Biomedical subjects

P D Nichols

Publications and source records attributed to P D Nichols.

At least 19 recordsLinked to original sources

PubMed2001

Interannual variations in the lipids of the Antarctic pteropods Clione limacina and Clio pyramidata.

Antarctic pteropods, Clione limacina (Order Gymnosomata) and Clio pyramidata (order Thecosomata), were collected near Elephant Island, South Shetland Islands, during 1997 and 1998. Total lipid was high in C. limacina (29--36 mg g(-1) wet mass) and included 46% of diacy1glyceryl ether (DAGE, as % of total lipid) for both 1997 and 1998. DAGE was not detected in C. pyramidata, which had mainly polar lipid and triacy1glycerol. 1-O-Alkyl glyceryl ethers (GE) derived from the DAGE consisted primarily of 15:0 and 16:0, with lower 17:0 and a17:0. The principal sterols of both pteropods included trans-dehydrocholesterol, brassicasterol, 24-methylenecholesterol, cholesterol and desmosterol. Levels of 24-methylenecholesterol and desmosterol were lower in both pteropods in 1997 compared to 1998. C. limacina had high levels of the odd-chain fatty acids 17:1(n--8)c and 15:0 in contrast to C. pyramidata. The previously proposed source of elevated odd-chain fatty acids in C. limacina is via propionate derived from phytoplankton DMPT; another possible source may be from thraustochytrids, which are common marine microheterotrophs. C. pyramidata had twice as much PUFA as C. limacina, largely due to higher 20:5(n--3). The PUFA 18:5(n--3) and very long chain fatty acids (C(24), C(26) and C(28) VLC-PUFA) were only detected in 1998 pteropods. In comparison, 1996 samples of C. limacina contained lower DAGE levels, which also may reflect differences in diet and oceanographic conditions. Interannual variations in specific lipid biomarkers are discussed with respect to possible different phytoplankton food sources available in the AMLR survey area.

Animals↗

PubMed2000

Evaluation of extraction methods for recovery of fatty acids from lipid-producing microheterotrophs.

The effect of different extraction techniques on the recovery of fatty acids from freeze-dried biomass of two lipid-producing microheterotrophs was examined. Two procedures were used: the extraction of lipids from biomass followed by transesterification of the fatty acids (extraction-transesterification); and the direct transesterification of biomass to produce fatty acid methyl esters (i.e. without the initial extraction step). Variable factors in the extraction-transesterification experiment were the sequence in which solvents were added to the samples, the relative amount of methanol in the solvent mix, and sonication of biomass while in the solvent mix. Variable factors in the direct transesterification experiment were sample size, and reaction duration. Statistical analysis of data (level of significance P<0.05) showed that: (1) extraction of total fatty acids prior to transesterification was significantly more efficient when solvents were added in the order of increasing polarity; (2) neither sonication nor increasing the proportion of methanol in the extraction solvent significantly affected extraction of fatty acids prior to transesterification; (3) efficiency of direct transesterification of fatty acids increased significantly with reaction time; (4) efficiency of direct transesterification of fatty acids was not significantly affected by sample size; (5) the most efficient method for extraction of fatty acids prior to transesterification yielded significantly less fatty acids than the most effective direct transesterification method. While the study examined only two strains, our results suggest that fatty acid analysis methodology for microheterotrophs under consideration for biotechnological exploitation requires optimisation and validation.

Biomass↗

PubMed2000

Lipids of gelatinous Antarctic zooplankton: Cnidaria and Ctenophora.

