Typologie de bactéries prélevées devant le delta du Rhǒne. Utilisation d'une méthode objective: L'analyse des correspondances

Bacteria isolated from surface sea waters, brackish waters and from algal cultures were classified in nutritional clusters. Results obtained with an hierarchical method were improved by using the correspondence analysis. The former main statements appear confirmed and new informations are pointed out, mainly concerning the organic compounds which are the best indexes of the nutritional characteristics; e. g. a few organic acids and amino-acids can be used to estimate the versatility of bacteria; it also appears that strains which preferentially assimilate substrates of one chemical group, do not easily attack substances of other groups.     

Bactéries hétérotrophes aérobies prélevées devant le delta du rhône. II — Détermination des exigences nutritionnelles. Comparaison avec des souches provenant d'autres biotopes

Résumé Les auteurs comparent un lot de 114 bactéries prélevées dans des eaux eutrophes situées devant le delta du Rhône à deux lots de souches appartenant également au bactérioplancton méditerranéen, le premier prélevé dans des eaux oligotrophes (200 souches), le deuxième constitué par des germes épiphytes-saprophytes d'algues planctoniques (57 souches).Plus de 50% des bactéries épiphytes-saprophytes ont des besoins vitaminiques; peu d'entre elles se développent sur le glucose et les acides organiques; la plupart exigent des milieux complexes. Au contraire, les souches de pleine eau présentent un grand pouvoir de synthèse qui leur permet de croître sur des substrats simples.Dans l'ensemble, le glucose n'est pas un substrat carboné universellement assimilé en présence d'azote minéral, au contraire des acides aminés, qui sont rapidement et efficacement utilisés. Cette utilisation préférentielle des acides aminés serait plus marquée pour les bactéries des eaux oligotrophes que pour celles des eaux eutrophes.

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Aerobic heterotrophic bacteria isolated in front of the Rhône outlet. II — Nutritional requirements. Comparison with strains from different origin.One hundred and fourteen bacteria have been isolated from eutrophic surface sea waters sampled in front of the Rhône outlet (Mediterranean Sea). Their vitamins requirements as a whole and growth potentials were studied with the West and Lochead's method. Results are compared with those obtained with 200 strains isolated from oligotrophic surface waters and 57 strains living epiphytically or saprophytically on diatoms sampled offshore.It appears that more than 50% of the epiphytic-saprophytic bacteria require vitamins; a few grow on glucose and organic acids, most need complex media. On the opposite, simple organic substances support the growth of free-living bacterioplankton; this capability obviously results from their high synthetizing power.Data support the opinion that the glucose should not be considered as an usual source of carbon for the bacteria studied as a whole, when a mineral nitrogen source is present. On the contrary, the amino-acids sustain fast growth. Nevertheless this preferential use is more related to the strains sampled in the oligotrophic waters than to those of eutrophic waters.

     

Stable carbon isotope analysis of nucleic acids to trace sources of dissolved substrates used by estuarine bacteria.

The natural abundance of stable carbon isotopes measured in bacterial nucleic acids extracted from estuarine bacterial concentrates was used to trace sources of organic matter for bacteria in aquatic environments. The stable carbon isotope ratios of Pseudomonas aeruginosa and nucleic acids extracted from cultures resembled those of the carbon source on which bacteria were grown. The carbon isotope discrimination between the substrate and total cell carbon from bacterial cultures averaged 2.3% +/- 0.6% (n = 13). Furthermore, the isotope discrimination between the substrate and nucleic acids extracted from bacterial cultures was 2.4% +/- 0.4% (n = 10), not significantly different from the discrimination between bacteria and the substrate. Estuarine water samples were prefiltered through 1-micron-pore-size cartridge filters. Bacterium-sized particles in the filtrates were concentrated with tangential-flow filtration and centrifugation, and nucleic acids were then extracted from these concentrates. Hybridization with 16S rRNA probes showed that approximately 90% of the nucleic acids extracted on two sample dates were of eubacterial origin. Bacteria and nucleic acids from incubation experiments using estuarine water samples enriched with dissolved organic matter from Spartina alterniflora and Cyclotella caspia had stable carbon isotope values similar to those of the substrate sources. In a survey that compared diverse estuarine environments, stable carbon isotopes of bacteria grown in incubation experiments ranged from -31.9 to -20.5%. The range in isotope values of nucleic acids extracted from indigenous bacteria from the same waters was similar, -27.9 to -20.2%. Generally, the lack of isotope discrimination between bacteria and nucleic acids that was noted in the laboratory was observed in the field.(ABSTRACT TRUNCATED AT 250 WORDS)     

