Cellular fatty acid profiles for the differentiation of Penicillium species

Cellular fatty acid composition of eighteen species of Penicillium was studied to investigate its taxonomic usefulness. Many fungi included in this study displayed the same fatty acid composition but differed in relative concentration. Several test species presented the same fatty acid composition but differed in relative concentration. The principal fatty acids were palmitic (16:0), oleic (18:1) and linoleic (18:2). The amount of unsatured fatty acids varied between 68.5% and 78.5%. Multivariate analyses of data showed that it is possible to differentiate some species that belonged to different Penicillium groups. The level of agreement of long chain fatty acids with morphological taxonomy was acceptable.     

Chemotaxonomic characterization of rice seedling blight complex using fatty acid methyl ester (FAME) profiles

Rice seedling blight is an important disease caused by a complex of fungi that include Fusarium, Rhizopus, Pythium, and Trichoderma species. A modified MIDI method was used for extraction of fatty acids from these causal pathogens, and fatty acid methyl ester (FAME) profiles were characterized. Factors that might affect fatty acid production, such as period of culture and saponification in extraction, were also evaluated. A total of 14 fatty acids were detected, and FAME profiles showed quantitative and qualitative variations by discriminant analysis and principal component analysis. Genus-specific FAME profiles consisting of the types of fatty acid produced and remarkable components of individual fatty acids were observed. The possibility of application as chemotaxonomic methods based on the FAME profiles for diagnosis of the rice seedling blight complex is also discussed.     

The Fatty Acid Composition of Paramecium aurelia Cells and Cilia: Changes with Culture Age

SYNOPSIS.
The fatty acids of whole cells and cilia from Paramecium tetraurelia strains 51s and d,95 and from Paramecium octaurelia strain 299s were identified. Ciliates were grown axenically in 3 types of culture media. More than 30 fatty acid species were identified and their structures determined by gas chromatography, mass spectrometry, argentation chromatography, hydro-genation, and fragmentation technics. The major fatty acids were hexadecanoic, octadecanoic, 9-octadecenoic, 9,12-octadecadi-enoic, 6,9,12-octadecatrienoic, and 5,8,11,14-eicosatetraenoic acids. Minor variations in fatty acid compositions were observed in cells grown in the different culture media as well as among the 3 strains. Major changes in fatty acid compositions occurred with culture age and cell density. The cells accumulated exogenous lipids in cytoplasmic vesicles. These lipids were utilized as culture age progressed. Both cellular volume and lipid content were greater in young than in older cultures. Fatty acid compositions of both whole cells and cilia changed with age and had a relative decrease in saturated, short-chained and odd-numbered carbon acids. Cilia lipids were enriched in long-chained, polyunsaturated acids as compared to lipids in whole cell extracts. Eicosatetra-enoic acid (arachidonic acid) increased to the greatest extent with age in both cellular and ciliary lipids, accounting for 20–60% of the total fatty acids in cilia. The age-related change in fatty acid composition in Paramecium is among the largest observed in eukaryotic organisms. It was concluded that some minor fatty acids found in Paramecium lipids were incorporated directly from certain culture media and that Paramecium had w 3, 6, and 9 pathways for polyunsaturated fatty acid biosynthesis.     

Characterization of Aspergillus species based on fatty acid profiles

Cellular fatty acid (FA) composition was utilized as a taxonomic tool to discriminate between different Aspergillus species. Several of the tested species had the same FA composition and different relative FA concentrations. The most important FAs were palmitic acid (C16:0), estearic acid (C18:0), oleic acid (C18:1) and linoleic acid (C18:2), which represented 95% of Aspergillus FAs. Multivariate data analysis demonstrated that FA analysis is a useful tool for differentiating species belonging to genus Aspergillus. All the species analyzed showed significantly FA acid profiles (p < 0.001). Furthermore, it will be possible to distinguish among Aspergillus spp. in the Flavi Section. FA composition can serve as a useful tool for the identification of filamentous fungi.     

