Biodiversity on the brink: an assessment of conservation strategies for North American freshwater mussels

Fish and zooplankton populations of nine Ethiopian freshwater lakes were quantitatively sampled along a North–South gradient. Differences in altitude and latitude resulted in a temperature gradient from North to South. We tested three hypotheses: (1) the degree of zooplanktivory decreases with water temperature, i.e. from North to South; (2) the degree of zooplanktivory increases with the abundance of large-bodied zooplankton; and (3) the pattern of zooplanktivory in eutrophic Ethiopian water bodies differs from other tropical and temperate water bodies. Proportions of zooplanktivory in the fish communities did not show a geographical trend, but mainly depended on fish species, zooplankton density and the availability of large-bodied cladocerans. The degree of zooplanktivory in eutrophic Ethiopian water bodies differs from other eutrophic water bodies, both temperate and tropical. In Ethiopia, the degree of zooplanktivory can be both low and high, in contrast with other tropical water bodies where zooplanktivory is generally low and with temperate eutrophic water bodies where it is generally high. As a result, predation pressure on zooplankton by fish varies dramatically amongst Ethiopian water bodies.     

Pronounced seasonal dynamics of freshwater chitinase genes and chitin processing

Seasonal variation in activity of enzymes involved in polymer degradation, including chitinases, has been observed previously in freshwater environments. However, it is not known whether the seasonal dynamics are due to shifts in the activity of bacteria already present, or shifts in community structure towards emergence or disappearance of chitinolytic organisms. We traced seasonal shifts in the chitinase gene assemblage in a temperate lake and linked these communities to variation in chitinase activity. Chitinase genes from 20 samples collected over a full yearly cycle were characterized by pyrosequencing. Pronounced temporal shifts in composition of the chitinase gene pool (beta diversity) occurred along with distinct shifts in richness (alpha diversity) as well as chitin processing. Changes in the chitinase gene pool correlated mainly with temperature, abundance of crustacean zooplankton and phytoplankton blooms. Also changes in the physical structure of the lake, e.g. stratification and mixing were associated with changes in the chitinolytic community, while differences were minor between surface and suboxic hypolimnetic water. The lake characteristics influencing the chitinolytic community are all linked to changes in organic particles and we suggest that seasonal changes in particle quality and availability foster microbial communities adapted to efficiently degrade them.     

Community structure and diel migration of zooplankton in shallow brackish lakes: role of salinity and predators

Diel horizontal migration (DHM), where zooplankton moves towards macrophytes during daytime to avoid planktivorous fish, has been reported as a common migration pattern of zooplankton in shallow temperate freshwater lakes. However, in shallow eutrophic brackish lakes, macrophytes seem not to have the same refuge effect, as these lakes may remain turbid even at relatively high macrophyte abundances. To investigate the extent to which macrophytes serve as a refuge for zooplankton at different salinities, we introduced artificial plants mimicking submerged macrophytes in the littoral zone of four shallow lakes, with salinities ranging from almost freshwater (0.3) to oligohaline waters (3.8). Furthermore, we examined the effects of different salinities on the community structure. Diel samples of zooplankton were taken from artificial plants, from areas where macrophytes had been removed (intermediate areas) and, in two of the lakes, also in open water. Fish and macroinvertebrates were sampled amongst the artificial plants and in intermediate areas to investigate their influence on zooplankton migration. Our results indicated that diel vertical migration (DVM) was the most frequent migration pattern of zooplankton groups, suggesting that submerged macrophytes were a poor refuge against predation at all salinities under study. Presumably, this pattern was the result of the relatively high densities of small planktivorous fish and macroinvertebrate predators within the submerged plants. In addition, we found major differences in the composition of zooplankton, fish and macroinvertebrate communities at the different salinities and species richness and diversity of zooplankton decreased with increasing salinity. At low salinities both planktonic/free-swimming and benthic/plant-associated cladocerans occurred, whilst only benthic ones occurred at the highest salinity. The low zooplankton biomass and overall smaller-bodied zooplankton specimens may result in a lower grazing capacity on phytoplankton, and enhance the turbid state in nutrient rich shallow brackish lakes.     

