Differential partitioning of geosmin and 2-methylisoborneol between cellular constituents in Oscillatoria tenuis

A newly isolated strain of Oscillatoria tenuis produces geosmin and 2-methylisoborneol (MIB) simultaneously. The intra- and extracellular distribution of both compounds was studied. Geosmin was much less released to the medium compared to MIB. Different amounts of each of the compounds were in different cellular fractions. The geosmin and MIB in these fractions were recoverable by ammonium sulphate precipitation, suggesting binding to protein. Thylakoid and cytoplasmic membranes contained most geosmin while high MIB protein ratios were found in a colourless protein fraction as obtained by gel exclusion chromatography.Abbreviations MIB 2-methylisoborneol - SDS sodium dedecyl sulphate     

Identification and expression analysis of the gene associated with geosmin production in Lyngbya kuetzingii UTEX 1547 (cyanobacteria)

Cyanobacteria are the major producers of geosmin in natural waters. To identify a gene involved in geosmin biosynthesis in cyanobacteria, the polymerase chain reaction (PCR) was used to amplify a 2298-bp open reading frame (ORF) from the geosmin-producing cyanobacterium Lyngbya kuetzingii UTEX 1547. This ORF encoded a protein of 765 amino acids. Alignment of the deduced amino acid sequence demonstrated that geoL had high similarity to the corresponding genes of Oscillatoria sp. PCC 6506 (100% identity), Calothrix sp. PCC 7507 (89%), Anabaena ucrainica CHAB 1432 (88%), A. ucrainica CHAB 2155 (87%), Nostoc punctiforme PCC 73102 (87%), Phormidium sp. P2r (84%) and Cylindrospermum stagnale PCC 7417 (83%), and modest similarity to myxobacteria (61–73%). It also indicated geoL with low similarity to the corresponding genes of actinomycetes (<60%). The encoded protein GEOL was estimated to have two geosmin synthase domains, and each contained two strictly conserved Mg²⁺-binding motifs (aspartate-rich motif and NSE triad). The geoL gene was shown to be responsible for geosmin biosynthesis in L. kuetzingii UTEX 1547. Then, geoL had been cloned into pET21a(+) vector and expressed in Escherichia coli BL21(DE3) with the isopropyl-β-d-thiogalactoside (IPTG) induction. The recombinant GEOL protein was purified and exhibited a single band (M_W ∼ 90 kDa) on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which was consistent with the predicted molecular weight (M_W) of 87,046 Da. In conclusion, this study has confirmed that geosmin synthase gene and its expression product can be identified and characterized from cyanobacteria, which will help understand the fundamental biological mechanism of geosmin biosynthesis in cyanobacteria.     

原核生物中土霉味化合物二甲萘烷醇和2-甲基异茨醇生物合成研究进展

由原核生物蓝藻和放线菌引起的水体土霉异味问题备受世界的关注。本文以土霉味化合物二甲萘烷醇和2-甲基异茨醇的研究历史为主线,综述了土霉味化合物二甲萘烷醇和2-甲基异茨醇的化学特性、生物合成途径以及生物合成相关基因和酶等方面的研究进展,探讨了土霉味化合物二甲萘烷醇和2-甲基异茨醇生物合成研究的重要意义,并展望了土霉味化合物二甲萘烷醇和2-甲基异茨醇生物合成研究存在的问题及发展的方向。     

Effects of light and temperature on the odor production of 2-methylisoborneol-producing Pseudanabaena sp. and geosmin-producing Anabaena ucrainica (cyanobacteria)

Frequent off-flavor events caused by geosmin and 2-methylisoborneol (MIB) have attracted research on the main producers, cyanobacteria. This study evaluated the effects of light and temperature on the odor production of MIB-producing Pseudanabaena sp. Lauterborn and geosmin-producing Anabaena ucrainica (Schhorb.) Watanabe. The maximum MIB production and lowest growth rate (indicated by the chlorophyll a (Chl a)) were observed at 35 °C compared with that at 10 °C and 25 °C. Cultures grown under a light intensity of 60 μmol photons m⁻² s⁻¹ demonstrated the highest MIB production and minimum growth rate, whereas the minimum MIB production and maximum growth rate were obtained under 10 μmol photons m⁻² s⁻¹. Similar patterns were observed for geosmin production. A. ucrainica had the highest geosmin production and lowest Chl a concentration under 10 °C and 60 μmol photons m⁻² s⁻¹. Moreover, greater proportions of geosmin and MIB were released into extracellular under growth-inhibiting temperatures and light intensities. An inverse correlation between odor production and the cell growth rate was suggested, and this relationship may reflect the competition for substrates of odor and Chl a synthesis. Thus, the accumulation of geosmin and MIB was probably the result of decreased cellular metabolic activity in cyanobacteria.     