Marine fungi in sediments of the Sea of Japan (Russian territory) and their biologically active metabolites

The most abundant marine fungi encountered in various regions of the Sea of Japan belong to the genera Penicillium, Aspergillus, Wardomyces, Trichoderma, Chrysosporium, and Chaetomium. Facultative marine fungi of the genera Scytalidium, Verticillium, and Oidiodendron and obligate marine fungi of the genus Dendryphiella are much less abundant. The composition of marine sediments and the anthropogenic load on them were found to influence the abundance and species diversity of fungi, as well as the occurrence of fungal strains producing hemolytically active substances. The biodiversity of mycobiota and the abundance of hemotoxin-producing fungi in marine sediments may be used to evaluate the anthropogenic load on marine biocenoses. Hemolytic compounds were produced by 57% of the fungi isolated from marine sediments. The hemolytic activity of Chaetomium spiculipilium was revealed in the fraction of the culture liquid containing extracellular fatty acids and pigments. The fatty acid composition of this marine fungus was determined.     

Characterization of Flavobacterium species by analysis of volatile fatty acid production

Seventy-four Flavobacterium strains were characterized by gas-liquid chromatographic analysis of volatile fatty acids produced in the culture medium. Principal components analysis permitted the graphic representation of the relative positions of the different strains, and aggregation according to the variance enabled a hierarchical classification to be established. The study revealed three subgroups each for F. meningosepticum and F. odoratum. Our F. breve, Flavobacterium sp. group IIb and F. multivorum strains appeared to be homogeneous. These results tallied with those of previous studies on DNA base composition and reassociation, electrophoretic protein profiles and cellular fatty acid composition.     

Fatty acid metabolism of isolated mammalian cells

It is now clear that a wide variety of differentiated cells in culture exhibit essentially the full spectrum of mammalian fatty acid metabolism. These cells readily incorporate free fatty acids into membrane phosphoglycerides, modify exogenous fatty acids by desaturation and elongation, and store excess fatty acyl groups, primarily as triacylglycerols. Similarly, many different types of cells synthesize cyclooxygenase and lipoxygenase derivatives of long chain polyunsaturated fatty acids. Furthermore, although the fatty acid composition of cellular phospholipids can be modified by medium supplementation, cells in culture exhibit definite fatty acyl specificities for the various steps of fatty acid activation, transesterification and release. As the extensive repertoire of fatty acid metabolism in mammalian cells has been elucidated, and as the ability to grow differentiated cells in culture has increased, new questions have arisen. There is still much to be learned about the enzymes involved in synthesizing and maintaining the unique fatty acid composition of the different cellular phospholipids and the processes which regulate the desaturation, elongation and retroconversion of polyunsaturated fatty acids. Other areas of great current interest are the mechanisms by which certain long chain polyunsaturated fatty acids are made available for conversion to oxygenated, biologically-active derivatives, the metabolic interactions between different polyunsaturated fatty acids, particularly n-3 and n-6 fatty acids, the cellular roles of the C22 polyunsaturated fatty acids, and the functions of particular molecular species of phospholipids in membrane-mediated events. Further research in these areas will contribute to unravelling the role of fatty acids and fatty acid derivatives in the physiological processes of mammalian cells.     

Dependence of the fatty acid composition of the lipids in fungi of the genus Penicillium on their natural habitat conditions

The fatty acid composition of lipids was studied in Penicillium fungi growing in different habitats. Saturated fatty acids were represented by lauric, margaric, stearic and palmitic acids (the latter prevailed-- 18%-26%). Unsaturated monoene fatty acids were represented by acids from C16:1 to C18:1, diene and triene fatty acids by linoleic and linolenic acids. The predominance of linoleic acid was not found in all cultures of the genus. Changes in the fatty acid composition of lipids may be attributed to different ecological habitats of the Penicillium genus species.     