Mining of unexplored habitats for novel chitinases—chiA as a helper gene proxy in metagenomics

The main objective of this study was to assess the abundance and diversity of chitin-degrading microbial communities in ten terrestrial and aquatic habitats in order to provide guidance to the subsequent exploration of such environments for novel chitinolytic enzymes. A combined protocol which encompassed (1) classical overall enzymatic assays, (2) chiA gene abundance measurement by qPCR, (3) chiA gene pyrosequencing, and (4) chiA gene-based PCR-DGGE was used. The chiA gene pyrosequencing is unprecedented, as it is the first massive parallel sequencing of this gene. The data obtained showed the existence across habitats of core bacterial communities responsible for chitin assimilation irrespective of ecosystem origin. Conversely, there were habitat-specific differences. In addition, a suite of sequences were obtained that are as yet unregistered in the chitinase database. In terms of chiA gene abundance and diversity, typical low-abundance/diversity versus high-abundance/diversity habitats was distinguished. From the combined data, we selected chitin-amended agricultural soil, the rhizosphere of the Arctic plant Oxyria digyna and the freshwater sponge Ephydatia fluviatilis as the most promising habitats for subsequent bioexploration. Thus, the screening strategy used is proposed as a guide for further metagenomics-based exploration of the selected habitats.     

Quantitative development of zooplankton in waterbodies and watercourses of the Great Lakes Depression (Mongolia)

The indices of the zooplankton of various waterbodies and watercourses of the Great Lakes Depression (Mongolia) are analyzed. The maximum quantity of zooplankton is registered in freshwater bodies. Irrespective of the waterbody type, the greatest number and biomass of zooplankton are recorded in the littoral zone, where communities are characterized by a high abundance of rotifers, including the indicator species of eutrophic waters. This indicates a high degree of organic load from the catchment area. In comparison with the data obtained by the end of 1970s, the zooplankton biomass decreased with a concomitant increases in the shares of rotifers and cladocerans. These changes are most pronounced in the freshwater ecosystem. The quantitative development of the zooplankton of watercourses depends on the character of the river head, the location of sampling stations along the watercourse’s channel, and the water current velocity.     

Cloning, expression, and characterization of a chitinase from the chitinolytic bacterium Aeromonas hydrophila strain SUWA-9

The chitinolytic bacterium Aeromonas hydrophila strain SUWA-9, which was isolated from freshwater in Lake Suwa (Nagano Prefecture, Japan), produced several kinds of chitin-degrading enzymes. A gene coding for an endo-type chitinase (chiA) was isolated from SUWA-9. The chiA ORF encodes a polypeptide of 865 amino acid residues with a molecular mass of 91.6 kDa. The deduced amino acid sequence showed high similarity to those of bacterial chitinases classified into family 18 of glycosyl hydrolases. chiA was expressed in Escherichia coli and the recombinant chitinase (ChiA) was purified and examined. The enzyme hydrolyzed N-acetylchitooligomers from trimer to pentamer and produced monomer and dimer as a final product. It also reacted toward colloidal chitin and chitosan with a low degree of deacetylation. When cells of SUWA-9 were grown in the presence of colloidal chitin, a 60 kDa-truncated form of ChiA that had lost the C-terminal chitin-binding domain was secreted.     