Cnidaria (Calycopsis borchgrevinki, Diphyes antarctica, Stygiomedusa gigantea, Atolla wyvillei, Dimophyes arctica) and Ctenophora (Beroe cucumis, B. forskalii, Pleurobrachia pileus, Bolinopsis infundibulum) were collected near Elephant Island, South Shetland Islands, during January and February 1997 and 1998. Total lipid was low in all zooplankton (0.1-5 mg g wet mass) and included primarily polar lipids (59-96% of total lipid). Triacylglycerols were 0-26% of total lipids, and wax esters were 0-11% in all species. Cholesterol was the major sterol in all Cnidaria (50-63% of total sterols) whereas in most ctenophores it was lower at 26-45%. These cholesterol levels are consistent with a combined carnivorous and phytoplanktivorous diet in the ctenophores, with the carnivorous diet more dominant in the Cnidaria. Other sterols included primarily trans-dehydrocholesterol, desmosterol, 24-methylcholest-5,22E-dien-3beta-ol, 24-nordehydrocholesterol, and 24-methylenecholesterol. Total stanols were 0-6% in all zooplankton. Eicosapentaenoic acid and docosahexaenoic acid were the major polyunsaturated fatty acids (PUFA) in all samples (7-25% of total fatty acids) except for A. wyvillei in which docosapentaenoic acid was 10% of total fatty acids. The PUFA 18:5n-3 was not detected in 1997 samples, but constituted 0.2-0.8% in most 1998 samples. Monounsaturated fatty acids included primarily 18:1n-9c, 16:1n-7c, and 18:1n-7c. The principal saturated fatty acids in all samples were 16:0, 18:0, and 14:0. These data are the first for many of these zooplankton species and the first sterol data for most species. The use of the signature lipid approach has enabled examination of aspects of trophodynamics not obtainable by conventional techniques.

Animals↗

PubMed2000

Lipid composition of the liver oil of deep-sea sharks from the Chatham Rise, New Zealand.

Deep-sea sharks approach neutral buoyancy by means of a large liver that contains large amounts of low-density lipids, primarily squalene and diacyl glyceryl ether (DAGE). As an animal increases in size and matures sexually, many biochemical changes take place within the animal. It was hypothesized that maintenance of neutral buoyancy in deep-sea sharks involves fine-scale changes in the chemical composition of the liver oil as individual sharks grow and develop. To test this hypothesis, the lipid composition of liver oil for individuals of different size and sex of deep-sea sharks from the Chatham Rise, New Zealand was compared. The composition of liver oil varied within and among species. Several species contained large amounts of squalene and DAGE, whereas only traces of these lipids were present in other species. The amounts of squalene and DAGE in liver oil were inversely related, and squalene content tended to decrease as sharks increased in size. Species with high squalene levels (> 80%) in liver oil were not abundant on the Chatham Rise, although levels of DAGE (a lipid of increasing commercial interest) were elevated in many species. Maintenance of neutral buoyancy in deep-sea sharks appears to involve changes in the composition of low-density liver lipids as the sharks increase in size and mature.

Animals↗

PubMed1999

The occurrence and possible significance of diacylglyceryl ether lipids in abalone.

Gonadal and foot tissues of the green abalone, Haliotis fulgens, farm-raised on macroalgal [corrected] diets, were analyzed for lipids using thin-layer chromatography/flame-ionization detection and gas chromatography-mass spectrometry (GC-MS). Diacylglyceryl ether (DAGE) was 0.7% of total lipids in the gonad. The major alkyl constituents of the glyceryl ether diols in the gonad (as % of total diols) were 16:0 (38%) and 18:1 (36%). While levels of DAGE in the abalone foot were below flame-ionization detection limits, glyceryl ether diols from them were detected using the more sensitive GC-MS procedure. The major diol components in the foot were 18:0 (39%) and 18:1 (32%). To our knowledge, this is the first report of DAGE in abalone tissues. Although the precise role of DAGE in abalone remains to be determined, a possible structural role may exist.

Animals↗

PubMed1998

Flavobacterium hibernum sp. nov., a lactose-utilizing bacterium from a freshwater Antarctic lake.

Four freshwater Antarctic lakes were examined for the presence of beta-galactosidase-producing bacteria using mineral medium enrichments and lactose. Enrichments from only one of the lakes produced growth and two strains were isolated that were very similar in phenotype and fatty acid profile, and shared considerable homology in their DNA (DNA-DNA hybridization = 93 +/- 7%). The strains were psychrotrophic with theoretical Tmax, Tmin and Topt of 30-31, -7 degrees and 26 degrees C, respectively. The beta-galactosidase in cell extracts had an optimal activity at 39 degrees C. The strains were Gram-negative rods, showed gliding motility, contained branched and hydroxy fatty acids, and menaquinone 6 as the major respiratory quinone. The strains did not form microcysts and utilized lactose while using ammonium ions as a source of nitrogen, and a range of other sugars. The G + C content of the DNA was 34 mol%. Phylogenetic analysis of one of the strains, by comparison of 16S rDNA sequences, showed that it was most similar, but not identical to, Flavobacterium columnare and '[Sporocytophaga] cauliformis'. Both species could be differentiated phenotypically from the Antarctic isolates. DNA-DNA hybridization of the Antarctic isolate with six different members of the Flavobacterium 16S rDNA cluster showed no strain with greater than 18% relatedness. The nearest type species to the Antarctic isolate in the phylogenetic analysis was Flavobacterium aquatile. The name Flavobacterium hibernum is proposed for the Antarctic strains, and the type strain is ATCC 51468T (= ACAM 376T).

