Substrate specificity in hydrocarbon utilizing microorganisms

Three bacteria (designated stram JOB5, 7E4, and 7E1C) isolated from soil by elective culture techniques and capable of growth on a wide variety of hydrocarbons were tested for substrate specificity. Non-proliferating cells of strain JOB5 grown on each of the C₁ to C_N series of normal ahphatic hydrocarbons were assayed for the capacity to oxidize all the alkanes, alcohols, fatty acids, and methyl-ketones in the homologous series of straight chain compounds. Cells of strain 7E4 grown on the gaseous alkanes (C₁ C₄) and strain 7E1C grown on propane were tested for the ability to oxidize the C₁ to C₄ n-alkanes, alcohols, and fatty acids.Contribution from Microbiology, North Carolina Agricultural Experiment Station, Raleigh, North Carolina Published with the approval of the Director of Research as paper No 2395 of the Journal Series.Part of this work was done while the author was associated with the late Dr. Jackson W. Foster, at the University of Texas, Austin.     

A renewable energy source — hydrocarbon gases resulting from pyrolysis of the marine nanoplanktonic alga Emiliania huxleyi

The marine coccolithophore, Emiliania huxleyi, grown in the laboratory was subjected to vacuum pyrolysis at various temperatures from 100 to 500 °C. The highest yield of pyrolytic gases (183 mL g⁻¹ dry cells) was obtained at 400 °C. The amount of total hydrocarbon gas produced at 400 °C was 129 mL, about 10 times higher than at 300 °C. CH₄ was the major component at the high gas-production stage (400–500 °C). The great increase in hydrocarbon gases at 400 °C was accompanied by a marked decrease in liquid saturates and aromatics. The results indicate that the liquid hydrocarbons (oil) produced by pyrolysis at lower temperature is a direct source for the formation of the hydrocarbon gases. Due to its large potential for the production of biomass and hydrocarbons with low energy input, E. huxleyi is suggested as one of candidates for the production of renewable fuels. This revised version was published online in July 2006 with corrections to the Cover Date.     

2010秋季东海今生颗石藻的空间分布

根据2010年11月东海29站位所采集108个样品偏光镜检分析结果显示:调查区共发现26种今生颗石藻,优势种为赫氏艾密里藻(Emiliania huxleyi)、大洋桥石藻(Gephyrocapsa oceanica)、卡特螺旋球藻(Helicosphaera carteri)和粗壮环翼球(Algirosphaera robusta)等.水体中今生颗石藻丰度为0-76.562个/mL,平均值为18.641个/mL;颗石粒丰度0-4506.47个/mL,平均值为613.44个/mL,今生颗石藻在表层和底层均呈斑块状分布,海盆区站位丰度较近岸站位下降明显,但物种丰富度明显增多;颗石球丰度最大值站位发生在硅藻水华站位.将研究区域划分为3个断面进行分析:PN断面、沿岸流断面和黑潮断面.比较显示,PN断面的空间分布差异性较大,赫氏艾密里藻具有最高丰度的颗石粒和颗石球.沿岸流断面颗石粒/颗石球、黑潮断面中颗石粒(除底层外),自表至底随水深增加有轻微增加趋势,黑潮断面颗石球丰度高值区集中于25-80 m之间.结合历史环境资料,分析得出东海调查期水体混合、泥沙再悬浮以及硝酸盐浓度是控制今生颗石藻物种丰富度与物种特异分布的主要因子;今生颗石藻在不同环境中具有不同的生活策略,其空间分布特征随取样季节和海区不同有较大变化.     