Changes and drivers of zooplankton diversity patterns in the middle reach of Yangtze River floodplain lakes,China

Anthropogenic habitat alteration interferes the natural aquatic habitats and the system''s hydrodynamics in the Yangtze River floodplain lakes, resulting in a serious decline in freshwater biodiversity. Zooplankton communities possess major position in freshwater ecosystems, which play essential parts in maintaining biological balance of freshwater habitats. Knowledge of processes and mechanisms for affecting variations in abundance, biomass, and diversity of zooplankton is important for maintaining biological balance of freshwater ecosystems. Here, we analyzed that the temporal and spatial changes in the structure of zooplankton community and their temporal and spatial variations respond to changes in environmental factors in the middle reach of Yangtze River floodplain lakes. The results showed that zooplankton samples were classified into 128 species, and Rotifera was the most common taxa. Significant seasonal differences were found among the abundance and diversity of zooplankton. Similarly, we also found significant seasonal differences among the biomass of zooplankton functional groups. The spatial turnover component was the main contributor to the β diversity pattern, which indicated that study areas should establish habitat restoration areas to restore regional biodiversity. The NMDS plot showed that the structure of zooplankton community exhibited significant seasonal changes, where the community structure was correlated with pH, water temperature, water depth, salinity, total nitrogen, chlorophyll‐a, and total phosphorus based on RDA. This study highlights that it is very important to ensure the floodplain ecosystem''s original state of functionality for maintaining the regional diversity of the ecosystem as a whole.     

Differentiation of chitinase-active and non-chitinase-active subpopulations of a marine bacterium during chitin degradation

The ability of marine bacteria to adhere to detrital particulate organic matter and rapidly switch on metabolic genes in an effort to reproduce is an important response for bacterial survival in the pelagic marine environment. The goal of this investigation was to evaluate the relationship between chitinolytic gene expression and extracellular chitinase activity in individual cells of the marine bacterium Pseudoalteromonas sp. strain S91 attached to solid chitin. A green fluorescent protein reporter gene under the control of the chiA promoter was used to evaluate chiA gene expression, and a precipitating enzyme-linked fluorescent probe, ELF-97-N-acetyl-beta-D-glucosaminide, was used to evaluate extracellular chitinase activity among cells in the bacterial population. Evaluation of chiA expression and ELF-97 crystal location at the single-cell level revealed two physiologically distinct subpopulations of S91 on the chitin surface: one that was chitinase active and remained associated with the surface and another that was non-chitinase active and released daughter cells into the bulk aqueous phase. It is hypothesized that the surface-associated, non-chitinase-active population is utilizing chitin degradation products that were released by the adjacent chitinase-active population for cell replication and dissemination into the bulk aqueous phase.     

Differences in food webs and trophic states of Brazilian tropical humid and semi-arid shallow lakes: implications of climate change

Global warming may intensify eutrophication of shallow lakes by affecting nutrient loading, evaporation rates, and water level and thus produce major changes in food webs. We investigated to what degree food webs in tropical humid lakes differed from those in more eutrophic semi-arid lakes of the same latitude. Our results indicate that the catchment area-to-lake area ratio, nutrients, chlorophyll a, suspended solids, abundances of phytoplankton, zooplankton, and omnivorous fish as well as total fish catch per unit effort were all higher in the semi-arid lakes, whereas inlet water-to-evaporation ratio (proxy for water balance), water transparency, percentage macrophytes cover, and the piscivores:omnivores ratio were higher in the humid lakes. Our results suggest that reduced inlet water-to-evaporation ratio will increase lake eutrophication, which, in turn, as in temperate regions, will alter trophic structure of the freshwater community.

     

Plankton biomass partitioning in a eutrophic subtropical lake: comparison with results from temperate lake ecosystems