Degradation of urea by bacteria and algae in mass algal cultures.

Studies were conducted to determine the role of bacteria and algae in the degradation of urea in industrial waste waters. The microflora accompanying algae in continuous cultures on waste waters from the nitrogen fertilizers industry was found to include bacteria capable of intensive degradation of urea. Urea hydrolyzing bacteria are with time eliminated from the algal culture. This elimination can be attributed to algal metabolites which inhibit the development of sensitive bacteria. The Chlorella vulgaris strain used in the treatment of wast waters from the nitrogen fertilizers industry did not hydrolyse urea.     

THE ANTIBACTERIAL SUBSTANCES OF THE MARINE ALGA STICHOCHRYSIS IMMOBILIS (CHRYSOPHYTA)12

Two substances with antibacterial activity have been concentrated from the culture medium of Stichochrysis immobilis before senescence occurred, and purified, by a Sephadex G-25 filtration followed by 2-dimensional thin-layer chromatography on Merck F-254 silica gel plates. One of the substances is obviously a peptide; 11 different amino acids are linked in a 30 amino acid molecule (MW 3400). It was not possible to assign a chemical group to the second substance (MW ? 2600); it may be related to “humic acids” (Kalle's gelbstoff). Growth of the most sensitive bacteria was completely inhibited by 20-fold, or higher, concentrations of substances from the algal culture filtrate (assuming no loss of substances during the concentration processes). The 54 strains of test bacteria were of various origins, though mostly isolated from algal cultures or obtained directly from the marine environment.     

Stable carbon isotope analysis of nucleic acids to trace sources of dissolved substrates used by estuarine bacteria

The natural abundance of stable carbon isotopes measured in bacterial nucleic acids extracted from estuarine bacterial concentrates was used to trace sources of organic matter for bacteria in aquatic environments. The stable carbon isotope ratios of Pseudomonas aeruginosa and nucleic acids extracted from cultures resembled those of the carbon source on which bacteria were grown. The carbon isotope discrimination between the substrate and total cell carbon from bacterial cultures averaged 2.3% +/- 0.6% (n = 13). Furthermore, the isotope discrimination between the substrate and nucleic acids extracted from bacterial cultures was 2.4% +/- 0.4% (n = 10), not significantly different from the discrimination between bacteria and the substrate. Estuarine water samples were prefiltered through 1-micron-pore-size cartridge filters. Bacterium-sized particles in the filtrates were concentrated with tangential-flow filtration and centrifugation, and nucleic acids were then extracted from these concentrates. Hybridization with 16S rRNA probes showed that approximately 90% of the nucleic acids extracted on two sample dates were of eubacterial origin. Bacteria and nucleic acids from incubation experiments using estuarine water samples enriched with dissolved organic matter from Spartina alterniflora and Cyclotella caspia had stable carbon isotope values similar to those of the substrate sources. In a survey that compared diverse estuarine environments, stable carbon isotopes of bacteria grown in incubation experiments ranged from -31.9 to -20.5%. The range in isotope values of nucleic acids extracted from indigenous bacteria from the same waters was similar, -27.9 to -20.2%. Generally, the lack of isotope discrimination between bacteria and nucleic acids that was noted in the laboratory was observed in the field.(ABSTRACT TRUNCATED AT 250 WORDS)     

