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1.
The distribution of phospholipid ester-linked fatty acids (PLFA) in sediments of eutrophic bays (Hiroshima Bay and Aki Nada) was studied to quantify the microbial biomass, community structure, and nutritional status. A total of 63 fatty acids in the range of C10 to C24 were determined. They consist of saturated fatty acids, branched fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids, and variation was revealed in the relative proportions of these fatty acids in sediments. On the basis of the PLFA concentration in sediments, the calculated microbial biomass showed variation (mean ± standard deviation = 0.70 × 108 ± 0.53 × 108 cells per g [dry weight] of sediment) in the eutrophic bays. In sediments, a higher amount of biomass was observed in the coastal area of Hiroshima Bay than that observed in the rest of the bay and adjacent Aki Nada. The microbial community structure of the present study area, as characterized by the PLFA profiles, showed very low percentages of polyunsaturated fatty acids and long-chain fatty acids characteristic of microeukary-otes and terrestrial input, respectively, and high percentages of fatty acids characteristic of bacteria. The distribution of PLFA profiles also showed the relative contribution of both aerobic and anaerobic bacteria, especially sulfate-reducing bacteria, in the study area. The relative proportions of PLFA revealed distinctive differences among the stations of the study area, as is evidenced from six clusters obtained for the PLFA profiles. The results of Tukey's honestly significant difference test further confirmed that the sediments in the coastal area of Hiroshima Bay were significantly enriched by a number of fatty acids when compared with other areas investigated where relatively few fatty acids were present in significant quantities. No marked variation in environmental parameters in the surface- and bottom-water samples was observed, indicating the absence of any water movement in the study area. Furthermore, low redox potential and the levels of sulfide in the sediment revealed the reduced condition of the sediment. The existing environmental conditions and pollution of the study area were attributed to the observed microbial community structure in the sediments.  相似文献   

2.
Sediment samples were collected monthly from Acton Lake, a eutrophic reservoir located in an agricultural region of southwestern Ohio, from three stations (River, Middle, and Dam) during the period May 1995 through January 1997. Sedimentary microbial biomass and community structures from these stations were studied using phospholipid analysis. At the River and Middle stations, the water column remained aerobic throughout the year, whereas the water overlying the Dam station sediments became anaerobic during summer stratification. Sedimentary microbial biomass at the River and Middle stations, as measured by the phospholipid phosphate (PLP) method, ranged from 225 to 450 nmol PLP g?1 d.w. (dry weight). Sedimentary microbial biomass at the Dam station was typically greater and ranged from 500 to 1,500 nmol PLP g?1 d.w. Principal component analysis of phospholipid fatty acid (PLFA) profiles indicated that the sedimentary microbial communities at all three stations displayed seasonal patterns of change. Among these patterns of change was a shift from aerobic microorganisms during times of cold water to anaerobic microorganisms during times of warm water. The Dam station differed from the River and Middle stations in that sediments from this station had disproportionately more polyenoic fatty acids, whereas sediments from the River and Middle stations had disproportionately more bacterial fatty acids. These data suggest that the Dam station may be a depositional zone for microeukaryotic phytoplankton produced in the overlying water column. These findings have implications for the understanding of carbon flux in reservoirs and preservation of organic matter in aquatic systems.  相似文献   

