Abundance of picophytoplankton in the halocline of a meromictic lake, Lake Suigetsu, Japan

Numerous (0.5 to 4.8 × 10⁵ cells/ml), small phytoplankton (smaller than 0.5–1 × 1–2 μm in cell size, picophytoplankton) were distributed in the halocline (depth 2–12 m, 4–14 practical salinity units) of the saline meromictic lake, Lake Suigetsu (35°35′ N, 135°52′ E), located in the central part of the coast of Wakasa Bay along the Japan Sea in Fukui Prefecture, Japan. Vertical distribution of phytoplankton revealed that the maximum number of picophytoplankton was always observed near or a little deeper than the oxic-anoxic boundary layer (depth 5–6 m); they were dominant phytoplankton in the water layer deeper than the oxic-anoxic boundary from July to late September 2005. Spectral analysis of autofluorescence emitted from the particle fractions smaller than 5 μm measured with a spectrofluorometer and from individual cells measured with a microscope photodiode array detector revealed that the major component of picophytoplankton was phycoerythrin-rich, unicellular cyanobacteria (picocyanobacteria). Eukaryotic phytoplankton about 2.5 μm in diameter were also found, but the numbers were low. Fluorescence intensity of chlorophyll a at 685 nm (room temperature) emitted from the particle fractions smaller than 5 μm was increased by the addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. These observations indicated that at least some picophytoplankton had a functional photosystem II in the halocline where sulfide, the potential inhibitor of oxygenic photosynthesis, was always present. The large abundance together with their physiological potency suggest that picophytoplankton are one of the important primary producers in the halocline of Lake Suigetsu. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

Dominance of green sulfur bacteria in the chemocline of the meromictic Lake Suigetsu, Japan, as revealed by dissimilatory sulfite reductase gene analysis

This study investigated the spatiotemporal abundance and diversity of the α-subunit of the dissimilatory sulfite reductase gene (dsrA) in the meromictic Lake Suigetsu for assessing the sulfur-oxidizing bacterial community. The density of dsrA in the chemocline reached up to 3.1 × 10⁶ copies ml^?1 in summer by means of quantitative real-time PCR and it was generally higher than deeper layers. Most of the dsrA clones sequenced were related to green sulfur bacteria such as Chlorobium phaeovibrioides, C. limicola, and C. luteolum. Below the chemocline of the lake, we also detected other dsrA clones related to the purple sulfur bacterium Halochromatium salexigens and some branching lineages of diverse sequences that were related to chemotrophic sulfur bacterial species such as Magnetospirillum gryphiswaldense, Candidatus Ruthia magnifica, and Candidatus Thiobios zoothamnicoli. The abundance and community compositions of sulfur-oxidizing bacteria changed depending on the water depth and season. This study indicated that the green sulfur bacteria dominated among sulfur-oxidizing bacterial population in the chemocline of Lake Suigetsu and that certain abiotic environmental variables were important factors that determined sulfur bacterial abundance and community structure.     

Dominant bacterioplankton populations in the meromictic Lake Suigetsu as determined by denaturing gradient gel electrophoresis of 16S rRNA gene fragments

Lake Suigetsu is a typical meromictic lake in Japan characterized by a permanent chemocline at a depth of between 3 and 8 m separating the oxic freshwater mixolimnion from anoxic saline sulfidogenic monimolimnion. Dominant bacterioplankton populations in Lake Suigetsu were investigated using PCR-denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments. The bacterial population was vertically stratified, and temporal shifts in the microbial communities were observed in both the oxic and anoxic layers of Lake Suigetsu during the sampling period. Several dominant DGGE bands were excised and sequenced. In the chemocline, green sulfur bacteria phylogenetically related to the genera Prosthecochloris, Pelodyctyon, and Chlorobium within the phylum Chlorobi were dominant; the colorless sulfur bacteria closely related to the genus Thiomicrospira were detected. These sulfur bacterial groups appear to be important in the biogeochemical cycling of sulfur and/or carbon in Lake Suigetsu. Bacterial sequences affiliated with the Bacteroidetes phylum were frequent among the dominant fragments in the DGGE profiles throughout the water column. Populations possessing a fermentative metabolism exist in Bacteroidetes, suggesting they may contribute to the degradation of organic matter in the anoxic environment of Lake Suigetsu.     