Bactéries hétérotrophes aerobies prélevées devant le delta du rhone. III. utilisation potentielle de differents substrats organiques comme source de carbone

Résumé Dans le cadre d'une étude portant sur un lot de 200 bactéries prélevées en surface dans des eaux eutrophes au débouché du Rhône en Méditerranée, les auteurs de carbone de 100 composés organiques appartenant à différentes classes chimiques.Dans l'ensemble, il est apparu que les familles de substances les mieux assimilées sont, par ordre décroissant, les acides gras, utilisés par 55% des souches, les acides aminés aliphatiques (51%), les acides aminés divers (44%), les hydroxyacides (43%), les acides dicarboxyliques (42%), les acides organiques divers (30%), les sucres (28%), les dérivés des sucres (27%), les alcools (21%) et les bases (19%).La plupart des souches étudiées utilisent un nombre élevé de substrats, mais aucun substrat n'apparaît comme universel.A partir de ces résultats et de ceux fournis par la littérature, les auteurs font remarquer que la proline, le glutamate, le pyruvate et le succinate sont les substrats les plus fréquemment utilisés, à la fois par les bactéries du plancton et par celles des sédiments côtiers et profonds. Ils en concluent que ces composés pourraient remplacer avantageusement le glucose lors des mesures du taux d'assimilation hétérotrophe des bactéries in situ.

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Aerobic heterotrophic bacteria isolated in front of the Rhône outlet. III. Organic substrates supporting growth.Two hundred strains of bacteria have been isolated from eutrophic surface waters; the substrates used as the sole source of carbon have been screened within 100 organic compounds belonging to different chemical groups.The fatty acids are the most frequently used substances, being attacked by 55% of the strains. The other compounds are ordered as follows: aliphatic amino-acids (51%), miscellaneous amino-acids (44%), hydroxyacids (43%), dicarboxylic acids (42%), miscellaneous organic acids (30%), carbohydrates (28%), carbohydrates derivatives (27%), alcohols (2i%) and organic bases (19%).Most of the strains attack numerous substrates, but none of these substrates is used by all the bacteria.From the data reported and from those in the literature, it can be stated that prolin, glutamate, pyruvate and succinate are attacked by most of the bacteria sampled both from surface waters and from neritic or deep sea sediments. It is pointed out that the measurements of bacterial heterotrophic assimilation rate should be run using these compounds instead of glucose.

     

Hydrocarbon-oxidizing microflora from the water of the Baltic sea and Kurshsky bay polluted after a fuel oil spill

Microbiological studies were conducted in the water of the Baltic sea and the Kurshsky bay polluted with mazut as the result of a tanker wreck in November 1981 as well as in the water of nonpolluted regions. Within the summer of 1982 and 1983, 755 bacterial strains were isolated from water samples taken at three different depths. Bacteria belonging to the genera Pseudomonas, Rhodococcus + Mycobacterium and Arthrobacter predominated in the hydrocarbon-oxidizing cenoses of the Baltic sea and the Kurshsky bay. The central part of the Baltic sea pure from mazut did not differ from the polluted regions in the qualitative composition of the hydrocarbon-oxidizing bacterial flora. Rhodococci and mycobacteria prevailed in the water near harbours, and pseudomonades, in the open waters. The greatest variety of species was found at a depth of 1 m. The proportion between the predominating genera of hydrocarbon-oxidizing bacteria was not stable. The state of studies conducted with the hydrocarbon-oxidizing microflora is analysed and the factors causing discrepancies are discussed. One must keep in mind that it is necessary to use a strictly elective medium and to examine cultures with a microscope many times throughout their growth in the isolation and identification of hydrocarbon-oxidizing bacteria. The interrelationship is analysed between the predominant genera of hydrocarbon-oxidizing bacteria.     