Plankton were sampled for 6 years in a subtropical eutrophiclake in FL, USA, and absolute and relative carbon biomass wasdetermined for bacteria, phytoplankton, heterotrophic and phototrophicnanoflagellates, ciliates, rotifers and crustacean zooplankton.We compared the results with findings from a comprehensive studyof carbon biomass partitioning in eutrophic German lakes withelucidate common patterns and differences. Similarities betweenthe temperate and subtropical systems included: similar seasonaldynamics, with maximal carbon biomass of nanoflagellates andmetazoan zooplankton in spring and phytoplankton in summer toautumn, yearly averaged carbon occurring mainly in the phytoplanktonand phytoplankton accounting for a much greater proportion ofcarbon than bacteria. There also were differences: the Floridalake had lower absolute and relative carbon biomass in crustaceanzooplankton, stronger dominance of protozoa in total grazercarbon biomass, a lower ratio of zooplankton to phytoplanktoncarbon and almost a monoculture of predation-resistant copepods(versus a relatively balanced distribution of carbon among cladocerans,copepods and rotifers in the temperate lakes). The subtropicallake also had 4-fold higher relative biomass of small filamentouscyanobacteria in its phytoplankton, which we attribute to lightlimitation. Although the Florida and German studies did notmeasure biomass of planktivorous fish, the differences observedhere are consistent with a recent hypothesis that fish predationexerts stronger top–down control on the pelagic food webin subtropical lakes than in temperate lakes of similar trophicstatus.     

Weak Response of Animal Allochthony and Production to Enhanced Supply of Terrestrial Leaf Litter in Nutrient-Rich Lakes

Ecosystems are generally linked via fluxes of nutrients and energy across their boundaries. For example, freshwater ecosystems in temperate regions may receive significant inputs of terrestrially derived carbon via autumnal leaf litter. This terrestrial particulate organic carbon (POC) is hypothesized to subsidize animal production in lakes, but direct evidence is still lacking. We divided two small eutrophic lakes each into two sections and added isotopically distinct maize litter to the treatment sections to simulate increased terrestrial POC inputs via leaf litter in autumn. We quantified the reliance of aquatic consumers on terrestrial resources (allochthony) in the year subsequent to POC additions by applying mixing models of stable isotopes. We also estimated lake-wide carbon (C) balances to calculate the C flow to the production of the major aquatic consumer groups: benthic macroinvertebrates, crustacean zooplankton, and fish. The sum of secondary production of crustaceans and benthic macroinvertebrates supported by terrestrial POC was higher in the treatment sections of both lakes. In contrast, total secondary and tertiary production (supported by both autochthonous and allochthonous C) was higher in the reference than in the treatment sections of both lakes. Average aquatic consumer allochthony per lake section was 27–40%, although terrestrial POC contributed less than about 10% to total organic C supply to the lakes. The production of aquatic consumers incorporated less than 5% of the total organic C supply in both lakes, indicating a low ecological efficiency. We suggest that the consumption of terrestrial POC by aquatic consumers facilitates a strong coupling with the terrestrial environment. However, the high autochthonous production and the large pool of autochthonous detritus in these nutrient-rich lakes make terrestrial POC quantitatively unimportant for the C flows within food webs.     

Zooplankton and environmental factors of a recovering eutrophic lake (Lysimachia Lake, Western Greece)

The present study investigates the zooplankton community dynamics and the abiotic environment in the eutrophic Lake Lysimachia (western Greece). The lake is considered to be recovering from eutrophication after the termination of an urban sewage inflow in 2000, and its waters are replenished constantly from the nearby oligotrophic Lake Trichonis. The results show that, although a decrease in nutrient concentrations was observed compared to the past, the lake still has eutrophic characteristics. This was reflected in the zooplankton community which is typical of those found in eutrophic lakes where rotifers prevail. Similarities among this lake and other nearby lakes were found considering the zooplankton community composition and seasonal variation. However, Lake Lysimachia is inhabited also by a number of different and even unique species (e.g., Moina micrura), suggesting that this ecosystem may be an important biodiversity refuge. Most of the zooplankton species were correlated with water temperature and, to a lesser extent, eutrophication key-water quality variables. Although there are few available data on the zooplankton of the lake, the abundance and composition of the community presenting characteristics indicative of intermediate trophic conditions and suggesting that the lake is probably under a kind of “biological” recovery.     

