Patchiness in the Distribution of Planktonic Heterotrophic Bacteria in Lakes

By using conventional dilution plating procedures, the heterotrophic planktonic bacteria in Lake Washington were found to be distributed in small-scale (<1.0 ml) and large-scale (>1 m in horizontal distance) patches. Recommendations are made on sampling and enumeration procedures to minimize the effect of patchiness on obtaining an accurate estimate of bacterial numbers.     

Distribution of Heterotrophic Bacteria in Lake Shira

A study of the horizontal and vertical distribution of heterotrophic bacteria in brackish Lake Shira in summer periods showed that mesophilic bacteria dominated in all areas of the lake, whereas psychrotolerant bacteria dominated in the metalimnion and hypolimnion of its central part. Nonhalophilic bacteria were mostly mesophilic and dominated in coastal waters. Most psychrotolerant bacteria were able to grow in the presence of 5–10% NaCl. Heterotrophic bacteria isolated in different regions of the lake were identified to a generic level. The isolates were classified into autochthonous and allochthonous microorganisms on the bases of their distribution pattern in the lake water, halotolerance, and ability to grow at low temperatures.     

Massive Regime Shifts and High Activity of Heterotrophic Bacteria in an Ice-Covered Lake

In winter 2009/10, a sudden under-ice bloom of heterotrophic bacteria occurred in the seasonally ice-covered, temperate, deep, oligotrophic Lake Stechlin (Germany). Extraordinarily high bacterial abundance and biomass were fueled by the breakdown of a massive bloom of Aphanizomenon flos-aquae after ice formation. A reduction in light resulting from snow coverage exerted a pronounced physiological stress on the cyanobacteria. Consequently, these were rapidly colonized, leading to a sudden proliferation of attached and subsequently of free-living heterotrophic bacteria. Total bacterial protein production reached 201 µg C L⁻¹ d⁻¹, ca. five times higher than spring-peak values that year. Fluorescence in situ hybridization and denaturing gradient gel electrophoresis at high temporal resolution showed pronounced changes in bacterial community structure coinciding with changes in the physiology of the cyanobacteria. Pyrosequencing of 16S rRNA genes revealed that during breakdown of the cyanobacterial population, the diversity of attached and free-living bacterial communities were reduced to a few dominant families. Some of these were not detectable during the early stages of the cyanobacterial bloom indicating that only specific, well adapted bacterial communities can colonize senescent cyanobacteria. Our study suggests that in winter, unlike commonly postulated, carbon rather than temperature is the limiting factor for bacterial growth. Frequent phytoplankton blooms in ice-covered systems highlight the need for year-round studies of aquatic ecosystems including the winter season to correctly understand element and energy cycling through aquatic food webs, particularly the microbial loop. On a global scale, such knowledge is required to determine climate change induced alterations in carbon budgets in polar and temperate aquatic systems.     

Heterotrophic Activity of Deep-Sea Sediment Bacteria

Sediment samples, containing mixed microbial populations that were decompressed during retrieval from 7,750 and 8,130 m in the Puerto Rican Trench, were recompressed and incubated at the approximate in situ temperature (3 C) and pressure (775 or 815 atm) in the presence of 14C-labeled amino acids. Heterotrophic activity (total uptake, CO2 respiration, and cellular assimilation) and cellular-associated "pool" concentrations were measured. Compared with atmospheric controls held at 3 C, the total uptake at elevated pressure at 3 C was reduced, on an average, 55 times, CO2 respiration was reduced 45 times, and cellular assimilation was reduced 69 times. Rate of total uptake at elevated pressure was found to range from 4.0 X 10(-11) mug/cell per h for leucine to 2.61 X 10(-10) mug/cell per h for an amino acid mixture. Also, the percentage of total uptake at elevated pressures, respired as CO2, increased at the expense of cellular assimilation (ca. 22% increase). Two cellular-associated amino acid pools were detected, a large, loosely bound, outer pool and a small, tightly bound internal pool. The loosely bound outer pool was removed by a change in the pH of the incubation medium. Even though heterotrophic uptake and the outer, cellular-associated pool were markedly reduced at an elevated pressure, the percentage of total uptake calculated for the unincorporated, tightly bound, intracellular pool was 2 to 19 times that obtained for cultures held at 1 atm. The results were interpreted as indicating that bacterial metabolism and biosynthesis in the deep sea are markedly reduced, with a greater proportion of metabolic activity devoted to cellular maintenance.     