3.
Twenty-eight sediment samples collected from Osaka Bay, Japan, were analyzed for phospholipid ester-linked fatty acids (PLFA) to determine regional differences in microbial community structure of the bay. The abundance of three major groups of C10 to C19 PLFA (saturated, branched, and monounsaturated PLFA), which accounted for 84 to 97% of the total PLFA, indicated the predominance of prokaryotes in the sediment. The distribution of six clusters obtained by similarity analysis in the bay revealed a marked regional distribution in the PLFA profiles. Total PLFA concentrations (0.56 to 2.97 μg/g [dry weight] of the sediment) in sediments also showed marked variation among the stations, with higher concentrations of total PLFA in the central part of the bay. The biomass, calculated on the basis of total PLFA concentration, ranged from 0.25 × 108 to 1.35 × 108 cells per g (dry weight) of the sediment. The relative dominance of microbial groups in sediments was described by using the reported bacterial biomarker fatty acids. Very small amounts of the characteristic PLFA of microeukaryotes in sediments indicated the restricted distribution of microeukaryotes. By examining the distribution of clusters and groups of microorganisms in the bay, there were two characteristics of the distribution pattern: (i) the predominance of anaerobic bacteria and gram-positive prokaryotes, characterized by the high proportions of branched PLFA in the eastern and northeastern sides of the bay, where the reported concentrations of pollutants were also high, and (ii) the predominance of aerobic prokaryotes and eukaryotes, except for a few stations, in the western and southwestern sides of the bay, as evidenced by the large amounts of monounsaturated PLFA. Such significant regional differences in microbial community structure of the bay indicate shifts in microbial community structure.  相似文献   

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6.
Fatty acids are a widely studied group of lipids of sufficient taxonomic diversity to be useful in defining microbial community structure. The extraordinary resolution of glass capillary gas-liquid chromatography can be utilized to separate and tentatively identify large numbers of fatty acid methyl esters derived from the lipids of estuarine detritus and marine benthic microbiota without the bias of selective methods requiring culture or recovery of the microbes. The gas-liquid chromatographic analyses are both reproducible and highly sensitive, and the recovery of fatty acids is quantitative. The analyses can be automated, and the diagnostic technique of mass spectral fragmentation analysis can be readily applied. Splitless injection on glass capillary gas chromatographic columns detected by mass spectral selective ion monitoring provides an ultrasensitive and definitive monitoring system. Reciprocal mixtures of bacteria and fungi, when extracted and analyzed, showed progressive changes of distinctive fatty acid methyl esters derived from the lipids. By manipulating the environment of an estuarine detrital microbial community with antibiotics and culture conditions, it was possible to produce a community greatly enriched in eucaryotic fungi, as evidenced by scanning electron microscopic morphology. The fatty acid methyl esters from the lipids in the fungus-enriched detritus showed enrichment of the C18 dienoic and the C18 and C20 polyenoic esters. Manipulation of the detrital microbiota that increased the procaryotic population resulted in an absence of large structures typical of fungal mycelia or diatoms, as evidenced by scanning electron microscopy, and a significantly larger proportion of anteiso- and isobranched C15 fatty acid esters, C17 cyclopropane fatty acid esters, and the cis-vaccenic isomer of the C18 monoenoic fatty acid esters. As determined by these techniques, a marine settling community showed greater differences in bacterial as contrasted to microeucaryotic populations when compared with the microbial communities of benthic cores.  相似文献   

7.
Abstract The impact of clear-cutting, scarification, and prescribed burning on forest soil microbial community structure was assessed using sole-carbon-source utilization (SCSU). Organic and mineral soil samples were collected on two dates from Pinus banksiana plots that had been clear-cut, clear-cut followed by prescribed burning, clear-cut followed by scarification, or had not been harvested. Microorganisms were extracted from the soil samples and used to inoculate Gram-negative Biolog? plates. Patterns of substrate metabolism were used to calculate Shannon, Simpson, McIntosh, and related evenness indices. Principal component analysis (PCA) resolved organic and mineral soils. Organic soil exhibited higher metabolic diversity than mineral soil. Scarified plots showed lower diversity on one date, when diversity indices were calculated using all carbon sources, and on both dates when calculated using carboxylic acids, only. The results suggest that SCSU may be used to assess the impact of forestry practices on microbial diversity and community structure by using a subset of carbon substrates. Received: 30 July 1996; Accepted 18 November 1996  相似文献   