Summer zooplankton in small rivers in relation to selected conditions

The abundance and the biodiversity of summer zooplankton in the waters of the Drawa drainage (NW Poland) were studied, as was their relation to selected environmental conditions. The conditions upstream, especially in the outlets of lakes, did not affect the zooplankton communities downstream. This was also true of tributaries which had no influence on the shape of the zooplankton communities in the main river. The number of zooplankton in the outlets of eutrophic lakes was greater than in those of mesotrophic lakes. Increased vegetation cover significantly affected mainly the crustacean communities of zooplankton. Larger amounts of zooplankton were observed in rivers where the riparian zone was not covered with vegetation, but this difference was not significant. The hydrological conditions of the rivers and the Secchi depth visibility strongly impacted the composition of the zooplankton. The influence of abiotic factors was most pronounced on the abundance of cladocerans, and least pronounced on the abundance of rotifers.     

Aspergillus nidulans ChiA is a glycosylphosphatidylinositol (GPI)-anchored chitinase specifically localized at polarized growth sites

It is believed that chitinases play important physiological roles in filamentous fungi since chitin is one of the major cell wall components in these organisms. In this paper we investigated a chitinase gene, chiA, of Aspergillus nidulans and found that the gene product of chiA consists of a signal sequence, a region including chitinase consensus motifs, a Ser/Thr/Pro-rich region and a glycosylphosphatidylinositol (GPI)-anchor attachment motif. Phosphatidylinositol-specific phospholipase C treatment of the fusion protein of ChiA and enhanced green fluorescent protein (EGFP)-ChiA-EGFP-caused a change in its hydrophobicity, indicating that ChiA is a GPI-anchored protein. ChiA-EGFP localized at the germ tubes of conidia, at hyphal branching sites and hyphal tips. chiA expression was specifically high during conidia germination and in the marginal growth regions of colonies. These results suggest that ChiA functions as a GPI-anchored chitinase at the sites where cell wall remodeling and/or cell wall maturation actively take place.     

Do top-down and bottom-up controls interact to exclude nitrogen-fixing cyanobacteria from the plankton of estuaries? An exploration with a simulation model

Explaining the nearly ubiquitous absence of nitrogen fixation by planktonic organisms in strongly nitrogen-limited estuaries presents a major challenge to aquatic ecologists. In freshwater lakes of moderate productivity, nitrogen limitation is seldom maintained for long since heterocystic, nitrogen-fixing cyanobacteria bloom, fix nitrogen, and alleviate the nitrogen limitation. In marked contrast to lakes, this behavior occurs in only a few estuaries worldwide. Primary production is limited by nitrogen in most temperate estuaries, yet no measurable planktonic nitrogen fixation occurs. In this paper, we present the hypothesis that the absence of planktonic nitrogen fixers from most estuaries is due to an interaction of bottom-up and top-down controls. The availability of Mo, a trace metal required for nitrogen fixation, is lower in estuaries than in freshwater lakes. This is not an absolute physiological constraint against the occurrence of nitrogen-fixing organisms, but the lower Mo availability may slow the growth rate of these organisms. The slower growth rate makes nitrogen-fixing cyanobacteria in estuaries more sensitive to mortality from grazing by zooplankton and benthic organisms.We use a simple, mechanistically based simulation model to explore this hypothesis. The model correctly predicts the timing of the formation of heterocystic, cyanobacterial blooms in freshwater lakes and the magnitude of the rate of nitrogen fixation. The model also correctly predicts that high zooplankton biomasses in freshwaters can partially suppress blooms of nitrogen-fixing cyanobacteria, even in strongly nitrogen-limited lakes. Further, the model indicates that a relatively small and environmentally realistic decrease in Mo availability, such as that which may occur in seawater compared to freshwaters due to sulfate inhibition of Mo assimilation, can suppress blooms of heterocystic cyanobacteria and prevent planktonic nitrogen fixation. For example, the model predicts that at a zooplankton biomass of 0.2 mg l^–1, cyanobacteria will bloom and fix nitrogen in lakes but not in estuaries of full-strength seawater salinity because of the lower Mo availability. Thus, the model provides strong support for our hypothesis that bottom-up and top-down controls may interact to cause the absence of planktonic nitrogen fixation in most estuaries. The model also provides a basis for further exploration of this hypothesis in individual estuarine systems and correctly predicts that planktonic nitrogen fixation can occur in low salinity estuaries, such as the Baltic Sea, where Mo availability is greater than in higher salinity estuaries.     

