Variability in fatty acids and fatty aldehydes in different organs of two prosobranch gastropod mollusks

The distribution of fatty acids and fatty aldehydes of total lipids in mantle, foot and digestive gland of two prosobranch gastropod mollusks, Bellamya bengalensis and Pila globosa, from India were studied. The individual volatile compounds, e.g., acids and aldehydes, were separated by serially coupled capillary columns with different polarity stationary phase and identified by gas chromatography-mass spectrometry. Saturated fatty acids were the main components and vary from 48–60%, monoenic are 18–30%, while polyunsaturates vary between 21–33%. The digestive glands of both the snail species contained cyclopropane fatty acids, not previously reported in gastropods.     

Fatty Acid Profiling of Commercially Important Raw and Boiled Seer Fish <Emphasis Type="Italic">Scomberomorus commerson</Emphasis> (Lacepede, 1800) (Scombridae,Teleostei, Pisces)

The present study aims to determine the fatty acid profiling of commercially important fresh and boiled Scomberomorus commerson. Fatty acids in fresh and boiled fish were separated and quantitatively determined by gas chromatography–mass spectrophotometer using standard methods. The findings revealed that the predominant fatty acids in fresh S. commerson were octadecanoic acid methyl ester, octanoic acid, 1,2-benzenedicarboxylic acid and 3-cyclopentylpropionic acid representing respectively 45.91, 5.69, 6.75 and 8.65% of total fatty acids. Boiled S. commerson showed predominant changes in their fatty acid profiles. In the omega-3 and omega-6 families the dominant fatty acids were doconexent, 3-cyclopentylpropionic acid, octadeconoic acid methyl ester and Hexadecane representing respectively 3.87, 12.08, 44.26 and 3.11% of total fatty acids. After boiling, some fatty acids present in fresh fish are damaged and formed new fatty acids which belonged to ω-3 polyunsaturated fatty acids (PUFA). Boiling increased the concentration of PUFAs from 73.25 to 80.37% of total fatty acids and also formed new fatty acids.     

Injection principles in capillary gas chromatographic analysis of bacterial fatty acids

Principles for sample injection in fused silica capillary gas chromatographic analysis of bacterial cellular fatty acids are discussed. Comparative analyses were made of the methyl esters of fatty acids in a reference solution and in Legionella pneumophila serogroup 1, using both split and splitless injection. A splitless injection technique, in which the methyl esters are cold-trapped immediately after injection and consecutively released in accordance with temperature programming, gave sensitivity increase corresponding to the split ratio used in split injections, without diminishing the speed of analysis or separation efficiency. Splitless injection, utilizing cold-trapping, can be recommended for capillary gas chromatography in routine profiling of bacterial fatty acids.     

Fatty acid composition in the classification of Saccharopolyspora hirsuta

The long-chain fatty acids of 4 Saccharopolyspora hirsuta strains were examined as their methyl and picolinyl esters using gas liquid chromatography (GLC) and GLC-mass spectrometry (GLC-MS). All the strains had similar fatty acid profiles composed mainly of isoanteiso-, and 10-methyl-branched components. Three new substituted 10-methyl-branched fatty acids were also detected and the major component identified as 10,15-dimethylhexadecanoic acid. The implications of these data for Saccharopolyspora systematics are discussed.     

Aliphatic Hydrocarbons and Fatty Acids of Some Marine and Freshwater Microorganisms

Gas chromatography and combined gas chromatography-mass spectrometry have been used to study the fatty acids and hydrocarbons of a bacterium from the Pacific Ocean, Vibrio marinus, a freshwater blue-green alga, Anacystis nidulans, and algal mat communities from the Gulf of Mexico. Both types of microorganisms (bacteria and algae) showed relatively simple hydrocarbon and fatty acid patterns, the hydrocarbons predominating in the region of C-17 and the fatty acids in the range of C-14 to C-18. The patterns of V. marinus were more comparable to those of the algal populations than to patterns reported for other bacteria. An incomplete correlation between fatty acids and hydrocarbons in both types of organisms was observed, making it difficult to accept the concept that the biosynthesis of hydrocarbons follows a simple fatty acid decarboxylation process.     