Polar lipid composition of leaves from nine typical alpine species

The fatty acids and polar lipid compositions of leaves from nine alpine species were almost identical to that of plants growing in habitats with little seasonal variation in temperature. Furthermore each polar lipid had about the same fatty acid composition in all plant species studied. It is suggested that neither the relative proportions of different lipid classes nor the degree of saturation of individual classes are directly implicated in the adaptation of plant tissues to different climates.     

INFLUENCE OF TEMPERATURE ON THE FATTY ACID PROFILE OF LISTERIA MONOCYTOGENES

Variation in the fatty acid profile of two Listeria monocytogenes strains grown at varying temperatures was determined. The fatty acid profiles varied greatly at different temperatures. General decreases in relative percentages of branched and medium chain (up to C16:0) fatty acids and variable changes in long chain fatty acids were found with increasing growth temperature. Individual fatty acid percentages between strains were variable. The relative percentages of unknown long chain fatty acids, detected in both strains at various temperatures, were greatest in Scott A (7.07%) and ATCC 19114 (13.15%) at 35C. Results demonstrated that L. monocytogenes had altered fatty acid profile in response to changes in growth temperature.     

Phospholipid,ester-linked fatty acid profiles as reproducible assays for changes in prokaryotic community structure of estuarine sediments

Phospholipid, ester-linked fatty acid profiles showed changes in benthic prokaryotic community structure reflecting culture manipulations that were both quantitative and statistically significant. Fatty acid structures, including the position and cis/ trans geometry of double bonds, were chemically verified by GC/MS after appropriate derivatization. The fatty acid profiles of independent flasks showed reproducible shifts when manipulated identically and significant differences when manipulated with different treatments. The absence of polyunsaturated fatty acids indicated that the consortia were predominantly prokaryotic. The prokaryotic consortia of different treatments could be differentiated by the proportions of cyclopropyl fatty acids and the proportions and geometry of monounsaturated fatty acids.     

Growth temperature affects accumulation of exogenous fatty acids and fatty acid composition in <Emphasis Type="Italic">Schizosaccharomyces pombe</Emphasis>

The incorporation of exogenously supplied fatty acids, palmitic acid, palmitoleic acid, oleic acid and linoleic acid, was examined in the yeast Schizosaccharomyces pombe at two growth temperatures, 20 °C and 30 °C. Fatty acids supplied to S. pombe in the growth medium were found to be preferentially incorporated into the cells, becoming a dominant species. The relative increase in exogenous fatty acids in cells came at the expense of endogenous oleic acid as a proportion of total fatty acids. Lowering the temperature at which the yeast were grown resulted in decreased levels of incorporation of the fatty acids palmitic acid, palmitoleic acid and linoleic acid compared to cells supplemented at 30 °C. In addition, the relative amount of the endogenously produced unsaturated fatty acid oleic acid, while greatly reduced compared to unsupplemented cells, was increased in cells supplemented with fatty acids at 20 °C compared to supplemented cells at 30 °C. The differential production of oleic acid in S. pombe cells indicates that regulation of unsaturated fatty acid levels, possibly by control of the stearoyl-CoA desaturase, is an important control point in membrane composition in response to temperature and diet in this species.     

Biochemical and Molecular Characterization of Some Centaurea Species Growing in the Eastern Anatolia Region of Turkey

Random amplified polymorphic DNA (RAPD) and fatty acid (FAME) profiles were used to examine phenotypic and genetic relationships among 16 Centaurea species growing wild in the eastern Anatolia region of Turkey. Thirteen decamer primers were used to examine polymorphism. According to the RAPD results, 99 amplicons in the size range of 50–1000 bp were produced from 13 primers in 16 Centaurea species. Genetically four distinct groups were determined among the species of Centaurea, which represents high genetic variation. In the 16 species, 14 fatty acids were determined according to FAME results. Both FAME and RAPD results showed that C. virgata is genetically different from other species. The differences in the composition of fatty acids among Centaurea species suggest that fatty acid profiles could be used to differentiate among some of these species. Results of this study show that RAPD and FAME analyses are consistent.     

