Co-cultivation of Legionella pneumophila serogroup I and Acanthamoeba palestinensis in Neff's medium at 35 °C resulted in the intracellular multiplication of the bacteria as demonstrated by electron microscopy and immunofluorescence. In the closed experimental system used, the number of legionellae rose from 107 colony forming units (c. f. u.)/ml initially to a maximum of 1010 c. f. u. /ml on day 5. Legionellae were seen in expelled phagosomes, in some amoebae filling the cytoplasm and in others in which the process of encystment appeared to have commenced.

A systematic review of studies providing frequency estimates of brucellosis in humans and ruminants and risk factors for Brucella spp. seropositivity in humans in the Middle East was conducted to collate current knowledge of brucellosis in this region. Eight databases were searched for peer-reviewed original Arabic, English, French and Persian journal articles; the search was conducted on June 2014. Two reviewers evaluated articles for inclusion based on pre-defined criteria. Of 451 research articles, only 87 articles passed the screening process and provided bacteriological and serological evidence for brucellosis in all Middle Eastern countries. Brucella melitensis and B. abortus have been identified in most countries in the Middle East, supporting the notion of widespread presence of Brucella spp. especially B. melitensis across the region. Of the 87 articles, 49 were used to provide evidence of the presence of Brucella spp. but only 11 provided new knowledge on the frequency of brucellosis in humans and ruminants or on human risk factors for seropositivity and were deemed of sufficient quality. Small ruminant populations in the region show seroprevalence values that are among the highest worldwide. Human cases are likely to arise from subpopulations occupationally exposed to ruminants or from the consumption of unpasteurized dairy products. The Middle East is in need of well-designed observational studies that could generate reliable frequency estimates needed to assess the burden of disease and to inform disease control policies.

At the present time, COVID-19 is spreading rapidly [1]. The global prevention and control of COVID-19 is focused on the estimation of the relevant incubation period, basic reproduction number (R0), effective reproduction number (Rt) and death risk. Although the prevention and control of COVID-19 requires a reliable estimation of the relevant incubation period, R0, Rt and death risk. Another key epidemiological parameter-asymptomatic ratio that provides strength and range for social alienation strategies of COVID-19, which is widely defined as the proportion of asymptomatic infections among all disease infections. In fact, the ratio of asymptomatic infection is a useful indicator of the burden of disease and a better measurement of the transmissibility of the virus. So far, people have not paid enough attention to asymptomatic carriers. The asymptomatic carriers discussed in this study are recessive infections, that is, those who have never shown symptoms after onset of infection. We will discuss three aspects: detection, infectivity and proportion of healthy carriers.

This paper analyses Streptococcus pneumoniae transmission dynamics in households using longitudinal data on pneumococcal (Pnc) carriage in the United Kingdom. Ten consecutive swabs were taken at 4-week intervals from all members of 121 households. The family status is derived from the observed Pnc carriage status of each family member. Transition matrices are built for each family size and composition containing the observed frequency of transitions between family statuses over a 28-day interval. A density-dependent transmission model is fitted to derive maximum-likelihood estimates of the duration of carriage and acquisition rates from the community and from infected individuals within the household. Parameter values are estimated for children (<5 years) and adults (5+ years). The duration of carriage is longer in children <5 years of age than in older family members (51 vs. 19 days). Children are 3–4 times more likely than adults to acquire Pnc infection from the community. Transmission rates within the household suggest that adults are more infectious but less susceptible than children. Transmission within the household is most important in large families. The proportion of household-acquired infection ranges from 29 to 46% in households of three persons to 38–50% in larger households. Evidence of density-dependent within-household transmission is found, although the strength of this relationship is not clear from the model estimates.

Between 1 January 2009 and 31 December 2012 in England, a total of 3717 cases were reported with evidence of Shiga toxin-producing E. coli (STEC) infection, and the crude incidence of STEC infection was 1·80/100 000 person-years. Incidence was highest in children aged 1–4 years (7·63/100 000 person-years). Females had a higher incidence of STEC than males [rate ratio (RR) 1·24, P < 0·001], and white ethnic groups had a higher incidence than non-white ethnic groups (RR 1·43, P < 0·001). Progression to haemolytic uraemic syndrome (HUS) was more frequent in females and children. Non-O157 STEC strains were associated with higher hospitalization and HUS rates than O157 STEC strains. In STEC O157 cases, phage type (PT) 21/28, predominantly indigenously acquired, was also associated with more severe disease than other PTs, as were strains encoding stx2 genes. Incidence of STEC was over four times higher in people residing in rural areas than urban areas (RR 4·39, P < 0·001). Exposure to livestock and/or their faeces was reported twice as often in cases living in rural areas than urban areas (P < 0·001). Environmental/animal contact remains an important risk factor for STEC transmission and is a significant driver in the burden of sporadic STEC infection. The most commonly detected STEC serogroup in England was O157. However, a bias in testing methods results in an unquantifiable under-ascertainment of non-O157 STEC infections. Implementation of PCR-based diagnostic methods designed to detect all STEC, to address this diagnostic deficit, is therefore important.