Species richness and assemblage structure of Trichoptera in Danish streams

• 1 Species richness and assemblages of Trichoptera from 157 ‘pristine’ Danish lowland stream sites were analyzed, for dependence on geographical position of the sites and simple physical variables, using two way indicator species analysis and detrended correspondence analysis.
• 2 A total of 106 species were recorded, representing ≈ 90% of the species pool known from Danish streams. Only seven species occurred at more than half the sites, whereas an additional 11 species were found at more than a quarter of the sites.
• 3 Although sites showed significant regional differences in environmental variables (stream order, width, slope and presence/absence of riparian forest), species richness and assemblages were primarily correlated with stream order, width and slope. Maximum richness was found at the largest (5th order) stream sites.
• 4 Regional differences in species assemblages were found, with several species being absent from the islands Funen and Bornholm. Species assemblages also differed between forested and non‐forested stream sites.
• 5 We found evidence that stream temperature may be of only minor importance in determining Trichoptera species richness and assemblage composition in Danish streams compared to other size‐related physical factors.

     

Effect of pH on microbial degradation of leaf litter in seven streams of the English Lake District

Summary Rates of degradation of alder, oak and grass leaf packs with associated microbial populations were measured in seven streams pH 6.8–4.9. Streams were chosen from upland and lowland sites of the same river for contrasts in pH, water chemistry and riparian vegetation. The most important factor governing rates of degradation is the physical and chemical nature of the leaf material. At pH 6.8 rates of degradation, k, and microbial colonization were higher than at pH 5.5: k on alder x6; on oak x2; on grass x2. At lowland sites, pH 6.8, higher decay rates were associated with high levels of microbial colonization including c.14 spp of aquatic hyphomycete fungi—regardless of riparian vegetation. Decay rates were similar at upland sites, pH 6.8 and 6.6, involving high levels of colonization by fewer fungal species and fewer bacteria—regardless of riparian vegetation-though grass was barely degraded at upland sites of any pH. At pH 5.5, slow decay rates were associated with low levels of microbial colonization and few fungal species. Largest microbial populations at low pH associated with riparian trees did not lead to markedly increased decay rates. Factors of water chemistry at low pH appear to inhibit microbial metabolism. The implications of these findings for stream invertebrates active in the winter is discussed.     

Macrophyte diversity and composition in relation to substratum characteristics in regulated and unregulated Danish streams

1. The objective of the present study was to examine how the physical stream environment in regulated and unregulated lowland streams affects the diversity and distribution of macrophyte communities. We analysed the abundance, distribution and composition of macrophytes, together with physical parameters, in seven regulated and seven unregulated unshaded Danish stream reaches. 2. Total macrophyte coverage was similar in the regulated and unregulated streams, but species richness and Shannon diversity were higher in the unregulated streams. Overall, we found fifty-two different species in the regulated stream reaches and sixty-two in the unregulated stream reaches. The spatial distribution of macrophytes on the stream bottom was more heterogeneous in the unregulated streams. 3. We found positive correlations between the coverage and diversity of macrophytes and the coverage of coarse-textured substratum types on the stream bottom, as well as between macrophyte coverage and diversity and substratum heterogeneity. We also found that the macrophytes were more heterogeneously distributed where substratum heterogeneity was greater. 4. The species growing both submerged and emergent were more abundant in the regulated streams, whereas species growing only submerged were more abundant in the unregulated streams. Species growing submerged, species growing both submerged and emergent, and species only growing emergent segregated differently in a canonical correspondence analysis ordination. The submerged species were primarily associated with coarser-textured substrata, whereas species growing both submerged and emergent, and species growing only emergent were associated with finer-textured substrata. 5. The most abundant species in the regulated streams, Sparganium emersum, accounting for almost one-third of the total macrophyte coverage, was primarily associated with clay and sandy bottom substrata, whereas the most abundant species in the unregulated streams, Batrachium peltatum, was primarily associated with gravel and stony substrata.     

