Winogradoky, was prepared by filling a large lass cylinder (2,000 ml) about one fourth full with organic rich sulfide containing mud from the

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1 two strategies were employed Sippewisset marsh. The mud was spiked with calciumsulfate as sulfate source. The mud was then covered with seawater till to the top and microscopical observation showed an abundance of Prosthecochloris sealed with parafilm. The column was placed at a north window so as to Starr,M.P. et al.(1981) The Prokaryotes; Volume 1 ; receive adequate ( but not excessive ) sunlight and left to develop for several weeks. A bloom of green sulfur bacteria was observed and After one and a half week green colonies were picked out of the fourth order to enrich for Co oxidizers which are probably not described yet, about one fourth full with organic rich sulfide containing mud from the Winogradoky, was prepared by filling a large lass cylinder (2,000 ml) carbonmonoxide as energy source plus acetate as carbon source (1). In Edition; Prentice Ran. International; Introduction: From the literature it is known that some sulfate reducers Brock,T.D. and Madigan M.T. (1991) Biology of Microorganisms; Sixth Literature: of Frosthecochloris were obtained after a growth period of one week. medium described for phototrophic sulfur bacteria. Three pure cultures Samples were withrawn from the column and added to agar dilution shakes. and fifth agar dilution tubes and transferred anaerobically to a liquid A) Green phdtotrophic bacteria isolated from a Winograsky column Summer Course 1991 B) Carbonmonoxide oxidizing sulfate reducing bacteria Microbial Diversity possess. a carbonmonoxid dehydrogenase and are therefore able to grow on A report of independent projects by Juergen Breitung A Winogradsky column, named for the famous Russian Microbiologist Sergei -t

2 were sealed with black mushroom stoppers. Two bottles serves as a marine sulfate reducer medium using Hungate technique. subsequently approx. 3 g black Sippewisset marsh sediment were added and the bottles 2) filled with 50 ml observation showed that about 70% of the bacteria are vibrios similar the sulfate reducer probe. From this enrichment an agar dilution shake plus 1 % CO in the headspace. After five days of incubation at 30 C tiny probe (described by Gunter) the vibrios showed a positive reaction with shakes were made with marine sulfate reducer medium plus S mm acetate was made in order to get a pure culture of this bacterium. The agar shaped as DesulfovLbrio desulfuri cans. With a 16 srrna fluorescence reducer. In order to test if it is reducible, 1 ml of tube A/S were inoculated in tube A,B,C and D. And again after four days of incubation at 30 C turbidity and H2S was only detected in tube A. Microscopical and a yellow brown color in the test indicating growth of a sulfate Results After four days in tube A/s (from Ron) turbidity could be seen without substrate as a control. plus 1% CC in the headspace and tube D contains 10 ml s.r. medium 10 ml s.r. medium plus 5 mm acetate; tube C contains 10 ml s.r. medium A contains 10 ml marine sulfate reducer medium plus 5 mm acetate plus 1% CO in the headspace (0.17 ml 100% CO/ 17 ml headspace); tube B contains of these enrichments were inoculated (10%) into four hungate tubes: tube of these non acetate oxidizers CO consumption is described (1). Each compound only to acetate ( Desulfov.Lbrio, Desulfomonas etc.). For some transfer). Lactate enriches for sulfate reducers which can oxidize this different course members on lactate were used (second or third enrichment took place in the course. Six sulfate reducer enrichments of 2.) The second enrichment took advantage of the sulfate reducer the course. Also no turbidity could be detected. diagnostic reaction for estimation of formed sulfide handed out during Result: No H2S production could be detected after two weeks using the The bottles were incubated for two weeks on a shaker (80 rpm) at 30 DC. control, two had 2%, two had 5% and two had 20 % CO in the headspace. 160 ml serum bottles were anaerobically (N2/C0 Materials and Methods: 1.) Direct enrichment