Depth distribution of microbial diversity in Mono Lake,a meromictic soda lake in California

We analyzed the variation with depth in the composition of members of the domain Bacteria in samples from alkaline, hypersaline, and currently meromictic Mono Lake in California. DNA samples were collected from the mixolimnion (2 m), the base of the oxycline (17.5 m), the upper chemocline (23 m), and the monimolimnion (35 m). Composition was assessed by sequencing randomly selected cloned fragments of 16S rRNA genes retrieved from the DNA samples. Most of the 212 sequences retrieved from the samples fell into five major lineages of the domain BACTERIA: alpha- and gamma-Proteobacteria (6 and 10%, respectively), Cytophaga-Flexibacter-Bacteroides (19%), high-G+C-content gram-positive organisms (Actinobacteria; 25%), and low-G+C-content gram-positive organisms (Bacillus and Clostridium; 19%). Twelve percent were identified as chloroplasts. The remaining 9% represented beta- and delta-Proteobacteria, Verrucomicrobiales, and candidate divisions. Mixolimnion and oxycline samples had low microbial diversity, with only 9 and 12 distinct phylotypes, respectively, whereas chemocline and monimolimnion samples were more diverse, containing 27 and 25 phylotypes, respectively. The compositions of microbial assemblages from the mixolimnion and oxycline were not significantly different from each other (P = 0.314 and 0.877), but they were significantly different from those of chemocline and monimolimnion assemblages (P < 0.001), and the compositions of chemocline and monimolimnion assemblages were not significantly different from each other (P = 0.006 and 0.124). The populations of sequences retrieved from the mixolimnion and oxycline samples were dominated by sequences related to high-G+C-content gram-positive bacteria (49 and 63%, respectively) distributed in only three distinct phylotypes, while the population of sequences retrieved from the monimolimnion sample was dominated (52%) by sequences related to low-G+C-content gram-positive bacteria distributed in 12 distinct phylotypes. Twelve and 28% of the sequences retrieved from the chemocline sample were also found in the mixolimnion and monimolimnion samples, respectively. None of the sequences retrieved from the monimolimnion sample were found in the mixolimnion or oxycline samples. Elevated diversity in anoxic bottom water samples relative to oxic surface water samples suggests a greater opportunity for niche differentiation in bottom versus surface waters of this lake.     

Methane production in meromictic Ace Lake,Antarctica

Methane occurred in the monimolimnion, at depths greater than 11 m, of an antarctic meromictic lake, Ace Lake (depth 24.7 m). Although the water of the lake was of approximate marine salinity, bottom waters were depleted in sulfate (less than 1 mmol 1^–1). The temperature of the bottom waters of the lake were constantly between 1 °C and 2 °C. Rates of methanogenesis from ¹⁴C-labelled precursors (bicarbonate, formate and acetate) were determined in time course experiments with the detection of ¹⁴CH₄ produced by a gas chromatography-gas proportional counting system. Rates of ¹⁴CH₄ production were difficult to determine as the reactions were always near our limit of detection.Reliable determinations of rates of methanogenesis at some depths using some precursors were obtained, the fastest rate being 2.5 µmol kg^–1 day^–1 at depth 20 m. Assuming constant rates of methanogenesis with time, this would equate to a turnover of methane in the lake every two years.The slow rate of methanogenesis suggests that the methanogens in Ace Lake may be working at well below their optimum temperature although definitive statements regarding the presence of psychrophilic methanogens in this antarctic lake must await isolation attempts or longer field studies using alternative methodologies.     