A modification of the Henderson apparatus is described which is mobile and which can produce bacterial or viral aerosol in air whose relative humidity can be chosen in the range 20–95 %. The apparatus forms a basic utility for microthread, rotating drum or other aerosol studies.

Much help and advice has been given by Messrs. J. F. H. Peel and D. J. Garriock of the Engineering Section M.R.E. throughout the development of this apparatus. The proportioning valve is a modification of one originally used for a similar purpose by Mr K. R. May. Thanks are due to Messrs. G. J. Harper and A. M. Hood for criticism of the script.

Strains of myxoma virus associated with a case-mortality rate in wild rabbits of 90—95% can be distinguished from those associated with a case-mortality rate of more than 99% by the use of a standardized virulence test carried out in groups of five laboratory rabbits, and tests carried out on laboratory rabbits are closely correlated with the results obtained with Australian wild rabbits. The test, which attempts to mimic natural mosquito transmission, consists of the intradermal inoculation in one site of about 5 rabbit-infectious doses of the virus, and the subsequent observation of the rate of progress of clinical symptoms and ultimate fate of the rabbit. The mean survival time provides a figure which allows satisfactory classification of all known myxoma virus strains into one of four grades of virulence, very attenuated strains being recognized by their low case-mortality rates.

Using this test, 92 strains of myxoma virus have been compared. These comprised 6 laboratory strains, 62 strains recovered from Australian wild rabbits or from wild caught mosquitoes, between February, 1951 and March, 1955, 19 strains recovered from naturally infected rabbits in Europe between October, 1953 and August, 1955, 3 strains from South America, and 2 from California. Eight types have been described and illustrated, differentiation being based on their virulence and the clinical picture of the disease they cause in laboratory rabbits. The differences are summarized in Text-fig. 2. Four of the virus types produce recognizably different pocks on the chorioallantois of developing chick embryos.

In Australia the repeatedly introduced standard laboratory strain of virus has been replaced by a moderately virulent strain, which appears to be better adapted for survival than either the highly virulent standard laboratory strain, or less virulent variants of which only a few had been recovered up to 1955. Strains of the type dominant in 1953–55 have appeared independently in many widely separated parts of Australia. Examination of these attenuated variants by pure clone techniques showed that the variability in symptomatology associated with them is not due to a mixture of virus strains, but to the expression of differences in innate resistance of the host animals which are obscured by the overwhelming virulence of the highly virulent strains.

Only one introduction of a different strain of virus was made in Europe. All field strains recovered during the first two years after the introduction were of high virulence, but a few attenuated strains appeared in 1955, and they have since become widespread. One of them appears to be a relatively stable mixture of a highly virulent and a greatly attenuated strain.

The collection of the large numbers of strains of virus described in this paper has been made possible only by the generous collaboration of many people. In Australia we would like to acknowledge the great help afforded us by Mr F. N. Ratcliffe, officer-in-charge of the Wildlife Survey Section, Commonwealth Scientific and Industrial Research Organization and his officers, especially Messrs J. H. Calaby, G. Douglas, A. L. Dyce, B. V. Fennessy, E. W. Lines, K. Myers, R. Mykytowycz, W. E. Poole and E. Waterhouse. Messrs B. D. Robinson and T. Pearce of the Lands Department of Victoria have also provided us with specimens, as have Mr G. Edgar, Director of the Veterinary Research Station, Glenfield, New South Wales, Dr D. Surrey Dane of the Institute of Medical and Veterinary Science, Adelaide, Mr D. Gooding, Department of Agriculture, Western Australia, and Messrs T. B. Alexander and A. F. Ryan, Department of Agriculture, Tasmania.