INTRASPECIFIC GENETIC DIVERSITY IN THE MARINE COCCOLITHOPHORE EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE): THE USE OF MICROSATELLITE ANALYSIS IN MARINE PHYTOPLANKTON POPULATION STUDIES1

Using primer pairs for seven previously described microsatellite loci and three newly characterized microsatellite loci from the coccolithophore Emiliania huxleyi (Lohm.) Hay and Mohler, we assessed genetic variation within this species. Analysis of microsatellite length variants (alleles) was conducted for 85 E. huxleyi isolates representative of different ocean basins. These results revealed high intraspecific genetic variability within the E. huxleyi species concept. Pairwise comparison of a 1992 Coastal Fjord group (FJ92) (n=41) and a North East Atlantic (NEA) group (n=21), using F_ST as an indicator of genetic differentiation, revealed moderate genetic differentiation (F_ST=0.09894; P=0; significance level=0.05). Gene flow between the FJ92 and NEA groups was estimated to be low, which is in agreement with the moderate levels of genetic differentiation revealed by the microsatellite data. A genetic assignment method that uses genotype likelihoods to draw inference about the groups to which individuals belong was tested. Using FJ92 and NEA as reference groups, we observed that all the E. huxleyi groups tested against the two reference groups were unrelated to them. On a global biogeographical scale, E. huxleyi populations appear to be highly genetically diverse. Our findings raise the question of whether such a high degree of intraspecific genetic diversity in coccolithophores translates into variability in ecological function.     

LIPID BIOMARKER DIVERSITY IN THE COCCOLITHOPHORID EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE) AND THE RELATED SPECIES GEPHYROCAPSA OCEANICA1

Twenty-four strains of Emiliania huxleyi and two strains of Gephyrocapsa oceanica were grown at 15°C under identical culture conditions to assess genetic variability in key lipid biomarker profiles (C₃₇-C₃₉ alkenones, C₃₆ and C₃₇ alkyl alkenoates, and C₃₁-C₃₈ alkenes). Under our culture conditions, little divergence an biomarker composition was detected between E. huxleyi strains from different oceanic regions or between E. huxleyi and G. oceanica even though the strains originated from biogeographical regions as diverse as the subpolar North Atlantic and subtropical Western Pacific. The major differences observed were in tetraunsaturated alkenone abundance and alkene profiles, which tended to separate neritic from open ocean strains. Different strains from the same locality were as different as strains originating from widely separated ocean basins, indicating extreme genotypic diversity within a population. Replicate cultures of the same strain showed significant variability in their biomarker profiles even though the culture temperature varied by only ±0.3°C, indicating that their synthesis ratios are influenced by environmental and/or physiological variable(s), as yet unidentified, in addition to temperature. Strong covariance in C₃₇ and C₃₈ methyl alkenone unsaturation ratios (U^k₃₇ and U^k_38Mρ respectively) and, in coastal strains, C₃₃, alkene and alkenone unsaturation ratios indicates that these compounds are biochemically linked.     

Biodegradation of Petroleum Hydrocarbons in Seawater at Low Temperatures (0–5 °C) and Bacterial Communities Associated with Degradation

In this study biodegradation of hydrocarbons in thin oil films was investigated in seawater at low temperatures, 0 and 5 °C. Heterotrophic (HM) or oil-degrading (ODM) microorganisms enriched at the two temperatures showed 16S rRNA sequence similarities to several bacteria of Arctic or Antarctic origin. Biodegradation experiments were conducted with a crude mineral oil immobilized as thin films on hydrophobic Fluortex adsorbents in nutrient-enriched or sterile seawater. Chemical and respirometric analysis of hydrocarbon depletion showed that naphthalene and other small aromatic hydrocarbons (HCs) were primarily biodegraded after dissolution to the water phase, while biodegradation of larger polyaromatic hydrocarbons (PAH) and C₁₀–C₃₆ n-alkanes, including n-hexadecane, was associated primarily with the oil films. Biodegradation of PAH and n-alkanes was significant at both 0 and 5°C, but was decreased for several compounds at the lower temperature. n-Hexadecane biodegradation at the two temperatures was comparable at the end of the experiments, but was delayed at 0°C. Investigations of bacterial communities in seawater and on adsorbents by PCR amplification of 16S rRNA gene fragments and DGGE analysis indicated that predominant bacteria in the seawater gradually adhered to the oil-coated adsorbents during biodegradation at both temperatures. Sequence analysis of most DGGE bands aligned to members of the phyla Proteobacteria (Gammaproteobacteria) or Bacteroidetes. Most sequences from experiments at 0°C revealed affiliations to members of Arctic or Antarctic consortia, while no such homology was detected for sequences from degradation experiment run at 5°C. In conclusion, marine microbial communities from cold seawater have potentials for oil film HC degradation at temperatures ≤5°C, and psychrotrophic or psychrophilic bacteria may play an important role during oil HC biodegradation in seawater close to freezing point.     