Biogeochemical processes in the saline meromictic Lake Kaiike, Japan: implications from molecular isotopic evidences of photosynthetic pigments

Stable carbon and nitrogen isotopic compositions were determined for individual photosynthetic pigments isolated and purified from the saline meromictic Lake Kaiike, Japan, to investigate species-independent biogeochemical processes of photoautotrophs in the natural environment. In the anoxic monimolimnion and benthic microbial mats, the carbon isotopic compositions of BChls e and isorenieratene related to brown-coloured strains of green sulfur bacteria are substantially ( approximately 10 per thousand) depleted in (13)C relative to those found in the chemocline. In conjunction with 16S rDNA evidence reported previously, it strongly suggests that Pelodyctyon luteolum inhabited and photosynthesized in the anoxic monimolimnion and benthic microbial mats by using (13)C-depleted regenerated CO(2). By contrast, both Chl a and BChl a in the monimolimnion and microbial mats have similar isotopic compositions as they do in the chemocline, implying that the source organisms live only in the chemocline. In the chemocline, the nitrogen isotopic compositions of BChl e homologues ranges from -7.7 to-6.5 per thousand, whereas that of BChl a is -2.1 per thousand. These isotopic compositions suggest that green sulfur bacteria Chlorobium phaeovibrioides would conduct nitrogen fixation in the chemocline, whereas purple sulfur bacteria Halochromatium sp. and cyanobacteria Synechococcus sp. may assimilate nitrite.     

Distribution of chloropigments in suspended particulate matter and benthic microbial mat of a meromictic lake, Lake Kaiike, Japan

We investigated the distribution of chloropigments in a small meromictic lake, Lake Kaiike, south-west Japan. In the water-column, concentrations of Chl a related to cyanobacteria, BChl a related to purple sulphur bacteria, and three types of BChl e homologues (BChls e1, e2 and e3) related to brown-coloured green sulphur bacteria, were maximal at the redox boundary. Below the redox boundary, absolute concentrations of Chl a and BChl a gradually decreased with depth, whereas BChls e remained rather constant. Suspended particulate matter (SPM) at the deeper region of the anoxic water-column was enriched in highly alkylated BChl e homologues compared with SPM at the redox boundary. The shift in the relative content of highly alkylated BChl e homologues beneath the boundary was associated with community related adaptation of brown-coloured green sulphur bacteria to changes in light quality/quantity, resulting from the optical absorption and reflectance of SPMs in the overlying water-column. Benthic microbial mats were characterized by high abundances of BChls e, in which highly alkylated homologues were substantially abundant. This suggests that the BChls e in the microbial mat may be derived from the low-light adapted brown-coloured green sulphur bacteria forming the bacterial mat.     

Anoxygenic phototrophic bacteria of the high-altitude meromictic Lake Gek-Gel,Azerbaijan

The anoxygenic phototrophic bacterial community of the high-altitude meromictic Lake Gek-Gel (Azerbaijan) was investigated in September 2003. The highest concentration of bacteriochlorophyll e (48 μg/l) was detected at a depth of 30 m; the peak of bacteriochlorophyll a (4.5 μg/l) occurred at 29 m. Phylogenetic analysis revealed that brown-colored green sulfur bacteria Chlorobium phaeobacteroides predominated in the lake. Nonsulfur purple bacteria phylogenetically close to Blastochloris sulfoviridis were found in insignificant amounts; these organisms have not been previously reported in Lake Gek-Gel.     

Densities and distribution of flagellates and ciliates in the chemocline of saline,meromictic Lake Shunet (Siberia,Russia)

The vertical and seasonal distributions of the phytoflagellate Cryptomonas spp., and its most common, the planktonic ciliate predators (Oligotrichida, Scuticociliatida, Hypotrichida and Prostomatida) were investigated in chemocline region of small saline, meromictic lake Shunet (Siberia, Russia) during 2003 and 2005. The lake has a pronounced chemocline, with abundance of purple and green sulphur bacteria. Vertical distribution of the Cryptomonas populations near the oxic/anoxic boundary layer was studied at close intervals in water sampled using a hydraulically operated thin-layer sampler. In both summer and winter, Cryptomonas peaked in water stratum 5–10 cm above anoxic zone or in the anoxic zone water column in the chemocline (about 5 m). Ciliate densities and biomass were also much higher in chemocline than in mixolimnion. The range of diurnal migration of Cryptomonas population was not very wide, and it was restricted to layers with high light intensity. The ciliates were sometimes detected above the upper border of the anoxic zone but also several centimetres below this zone.     

