Phospholipid fatty acid and infra-red spectroscopic analysis of a sulphate-reducing consortium

Abstract In order to validate unusual fatty acids as biomarkers for sulphate-reducing bacteria, selective conditions were arranged for the enrichment of a marine glutamate-fermenting bacterium which made hydrogen and acetate available for oxidation via the respiration of sulphate. Under these conditions the complete oxidation of glutamate via sulphate reduction accounted for 84% of the available electron equivalents. Fatty acid biomarkers for hydrogen-oxidizing Desulfovibrio sp. (iso 17:1w7c and branched monoenoics) and for acetate-oxidizing Desulfobacter (10 methyl 16:0) were detected in the enrichment. These biomarkers were demonstrated in pure cultures of Desulfovibrio sp. and Desulfobacter sp. obtained from the enrichment. The predominant glutamate-fermenting bacterium isolated from the consortium contained no branched ester-linked phospholipid fatty acids, and produced acetate and hydrogen. With energy limitation the enriched consortium produced increased amounts of extracellular polysaccharide and the endogenous storage lipid poly-beta-hydroxybutyrate as detected with Fourier transform/infra-red (FT-IR) spectroscopy.     

Signature fatty acids in the polar lipids of acid-producing Thiobacillus spp.: Methoxy, cyclopropyl, alpha-hydroxy-cyclopropyl and branched and normal monoenoic fatty acids

Abstract The polar lipids of 5 species of Thiobacillus were extracted and purified. An analysis of the fatty acid composition of the polar lipids documented the presence of methoxy, cyclopropyl, monounsaturated and hydroxycyclopropyl fatty acids of sufficiently unusual structure to serve as 'signatures' for the presence of these organisms in environmental samples. The structures of the unusual fatty acids of the polar lipids were confirmed by mass spectrometry (MS) after isolation by capillary gas chromatography (GC).     

Polar lipid biomarkers of free-living bacteria from oligotrophic marine waters

Free-living marine bacteria isolated from oligotrophic Mediterranean waters were enriched in culture to characterize their phospholipid fatty acids (PLFAs). Odd chain iso- and anteiso-FAMEs and n–16:0 were the predominant structural PLFAs, together with a homologous series identified as mid-chain methoxy FAMEs. The dominant methoxy fatty acids identified were 9-CH₃O-15:0, 9-CH₃-16:0 and 11-CH₃O-17:0, occuring as pairs of stereoimers. Methoxy fatty acids accounted for up to 37% of PLFAs of free-living bacteria, which sets them as promising new biomarkers for bacteria of oligotrophic waters. Although similar homologues have already been characterized in a variety of eukaryotes and prokaryotes, methoxy fatty acids are identified here for the first time in marine bacteria. Analytical difficulties that may hinder the characterization of these biomarkers are presented, and structural elucidation keys by gas chromotography coupled to mass spectrometry are discussed. Whilst bacterial branched fatty acids were transferred to storage lipids of bacterivorous flagellates methoxy acids were not transferred to higher trophic levels in the studied conditions.     

Lipid content and related fatty acid composition in fallow and cultivated soil in Southern Italy

Summary Lipid content and fatty acid composition were studied on two soil samples—one fallow and one cultivated with durum wheat. Differences between the cultivated and the fallow subunits in respect of total lipid content and the saponifiable fraction were found to be statistically significant. The same observations were made on some fatty acids showing either accumulation or decrease after three years. The interaction years x cultivation for total lipid content and the saponifiable fraction was significant showing that accumulation and reduction of lipids in the cultivated subunit are in equilibrium. The interaction dates of sampling x cultivation was significant for C150, C160, C180, C240 as their contents increase in the cultivated subunit in July as a result of the decomposition of plant tissues.Results discussed in this paper have been partially presented at the 7th International Symposium Humus et Planta Brno (Czechoslovakia) on August 20–25th 1979. This research has been partially supported by the National Research Council of Italy and carried out at the Experimental Institute of Agronomy — Bari — Italy (Director: F. Lanza).     