Permanent Presence of Grazing-Resistant Bacteria in a Hypertrophic Lake

The size structure of planktonic bacteria from a hypertrophic lake was investigated at 5- to 15-day intervals by means of a semiautomatic image analysis system during 1 year. Characteristic of this bacterial assemblage was the permanent presence of large filamentous bacteria and small cocci with cell sizes of <0.01 (mu)m(sup3). These filamentous bacteria, sometimes longer than 200 (mu)m and with cell volumes of up to 276 (mu)m(sup3), are larger than nanoflagellates (<20 (mu)m) and, even, metazoans living in the lake. Although they account for only 4 to 16% of bacterial abundance, their contribution to total bacterial biovolume was between 45 and 86%. An analysis of the food web structure indicates that this particular bacterial size structure may be the consequence of a strong bacterivory pressure by nanoflagellates and the absence of other larger bacterivores. The persistence of bacterial forms resistant to grazing has important consequences for the carbon flow within the microbial food web.     

Vertical and Seasonal Dynamics of Planktonic Ciliates in a Strongly Stratified Hypertrophic Lake

Seasonal population dynamics and the vertical distribution of planktonic ciliates in a hypertrophic and strongly stratified temperate lake were studied from April to October in 2000 and from April to June in 2001. In the epi- and metalimnion the ciliate abundance peaked in spring and late summer, reaching maximum values in the metalimnion (86 cells ml⁻¹) on 7th August 2000. In the epilimnion, the highest biomass content (414 μg C l⁻¹) was observed on 8th May 2000. In the hypolimnion only a late summer peak occurred and the ciliate numbers were always lower than in the epi- and metalimnion. Five groups dominated the community of ciliates: Oligotrichida, Gymnostomatea, Prostomatida, Hymenostomata and Peritrichia, and the community composition varied greatly with depth. In the epilimnion the ciliate numbers were dominated by oligotrichs but small algivorous prostomatids, peritrichs and gymnostomes were also numerous. In the metalimnion these groups were gradually replaced by scuticociliates and mixotrophic Coleps spp. In the hypolimnion scuticociliates and species known as benthic migrants dominated. In the epilimnion and upper metalimnion in spring large herbivores and in summer small bacterivores were more numerous.     

Impact of Storms on Heterotrophic Activity of Epilimnetic Bacteria in a Southwestern Reservoir

The impact of storm conditions on the heterotrophic activity of planktonic bacteria in a southwestern reservoir was investigated. Storm events were considered as rainfall in excess of 2.5 cm in a 24-h period before sampling. Storm conditions stimulated heterotrophic activities and resulted in increased uptake rates and decreased turnover times of glutamate and acetate. Uptake rates were 45 to 75% faster immediately after storm conditions than they were during calm conditions. Activity levels appeared to return to prestorm levels within 48 h. Bacterial cell numbers did not change substantially during storm events. Cell-specific activity indicated that increases in heterotrophic activity were the result of increased activity of individual cells. Light penetration, levels of particulate organic carbon, K_t + S_n values, and population levels of attached bacteria suggest that immediate sediment loading of the reservoir or increased substrate levels could not account for abrupt increases in heterotrophic activities.     

Bacterioplankton Abundance and Activity in a Small Hypertrophic Stratified Lake

Bacterioplankton abundance and production were followed during one decade (1991–2001) in the hypertrophic and steeply stratified small Lake Verevi (Estonia). The lake is generally dimictic. However, a partly meromictic status could be formed in specific meteorological conditions as occurred in springs of 2000 and 2001. The abundance of bacteria in Lake Verevi is highly variable (0.70 to 22 × 10⁶ cells ml⁻¹) and generally the highest in anoxic hypolimnetic water. In 2000–2001, the bacterial abundance in the hypolimnion increased probably due to meromixis. During a productive season, heterotrophic bacteria were able to consume about 10–40% of primary production in the epilimnion. Our study showed that bacterioplankton in the epilimnion was top-down controlled by predators, while in metalimnion bacteria were dependent on energy and carbon sources (bottom-up regulated). Below the thermocline hypolimnetic bacteria mineralized organic matter what led to the depletion of oxygen and created anoxic hypolimnion where rich mineral nutrient and sulphide concentrations coexisted with high bacterial numbers.     

