Metal biosorption by two cyanobacterial mats in relation to pH, biomass concentration, pretreatment and reuse

The pH-dependent metal sorption by Oscillatoria- and Phormidium-dominated mats was effectively expressed by the Hill function. The estimated Hill functions can fruitfully predict the amount of metal sorbed at a particular initial pH. Pretreatment of biomass with 0.1 mmol L⁻¹ HCl was more effective than pretreatment with CaCl₂, HNO₃, NaOH, and SDS in enhancing metal sorption ability of the biomass. Desorption of metal ions in the presence of 100 mmol L⁻¹ HCl from metal-loaded mat biomass was completed within 1 h. After six cycles of metal sorption/desorption, sorption decreased by 6-15%. Only 6% and 11% of the biomass derived from the Oscillatoria sp.- and Phormidium sp.-dominated mats was lost during the cycling. The cyanobacterial mats seem to have better potential than several biomass types for use in metal sorption from wastewaters as they are ubiquitous, self-immobilized, and have good reusability.     

Phylogenetic relationships of nonaxenic filamentous cyanobacterial strains based on 16S rRNA sequence analysis

In order to determine the nearly complete 16S rRNA gene sequences of cyanobacteria originating from nonaxenic cultures, a cyanobacterium-specific oligonucleotide probe was developed to distinguish polymerase chain reaction (PCR) products of the cyanobacterial rRNA operons from those resulting from amplification of contaminating bacteria. Using this screening method the 16S rRNA genes of four nonaxenic filamentous cyanobacterial strains belonging to the generaLeptolyngbya andOscillatoria were cloned and sequenced. For the genusLeptolyngbya, the 16S rRNA sequence of the axenic strain PCC 73110 was also determined. Phylogenetic trees were constructed based on complete and partial sequences. The results show that the strainsLeptolyngbya foveolarum Komárek 1964/112,Leptolyngbya sp. VRUC 135 Albertano 1985/1, andLeptolyngbya boryanum PCC 73110 belong to the same cluster. StrainOscillatoria cf.corallinae SAG 8.92, which contains the rare photosynthetic pigment CU-phycoerythrin, is not closely related to other CU-phycoerythrin-containing cyanobacteria.Oscillatoria agardhii CYA 18, which is a representative of planktonicOscillatoria species that form toxic blooms in Norwegian inland waters, has no close relatives in the tree.     

Phylogenetic in situ/ex situ analysis of a sulfur mat microbial community from a thermal sulfide spring in the north Caucasus

A phylogenetic in situ/ex situ analysis of a sulfur mat formed by colorless filamentous sulfur bacteria in a thermal sulfide spring (northern spur of the main Caucasian ridge) was carried out. Nine phylotypes were revealed in the mat. Thiothrix sp. and Sphaerotilus sp. were the dominant phylotypes (66.3% and 26.3%, respectively). The 16S rRNA gene nucleotide sequence of Sphaerotilus sp. phylotype from the clone library was identical to the sequences of the seven Sphaerotilus strains isolated from the same source. A very high degree of similarity of Sphaerotilus strains revealed by ERIC-PCR fingerprints indicated little or no population diversity of this species in the mat. Thiothrix phylotype from the clone library and two Thiothrix strains isolated from the same mat sample differed in one to three nucleotides of 16S rRNA genes; this is an indication of this organism’s population variability in the mat. 16S rRNA genes of the strains and clones of Thiothrix sp. exhibited the highest similarity (ca. 99%) with Thiothrix unzii; the strains and clones of Sphaerotilus had 99% similarity with the type species Sphaerotilus natans (the only species of this genus) and therefore can be assigned to this species. The minor seven components belong to the phylotypes from the Proteobacteria (3%), as well as the Chlorobia, Cyanobacteria, Clostridia, and Bacteroidetes phylogenetic groups, each of them constituting not more than 1%. Intracellular accumulation of elemental sulfur by Sphaerotilus similar to other filamentous sulfur bacteria was demonstrated for the first time (both in the population of the sulfur spring and in cultures with sulfide). Although mass growth of Sphaerotilus and Thiothrix is typical of bacterial populations of anthropogenic ecosystems (the activated sludge of treatment facilities), stable communities of these bacteria have not been previously found in the sulfur mats or “threads” of natural sulfide springs.     