ALTERATIONS IN LIPID MOLECULAR SPECIES OF THE MARINE EUSTIGMA TOPHYTE NANNOCHLOROSIS SP.1

The molecular species of triacylglycerol and monogalactosyl diacylglycerol from the marine eustigmatophyte Nannochloropsis were analyzed by high-performance liquid chromatography with a flame ionization detector. Four major molecular species of triacylglycerol composed of C 14:0, C 16:0, and C 16:1 fatty acids at different combinations were identified. Six molecular species of monogalactosyl diacylglycerol were detected. Three of them contained C 20:5 fatty acid in the sn-1 position, and one component accommodated C 20:5 fatty acid in both the sn-1 and sn-2 positions. Variations in the relative distribution of the molecular species were further monitored in Nannochloropsis cultures grown under different irradiance levels and temperatures. The relative distribution of 16: 0/16:1/16:0 triacylglycerol increased in cells grown in high light and in high temperature. Variations in cellular fatty acid composition in Nannochloropsis grown under different environmental conditions of irradiance level and temperature were attributed to alterations in relative cellular content of lipid classes as well as in the relative composition of lipid class molecular species.     

Analysis of interspecific variation in relative fatty acid composition: use of flow cytometry to estimate unsaturation index and relative polyunsaturated fatty acid content in microalgae

Relative polyunsaturated fatty acid content and unsaturation index are very important composition variables in the use of microalgae both for animal and human nutrition and biofuel production. A readily available technique to rapidly and inexpensively estimate relative fatty acid composition is very important for mass screening of new strains for the production of different types of oil. This study demonstrates the validity of Nile Red staining and flow cytometry for quick estimation of unsaturation index and relative fatty acid content in microalgae. Nile Red staining allows polar and neutral lipid contents to be estimated, and in this study a significant correlation was observed between polar/neutral ratio and fatty acid content in the species studied, corresponding to higher polyunsaturated fatty acid content in the polar lipid fraction of microalgae. This technique enables quick estimation of relative polyunsaturated fatty acid content and interspecific variation, as well as variations caused by culture conditions. In the species studied, most variability in fatty acid composition was due to variation in monounsaturated and polyunsaturated fatty acids, with less variation observed in saturated fatty acid content.     

Lipid Class and Fatty Acid Composition of the Protozoan Parasite of Oysters, Perkinsus marinus Cultivated in Two Different Media

The meront stage of the oyster protozoan parasite, Perkinsus marinus, cultivated in two media with different fatty acid profiles was analyzed for its fatty acid and lipid class composition. The composition of fatty acids in the prezoosporangium stage of the parasite as well as that of the host oyster were investigated. Although the lipid class composition of meronts was dominated by phospholipids and triacylglycerol, there was no triaclgycerol detected in either culture medium. Despite the difference in fatty acid composition of the two media, the fatty acid composition of meronts in each medium was dominated by 14:0, 16:0, 18:0, 18:1(n-9), 20: (n-9), 18:2(n-6) and 20:4(n-6), a profile that differed from its host. The quantities of total lipids and fatty acids in meronts increased as the number of meronts increased and far exceeded the initial amounts in the media and in the initial cell inoculum. The meronts harvested 25 d post-inoculation, had about 3 to 6 times higher total lipids and 4 to 13 times higher fatty acids than the amounts contained in the media. The fatty acid profiles of both prezoosporangia and oysters resembled each other and consisted primarily of 16:0, 20:4(n-6), 20:5(n-3), 22:2delta7,15, and 22:6(n-3). These results indicate that during meront proliferation, the parasite synthesizes certain fatty acids and lipid classes. For development from meront to prezoosporangium, the parasite may rely on its host for lipid resources.     