The roles of the C-terminal domain and type III domains of chitinase A1 from Bacillus circulans WL-12 in chitin degradation.

The mature form of chitinase A1 from Bacillus circulans WL-12 comprises a C-terminal domain, two type III modules (domains), and a large N-terminal domain which contains the catalytic site of the enzyme. In order to better define the roles of these chitinase domains in chitin degradation, modified chiA genes encoding various deletions of chitinase A1 were constructed. The modified chiA genes were expressed in Escherichia coli, and the gene products were analyzed after purification by high-performance liquid chromatography. Intact chitinase A1 specifically bound to chitin, while it did not show significant binding activity towards partially acetylated chitosan and other insoluble polysaccharides. Chitinases lacking the C-terminal domain lost much of this binding activity to chitin as well as colloidal chitin-hydrolyzing activity. Deletion of the type III domains, on the other hand, did not affect chitin-binding activity but did result in significantly decreased colloidal chitin-hydrolyzing activity. Hydrolysis of low-molecular-weight substrates, soluble high-molecular-weight substrates, and insoluble high-molecular-weight substrates to which chitinase A1 does not bind were not significantly affected by these deletions. Thus, it was concluded that the C-terminal domain is a chitin-binding domain required for the specific binding to chitin and that this chitin-binding activity is important for efficient hydrolysis of the sufficiently acetylated chitin. Type III modules are not directly involved in the chitin binding but play an important functional role in the hydrolysis of chitin by the enzyme bound to chitin.     

Uncoupling of chlorophyll and nutrients in lakes – possible reasons,expected consequences

Total phosphorus and total nitrogen explained a low percentage of summer chlorophyll variability in epilimnia of the Great Masurian Lakes. Division of the whole data set into two subgroups of lakes improved approximation of the chlorophyll nutrient relationship but revealed also functional differences between the lakes distinguished in that way. Chlorophyll in eutrophic lakes correlated well with nitrogen and phosphorus, that in mesotrophic lakes (those with summer chlorophyll <=22 mg m^–3 as calculated in the model) was related to none of the nutrients. Higher summer chlorophyll content in epilimnetic waters was accompanied by higher chl:PP and chl:PN ratios. Algal adaptation to poor light conditions in eutrophic lakes is postulated as a possible reason for that difference.Chlorophyll – nutrient relationships varied with the trophic status of lakes. Epilimnetic chlorophyll strictly followed phosphorus changes in eutrophic lakes but did not do so in mesotrophic ones. Detailed comparison of selected meso- and eutrophic lakes showed marked differences in the seasonal changes of chlorophyll and nutrient concentrations and in sedimentation rates, especially in spring. Nutrient limitation rather than zooplankton grazing is suggested as a possible mechanism of controlling algal abundance and the sequence of spring events in a eutrophic lake. It is hypothesised that phosphorus turnover in eutrophic lakes is dominated by seasonal vertical fluxes, while in mesotrophic lakes it is more conservative with consumption and regeneration restricted mostly to metalimnion. Possible consequences of such conclusion are discussed in the paper.     