Low pH-induced membrane fatty acid alterations in oral bacteria

Four oral bacterial strains, of which two are considered aciduric and two are considered acid-sensitive, were grown under glucose-limiting conditions in chemostats to determine whether their membrane fatty acid profiles were altered in response to environmental acidification. Streptococcus gordonii DL1, as well as the aciduric strains S. salivarius 57.I, and Lactobacillus casei 4646 increased the levels of mono-unsaturated membrane fatty acids. The non-aciduric strain S. sanguis 10904 did not alter its membrane composition in response to pH values examined here. Thus, in response to low pH, aciduric oral bacteria alter their membrane composition to contain increased levels of long-chained, mono-unsaturated fatty acids. This suggests that membrane fatty acid adaptation is a common mechanism utilized by bacteria to withstand environmental stress.     

Lipid and fatty acid biomarkers as proxies for environmental contamination in caged mussels Mytilus galloprovincialis

Mussels (Mytilus galloprovincialis) were transplanted from a reference site (Syracuse harbour) to an impacted site (Augusta Bay) from January to July 2013 to assess the biochemical response of caged mussels to high trace element and polycyclic aromatic hydrocarbon (PAH) contamination, using lipid and fatty acid (FA) biomarkers. Sediment and mussels were analysed to assess contaminant bioavailability in the study sites and bioaccumulation in mussel tissue. Trace elements (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, V, Zn) and PAHs were significantly higher in mussels from Augusta than in those from Syracuse, mirroring the different environmental contamination. The biological impact quotient (BIQs), which indicates the potential biological impact of mussel-accumulated contaminants, reflected the highest contaminant concentrations found in Augusta and their temporal trend, which increased from the start of the experiment to 3–4 months after transplanting, followed by a decrease, as indication of mussel detoxification processes. Lower condition index (CI) and phospholipids (PL), as well as higher total (TL) and neutral lipids (NL) in mussels from Augusta, indicated the occurrence of a physiological and biochemical stress response to pollutant exposure and accumulation. Differences in FA composition, especially polyunsaturated (PUFA), essential (EFA) and non-methylene interrupted dienoic (NMID) FAs between the study sites revealed the occurrence of stress-induced lipid peroxidation, followed by compensatory/adaptive processes in Augusta mussels. The marked increase in bacterial biomarkers, mainly cyclopropyl (CY) FAs, reflecting the greater bioaccumulation of chemicals in the Augusta mussels, may be the result of bacterial infections or symbiosis with bacteria involved in detoxification processes.     

Phospholipid fatty acid and lipopolysaccharide fatty acid signature lipids in methane-utilizing bacteria

The effect of human bile juice and bile salts (sodium cholate, sodium taurocholate, sodium glycochenodeoxycholate and sodium chenodeoxycholate) on growth, sporulation and enterotoxin production by enterotoxin-positive and enterotoxin-negative strains of Clostridium perfringens was determined. Each bile salt inhibited growth to a different degree. A mixture of bile salts completely inhibited the growth of enterotoxin-positive strains of this organism. Human bile juice completely inhibited the growth of all the strains at a dilution of 1:320. A distinct stimulatory effect of the bile salts on sporulation was observed in the case of C. perfringens strains NCTC 8239 and NCTC 8679. The salts also increased enterotoxin concentrations in the cell extracts of the enterotoxin-positive strains tested. No effect on enterotoxin production was detected when an enterotoxin-negative strain was examined.     

Application of high-performance liquid chromatography of plasma fatty acids as their phenacyl esters to evaluate splanchnic and renal fatty acid balance in vivo

Plasma fatty acids from renal and hepatic veins, and arterialized hand vein obtained in 20 subjects before and after insulin infusion were separated by reversed-phase high-performance liquid chromatography following phenacyl esterification. Separation and quantification over the range 1.0–100 nmol per injection of nine fatty acids was achieved within 60 min using [²H₃₁]palmitic acid as internal standard. Analytical recoveries were greater than 90% and the intra- and inter-assay coefficients of variation were less than 2.5 and 4.0%, respectively. Following insulin infusion, net splanchnic uptake of total fatty acids decreased from 3.0±0.3 to 1.0±0.1 μmol/kg min (p<0.01), whereas net renal balance remained neutral (−0.04±0.04 vs. −0.06±0.03 μmol/kg min, p=N.S.). Individual fatty acid balance varied from a low of 0.012±0.005 (myristic acid) to a high of 0.95±0.08 (oleic acid) μmol/kg min across the splanchnic tissues and from 0.005±0.002 (stearic acid) to 0.21±0.1 (oleic acid) μmol/kg min across the kidney. There is a substantial diversity in changes in plasma concentration and regional balance of individual fatty acid during short-term fasting and hyperinsulinemia. This method is simple, accurate, and can be applied to assess individual fatty acid metabolism in vivo.     