Ecology of Potamanthus myops (Walsh) (Ephemeroptera: Potamanthidae) in a Michigan stream (USA)

The ecology of Potamanthus myops (Potamanthidae) was examined for a one year period in a central Michigan fourth order stream. Lowest nymphal densities (4.7 m^-2) occurred in mid-summer after adult emergence, while highest densities were found during late summer and fall (68.0 m^-2 and 66.2 m^-2) after the new cohort had hatched. Biomass was the inverse of density with highest values, 67.7 mg m^-2, just prior to peak adult emergence in June and lowest in late summer (16 mg m^-2). Nymphs maintained a contagious spatial distribution with k values ranging between 0.19 and 1.40. P. myops nymphs fed mostly on detritus (96.8%), placing them in the collector-gatherer functional group. Nymphs were always found in erosional zones associated with rock substrate. Nymphs used their mandibular tusks for positioning themselves beneath rocks, while also appearing to use them for defensive purposes.     

Chemical reaction- and particle diffusion-based kinetic modeling of metal biosorption by a Phormidium sp.-dominated cyanobacterial mat

The present study explores the suitability of chemical reaction-based and diffusion-based kinetic models for defining the biosorption of Cu(II), Cd(II) and Pb(II) by Phormidium sp.-dominated mat. The time-course data of metal sorption by the test mat significantly (r² = 0.932-0.999) fitted to the chemical reaction-based models namely pseudo-first-order, -second-order, and the general rate law. However, these models fail to accurately describe the kinetics of metal biosorption due either to prefixed order or unjustifiable change in rate constant and reaction order with varying concentrations of metal and biomass in the solution. The diffusion-based models, namely, the intra-particle diffusion model and the external mass transfer model fitted well to the time-course metal sorption data, thus suggesting involvement of both external and intra-particle diffusion processes in sorption of test metals by mat biomass. However, the Boyd kinetic expression clearly showed that the external mass transfer is the dominant process.     

人工遮光和营养添加对河流反硝化活性和反硝化细菌群落结构的影响

河流生态系统受到人类活动例如河岸带森林植被毁损和农业活动施肥等的干扰日益加剧,而这些活动使河流接收的光照增多、河流的氮磷营养盐浓度增加。微生物的反硝化作用是河流去除氮的有效途径。在汉江的一级支流金水河上游核心保护区内选取6条溪流开展野外控制实验,利用营养添加模拟河流中营养的增加,遮盖河面来模拟源头溪流的隐蔽状态,来研究河流沉积物中微生物的反硝化作用对光照和营养改变的响应,并利用高通量测序(Mi Seq)技术研究在两种处理下河流沉积物中nir S型反硝化细菌的群落结构变化。结果显示:营养元素添加促进了沉积物的反硝化活性,河面遮盖抑制了沉积物的反硝化活性。营养添加和遮盖两种处理均降低了控制实验区域内脱氯单胞菌属(优势菌属)的相对丰度,同时也降低了该区域nir S型反硝化菌群落的Chao多样性。本研究初步证实了光照增加和河流的营养增加提高了河流沉积物反硝化活性,并为提高河流的脱氮能力提供科学依据。     

Ecology of a saline stream: community responses to spatial gradients of environmental conditions

