An outbreak of infection caused by a previously undescribed Gram-negative bacterium affected people attending a hot (37 °C) spring spa in France in 1987. Thirty-five case of pneumonia and two cases of meningitis occurred. None of these patients died. Attack rates were significantly higher for patients above 70 years old and for male patients. An epidemiological comparison of the 26 hospitalized eases with 52 matched controls suggests that spa treatment early on the first day (OR = 4·8) and attendance at the vapour baths (OR = 10·7) were significant risk factors for acquiring the infection. Person-to-person spread was not thought to have occurred. The same bacterium was isolated from the hot spring water. All strains studied shows a single rRNA gene restriction pattern. Epidemiological data indicated that the thermal water was the source of infection. This outbreak stresses the need for increased surveillance of infections in people attending hot spring spas.

Techniques for the fractionation of C′ 1 and C′ 2, and for the specific inactivation of C′ 4 of horse complement are described, and shown to be satisfactory. These three components of complement and also conglutinin are found to be essential to the process of conglutination.

The experiments reported do not exclude the possibility of additional or unidentified fractions of horse serum playing a part in the reaction. Whether or not C′ 3 is essential to the process of conglutination could not be determined from the evidence available.

It was found that by the procedures employed the C′ 4 fraction may be supplied either from the horse complement or from the heated bovine serum added as a source of conglutinin or from both.

When horse complement and bovine antibody against sheep cells are used it is found that conglutination will only result when the complement components are absorbed on to the immune complex in a particular sequence. The sequence is that C′ 1 is absorbed first on to the sensitized cells, and then C′ 2 and C′ 4 are absorbed together. Both these components must be presented together to the sensitized cells carrying C′ 1 if conglutination is to result. Finally, conglutinin acts on the sensitized cells which have absorbed the three complement components, and conglutination results.

The significance of these findings, together with matters of a more general nature, is discussed.

The administration per-orally to mice of the non-absorbable antibiotics polymyxin E, tobramycin and amphotericin B resulted in the elimination of detectable aerobic gram-negative rods from the faecal flora without affecting the total viable aerobic count. The addition of parental cefotaxime to the regime caused a fall in the number of aerobic lactobacilli and an increase in the number of enterococci. The rise was associated with the translocation of viable enterococci to the mesenteric lymph nodes and the spleen. The changes induced by cefotaxime were reversed when the antibiotic was withdrawn. Following withdrawal of all antibiotics the total aerobic faecal flora increased to above normal levels, but there was no associated diarrhoea. Attempts to implant exogenous enterobacteria into the digestive tract resulted in only low level colonization both in treated mice and in control mice. These results may have implications for the use of this antibiotic regime for selective decontamination of the digestive tract in humans, particularly those who are immunocompromised.

1. Human and guinea-pig sperms are susceptible to the most diverse chemicals to almost the same degree.

2. There exists a substance in human semen which protects both human and guinea-pig sperms against certain chemicals, especially sodium oleate, orthovanillin and potassium borotartrate.

3. When perfectly fresh human semen is not available, the spermicidal powers of substances may be tested by using guinea-pig sperms suspended in human semen which has been kept cool.

Much work has already been done in connection with the differentiation of true diphtheria bacilli from diphthomorphic organisms. Some observers have maintained that all disease producing strains could be distinguished from non-pathogenic types by their sugar reactions. Others have laid much stress on some particular forms of virulence test. More recently serological classification has been attempted in order to help to solve this problem.

With regard to sugar reactions Graham-Smith (1908) found that certain strains produced acid in media containing saccharose, while Hine (1913), on the other hand, maintained that no true diphtheria bacillus could ferment this sugar. Eagleton and Baxter (1922) have shown that diphtheria bacilli, virulent and non-virulent, ferment glucose but do not produce any change in saccharose. Jordan and his collaborators (1922) have found that virulent and non-virulent bacilli fermented glucose and maltose and produced no change in saccharose and that their action on dextrin was variable.

