Measuring microzooplankton grazing on planktonic marine bacteria by its impact on bacterial production

Grazing on planktonic bacteria by microzooplankton was estimated by separating bacteria from the larger plankton with 1m pore Nuclepore filtration and measuring changes in bacteria in filtered and unfiltered samples over 24 hours. In the absence of grazers, bacteria increased linearly. The regression coefficient of linear increase was used to estimatein situ bacterial production. When grazers were present, the changes in bacteria concentration usually took the form of a linear decline, and grazing was estimated by subtracting the regression coefficient of the unfiltered sample from that of the 1m filtrate. Results from the Essex estuary-coastal system of northern Massachusetts show grazing and production at rates that indicate a daily turnover of the standing crop of bacteria, with highest values in mid-estuarine waters. Experiments on the size distribution of grazing showed that microzooplankton from 1–3m were responsible for most of the observed decrease in bacteria. It was suggested that the basic pattern of linear increase of the bacteria in the absence of grazing reflects density-dependent limitation by substrate present at the outset of the incubation and is indicative of a population that has been maintained around the mid-point of the logistic growth curve by grazing.     

Phospholipid directed motility of surface-motile bacteria

Myxococcus xanthus is a surface-motile bacterium that has adapted at least one chemosensory system to allow directed movement towards the slowly diffusible lipid phosphatidylethanolamine (PE). The Dif chemosensory pathway is remarkable because it has at least three inputs coupled to outputs that control extracellular matrix (ECM) production and lipid chemotaxis. The methyl-accepting chemotaxis protein, DifA, has two different sensor inputs that have been localized by mutagenesis. The Dif chemosensory pathway employs a novel protein that slows adaptation. Lipid chemotaxis may play important roles in the M. xanthus life cycle where prey-specific and development-specific attractants have been identified. Lipid chemotaxis may also be an important mechanism for locating nutrients by lung pathogens such as Pseudomonas aeruginosa.     

A simplified14C method for grazing measurements on natural planktonic populations

The method used consists of adding highly radioactive material (40 Ci/1 NaH¹⁴CO₃) to sea water with its natural concentrations of zoo- and phytoplankton, incubating this water in the light, separating zoo- from phytoplankton after 1 h or at the most 2 h and measuring the radioactivity of both. Under such conditions, the concentration of the tracer in phytoplankton can be simplified as a linear function of time, and that of the zooplankton as a parabolic function of time. This simplification leads to an overestimation of grazing of at most 2–3%. Comparisons with the Coulter Counter method are given and discussed.     

Propagation of planktonic copepods: production and mortality of eggs

Data on fecundity and egg mortality of neritic copepods were collected in various seasons, areas and under various hydrographical conditions. On a seasonal basis variations in fecundity (F) were related to temperature rather than to the abundance of phytoplankton (P). However, a strong correlation between F and P was evident when water column stability varied horisontally or temporally (i.e. at a tidal front or subsequent to a storm). Estimated specific egg-mortalities were variable and occasionally very severe, up to 9.1 d^–1, implying that down to 10^–4% of the eggs survive to hatching. The implications for phenology and distribution of copepods are discussed.     

Off the hook--how bacteria survive protozoan grazing

Bacterial growth and survival in numerous environments are constrained by the action of bacteria-consuming protozoa. Recent findings suggest that bacterial adaptations against protozoan predation might have a significant role in bacterial persistence and diversification. We argue that selective predation has given rise to diverse routes of bacterial defense, including adaptive mechanisms in bacterial biofilms, and has promoted major transitions in bacterial evolution, such as multicellularity and pathogenesis. We propose that studying predation-driven adaptations will provide an exciting frontier for microbial ecology and evolution at the interface of prokaryotes and eukaryotes.     

Coaggregation between freshwater bacteria within biofilm and planktonic communities

The coaggregation ability of bacteria isolated from a freshwater biofilm was compared to those derived from the coexisting planktonic population. Twenty-nine morphologically distinct bacterial strains were isolated from a 6-month-old biofilm, established in a glass tank under high-shear conditions, and 15 distinct strains were isolated from the associated re-circulating water. All 44 strains were identified to genus or species level by 16S rDNA sequencing. The 29 biofilm strains belonged to 14 genera and 23.4% of all the possible pair-wise combinations coaggregated. The 15 planktonic strains belonged to seven genera and only 5.8% of all the possible pair-wise combinations coaggregated. Therefore, compared to the planktonic population, a greater proportion of the biofilm strains coaggregated. It is proposed that coaggregation influences biofilm formation and species diversity in freshwater under high shear.     