Utilization of different carbon sources by bacteria isolated from the roots of pine seedlings (Pinus silvestris L.) inoculated with root-free, rhizosphere and mycorrhizosphere soil

Studies were carried out on the utilization of different sugars (glucose, fructose, arabinose, xylose, sucrose) and organic acids (acetic, citric, fumaric, propionic, succinic) by fast and slow growing bacteria isolated from the roots of pine seedlings (P. silvestris L.) inoculated with root-free, rhizosphere and mycorrhizosphere soil from nursery and mature pine forest. Sucrose among sugars and propionic acid among organic acids were the less frequently utilized compounds. Sugars were better carbon sources than organic acids. Proportion of isolates utilizing respective sugars or organic acids was, in general, significantly higher among fast growing bacteria as compared with slow growing ones. No significant differences in number of strains assimilating the appropiate sugars depending on their original habitat or kind of soil were observed. Such differences were noted for utilization of some organic acids. 3-factor ANOVA confirmed that the growth speed of bacteria at the moment of their isolation had the strongest effect on utilization of the compounds studied.     

Etude de la fertilité des eaux marines au moyen de tests biologiques effectués avec des cultures d'algues. II. Limitation nutritionnelle et viabilité de l'inoculum

Ten cultures of phytoplankters, including four strains of Skeletonema costatum from different origins, were used to improve some aspects of the bioassay technology. Special attention was paid to the preliminary nutrient limitation of the inocula. When the cells are maintained in nutrient starvation, their carbon, nitrogen, carbohydrates, proteins and above all chlorophyll a contents decrease. The minimum appears after a range of 2–7 days, according to species. The survival of these limited cells and their capacity to give rise to active growing cultures when sub-cultured are different with species, but efficiency in the inocula they provide usually can occur only until the minimum content in cellular components appears. Starved cells and enriched cultures of Chaetoceros lauderi and Skeletonema costatum were used to inoculate several samples of sea water, in order to test the effect of the starvation on the experimental results. It appears that the starved cells increase the sensitivity of the method, but they are more susceptible to substances limiting their growth. In the opinion of the authors the best way would be to use both starved and enriched cells as inocula, but, when this is impossible, cautiously starved cells should be used with unpolluted sea waters.     

Production of oil-processing compounds by microorganisms from the Daqing oil field,China

Twenty pure cultures isolated from formation waters of the Daqing oil field were studied with respect to their capacity to produce surface-active compounds in media with individual hydrocarbons, lower alcohols, and fatty acids. Aerobic saprotrophic bacteria belonging to the genera Bacillus, Brevibacillus, Rhodococcus, Dietzia, Kocuria, Gordonia, Cellulomonas, Clavibacter, Pseudomonas, and Acinetobacter decreased the surface tension of cultivation media from 55-63 to 28-44 mN/m. Strains of Bacillus cereus, Rhodococcus ruber, and Bacillus licheniformis produced biosurfactants most actively. Bacteria of the genera Rhodococcus, Dietzia, Kocuria, and Gordonia produced exopolysaccharides in media with hydrocarbons. Culture liquids of the strains of R. ruber and B. licheniformis exhibited oil-releasing effect. Thus, the Daqing oil field is inhabited by aerobic bacteria capable of producing effective oil-releasing agents.     

Complete genome sequence of Ilyobacter polytropus type strain (CuHbu1)

Ilyobacter polytropus Stieb and Schink 1984 is the type species of the genus Ilyobacter, which belongs to the fusobacterial family Fusobacteriaceae. The species is of interest because its members are able to ferment quite a number of sugars and organic acids. I. polytropus has a broad versatility in using various fermentation pathways. Also, its members do not degrade poly-β-hydroxybutyrate but only the monomeric 3-hydroxybutyrate. This is the first completed genome sequence of a member of the genus Ilyobacter and the second sequence from the family Fusobacteriaceae. The 3,132,314 bp long genome with its 2,934 protein-coding and 108 RNA genes consists of two chromosomes (2 and 1 Mbp long) and one plasmid, and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.     