Antarctic Regions↗

PubMed1997

Polyunsaturated fatty acids in the psychrophilic bacterium Shewanella gelidimarina ACAM 456T: molecular species analysis of major phospholipids and biosynthesis of eicosapentaenoic acid.

The production of eicosapentaenoic acid [20:5omega3; EPA] from Shewanella gelidimarina (ACAM 456T) was investigated with respect to growth temperature and growth on sole carbon sources. The percentage and quantitative yield of EPA remained relatively constant at all growth temperatures within or below the optimal growth temperature region. At higher growth temperatures, these values decreased greatly. Growth on differing sole carbon sources also influenced the percentage and amount of EPA produced, with the fatty acid composition influenced by provision of potential acyl chain primers as sole carbon sources. The highest amounts of EPA occurred from growth on propionic acid and L-leucine respectively, while the highest percentage of EPA occurred from growth on L-proline. Monounsaturated fatty acid components and EPA were concentrated in phosphatidylglycerol (PG), while the proportion of branched-chain fatty acids was elevated in phosphatidylethanolamine (PE); the two major phospholipid classes. Specific associations of EPA with other acyl chains were identified within cellular phospholipid classes. The association of EPA with 17:1 and 18:0 acyl chains in phospholipid species was specific to PG, whereas the association of EPA with i13:0/13:0 and 14:0/i14:0 was specific to PE. Such acyl chain 'tailoring' is indicative of the important role of EPA in bacterial membrane adaptive responses. EPA was also a large component (22%) of a non-esterified fatty acid (NEFA) fraction within the total lipid extract of the bacterium. This may point toward a particular role of NEFA in polyunsaturated fatty acid (PUFA) metabolism. The formation of EPA was investigated by labelling with L-[U-14C]serine and sodium [1-14C]acetate. The accumulation of radiolabel within unsaturated intermediates (di-, tri- and tetraunsaturated fractions) was low, indicating a rapid formation and derivatisation of these components. Similar results were found for the unsaturated fatty acid fractions of both PE and PG using sodium [1-14C]acetate radiolabel. The regulation of triunsaturated fatty acid components may be a potential control site in PUFA biosynthesis.

Carbon Radioisotopes↗

PubMed1997

Acid habituation of Escherichia coli and the potential role of cyclopropane fatty acids in low pH tolerance.

A reversible adaptive tolerance to low pH termed 'acid habituation' is demonstrated for five strains of Escherichia coli. Superimposed upon the intrinsic acid tolerance of individual strains, acid habituation significantly enhances the survival of exponential phase cultures exposed to a lethal acid challenge (pH 3.0), and minimises inter-strain variability in acid tolerance. The fatty acid composition of acid habituated, non-habituated, and de-habituated exponential phase cultures is also reported. During acid habituation, monounsaturated fatty acids (16:1 omega 7c and 18:1 omega 7c) present in the phospholipids of E. coli are either converted to their cyclopropane derivatives (cy17:0 and cy19:0), or replaced by saturated fatty acids. The acid tolerance of individual strains of E. coli appears to be correlated with membrane cyclopropane fatty acid content and, thus, it is postulated that increased levels of cyclopropane fatty acids may enhance the survival of microbial cells exposed to low pH. The results presented illustrate the remarkable capacity of E. coli to adapt to environmental challenges, and have significant implications for the survival of spoilage and pathogenic bacteria, and hence for food safety.

Adaptation, Physiological↗

PubMed1997

Lipids and buoyancy in Southern ocean pteropods.