Spatial changes in structural and functional characteristics of fish and macroinvertebrate communities in eastern Kentucky were investigated in a drainage system chronically exposed to high levels of chloride salts from nearby oilfield operations. Salinity levels at biological monitoring stations ranged from 0.12–31.3‰. Lotic regions with salinities greater than 10‰ were dominated by larvae of the dipterans Ephydra and Culicoides. In regions with salinities less than 10‰ species richness increased more or less linearly with decreasing levels of chloride salts. Ephemeropterans appeared to be one of the major invertebrate groups least tolerant of elevated NaCl levels and were absent in regions with salinities greater than 2‰ Availability of food resources, such as periphyton and particulate organic matter, did not appear to be grossly altered in disturbed regions, and it is suggested that the observed distribution of macroinvertebrate fauna was largely in response to taxonomic differences in salt tolerance. Fish seemed to be more tolerant of highly saline conditions, and several species were observed in regions experiencing salinities as high as 15‰. Accordingly, assemblages of fish taxa along the salinity gradient may have been influenced by trophic factors, such as spatial limitations in availability of invertebrate prey.     

Interactions of the bacterial assemblages of a prairie stream with dissolved organic carbon from riparian vegetation

The zonation of riparian vegetation on the Konza Prairie Research Natural Area of Kansas (tallgrass prairie ecosystem) provided a situation where the influences on uptake rates of the different DOC qualities could be measured in the field. Leachates from grasses disappeared rapidly from stream water in in situ chambers at upstream sites in grassland reaches and at downstream sites in forested reaches. The reverse observation, uptake of leachates from trees, revealed that DOC disappeared rapidly from chambers placed in the forested reaches but bacteria in the grassland reaches did not utilize tree leachate as rapidly. The pattern varies seasonally. The interpretation of these observations was aided by aseptic laboratory culture on the different substrate types and by measurements in poisoned chambers to evaluate abiotic uptake.     

Life history of a lowland burrowing mayfly, Ephemera orientalis (Ephemeroptera: Ephemeridae), in a Korean stream

Life history aspects of Ephemera orientalis, a common lowland burrowing mayfly that resides in temperate East Asia, were studied in terms of voltinism, secondary production, and accumulated degree days for larval development. From March 1998 to June 1999, larvae were sampled monthly (weekly or biweekly during the emergence period) from a lower reach of the Gapyeong stream in Korea, a stream typical of temperate East Asia, using a Surber sampler (0.25 m², mesh 0.25 mm). As a result, the mean density of E. orientalis was 47.21 ± 13.58 indiv. m⁻² during the study period. Very small larvae less than 2 mm in body length were sampled on three separate occasions, and emergence was observed between late April and early October, except during late May. Based on the larval body length distribution and emergence time, two different developmental groups could be distinguished: the slow developmental group (S-group) and the fast developmental group (F-group). The F-group completed its life cycle within 4 months, whereas the S-group had a one-year life cycle. The S-group could be divided into two subgroups, dubbed the S1 and S2-groups, based on larval development. The developmental groups alternate their life cycles in the order: S1→F→S2→S1. Estimated annual production of the larvae was 68.81 mg DW m⁻² y⁻¹; mean biomass was 8.43 mg DW m⁻²; the annual production to mean biomass ratio was 8.16. The annual mean water temperature of the study year was 14.76 ± 6.63°C. The total accumulated degree days for larval development was 1396°C for the F-group, 2,055°C for the S1-group, and 1,975°C for the S2-group. Two different adult body size groups were distinguished (P < 0.001 by t-test): larger adults, which belonged to the S-group, were present throughout the emergence period, whereas smaller adults, which belonged to the F-group, were mainly present later in the emergence period (August–October). The difference in the accumulated degree days between the developmental groups may explain the size differences of adults.     

Mobilifilum chasei: Morphology and ecology of a spirochete from an intertidal stratified microbial mat community

Spirochetes were found in the lower anoxiphototrophic layer of a stratified microbial mat (North Pond, Laguna Figueroa, Baja California, Mexico). Ultrastructural analysis of thin sections of field samples revealed spirochetes approximately 0.25 m in diameter with 10 or more periplasmic flagella, leading to the interpretation that these spirochetes bear 10 flagellar insertions on each end. Morphometric study showed these free-living spirochetes greatly resemble certain symbiotic ones, i.e., Borrelia and certain termite spirochetes, the transverse sections of which are presented here. The ultrastructure of this spirochete also resembles Hollandina and Diplocalyx (spirochetes symbiotic in arthropods) more than it does Spirochaeta, the well known genus of mud-dwelling spirochetes. The new spirochete was detected in mat material cellected both in 1985 and in 1987. Unique morphology (i.e., conspicuous outer coat of inner membrane, large number of periplasmic flagella) and ecology prompt us to name a new free-living spirochete.This article is dedicated to the memory of our dear friend and colleague David G. Chase (1935–1987)     