Temperature affects the response of heterotrophic bacteria and mixotrophic algae to enhanced concentrations of soil extract

To investigate the consequences of increased temperature and enhanced input of dissolved organic matter (DOM) into lakes for heterotrophicic bacteria and for mixotrophic algae which use DOM in addition to photosynthesis, the hypotheses were tested whether (1) both bacteria and mixotrophic algae benefit from increased input of DOM, or (2) increased DOM input enhances bacterial biomass and thereby decreases algal biomass. Growth experiments in batch cultures, exudation measurements, and competition experiments in chemostats were performed at two temperature levels. Increased temperature stimulated the autotrophic growth rate of Chlorella protothecoides. Bacteria and Chlorella increased their heterotrophic growth rates at higher DOM concentration at lower temperature whereas enhanced DOM concentration hardly stimulated their growth at higher temperature. In chemostats, enhanced input of soil extract increased both bacterial and algal biomass at lower temperature whereas bacterial biomass increased only slightly and algal biomass decreased at higher temperature. Thus, the temperature determines the response of microorganisms to enhanced DOM concentration.     

Isolation and characterization of marine oligotrophic bacteria

A significant part of the world ocean is characterized by low absolute nutrients and chlorophyll concentrations. In these oligotrophic environments, bacteria are very abundant and play a vital role in the remineralization of the dissolved organic matter. Bacteria adapted to oligotrophic waters differ from those adapted to richer environments by some genetic and metabolic characteristics. Culture techniques in bacteriology are based on rich media and do not allow the growth of most marine bacteria. New techniques have been developed for the culture of oligotrophic bacteria, which allow to isolate unknown bacteria. Pelagibacter ubique and Sphingopyxis alaskensis belong to these bacteria recently isolated from the marine environment and their study yielded better understanding of how marine bacteria adapt to oligotrophic conditions.     

Etude de la fertilité des eaux marines au moyen de tests biologiques effectués avec des cultures d'algues I. Comparaison des méthodes d'estimation

Several methods of growth potential estimation of marine productivity are discussed: (1) simultaneous measurements of nutrient content and organic production, (2) chemical composition of algal cells, (3) bioassay with algal cultures. This last method appears to be the most convenient; it allows to study the role of a nutrient component both when utilized alone and when combined in a pool of nutrients. Several aspects of bioassay technology are discussed: test species, chemical enrichments and growth estimation of test cultures.     

The estimation of extracellular release by phytoplankton and heterotrophic activity of aquatic bacteria.

The actual extracellular release of organic matter by algae was determined in water samples in which the heterotrophic activity of bacteria was inhibited by gentamycin. Gentamycin rapidly and efficiently inhibited the activity of aquatic bacteria without affecting phytoplankton metabolism. Aquatic bacteria utilized the products of algal extracellular release. The amount of algal photosynthetic products metabolized by bacteria can be taken as a measure of their heterotrophic activity in waters.     

Nutritional Aspects of Dissimilatory Sulfate Reduction in the Human Large Intestine

In contrast to other anaerobic ecosystems, such as marine and estuarine sediments, there is a lack of information on the nutritional requirements of human gut sulfate-reducing bacteria (SRB). Various substrates stimulated sulfate reduction in mixed culture, including short-chain fatty acids and other organic acids, alcohols, and amino acids (but not sugars or aromatic compounds). However, the use of sodium molybdate as a specific inhibitor of sulfate reduction caused an accumulation of ethanol and malonate only, and reduced the rate of utilization of lactate. This indicates the importance of these electron donors for sulfate reduction. Since ethanol and lactate are primarily utilized by members of the Desulfovibrio genus, the results suggest a physiologically important role for this group.  Experiments with two strains of Desulfovibrio desulfuricans isolated from human feces demonstrated that both were able to reduce sulfite, thiosulfate or nitrate in the absence of sulfate. In addition, one strain (DsvUC1) was able to grow by fermentative metabolism, although the second strain (DsvFD1) showed more restricted fermentative growth. The data indicate that desulfovibrios are ecologically the most significant group of SRB in the human colon, and that colonic isolates belonging to this genus are versatile, in terms of both the electron acceptors and donors that they are able to utilize. Received: 24 March 1997 / Accepted: 10 June 1997     

