops obtained carbon source from natural plankton with the rate of 11 033.05 μg/(g·d). Protists(nanoflagellates and ciliates) were the dominant source of carbon retained by scallop(48.78%). The microbial loop provided 58.45% of the carbon source for farmed scallops. These results indicate that the microbial loop represent a valuable trophic resource in mariculture system of the Sanggou Bay.     

白令海夏季浮游细菌和原生动物生物量及分布特征

1999年7月21日至8月1日在我国首次北极科学考察期间,考察了白令海中部的浮游细菌和原生动物,分析了其丰度、分布、生物量及其生态作用,结果显示,浮游细菌表层生物量为1.5～20.2μg/dm3,平均为浮游植物生物量的30%,100m以上水柱柱总生物量(720～3123mg/m2)平均为浮游植物柱总生物量的67%,因而是白令海夏季与浮游植物处同等量级的优势类群;原生动物表层生物量为1.2～27.4μg/dm3,100m以上水柱柱总生物量为189～1698mg/m2,平均为浮游植物柱总生物量的21%,其中粒径小于5,5～20μm和大于20μm的原生动物分别占其柱总生物量的13%,47%和40%;作为主要类群的异养腰鞭毛虫占原生动物柱总生物量的39%.浮游细菌和原生动物生物量的总体分布趋势从西部向东北和东部递减、从表层向深层衰减,20～25m水层温跃层和表层海流的存在对这一分布特性可能有较大的影响.原生动物受潜在的大、中型浮游动物捕食压力的制约,维持了一个相对较低的生物量水平,在一定程度上限制了微食物环(microbial food loop)在该海域夏季生态系统营养中的作用.     

Protozoa in the diets of Neocalanus spp. in the oceanic subarctic Pacific Ocean

Copepod species of the genus Neocalanus dominate the zooplankton biomass of the oceanic subarctic Pacific Ocean. Neocalanus spp. populations in the subarctic Pacific environment are successful: they feed, accumulate lipid, and persist from year to year. Prior experimental observations derived from a variety of methods indicated that, although their functional morphology is such that they clear the small phytoplankton cells characteristic of the oceanic subarctic Pacific environment efficiently, Neocalanus spp. do not consume sufficient phytoplankton to meet even basic metabolic requirements in that environment. Hence, their success in the subarctic Pacific must depend on their ability to obtain nutrition from other sources. As part of the SUPER (SUbarctic Pacific Ecosystem Research) program, experiments were performed to test the hypothesis that N. plumchrus and N. cristatus obtain a significant portion of their nutrition from planktonic Protozoa. The experiments demonstrate that Protozoa alone do not provide sufficient nutrition for N. cristatus to meet its basic metabolic needs. Protozoa constitute the major dietary component of N. plumchrus however, in agreement with the predictions of Frost's (1987) model of the subarctic Pacific ecosystem. At a minimum this diet permits N. plumchrus to meet basic metabolic requirements. Copepod grazing activities appear to be sufficient to control protozoan stocks in the oceanic subarctic Pacific during late spring and early summer when Neocalanus spp. inhabit the upper water column.     

秋季黄海中华哲水蚤(Calanus sinicus)脂肪酸组成及其指示作用

采用气相色谱法对秋季黄海颗粒悬浮物和中华哲水蚤的脂肪酸组成及其指示作用进行了研究。结果表明,黄海表层海水中硅藻所占比例要大于甲藻。硅藻在黄海南部浮游植物中所占比重高于黄海北部,甲藻在黄海南北部的差别不大。从脂肪酸组成来看,中华哲水蚤雌体、雄体和桡足幼体都摄食硅藻和甲藻,雌体和雄体摄食的硅藻远多于甲藻,而桡足幼体摄食的甲藻则比雌体和雄体多。雄体和桡足幼体植食性的程度高于雌体,而雌体则可能由于繁殖活动的需要,还摄食了原生动物等其它一些营养丰富的食物。对冷水团内外中华哲水蚤脂肪酸组成研究发现,桡足幼体比雌体积累了更多的脂肪酸以满足度夏过程中的能量需求和后期繁殖的需要。     

Cellular responses and disease expression in oysters (Crassostrea virginica) exposed to suspended field contaminated sediments.