A two-step extraction procedure for concentrating acidic organic volatiles in aqueos solution prior to gas chromatographic head-space analysis,as exemplified by short-chain fatty acids produced by Bacillus cereus

A concentration procedure for short-chain fatty acids in aqueos solution is described, utilizing extraction by diethyl ether followed by re-extraction of fatty acid salts from the ether phase into a small amount of aqueous NaOH. The samples were acidified and analyzed by automatic head-space gas chromatography using a fused silica capillary column. The method's usefulness for trace analysis of short-chain fatty acids was exemplified in a study on broth cultures of Bacillus cereus where acetic, isobutyric and isovaleric acids could be readily detected after only 12 h of incubation.     

Lipid Metabolism of Rumen Ciliates and Bacteria: II. Uptake of Fatty Acids and Lipid Analysis of Isotrichia intestinalis and Rumen Bacteria with Further Information on Entodinium simplex

The total lipid and free fatty acid contents of Isotricha intestinalis, Entodinium simplex, and the rumen bacterial flora of the respective protozoa were determined. Warburg manometric data showed that the sodium salts of tributyrin, oleic, and acetic acids stimulated gas production in I. intestinalis, whereas tributyrin was stimulatory with E. simplex and less active with oleic and acetic acids. Rumen bacteria provided fatty acids produced lower manometric gaseous increases when compared with the protozoa. Volatile fatty acids were produced by I. intestinalis and rumen bacteria with tributyrin, but not with tripalmitin. Sodium oleate gave little volatile fatty acid response with I. intestinalis or rumen bacteria. Washed suspensions of I. intestinalis and rumen bacteria concentrated C¹⁴-labeled oleic, palmitic, stearic, and linoleic acids within the cells during short incubation periods. Autoradiographs demonstrated the conversion of C¹⁴-labeled oleic, palmitic, stearic, linoleic, and acetic acids in the rumen protozoa and bacterial cells.     

Dietary Supplementation of Polyunsaturated Fatty Acids in Caenorhabditis elegans

Fatty acids are essential for numerous cellular functions. They serve as efficient energy storage molecules, make up the hydrophobic core of membranes, and participate in various signaling pathways. Caenorhabditis elegans synthesizes all of the enzymes necessary to produce a range of omega-6 and omega-3 fatty acids. This, combined with the simple anatomy and range of available genetic tools, make it an attractive model to study fatty acid function. In order to investigate the genetic pathways that mediate the physiological effects of dietary fatty acids, we have developed a method to supplement the C. elegans diet with unsaturated fatty acids. Supplementation is an effective means to alter the fatty acid composition of worms and can also be used to rescue defects in fatty acid-deficient mutants. Our method uses nematode growth medium agar (NGM) supplemented with fatty acidsodium salts. The fatty acids in the supplemented plates become incorporated into the membranes of the bacterial food source, which is then taken up by the C. elegans that feed on the supplemented bacteria. We also describe a gas chromatography protocol to monitor the changes in fatty acid composition that occur in supplemented worms. This is an efficient way to supplement the diets of both large and small populations of C. elegans, allowing for a range of applications for this method.     

Fatty acids ofLactobacillus bulgaricus determined by gas chromatography — Mass spectrometry

Fatty acids (in the form of pyrrolidinides) ofLactobacillus bulgaricus were identified by gas chromatography—mass spectrometry using two columns differing in the polarity of the stationary phase. In addition to the described fatty acids (primarily the most abundant vaccenic acid), we detected nine fatty acids that have not yet been described in the genusLactobacillus (all of them minority compounds) and in four of them we determined the position of the double bond. The occurrence of vaccenic acid was related to other sources of positional isomers of octadecenoic acids (parsley seed - petroselmic acid; olive oil — oleic acid).     

