Recent epidemiological studies have shown that death-rates from certain chronic diseases are higher in areas with soft than in areas with hard drinking-water. In the striking negative correlation found in the county boroughs of England and Wales between cardiovascular mortality and water hardness the important underlying factor is apparently the water calcium. Interest is therefore focused on the dietary significance of calcium present in drinking-water. In relation to that interest, the present report gives a quantitative account of the composition of tea infusions prepared with waters containing different amounts of calcium. It is shown that a substantial part of water calcium is taken up by the tea leaf during the preparation of infusions. The analysis of the infusions covers a wide range of individual components, including trace metals and polyphenolic substances. It appears that the principal change caused in infusion composition by the presence of calcium in the water is a substantial reduction in the relatively high oxalate content. The question is raised whether there may be some connexion between the ‘ water factor’ in cardiovascular disease and the absorption of oxalates from foods.

The hygiene and marketing of fresh cream in England and Wales was investigated by a working party of the Public Health Laboratory Service (PHLS) between 1 October 1968 and 31 July 1969. Thirty-one cream-producing dairies were visited and observations made in the light of the Code of Practice published by the Milk and Milk Products Technical Advisory Committee of the Ministry of Agriculture, Fisheries and Food, and the Scottish Home and Health Department. Suggestions are made in this report to strengthen the code.

A total of 5184 samples of fresh cream comprising 4385 heat-treated, 282 clotted and 517 untreated were examined. Details of production, age of the cream, distance and mode of distribution along the retail chain were available for most samples, and this history was, as far as possible, related to the laboratory findings. In the laboratory the samples were examined by the methylene blue test, colony count, conform and Escherichia coli I test and for pathogens. Staphylococcus aureus was grown from 59 out of 3417 samples of cream; 54 of these were from untreated cream. Phage typing indicated that a proportion of these strains were of animal, presumably bovine, origin. Other human pathogens isolated included one each of Salmonella typhimurium, Brucella abortus, E. coli type O126 and Clostridium welchii.

It appeared that heat-treated cream was much better, bacteriologically, than untreated; and that large dairies, in general, had better premises, more hygienic methods of preparation and the advantages of mechanical filling and capping of cartons; thus they offered the consumer a better product, bacteriologically, than some of the smaller dairies.

A statistical analysis of the results of the tests used showed the methylene blue test, in spite of some anomalies, to be of more use as a screening test than any of the others. As this test is cheap and easy to carry out, the working party thought that it should be the test of choice. In view of the known anomalies, however, the working party recommends that the test should remain a screening or advisory test and have no penal function. Results should be reported as follows: cream not decolorizing the dye in 4 hr. at 37° C. (± 0·5°) after overnight incubation at 20° C. (± 0·5°) would be accepted as satisfactory; between ½ and 4 hr. at 37° C. (± 0·5°) as fairly satisfactory; but if the dye were decolorized at the end of the overnight incubation the sample would be provisionally classified as ‘unsatisfactory; requires further investigation’. A repeat sample would then be examined. If samples were repeatedly unsatisfactory, consultations would be arranged between the local authority, the laboratory and the dairy, with perhaps inspection of the dairy premises and retail storage conditions and further, more searching tests on samples made in an effort to eradicate production faults.

The second patient with Arizona gastroenteritis to be diagnosed in Britain had just returned from the United States, where his illness began.

Strontium chloride B medium and E.E. broth have been found effective in the recovery of Edwardsiella, Salmonella and Arizona species from the cloacal contents of tiger snakes (Notechis scutatus). Strontium chloride B medium was superior to E.E. broth.

At least one bacterial species was detected in each of the 60 reptiles examined, and all three organisms were recovered from each of 29 snakes on a single examination.

Strontium chloride M, strontium selenite and Rappaport enrichment media and bismuth sulphite agar, although satisfactory for the isolation of Salmonella and Arizona species, were found unsuitable for Edwardsiella tarda.

Ten strains of myocoplasmas were recovered from cultures of synovium or cultures inoculated with synovial fragments from rheumatoid arthritis and one from osteo-arthritis. The source of the organisms is not known. Patients with rheumatoid arthritis had no complement-fixing antibody and no fluorescent staining antibody against the mycoplasmas isolated and no mycoplasma antigen was detected by immunofluorescence in sections of synovia and in synovial fluids.