Work on the serological classification of these organisms has also shown some considerable variation. Havens (1920) isolated 206 strains of diphtheria bacilli. These represented cultures from acute cases, release cultures and cultures from healthy carriers. He found that these 206 strains could be divided into two serological groups, one group containing 169 strains, the other 37. There was no evidence of any cross agglutination. Durand (1920) isolated 255 strains of typical diptheria bacilli and after excluding those strains which would not form stable suspensions divided them into five groups, A. B. C. D. and E. containing 16, 8, 25, 61 and 40 strains respectively. Eighty-seven strains were found to be inagglutinable by these five sera but nevertheless could absorb agglutinins from such sera. His final figures for the five groups of bacilli are 18, 8, 31, 76 and 51. Durand and Guerin (1921) in a further paper showed that small isolated outbreaks were always due to the same type of bacillus. They also found that healthy carriers gave the same type of bacillus as was found in the cases arising from these sources of infection. Bell (1922) isolated 133 strains of B. diphtheriae indifferently collected from cases and carriers. He was able to agglutinate 80 per cent. of these strains by three monovalent sera. Group I contained 17 strains, Group II 8 strains, Group III 80 strains.

In view of the results which have just been described it has appeared important to investigate further a considerable number of strains as encountered in routine work, not only as regards the serological characteristics of the organisms but also in regard to their morphology, their biochemical reactions, and their virulence for guinea-pigs.

Antibiotic resistance plasmids from organisms that had caused serious epidemics, including those responsible for epidemics of chloramphenicol-resistant typhoid fever and dysentery in Central America, were transferred to a strain of Salmonella typhimurium and of Salmonella gallinarum. The virulence and infectivity of these R+ forms were then compared with the R− parent forms in orally inoculated chickens.

None of the R+ forms were more virulent than their R− parent forms. The mortality rates they produced were either the same as or less than that of their R− parent forms. The mortality rates were not increased by feeding the chickens on diets containing antibiotics against which the plasmids provided resistance.

The removal of the plasmids from some R+ forms of decreased virulence was not accompanied by any alteration in virulence, indicating that they were less virulent mutants of the parent strain that had conjugated preferentially. In other cases their virulence was increased, indicating that the very possession of the plasmid was involved in their decreased virulence. Of four forms of the S. gallinarum strain harbouring the plasmid that had been incriminated in the Central American dysentery outbreak, one was as virulent as the parent R− form and the other three were less virulent. Preferential conjugation by an avirulent mutant was responsible for the lack of virulence of one of them but the very possession of the plasmid appeared responsible for the decreased virulence of the other two. The decreased virulence of de-repressed F+ and I+ forms of the S. typhimurium strain was increased to that of repressed F+ form and of the parent form by plasmid removal.

Organisms of the R+ forms of the S. typhimurium strain were not excreted in larger amounts or for longer periods of time by infected chickens than organisms of the R− parent form were. Neither did organisms of the R+ forms of this strain or the S. gallinarum strain spread more rapidly or more extensively from infected chickens to in-contact chickens than organisms of the R− parent forms did. When antibiotics against which the infecting R+ organisms provided resistance were included in the diet of these chickens the R+ organisms were usually excreted in greater amounts, for longer periods of time and spread more rapidly and more extensively from the infected chickens to the in-contact chickens.

The pH-stability of three strains of rinderpest virus, propagated in tissue cultures, was investigated at 4°C. in veronal-acetate buffers (Michaelis) of pH 3·0–10·7.

An attenuated laboratory strain, in its 95th culture passage (RBOK), showed maximal stability in the pH range 7·2–8·0, the half-life being about 3·7 days. It was relatively stable from pH 4·0 to 10·2, the half-life at the extremes of this range being over 2 hr. At pH 3·0 the infectivity declined very rapidly, the half-life period being 24·0 sec.

One virulent strain (RGK/1) showed a significantly lower resistance than strain RBOK at pH 3/0, probably also at pH 5·0. The inactivation rates for these two strains were, however, not greatly different at pH 10·7. The other virulent strain (RBT/1) was even less stable than RGK/1 at pH 4·0 or 5·0 but of a comparable stability at pH 9·0 and 10·2.

These findings are discussed in comparison with published data on the related viruses of measles and canine distemper. The importance of strain differences in studies of this kind is stressed.

We are grateful to Mr C. S. Rampton, A.I.M.L.T., and Mr L. W. Rowe, F.I.M.L.T., for help in the preparation of the diagrams. This paper is published with the permission of Mr H. R. Binns, C.M.G., O.B.E., Director, E.A.V.R.O.