Quality evaluation of Semen persicae (seed of Prunus persica L. Batsch) by gas chromatography–mass spectrometry

Seeds of Prunus persica L. Batsch were collected at various times from different regions of China. The fatty acid composition of the seeds was analysed using gas chromatography-mass spectrometry (GC-MS). The aim was to evaluate whether genetic and environmental factors influenced the chemical profile of fatty acids in Semen persicae. Saturated and unsaturated fatty acids were present at 4.8-8.7% and 90.7-94.8% of total fatty acids, respectively. Oleic and linoleic acids were dominant in all samples and ranged from 59.3 to 81.4% and from 11.6 to 31.0%, respectively. All samples had high levels of unsaturated fatty acids. Hierarchical clustering analysis and principal components analysis were performed to differentiate and classify the samples based on the contents of the characteristic fatty acid constituents. This study provides evidence that metabolites may reflect genetic and environmental similarities, such as management practices used in cultivars (irrigation, fertilization and sanitary treatments), and evidence of the variability of genetic make up and local environmental conditions in samples grown in the wild.     

Evaluation of fatty acid composition among selected amaranth grains grown in two consecutive years

The oil of amaranth grain (Amaranthus spp.) is a rich source of poly-unsaturated fatty acids. In this study, we tested 10 amaranth samples representing two species (Amaranthus cruentus and Amaranthus hypochondriacus) in two consecutive years (2010, 2011). Grain oils were analysed by gas chromatography for their fatty acids profile. In 2010, oil content ranged from 6.4–8.2% for A. cruentus and 6.3–7.9% for A. hypochondriacus. In 2011, the level was 7.1–8.2% and 6.6–8.7% for A. cruentus and A. hypochondriacus, respectively. Linoleic, palmitic, and oleic acids were dominant fatty acids in all of the oil samples. The essential linoleic acid level was 33.3–38.7% (A. cruentus) and 31.7–47.5% (A. hypochondriacus) in 2010 and 34.6–39.9% (A. cruentus) and 34–44.5% (A. hypochondriacus) in 2011. The minority fatty acids, i.e. stearic, α-linolenic, and arachidic acids were also observed. Eicosenoic and behenic acids were present in the grain in trace amounts. Statistical evaluation showed a significant effect of year and species of amaranth on the levels of certain fatty acids. There was a strong positive correlation between oil content and oleic acid, and a negative correlation between oleic acid and either of the other two fatty acids, linoleic and α-linolenic ones.     

Survey of Extreme Solvent Tolerance in Gram-Positive Cocci: Membrane Fatty Acid Changes in Staphylococcus haemolyticus Grown in Toluene

We exploited the unique ecological niche of oil fly larval guts to isolate a strain of Staphylococcus haemolyticus which may be the most solvent-tolerant gram-positive bacterium yet described. This organism is able to tolerate 100% toluene, benzene, and p-xylene on plate overlays and saturating levels of these solvents in monophasic liquid cultures. A comparison of membrane fatty acids by gas chromatography after growth in liquid media with and without toluene showed that in cells continuously exposed to solvent the proportion of anteiso fatty acids increased from 25.8 to 33.7% while the proportion of 20:0 straight-chain fatty acids decreased from 19.3 to 10.1%. No changes in the membrane phospholipid composition were noted. Thus, S. haemolyticus alters its membrane fluidity via fatty acid composition to become more fluid when it is exposed to solvent. This response is opposite that commonly found in gram-negative bacteria, which change their fatty acids so that the cytoplasmic membrane is less fluid. Extreme solvent tolerance in S. haemolyticus is not accompanied by abnormal resistance to anionic or cationic detergents. Finally, six strains of Staphylococcus aureus and five strains of Staphylococcus epidermidis, which were not obtained by solvent selection, also exhibited exceptional solvent tolerance.     

The attractiveness of odorous esterified fatty acids to the potential biocontrol agent,Altica cyanea