A matter of timing: heat wave impact on crustacean zooplankton

1. Climate change has affected zooplankton phenology and abundance in many freshwater ecosystems. The strong temperature anomalies that characterise summer heat waves make these events particularly suitable to study the effects of different seasonal warming patterns on zooplankton. Since heat waves are expected to occur more frequently under continuing climate change, they may also allow us to investigate how freshwater systems will be affected in the future. 2. Using a long‐term data set (1991–2007) from a shallow, eutrophic lake in Germany, we identify time periods in spring and summer during which cyclopoid copepods and bosminids are particularly sensitive to changes in water temperature. Based on this knowledge, we consider why summer populations responded differently to recent heat wave events that occurred at different times in the season. 3. Linear regressions of moving averages suggested that water temperatures shortly before and shortly after the clear‐water phase (CWP) were crucial for summer development of bosminids and cyclopoid copepods, respectively. Algal food availability (diatoms and cryptophytes) in the first weeks after the CWP also strongly influenced the summer populations of the two zooplankton groups. 4. Inter‐annual differences in water temperature during the critical time periods at least partly explained the contrasting responses of cyclopoid copepods and bosminids to heat wave events. 5. Our findings indicate that the zooplankton response to climate warming, particularly to heat wave events, is critically dependent on the temporal pattern of elevated water temperatures. Beyond that, we show that the summer zooplankton populations react to periods of warming in relation to events in the plankton annual cycle (such as the CWP in eutrophic lakes) rather than to warming at a fixed time in the season.     

The role of plankton, zoobenthos, and sediment in organic matter degradation in oligotrophic and eutrophic mountain lakes

Intensity of organic matter degradation, assessed by the respiratory electron transport system (ETS) activity, was studied in microplankton, zooplankton, chironomid larvae as the dominant group of the macrobenthos, and sediment in mountain lakes of different trophic levels in summer months. The highest ETS activities per unit of surface were observed in sediments. Significantly lower activities were observed in microplankton, and lower still in zooplankton, and chironomids. The total ETS activity m^–2 was higher in eutrophic lakes (Jezero na Planini pri Jezeru and Krnsko jezero) than in oligotrophic ones (Zgornje Kriko jezero, Spodnje Kriko jezero, Jezero v Ledvicah). The contributions of communities investigated to total ETS activity m^–2 differed between lakes of different trophic level. Estimation of respiratory carbon loss through different components revealed that the most of the organic matter was oxidized in sediments of mountain lakes. The respiratory carbon losses were higher through zooplankton than through microplankton in all lakes. Carbon losses through plankton components and sediments were significantly lower in oligotrophic than in eutrophic lakes. The contribution of respiratory carbon loss through chironomids to total carbon loss m^–2 was higher in oligotrophic than in eutrophic lakes. Therefore, it seems that contributions of microplankton and zooplankton to mineralization processes increase, and contributions of chironomids and sediment surface decrease with increasing trophic level of the lakes.     

Identification and characterization of the gene cluster involved in chitin degradation in a marine bacterium, Alteromonas sp. strain O-7.

Alteromonas sp. strain O-7 secretes chitinase A (ChiA), chitinase B (ChiB), and chitinase C (ChiC) in the presence of chitin. A gene cluster involved in the chitinolytic system of the strain was cloned and sequenced upstream of and including the chiA gene. The gene cluster consisted of three different open reading frames organized in the order chiD, cbp1, and chiA. The chiD, cbp1, and chiA genes were closely linked and transcribed in the same direction. Sequence analysis indicated that Cbp1 (475 amino acids) was a chitin-binding protein composed of two discrete functional regions. ChiD (1,037 amino acids) showed sequence similarity to bacterial chitinases classified into family 18 of glycosyl hydrolases. The cbp1 and chiD genes were expressed in Escherichia coli, and the recombinant proteins were purified to homogeneity. The highest binding activities of Cbp1 and ChiD were observed when alpha-chitin was used as a substrate. Cbp1 and ChiD possessed a chitin-binding domain (ChtBD) belonging to ChtBD type 3. ChiD rapidly hydrolyzed chitin oligosaccharides in sizes from trimers to hexamers, but not chitin. However, after prolonged incubation with large amounts of ChiD, the enzyme produced a small amount of (GlcNAc)(2) from chitin. The optimum temperature and pH of ChiD were 50 degrees C and 7.0, respectively.