The strains isolated were of two main serological types and could be distinguished by direct fluorescent antibody staining from standard types of human commensals and the common tissue-culture contaminants. One may be Mycoplasma laidlawii.

1. The European rabbit flea Spilopsyllus cuniculi (Dale) bred successfully in wild rabbits on three properties in New South Wales and, within two breeding seasons, almost every rabbit shot within a quarter of a mile of a release site was infested.

2. It was demonstrated that the flea transmitted myxoma virus in the field.

3. In areas where more than 75 % of the rabbits shot at the beginning of the breeding season were flea-infested and myxoma virus was present, populations failed to show the expected summer build-up.

Studies were made in a modified hospital ward containing 19 beds, 14 of them in the open ward, one in a window-ventilated side-room, two in rooms with partial-recirculation ventilators giving 7–10 air changes per hour, and two in self-contained isolation suites with plenum ventilation (20 air changes per hour), ultra-violet (UV) barriers at doorways and airlocks.

Preliminary tests with aerosols of tracer bacteria showed that few bacteria entered the plenum or recirculation-ventilated rooms. Bacteria released inside mechanically ventilated cubicles escaped into the corridor, but this transfer was reduced by the presence of an airlock. UV barriers at the entrance to the airlock and the cubicle reduced the transfer of bacteria from cubicle to corridor.

During a period of 4 years while the ward was in use for surgical and gynaecological patients, the incidence of post-operative sepsis and colonization of wounds by multiple-resistant Staphylococcus aureus was lower (though not significantly lower) in the plenum-ventilated rooms than in the open ward, the recirculator-ventilated cubicles and the window-ventilated cubicles. Nasal acquisition of multiple-resistant Staph. aureus was significantly less common in the plenum-ventilated than in the recirculator-ventilated cubicles and in the other areas. Mean counts of bacteria on settle-plates were significantly lower in the plenum-ventilated cubicles than in the other areas; mean settle-plate counts in the recirculator-ventilated cubicles were significantly lower than in the open ward and in the window-ventilated side-room; similar results were shown by slit-sampling of air. Mean settle-plate counts were significantly lower in all areas when the ward was occupied by female patients. Staph. aureus was rarely carried by air from plenum-ventilated or other cubicles to the open ward, or from the open ward to the cubicles; though staphylococci were transferred from one floor area to another, they did not appear to be redispersed into the air in sufficient numbers to infect the patients. Ultra-violet irradiation caused a significant reduction in the total and staphylococcal counts from the floors of airlocks, and a significant reduction of total counts in the air.

Pyocine typing was used for the characterization of 134 Pseudomonas aeruginosa strains isolated from bovine mastitis. The scheme of Gillies & Govan (1966) was adopted with some modifications, and the procedure gave 89·6% typability. Pyocine type 1 strains were most commonly encountered and were followed in frequency by types 10 and 3. The introduction of two additional indicator strains allowed for division of these types into subtypes.

In spite of some limitations, discussed in the paper, the pyocine typing scheme proved to be useful in ‘marking’ P. aeruginosa strains and in following their association with bovine mastitis in various herds.

Viable counts of beta-haemolytic streptococci per ml. of saliva were made in the following groups: (1) children with acute streptococcal sore throat, (2) children with acute non-streptococcal sore throat, (3) children who had no sore throat but were streptococcal throat carriers, (4) children who neither had a sore throat nor were streptococcal throat carriers.

The mean counts from cases of streptococcal sore throat and from streptococcal carriers were respectively 1·4 × 106 and 2·5 × 105 per ml.

In a comparison of the efficiency of the throat swab, sublingual swab and specimen of saliva in isolating beta-haemolytic streptococci from the upper respiratory tract, culture of saliva produced the best results.

The construction of a Regional Burns Centre in Pinderfields General Hospital, Wakefield, presented an opportunity to study Pseudomonas aeruginosa infection in patients with extensive burns. During the first year (Barclay & Dexter, 1968) a system of disinfection and bacteriological control created conditions permitting more detailed studies to be undertaken which resulted in a significant reduction of infection and cross-infection.

The use of plastic isolators and of an ‘air curtain’ isolator for protection of patients against infection was studied in a burns unit.