Protein expression in the obligate hydrocarbon-degrading psychrophile Oleispira antarctica RB-8 during alkane degradation and cold tolerance

In cold marine environments, the obligate hydrocarbon-degrading psychrophile Oleispira antarctica RB-8, which utilizes aliphatic alkanes almost exclusively as substrates, dominates microbial communities following oil spills. In this study, LC–MS/MS shotgun proteomics was used to identify changes in the proteome induced during growth on n-alkanes and in cold temperatures. Specifically, proteins with significantly higher relative abundance during growth on tetradecane (n-C₁₄) at 16°C and 4°C have been quantified. During growth on n-C₁₄, O. antarctica expressed a complete pathway for the terminal oxidation of n-alkanes including two alkane monooxygenases, two alcohol dehydrogenases, two aldehyde dehydrogenases, a fatty-acid-CoA ligase, a fatty acid desaturase and associated oxidoreductases. Increased biosynthesis of these proteins ranged from 3- to 21-fold compared with growth on a non-hydrocarbon control. This study also highlights mechanisms O. antarctica may utilize to provide it with ecological competitiveness at low temperatures. This was evidenced by an increase in spectral counts for proteins involved in flagella structure/output to overcome higher viscosity, flagella rotation to accumulate cells and proline metabolism to counteract oxidative stress, during growth at 4°C compared with 16°C. Such species-specific understanding of the physiology during hydrocarbon degradation can be important for parameterizing models that predict the fate of marine oil spills.     

TESTING THE EFFECTS OF ELEVATED PCO2 ON COCCOLITHOPHORES (PRYMNESIOPHYCEAE): COMPARISON BETWEEN HAPLOID AND DIPLOID LIFE STAGES1

The response of Emiliania huxleyi (Lohmann) W. W. Hay et H. Mohler, Calcidiscus leptoporus (G. Murray et V. H. Blackman) J. Schiller, and Syracosphaera pulchra Lohmann to elevated partial pressure of carbon dioxide (pCO₂) was investigated in batch cultures. We reported on the response of both haploid and diploid life stages of these three species. Growth rate, cell size, particulate inorganic carbon (PIC), and particulate organic carbon (POC) of both life stages were measured at two different pCO₂ (400 and 760 parts per million [ppm]), and their organic and inorganic carbon production were calculated. The two life stages within the same species generally exhibited a similar response to elevated pCO₂, the response of the haploid stage being often more pronounced than that of the diploid stage. The growth rate was consistently higher at elevated pCO₂, but the response of other processes varied among species. Calcification rate of C. leptoporus and of S. pulchra did not change at elevated pCO₂, whereas it increased in E. huxleyi. POC production and cell size of both life stages of S. pulchra and of the haploid stage of E. huxleyi markedly decreased at elevated pCO₂. It remained unaltered in the diploid stage of E. huxleyi and C. leptoporus and increased in the haploid stage of the latter. The PIC:POC ratio increased in E. huxleyi and was constant in C. leptoporus and S. pulchra. Elevated pCO₂ has a significant effect on these three coccolithophore species, the haploid stage being more sensitive. This effect must be taken into account when predicting the fate of coccolithophores in the future ocean.     