We followed students in eight elementary schools for rubella antibody from 1993 to 1996 (602 pairs) and 1996–9 (588 pairs) in Gyeonggi Province, Korea. We tested rubella IgG and administered rubella vaccine to the children with the titres <10 IU/ml. The loss rates of rubella IgG during the follow-up periods were 14·3 and 15·8%, respectively. Among vaccinated groups, the loss rate was 18·8%, which was significantly higher than 13·8% of the mixture of natural and vaccine-induced immunity groups. The group that had the lower preceding antibody titre had a higher loss rate of 24·8% compared to 7·2% for the group whose titre was 40 IU/ml or above. In a multivariate analysis, age and gender were not related to antibody loss rate. Under this higher rubella antibody loss rate, in order to prevent congenital rubella syndrome, the immunization for women at childbearing age appears necessary until rubella can be eliminated or controlled.

During 2012 real-time syndromic surveillance formed a key part of the daily public health surveillance for the London Olympic and Paralympic Games. It was vital that these systems were evaluated prior to the Games; in particular what types and scales of incidents could and could not be detected. Different public health scenarios were created covering a range of potential incidents that the Health Protection Agency would require syndromic surveillance to rapidly detect and monitor. For the scenarios considered it is now possible to determine what is likely to be detectable and how incidents are likely to present using the different syndromic systems. Small localized incidents involving food poisoning are most likely to be detected the next day via emergency department surveillance, while a new strain of influenza is more likely to be detected via GP or telephone helpline surveillance, several weeks after the first seed case is introduced.

The prevalence of antibodies to human T-cell lymphotropic virus type I (HTLV-1) was determined in high-risk groups and normal adults in Egypt. Among 647 individuals tested, 6 (0·9%) were confirmed positive by western blot analysis. These included 2 (0·7%) of 279 drug addicts, 1 (3·3%) of 30 patients with sexually transmitted diseases, and 3 (2·2%) of 133 healthy individuals. Antibody was not detected in 47 blood recipients or 158 prostitutes. There was no correlation between sex or geographical location and HTLV-I infection. Fifty-three of the 647 sera (8%) were initially reactive by ELISA, but only 12 sera were repeatedly reactive. Since only 4 of these repeatedly reactive sera were confirmed by the western blot, the frequency of false positives using the DuPont screening ELISA was 1·2% (8/643). Two additional sera, confirmed positive by western blot, had been reactive, but not repeatedly, by ELISA. In comparison to the prevalence of HTLV-I antibody among risk groups in many parts of the world, the prevalence in Egypt was low.

The prevalence of antibodies against Lyme disease spirochaetes in serum samples from 80 forestry workers at high occupational risk of Lyme disease was surveyed by enzyme-linked immunosorbent assay (ELISA) with the OspC-I synthetic peptide. The peptide is part of the outer surface protein C (OspC) amino acid sequence located in the region conserved among Borrelia burgdorferi sensu stricto or sensu lato. Positivity for antibodies against OspC-I was observed in 25 (31·3%) of the forestry workers. Of these positive cases, 12 (15·0%) and 19 (23·8%) were positive for immunoglobulin M (IgM) and IgG antibody, respectively. Among 62 workers who were negative for IgG antibody against B. garinii or B. japonica in our previous study, 9 (14·5%) and 4 (6·5%) were positive for IgM and IgG antibody, respectively, in OspC-I ELISA. These results demonstrate for the first time that Lyme disease in forestry workers can be revealed using OspC-I ELISA. We conclude that forestry workers who show positive results for antibodies against OspC-I have very likely been exposed to Lyme disease spirochaetes, and that those who show positivity for IgM antibody against OspC-I may be in the early stage of Lyme disease.

An investigation in an urban area of south-east Kent of 160 patients who had had either pneumonia or unexplained fever during the period from the end of 1948 to the beginning of 1954 revealed that twenty-three (14%) had probably had Q fever.

A comparison of the epidemiological histories of patients who had had Q fever with those of the remainder showed that the use of a household supply of raw milk was commoner among the former.

Occupational exposure to animals (or animal products), visits to potentially infectious places, or residence near potential sources of infection were not significantly more frequent among the Q-fever cases as compared with the controls.