Fatty acid,menaquinone and polar lipid composition of Rothia dentocariosa

The lipid compositions of Rothia dentocariosa was investigated. All of the strains tested possessed closely related lipid profiles consisting of predominantly straight-chain saturated and methyl branched long-chain fatty acids, unsaturated menaquinones with seven isoprene units and a polar lipid composition comprising diphosphatidylglycerol, phosphatidylglycerol and a diglycosyldiacylglycerol. The results of the present study indicate Rothia dentocariosa is a good and distinct taxon. The lipid data however does not support the classification of Rothia dentocariosa in the family Actinomycetaceae.     

Selective screening for fatty acid oxidation disorders by tandem mass spectrometry: difficulties in practical discrimination

In a selective screening for fatty acid oxidation disorders by tandem mass spectrometry, we tested the diagnostic ratios and acylcarnitine concentrations in sera or blood spots, which were reported to be specific to very long-chain acyl CoA dehydrogenase deficiency, carnitine palmitoyltransferase I deficiency, and carnitine palmitoyltransferase II deficiency. While the acylcarnitine profiles in the majority of these patients were typical in the respective disorders, some overlapping of the indices was observed between these patients and the infants, who showed symptoms mainly related to hypoglycemia but did not have the disorders mentioned above. Although the diagnostic ratio of tetradecenoylcarnitine to dodecanoylcarnitine for very long-chain acyl CoA dehydrogenase deficiency seemed to minimize the overlapping in this study, additional measures including careful assessment of clinical data and enzyme assays may be necessary for the diagnosis in atypical cases.     

Ladderane lipid distribution in four genera of anammox bacteria

Intact ladderane phospholipids and core lipids were studied in four species of anaerobic ammonium oxidizing (anammox) bacteria, each representing one of the four known genera. Each species of anammox bacteria contained C(18) and C(20) ladderane fatty acids with either 3 or 5 linearly condensed cyclobutane rings and a ladderane monoether containing a C(20) alkyl moiety with 3 cyclobutane rings. The presence of ladderane lipids in all four anammox species is consistent with their putative physiological role to provide a dense membrane around the anammoxosome, the postulated site of anammox catabolism. In contrast to the core lipids, large variations were observed in the distribution of ladderane phospholipids, i.e. different combinations of hydrophobic tail (ladderane, straight chain and methyl branched fatty acid) types attached to the glycerol backbone sn-1 position, in combination with different types of polar headgroup (phosphocholine, phosphoethanolamine or phosphoglycerol) attached to the sn-3 position. Intact ladderane lipids made up a high percentage of the lipid content in the cells of "Candidatus Kuenenia stuttgartiensis", suggesting that ladderane lipids are also present in membranes other than the anammoxosome. Finally, all four investigated species contained a C(27) hopanoid ketone and bacteriohopanetetrol, which, indicates that hopanoids are anaerobically synthesised by anammox bacteria.     

Differences in lipid and in fatty acid composition of taproots and lateral roots of faba bean plants (Vicia faba L.) grown in saline media

Abstract

The content and composition of lipids and fatty acids of taproots and lateral roots of Vicia faba were investigated with plants grown under saline (80 mM NaCl) and non-saline conditions. Lipids of both types of faba bean roots are constituted of ～80% phospholipids, of ～15% glycolipids and of some 5% of wax-esters. Phosphatidylethanolamine (PE) and phosphatidylcholine are the main constituents of both root types. Di-phosphatyidylglycerol and phosphatidylinositol are present in medium concentrations, whereas phosphatidylglycerol and phosphatidylserine are present only in trace amounts. The content of sulpholipids was much lower in salt-treated roots as compared with that of the controls. The content of cardiolipins and PE was higher in lateral roots than in the taproots. Roots of salt-treated plants had some 18% lower ether-soluble lipid content in the lateral roots, and approximately 28% lower than the taproots. Less than 25% of the fatty acids of the extracted phospholipids were saturated, with palmitic acid as the main constituent (13 – 18%). Linoleic acid comprised 65 – 70% of the unsaturated acids. Differences in the composition of some fatty acids were found between taproots and lateral roots of salt-treated plants. The observed differences in composition between root types suggests that the reported physiological differences between such roots could be based, at least in part, on structural or compositional differences in their lipids.     