Spatial and temporal distribution of archaeal diversity in meromictic,hypersaline Ocnei Lake (Transylvanian Basin,Romania)

Saline, meromictic lakes with significant depth are usually formed as a result of salt mining activity. Ocnei Lake is one of the largest Transylvanian (Central Romania) neutral, hypersaline lake of man-made origin. We aimed to survey the seasonal dynamics of archaeal diversity in the water column of Ocnei Lake by employing microbiological methods as well as molecular techniques based on the sequence analysis of the 16S rRNA gene. We found that archaeal diversity in the water column increased with depth and salinity, with 8 OTUs being detected in the epilimnion compared to 21 found in the chemocline, and 32 OTUs in the monimolimnion. Down to 3.5 m depth, the archaeal community was markedly dominated by the presence of an unclassified archaeon sharing 93 % sequence identity to Halogeometricum spp. At the chemocline, the shift in archaeal community composition was associated with an increase in salinity, the main factor affecting the vertical distribution of archaeal assemblages. It appears that the microoxic and hypersaline monimolimnion is populated by several major haloarchaeal taxa, with minor fluctuations in their relative abundances throughout all seasons. The culturable diversity was reasonably correlated to the dominant OTUs obtained by molecular methods. Our results indicate that Ocnei Lake represents a relatively stable extreme habitat, accommodating a diverse and putatively novel archaeal community, as 30 % of OTUs could not be classified at the genus level.     

Dominant microbial composition and its vertical distribution in saline meromictic Lake Kaiike (Japan) as revealed by quantitative oligonucleotide probe membrane hybridization

Vertical distributions of dominant bacterial populations in saline meromictic Lake Kaiike were investigated throughout the water column and sediment by quantitative oligonucleotide probe membrane hybridization. Three oligonucleotide probes specific for the small-subunit (SSU) rRNA of three groups of Chlorobiaceae were newly designed. In addition, three general domain (Bacteria, Archaea, and Eukarya)-specific probes, two delta-Proteobacteria-specific probes, a Chlorobiaceae-specific probe, and a Chloroflexi-specific probe were used after optimization of their washing conditions. The abundance of the sum of SSU rRNAs hybridizing with probes specific for three groups of Chlorobiaceae relative to total SSU rRNA peaked in the chemocline, accounting for up to 68%. The abundance of the delta-proteobacterial SSU rRNA relative to total SSU rRNA rapidly increased just below the chemocline up to 29% in anoxic water and peaked at the 2- to 3-cm sediment depth at ca. 34%. The abundance of SSU rRNAs hybridizing with the probe specific for the phylum Chloroflexi relative to total SSU rRNA was highest (31 to 54%) in the top of the sediment but then steeply declined with depth and became stable at 11 to 19%, indicating the robust coexistence of sulfate-reducing bacteria and Chloroflexi in the top of the sediment. Any SSU rRNA of Chloroflexi in the water column was under the detection limit. The summation of the signals of group-specific probes used in this study accounted for up to 89% of total SSU rRNA, suggesting that the DGGE-oligonucleotide probe hybridization approach, in contrast to conventional culture-dependent approaches, was very effective in covering dominant populations.     

Temporal and vertical distribution of planktonic rotifers in a meromictic crater lake,Lake Nyahirya (Western Uganda)

Quantitative zooplankton sampling was carried out biweekly during 16 months in Lake Nyahirya, a Western Uganda crater lake. Six out of 24 rotifer species recorded from the lake were quantitatively important (Horaella brehmi, Brachionus angularis, B. caudatus, Keratella tropica, Filinia longiseta and F. opoliensis, and three others were common (Brachionus falcatus, Asplanchna sieboldi and Conochiloides natans). Rotifer peaks were correlated with rainy periods and relatively clear water, which coincide with the European spring and autumn. All species were able to deal with hypoxic conditions. A clear niche separation can be observed between different genera and between species of the same genus. The population dynamics of the rotifers seem to be ruled by repeated irregular environmental fluctuations. Rainfall appears to be a primary steering factor.     