The isolation of Rickettsia burneti from the bulked milk of some local herds supplying retail dairies in the towns, together with the general epidemiological evidence, suggested that Q fever in the towns was predominantly milk-borne.

1. In the intradermic test virulence of “morphological diphtheria bacilli,” 0·2 c.c. containing 100 million organisms is injected. The control guinea-pig receives 500 units of antitoxin the night prior to the test. The test animal receives 125 units six hours after the injection of the cultures. A known virulent organism is always injected as a control; 12 to 20 cultures can be tested on one guinea-pig.

Of 2368 swabs received, 1388 (59 per cent.) showed morphological C. diphtheriae. Fromthese 1286 pure cultures were isolated (92·7 per cent.). In 234 positive swabs from cases of clinical diphtheria, in a series examined under very favourable conditions, morphological C. diphtheriae were obtained in 231 (98·7 per cent.) and of these 229 (99·1 per cent.) were virulent. In 758 positive cultures from carriers morphological C. diphtheriae were isolated in 715 (94 per cent.); 50 per cent. were virulent C. diphtheriae, 39 per cent. avirulent, 3·6 per cent. of Hofmann group, 0·3 per cent. Xerosis and 6.4 per cent. unclassified diphtheroids.

3. Only one culture of intermediate virulence was found, i.e. uncertain reactions occurred with a dose of 100 million organisms while clear reactions occurred with 1000 million.

4. Out of 626 positive cultures from cases of diphtheria, both virulent and avirulent bacilli were isolated in two instances, once from the throat and once from the nose.

5. A monovalent antitoxin (Park Williams 8) nenutralised all of 819 virulent strains.

6. Amongst 39 cultures obtained from the ear in post-diphtherial conditions, virulent C. diphtheriae were isolated fourteen times.

7. Impure primary (“field”) cultures gave reliable results (94 per cent.) when more than one-third of the organisms seen in the smear made from the overnight culture were morphologically C. diphtheriae. If fewer C. diphtheriae were present in the smear th e primary culture gave uncertain results.

The prevalence of specific HSV-2 antibodies was studied in Tanzanian and Norwegian sexually transmitted disease (STD) patients (1095) and non-STD patients (488). Correlates to demographic and behavioural factors were evaluated. Seropositivity was determined by the non-commercial peptide-55 enzyme-linked immunoassay. The prevalence of HSV-2 antibodies was 70% in Tanzanian and 17% in Norwegian STD patients, 35% in Tanzanian blood donors and pregnant women, and 4, 7 and 14% in Norwegian medical students, blood donors and pregnant women respectively. A higher HSV-2 prevalence was associated with female sex, increasing age, previous STDs, history of genital HSV infection, coitarchal age (age at first intercourse) <15 years and HIV seropositivity. Compared to previous data, the prevalence of HSV-2 antibodies in Tanzanian STD patients has increased remarkably. In Norwegian STD patients our results are consistent with, or lower than, the prevalence previously reported in Western Europe. Demographic rather than behavioural factors were associated with higher prevalence of HSV-2 antibodies in STD patients.

1. The six ‘viruses’, Liverpool, Danysz, London, Ready Rat Relief, Institut Pasteur and Ratin, which are the principal bacterial cultures at present employed for anti-rodent control in Great Britain, have been examined. 2. By means of reciprocal absorption tests all these six strains were found to be serologically identical with S. enteritidis Gaertner, antigenic structure, IX:gom:—. 3. From the results of the fermentation tests, which may be used to subdivide this serological type, Liverpool, Danysz, Ready Rat Relief and Ratin were assigned to the var. danysz subgroup; while the London and Institut Pasteur strains could not be distinguished from the classic S. enteritidis type. 4. Both of these subgroups are pathogenic for man, and evidence is cited which shows quite clearly that human cases of gastro-enteritis have been caused by the use of virus preparations. There are, also, reasonable grounds for believing that these bacterial types may be pathogenic for a number of domestic animals, including some poultry.

1. 1000 urinary and 500 faecal cultures of coliform bacteria were studied by means of the methyl-red, citrate and indol tests and also by their fermentation reactions upon saccharose and dulcite.