Comparative study of the fatty acid composition of some groups of purple nonsulfur bacteria

The fatty acid composition (FAC) of 43 strains of purple nonsulfur bacteria belonging to six genera—Rubrivivax, Rhodopseudomonas, Rhodoplanes, Blastochloris, Rhodobium, and Rhodomicrobium—was studied by capillary gas chromatography. The cultures were grown on standard medium under standard conditions. Automatic identification of the fatty acid methyl esters and statistical processing of the results were performed by the computerized Microbial Identification System (MIS). Significant differences between the FACs of different genera, species, and, sometimes, strains were revealed. 16S rRNA genes of some of the new isolates, primarily those having a specific FAC, were sequenced. The taxonomic status of a number of the strains in question was determined using the FAC characteristics as one of the criteria. It was shown that the FAC characteristics may be used both for affiliating isolates to known species and for revealing new taxa.     

The effect of unsaturated and saturated dietary lipids on the pattern of daily torpor and the fatty acid composition of tissues and membranes of the deer mouse Peromyscus maniculatus

Summary Dieary lipids strongly influence the pattern of torpor and the body lipid composition of mammalian hibernators. The object of the present study was to investigate whether these diet-induced physiological and biochemical changes also occur in species that show shallow, daily torpor. Deer mice, Peromyscus maniculatus, were fed with rodent chow (control diet) or rodent chow with either 10% sunflower seed oil (unsaturated diet) or 10% sheep fat (saturated diet). Animals on the unsaturated diet showed a greater occurrence of torpor (80–100% vs 26–43%), longer torpor bouts (4.5 vs 2.25 h), a lower metabolic rate during torpor (0.96 vs 2.25 ml O₂·g^-1·h^-1), and a smaller loss of body mass during withdrawal of food (2.35 vs 3.90 g) than animals on the saturated diet; controls were intermediate. These diet-induced physiological changes were associated with significant alterations in the fatty acid composition of depot fat, leg muscle and brain total lipids, and heart mitochondrial phospholipids. Significant differences in the total unsaturated fatty acid (UFA) content between animals on saturated and unsaturated diet were observed in depot fat (55.7% vs 81.1%) and leg muscle (56.4% vs 72.1%). Major compositional differences between diet groups also occurred in the concentration of n6 and/or n3 fatty acids of brain and heart mitochondria. The study suggests that dietary lipids may play an important role in the seasonal adjustment of physiology in heterothermic mammals.Abbreviations EDTA ethylenediaminetetra-acetic acid - HEPES N-2 hydroxyethylpiperazine-N¹-2-ethanesulphonic acid - MUFA monounsaturated fatty acids - PUFA polyunsaturated fatty acids - RMR Testing metabolic rate - SD standard deviation - SFA saturated fatty acids - SNK Student-Newman-Keuls test - T₁ air temperature - T_b body temperature - UFA unsaturated fatty acids - rate of oxygen consumption Dedicated to the late John K. Raison     

Comparative study of fatty acid composition in human and monkey aorta

The fatty acid composition of thoracic and abdominal aortic intima and media of normal human subjects and rhesus monkeys has been studied. Significantly higher values of unsaturated fatty acids as compared to saturated fatty acids have been noted in the intima of monkey as compared to man. The fatty acid profile of the aortic wall in these two species has provided a probable biochemical basis for the lesser incidence of atherosclerosis in macaques.     

Membrane fatty acid analysis of Antarctic bacteria

Abstract Randomly selected strains of a bacterial collection of marine sea-ice bacteria from Antarctica were analyzed to obtain a profile of the membrane fatty acids. Results showed that short chain saturated and unsaturated fatty acids were more common in the psychrotrophs when compared to psychrophiles. In contrast, branched-chain fatty acids were more abundant in the psychrophiles.     

Effects of drought on [1-14C]-oleic and [1-14C]-linoleic acid desaturation in cotton leaves

The effects of water stress on [1-¹⁴C]-oleic and [1-¹⁴C]-linoleic acid desaturations were studied in leaves of two varieties of cotton ( Gossypium hirsutum L.), one drought-sensitive (Reba) and the other more resistant (Mocosinho). After 24 h incorporation, [1-¹⁴C]-oleate led to the appearance of linoleate in phospholipids and, additionally, of linolenate in galactolipids. [1-¹⁴C]-Linoleate was desaturated to linolenate only in galactolipid fractions. Water stress markedly inhibited the incorporation of the precursors into the leaf lipids. The two desaturation steps were affected, particularly the transformation of linoleate to linolenate in monogalactosyldiacylglycerol in the drought-sensitive variety of cotton. The metabolic implications of the inhibition of the biosynthesis of C₁₈-polyunsaturated fatty acids are discussed.     