Spatial distribution and seasonal changes of pesticides in Lake Biwa,Japan

The spatial distribution and seasonal variation in the concentrations in Lake Biwa of pesticides used in paddy fields were studied. Lake Biwa is the largest lake in Japan and is a recognized water resource for 14 million people in the Kinki district. Samples were collected nine times from April to December 2001 at ten sites within the lake and at the mouths of six influent rivers. Weekly sampling was also carried out at a single site on an effluent river. Among the 20 pesticides analyzed, the detection frequencies in surface water were almost 100% for simetryn, bromobutide, and isoprothiolane; around 75% for molinate and pyroquilon; around 30% for three herbicides and one fungicide; and almost zero for the remaining substances. The maximum concentrations of pesticides detected frequently in the lake were in the range 0.1–0.4µgl^–1. The occurrence of a few pesticides below the thermocline may be explained by thermal stratification and vertical circulation. Although the thermocline suppressed vertical diffusion in spring and summer during pesticide application periods, a few pesticides remaining at the surface of the lake in winter were transported to the hypolimnion by vertical circulation and remained there even after the reestablishment of the thermocline. The half-lives of pesticides in the lake were estimated to be more than a year for simetryn, half a year for bromobutide, 1.5 months for molinate, and 1 month for dimepiperate. The main cause of elimination for molinate and dimepiperate was estimated to be degradation, that for simetryn was outflow, and for bromobutide both degradation and outflow were significant.     

Seasonal and spatial community dynamics in the meromictic Lake Cadagno

The seasonal and spatial variations in the community structure of bacterioplankton in the meromictic alpine Lake Cadagno were examined by temporal temperature gradient gel electrophoresis (TTGE) of PCR-amplified 16S rDNA fragments. Two different amplifications were performed, one specific for the domain Bacteria (Escherichia coli positions 8-536) and another specific for the family Chromatiaceae (E. coli positions 8-1005). The latter was followed by semi-nested reamplification with the bacterial primer set, allowing comparison of the two PCR approaches by TTGE. The TTGE patterns of samples from the chemocline and the anoxic monimolimnion were essentially identical, whereas the oxic mixolimnion displayed distinctively different banding patterns. For samples from the chemocline and the monimolimnion, dominant bands in the Bacteria-specific TTGE profiles comigrated with bands obtained by the semi-nested PCR approach specific for Chromatiaceae. This observation suggested that Chromatiaceae are in high abundance in the anoxic water layer. All dominant bands were excised and sequenced. Changes in the community structure, as indicated by changes in the TTGE profiles, were observed in samples taken at different times of the year. In the chemocline, Chomatium okenii was dominant in the summer months, whereas Amoebobacter purpureus populations dominated in autumn and winter. This change was confirmed by fluorescent in situ hybridization.     

Spatiotemporal shift in sulphide concentration in hypolimnic water column in Lake Hiruga,a saline lake in Japan

A 2-year study was designed to investigate the spatiotemporal distribution of sulphide in the water column of a saline lake, Lake Hiruga in Fukui, Japan. Water samples were collected in Lake Hiruga periodically between April 2014 and March 2016 at appropriate depths with simultaneous in situ measurements of physicochemical parameters such as temperature, salinity and dissolved oxygen (DO) concentration. Sulphide started accumulating in the hypolimnion in September 2014, whilst that did in June 2015. The sulphide concentrations increased toward the end of December in both 2014 and 2015. Then the sulphide disappeared from the hypolimnion in April 2015 and February 2016. The difference in starting season of the sulphide accumulation was closely connected with DO concentrations in the hypolimnion in circulation periods of winter season. The DO concentrations were regulated by water column stability before thermal stratification periods. Our analysis of the sulphide distribution for the 2 years of the study suggested that accumulation of sulphide in the hypolimnion during stratified periods appears to be associated with strength of water column stability in the winter-spring seasons before stagnant periods in Lake Hiruga.     