Diversity of Heterotrophic Nitrogen Fixation Genes in a Marine Cyanobacterial Mat

The diversity of nitrogenase genes in a marine cyanobacterial mat was investigated through amplification of a fragment of nifH, which encodes the Fe protein of the nitrogenase complex. The amplified nifH products were characterized by DNA sequencing and were compared with the sequences of nitrogenase genes from cultivated organisms. Phylogenetic analysis showed that similar organisms clustered together, with the exception that anaerobic bacteria clustered together, even though they represented firmicutes, (delta)-proteobacteria, and (gamma)-proteobacteria. Mat nifH sequences were most closely related to those of the anaerobes, with a few being most closely related to the cluster of (gamma)-proteobacteria containing Klebsiella and Azotobacter species. No cyanobacterial nifH sequences were found from the mat collected in November when Microcoleus chthonoplastes was the dominant cyanobacterium, but sequences closely related to the cyanobacterium Lyngbya lagerheimeii were found during summer, when a Lyngbya strain was dominant. The results indicate that there is a high diversity of heterotrophic nitrogen-fixing organisms in marine cyanobacterial mats.     

Response of Heterotrophic Planktonic Bacteria to the Zebra Mussel Invasion of the Tidal Freshwater Hudson River

Abstract Invasions of aquatic ecosystems by exotic bivalves are known to cause dramatic changes in phytoplankton and some other groups, but their effect on the microbial component is unknown. The invasion of the tidal freshwater Hudson River by the exotic zebra mussel (Dreissena polymorpha) has caused large changes in several components of the Hudson's food web. Planktonic bacteria in the tidal freshwater Hudson are a major part of the food web, and mediate important processes in the carbon budget. We used a long-term data set, spanning four years prior to the zebra mussel (ZM) invasion and four years post-invasion, to describe ZM effects on planktonic bacteria. Small and meso-scale experiments were conducted to specifically examine direct consumption of bacteria by ZM, as well as effects on protozoans. Bacterial abundances in the Hudson have increased roughly 2× since the ZM arrived, making it clear that direct consumption by Dreissena is a minor process. Experiments show that ZM do not remove bacteria from Hudson River water, but are very effective at clearing flagellated protozoans, the major predator of bacteria. The observed changes in bacterial abundance have not been accompanied by equally large changes in bacterial productivity, suggesting growth is primarily limited by carbon supply. Bacterial production has not declined despite a dramatic decline of phytoplankton, confirming previous suggestions that bacteria and phytoplankton are not strongly linked in the Hudson. As a result of the increase in bacterial abundance and removal of phytoplankton, the absolute and relative contributions of bacterial carbon to living particulate organic carbon (POC) standing stocks have increased dramatically. The maintenance of the bacterial component of the Hudson River's food web may be one mechanism whereby consumers are ``insulated' from effects of zebra mussel consumption of phytoplankton carbon. Received: 24 October 1997; Accepted: 9 February 1998     

Purple Sulfur Bacteria Control the Growth of Aerobic Heterotrophic Bacterioplankton in a Meromictic Salt Lake

In meromictic Mahoney Lake, British Columbia, Canada, the heterotrophic bacterial production in the mixolimnion exceeded concomitant primary production by a factor of 7. Bacterial growth rates were correlated neither to primary production nor to the amount of chlorophyll a. Both results indicate an uncoupling of bacteria and phytoplankton. In the chemocline of the lake, an extremely dense population of the purple sulfur bacterium Amoebobacter purpureus is present year round. We investigated whether anoxygenic phototrophs are significant for the growth of aerobic bacterioplankton in the overlaying water. Bacterial growth rates in the mixolimnion were limited by inorganic phosphorus or nitrogen most of the time, and the biomass of heterotrophic bacteria did not increase until, in autumn, 86% of the cells of A. purpureus appeared in the mixolimnion because of their reduced buoyant density. The increase in heterotrophic bacterial biomass, soluble phosphorus concentrations below the detection limit, and an extraordinarily high activity of alkaline phosphatase in the mixolimnion indicate a rapid liberation of organically bound phosphorus from A. purpureus cells accompanied by a simultaneous incorporation into heterotrophic bacterioplankton. High concentrations of allochthonously derived dissolved organic carbon (mean, 60 mg of C(middot)liter(sup-1)) were measured in the lake water. In Mahoney Lake, liberation of phosphorus from upwelling purple sulfur bacteria and degradation of allochthonous dissolved organic carbon as an additional carbon source render heterotrophic bacterial production largely independent of the photosynthesis of phytoplankton. A recycling of inorganic nutrients via phototrophic bacteria also appears to be relevant in other lakes with anoxic bottom waters.     