Components of natural gas resulting from thermal degradation of the blue-green alga (cyanobacterium)Oscillatoria tenuis

Cultures of the blue-green alga (cyanobacterium)Oscillatoria tenuis were used to simulate thermal degradation and gas formation by heating without oxygen at 250° and 350 °C for 100 h. Analysis through gas chromatography showed that the gases were mainly CH₄, C₂H₆, C₃H₈, iC₄ (isobutane), nC₄ (normal butane), iC₅ (isopentane), nC₅ (normal pentane), H₂, C0₂ and N₂. The volume of gases per g dry weight of alga was 44 ml at 250 °C and 100 ml at 350 °C. Alkane gas comprised only 2.04% of the total at 250 °C and rising to 40.0% at 350 °C. The fraction of C0₂ decreased from 83.3% at 250 °C to 40.0% at 350 °C. The quantity of alkane in the soluble organic matter doubled with rising temperature but the H/C atomic ratio in the ‘kerogen’, insoluble organic matter, decreased sharply. Infrared spectra of the ‘kerogen’ showed that the peak of adipose radical at 2900 cm⁻¹ disappeared gradually with rising temperature, which reflects the gradual break of CH₄ or C₂H₆ from ‘kerogen’. This demonstrates that insoluble organic matter rather than soluble organic matter in blue-green algae are the main sources of the gas alkanes in the process of simulated thermal degradation.     

OCCURRENCE OF ALKANES IN BRAIN MYELIN. COMPARISON BETWEEN NORMAL AND QUAKING MOUSE

Alkanes, a new class of neurolipid, were found in mouse brain, the level being reduced in the Quaking mutant. These hydrocarbons are concentrated in myelin; minor amounts being found in microsomes, mitochondria and synaptosomes. The average recovery is 7.1 μg/mg in normal myelin, 2.2 in the Quaking myelin. The distribution pattern of these alkanes was determined by gas liquid chromatography and was found to differ in normal and Quaking myelin; the hydrocarbons consist mainly of n-alkanes ranging from C₂₁ to C₃₂ with even and odd aliphatic chains.     

New insight into the contributions of thermogenic processes and biogenic sources to the generation of organic compounds in hydrothermal fluids

Experiments on hydrothermal degradation of Pyrococcus abyssi biomass were conducted at elevated pressure (40 MPa) over a 200–450 °C temperature range in sapphire reaction cells. Few organic compounds could be detected in the 200 °C experiment. This lack was attributed to an incomplete degradation of P. abyssi cells. On the contrary, a wide range of soluble organic molecules were generated at temperatures ≥350 °C including toluene, styrene, C₈–C₁₆ alkyl‐benzenes, naphthalene, C₁₁–C₁₆ alkyl‐naphthalenes, even carbon number C₁₂–C₁₈ polycyclic aromatic hydrocarbons, C₁₅–C₁₈ alkyl‐phenanthrenes and C_8:0–C_16:0 n‐carboxylic acids. The effect of time on the final organic composition of the degraded P. abyssi solutions at 350 °C was also investigated. For that purpose the biomass was exposed for 10, 20, 60, 90, 270 and 720 min at 350 °C. We observed a similar effect of temperature and time on the chemical diversity obtained. In addition, temperature and time increased the degree of alkylation of alkyl‐benzenes. This study offers additional evidence that a portion of the aliphatic hydrocarbons present in the fluids from the Rainbow ultramafic‐hosted hydrothermal field may be abiogenic whereas a portion of the aromatic hydrocarbons and n‐carboxylic acids may have a biogenic origin. We suggest that aromatic hydrocarbons and linear fatty acids at the Rainbow site may be derived directly from thermogenic alteration of material from the sub‐seafloor biosphere. Yet we infer that the formation and dissolution of carboxylic acids in hydrothermal fluids may be controlled by other processes than in our experiments.     

Macromolecular composition of Palaeozoic scolecodonts: insights into the molecular taphonomy of zoomorphs