8.
Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about the overall structure of terrestrial microbial communities. PLFA profiling has been extensively used in a range of ecosystems as a biological index of overall soil quality, and as a quantitative indicator of soil response to land management and other environmental stressors.The standard method presented here outlines four key steps: 1. lipid extraction from soil samples with a single-phase chloroform mixture, 2. fractionation using solid phase extraction columns to isolate phospholipids from other extracted lipids, 3. methanolysis of phospholipids to produce fatty acid methyl esters (FAMEs), and 4. FAME analysis by capillary gas chromatography using a flame ionization detector (GC-FID). Two standards are used, including 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine (PC(19:0/19:0)) to assess the overall recovery of the extraction method, and methyl decanoate (MeC10:0) as an internal standard (ISTD) for the GC analysis.  相似文献   

9.
Duplicate cultures of 53 strains representing 9 species of Neisseria were analyzed by gas-liquid chromatography for cellular fatty acids. N. sicca, N. mucosa, N. flava, N. flavescens, N. perflava, N. subflava, and the several serotypes of N. meningitidis examined were found to comprise a fairly homogeneous group on the basis of the percentages of individual fatty acids present. Lactose-fermenting Neisseria also were in this group. N. catarrhalis, however, contained decanoic acid in addition to the acids occurring in the other species. Moreover, the 18 C-saturated and monoenoic acids together constituted 36% of the total fatty acid composition for N. catarrhalis, while the comparable mean value for the other species was less than 13%.  相似文献   

10.
The impact of acid rock drainage (ARD) and eutrophication on microbial communities in stream sediments above and below an abandoned mine site in the Adelaide Hills, South Australia, was quantified by PLFA analysis. Multivariate analysis of water quality parameters, including anions, soluble heavy metals, pH, and conductivity, as well as total extractable metal concentrations in sediments, produced clustering of sample sites into three distinct groups. These groups corresponded with levels of nutrient enrichment and/or concentration of pollutants associated with ARD. Total PLFA concentration, which is indicative of microbial biomass, was reduced by >70% at sites along the stream between the mine site and as far as 18 km downstream. Further downstream, however, recovery of the microbial abundance was apparent, possibly reflecting dilution effect by downstream tributaries. Total PLFA was >40% higher at, and immediately below, the mine site (0–0.1 km), compared with sites further downstream (2.5–18 km), even after accounting for differences in specific surface area of different sediment samples. The increased microbial population in the proximity of the mine source may be associated with the presence of a thriving iron-oxidizing bacteria community as a consequence of optimal conditions for these organisms while the lower microbial population further downstream corresponded with greater sediments metal concentrations. PCA of relative abundance revealed a number of PLFAs which were most influential in discriminating between ARD-polluted sites and the rest of the sites. These PLFA included the hydroxy fatty acids: 2OH12:0, 3OH12:0, 2OH16:0; the fungal marker: 18:26; the sulfate-reducing bacteria marker 10Me16:17; and the saturated fatty acids 12:0, 16:0, 18:0. Partial constrained ordination revealed that the environmental parameters with the greatest bearing on the PLFA profiles included pH, soluble aluminum, total extractable iron, and zinc. The study demonstrated the successful application of PLFA analysis to rapidly assess the toxicity of ARD-affected waters and sediments and to differentiate this response from the effects of other pollutants, such as increased nutrients and salinity.  相似文献   

11.
The whole-cell lipid extraction to profile microbial communities on soils using fatty acid (FA) biomarkers is commonly done with the two extractants associated with the phospholipid fatty acid (PLFA) or Microbial IDentification Inc. (MIDI) methods. These extractants have very different chemistry and lipid separation procedures, but often shown a similar ability to discriminate soils from various management and vegetation systems. However, the mechanism and the chemistry of the exact suite of FAs extracted by these two methods are poorly understood. Therefore, the objective was to qualitatively and quantitatively compare the MIDI and PLFA microbial profiling methods for detecting microbial community shifts due to soil type or management. Twenty-nine soil samples were collected from a wide range of soil types across Oregon and extracted FAs by each method were analyzed by gas chromatography (GC) and GC-mass spectrometry. Unlike PLFA profiles, which were highly related to microbial FAs, the overall MIDI-FA profiles were highly related to the plant-derived FAs. Plant-associated compounds were quantitatively related to particulate organic matter (POM) and qualitatively related to the standing vegetation at sampling. These FAs were negatively correlated to respiration rate normalized to POM (RespPOM), which increased in systems under more intensive management. A strong negative correlation was found between MIDI-FA to PLFA ratios and total organic carbon (TOC). When the reagents used in MIDI procedure were tested for the limited recovery of MIDI-FAs from soil with high organic matter, the recovery of MIDI-FA microbial signatures sharply decreased with increasing ratios of soil to extractant. Hence, the MIDI method should be used with great caution for interpreting changes in FA profiles due to shifts in microbial communities.  相似文献   

