GERBAM — Deuxième Colloque International de Bactériologie marine — CNRS, Brest, 1-5 octobre 1984
IFREMER, Actes de Colloques, 3, 1986, pp. 273-276
31
A SIMPLE MOST PROBABLE NUMBER METHOD FOR THE ENUMERATION
OF SULPHATE-REDUCING BACTERIA
N. S. BATTERSBY*, A. P. SHARMA** and D. J. STEWART**
** Department of Biological Sciences, Heriot-Watt University Chambers Street,
EDINBURGH EH1 1HX, (U.K.)
* WRC Environment, Henley Road, Medmennam, PO BOX 16 MARLOW, SL7 2HD,
(U.K.)
ABSTRACT - A simple most probable number (MPN) method for the enumeration of sulphate-reducing
bacteria (SRB) in pure and mixed cultures is described. The medium is based on sea or source water and is
reduced by a suspension of Pseudomonas putida cells. This eliminates the need for toxic reducing agents such as
thioglycollic acid which can lower SRB recovery. SRB recovery is comparable to other published methods even
when the MPN medium is deliberately oxidized prior to inoculation.
Key words : sulphate-reducing bacteria, enumeration, Pseudomonas putida
RÉSUMÉ - Une méthode simple basée sur le nombre le plus probable (MPN) est décrite pour l’énumération en
culture pure ou non des bactéries sulfato-réductrices (SRB). Le milieu qui utilise de l’eau de mer ou de source est
ensuite réduit par l’addition d’une suspension de cellules de Pseudomonas putida. Ceci remplace les agents
réducteurs toxiques tel que l’acide glycollique qui peu abaisser la concentration en SRB. La numérotation en
SRB est comparable à celle obtenue par les autres méthodes publiées quand le milieu MPN est oxydé
préalablement à l’inoculation.
Mots clés : bactéries sulfato-réductrices, énumération, Pseudomonas putida
INTRODUCTION
Sulphate-reducing bacteria (SRB) are a physiological-ecological group of obligately
anaerobic organisms which conserve energy by the dissimilatory reduction of sulphate to
sulphide. They are widely distributed in nature and play a major role in the biological
sulphur cycle and in the anaerobic oxidation of organic matter (Jorgensen, 1982). SRB
are particularly active in marine waters where a high concentration of sulphate is
generally found (ca 29 mM).
Their metabolism in such sulphate-rich environments also causes major problems in the
North Sea oil industry. These include :
1) corrosion of pipelines and other metal structures
2) blocking of oil-bearing rock pores.
3) souring of oil and gas.
4) production of toxic hydrogen sulphide gas.
The accurate enumeration of SRB is of prime importance if we are to understand better
their environmental and ecological significance.
273
IFREMER, Actes de Colloques, 3, 1986, pp. 273-276
31
A SIMPLE MOST PROBABLE NUMBER METHOD FOR THE ENUMERATION
OF SULPHATE-REDUCING BACTERIA
N. S. BATTERSBY*, A. P. SHARMA** and D. J. STEWART**
** Department of Biological Sciences, Heriot-Watt University Chambers Street,
EDINBURGH EH1 1HX, (U.K.)
* WRC Environment, Henley Road, Medmennam, PO BOX 16 MARLOW, SL7 2HD,
(U.K.)
ABSTRACT - A simple most probable number (MPN) method for the enumeration of sulphate-reducing
bacteria (SRB) in pure and mixed cultures is described. The medium is based on sea or source water and is
reduced by a suspension of Pseudomonas putida cells. This eliminates the need for toxic reducing agents such as
thioglycollic acid which can lower SRB recovery. SRB recovery is comparable to other published methods even
when the MPN medium is deliberately oxidized prior to inoculation.
Key words : sulphate-reducing bacteria, enumeration, Pseudomonas putida
RÉSUMÉ - Une méthode simple basée sur le nombre le plus probable (MPN) est décrite pour l’énumération en
culture pure ou non des bactéries sulfato-réductrices (SRB). Le milieu qui utilise de l’eau de mer ou de source est
ensuite réduit par l’addition d’une suspension de cellules de Pseudomonas putida. Ceci remplace les agents
réducteurs toxiques tel que l’acide glycollique qui peu abaisser la concentration en SRB. La numérotation en
SRB est comparable à celle obtenue par les autres méthodes publiées quand le milieu MPN est oxydé
préalablement à l’inoculation.
Mots clés : bactéries sulfato-réductrices, énumération, Pseudomonas putida
INTRODUCTION
Sulphate-reducing bacteria (SRB) are a physiological-ecological group of obligately
anaerobic organisms which conserve energy by the dissimilatory reduction of sulphate to
sulphide. They are widely distributed in nature and play a major role in the biological
sulphur cycle and in the anaerobic oxidation of organic matter (Jorgensen, 1982). SRB
are particularly active in marine waters where a high concentration of sulphate is
generally found (ca 29 mM).
Their metabolism in such sulphate-rich environments also causes major problems in the
North Sea oil industry. These include :
1) corrosion of pipelines and other metal structures
2) blocking of oil-bearing rock pores.
3) souring of oil and gas.
4) production of toxic hydrogen sulphide gas.
The accurate enumeration of SRB is of prime importance if we are to understand better
their environmental and ecological significance.
273