Preliminary bacteriological tests showed that very few airborne bacteria gained access to a plastic ventilated isolator; even when the filter and pre-filter were removed from the air inflow, settle-plate counts inside the isolator were much lower than those in the open ward, but the difference was smaller in tests made with an Anderson air sampler, which showed also that fewer large bacteria-carrying particles appeared inside the isolator than outside it. An open-topped isolator allowed virtually free access of bacteria from ambient air. The numbers of airborne bacteria inside an air curtain were appreciably lower than the counts of airborne bacteria in the open ward, but not as low as those in the plastic ventilated isolator.

Controlled trials of isolators were made on patients with fresh burns of 4–30 % of the body surface; the patients were given no topical chemoprophylaxis against Staphylococcus aureus or Gram-negative bacilli. Patients treated in plastic isolators showed a significantly lower incidence of infection with Pseudomonas aeruginosa than those treated in the open ward; this protective effect was shown by isolators with or without filters or with an open top. Ventilated isolators, which protected patients against personal contact and airborne infection, gave a limited protection against multi-resistant ‘hospital’ strains of Staph. aureus, but no such protection was given by an open-topped isolator, which protected only against personal contact infection, or by air curtains, which protected only against airborne infection; the air curtain gave no protection against Ps. aeruginosa, and there was no evidence of protection by any isolator against Proteus spp. and coliform bacilli.

Both the controlled trials and evidence from the bacteriology of air, hands, fomites and rectal and nasal swabs taken on admission and later, supported the view that Ps. aeruginosa is transferred mainly by personal contact, Staph. aureus probably by air as well as by contact and coliform bacilli mainly by self infection with faecal flora, many of which are first acquired from the hospital environment in food or on fomites.

The use of plastic isolators is cumbersome, and of limited value except in the control of infection with Ps. aeruginosa. For this reason and because of the effectiveness of topical chemoprophylaxis such isolators are unlikely to have more than an occasional use in the treatment of burns. Though air curtains greatly reduce airborne contamination, their use in a burns unit does not appear to protect patients against infection when the alternative (and, for Ps. aeruginosa, more important) routes of contamination by personal contact and fomites are left open.

This paper reports a study carried out to clarify the mechanisms involved in adsorption of influenza A and B viruses on iron oxide. Accordingly, the amounts of virus that are adsorbed from virus suspensions of varying concentrations per unit surface area of magnetic or non-magnetic oxide at fixed temperature and time have been determined. The principles involved are clearly the same as those involved in multiple equilibria during the interaction of particles with a large number of combining sites with different intrinsic affinity. Consequently, the amount of virus that is adsorbed per unit mass of iron oxide depends on the size of the adsorbent area, not on its magnetic property. Owing to a significant difference between the affinities of influenza A and B particles for the binding sites on iron oxide, unit surface area of the adsorbent is invariably capable of adsorbing significantly greater amounts of influenza A than B particles. The practical implications of these findings are that a better understanding of the mechanisms involved in virus adsorption on iron oxide will permit a more efficient separation of virus particles from impurities. The simplicity and the rapidity of the technique and the cheapness of the equipment required suggest that the iron oxide method is of great value for both small- or large-scale viral purification, whether it is used as a single step procedure or as a primary step followed by zonal separation.

No evidence of vaccine virus transmission was found in two studies where Wistar RA 27/3 rubella vaccine was administered intranasally. Vaccine was immunogenic in all of 23 vaccinated children in one study, while in the other only 5 of the 11 vaccinees developed antibody. The reduced seroconversion rate in the latter study appears to have been caused by one or a combination of factors, including the vaccination technique, the presence of infective nasal conditions in vaccinees and the titre of vaccine used.

Bacteriological monitoring of penicillin therapy in 30 children with streptococcal sore throats was performed by means of salivary and throat-swab culture, on the first, third, fifth and ninth days after therapy had started. Counts of beta-haemolytic streptococci per ml. of saliva were also performed.

Results showed that salivary culture and estimation of the numbers of beta-haemolytic streptococci in the saliva were much more sensitive indices of the effectiveness of penicillin treatment than throat-swab culture.