Population study of Astarte sulcata, da Costa, 1778, (Mollusca,Bivalvia) from two Norwegian fjords based on the fatty acid composition of the adductor muscle

Multivariate analyses of fatty acid composition of Astarte sulcata, collected in Northwest Norway in October 2005, showed significant differences between geographically close populations. This biochemical heterogeneity may have been caused by high genetic drift due to the direct development of the species. The results indicate that deep basins may provide an alternative bathymetric guide for spreading of offspring rather than being carried on currents crossing such basins. Thus, the combination of biology and topography might have led to the observed pattern that populations from opposite sides of a fjord are biochemically more distant than populations along the same side of a fjord/basin.     

Effects of legacy nuclear waste on the compositional diversity and distributions of sulfate-reducing bacteria in a terrestrial subsurface aquifer

The impact of legacy nuclear waste on the compositional diversity and distribution of sulfate-reducing bacteria in a heavily contaminated subsurface aquifer was examined. dsrAB clone libraries were constructed and restriction fragment length polymorphism (RFLP) analysis used to evaluate genetic variation between sampling wells. Principal component analysis identified nickel, nitrate, technetium, and organic carbon as the primary variables contributing to well-to-well geochemical variability, although comparative sequence analysis showed the sulfate-reducing bacteria community structure to be consistent throughout contaminated and uncontaminated regions of the aquifer. Only 3% of recovered dsrAB gene sequences showed apparent membership to the Deltaproteobacteria. The remainder of recovered sequences may represent novel, deep-branching lineages that, to our knowledge, do not presently contain any cultivated members; although corresponding phylotypes have recently been reported from several different marine ecosystems. These findings imply resiliency and adaptability of sulfate-reducing bacteria to extremes in environmental conditions, although the possibility for horizontal transfer of dsrAB is also discussed.     

Growth, natural relationships, cellular fatty acids and metabolic adaptation of sulfate-reducing bacteria that utilize long-chain alkanes under anoxic conditions

Natural relationships, improvement of anaerobic growth on hydrocarbons, and properties that may provide clues to an understanding of oxygen-independent alkane metabolism were studied with two mesophilic sulfate-reducing bacteria, strains Hxd3 and Pnd3. Strain Hxd3 had been formerly isolated from an oil tank; strain Pnd3 was isolated from marine sediment. Strains Hxd3 and Pnd3 grew under strictly anoxic conditions on n-alkanes in the range of C₁₂–C₂₀ and C₁₄–C₁₇, respectively, reducing sulfate to sulfide. Both strains shared 90% 16 S rRNA sequence similarity and clustered with classified species of completely oxidizing, sulfate-reducing bacteria within the δ-subclass of Proteobacteria. Anaerobic growth on alkanes was stimulated by α-cyclodextrin, which served as a non-degradable carrier for the hydrophobic substrate. Cells of strain Hxd3 grown on hydrocarbons and α-cyclodextrin were used to study the composition of cellular fatty acids and in vivo activities. When strain Hxd3 was grown on hexadecane (C₁₆H₃₄), cellular fatty acids with C-odd chains were dominant. Vice versa, cultures grown on heptadecane (C₁₇H₃₆) contained mainly fatty acids with C-even chains. In contrast, during growth on 1-alkenes or fatty acids, a C-even substrate yielded C-even fatty acids, and a C-odd substrate yielded C-odd fatty acids. These results suggest that anaerobic degradation of alkanes by strain Hxd3 does not occur via a desaturation to the corresponding 1-alkenes, a hypothetical reaction formerly discussed in the literature. Rather an alteration of the carbon chain by a C-odd carbon unit is likely to occur during activation; one hypothetical reaction is a terminal addition of a C₁ unit. In contrast, fatty acid analyses of strain Pnd3 after growth on alkanes did not indicate an alteration of the carbon chain by a C-odd carbon unit, suggesting that the initial reaction differed from that in strain Hxd3. When hexadecane-grown cells of strain Hxd3 were resuspended in medium with 1-hexadecene, an adaptation period of 2 days was observed. Also this result is not in favor of an anaerobic alkane degradation via the corresponding 1-alkene. Received: 25 June 1998 / Accepted: 29 July 1998