Effects of temperature and light on the growth and geosmin production of Lyngbya kuetzingii (Cyanophyta)

The effects of temperature and light on the growth and geosmin production of Lyngbya kuetzingii were determined. Of the three temperatures tested, 10, 25 and 35°C, the maximal geosmin concentration and geosmin productivity were yielded at 10°C, while the highest chl a production was observed at 25°C. In the studies on light intensity, the maximal geosmin concentration and geosmin productivity were observed at 10 μmol m⁻² s⁻¹, while the highest chl a production was at 20 μmol m⁻² s⁻¹. It was suggested that more geosmin was synthesized with lower chl a demand. Meanwhile, the relative amounts of extra- and intracellular geosmin were investigated. Under optimum growth conditions (20 μmol m⁻² s⁻¹, 25°C; BG-11 medium), the amounts of extracellular geosmin increased as the growth progressed and reached the maximum in the stationary phase, while the intracellular geosmin reached its maximum value in the late exponential phase, and then began to decline. However, under the low temperature (10°C) or light (10 μmol m⁻² s⁻¹) conditions, more intracellular geosmin was synthesized and mainly accumulated in the cells. The proportions of extracellular geosmin were high, to 33.33 and 32.27%, respectively, during the stationary phase at 35°C and 20 μmol m⁻² s⁻¹. It was indicated that low temperature or light could stimulate geosmin production and favor the accumulation of geosmin in cells, while more intracellular geosmin may be released into the medium at higher temperatures or optimum light intensity.     

Long-term effects of stream management on plant communities in two Danish lowland streams

Submerged macrophytes grow abundantly in most shallow streams common in the cultivated lowlands of northwestern Europe. Weed-cutting has been practised for years in many of these streams to reduce the risk of flooding of adjacent land. Our objective was to quantify long-term impacts of weed-cutting on macrophyte communities in two Danish rivers. We found that the total macrophyte coverage was similar in the weed-cut and uncut reaches in the two rivers, but species richness, diversity and patch complexity were higher in the uncut reaches. The spatial distribution of macrophytes on the stream bottom was also more heterogeneous in the uncut stream reaches. We also found evidence of a strong effect of weed-cutting on macrophyte species composition. P. natans was abundant in the uncut reaches in both streams but practically eliminated in the cut reaches, despite the fact that its basic habitat requirements were met. Also, the established phase strategy of the macrophyte community was affected by weed-cutting. Species displaying characteristically ruderal traits were more abundant in the cut reaches and species with competitive abilities were only abundant in the uncut stream reaches. We suggest that important species traits in streams, where the weed is cut regularly, are associated with rapid growth and high dispersal-capacity. Our results indicate that weed-cutting can contribute significantly to a decline in species diversity in streams. To provide optimal conditions for diverse stream macrophyte communities, we therefore suggest that weed-cutting should be minimised.     

Riparian ecotones and spatial variation of fish assemblages in Portuguese lowland streams

The first results of a long-term study on the role of riparian ecotones on the population and community dynamics of Iberian stream fish are presented and discussed. Riparian and macrophyte cover, bank slope and depth were among the most important variables affecting fish distribution. In general small fish favoured shallow areas with high macrophyte cover, whereas large fish dominated in deep areas with a high riparian cover. Slight spatial changes in terrestrial prey use were found suggesting a minor role for this resource during autumn. Finally, no significant spatial differences were found for linear growth, although some differences were obtained for the condition factor.     

First report of geosmin and 2-methylisoborneol (2-MIB) in Dolichospermum and Oscillatoria from Vietnam

Limnology - Cyanobacteria are the major producers of the taste and odour compounds geosmin and 2-methylisoborneol (2-MIB) in drinking water and fish worldwide. In recent years, the outbreaks of...     

