PLATE VIIIREFERENCES to the isolation and study of Clostridium dzficile are rare. We had done some preliminary investigations when we learned from Dr S. D. Elsden of the Food Research Institute, Norwich, that strains of C. dzficile could deaminate phenylalanine, first to parahydroxyphenylacetic acid (as most clostridia that attack this substrate do), and then decarboxylate the phydroxyphenylacetic acid to p-cresol. None of the other clostridia that we examined were able to perform the latter step.We assumed that an organism that produced p-cresol must necessarily be resistant to it, and we applied this information to the isolation of C. dzficile. MATERIALS AND METHODS Strains.We had altogether 30 strains: three old laboratory strains (nos. G173, 238 and 311), three from the American Type Culture Collection (ATCC nos. 17857, 17858 and 9689), six we had ourselves isolated on cresol-free media from stools of infants (labelled nos. Rl-R6), and 18 that we isolated on cresol-containing medium from human and animal faeces. The following observations are mainly based on our studies of the old laboratory strains (for convenience labelled Nl-N3 and the ATCC strains (labelled N4-NQ. However, the freshly isolated strains have been extensively examined and shown to be identical with these reference strains of C. dificile. Cukivation. Our own early isolates were obtained from the faeces of babies less than 1 week old by plating on Reinforced Clostridial Agar (Oxoid; RCA) adjusted to pH 7-2-7.4, and incubating the cultures in an anaerobic jar for 72 h. Colonies resembling those of C. dificile were then picked off and subcultured in Reinforced Clostridial Medium (Oxoid ; RCM). Later isolates were obtained by lightly emulsifying 0.1 g of faeces, dung, sand or mud in 20 ml of RCM containing 0.2% p-cresol.For the study of the effect of cresols and phenol on growth, a 10 % (v/v) aqueous solution or emulsion of the material to be tested was diluted in RCM to give final concentrations of 0.1, 0.2, 0-3, 0.4 and 0-5 % (v/v). Higher concentrations were made by direct additions of phenol or cresol to RCM, and dissolved by steaming; but as the distribution of cresols in RCM in concentrations of 1 % and over was not Uniform, as shown by the presence of a thin surface film, we abandoned the use of media containing concentrations of phenol greater than 0.5 %.Proteose Peptone water plus 1 % sugar or other substrate, with phenol red as indicator, was used for examination of the biochemical action of C. dificile, cultures being incubated i

Healthcare in developing countries less funded than developed nations (0.8 to 4% vs. 10 to 15%, respectively), and must contend against approximately 1/3 of the population living below the poverty line ($1US/day), poor literacy (58% males/29% females), and less access to potable water and basic sanitation. Cultural and societal constraints combine with these economic obstacles to translate into poor transplantation activity. Donor shortage is a universal problem. Paid donation comprises 50% of all transplants in Pakistan. Post-transplant infections are a major problem in developing countries, with 15% developing tuberculosis, 30% cytomegalovirus, and nearly 50% bacterial infections. The solutions to these problems may seem simplistic: alleviate poverty, educate the general population, and expand the transplant programs in public sector hospitals where commerce is less likely to play a major role. The SIUT model of funding in a community-government partnership has increased the number of transplantations and patient and organ survival substantially. Over the last 15 years, it has operated by complete financial transparency, public audit and accountability. The scheme has proven effective and currently 110 transplants/year are performed, with free after care and immunosuppressive drugs. Confidence has been built in the community, with strong donations of money, equipment and medicines. We believe this model could be sustained in other developing nations.

SUMMARY Saliva was found to be a powerful and specific inhibitor of Neisseria gonorrhoeae. Although 28 other species of bacteria were tested, including Neisseria meningitidis, Neisseria pharyngis var flava, Neisseria lactamica, and Neisseria catarrhalis, we failed to find any others sensitive to saliva under similar conditions. The physical properties of the inhibitory substance indicated that it might be salivary ao-amylase. To test this hypothesis o-amylase was extracted from saliva and was shown to have a high antigonococcal activity. Hog pancreas oc-amylase also showed strong antigonococcal activity, thus the observations indicate that for the strains we tested oc-amylase is inhibitory to gonococci. This observation indicates that either the gonococcal outer cell wall contains some unique lipopolysaccharides or that the gonococcus is unusually dependent on the integrity of these moieties. Whichever speculation proves to be true it indicates a need for a careful study of the gonococcal cell wall.

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The VNTR profile 42235 may represent a family of strains commonly found in communities associated with South Asia.