Anaerobic predatory ciliates track seasonal migrations of planktonic photosynthetic bacteria

Abstract Caenomorpha medusula was the dominant anaerobic ciliated protozoon in the anoxic water of a stratified freshwater lake. The population distribution was non-random, often with a peak in the middle of the hypolimnion. Abundance was closely correlated with numbers of the photosynthetic bacterium Thiopedia sp., and the latter was selectively eaten. There was no apparent correlation with the distribution of the total bacterial count, nor with dissolved sulphide. These observations strongly suggest that planktonic anaerobic ciliates seek out specific food sources.     

Growth limitation of planktonic bacteria in a large mesotrophic lake

We studied nutrient-limitation of bacterioplankton growth in Lake Constance, a mesotrophic lake, between February and August in 1992. We amended 1-m filtrates with a single nutrient or nutrient combinations at 5 or 10 m final concentration, and the limiting nutrient or nutrient combination was inferred from the assay in which bacterial growth was most stimulated. The following nutrients were added individually or in combination: glucose, amino acids, peptone, and ammonium as C and N sources, and inorganic phosphate. From January until the beginning of the phytoplankton spring bloom in mid-April, C alone was growth-limiting. During the spring bloom a complex growth-limitation pattern occurred; first P was limiting, then for only 1 week C + N together, and thereafter P + C. During the clear-water phase with very low chlorophyll concentrations, P + C together limited bacterial growth again, interrupted by a period when C + N + P shortage caused a triple limitation. Later in the season, P + C were growth-limiting again. The growth efficiency (bacterial biomass produced/substrates used) on the basis of amino acid and carbohydrate used varied between 17 and 35%. The addition of various C and N sources indicated that the growth efficiency strongly depended on the quality of the substrates and the adaptation of the bacterial assemblages, for example, whether C and N originated from amino acids or glucose and ammonium.     

Use of dissolved carbohydrates by planktonic bacteria in a mesotrophic lake

Dissolved carbohydrates comprise one of the largest pools of labile organic matter readily available for bacterial use in pelagic ecosystems. Despite this fact, very little is known about use of dissolved carbohydrates by planktonic bacteria. We studied use of total dissolved carbohydrates (TDCHO) by planktonic bacteria in mesotrophic Lake Constance, Germany, from April until August 1992. We examined the decrease of TDCHO over time together with the increase of bacterial numbers in 1-m filtered lake water incubated at in situ temperature in the dark. TDCHO analyses were done after hydrolysis by sulfuric acid as free monosaccharides and oxidation by periodate to formaldehyde with 3-methyl-2-benzothiazolinone-hydrazon-hypochloride (MBTH). Preliminary tests showed that hydrolysis by sulfuric acid gave higher yields than hydrolysis by HCl. Our results show that TDCHO are readily used by bacteria and compose substantial fractions of the C requirements for their growth. Concentrations of TDCHO varied between 1.7 and 5.5 m (glucose equivalents), and use rates varied between 0.47 and 3.43 g C liter^–1 h^–1. Highest rates of TDCHO use occurred during the phytoplankton spring bloom, during the clear-water phase in June, and during a phytoplankton bloom in August. Ratios of use of TDCHO/bacterial biomass production varied between 0.17 and 3.05. During the spring bloom, TDCHO and total dissolved amino acids (TDAA) were used in equal amounts. During the clear-water phase at chlorophyll a concentrations <3 g liter^–1, however, bacteria only consumed TDCHO and excreted amino acids. The growth efficiency based on the consumption of TDCHO and TDAA varied between 16 and 21% during the phytoplankton spring bloom and was 35% during the clear-water phase. Correspondence to: M. Simon     

Microbial ecology of planktonic filamentous phototrophic bacteria in holomictic freshwater lakes

The microbial ecology of the filamentous phototrophic bacterium, Chloronema giganteum, has been studied in the water column of three central European lakes (Schlein, Buchen and Vechten). In these lakes an anoxic layer, termed the transition zone, was located between the oxycline and the redoxcline. The migration capacity of Chloronema through this zone appears to be responsible for the natural preponderance of either straight or spiral forms. When the transition zone is less than 1 m thick the straight form is dominant, but when this transition zone is wider than 2 m the spiral form is enriched. The intermediate situations favour both filamentous forms.     