Temporal variability in fish assemblages from disturbed and undisturbed streams

Fish assemblages were sampled by electrofishingover a two- to ten-year period in undisturbedand anthropogenically disturbed South Carolinacoastal plain streams. Jaccard similarity,Bray–Curtis similarity, and Spearman rankcorrelations among samples collected from thesame sites over time were significantly greaterat undisturbed sites than at disturbed sites,suggesting greater fish assemblage persistenceand stability at the undisturbed sites. TheIndex of Biotic Integrity (IBI) also exhibitedsignificantly less variation over time atundisturbed sites than at disturbed sites.Physical habitat structure changed more overtime at disturbed sites than at undisturbedsites, and this variability was directlyrelated to temporal variability in fishassemblage structure. Comparisons betweenmultiple and single pass electrofishing samplessuggested that only a small proportion of thetemporal variability observed at the studysites was caused by inefficient sampling.Assessment of temporal variation in fishassemblage structure can serve as an indicatorof environmental disturbance and facilitate thedistinction of substantive ecological changefrom normal background variation.     

Assessing effects of water abstraction on fish assemblages in Mediterranean streams

1. Water abstraction strongly affects streams in arid and semiarid ecosystems, particularly where there is a Mediterranean climate. Excessive abstraction reduces the availability of water for human uses downstream and impairs the capacity of streams to support native biota. 2. We investigated the flow regime and related variables in six river basins of the Iberian Peninsula and show that they have been strongly altered, with declining flows (autoregressive models) and groundwater levels during the 20th century. These streams had lower flows and more frequent droughts than predicted by the official hydrological model used in this region. Three of these rivers were sometimes dry, whereas there were predicted by the model to be permanently flowing. Meanwhile, there has been no decrease in annual precipitation. 3. We also investigated the fish assemblage of a stream in one of these river basins (Tordera) for 6 years and show that sites more affected by water abstraction display significant differences in four fish metrics (catch per unit effort, number of benthic species, number of intolerant species and proportional abundance of intolerant individuals) commonly used to assess the biotic condition of streams. 4. We discuss the utility of these metrics in assessing impacts of water abstraction and point out the need for detailed characterisation of the natural flow regime (and hence drought events) prior to the application of biotic indices in streams severely affected by water abstraction. In particular, in cases of artificially dry streams, it is more appropriate for regulatory agencies to assign index scores that reflect biotic degradation than to assign ‘missing’ scores, as is presently customary in assessments of Iberian streams.     

The relative importance of shading and nutrients on algal production in subtropical streams

1. We used artificial substrata in forested and open streams in South-East Queensland, Australia, to determine the relative importance of shading from riparian vegetation and of nutrients on periphyton growth, and whether nitrogen and/or phosphorus limited algal productivity.
2. Nutrient-diffusing substrata consisting of agar enriched with N, P and N + P, and controls without nutrients, were deployed in duplicate at 15 sites in headwater streams with riparian canopy cover ranging from 0 to 88%.
3. Shading was the over-riding factor controlling periphyton biomass accrual on the artificial substrata, with nutrients playing a relatively minor role. Microscopic examination of periphyton scrapings taken after 7 weeks revealed that diatoms dominated on the artificial substrata in shaded streams, whereas filamentous green algae dominated the algal assemblage in the more open canopy streams.
4. Whilst nutrients had little effect on the accrual of algal biomass compared with riparian shading, there was evidence that nitrogen, and not phosphorus, stimulated periphyton production in streams with sufficient light.     

Production and degradation of geosmin in a stratified lake with anaerobic hypolimnion (Schleinsee)

Abstract Depth profiles of geosmin concentrations were determined over the year in a stratified lake with anaerobic hypolimnion. Two independent sources of geosmin were observed. Most geosmin was produced under anaerobic conditions in the hypolimnion in the autumn. A much smaller source of geosmin production was actinomycetes in the epilimnion. With the onset of the autumnal circulation of the water body, rapid aerobic degradation of geosmin was observed.     

Primary production and potential availability of autochthonous carbon in southern Ontario streams

Primary production was measured in forested and open streams in southern Ontario to identify the main factors influencing algal growth, and to test the assumption that autochthonous inputs are negligible in forested headwater streams. In low order streams in southern Ontario, primary production is related primarily to light, temperature, and substrate, and appears to be largely independent of location on the river continuum. Net primary production on rock averaged 2.24 and 0.098 g Cm^–2 day^–1 during the summer at meadow and forested sites, respectively, showing peaks in the spring and fall at the forested sites. Net primary production on sediment was significant, averaging 20% (range of 6 to 82%) of net primary production on rock. When adjusted for assumed invertebrate assimilation efficiency, autochthonous production in riffles at forested sites could provide an estimated 30% of total carbon inputs potentially available to aquatic invertebrates.     