In vitro evaluation of antibacterial substances produced by bacteria isolated from different marine organisms

Bacteria from several groups of marine organisms were isolated and, using direct antibiograms, identified those that produce antibacterial substances, using a human pathogenic strain of Staphylococcus aureus ATCC6538 as revealing microorganism. Bacteria which produce substances that inhibited S. aureus growth were identified through morphological, physiological and biochemical tests. Out of 290 bacteria, 54 (18.6%) inhibited the growth of S. aureus, but only 27 survived for identification. Bivalves, sponges and corals were the most represented from which 41.2, 33.3 and 29.7%, respectively, produced antibacterial substances of the isolated bacteria in each group. The marine species with highest proportions of these bacteria were the hard coral Madracis decactis (62.5%), the sponges Cliona sp. (57.1%) and the octocoral Plexaura flexuosa (50.0%). Out of the 27 strains that produced antibacterial substances, 51.8% were Aeromonas spp. and 14.8% Vibrio spp. Marine bacteria that produce antibacterial substances are abundant, most belong in the Vibrionacea group and were isolated mainly from corals and bivalve mollusks.     

溶藻细菌筛选及溶藻活性物质对铜绿微囊藻生理活性的影响

从太湖水华水体中分离纯化细菌, 再将细菌的LB液体和固体斜面纯培养物分别收集后感染铜绿微囊藻(Microcystis aeruginosa)细胞, 从中筛选出7株具有溶藻活性的细菌, 并对其中一株溶藻细菌THW7的溶藻方式及溶藻活性物质对铜绿微囊藻生理活性的影响进行了初步研究。结果表明: 仅采用细菌的LB液体纯培养物进行溶藻细菌筛选会存在误筛或高估溶藻效率的风险, 而采用细菌的固体斜面纯培养物进行筛选则可避免以上风险; 溶藻细菌THW7通过分泌胞外活性物质的方式间接溶藻; 在THW7无菌滤液胁迫下, 铜绿微囊藻的生长受到显著抑制(P<0.01), 10d溶藻效率可达94.38%, 光合系统活性也显著降低(P<0.01), MDA含量积累, SOD、POD、CAT活性整体呈现先升高后降低的趋势且显著高于对照组(P<0.01)。推测菌株THW7分泌的溶藻活性物质可能作用于铜绿微囊藻细胞的光合系统Ⅱ, 阻碍电子传递, 抑制其光合作用过程, 并对藻细胞产生氧化损伤, 破坏藻细胞细胞膜的完整性, 从而实现溶藻作用。     

Nature and Role of Bacterial Contaminants in Mass Cultures of Thermophilic Chlorella pyrenoidosa

A study was made of bacterial contaminants isolated from an algal mass-culture unit. The study was performed specifically to determine the dependence of the size of bacterial population on algal density and the nature of any association of the contaminants with the algal cell. Growth of the bacterial contaminants on standard medium was also investigated. An estimate was made of the O₂ uptake of the bacterial population under normal operating conditions of the algal massculture system. Viable numbers of bacteria tended to increase with increased algal density. Bacteria were found imbedded in the surface of algal cells when the cultures of algae were characterized by subnormal rates of growth and photosynthetic gas exchange. Bacterial isolates failed to grow in standard medium alone, thus implying a dependency of bacterial growth on material(s) produced by the algae. A slight inhibitory effect on algal growth was noted in the case of two of three of the bacterial isolates. Manometric studies demonstrated that the bacterial population normally found in the algal cultures did not appreciably effect total gas exchange.