Comparative genomics of DNA fragments from six Antarctic marine planktonic bacteria

Six environmental fosmid clones from Antarctic coastal water bacterioplankton were completely sequenced. The genome fragments harbored small-subunit rRNA genes that were between 85 and 91% similar to those of their nearest cultivated relatives. The six fragments span four phyla, including the Gemmatimonadetes, Proteobacteria (alpha and gamma), Bacteroidetes, and high-G+C gram-positive bacteria. Gene-finding and annotation analyses identified 244 total open reading frames. Amino acid comparisons of 123 and 113 Antarctic bacterial amino acid sequences to mesophilic homologs from G+C-specific and SwissProt/UniProt databases, respectively, revealed widespread adaptation to the cold. The most significant changes in these Antarctic bacterial protein sequences included a reduction in salt-bridge-forming residues such as arginine, glutamic acid, and aspartic acid, reduced proline contents, and a reduction in stabilizing hydrophobic clusters. Stretches of disordered amino acids were significantly longer in the Antarctic sequences than in the mesophilic sequences. These characteristics were not specific to any one phylum, COG role category, or G+C content and imply that underlying genotypic and biochemical adaptations to the cold are inherent to life in the permanently subzero Antarctic waters.     

Flagella and motility behaviour of square bacteria.

Square bacteria are shown to have right-handed helical (RH) flagella. They swim forward by clockwise (CW), and backwards by counterclockwise (CCW) rotation of their flagella. They are propelled by several or single filaments arising at several or single points on the cell surface. When there are several filaments a stable bundle is formed that does not fly apart during the change from clockwise to counterclockwise rotation or vice versa. In addition to the flagella attached to the cells, large amounts of detached flagella aggregated into thick super-flagella, can be observed at all phases of growth.     

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.     

Dynamics of microbial communities on marine snow aggregates: colonization,growth, detachment,and grazing mortality of attached bacteria

We studied the dynamics of microbial communities attached to model aggregates (4-mm-diameter agar spheres) and the component processes of colonization, detachment, growth, and grazing mortality. Agar spheres incubated in raw seawater were rapidly colonized by bacteria, followed by flagellates and ciliates. Colonization can be described as a diffusion process, and encounter volume rates were estimated at about 0.01 and 0.1 cm(3) h(-1) for bacteria and flagellates, respectively. After initial colonization, the abundances of flagellates and ciliates remained approximately constant at 10(3) to 10(4) and approximately 10(2) cells sphere(-1), respectively, whereas bacterial populations increased at a declining rate to >10(7) cells sphere(-1). Attached microorganisms initially detached at high specific rates of approximately 10(-2) min(-1), but the bacteria gradually became irreversibly attached to the spheres. Bacterial growth (0 to 2 day(-1)) was density dependent and declined hyperbolically when cell density exceeded a threshold. Bacterivorous flagellates grazed on the sphere surface at an average saturated rate of 15 bacteria flagellate(-1) h(-1). At low bacterial densities, the flagellate surface clearance rate was approximately 5 x 10(-7) cm(2) min(-1), but it declined hyperbolically with increasing bacterial density. Using the experimentally estimated process rates and integrating the component processes in a simple model reproduces the main features of the observed microbial population dynamics. Differences between observed and predicted population dynamics suggest, however, that other factors, e.g., antagonistic interactions between bacteria, are of importance in shaping marine snow microbial communities.     