Phospholipids of the differentiating species of Tetrahymena

The whole-cell phospholipid composition of the six known polymorphic species of Tetrahymena has been examined by [(3)H]acetate and [(3)H]myristic acid radiolabeling, and by gas-liquid chromatography of total phospholipid-bound fatty acids. Five of the polymorphic species contained similar phospholipid profiles following radiolabeling in that phosphatidylethanolamine (PE) was the predominant phospholipid; however, in cells of Tetrahymena patula LFF, aminoethylphosphonolipid was present in amounts nearly equal to PE. Tetrahymena patula LFF contained an unusually large percentage of sphingolipid (16.2% by [(3)H]acetate radiolabeling). Substantial differences were found in the fatty acid profiles of the polymorphic species, which included the degree of fatty acid unsaturation and relative weight percentages of odd-chain fatty acids. Tetrahymena vorax contained a low ratio of unsaturated C(18) fatty acids to saturated C(18) fatty acids as compared with all other species examined. The differentiating species generally contained a lesser percentage of monoenoic fatty acids and a lower ratio of unsaturated C(16) fatty acids to saturated C(16) fatty acids as compared with the two monomorphic species examined.     

Discrimination of Sebastes viviparus, Sebastes marinus and Sebastes mentella from Faroe Islands by chemometry of the fatty acid profile in heart and gill tissues and in the skull oil

The composition of fatty acids in the tissue of heart, gill and skull oil of the three redfish species, Sebastes viviparus, Sebastes marinus and Sebastes mentella was determined by a chemometric method. The method consists of methanolysis of samples of the tissues and of the oils, gas chromatography of the resulting fatty acid methyl esters and multivariate statistical treatment, by principal component analysis, of the analytical data. Although the differences in fatty acid composition among the three tissues were the dominating features of the data, the three species had significantly different fatty acid profiles within each tissue, even though variation among the individuals was considerable. The fatty acid profiles appear to be species specific. The mutual relationship between S. marinus and S. mentella is closer than the relationship between either of them and S. viviparus.     

Fungi in sediments of the sea of Japan and their biologically active metabolites

The most abundant marine fungi encountered in various regions of the Sea of Japan belong to the generaPenicillium, Aspergillus, Wardomyces, Trichoderma, Chrysosporium, andChaetomium. Facultative marine fungi of the generaScytalidium, Verticillium, andOidiodendron and obligate marine fungi of the genusDendryphiella are much less abundant. The composition of marine sediments and the anthropogenic load on them were found to influence the abundance and species diversity of fungi, as well as the occurrence of fungal strains producing hemolytically active substances. The biodiversity of mycobiota and the abundance of hemotoxin-producing fungi in marine sediments may be used to evaluate the anthropogenic load on marine biocenoses. Hemolytic compounds were produced by 57% of the fungi isolated from marine sediments. The hemolytic activity ofChaetomium spiculipilium was revealed in the fraction of the culture liquid containing extracellular fatty acids and pigments. The fatty acid composition of this marine fungus was determined.     

Cuticular fatty acid profile analysis of three Rhipicephalus tick species (Acari: Ixodidae)

Cuticular fatty acids (CFA) are important constituents of the arthropod exoskeleton, serving as structural and defense components, and participating in intra-species communication. Here we describe for the first time a comparative analysis of the CFA profiles of three tick species of the genus Rhipicephalus: R. annulatus, R. bursa and R. sanguineus. CFA profiles were determined for R. bursa and R. sanguineus grown both on rabbit or calf, and for R. annulatus grown on calf. CFA composition was compared for each species before and after ethanol treatment, for different hosts of each species, and between the different species. Our data suggest that adsorption of the host’s fatty acids changes the apparent CFA composition. Ethanol treatment efficiently removed the unbound fatty acids from the ticks and revealed the actual composition. Comparison between ticks grown on rabbit versus calf showed significant difference in the relative abundance of fatty acids C14 and 9,12-C18:2 for R. bursa, and a difference in the relative abundance of C14 for R. sanguineus. Comparison of the CFA between the three species revealed significant differences in the abundance of fatty acids C16, 9,12-C18:2, 9-C18:1, C18 and C20. Our results show that while the host had a minor effect on CFA composition within each species, significant differences were observed in the CFA profiles of different species. We suggest that CFA profiles may be used to distinguish between related species. CFA analysis can also be used in studies of communication and defense mechanisms in ticks and other arthropods.