Dissimilatory Fe(III) and Mn(IV) reduction.

A dissimilatory Fe(III)- and Mn(IV)-reducing microorganism was isolated from freshwater sediments of the Potomac River, Maryland. The isolate, designated GS-15, grew in defined anaerobic medium with acetate as the sole electron donor and Fe(III), Mn(IV), or nitrate as the sole electron acceptor. GS-15 oxidized acetate to carbon dioxide with the concomitant reduction of amorphic Fe(III) oxide to magnetite (Fe(3)O(4)). When Fe(III) citrate replaced amorphic Fe(III) oxide as the electron acceptor, GS-15 grew faster and reduced all of the added Fe(III) to Fe(II). GS-15 reduced a natural amorphic Fe(III) oxide but did not significantly reduce highly crystalline Fe(III) forms. Fe(III) was reduced optimally at pH 6.7 to 7 and at 30 to 35 degrees C. Ethanol, butyrate, and propionate could also serve as electron donors for Fe(III) reduction. A variety of other organic compounds and hydrogen could not. MnO(2) was completely reduced to Mn(II), which precipitated as rhodochrosite (MnCO(3)). Nitrate was reduced to ammonia. Oxygen could not serve as an electron acceptor, and it inhibited growth with the other electron acceptors. This is the first demonstration that microorganisms can completely oxidize organic compounds with Fe(III) or Mn(IV) as the sole electron acceptor and that oxidation of organic matter coupled to dissimilatory Fe(III) or Mn(IV) reduction can yield energy for microbial growth. GS-15 provides a model for how enzymatically catalyzed reactions can be quantitatively significant mechanisms for the reduction of iron and manganese in anaerobic environments.

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Novel mode of microbial energy metabolism: organic carbon oxidation coupled to dissimilatory reduction of iron or manganese.

The sensitivity of the requirement of Methanobacterium ruminantium strain M1 to a new coenzyme, 2-mercaptoethanesulfonic acid (HS-CoM) was examined by use of new techniques that were developed for rapid and efficient handling of large numbers of cultures of methanogenic bacteria. The system uses sealed tubes that contain a gas mixture of 80% hydrogen and 20% carbon dioxide under a pressure of 2 to 3 atm. This modification of the Hungate technique reduces variability among replicate cultures and simplifies the dispensing, sterilization, and storage of liquid media as well as the transfer and maintenance of methanogenic bacteria. Results indicate a limit of sensitivity of the assay at 5 nM HS-CoM, with half-maximal growth at 25 nM HS-CoM. Coenzyme activity could be replaced by 2,2'-dithiodiethanesulfonic acid at a half-molar equivalent of the HS-CoM concentration, or by 2-(methylthio)ethanesulfonic acid on an equimolar basis. These data reveal a very sensitive and precise requirement for HS-CoM in the nutrition of this fastidious anaerobe.

New approach to the cultivation of methanogenic bacteria: 2-mercaptoethanesulfonic acid (HS-CoM)-dependent growth of Methanobacterium ruminantium in a pressureized atmosphere.

This chapter will deal mainly with the characteristics of bacteria from the rumen that have been successfully cultivated in the laboratory. For some ecosystems, particularly those dominated by slow-growing or specialized microorganisms, it has become clear that only a very small fraction (often <1%) of the total microbial diversity has been recovered by cultural methods (Amann et al., 1995) and that descriptions of the ecosystem based on the available isolated strains can be highly misleading. These discrepancies are apparent both from comparison of direct microscopic and culturable counts, and from direct analyses of ribosomal RNA sequence diversity. In the rumen, organisms surviving in significant numbers must have growth rates sufficient to counteract the constant dilution due to turnover of rumen contents, and there are indications that the discrepancies may be less extreme. Leedle et al (1982) found that the culturable count fluctuated with time after feeding between 14% and 74% of the direct microscopic count in the rumens of animals fed on two different diets. Since the viability of several rumen species is known to change upon starvation, the lower figure could partly reflect changes in the viability of known, culturable species. Thus it is to be hoped that the major obstacles to cultivation of the most numerous rumen bacteria have been overcome by the development of sufficiently rigorous anaerobic methods and of suitable isolation media. It remains likely, however, that some functionally important groups (e.g. obligate syntrophs) may not have been recovered; Mclnerney et al (1981) used co-culture with Desulfovibrio in the presence of sulphate to isolate a fatty acid-oxidizing bacterium similar to Syntrophomonas wolfei from bovine rumen contents.

The rumen bacteria

Biological methanogenesis plays a major role in the carbon cycle on Earth. Methanogenesis is the terminal step in carbon flow in many anaerobic habitats, including marine and freshwater sediments, marshes and swamps, flooded soils, bogs, geothermal habitats, and animal gastrointestinal tracts. CH4 escaping from anaerobic habitats can serve as a carbon and energy source for aerobic methanotrophic bacteria, and can escape to the atmosphere, where it is a major participant in atmospheric chemical reactions and is an important greenhouse gas.

Physiological Ecology of Methanogens

Habitats devoid of oxygen include the interior of the alimentary tracts of most mammals, the lower portions of many oligotrophic lakes, the sediment underlying bodies of water, and water logged soils. Water, the continuous phase in all these habitats, is chiefly responsible for the lack of O2. One ml of water equilibrated with air contains only about 8 /il of O2, compared to 210 ?l 02/ml of air. This oxygen is soon used by aerobic microbes if other suitable foods are available. Oxygen is re moved by metabolism as rapidly as it enters anaerobic habitats. Both euryoxic and anaerobic bacteria have evolved in these habitats. In most continuously anaerobic habitats obligate anaerobes are more a bundant than euryoxic types, possibly because the latter bear a burden of aerobic metabolic capacities unused in the anaerobic environment. Usual aerobic petri plates or similar containers are suitable to culture the euryoxic microbes, but most anaerobes fail to grow in the presence of air. The anaerobes can be classed as oxyduric, i.e. surviving exposure to O2 but not growing in its presence, and oxylabile Species, killed by exposure to O2. Many oxyduric anaerobes can be handled in much the same fashion as aerobes, except that after plates are streaked they must be incubated

https://www.jstor.org/stable/info/20111550

J. Macy

Papers

The Roll-Tube Method for Cultivation of Strict Anaerobes

The roll tube method for cultivation of strict anaerobes