In situ nutrient assays of periphyton growth in a lowland Costa Rican stream

Nutrient limitation of primary production was experimentally assessed using an in situ bioassay technique in the Quebrada Salto, a third-order tropical stream draining the northern foothills of the Cordillera Central in Costa Rica. Bioassays employed artificial substrata enriched with nutrients that slowly diffuse through an agar-sand matrix (Pringle & Bowers, 1984). Multiple comparisons of regression coefficients, describing chlorophyll-a accrual through time for different nutrient treatments, revealed positive micronutrient effect(s). Micronutrient treatment combinations (Fe, B, Mn, Zn, Co, Mo, EDTA), supplemented with and without nitrate and phosphate, exhibited significantly greater chlorophyll-a accrual over all other treatments (P < 0.05), supporting over three times that of the control after 14-d of substratum colonization. Neither of the major nutrients (N or P) produced a significant stimulation, although the N treatment displayed 50% more chlorophyll-a than the control after 14-d. Similarly, Si, EDTA, and Si + N + P treatments did not exhibit chlorophyll-a response curves that were significantly different from the control. During the experiment, mean NH₄-N and (NO₂ + NO₃)-N concentrations in the Salto were 2.0 µM (28.6 µg · l^–1) and 7.2 µM (100.2 µg · l ^–1), respectively. High concentrations of PO₄-P ( = 2.0 µM; 60.9 µg · l^–1) and TP ( = 3.0 µM; 94.0 µg · l^–1) were also found, and consequently low molar N:P ratios = 4.7). Despite the potential for N limitation in the system, both N and P appear to be at growth saturating levels. This may be due to micronutrient limitation and/or light limitation of periphyton growth in densely shaded upstream portions of the stream.     

Seasonal changes in composition and biomass of epiphytic algae on the macrophyte Ranunculus penicillatus in a chalk stream,with estimates of production,and observations on the epiphytes of Cladophora glomerata

The epiphytic algae on surfaces of the macrophyte Ranunculus penicillatus (Dumort.) Bab. var. calcareus (R. W. Butcher) C. D. K. Cook and on Cladophora glomerata (L.) Kütz growing in the River Itchen at Otterbourne near Southampton were studied between February 1984 and June 1985. The river at this site has a mean flow rate of 0.33 m s^–1, and is about 16 m wide and on average 20 cm deep, with a discharge ranging through the year between 0.34 and 2.46 m³ s^–1. The pH of the river varies little around 8.2, with a mean alkalinity of 236 mg HCO _inf3 ^sup1 l^–1, because of its origin from chalk springs. Ranunculus grows throughout the year, with peaks of biomass in spring and autumn. It forms a very large surface for attachment of epiphytes, and covers on average 40% of the stream bed. Numbers of epiphyte cells removed from Ranunculus ranged through the year between 52 × 10³ and 271 × 10³ cells mm^–2 stream floor, with maximum numbers in April, and a secondary peak in October. This pattern partly reflects fluctuations in the biomass of Ranunculus; the number of cells per unit area of plant surface showed a broader spring peak and lower fluctuations in other seasons. Diatoms formed 65 to 98% of these epiphyte cells, with chlorophytes reaching their peak (10%) in summer and cyanophytes (25%) in autumn. Estimates of biomass of these epiphytic forms, derived from measurements of chlorophyll c, indicate a range between 30 and 100 g dry weight m^–2 of weed bed. Colonisation studies showed that the algae grow and reproduce throughout the year, with a mean generation time of about 5 days, suggesting an annual production of about 3 kg dry weight m^–2 of weed bed, which makes epiphytic algae the principal primary producers in the stream. The numbers and biomass of epiphytic algae on Cladophora are considerably less.The species of epiphytic algae found on Ranunculus were generally similar to those growing on Cladophora, and to epilithic algae on pebbles of the stream bed, but different species were dominant on the different substrata. Algal cells in the water column were all derived from benthic habitats, although their relative abundance was very different.     