The lipids of Clione limacina, a Southern Ocean pteropod (order Gymnosomata), contain 28% diacylglyceryl ether (DAGE) (as percentage of total lipid) whereas the pteropod Limacina helicina (order Thecosomata) lacks DAGE. The alkyl glyceryl ether diols (1-O-alkyl glycerols, GE) of Clione DAGE are dominated by 16:0 (60%) and 15:0 (21%), in contrast with deep-sea shark liver DAGE, which is dominated by 18:1 GE. The fatty acid profiles of Clione and Limacina are similar (28-32% polyunsaturated, 26-34% monounsaturated) as are the sterols, which include 24-methylenecholesterol, transdehydrocholesterol, cholesterol, and desmosterol. This finding probably reflects the fact that Limacina is the major food source for Clione. Spongiobranchaea australis, another Southern Ocean pteropod (order Gymnosomata), has 0.9-1.7% DAGE, but has less lipid (3.3-4.8 mg/g lipid, wet weight) than Clione (50.8 mg/g lipid, wet weight). We propose a buoyancy role for DAGE in Clione since Limacina has bubbles for flotation which Clione lack; DAGE provides 23% more uplift than triacylglycerol at a concentration of 1.025 g/mL seawater.

Animals↗

PubMed1997

Psychroserpens burtonensis gen. nov., sp. nov., and Gelidibacter algens gen. nov., sp. nov., psychrophilic bacteria isolated from antarctic lacustrine and sea ice habitats.

Psychrophilic, yellow-pigmented, seawater-requiring bacteria isolated from the pycnocline of meromictic Burton Lake and from sea ice cores obtained in the Vestfold Hills (68 degrees S, 78 degrees E) in eastern Antarctica were characterized. Phenotypic analysis showed that the strains isolated formed two distinct taxa. The first taxon included nonmotile, nutritionally fastidious strains that were isolated from the pycnocline of Burton Lake. The cells of these strains were morphologically variant, ranging from vibrioid to ring shaped to coiled and filamentous; in addition, the strains were unable to metabolise carbohydrates or polysaccharides and had DNA G + C contents of 27 to 29 mol%. The strains of the second taxon, which were isolated from sea ice cores and from ice aigal biomass, were saccharolytic, exhibited rapid gliding motility, were rodlike to filamentous, and had DNA G + C contents of 36 to 38 mol%. A 16S ribosomal DNA (rDNA) sequence analysis revealed that the two Antarctic taxa formed related but distinct lineages within the [Flexibacter] maritimus rRNA branch of the family Flavobacteriacrae. The levels of 16S rDNA sequence similarity between the taxa were 90.5 to 91.3%, while the levels of similarity to other members of the [F.] maritimus rRNA branch were 85 to 90%. The whole-cell lipid profiles of the Antarctic strains were mainly comprised of branched and unbranched monounsaturated C15 to C17 fatty acids. The presence of significant levels of the lipids a 15:1 omega 10c and a17:1 omega 7c appeared to be useful biomarkers for the new Antarctic taxa and for differentiating these organisms from other members of the family Flavobacteriaceae. On the basis of polyphasic taxonomic data we propose that the new taxa are novel bacterial species designated Psychroserpens burtonensis gen. nov., sp. nov. (type strain, ACAM 188) and Gelidibacter algens gen. nov., sp. nov. (type strain, ACAM 536).

Antarctic Regions↗

PubMed1997

Shewanella gelidimarina sp. nov. and Shewanella frigidimarina sp. nov., novel Antarctic species with the ability to produce eicosapentaenoic acid (20:5 omega 3) and grow anaerobically by dissimilatory Fe(III) reduction.