Dissociating chemical treatments employing surfactant-solvent systems were applied to purified influenza A and B viruses to obtain viral preparations possessing a significantly higher or lower haemagglutinating activity than the intact virus. All preparations, whether with high or low haemagglutinating activity, with the exception of envelope protein solubilized by Triton X-100, were significantly lacking in the ability to excite the formation of haemagglutination-inhibiting and virus-neutralizing antibodies in inoculated ferrets. In contrast to other treatments, Triton X-100 treatment of virus significantly enhanced the antigenicity of viral protein as judged by virus neutralization and haemaggmtination inhibition tests. Yet the haemagglutinating activity of the envelope protein solubilized with Triton X-100 was about 1 % that of the intact virus. Results suggest that the correlation assumed to exist between the haemagglutinating activity of influenza virus and its ability to excite the formation of humoral antibodies is coincidental. Another important point is that the specific antigenicity of viral protein may be lost or enhanced owing to effects, other than solubilization, by surface-active agents.

Thirteen men wintering on an Antarctic base were isolated from other human contact for 10 months. During this period Coxsackievirus A21 and later influenza A2 virus were administered to some of the men. Serum samples were collected from each of the men at monthly intervals.

Coxsackievirus A21 produced symptoms and apparently spread to uninoculated men. It also appears that repeated re-infections occurred and that the virus persisted in this small community for most of the period of isolation. HI antibody responses in the absence of neutralizing antibody responses seem to be transient.

The vaccine strain of influenza virus induced antibody responses but did not cause symptoms. There was no evidence of spread to uninoculated men.

Antibody titres against influenza C, parainfluenzaviruses 1 and 2 and coronavirus OC43 did not fall significantly during isolation.

An outbreak of respiratory illness occurred at the end of isolation and its origin was traced. No causative agent was detected.

The three heterologous anti-sera currently provided for tetanus prophylaxis have been compared with reference to the production of untoward reactions in 498 patients, and to the blood antitoxin concentrations produced in 76 patients. Equine serum, although giving rise to more reactions, was the only effective agent in terms of the levels and duration of serum antitoxin concentration produced. The local response to a test dose of any of the three sera is not a reliable guide to immediate or late general reactions.

A survey of the species composition and distribution of the Culicoides midge populations at a range of sites where bluetongue is enzootic isolated a group of dominant species: C. cornutus, C. grahamii, C. magnus, C. milnei, C. pallidipennis and C. 23.† Monthly light-trap sampling of Culicoides showed that the population densities of the dominant species greatly increased after the rain seasons and that these species concentrated around flocks of sheep and cattle. The larval habitats of C. cornutus and C. pallidipennis were found associated with stock pens. Precipitin tests on blood-fed Culicoides showed that most of the dominant species regularly feed on sheep and cattle. Bluetongue virus was isolated from C. milnei, C. pallidipennis and C. 23. Serological surveys of wild and domestic bovids from the enzootic area showed a high proportion with antibody to bluetongue virus. The colonization of C. cornutus, a potential vector, is described briefly. A causal relationship between peak rainfall in April-May, peak numbers of Culicoides in May-June and peak bluetongue incidence in June-July is postulated. The vector status of the above species and C. austeni was evaluated.

A streptococcal antigen that is closely associated with the M-antigen, but is not type specific can be detected by means of a complement-fixation test in extracts of M-positive, but not of M-negative, variants of group A streptococci. Purification of acid extracts results in a concomitant increase in the purity both of the type-specific M-antigen and of the M-associated protein (MAP). Antibody to MAP is present in the sera of patients who have had streptococcal infection. The highest titres are found in patients with rheumatic fever.

When fresh animal serum was dropped onto seeded mycoplasma agar plates, inhibition of growth frequently occurred. This effect was dependent on the mycoplasma serotype and on the animal species from which the fresh serum came. This activity of fresh animal serum was heat-labile and would not diffuse through the agar medium. Growth of all the porcine mycoplasma serotypes was inhibited by fresh sheep serum. M. hyorhinis, M. hyopneumoniae, B 3 and the P 45 strains were insensitive to fresh horse serum. The addition of fresh horse serum to specific M. hyorhinis rabbit antiserum-impregnated disks appeared to have a synergistic effect and the combination of M. hyorhinis antiserum-impregnated disk and fresh horse serum always inhibited the growth of M. hyorhinis strains.