Detection and Quantification with 16S rRNA Probes of Planktonic Methylotrophic Bacteria in a Floodplain Lake

Abstract Two approaches employing 16S rRNA oligonucleotide probes, in situ hybridization combined with ³³P-autoradiography and ³²P-labeled slot-blot hybridizations on nitrocellulose filters, were used to enumerate methylotrophic bacteria in the water column of Ryans 1 Billabong, a small floodplain lake in northeastern Victoria, Australia. Methylotrophic bacterioplankton numbered 0.6–1.2 × 10⁹ cells liter⁻¹ in the winter of 1994, and 0.8–5.5 × 10⁹ cells liter⁻¹ in the summer of 1994–95. This was equivalent to 10–46% of total bacterioplankton cell counts, determined via epifluorescence microscopy. Methylotrophic bacteria were not detected in the water column of the nearby Kiewa River, and a set of laboratory controls indicated that the high abundance of methylotrophs in the billabong samples was not a methodological artifact. There was no change, with water depth, in total bacterioplankton or methylotroph abundance in winter, a result consistent with the water column being well mixed at this time of year (dissolved O₂ concentrations 5–7 mg liter⁻¹; dissolved methane concentrations <60 μg liter⁻¹, or <5% methane saturation, at all depths). In summer the billabong became diurnally stratified (dissolved O₂ concentrations <2 mg liter⁻¹ at water depths of >45 cm; dissolved methane concentrations <100 μg liter⁻¹ at the surface, but >500 μg liter⁻¹ near the sediments) and there was a correspondingly marked increase in the abundance of total bacterioplankton and methylotrophs with depth. In situ hybridizations and slot-blot hybridizations both indicated that type II methylotrophs (detected with a probe specific for the 9-α subgroup of Proteobacteria) were markedly less abundant than were type I and X methylotrophs (detected with a probe specific for the 10-γ subgroup of Proteobacteria). Received: 12 March 1996; Accepted: 2 October 1996     

Influence of Three Contrasting Detrital Carbon Sources on Planktonic Bacterial Metabolism in a Mesotrophic Lake

Abstract Lakes receive organic carbon from a diversity of sources which vary in their contribution to planktonic microbial food webs. We conducted a mesocosm study to test the effects of three different detrital carbon sources (algae, aquatic macrophytes, terrestrial leaves) on several measures of microbial metabolism in a small meso-eutrophic lake (DOC ≈ 5 mg/L). Small DOC additions (ΔC < 1 mg/L) affected bacterial numbers, growth, and pathways of carbon acquisition. Macrophyte and leaf detritus significantly increased TDP and color, but bacterial densities initially (+12 h) were unaffected. After 168 h, densities in systems amended with terrestrial detritus were 60% less than in controls, while production rates in mesocosms with macrophyte detritus were 4-fold greater. Detritus treatments resulted in greater per-cell production rates either through stable cell numbers and greater growth rates (macrophyte-C) or lower densities with stable production rates (terrestrial-C). After only 12 h, rates of leucine aminopeptidase (LAPase) activity were 2.5× greater in macrophyte-C systems than in controls, but LAPase and β-N-acetylglucosamindase activities in systems amended with terrestrial-C were only 50% of rates in controls. After 168 h, β-xylosidase rates were significantly greater in communities with terrestrial and phytoplankton detritus. Microbial utilization of >20% of 102 carbon sources tested were affected by at least one detritus addition. Macrophyte-C had positive (6% of substrates) and negative (14%) effects on substrate use; terrestrial detritus had mainly positive effects. An ordination based on carbon-use profiles (+12 h) revealed a cluster of macrophyte-amended communities with greater use of psicose, lactulose, and succinamic acid; controls and algal-detritus systems were more effective in metabolizing two common sugars and cellobiose. After 168 h, communities receiving terrestrial detritus were most tightly clustered, exhibiting greater use of raffinose, pyroglutamic acid, and sebacic acid. Results suggest that pelagic bacterial communities respond to changes in organic carbon source rapidly and by different routes, including shifts in per-cell production rates and variations in degradation of a variety of compounds comprising the DOC pool. Received: 5 June 1998; Accepted: 24 August 1998     