A polyphasic taxonomic study was performed to characterize dissimilatory iron-reducing strains mostly isolated from Antarctic sea ice. The strains were isolated from samples of congelated (land-fast) sea ice, grease ice, and ice algal biomass collected from the coastal areas of the Vestfold Hills in eastern Antarctica (68 degrees S 78 degrees E). The strains were facultatively anaerobic, motile, and rod shaped, were capable of anaerobic growth either by fermentation of carbohydrates or by anaerobic respiration, and utilized a variety of electron acceptors, including nitrate, ferric compounds, and trimethylamine N-oxide. A phylogenetic analysis performed with 16S rRNA sequences showed that the isolates formed two groups representing novel lineages in the genus Shewanella. The first novel group included seawater-requiring, psychrophilic, chitinolytic strains which had DNA G + C contents of 48 mol%. The members of the second strain group were psychrotrophic and did not require seawater but could tolerate up to 9% NaCl. The strains of this group were also unable to degrade polysaccharides but could utilize a number of monosaccharides and disaccharides and had G + C contents of 40 to 43 mol%. The whole-cell-derived fatty acid profiles of the sea ice isolates were found to be similar to the profiles obtained for other Shewanella species. The omega-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) (20:5 omega 3) was detected in all of the sea ice isolates at levels ranging from 2 to 16% of the total fatty acids. EPA was also found at high levels in Shewanella hanedai (19 to 22%) and Shewanella benthica (16 to 18%) but was absent in Shewanella alga and Shewanella putrefaciens. On the basis of polyphasic taxonomic data, the Antarctic iron-reducing strains are placed in two new species, Shewanella frigidimarina sp. nov. (type strain, ACAM 591) and Shewanella gelidimarina sp. nov. (type strain, ACAM 456).

Antarctic Regions↗

PubMed1996

Microbial biomass in a shallow, urban aquifer contaminated with aromatic hydrocarbons: analysis by phospholipid fatty acid content and composition.

The city of Perth contains a number of sites that have been contaminated with hydrocarbons due to leakage from petroleum underground storage tanks. Microbial biomass in groundwater and sediment cores from above and below the water table, and from within and outside a plume of hydrocarbon contamination, was examined using phospholipid fatty acid methyl ester analysis. Microbial numbers, calculated from the phospholipid content, ranged from 0.9 x 10(6) to 7.8 x 10(6) 'Escherichia coli equivalent cells' g-1 dry wt of sediment. Over 96% of the microbial biomass was attached to the sediment and the proportion of attached cells did not decrease within the plume of contaminants. The amount of biomass within aquifer samples seemed to be related more to the proximity of the rhizosphere to the shallow aquifer, and other unknown urban inputs, rather than to the effects of the plume of contaminants. Fatty acids common to many bacterial groups dominated within the plume, and as such the analyses gave limited insight into microbial community structure. For site assessment of intrinsic remediation of shallow aquifers in urban areas, estimates of microbial biomass may not provide information that is readily applicable to plume management.

Biomass↗

PubMed1994

Phylogeny and lipid composition of Thermonema lapsum, a thermophilic gliding bacterium.

1,490 nucleotides of the 16S rRNA gene of a Gram-negative, thermophilic and gliding bacterium, Thermonema lapsum, have been sequenced. Phylogenetic analysis indicates that T. lapsum is related to cytophaga-flavobacteria-bacteroides (CFB) and is confirmed by the identification signature nucleotides that define this group. Further phylogenetic analysis indicates that T. lapsum forms the deepest branch in the CFB group; this observation was confirmed by the identification of unique nucleotide and nucleotide pairs which separate T. lapsum from all other members of this group. The phospholipid fatty acid (PLFA) profile also confirmed that T. lapsum is related to the cytophaga-flavobacteria-bacteroides group and also to selected members of the genus Flexibacter; the PLFA profile is unique to T. lapsum.

Bacteria, Aerobic↗

PubMed1993

Comparison of the abundance of the fecal sterol coprostanol and fecal bacterial groups in inner-shelf waters and sediments near Sydney, Australia.

Concurrent measurement of the sewage tracer coprostanol and fecal indicator bacteria were made for water and sediments collected in January 1992 from coastal waters off Sydney, Australia. The coprostanol results were compared with data from an earlier survey conducted in 1989 before the commisioning of Sydney's deepwater ocean outfalls in 1990 and 1991. Good correlations were observed for both water and sediment samples between coprostanol and the two fecal indicator organisms, fecal coliforms and Clostridium perfringens spores, thereby validating the use of coprostanol as a sewage signature in this environment. For sediments, most inner-shelf sites (1-10 km offshore) showed an increase in the concentration of coprostanol between the two surveys. The areas of highest concentration have been shifted further off-shore, to zones adjacent to the diffusers.

Australia↗

PubMed1986

Methyl sterol and cyclopropane fatty acid composition of Methylococcus capsulatus grown at low oxygen tensions.