The fatty acid composition of foliar buds, young, mature, and senescent leaves, and stem parts of the rice-field weed, Ludwigia adscendens L. (Onagraceae) was analyzed by thin layer chromatography and gas chromatography flame ionization detection. The analysis of fatty acid composition revealed that saturated fatty acids (i.e., C14:0, C16:0, and C18:0) were prevailing compounds among the all weed parts except senescent leaves where C18:1 was predominant. The esterified fatty acids isolated from different weed parts over the range of 10–100 μg/ml followed by individual synthetic esterified fatty acids that were identified from the esterified extracts of different weed parts, and a mixture of synthetic esterified fatty acids except esterified eicosenoic acid and docosahexaenoic acid were applied to identify their role as a chemical cue for a potential biocontrol agent, Altica cyanea (Weber) (Coleoptera: Chrysomelidae) in a Y-tube olfactometer under laboratory conditions. In this bioassay, the esterified fatty acids from mature leaves and stem parts of this weed attracted A. cyanea at 20–100 μg/ml and at 80 μg/ml concentrations, respectively. Clear attraction was recorded by female A. cyanea insects in the mixture of synthetic esterified fatty acids at 60, 80, and 100 μg/ml concentrations. It is thus concluded that A. cyanea rely on an effective proportion of esterified fatty acids as an olfactory cue for attraction.     

Anaerobic n-Alkane Metabolism by a Sulfate-Reducing Bacterium, Desulfatibacillum aliphaticivorans Strain CV2803T

The alkane-degrading, sulfate-reducing bacterium Desulfatibacillum aliphaticivorans strain CV2803^T, recently isolated from marine sediments, was investigated for n-alkane metabolism. The total cellular fatty acids of this strain had predominantly odd numbers of carbon atoms (C odd) when the strain was grown on a C-odd alkane (pentadecane) and even numbers of carbon atoms (C even) when it was grown on a C-even alkane (hexadecane). Detailed analyses of those fatty acids by gas chromatography/mass spectrometry allowed us to identify saturated 2-, 4-, 6-, and 8-methyl- and monounsaturated 6-methyl-branched fatty acids, with chain lengths that specifically correlated with those of the alkane. Growth of D. aliphaticivorans on perdeuterated hexadecane demonstrated that those methyl-branched fatty acids were directly derived from the substrate. In addition, cultures on pentadecane and hexadecane produced (1-methyltetradecyl)succinate and (1-methylpentadecyl)succinate, respectively. These results indicate that D. aliphaticivorans strain CV2803^T oxidizes n-alkanes into fatty acids anaerobically, via the addition of fumarate at C-2. Based on our observations and on literature data, a pathway for anaerobic n-alkane metabolism by D. aliphaticivorans is proposed. This involves the transformation of the initial alkylsuccinate into a 4-methyl-branched fatty acid which, in addition to catabolic reactions, can alternatively undergo chain elongation and desaturation to form storage fatty acids.     

Olfactory responses of Epilachna dodecastigma (Coleoptera: Coccinellidae) to long-chain fatty acids from Momordica charantia leaves

Extraction, thin-layer chromatography and gas chromatography–mass spectrophotometry analyses revealed the presence of 12, 13, and 12 fatty acids in young, mature, and senescent leaves of Momordica charantia L., representing 87.30, 95.25, and 83.11 % of the total fatty acids, respectively. The proportion of saturated fatty acids was highest in senescent leaves (78.60 %) followed by young leaves (69.42 %) and mature leaves (48.92 %), with the balance accounted for by unsaturated fatty acids. Palmitic acid was the predominant saturated fatty acid in the three types of leaves, whereas alpha-linolenic acid was the predominant unsaturated fatty acid. The fatty acids from young, mature, and senescent leaves followed by the application of a synthetic mixture of fatty acids that was comparable to the natural fatty acids found in the three types of leaves, elicited the attraction of the female insect Epilachna dodecastigma (Coleoptera: Coccinellidae) at 50–200, 50–200, and 100–200 μg/ml concentrations, respectively, in a Y-shaped glass tube olfactometer bioassay. Individual synthetic fatty acids were also evaluated by the olfactometer bioassay at concentrations comparable to the proportions detected in the three types of leaves. Individual synthetic palmitic acid, stearic acid, oleic acid, linoleic acid, and alpha-linolenic acid at 58.24, 13.96, 29.40, 30.31, and 29.76 μg, respectively, attracted the insect. A synthetic blend of 79.13, 10.57, 29.40, 30.31, and 36.33 μg of palmitic, stearic, oleic, linoleic, and alpha-linolenic acids, respectively, which is the proportion present in a 200 μg/ml concentration of fatty acids of mature leaves, or of 116.49, 13.96, and 29.76 μg of palmitic, stearic and alpha-linolenic acids, respectively, which is the proportion present in a 200 μg/ml concentration of natural fatty acids of young leaves, served as attractants for E. dodecastigma.     