12.
Abstract Phospholipid fatty acid (PLFA) profiles were measured in soils from organic, low-input, and conventional farming systems that are part of the long term Sustainable Agriculture Farming Systems (SAFS) Project. The farming systems differ in whether their source of fertilizer is mineral or organic, and in whether a winter cover crop is grown. Sustained increases in microbial biomass resulting from high organic matter inputs have been observed in the organic and low-input systems. PLFA profiles were compared to ascertain whether previously observed changes in biomass were accompanied by a change in the composition of the microbial community. In addition, the relative importance of environmental variables on PLFA profiles was determined. Redundancy analysis ordination showed that PLFA profiles from organic and conventional systems were significantly different from April to July. On ordination plots, PLFA profiles from the low-input system fell between organic and conventional systems on most sample dates. A group of fatty acids (i14:0, a15:0, 16:1ω7c, 16:1ω5c, 14:0, and 18:2ω6c) was enriched in the organic plots throughout the sampling period, and another group (10Me16:0, 2OH 16:1 and 10Me17:0) was consistently lower in relative abundance in the organic system. In addition, another group (15:0, a17:0, i16:0, 17:0, and 10Me18:0) was enriched over the short term in the organic plots after compost incorporation. The relative importance of various environmental variables in governing the composition of microbial communities could be ranked in the order: soil type > time > specific farming operation (e.g., cover crop incorporation or sidedressing with mineral fertilizer) > management system > spatial variation in the field. Measures of the microbial community and soil properties (including microbial biomass carbon and nitrogen, substrate induced respiration, basal respiration, potentially mineralizable nitrogen, soil nitrate and ammonium, and soil moisture) were seldom associated with the variation in the PLFA profiles. Received: 3 February 1997; Accepted: 7 August 1997  相似文献   

13.
Although starvation survival has been characterized for many bacteria, few subsurface bacteria have been tested, and few if any have been tested in natural subsurface porous media. We hypothesized that subsurface bacteria may be uniquely adapted for long-term survival in situ. We further hypothesized that subsurface conditions (sediment type and moisture content) would influence microbial survival. We compared starvation survival capabilities of surface and subsurface strains of Pseudomonas fluorescens and a novel Arthrobacter sp. in microcosms composed of natural sediments. Bacteria were incubated for up to 64 weeks under saturated and unsaturated conditions in sterilized microcosms containing either a silty sand paleosol (buried soil) or a sandy silt nonpaleosol sediment. Direct counts, plate counts, and cell sizes were measured. Membrane phospholipid fatty acid (PLFA) profiles were quantified to determine temporal patterns of PLFA stress signatures and differences in PLFAs among strains and treatments. The Arthrobacter strains survived better than the P. fluorescens strains; however, differences in survival between surface and subsurface strains of each genus were not significant. Bacteria survived better in the paleosol than in the nonpaleosol and survived better under saturated conditions than under unsaturated conditions. Cell volumes of all strains decreased; however, sediment type and moisture did not influence rates of miniaturization. Both P. fluorescens strains showed PLFA stress signatures typical for gram-negative bacteria: increased ratios of saturated to unsaturated fatty acids, increased ratios of trans- to cis-monoenoic fatty acids, and increased ratios of cyclopropyl to monoenoic precursor fatty acids. The Arthrobacter strains showed few changes in PLFAs. Environmental conditions strongly influenced PLFA profiles.  相似文献   