Dutta, S., Hartkopf‐Fröder, C., Mann, U., Wilkes, H., Brocke, R. & Bertram, N. 2010: Macromolecular composition of Palaeozoic scolecodonts: insights into the molecular taphonomy of zoomorphs. Lethaia, Vol. 43, pp. 334–343. Biogeochemistry and molecular taphonomy of biopolymers of marine zoomorphs are poorly known. In order to obtain insights into this issue we report on the biogeomacromolecular composition of hand‐picked, well‐preserved scolecodonts of Ordovician, Silurian and Devonian age using micro‐Fourier transform infrared (micro‐FTIR) spectroscopy, Curie point pyrolysis‐gas chromatography‐mass spectrometry (Cupy‐GC‐MS) and tetramethylammonium hydroxide (TMAH)‐assisted thermochemolysis‐GC‐MS. The present study reveals that scolecodonts are composed of both aliphatic and aromatic moieties. The micro‐FTIR spectra of scolecodonts are characterized by aliphatic CH_x (3000–2800 and 1460–1450/cm) and CH₃ (1375/cm) absorptions and aromatic C=C (1560–1610/cm) and CH (3050/cm and 700–900/cm) absorptions. The major pyrolysis products from the scolecodonts include aromatic hydrocarbons such as alkylbenzenes, alkylnaphthalenes and alkylphenols. Aliphatic hydrocarbons are represented by a homologous series of n‐alkenes and n‐alkanes. The compounds released upon thermochemolysis with TMAH are saturated and unsaturated fatty acids (as their methyl esters), n‐alkenes/alkanes and aromatic acids (as their methyl esters). No protein/amino acid‐derived compounds have been recognized in the pyrolysates or in the thermochemolysates, and it is concluded that protein/amino acid‐related compounds, which are commonly found in the jaws of extant polychaetes, were destroyed due to diagenetic processes. Obviously, excellent morphological preservation and low thermal alteration are not paralleled by a similar degree of chemical preservation. □Biogeomacromolecules, micro‐FTIR, pyrolysis‐GC‐MS, scolecodonts, thermochemolysis‐GC‐MS.     

Sex- and age-related changes in the biophysical properties of cuticular lipids of the housefly,Musca domestica

We examined the biophysical properties of cuticular lipids isolated from the housefly, Musca domestica. Melting temperatures (T_m) of surface lipids isolated from female houseflies decreased from 39.3 °C to 35.3 °C as the females attained sexual maturity and produced sex pheromone, whereas those prepared from males did not change with age. Lipids melted over a 10–25 °C temperature range, and their physical properties were a complex function of the properties of the component lipids. The T_m of total cuticular lipids was slightly below that of cuticular hydrocarbons (HC), the predominant lipid fraction. Hydrocarbons were further fractionated into saturated, unsaturated, and methyl-branched components. The order of decreasing T_m was total alkanes > total HCs > methyl-branched alkanes > alkenes. For 1-day-old flies, measured T_ms of hydrocarbons were 1.3–5.5 °C lower than T_ms calculated from a weighted average of T_ms for saturated and unsaturated components. For 4-day-old flies, calculated T_ms underestimated T_m by 11–14 °C. © 1995 Wiley-Liss, Inc.     

Production of alkanes (C7–C29) from different part of poplar tree via direct deoxy-liquefaction

Poplar leaves, poplar bark and poplar wood were deoxy-liquefied directly in an air-proof stainless steel reactor at different temperatures. The oils from leaves at 350 °C, from bark at 400 °C and from wood at 450 °C, at which the liquid product yields were the maximum, were analyzed by GC–MS. The oils obtained from three parts of poplar tree were quite different from each other in the relative contents of their compositions. The oil from leaves was rich in hydrocarbons (alkanes: C₇–C₂₉; aromatics) and poor in phenolics, while oil from wood was rich in phenolics and poor in hydrocarbons. The oil from bark was moderate. Relative contents of hydrocarbons in the leaves oil were as high as 60.01% but decreased to 29.71% in bark oil and 11.43% in wood oil. GC analysis of gases and FT-IR, GC–MS and elemental analysis of oils were performed in this study.     