Distribution of benthic algae and macroinvertebrates along a thermal stream gradient

The distribution and abundance of benthic algae and macroinvertebrates were examined along a natural thermal gradient formed by hot springs in Little Geysers Creek, Sonoma Co., California, USA. Maximum water temperatures ranged from 52 °C at the uppermost station to 23 °C at a station 400 m downstream. Benthic chlorophyll a decreased exponentially from 2.5 g m^–2 at 52 °C to less than 0.1 g m^–2 at 23 °C, a pattern of decline also exhibited by algal phaeophytin. Blue-green algae dominated at higher temperatures but were replaced by filamentous green algae and diatoms at lower temperatures.Macroinvertebrates were absent at temperatures 45 °C; the highest density (> 150 000 m^–2, mainly Chironomidae) occurred at 34 °C, whereas biomass was highest (4.6 g m^–2, as dry weight) at 23 °C and species richness (15 species) was highest at 27 °C. The two predominant macroinvertebrate populations (the midge Tanytarsus sp. and the caddisfly Helicopsyche borealis) occurred at sites that were several degrees below their lethal thermal threshold, suggesting that a temperature buffer is maintained.     

Recovery of the macroinvertebrate community below a wastewater treatment plant input in a Mediterranean stream

We sampled chlorophyll a, benthic organic matter, and benthic macroinvertebrates in June 2001 in La Tordera stream (Catalonia, NE Spain), receiving a wastewater treatment plant (WWTP) input. Samples were collected in six equidistant transects in three reaches located upstream (UP), few m below (DW1), and 500 m below the WWTP input (DW2). Our first objective was to assess the effects of the point source on the structure and functional organization of the benthic macroinvertebrate community. Our second objective was to determine if the self-purifying capacity of the stream implied differences between the communities of the DW1 and the DW2 reaches. The WWTP input highly increased discharge, nutrient concentrations, and conductivity and decreased dissolved oxygen. At the DW1 and the DW2 reaches, taxa richness, EPT taxa (Ephemeroptera, Plecoptera, and Trichoptera), and Shannon diversity decreased and gatherer relative density increased relative to the UP reach. At the UP reach, CPOM and FPOM standing crops were similar, whereas at the DW1 and the DW2 reaches CPOM was two times higher than FPOM. Detailed analysis showed that major changes in the benthic community occurred abruptly between 80 and 90 m downstream of the point source (middle of the DW1 reach). At this location, chlorophyll a concentration, density of macroinvertebrates, taxa richness, and scraper relative density increased, whereas gatherer relative percentage decreased. The macroinvertebrate community at the DW2 reach was comparable to that at the second middle of the DW1 reach (DW1B). The macroinvertebrate community at the DW1B and the DW2 reaches were quite similar to that at the UP reach, indicating that the recovery capacity of the stream from nutrient enrichment was high.     

Bacterial community shift along a subsurface geothermal water stream in a Japanese gold mine