14.
The effects of long-term heavy metal deposition on microbial community structure and the level of bacterial community tolerance were studied along two different gradients in Scandinavian coniferous forest soils. One was near the Harjavalta smelter in Finland, and one was at Ronnskar in Sweden. Phospholipid fatty acid (PLFA) analysis revealed a gradual change in soil microbial communities along both pollution gradients, and most of the individual PLFAs changed similarly to metal pollution at both sites. The relative quantities of the PLFAs br18:0, br17:0, i16:0, and i16:1 increased with increasing heavy metal concentration, while those of 20:4 and 18:2(omega)6, which is a predominant PLFA in many fungi, decreased. The fungal part of the microbial biomass was found to be more sensitive to heavy metals. This resulted in a decreased fungal/bacterial biomass ratio along the pollution gradient towards the smelters. The thymidine incorporation technique was used to study the heavy metal tolerance of the bacteria. The bacterial community at the Harjavalta smelter, exposed mainly to Cu deposition, exhibited an increased tolerance to Cu but not to Cd, Ni, and Zn. At the Ronnskar smelter the deposition consisting of a mixture of metals increased the bacterial community tolerance to all tested metals. Both the PLFA pattern and the bacterial community tolerance were affected at lower soil metal concentrations than were bacterial counts and bacterial activities. At Harjavalta the increased Cu tolerance of the bacteria and the change in the PLFA pattern of the microbial community were found at the same soil Cu concentrations. This indicated that the altered PLFA pattern was at least partly due to an altered, more metal-tolerant bacterial community. At Ronnskar, where the PLFA data varied more, a correlation between bacterial community tolerance and an altered PLFA pattern was found up to 10 to 15 km from the smelter. Farther away changes in the PLFA pattern could not be explained by an increased community tolerance to metals.  相似文献   

15.
Disturbances typically associated with the study of soil microbial communities, i.e., sieving, storage, and subsequent incubation at elevated temperatures, were investigated with phospholipid fatty acid (PLFA) analyses. Treatment effects were quantified by statistical analyses of the mole percentage distribution of the individual fatty acids. Changes in the concentrations of individual fatty acids over a 7-week storage period at 4.5°C were generally not statistically significant. Sieving effects (mesh size, 4 or 2 mm) on CO2 evolution and the PLFA profile were monitored over 3 weeks; the physical disturbance had only minor effects, although some damage to fungal hyphae by the first sieving (<4 mm) was suggested by a decrease in the signature fatty acid 18:2 ω6c. Temperature effects were investigated by incubating soil for up to 3 weeks at 4.5, 10, or 25°C. Principal component analyses demonstrated a significant shift in the PLFA composition at 25°C over the first 2 weeks, while changes at the other two temperatures were minor. Several of the changes observed at 25°C could be explained with reference to mechanisms of temperature adaptation or as a response to conditions of stress, including a decrease in the degree of unsaturation, an increased production of cyclopropyl fatty acids, and increased ratios of the branched-chain fatty acids iso-15:0 and iso-17:0 over anteiso-15:0 and anteiso-17:0, respectively. A decrease in the total amount of PLFA was also indicated.  相似文献   

16.
用磷脂脂肪酸法(PLFA)分析广东鹤山两种人工林——针叶林和荷木林不同土层(0~20 cm、20~40 cm、40~60 cm)的土壤微生物群落结构。结果表明,两种人工林的磷脂脂肪酸总量(PLFAs总量)、细菌特征脂肪酸(细菌PLFAs)、真菌特征脂肪酸(真菌PLFAs),以及革兰氏阳性菌和革兰氏阴性菌特征脂肪酸的含量都是0~20 cm土层最高;荷木林的PLFAs总量高于针叶林,但这种差异随土壤深度增加而递减, 且与土壤有机碳、全氮含量呈正相关。在相同土层,细菌PLFAs含量均显著高于真菌PLFAs含量。主成分分析表明,土壤微生物多样性及其丰富度受林型和土层综合作用的影响;单一的PLFAs中,i13:0、a13:0、17:0、cy17:0和16:0等脂肪酸对第一主成分的贡献较大;i16:0、cy19:0、18:1ω9c、i15:0、18:2ω6c和a17:0等脂肪酸对第二主成分贡献较大。  相似文献   