Aerobic Biodegradation of Crude Oil Components by Acidophilic Mycobacteria

Biodegradation of crude oil components by strain AG_S10, an acidophilic member of the genus Mycobacterium, was studied under extremely acidic conditions (pH 2.5). The degree of degradation of the same hydrocarbons in different kinds of oil was found to be different. The direction of biodegradation was, however, the same: the share of n-alkanes in oxidized oil decreased, while the share of branched alkanes increased. At the same time, the degree of redistribution of methane hydrocarbons in degraded oil varied significantly for different oils, although no strict dependence on the type of oil was found. After 28 days of incubation at 30°C and pH 2.5, the degradation of n- and iso-alkanes was 99 and 44%, respectively for the light, low-viscosity oil of the Nizhnevartovsk deposit, 58 and 32%, respectively for the medium-density oil of the Moscow oil-procesing plant, and 80 and 16% and 99 and 69%, respectively for the heavy, viscous oils of the Cheremukhovskoe and Usinkoye oil fields. Moreover, after extended cultivation time strain AG_S10 completely utilized alkanes, as well as a significant part of the naphthene component of the aliphatic fraction. The studied strain was characterized by ability to oxidize a broad spectrum of methane hydrocarbons, including high-molecular C₁₇–C₃₀n-alkanes, in oils of different properties and composition. Apart from its scientific interest, farther investigation of biodegradation of high-paraffin oils and viscous oils with elevated paraffin content by strain AG_S10 may be useful in view of the technical issues associated with paraffin accumulation in the course of recovery and transportation of these oils.     

Biomarker indications for microbial contribution to Recent and Late Jurassic carbonate deposits

Summary Biomarker investigations were applied to the hydrocarbon fractions of three Recent (cyanobacterial mat, Lake Van microbialite and Lake Satonda microbialite) and two Late Jurassic carbonate samples obtained from sponge bioherms. The relative concentrations ofn-alkanes, monomethyl alkanes, acyclic isoprenoids, steroids and hopanoids in these samples are studied and their probable biological precursors are discussed. Normal alkanes with carbon chain lengths ranging from C₁₅ to C₃₄ and monomethyl alkanes ranging from C₁₇ to C₂₁ with a varying methyl branching pattern are found. The major hydrocarbons are low molecular weight (LMW)n-alkanes (C₁₅–C₂₁) with a slight to strong predominance ofn-heptadecane (C₁₇). High molecular weight (HMW)n-alkanes occur in low to moderate relative concentrations showing a preference of odd-carbon numbered compounds with a maximum at C₂₉. Within the acyclic isoprenoids, pristane, phytane/phytene, pentamethyl-eicosane, squalane and lycopane could be identified. Polycyclic terpenoids of the sterane and/or hopane type are present in all carbonate samples. The carbon numbers of these components range from 27 to 29 and 27 to 32, respectively. These organic compounds identified can be attributed to various source organisms such as cyanobacteria, archaebacteria, algae and vascular plants. All hydrocarbon fractions of the samples are characterized by moderate to high relative concentrations of compounds derived from cyanobacteria, signifying the role of these organisms as contributors to the Recent as well as to the Late Jurassic carbonate deposits.     

Stable isotope fractionation of strontium in coccolithophore calcite: Influence of temperature and carbonate chemistry

Marine calcifying eukaryotic phytoplankton (coccolithophores) is a major contributor to the pelagic production of CaCO₃ and plays an important role in the biogeochemical cycles of C, Ca and other divalent cations present in the crystal structure of calcite. The geochemical signature of coccolithophore calcite is used as palaeoproxy to reconstruct past environmental conditions and to understand the underlying physiological mechanisms (vital effects) and precipitation kinetics. Here, we present the stable Sr isotope fractionation between seawater and calcite (Δ^88/86Sr) of laboratory cultured coccolithophores in individual dependence of temperature and seawater carbonate chemistry. Coccolithophores were cultured within a temperature and a pCO₂ range from 10 to 25°C and from 175 to 1,240 μatm, respectively. Both environmental drivers induced a significant linear increase in coccolith stable Sr isotope fractionation. The temperature correlation at constant pCO₂ for Emiliania huxleyi and Coccolithus braarudii is expressed as Δ^88/86Sr = ?7.611 × 10^?3 T + 0.0061. The relation of Δ^88/86Sr to pCO₂ was tested in Emiliania huxleyi at 10 and 20°C and resulted in Δ^88/86Sr = ?5.394 × 10^?5 pCO₂ – 0.0920 and Δ^88/86Sr = ?5.742 × 10^?5 pCO₂ – 0.1351, respectively. No consistent relationship was found between coccolith Δ^88/86Sr and cellular physiology impeding a direct application of fossil coccolith Δ^88/86Sr as coccolithophore productivity proxy. An overall significant correlation was detected between the elemental distribution coefficient (D_Sr) and Δ^88/86Sr similar to inorganic calcite with a physiologically induced offset. Our observations indicate (i) that temperature and pCO₂ induce specific effects on coccolith Δ^88/86Sr values and (ii) that strontium elemental ratios and stable isotope fractionation are mainly controlled by precipitation kinetics when embedded into the crystal lattice and subject to vital effects during the transmembrane transport from seawater to the site of calcification. These results provide an important step to develop a coccolith Δ^88/86Sr palaeoproxy complementing the existing toolbox of palaeoceanography.     

