THE STUDY OF OBLIGATELY ANAEROBIC BACTERIA iii„-21 



SEROLOGICAL REACTIONS 



The serological relationships of the spore-forming anaerobes 

 have been reviewed (McCoy and McClung, 1938) and it is sug- 

 gested that this paper should be consulted as a background and for 

 further references by those who are interested in this topic. The 

 toxin-antitoxin reaction is of value as a taxonomic aid with certain 

 species. In such an instance one takes advantage of the fact that 

 relationships may be established by the success or failure of the re- 

 action of antitoxin, prepared against the toxin of a known organism, 

 with the toxin from the unidentified strain. In some instances the 

 anaerobic species are monotypic with respect to toxin formation. In 

 other species this is not true and subgroups have been established 

 within these species or species groups on the basis of non-cross 

 neutralization tests. 



The problem of toxin production may be briefly mentioned. Although studies 

 have been initiated on the possibilities of synthetic media for this purpose, such studies 

 are designed to provide toxin for chemical purification investigations and for produc- 

 tion of toxoid. If it is desired to test for the possibility of production of toxin by a 

 particular culture, it is unnecessary to use a synthetic medium since one of the complex 

 media will serve as well and because less diflSculty with regard to growth is encountered. 

 For organisms producing the tetanus or botulinus toxin use the beef heart infusion. 

 For the gangrene group use the same medium or glucose meat infusion or the medium 

 of Reed, Orr and Baker (1939). For formulae consult Leaflet II. Use the Berkefeld 

 or Mandler filter to remove cells from the liquid of a 24-72 hour culture. Discard 

 the first 25 ml. of filtrate before collecting the test sample. 



For the agglutination reaction, cells for antigen suspensions may be prepared by 

 centrifuging from broth cultures in which maximum growth is attained quickly. For 

 the pathogenic group ^ucose meat infusion broth or perhaps thioglycollate broth 

 should be used. For the butyric-butyl group, one should employ 1% tryptone broth 

 or yeast infusion broth with 0.5 to 1.0% glucose, with a heavy inoculation from a liver 

 broth culture into deep tubes or bottles of the medium chosen. Care should be taken 

 to collect the cells before excessive slime formation is evident in order to produce a 

 stable antigen. 



REFERENCES 



Barker, H. A. 1936. Studies upon the methane-producing bacteria. Arch. Mikrob., 

 7, 420-438. 



Bergey, D. H., Breed, R. S., Murray, E. G. D., and Kitchens, A. P. 1939. Bergey's 

 Manual of Determinative Bacteriology, 5th Ed. Williams and Wilkins, Balti- 

 more. 1032 pp. 



Brewer, J. H. 1942. A new petri dish cover and technique for use in the cultiva- 

 tion of anaerobes and microaerophiles. Science, 95, 587. 



Brown, J. H., and Brewer, J. H. 1938. A method for utilizing illuminating gas 

 in the Brown, Fildes, and Mcintosh or other anaerobe jars of the Laidlaw princi- 

 ple. J. Lab. and Clin. Med.. 23, 870-874. 



Committee Upon Anaerobic Bacteria and Infections. 1919. Report on the anaerobic 

 infection of wounds and the bacteriological and serological problems arising there- 

 from. (Gt. Brit.) Med. Research Council, Spec. Rpt. Ser., 39, 1-182. 



Dack, G. M. 1940. Non-spore-forming anaerobic bacteria of medical importance. 

 Bact. Rev., 4, 227-259. 



Fildes, P. 1931. Anaerobic cultivation. Chap. VI in System of Bacteriology, Vol.9, 

 (Gt. Brit.) Med. Research Council. 