Extracellular Phytoplankton Production and Its Importance for Heterotrophic Activity of Bacteria

Biology Bulletin - The ecological role of the extracellular production of phytoplankton is discussed. The experiments based on a radiocarbon method combined with differential filtration of water...     

Correlation of Direct Viable Counts with Heterotrophic Activity for Marine Bacteria

Viable-bacteria counts, heterotrophic activity, and substrate responsiveness of viable bacteria have been used to measure microbial activity. However, the relationship between these parameters is not clear. Thus, the direct viable count (DVC) method was used to analyze seawater samples collected from several different geographical locations. Samples collected from offshore waters of the South China Sea and western Pacific Ocean yielded DVC that indicated the presence of surface and subsurface peaks of viable, substrate-responsive bacteria which could be correlated with turnover rates of amino acids obtained by using uniformly ¹⁴C-labeled amino acids. DVC were always less than total viable counts (acridine orange direct counts), and the DVC subsurface peak occurred close to and within the chlorophyll a zone, suggesting algal-bacterial interactions within the layer. For comparison with the open-ocean samples, selected substrates were used to determine the response of viable bacteria present in seawater samples collected near an ocean outfall of the Barceloneta Regional Waste Treatment Plant, Barceloneta, Puerto Rico. The number of specific substrate-responsive bacteria at the outfall stations varied depending on the substrate used and the sampling location. Changes in the population size or physiological condition of the bacteria were detected and found to be associated with the presence of pharmaceutical waste.     

Production Rate of Planktonic Bacteria in the North Basin of Lake Biwa, Japan

Vertical and seasonal variations in the cell number and production rate of planktonic bacteria were investigated at a pelagic site (water depth, ca. 72 m) of the north basin of Lake Biwa during April to October 1986. The [methyl-³H]thymidine uptake rate into a cold trichloroacetic acid-insoluble fraction and the frequency of dividing cells (FDCs) were measured for each sample as indices of the bacterial production rate. The seasonal data of bacterial number, thymidine uptake rate, and bacterial growth rate based on the FDCs were correlated with one another (rank correlation analysis, P < 0.05). These bacterial variables were not correlated positively with the chlorophyll a concentration. Vertically, the maxima of both bacterial number and the thymidine uptake rate were found in the euphotic zone. The direct counting of bacteria and the measurements of thymidine uptake rate combined with the size-fractionation method revealed that more than 90% of the bacterial biomass and production rate were attributed to unattached bacteria throughout the investigation period. The carbon flux estimates of bacterial production were less certain due to the variability of the conversion factor for the thymidine uptake method and that of the calibration for the FDC method, but even when the conservative range of bacterial net production rate was used (5 to 60 μg of carbon per liter per day), it can be suggested that bacterial net production in the investigated area was a significant fraction (ca. 30%) of the level of the primary production rate in the same water basin.     

Measurement and Significance of Specific Activity in the Heterotrophic Bacteria of Natural Waters

It is now possible to obtain accurate total counts of the bacteria of natural waters with the use of acridine orange staining and epifluorescence microscopy. This approach can be coupled to highly sensitive measurements of heterotrophic activity using radioisotopes. To accomplish this, three variations of a “specific activity index” are suggested, based on different approaches to measuring heterotrophic activity with radiolabeled organic solutes. The denominator of each index is the direct count of bacteria from a given natural sample. Three numerators are presented, each of which has been shown to vary directly with heterotrophic bacterial activity: V_max, turnover rate, and direct uptake (at high substrate concentrations). Each approach is illustrated with data from estuarine and coastal waters of northeastern Massachusetts. The data show major differences in specific activity that accompany such habitat differences as distances within or offshore from an estuary and vertical location in the water column. These and other data suggest that specific activity is a valid indicator of the physiological state and metabolic role of the bacteria. Some evidence is presented in support of the hypothesis that the natural bacteria are adapted to conditions of nutrient starvation by becoming “dormant,” existing for an unknown period of time in a reversible physiological state that reflects the availability of organic nutrients.     

