Size-selective grazing of coastal bacterioplankton by natural assemblages of pigmented flagellates,colorless flagellates,and ciliates

Fluorescently-labelled bacteria (FLB) were used to study the feeding strategies of a natural assemblage of estuarine protozoans and to examine whether the protozoan grazing could account for the in situ size structure of the bacterioplankton. The FLB, DTAF-stained enterococci, ranging in volume from 0.01 to 0.30 × 10^–1 µm³, were added to a natural planktonic assemblage at a density of 5.5% of the natural bacterioplankton. Initial densities (individuals ml^–1) were as follows: total natural bacteria, 2.2 × 10⁶; FLB, 1.2 × 10⁵; pigmented flagellates, 300; colorless flagellates, 250; and ciliates, 30. FLB consumption rates were determined by examining the contents of protozoan food vacuoles, and the long-term effect of grazing (over a period of 100 hours) was determined by monitoring the decline in the FLB density in experimental vessels. The average consumption rates of FLB by pigmented flagellates were similar to those by flagellates that lacked chloroplasts (0.9 and 0.6 FLB protozoan^–1 hour^–1, respectively). The ciliates consumed bacteria at an average rate that was 17-fold higher (per cell) than flagellates, and they displayed a greater preference for larger bacteria than did the flagellates. FLB of the mid-size classes (0.025–0.100 µm³) were heavily grazed by the entire protozoan assemblage; the smallest (<0.025 µm³) and the largest (>0.100 µm³) FLB escaped protozoan grazing. This had a profound effect on the resulting size distribution of FLB. At the end of a 100-hour incubation, the percentage of mid-size FLB (0.025 to 0.100 µm³) decreased 2.0–2.2-fold, while the percentage of the smallest and the largest FLB increased 2.0–2.5-fold. Resultant densities of FLB were consistent with initial clearance rates determined for the protozoan groups. The grazing rates of protozoans on FLB were species-specific; whereas some species consumed FLB, others did not demonstrate bacterivory. The results suggest that protozoan grazing has a major effect on the size distribution of coastal bacterioplankton. By selectively feeding on a particular size-class of bacteria, planktonic ciliates may consume 15–90% day^–1 of the standing stock of largest size classes of bacterioplankton. Thus, ciliates, which were present in low abundance in the field, could not balance the production of the entire bacterial community, but they may strongly influence the portion of the bacterial community represented by the largest bacterial class. The direct effect of flagellates (e.g., grazing) was limited to smaller bacteria.Offprint requests to: M. P. Shiaris.     

Size-selective grazing of bacteria by Bosmina longirostris--an image-analysis study

Image analyses on the filtering apparatus of Bosmina longirostricshowed that the filter mesh is finer on the gnathobasic filterplates of the second and third trunk limbs (ranges from 0.43to 0.97 µm) and coarser for the outer ones of the thirdlimb (ranges from 0.5 to 1.36 µm), and the intersetulardistances increase with body length. Grazing experiments combinedwith image analysis confirmed the efficient grazing of B.longirostrison natural bacteria with cell lengths equal to or larger thanthe intersetular distances of the gnathobasic filter plates.During the experiments, the animals minimized the average celllength of the bacterioplankton assemblages from 0.77–0.96µm to 0.55–0.68 µm, corresponding to the meanof the filter mesh size on the fine gnathobasic filter platesof the experimental populations. The clearance rates for large,elongated or dividing cells with maximal lengths of 0.88–8.40µm were 2.3–17.7 times higher than those for smallsingle coccoids with a diameter of <0.45 µm. The resultsprovide evidence of a significant differential impact of B.longirostrison the bacterial community structure with respect to the shapeand size of the cells, and demonstrate that the species is amore effective bacterial feeder than considered previously.     

Relationship between the flagellates and the ciliates.

The flagellates and the ciliates have long been considered to be closely related because of their unicellular nature and the similarity in the structures of the axoneme of the flagella and cilia in both groups. Most protozoologists believe that the ciliates arose from a flagellate. The flagellates that are most similar in structure to the ciliates are the dinoflagellates and two genera of uncertain taxonomic position, Colponema and Katablepharis. Structurally, dinoflagellates have a number of similarities with ciliates. These include the similarity of the cortical alveoli in the ciliates to the thecal vesicles in the dinoflagellates, the possession of tubular cristae, the similarity of the parasomal sac of the ciliates to the pusule of the dinoflagellates, the possession of similar trichocysts and mucocysts, and some similarity in the feeding apparatus. Colponema spp. are probably related to the dinoflagellates and have many of the same similarities with the ciliates. Katablepharis spp. are very similar in structure to the swarmer (embryo) of the suctorian ciliates. Indeed, reduction in the number of cilia to two in the suctorian swarmer and elimination of the macronucleus would result in a cell that is very similar to the Katablepharis cell. The feeding apparatus of Katablepharis spp. and the rest of the ciliates consists of two concentric microtubular arrays associated with vesicles. Information available from nucleotide sequencing of rRNA places the dinoflagellates in an ancestral position to the ciliates. The rRNA of Colponema and Katablepharis spp. has not yet been investigated. The use of stop codons in mRNA is discussed in relation to phylogeny.     