3 Conlusion With a four tube test ( CO plus acetate; acetate; CO; % CO in the headspace. This tranfer was made three days before finishing anaerobically in liquid sulfate reducer medium with 5 mm acetate plus 1 the course and since then no growth was detected. 2) 2 ml sample; 20 mm NSA 1) 2 ml sample; 10 mm NSA sulfate. Sulfate was added as indicated. bacterium the following hungate tubes were anaerobically filled with following compounds in 15 ml marine sulfate reducer medium without Materials and Methods: In order to enrich for a NSA utilizing anaerobic might be possible. Direct use by methanogens ( reduction of the methylgroup) and sulfate the methyl group or reduction of the methylgroup with extern electrons) reducing bacteria ( reduction of the sulfite group and/or oxidation of snow. Anaerobic degradation seems to be likely in natural environments. with photochemically produced OH radicals, Virtually nothing is known combination with soil and water contituents when deposited in rain or about the anaerobic biological fate of NSA. It enters into chemical compound. It is generated via atmospheric oxidation of dimethyl sulfide Introduction: NSA is regarded as a major biogenic organic sulfur C) Methane sulfonic acide (MSA) oxidizing or reducing anaerobic bacteria X.iterature: Lupton,F.S. et al. (1984) PENS Microbiology Letters kj/ mol Reaction 2: H H2S Reaction 1: 4CC b 80 kjf mol CO 4 CO following: shaped and it is postulated that a DesulfovLbrio species carries out the source and acetate as carbon source. Most of the organisms were vibrio control)a sulfate reducer was highly enriched, which uses Co as electron yellow brownish colonies were detected and subsequently transferred

4 3) 2 ml sample; 10 mm NSA; 2 mm acetate 4) 2 ml sample; 2 mm acetate 5) 2 ml sample; 10 trim NSA; 20 mm sulfate 6) 2 ml sample; 10 mm NSA; 20 mm sulfate; 2 mm acetate 7) 2 ml sample; 20 mm sulfate; 2 mm acetate 8) no sample; 10 mm NSA as a control 9) 2 ml sample; without substrate as a control Three different samples were used: Sippewisset marsh mud; mud from the above mentioned Winogradaky column; and mud from a little pond at the end of the parking aerea at the bike path. The 27 under anaerobic conditions (N2/C0 2) at 30 C. tubes were incubated Results: After twelve days turbidity could be seen in some tubes. Therefore the tubes were checked for M2S production and methane production in the headspace. Tube 9 showed little fl 2S and methane production depending on the sample taken. So, little methanogenesis and M2S production occurs without added MSA and/or acetate. Only tubes showed significant R25 and methane production comparing with the control tubes:tube 3 and 6 with the Winogradsky column sediment sample. H2S and methane were produced approximately in equimolar amounts. Conclusion: The decribed simultaneous H2S and methane production only with MSA plus acetate results in the following working hypothesis: Acetate is oxidized and the electrons are used to reduce NSA and to methane L C C R3-COO 14 %FpcJ ktitaj CH 11 4.W1S

5 whether it is a interspecies hydrogen transfer could not be resolved and as electron acceptor and is reduced to H2S and methane rather than headepace with an 1.5 bar overpressure. Appropriate controls ( without 3 or 10 mm Fed 3 as Kelly, P.R. and Baker, S.C. (1990) FEKS Microbiology Reviews 87; Literature: beeing oxidized. should be worked out in the future. The results shows that NSA is used Whether it is one organism which carries out the overall reaction or growth was therefore slow. reactions. In the case of tube 7 sulfate has first to be activated and The reduction of the methyl group and of the sulfite group are exergonic 4 CH3 SO CH3 COOH 4 CR Prorsed 94&4tL% HS + 6 Co H20) the bike path were filled to the medium. 99 % methane was given to the swamp and a little freshwater pond at the end of the parking aerea at added for unknown growth factors. About 2 ml mud samples from cedar electron donor were added. Furthermore 0.3 ml 3 % yeast extract was (freshwatermedium without sulfate) and 10 mm KNO anaerobically filled with 40 ml methanogens or sulfate reducer medium oxidizing bacterium several samples were taken. 160 ml serumbottles were Materials and Methods: In order to isolate an anaerobic methane exergonic reaction. However, until now no organism was isolated which is as electron acceptor the methane oxidation to carbon dioxid is a showed a temperature optimum between 25 and 37 0C, indicating an active oxidized methane to carbon dioxide. This strictly anaerobic process microbial participation in this reaction. With nitrate or ferrous iron able to grow anaerobically on methane as energy and/or carbon source. Xntroduction:Anoxic sediments and digested sewage sludge anaerobically D) Enrichment of an anaerobic methane oxidizing bacterium

6 gaschromatograph, could not be detected in any of the bottles. Microbiology ;39; Literature: Zehnder, A.J.S. and Erock, T.D. ( 1980) Environmental miracle, probably it does not exist because of the difficulty of the incubated at 30 C and shaked with 80 rpm. electron acceptors; or without samples) were made. All bottles were or, first oxidation step to methanol. Conlusion: The anaerobic methane oxidizing bacterium remained to be a Results: A decrease of methane, which was measured with a