Temporal and Vertical Difference in Factors Limiting Growth Rate of Heterotrophic Bacteria in Lake Biwa

Abstract Dilution bioassays were performed to examine the seasonal and vertical difference in the relative importance of factors limiting growth of heterotrophic bacteria in Lake Biwa. The lake water diluted by 0.2 μm lake filtrate (1:6.6) was enriched either with glucose (C), inorganic phosphorus (P), ammonium nitrogen (N), amino acids (AA), or a combination of these, and incubated for 2 days at the depths where lake water was collected (2.5, 20 and 30 m depths). Experiments showed that at 2.5 m, P was the most deficient resource for bacterial growth, but the magnitude of P limitation depended on water temperature. Among others, amino acids showed a slight but significant stimulation of bacterial growth rates during the fall. At 20 and 30 m, however, growth stimulation by resource addition was rarely detected. Vertically reciprocal translocation experiments revealed that the growth rate was limited by low temperature rather than resource supply at the greater depths. The results support a simple view that bacterial growth rate is basically regulated by water temperature, but high growth rate is not realized in summer because of resource depletion. The present study suggests that both temperature and P supply play a crucial role in biogeochemical cycling of organic matter in Lake Biwa through the bacterial growth rate. Received: 10 March 1999; Accepted: 14 May 1999     

The Ecological Roles of Heterotrophic Dinoflagellates in Marine Planktonic Community1

Heterotrophic dinoflagellates are ubiquitous and often abundant protists in marine environments. Recently, several novel predator-prey relationships between heterotrophic dinoflagellates and other planktonic organisms have been discovered and shown to have diverse ecological roles. Heterotrophic dinoflagellates are predators on a wide array of prey items, including phytoplankton, copepod eggs, and early naupliar stages. They are in turn important prey for some metazoa. Some heterotrophic dinoflagellates are predators of and simultaneously prey for other dinoflagellates. These newly discovered predator-prey relationships may influence our conventional view of energy flow and carbon cycling in the marine planktonic community.     

Survival and Activity of Bacteria in a Deep,Aged Lake Sediment (Lake Constance)

Abstract Viable counts and potential activities of different bacteria were determined as a function of depth in the deep profundal sediment of Lake Constance, Germany. The sediment layer at the bottom of the lake had a total depth of about 7 m and was deposited in the time after the last ice age, i.e., over the past 13,000 years. The high clay content of the sediment prevents seepage. Below 25 cm all of the viable heterotrophic bacteria were present as heat-resistant spores. Numbers of viable spores of both aerobic and anaerobic heterotrophic bacteria decreased exponentially with sediment depth and were below the detection limit (5–55 cells ml⁻¹) at 4–6 m, i.e., in about 8,900-year-old sediment. Absence of viable heterotrophic bacteria in deeper sediment layers demonstrated that aseptic sampling conditions were achieved. The decrease of viable spores with depth may be interpreted as time-dependent death of spores resulting in a death rate of about 0.0013–0.0025 year⁻¹. Viable units of specific metabolic groups of bacteria were detected only in the upper sediment layers (0–50 cm). Nitrifying bacteria could not be detected below 30 cm. Methane-oxidizing bacteria were present in the sediment down to >30 cm, but were in a dormant state. Nitrate reduction activity decreased by a factor of 6 within the upper 25 cm of the sediment, but was still detected at 50 cm. Sulfate reduction, on the other hand, could not be detected at depths of 20 cm and below. By contrast, methanogenesis and methanogenic bacteria could be detected down to 50 cm. These observations indicate that bacteria eventually become nonviable in aged sediments. Received: 5 March 1996; Accepted: 12 March 1996