Food selectivity and grazing impact on toxic Dinophysis spp. by copepods feeding on natural plankton assemblages

Food selectivity and grazing impact by Acartia bifilosa, Temora longicornis and Centropages typicus on Dinophysis spp. plankton assemblages were experimentally investigated in the Baltic Sea. Toxin analyses were carried out on phyto- and zooplankton-dominated size fractions from field-collected samples to assess if toxins produced by Dinophysis spp. would end up in the zooplankton. All copepod species fed actively on toxic Dinophysis spp. (Dinophysis acuta and Dinophysis norvegica). Despite the non-selective feeding behaviour by T. longicornis and C. typicus, selectivity coefficients on D. acuta progressively decreased as food availability increased. Similar response was not observed for A. bifilosa, which displayed an even less selective behaviour. A. bifilosa had no significant negative effect on the net growth of D. norvegica at the lowest food concentration offered, whereas T. longicornis and C. typicus had significant negative effects on the net growth of D. acuta at low concentrations, similar to those observed in situ. Both species could potentially contribute as a substantial loss factor for Dinophysis spp. provided they are abundant at the onset of the blooms. The estimated grazing impact by the copepod populations was only considerable when C. typicus abundance was high and D. acuta population in sharp decline. Our results suggest that when high abundance of grazers and poor growth condition of prey populations prevail, grazing impact by copepods can contribute considerably to prevent Dinophysis spp. populations to grow or to cause the populations to decline. Okadaic acid was detected in the zooplankton size fraction at one occasion, but the concentration was far lower than the one expected from the ingested toxins. Thus, even if copepods may act as vectors of DSP-toxins to higher trophic levels, the amount of these toxins transported in the food web by copepods seems limited.     

黑色素染色法在纤毛虫和鞭毛虫研究中的使用与改进

介绍并分析了前人在纤毛虫研究中使用过的黑色素染色法,得出作者自己的改进法及其复染法。报告了黑色素改进法的步骤及操作细节,并示出此法显示纤毛虫和鞭毛虫的效果。     

Observations on endonuclear bacteria in euglenoid flagellates

Summary Rod-shaped inclusions in the nuclei and/or cytoplasm of strains ofEuglena spirogyra, Lepocinclis ovum, Strombomonas conspersa andTrachelomonas oblonga var.punctata have been identified as living populations of bacteria by reference to size, shape and ultrastructure. Most strains contain 20–200 bacteria per nucleus. Each bacterial cell is surrounded by a mucilaginous sheath and consists of a double-membraned envelope, dense cytoplasm with small ribosomes, and a filamentous DNA-containing centre. The bacteria multiply to keep pace with euglenoid division but there is no evidence that the euglenoid cells benefit from or are harmed by the association. Attempts to transfer the endonuclear bacteria to uninfected strains and species have been unsuccessful, as have attempts to isolate the bacterium into pure culture. The infected strain ofEuglena spirogyra cannot fix nitrogen. The possible nature of the euglenoid/bacterium association is discussed.Dedicated to Prof. Dr.Lothar Geitler on the occasion of his 70th birthday.     

Bacterivory of metazooplankton, ciliates and flagellates in a newly flooded reservoir

Bacterial consumption by metazoan zooplankton and phagotrophic protists was measured in situ during the period of thermal stratification in the epilimnion (1 m) and metalimnion (7 m) of a newly flooded reservoir (Sep reservoir, France). The mean bacterial consumption was 2.53 x 10⁶ bacterial l^-1 h^-1 at 1m and 0.97 x 10⁶ bacteria l^-1 h^-1 at 7m. The main consumers over the whole study period were the cladocerans Daphnia longispina and Ceriodaphnia quadrangula, accounting on average for 72% of the potential total predation of bacteria at 1 m and 56% at m, especially during the months of May-June and August. Heterotrophic nanoflagellates (HNF), which accounted for 12% estimated total predation of bacteria at 1 m and 13% at 7m, only exerted a limited predation, mainly by a Monas-type cell. Ciliates, dominated in terms of abundance by Pelagohalteria viridis, accounted for 4% of total predation in the epilimnion (0.00-0.42 x 10⁶ bacteria l^-1 h^-1). In a newly flooded reservoir, metazoan zooplankton appear to be the main consumers of bacteria. Predation of ciliates and HNF by zooplanktonic crustaceans could account for the low contribution of components of the microbial loop to bacterial consumption.      

Feeding of planktonic rotifers on ciliates: a method using natural ciliate assemblages labelled with fluorescent microparticles

A method was developed to allow direct measurements of predationexerted by metazooplankton on ciliates. The method relied onthe use of ciliates labelled with fluorescent microparticles(FMP). Optimal labelling conditions were determined with ciliatesfrom cultures (Tetrahymena pyriformis) and with natural ciliateassemblages sampled in a river. Labelled T. pyriformis wereused as tracer food to determine gut passage time (GPT) andingestion rates of the rotifer Brachionus calyciflorus in thelaboratory. Predation of metazooplankton from the lowland riverMeuse (Belgium) was determined by labelling natural assemblagesof ciliates and using them as tracer food for metazooplankterssampled in the river. Optimal labels of ciliates, i.e. sharpdistribution of FMP in cells, were obtained with short incubations(10 min) and low FMP concentrations (1 x 10⁵ mL^–1). GPTvaried between 30 and 45 min for B. calyciflorus and from 25up to >35 min for rotifers from the river. The ingestionrate of B. calyciflorus fed with T. pyriformis was 3.3 ±0.6 ciliate rot^–1 h^–1, i.e. 1.4 ± 0.3 ngCrot^–1 h^–1. Metazooplankton species for which theingestion of ciliates could be measured were the rotifers Keratellacochlearis, Euchlanis dilatata and Synchaeta spp. Ingestionrates measured ranged from 0.4 to 12.5 ngC rot^–1 h^–1.The method proposed proved to be useful in estimating the predationof microplankton on ciliates in semi- in situ conditions; infurther developments, labelled natural assemblages of ciliatescould be used for in situ incubations with the Haney chamber.