An index of biotic integrity based on fish assemblages for subtropical streams in southern Brazil

Several methods for calculating indices of biological integrity (IBIs) have been developed for different ecogeographic regions of the world. All of them calculate IBI scores by comparing against reference sites or historical data on fish assemblage composition. Because of intensive agriculture and urbanization in our study area, we located no reference sites, and historical information about fish assemblage composition did not exist. Instead, we developed hypothetical reference scores based on seasonal electrofishing surveys at six study sites in adjacent but geomorphologically different watersheds. Our IBI included 10 metrics that varied with degree of degradation. We found that Shannon–Wiener indices varied significantly between seasons, indicating significant changes in species composition. Therefore, we calculated seasonal IBI scores also, but these did not significantly differ from each streams general IBI score, which was calculated on the basis of four samples per site. General stream IBI scores reflected differing levels of anthropogenic disturbance.     

Leaf litter breakdown, microbial respiration and shredder production in metal-polluted streams

1. If species disproportionately influence ecosystem functioning and also differ in their sensitivities to environmental conditions, the selective removal of species by anthropogenic stressors may lead to strong effects on ecosystem processes. We evaluated whether these circumstances held for several Colorado, U.S.A. streams stressed by Zn. 2. Benthic invertebrates and chemistry were sampled in five second–third order streams for 1 year. Study streams differed in dissolved metal concentrations, but were otherwise similar in chemical and physical characteristics. Secondary production of leaf‐shredding insects was estimated using the increment summation and size‐frequency methods. Leaf litter breakdown rates were estimated by retrieving litter‐bags over a 171 day period. Microbial activity on leaf litter was measured in the laboratory using changes in oxygen concentration over a 48 h incubation period. 3. Dissolved Zn concentrations varied eightfold among two reference and three polluted streams. Total secondary production of shredders was negatively associated with metal contamination. Secondary production in reference streams was dominated by Taenionema pallidum. Results of previous studies and the current investigation demonstrate that this shredder is highly sensitive to metals in Colorado headwater streams. Leaf litter breakdown rates were similar between reference streams and declined significantly in the polluted streams. Microbial respiration at the most contaminated site was significantly lower than at reference sites. 4. Our results supported the hypothesis that some shredder species contribute disproportionately to leaf litter breakdown. Furthermore, the functionally dominant taxon was also the most sensitive to metal contamination. We conclude that leaf litter breakdown in our study streams lacked functional redundancy and was therefore highly sensitive to contaminant‐induced alterations in community structure. We argue for the necessity of simultaneously measuring community structure and ecosystem function in anthropogenically stressed ecosystems.     

淡水鱼池土腥异味物质含量与浮游藻类和放线菌生物量的关系

通过顶空固相微萃取-气质联用色谱测定北京市精养鱼池中两种主要土腥异味物质(土臭味素和二甲基异莰醇)含量,同时测定鱼池中浮游藻类和放线菌生物量,研究了土腥异味物质含量与浮游藻类和放线菌生物量之间的关系。结果表明,试验鱼池中土腥异味物质以土臭味素为主,土臭味素在精养鱼池中普遍存在,含量为1.22～35.58ng·L-1,二甲基异莰醇在部分鱼池中被检出,含量1.39～6.00ng·L-1。精养鱼池中共检出浮游藻类6门22属,生物量17.33～178.34mg·L-1,以硅藻和裸藻为主。放线菌共测到4个属,其中链霉菌Streptomyces sp.是主要种类,放线菌总生物量0～76×104ind.L-1。鱼池中浮游藻类总生物量与土臭味素含量正相关。浮游藻类中的颗粒直链藻Melosira granulata和条纹小环藻Cyclotella striata可能是北京地区夏秋季节淡水精养鱼池中能够产生土臭味素的主要藻类,裸藻和其他鞭毛藻类对池中异味化合物的产生可能作用较小。