Exposure of oysters to water soluble fractions derived from field-contaminated sediments (FCS) containing predominantly lower molecular weight organic aromatic compounds, has been previously demonstrated to enhance pre-existing infections caused by the protozoan parasite, Perkinsus marinus (Dermo), and the prevalence of experimentally induced infections. To further explore the role of pollution on the onset and progression of disease, effects of suspended FCS from an estuarine creek in Virginia, USA, dominated by higher molecular weight polycyclic aromatic hydrocarbons (PAHs) on cellular responses and Dermo disease expression in oysters (Crassostrea virginica) were examined. Sediments were collected from a PAH polluted estuarine creek in Virginia, USA. To test effects on cellular response, oysters from Maine were exposed daily to 0, 1.0, 1.5, or 2.0 g suspended FCS (corresponding to 0, 70.2, 105, or 140 microg PAHs, respectively) for 5, 10, 20, and 40 days. Hemocyte activities and plasma lipid, protein and lactate dehydrogenase (LDH) levels were then measured. Exposure stimulated neutral red uptake, MTT reduction, and 3H-leucine incorporation in oyster hemocytes at various exposure times, but did not affect the plasma protein, lipid and LDH levels. To test effects on Dermo expression, oysters from a Dermo enzootic area, with an initial estimated infection prevalence of 39%, were exposed daily to 0, 1.0, 1.5, or 2.0 g suspended FCS (corresponding to 0, 75.0, 113, or 150 microg PAHs, respectively) for 30 days. Exposure enhanced disease expression in oysters. However, no significant change was noted in any measured cellular or humoral parameters.     

Probing molecular-level transformations of dissolved organic matter: insights on photochemical degradation and protozoan modification of DOM from electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

Molecular-level characterization of natural organic matter (NOM) has been elusive due to the inherent complexity of natural organic mixtures and to the fact that individual components are often polar and macromolecular. Electrospray ionization (ESI) is a “soft” ionization technique that ionizes polar compounds from aqueous solution prior to injection into a mass spectrometer. The highest resolution and mass accuracy of compounds within NOM have been achieved when ESI is combined with an ultrahigh-resolution mass spectrometer such as the Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR MS). With this technique, individual molecules within a variety of natural organic mixtures can be detected and their elemental composition can be determined. At low mass-to-charge (m/z) ratio, the resolution is high enough to assign exact molecular formulas allowing specific components of these mixtures to be identified. In addition to molecular identification, we can now use ESI FT-ICR MS to examine molecular-level changes in different organic mixtures as a function of relevant geo-processes, such as microbial alterations and photochemistry. Here we present the results from the application of ESI FT-ICR MS to two geochemical questions: (1) the effect of photoirradiation on the molecular composition of fulvic acids and (2) the role of protozoan grazers in the modification of DOM in aquatic systems.     

Gill histopathology of two species of buffalo fish from a contaminated Swamp

Two species of buffalo fish, smallmouth buffalo (Ictiobus bubalus) and bigmouth buffalo (Ictiobus cyprinellus), were collected from a contaminated (multiple metals and organic chemicals) Mississippi River Basin ecosystem, Devil's Swamp and a control site, Tunica Swamp, both near Baton Rouge, Louisiana, USA. The buffalo fish were examined for general health and histopathological effects. This research consists of an analysis of the prevalence of infectious and non-infectious disease from the contaminated vs the relatively uncontaminated control site. Tissue burdens were also determined by chemical analysis of buffalo fish muscle (fillets) samples. Both bigmouth buffalo (BMB) and smallmouth buffalo (SMB) from Devil's Swamp accumulated metals in muscle tissue and generally had a higher prevalence of gill lesions than fish collected from the control site, Tunica Swamp. SMB from Devil's Swamp had higher prevalences of lesions than BMB from Devil's Swamp. The primary types of infectious gill lesions observed in the buffalo fish were protozoan infections in the gill filament cartilage and gill lamellae. Chondrolysis, chondritis and bronchitis were associated with parasitic infection. Eosinophils were the predominant cell type involved with bronchitis and chondritis. The proliferative lesions observed were chondroplasia, chloride cell hyperplasia, mucus cell hyperplasia, and epithelial cell hyperplasia and fusion. Based on these preliminary findings, SMB appeared to be a potentially sensitive biomarker of exposure.