Stable-Isotope-Based Labeling of Styrene-Degrading Microorganisms in Biofilters

Deuterated styrene ([²H₈]styrene) was used as a tracer in combination with phospholipid fatty acid (PLFA) analysis for characterization of styrene-degrading microbial populations of biofilters used for treatment of waste gases. Deuterated fatty acids were detected and quantified by gas chromatography-mass spectrometry. The method was evaluated with pure cultures of styrene-degrading bacteria and defined mixed cultures of styrene degraders and non-styrene-degrading organisms. Incubation of styrene degraders for 3 days with [²H₈]styrene led to fatty acids consisting of up to 90% deuterated molecules. Mixed-culture experiments showed that specific labeling of styrene-degrading strains and only weak labeling of fatty acids of non-styrene-degrading organisms occurred after incubation with [²H₈]styrene for up to 7 days. Analysis of actively degrading filter material from an experimental biofilter and a full-scale biofilter by this method showed that there were differences in the patterns of labeled fatty acids. For the experimental biofilter the fatty acids with largest amounts of labeled molecules were palmitic acid (16:0), 9,10-methylenehexadecanoic acid (17:0 cyclo9-10), and vaccenic acid (18:1 cis11). These lipid markers indicated that styrene was degraded by organisms with a Pseudomonas-like fatty acid profile. In contrast, the most intensively labeled fatty acids of the full-scale biofilter sample were palmitic acid and cis-11-hexadecenoic acid (16:1 cis11), indicating that an unknown styrene-degrading taxon was present. Iso-, anteiso-, and 10-methyl-branched fatty acids showed no or weak labeling. Therefore, we found no indication that styrene was degraded by organisms with methyl-branched fatty fatty acids, such as Xanthomonas, Bacillus, Streptomyces, or Gordonia spp.     

Rapid microdetermination of fatty acids in biological materials by gas-liquid chromatography

Total and free fatty acids in general ranging from lauric to nervonic acid were separated and quantitated based on an internal standard method as methyl esters by “on column” methylation with trimethyl-(α,α,α-trifluoro-m-tolyl) ammonium hydroxide (TMTFTH) in a gas chromatographic system. This study represents an application of a method published by MacGee and Allen and a change to an internal standard technique. For the determination of the total fatty acids the sampls were saponified with KOH-CH₃OH, acidified with H₂PO₄, and then the fatty acids were extracted into hexane. An aliquot of the hexane extract was then extracted with TMTFTH and chromatographed. For determination of free fatty acids the sample was acidified with H₃PO₄, immediately extracted with hexane and processed as described earlier. The relative standard deviation of 1.4 to 4.2% illustrates the precision of the method and the recovery of the fatty acids ranged from 88.5 to 100.5%. This method was applied to the determination of fecal fatty acids in conjunction with an interdepartmental study on “High protein diet in colon cancer” at the University of Missouri. In addition, the applicability of the analytical procedure (with small modifications) was shown for a wide variety of biological materials (serum, milk, skin tissue, fungal spores, food homogenates, beef tissues, and tumor cell cultures). The analyses were performed on different gas chromatographs by different analysts.     

In Vitro Study and Comparison of Caecal Methanogenesis and Fermentation Pattern in the Brown Hare (Lepus europaeus) and Domestic Rabbit (Oryctolagus cuniculus)