Studies on the Low Temperature Fermentation

A marine yeast, strain MM313 was isolated from a marine sediment sample at depth of 1120 m. The organism was identified as a Candida sp. MM313. The yeast was able to utilize n-paraffin, n-C₁₀ to n-C₂₀. Regardless of its origin, the organism grew in a medium prepared with fresh water. However, the cell yield increased with increasing concentration of each salt in sea water in the medium and reached a maximum value at the concentration of 75%. The cultivation temperature for the maximum rate of growth and that for the maximum level of growth were 28° and 10°C, respectively. Several cultural conditions were investigated. The cell yields to n-paraffins were about 85% at 15°C after 4 days and 56% at 28°C after 3 days under optimal conditions.     

Degradation of long-chain n-alkanes (C36 and C40) by Pseudomonas aeruginosa strain WatG

Pseudomonas aeruginosa strain WatG was unable to utilize either n-hexatriacontane (C₃₆) or n-tetracontane (C₄₀), which are both insoluble in a mineral salts medium (MSM), as a sole carbon source. However, when C₃₆ and C₄₀ were added to MSM containing crude oil, more than 25% of each of the compounds was degraded by this strain after 2 weeks at 30 °C. These results demonstrate that P. aeruginosa strain WatG has the ability to degrade long-chain alkanes up to C₄₀, when they are solubilized by crude oil components.     

CELL AND GROWTH CHARACTERISTICS OF TYPES A AND B OF EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE) AS DETERMINED BY FLOW CYTOMETRY AND CHEMICAL ANALYSES1

Two morphotypes of Emiliania huxleyi (Lohmann 1902) Hay et al. 1967, types A and B, known to be unequally distributed in the oceans, were grown in dilution cultures at a range of photon flux densities (PFDs) (1.5–155 μmol photons·m^?2·s^?1) and two temperatures (10° and 15° C). Calcite carbon and organic carbon content of the cells as well as instantaneous growth rate, cell size, chlorophyll fluorescence, and light-scatter properties clearly depended on growth conditions and differed considerably for the two morphotypes. The ratio between calcite carbon and organic carbon production showed an optimum of 0.65 in E. huxleyi type A cells at PFD = 17.5. The ratio increased slightly with a temperature increase from 10° to 15°C but remained < 1.0 at both temperatures in light-limited cells. In contrast, calcite carbon production exceeded organic carbon production (ratio: 1.4–2.2) in phosphate-deprived cultures. Emiliania huxleyi type B generally showed a higher calcite carbon/organic carbon ratio than E. huxleyi type A, but the relation with PFD was similar. The content of calcite carbon and organic carbon as well as the instantaneous growth rate, cell size, chlorophyll fluorescence, and light-scatter properties showed large diel variations that were closely related to the division cycle. Our results show the importance of mapping the structure of any sampled cell population with respect to the phase in the cell division cycle, as this largely determines the outcome of not only “per cell” measurements but also short time (less than 24 h) flux measurements. For instance, dark production of calcite by E. huxleyi was negatively affected by cell division. Slowly growing (phosphate-stressed) cultures produced calcite in the light and in the dark. In contrast, rapidly growing cultures at 10°C produced calcite only in the light, whereas in the dark there was a significant loss of calcite due to dissolution.