The influence of Na+ and K+ on the synthesis and secretion of extracellular glucosyltransferase (GTF; EC 2.4.1.5) and fructosyltransferase (FTF; EC 2.4.1.10) by Streptococcus sanguis NCTC 7865 and Streptococcus sanguis Challis NCTC 7868 has been determined. No FTF and little or no mutansucrase (GTF-I) activities were detectable during growth on glucose or sucrose unless the Na+/K+ ratio of the cultures was kept low. Increasing K+ concentrations stimulated the production of FTF and dextransucrase (GTF-S), but all glycosyltransferase activities decreased in high K+ media when the growth pH was maintained with NaOH instead of KOH, indicating that the Na+/K+ ratio effect was due principally to Na+ inhibition. Significant GTF and FTF activities were detected in a putative GTF- mutant of strain Challis grown in high K+ medium but not in high Na+ medium, suggesting that the mutant might be defective in a regulatory gene.

Hippurate hydrolysis can also be positive with beta-haemolytic streptoccoci groups C and L. The CAMP test which demonstrates the synergistic haemolysis between staphylococcal B-toxin and streptococcal CAMP factor on sheep or ox red blood cells is only positive in about 84% of strains.' Pigment production by group B streptoccoci (GBS), which is orange in colour and has characteristics of a carotenoid,2 was first noted by Lancefield in 1934 in nine of 29 strains grown anaerobically.3 Modifications of media by Fallon,4 Merritt,5 and Islam6 have improved the sensitivity to about 97%.We describe further investigations into this test and modifications to the media described by Islam, and a useful application of one of the modifications. Material and methodsIn the first phase, tests of sensitivity and specificity of the media were done by performing Lancefield

SUMMARYThe concentrations of adenosine 5′-mono-, -di-, and -triphosphate (AMP, ADP and ATP) in sheep digesta were measured by a new bioluminescence method. ATP in rumen digesta varied with time after feeding and diet, and most markedly according to the size of the population of ciliate protozoa: the ATP content of rumen digesta from ciliate-free sheep was less than a quarter of that of faunated animals receiving the same diet. The adenylate energy charge (EC) (ATP + ADP/[ATP + ADP + AMP]), an indicator of metabolic activity, was high (0·77–0·94) in all rumen samples, but did not appear to be as useful a measurement of activity as ATP alone. As digesta passed along the alimentary tract, the ATP content decreased progressively, from 0·9–2·2 μmol/g dry matter in the rumen to 0·5–1·0μmol/g in the abomasum and duodenum, 07middot;03–0·08 μmol/g in ileal digesta and 0·01–0·02 μmol/g in faeces. EC also tended to decrease, further emphasizing the large decrease in microbial activity which occurs in the hindgut.

The fatty acid composition of Streptococcus sanguis NCTC 7865 was not altered by changing the cation composition (Na+/K+) of the growth medium; glucosyltransferase (GTF; EC 2.4.1.5) also remained constant. In contrast, fructosyltransferase (FTF-S; EC 2.4.1.10) production was reduced by at least 50% in medium with a high Na+ concentration. Growth in the presence of ionophores (gramicidin, nigericin or valinomycin) resulted in an increased proportion of saturated fatty acids, principally octadecanoic acid (C18:0), while the proportion of unsaturated fatty acids, predominantly octadecenoic (C18:1) and hexadecenoic (C16:1) acids, decreased. GTF-S production was reduced in the presence of ionophores whereas FTF-S production was completely abolished. Tween 80 significantly increased both GTF-S production and the proportion of unsaturated fatty acids in the cytoplasmic membrane; FTF-S production was unaltered by Tween 80. The production of GTF-S was inversely proportional to the C18:0:C18:1 fatty acid ratio of the cytoplasmic membrane. It was concluded that FTF-S production is directly influenced by protonmotive force (pmf), whereas GTF-S production is affected more by the physical properties of the cytoplasmic membrane, in particular its fatty acid composition. However, as perturbations in pmf generation can lead to variations in membrane fatty acid composition it can be argued that pmf indirectly influences GTF production by changing the saturated:unsaturated or C18:0:C18:1 fatty acid ratio of the cytoplasmic membrane.

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SUMMARYAerosol infection (Al) of Porton outbred mice with Listeria species, exhibiting varying degrees of virulence, was compared with gastric intubation (GI) on the basis of numbers of deaths, 50% lethal dose (LD50) and pattern of listerial infection. The Al route appeared to be more sensitive, efficient and consistent than GI in that it required 105 fewer micro-organisms to obtain infection and death then ensued within 4 days, with GI deaths usually occurring on day 7. All the virulent strains tested caused 100 % mortality by AI, while virulent and avirulent strains were indistinguishable by GI. Bacterial counts in the livers and spleens of infected mice were consistent with the relative virulence of the infectious agent using AI but not in GI mice. There were higher numbers of micro-organisms and more widespread lesions in the organs of AI mice than in GI. Results indicate that Al is an accurate in vivo indicator of virulence in listeria and using Al, bacterial counts in the liver and spleen could replace LD50 tests, thereby reducing the number of animals required for in vivo virulence testing.

Summary. Small numbers of bacteria capable of growing on agar supplemented with amoxycillin 40 mg/L were isolated from the saliva of 9 out of 20 adult volunteers in a previous study. All the bacteria were identified as Streptococcus sanguis although no strains produced dextran in conventional tests. However, using a specific assay, all the antibiotic-resistant strains were found to secrete glucosyltransferases (GTF), the enzymes that synthesise these extracellular polysaccharides ; the production of GTF-S, the enzyme that synthesises dextran, was 2243% less than that of an antibioticsensitive control strain. Enzyme production by both antibiotic-resistant and sensitive bacteria was markedly inhibited by dextran primer. The amoxycillin-resistant bacteria were resistant to other penicillins ; their resistance to erythromycin was variable but they were uniformly sensitive to cephalothin and clindamycin. As dextran production has been proposed as a key factor in the colonisation of damaged heart valves by bacteria such as S . sanguis, these highly resistant bacteria may not pose a threat to the susceptible individual.

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