Beetle grazing reduces natural infection of Rumex obtusifolius by fungal pathogens

In previously reported laboratory experiments, infection of Rumex obtusifolius by the rust fungus Uromyces rumicis was decreased on leaves which had prior herbivory by the beetle Gastrophysa viridula . In this paper we investigate whether this interaction is robust for natural infection by a variety of fungi in field experiments carried out in spring and autumn with plants given different levels of nitrogen fertilization. Grazing by G. viridula led to a decrease in lesion density of Ramularia rubella and Venturia rumicis in the spring and V. rumicis and U. rumicis in the autumn experiment. For V. rumicis and U. rumicis significant reductions in lesion density occurred on the undamaged leaves of damaged plants, compared with similar leaves on undamaged plants, suggesting systemic induced resistance. This induced resistance was usually independent of the amount of nitrogen fertilization, although the inhibitory effect of grazing on R. rubella in the spring and V. rumicis in the autumn experiment was enhanced by increasing nitrogen fertilization and was inhibited by increasing nitrogen fertilization for V. rumicis in the spring. In both experiments, the lesion density of V. rumicis was greater on leaves on which R. rubella was also present, and the presence of U. rumicis in the autumn experiment was linked to a similar but greater effect on V. rumicis lesion density. We found no evidence of induced resistance by fungi against fungi in these experiments. We highlight the complex interactions between inhibitory and facilitatory processes acting on leaf fungal infection. These results are compared with the proposed molecular mechanisms of induced resistance(s) and we consider the benefits of closer integration between molecular and ecological investigations of induced resistances that occur in the field.     

Flagellate grazing on bacteria in a small dystrophic lake

Fluorescent beads were used to determine the grazing on bacteria by heterotrophic and mixotrophic flagellates in a highly humic (water colour 300–600 mg Pt l^–1) small lake. In summer phagotrophic flagellates constituted about three quarters of the numbers of phytoplankton (including heterotrophic or mixotrophic flagellates) in the uppermost epilimnion. Due to their small size their respective contribution to the biomass was about one quarter. The most important phagotrophic species were Ochromonas sp., and Chromulina spp. which ingested 75–203% of their body carbon per day from bacteria.In view of the abundance and biomass of phagotrophic and mixotrophic flagellates and their very high growth potential, they clearly play a significant role in the food chains of this lake. In addition to providing energy, bacteriovory also represents an important supply of inorganic and organic nutrients under nutrient limiting conditions.     

The relationship between planktonic algae and bacteria in a small lake

The seasonal changes and vertical distribution of the aerobic and anaerobic bacteria in a small edaphically eutrophic lake which exhibited thermal and chemical stratification are described. There was some correspondence between the phytoplankton and particularly the aerobic bacteria but this was not consistent. Increases in the numbers of anaerobic bacteria coincided with the low dissolved oxygen concentrations in the hypolimnion when algal populations were first senescent and then increasing actively in size.     

Morphological and compositional changes in a planktonic bacterial community in response to enhanced protozoan grazing

We analyzed changes in bacterioplankton morphology and composition during enhanced protozoan grazing by image analysis and fluorescent in situ hybridization with group-specific rRNA-targeted oligonucleotide probes. Enclosure experiments were conducted in a small, fishless freshwater pond which was dominated by the cladoceran Daphnia magna. The removal of metazooplankton enhanced protozoan grazing pressure and triggered a microbial succession from fast-growing small bacteria to larger grazing-resistant morphotypes. These were mainly different types of filamentous bacteria which correlated in biomass with the population development of heterotrophic nanoflagellates (HNF). Small bacterial rods and cocci, which showed increased proportion after removal of Daphnia and doubling times of 6 to 11 h, belonged nearly exclusively to the beta subdivision of the class Proteobacteria and the Cytophaga-Flavobacterium cluster. The majority of this newly produced bacterial biomass was rapidly consumed by HNF. In contrast, the proportion of bacteria belonging to the gamma and alpha subdivisions of the Proteobacteria increased throughout the experiment. The alpha subdivision consisted mainly of rods that were 3 to 6 microm in length, which probably exceeded the size range of bacteria edible by protozoa. Initially, these organisms accounted for less than 1% of total bacteria, but after 72 h they became the predominant group of the bacterial assemblage. Other types of grazing-resistant, filamentous bacteria were also found within the beta subdivision of Proteobacteria and the Cytophaga-Flavobacterium cluster. We conclude that the predation regimen is a major structuring force for the bacterial community composition in this system. Protozoan grazing resulted in shifts of the morphological as well as the taxonomic composition of the bacterial assemblage. Grazing-resistant filamentous bacteria can develop within different phylogenetic groups of bacteria, and formerly underepresented taxa might become a dominant group when protozoan predation is the major selective pressure.