Change of bacterial community occurring along a hot water stream in the Hishikari gold mine, Japan, was investigated by applying a combination of various culture-independent techniques. The stream, which is derived from a subsurface anaerobic aquifer containing plentiful CO₂, CH₄, H₂, and NH₄⁺, emerges in a mine tunnel 320 m below the surface providing nutrients for a lush microbial community that extends to a distance of approximately 7 m in the absence of sunlight-irradiation. Over this distance, the temperature decreases from 69°C to 55°C, and the oxidation-reduction potential increases from –130 mV to +59 mV. In the hot upper reaches of the stream, the dominant phylotypes were: 1) a deeply branching lineage of thermophilic methane-oxidizing -Proteobacteria, and 2) a thermophilic hydrogen- and sulfur-oxidizing Sulfurihydrogenibium sp. In contrast, the prevailing phylotypes in the middle and lower parts of the stream were closely related to ammonia-oxidizing Nitrosomonas and nitrite-oxidizing Nitrospira spp.. Changes in the microbial metabolic potential estimated by competitive PCR analysis of genes encoding the enzymes, particulate methane monooxygenase (pmoA), ammonia monooxygenase (amoA), and putative nitrite oxidoreductase (norB), also substantiated the community shift indicated by 16S rRNA gene analysis. The diversity of putative norB lineages was assessed for the first time in the hot water environment. Estimation of dominant phylotypes by whole-cell fluorescent in situ hybridization and changes in inorganic nitrogen compounds such as decreasing ammonium and increasing nitrite and nitrate in the mat-interstitial water along the stream were consistent with the observed transition of the bacterial community structure in the stream.     

Life cycle phenology of common detritivores from a temperate rainforest stream

The timing of life cycles, including growth rates, was determined for eight common species of detritivorous insects in a second-order stream in southwestern British Columbia, Canada. Six of the species (Zapada cinctipes, Z. haysi, Malenka californica, M. cornuta, Capnia sp., and Lepidostoma roafi) had simple, univoltine life cycles. The leuctrid stonefly Despaxia augusta has a 2-year life cycle, with an apparent egg diapause of about 6 months. The chironomid Brillia retifinis produced at least three generations per year. The major growth periods for the set of species considered here span the entire year. Adults of several species exhibited seasonal declines in size at emergence, but one species had larger adults as the emergence period proceeded. Closely related taxa had more similar life cycle timing than more distantly related species suggesting a degree of phylogenetic constraint in phenology of their life cycles. The influence of the timing of leaf drop on timing of life cycles for these animals does not fit with proposed scenarios based on fast and slow leaf processing rates.     

The spring energy budget of the algal mat community in a Crimean hypersaline lake determined by microcalorimetry

The energy contents (standing stock) of the floating mat formed by the green alga Cladophora sivaschensis and the energy transfers through it were quantified for a shallow hypersaline lake (at Cape Khersones, Crimea, Ukraine) during the spring months. Appropriate direct calorimetric techniques were applied to: (i) measure the heat energy dissipated by the mat community and by the free bacterioplankton in the water column below it; and (ii) differentiate between the heat flows by the heterotrophic and the phototrophic components of the community. It was shown that Cladophora biomass reached a peak of 579.5 g C m^–2, contributing more than 99.6% of the total mat community. Throughout the spring, the total bacterial energy transfer (6 to 23 mW m^–2) was as little as 1.1 to 2.6% of the total heat dissipated by the microplankton community. The rest of the estimated heat energy (584 to 1488 mW m^–2) was associated with Cladophora metabolism. In the spring community: (i) the rate of biomass accumulation in the lake photic layer significantly exceeded its heterotrophic mineralisation; (ii) the efficiency of the microbial loop was too low to process even a minor part of the accumulated organic matter. The microcalorimetric technique was shown to be a highly promising approach for further studies of natural microbial mats and biofilms, biological systems with complex metabolism that involves not only aerobic processes but also anaerobic catabolism under local hypoxic/microxic conditions.     

16S rRNA sequences reveal uncultured inhabitants of a well-studied thermal community

Abstract Molecular methods are beginning to reveal inhabitants of natural microbial communities which have nerver before been cultured. Our approach involves selective cloning of naturally occurring 16S rRNA sequences as cDNA, and comparison of these sequences to a database which includes 16S rRNA sequences of isolated community members. We provide here an overview of the method and its potential for community analysis. A 16S rRNA sequence retrieved from the well-studied hot spring cyanobacterial mat in Octopus Spring (Yellowstone National Park) is shown as an example of one contributed by an uncultured member of the community.