Methylococcus capsulatus contained extensive intracytoplasmic membranes when grown in fed-batch cultures over a wide range of oxygen tensions (0.1 to 10.6%, vol/vol) and at a constant methane level. Although the biomass decreased as oxygen levels were lowered, consistently high amounts of phospholipid and methyl sterol were synthesized. The greatest amounts of sterol and phospholipid were found in cells grown between 0.5 and 1.1% oxygen (7.2 and 203 mumol/g [dry weight], respectively). While sterol was still synthesized in significant amounts in cells grown at 0.1% oxygen, the major sterol product was the dimethyl form. Analysis by capillary gas chromatography-mass spectrophotometry showed that the phospholipid esterified fatty acids were predominantly 16:0 and 16:1 and that the hexadecenoates consisted of cis delta 9, delta 10, and delta 11 isomers. At low oxygen tensions, the presence of large amounts (25%) of cyclopropane fatty acids (cy 17:0) with the methylene groups at the delta 9, delta 10, and delta 11 positions was detected. Although the delta 9 monoenoic isomer was predominant, growth at low oxygen levels enhanced the synthesis of the delta 10 isomers of 16:1 and cy 17:0. As the oxygen level was increased, the amount of cyclopropanes decreased, such that only a trace of cy 17:0 could be detected in cells grown at 10.6% oxygen. Although M. capsulatus grew at very low oxygen tensions, this growth was accompanied by changes in the membrane lipids.

Cyclopropanes↗

PubMed1986

A method for the separation and characterization of archaebacterial signature ether lipids.

A reproducible high performance liquid chromatography (HPLC) method for the separation of diethers and tetraethers isolated from archaebacterial phospholipids is reported. Fourier transform infrared spectroscopy was used for structural confirmation of these signature lipids. A mixture of tetraethers from a thermoacidophilic archaebacteria was resolved into three major components by the normal phase separation. These components were differentiated by Fourier self-deconvolution of infrared spectra. The application of the HPLC technique to environmental samples may provide an accurate assessment of archaebacterial biomass in various microbial communities.

Archaea↗

PubMed1985

Determination of monounsaturated double-bond position and geometry in the cellular fatty acids of the pathogenic bacterium Francisella tularensis.

The nonhydroxy fatty acid composition of Francisella tularensis is reported in detail. The double-bond configuration of the monounsaturated acids has been determined by capillary gas chromatography-mass spectrometry of the derivatized fatty acids. The monounsaturated fatty acids detected, in decreasing order of abundance, were 24:1 omega 15c, 18:1 omega 9c, 22:1 omega 13c, 20:1 omega 11c, 16:1 omega 7c, 26:1 omega 17c, and 14:1 omega 7c. The fatty acid profile found in F. tularensis, in particular the double-bond positions, represents a valuable taxonomic characteristic of this pathogenic bacterium.

Fatty Acids↗

PubMed1985

Extractable and lipopolysaccharide fatty acid and hydroxy acid profiles from Desulfovibrio species.

An analysis of the phospholipid ester-linked and the lipopolysaccharide (LPS) fatty acids and hydroxy fatty acids of six lactate-utilizing Desulfovibrio-type sulfate-reducing bacteria (SRB) has been performed using capillary gas-liquid chromatography-mass spectrometry (GLC-MS). The concentrations of normal fatty acids were essentially similar, with the possible exception of a high content of normal fatty acids in the LPS of Desulfovibrio gigas. Determination of monounsaturated acid double bond configuration was performed by GLC-MS analysis of the derivatized fatty acids. A total of nine branched chain and eight straight chain monounsaturated fatty acids was detected in the Desulfovibrio species analyzed. The major component detected in five Desulfovibrio was the 17-carbon iso-branched monoenoic acid which showed cis unsaturation [i17:1(n-7)c] seven carbons from the terminal methyl group of the fatty acid chain. D. gigas, in contrast, contained almost no unsaturated fatty acids and was greatly enriched in iso-branched 15:0. Major differences between strains were found in the phospholipid and LPS hydroxy fatty acids. These components, in addition to the i17:1(n-7)c and other characteristic branched chain unsaturated acids, can possibly be utilized as signatures of the lactate-utilizing SRB.

Desulfovibrio↗