The brown hare and the domestic rabbit are mid-sized herbivorous mammals and hindgut fermenters, though their digestive physiologies differ in some traits. The objective of this study was to estimate and compare the caecal microbial activity in hares and rabbits via an analysis of the following end-products of in vitro caecal fermentation: methane, total gas production, short chain fatty acids and ammonia concentration. Hare caecal methanogenesis occurred at a much lower level (0.25 mmol/kg for samples incubated without substrate and 0.22 mmol/kg for samples incubated with substrate) than that of the rabbit (15.49 and 11.73 mmol/kg, respectively) (P<0.001). The impact of the substrate’s presence on caecal methanogenesis was not significant, though its presence increased the total gas production during fermentation (P<0.001). Hare caecal microflora produced a lower short chain fatty acids concentration than did rabbit microorganisms (P<0.05). In unincubated hare samples, the short chain fatty acids concentration was 28.4 mmol/kg, whereas in unincubated rabbit samples, the short chain fatty acids concentration was 51.8 mmol/kg. The caecal fermentation pattern of the hare was characterised by higher propionate and isobutyrate molar proportions compared with those observed in rabbit caecum (P<0.01). No significant changes in the ammonia concentration in either rabbit or hare caecum were found. The results obtained indicate some differences in the activity of the microbial populations colonising the hare and rabbit caecum, particularly in regards to methanogenic Archaea.     

Long-chain free fatty acids from Momordica cochinchinensis leaves as attractants to its insect pest,Aulacophora foveicollis Lucas (Coleoptera: Chrysomelidae)

Extraction, thin layer chromatography, and gas chromatography–mass spectrometry of young, mature, and senescent leaves of Momordica cochinchinensis Spreng revealed 13 free fatty acids, representing a total of 82.29, 91.30, and 68.52% of fatty acids in young, mature, and senescent leaves, respectively. Palmitic acid was the predominant fatty acid followed by stearic acid in three types of leaves. The free fatty acids from young, mature, and senescent leaves attracted female Aulacophora foveicollis Lucas (Coleoptera: Chrysomelidae) at the minimal concentrations of 4, 2, and 8 μg, respectively; whereas the mixtures of synthetic fatty acids mimicking free fatty acids of young, mature, and senescent leaves showed attraction at the minimal concentrations of 4, 2, and 10 μg, respectively, in Y-shaped glass tube olfactometer bioassay under laboratory condition. The results indicate that A. foveicollis may employ long-chain free fatty acids as an olfactory cue for host location. The individual synthetic fatty acids mimicking the proportions detected in three types of leaves were also evaluated through olfactometer bioassay. Only synthetic palmitic acid at the minimal amount of 2.17 μg attracted the insect. A synthetic blend of fatty acids mimicking 8 μg free fatty acid concentration of mature leaves or an amount of 5.42 μg palmitic acid produced the highest attraction of the insect. Hence 5.42 μg palmitic acid might be used for insect pest management program such as baited traps.     

Incorporation of bacterial fatty acids and changes in a wax ester-based omnivory index during a long-term incubation experiment with Calanus glacialis Jaschnov

We traced the incorporation of fatty acid biomarkers into Calanus glacialis Jaschnov (CV) during a long-term incubation experiment using bacterivorous dinoflagellates and diatoms as food. Copepods fed Oxyrrhis marina Dujardin during a 3-week acclimation period developed an omnivorous lipid composition, relative to wild-captured copepods, characterized by significant losses of polyunsaturated fatty acids (PUFA) and diatom fatty acids [16:4(n−1), 20:5(n−3)], and increases in saturated fatty acids (SFA) and 18:1(n−7). Levels of a wax ester-based omnivory index [unsaturation coefficient (UC)], verified by gas chromatography (GC), also decreased in response to the relatively PUFA-poor dinoflagellate. After half of the copepods were switched to a diet comprised of the diatom Thalassiosira hispida Syvertsen (PUFA-rich), the data showed reversal to a more herbivorous lipid composition (increases in UC and relative amounts of PUFA and diatom fatty acids). We assert that UC, derived from routine thin-layer chromatography analysis (Iatroscan) can quickly determine in situ feeding strategies (i.e., degree of omnivory) of wax ester-storing copepods. None of the eight odd and/or branched fatty acids (OBFA) initially detected in C. glacialis increased in response to a diet of O. marina which was rich in these compounds [mainly iso (i)-15:0 and anteiso (ai)-15:0]. Lack of transfer of these and other fatty acids [e.g., 22:6(n−3)] could be related to the physiological state of the copepods (early diapause). We suggest that the bacterial fatty acid 18:1(n−7) may be more useful in inferring connections between Calanus spp. and the microbial food web than the odd and/or branched chains.