17.
The fatty acid composition of total lipids in two species of sponges (Halichondria panicea and Clathria pennata) inhabiting the same biotope in the Sea of Japan was examined. More than 80 fatty acids (FAs) were found for each species of sponges. Of them, 61 fatty acids were identified for H. panicea and 54 for C. pennata. The relative content of most FAs was less than 1%. The contribution of symbiotic microorganisms to the total fatty acids was higher in H. panicea than in C. pennata. Bacterial symbionts and microeukaryotes were found among the microorganisms associated with the sponges. The contribution of prokaryotic organisms was equal (7.8%) for both species of sponges, but the fatty acids were characteristic of different taxonomic groups of bacteria. The proportion of microeukaryotic fatty acids in the total lipids of H. panicea (19.6%) was two times that of C. pennata (10.1%).  相似文献   

18.
Abstract The patterns of seasonal variation in the structure of a marine benthic microbial community were examined using phospholipid fatty acid analysis (PLFA). Principal component analysis of PLFA profiles indicated a strong seasonal pattern dominated the variance within the data set. Three functional groups of microorganisms (phototrophic microeukaryotes, and two groups of anaerobic bacteria) were disproportionately abundant in the communities that mapped to either extreme of the first principle component. Phototrophic microeukaryotes were most abundant and exhibited the greatest relative abundance during periods of cold water. In contrast, the two functional groups of anaerobic bacteria showed the greatest relative abundance during times of warm water. Differential responses by these groups, and macrofaunal deposit feeders, to light intensity and water temperature were offered as the proximal causes of the observed patterns. Received: 28 April 1997; Accepted: 10 September 1997  相似文献   

19.
Ester-linked phospholipid fatty acid (PLFA) profiles of a Pseudomonas aureofaciens strain and an Arthrobacter protophormiae strain, each isolated from a subsurface sediment, were quantified in a starvation experiment in a silica sand porous medium under moist and dry conditions. Washed cells were added to sand microcosms and maintained under saturated conditions or subjected to desiccation by slow drying over a period of 16 days to final water potentials of approximately - 7.5 MPa for the P. aureofaciens and - 15 MPa for the A. protophormiae. In a third treatment, cells were added to saturated microcosms along with organic nutrients and maintained under saturated conditions. The numbers of culturable cells of both bacterial strains declined to below detection level within 16 days in both the moist and dried nutrient-deprived conditions, while direct counts and total PLFAs remained relatively constant. Both strains of bacteria maintained culturability in the nutrient-amended microcosms. The dried P. aureofaciens cells showed changes in PLFA profiles that are typically associated with stressed gram-negative cells, i.e., increased ratios of saturated to unsaturated fatty acids, increased ratios of trans- to cis-monoenoic fatty acids, and increased ratios of cyclopropyl fatty acids to their monoenoic precursors. P. aureofaciens starved under moist conditions showed few changes in PLFA profiles during the 16-day incubation, whereas cells incubated in the presence of nutrients showed decreases in the ratios of both saturated fatty acids to unsaturated fatty acids and cyclopropyl fatty acids to their monoenoic precursors. The PLFA profiles of A. protophormiae changed very little in response to either nutrient deprivation or desiccation. Diglyceride fatty acids, which have been proposed to be indicators of dead or lysed cells, remained relatively constant throughout the experiment. Only the A. protophormiae desiccated for 16 days showed an increase in the ratio of diglyceride fatty acids to PLFAs. The results of this laboratory experiment can be useful for interpreting PLFA profiles of subsurface communities of microorganisms for the purpose of determining their physiological status.  相似文献   

20.
简要介绍微生物产多不饱和脂肪酸的生化研究现状,着重从高产菌株的筛选、工程菌株的构建、发酵条件及产业化现状等方面论述微生物发酵生产多不饱和脂肪酸的主要研究进展;概述多不饱和脂肪酸的提取制备技术,并对发酵法生产多不饱和脂肪酸研究目标和发展前景提出了建议.  相似文献   

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