Anaerobic microbial communities in Lake Pavin, a unique meromictic lake in France

The Bacteria and Archaea from the meromictic Lake Pavin were analyzed in samples collected along a vertical profile in the anoxic monimolimnion and were compared to those in samples from the oxic mixolimnion. Nine targeted 16S rRNA oligonucleotide probes were used to assess the distribution of Bacteria and Archaea and to investigate the in situ occurrence of sulfate-reducing bacteria and methane-producing Archaea involved in the terminal steps of the anaerobic degradation of organic material. The diversity of the complex microbial communities was assessed from the 16S rRNA polymorphisms present in terminal restriction fragment (TRF) depth patterns. The densities of the microbial community increased in the anoxic layer, and Archaea detected with probe ARCH915 represented the largest microbial group in the water column, with a mean Archaea/Eubacteria ratio of 1.5. Terminal restriction fragment length polymorphism (TRFLP) analysis revealed an elevated archaeal and bacterial phylotype richness in anoxic bottom-water samples. The structure of the Archaea community remained rather homogeneous, while TRFLP patterns for the eubacterial community revealed a heterogeneous distribution of eubacterial TRFs.     

滴水湖叶绿素a时空分布及其与水质因子的关系

根据2011年9月-2012年8月监测数据,分析上海市人工湖泊——滴水湖叶绿素a的时空分布及其与主要环境因子的相互关系.结果表明:滴水湖叶绿素a含量随时间的变化幅度较大,表现为3、7月出现高峰而其他月份较低;叶绿素a含量在空间分布上具有一定的分异性,湖心普遍偏高,其他各点相对较低;对滴水湖叶绿素a采用主成分多元线性回归分析(APCS-MLR),得到描述水质因子的潜变量F1、F2与其相关性较大,建立了潜变量与叶绿素a含量的相关性模型;滴水湖叶绿素a含量与溶解氧(DO)、水温、总溶解磷(TDP)、活性磷(AP)有着密切的相关性;磷为滴水湖水体初级生产力的限制因子;因此,磷是有效控制滴水湖水体富营养化的关键因子.     

Annual cycle of phytoplankton in Ace Lake,an ice covered,saline meromictic lake

The annual cycle of phytoplankton in saline, meromictic Ace Lake (68°2S.4S, 78°11.1E) in the Vestfold Hills, Antarctica, was studied from January, 1979 to January 1980. Ace Lake has permanent gradients of temperature, salinity, dissolved oxygen, and hydrogen sulphide, and is ice covered with up to 2 m of ice for 10–12 months each year. The phytoplankton community had low diversity, consisting of only four species, all flagellates — a prasinophyte Pyramimonas gelidicola McFadden et al., a cryptophyte of the genus Cryptomonas; an unidentified colourless microflagellate, and an unarmoured dinoflagellate. These were restricted to the oxic zone of the lake from the surface to 10 m.The phytoplankton had a cycle of seven months of active growth over spring and summer. Low numbers of cells survived in the water column over winter. Spring growth was initiated below the ice by increased light penetration through the ice into the lake, enhanced at the time by the removal of surface snow which accumulated on the ice over winter. Peak phytoplankton biomass production was by the shade adapted P. gelidicola and occurred at the interface of the oxic and anoxic zones where substantial available nitrogen as ammonia is found.The three dominant phytoplankton species displayed distinct vertical stratification over the oxic zone. This stratification was not static and developed over spring as the flagellates migrated to preferred light climate zones. Mean cell volume of two of the flagellates varied significantly over the year. Minimum volumes were recorded in winter and volume increased progressively over spring to reach maximum mean cell volume in summer. Mean cell volume was positively correlated with light intensity (maximum ambient PAR at the respective depth for date of sample). Low cell volume in winter may be related to winter utilization of carbohydrate reserves by slow respiration, and may represent a survival mechanism.