2. 8 per cent, of the faecal strains and 52 per cent, of the urinary strains were of the aerogenes type, i.e. methyl-red negative and citrate positive. Indol was produced in 94 per cent, and 43 per cent, of cases respectively.

3. This would suggest additional evidence that the coli group are quickly outnumbered by the aerogenes group once they leave the faeces, and also that the presence of the aerogenes type is not confined even in large numbers to an extra-corporeal habitat.

4. In water bacteriology the constancy or change in the relative proportions of coli and aerogenes strains present is the goal at which to aim. The aerogenes variety cannot of itself be regarded as non-excretal or even non-faecal.

5. From a sanitary standpoint, the classification of coliform bacteria by means of the methyl-red and citrate tests cannot be regarded as entirely free from error so long as (a) the aerogenes strains are found in faeces, in however small numbers, (b) these strains occur as the predominant type in urine, and (c) intermediate strains are found in the faeces, urine, soil and water.

6. The principal value of these tests lies in their comparative specificity in identifying B. coli of immediate faecal origin. The presence of such an organism, interpreted together with the indol test, may be said to suggest dangerous faecal pollution if found prominently in a water sample. The presence of other types identified by the same tests cannot be regarded as having the same significant value in regard to a negative opinion.

7. A plea is made for a further study of coliform bacteria by means of the tests described in relation to the biological behaviour of the bacteria under different environmental conditions.

Selenite F broth prepared in the laboratory from single ingredients was found to be significantly more efficient than nutrient broth in the isolation of Shigella sonnei from human faeces. It was more efficient than direct plating on deoxycholate citrate agar for diagnosing Sonne dysentery in a local outbreak. The difference was significant. There was, however, no significant advantage for enrichment over direct plating when stools posted from Bristol to Cardiff were examined.

Laboratory prepared selenite F broth was preferable to the same medium purchased commercially from two different firms. The product of one firm was significantly better than that of the other.

Two methods of sterilization of laboratory prepared selenite F broth were studied. Seitz filtered selenite was less inhibitory to the growth of small numbers of Shigella sonnei. Sterilization by heating in free steam for periods of 15 mm and over impaired the ability of selenite F broth to allow multiplication of small numbers of Shigella sonnei.

A high percentage of non-typable (NT) Staphylococcus aureus strains was isolated in Spanish hospitals during 1984 and 1985. Several alternative methods of typing were employed to study these isolates. These were: phage-typing at 1000 × RTD, phage-typing after heat-treatment (48 °C), thermal shock (56 °C), reverse-typing and induction of additional phages. Using these methods the number of NT isolates was reducedby 60%. Best results were obtained with heat-treatment. Additional phages and reverse-typing were also useful.

A scheme for the study of outbreaks and sporadic cases caused by NT strains is proposed using the methods described.

A peptide library was used to screen for regions containing potential linear B-cell epitope sites in the glycoproteins and nucleoprotein of Crimean-Congo haemorrhagic fever virus (CCHFV) in an enzyme-linked immunosorbent assay (ELISA). The library consisted of 156 peptides, spanning the nucleoprotein and mature GN and GC proteins in a 19-mer with 9-mer overlap format. Using pooled serum samples from convalescent patients to screen the library, six peptides were identified as potential epitope sites. Further testing of these six peptides with individual patient sera identified two of these peptides as probable epitope sites, with peptide G1451–1469 reacting to 13/15 and peptide G1613–1631 to 14/15 human sera. These peptides are situated on the GC protein at amino acid positions 1451–1469 (relative to CCHFV isolate SPU103/97) (TCTGCYACSSGISCKVRIH) and 1613–1631 (FMFGWRILFCFKCCRRTRG). Identified peptides may have application in ELISA for diagnostic or serosurveillance purposes.

A total of 1799 swine sera collected in Toyama prefecture in the central part of Japan during the years 1978–82 were tested for antibody against swine influenza virus (SIV), A/New Jersey/8/76 (H1N1). A high prevalence of antibody was observed in the years after the severe epizootic of SI, 34·5% in 1979 and 51·7% in 1982. In other years, the percentages of positive sera were low and ranged from 1·7 to 12·4%. Regional variations were seen in relation to a small scale epizootic. No antibody to SIV was detected in any of the sera collected during the warm season. In the following dry and cold winter, however, a severe epizootic occurred among the swine populations.