We are indebted to Dr H. de B. Aragão of the Instituto Oswaldo Cruz, Rio de Janeiro, and Dr B. Szyfres, of the Laboratorio de Biologia Animal, Uruguay, for specimens from South America; Dr D. G. McKercher of the University of California School of Veterinary Medicine, Davis, California, for the Californian strains; Drs P. Lépine and H. Jacotot, of the Institut Pasteur, Paris, for strains from France; and Mr J. R. Hudson, of the Veterinary Laboratory, Weybridge, and Dr C. H. Andrewes of the National Institute for Medical Research, Mill Hill, for material from England.

We are grateful to Dr S. Fazekas de St Groth and Dr G. S. Watson of this University for advice on the statistical treatment of the data.

Mr V. Paral took the photographs.

Faeces samples from diarrhoeic and non-diarrhoeic lambs and goat kids aged 1–45 days were examined for enteric pathogens. Cryptosporidium parvum was detected in both diarrhoeic lambs (45%) and goat kids (42%) but not in non-diarrhoeic animals. F5+ (K99+) and/or F41+Escherichia coli strains were isolated from 26% and 22% of the diarrhoeic lambs and goat kids, respectively, although these strains, which did not produce enterotoxins ST I or LT I, were found with similar frequencies in non-diarrhoeic animals. A F5−F41−ST I+E. coli strain was isolated from a diarrhoeic lamb (0·6%). Verotoxigenic E. coli was isolated from both diarrhoeic and non-diarrhoeic lambs (4·1% and 8·2%, respectively) and there was no association between infection and diarrhoea. The prevalence of group A rotavirus infection in diarrhoeic lambs was very low (2·1%). Groups A and B rotaviruses were detected in three (8·1%) and five (13·5%) diarrhoeic goat kids from two single outbreaks. Group C rotaviruses were detected in four non-diarrhoeic goat kids. An association of diarrhoea and infection was demonstrated only for group B rotavirus. Clostridium perfringens was isolated from 10·8% of the diarrhoeic goat kids but not from non-diarrhoeic goat kids or lambs. Salmonella arizonae was isolated from a diarrhoeic goat kid (2·7%) and the clinical characteristics of the outbreaks where these two latter enteropathogens were found different from the rest. Picobirnaviruses were detected in a diarrhoeic lamb. No coronaviruses were detected using a bovine coronavirus ELISA. No evidence was found of synergistic effect between the agents studied. Enteric pathogens were not found in four (8·7%) and three (20%) outbreaks of diarrhoea in lambs and goat kids, respectively.

This systematic review aimed at estimating chronic hepatitis B (HBV) and C virus (HCV) prevalence in the European Union (EU) and Economic Area (EEA) countries in the general population, blood donors and pregnant women. We searched PubMed©, Embase© and Cochrane Library databases for reports on HBV and HCV prevalence in the general population and pregnant women in EU/EEA countries published between 2005 and 2015. Council of Europe data were used for HBV and HCV blood donor prevalence. HBV general population estimates were available for 13 countries, ranging from 0·1% to 4·4%. HCV general population estimates were available for 13 countries, ranging from 0·1% to 5·9%. Based on general population and blood donor estimates, the overall HBV prevalence in the EU/EEA is estimated to be 0·9% (95% CI 0·7–1·2), corresponding to almost 4·7 million HBsAg-positive cases; and the overall HCV prevalence to be 1·1% (95% CI 0·9–1·4), equalling 5·6 million anti-HCV-positive cases. We found wide variation in HCV and HBV prevalence across EU/EEA countries for which estimates were available, as well as variability between groups often considered a proxy for the general population. Prevalence estimates are essential to inform policymaking and public health practice. Comparing to other regions globally, HBV and HCV prevalence in the EU/EEA is low.

A model for contact transfer of micro-organisms by hand has been extended to include representatives of bacterial species responsible for a majority of hospitalacquired infections. The ability of the organisms to transfer from contaminated fabrics to hands and from hands to sterile fabrics was measured, as was their ability to survive on the skin of the hands. There were differences between the species. Staphylococcus saprophyticus transferred well to the hand but not as well from hand to fabric as the other species; it survived well on skin. Pseudomonas aeruginosa, Klebsiella aerogenes and Serratia marcescens transferred moderately well overall and also survived on the skin. These results were in contrast to those obtained with a strain of Escherichia coli and one of Streptococcus pyogenes.

The contact transfer model was used to investigate the use of small volumes of alcohol in preventing transfer via the hands. An alcohol handrub of either 0·3 ml 80% ethanol or 0·3 ml 70% isopropanol gave reductions in transfer slightly less than that of a soap and water wash. Raising the volume, and consequently the contact time, to 0·5 ml 70% isopropanol gave a 14000-fold reduction in transfer, statistically indistinguishable from that of a thorough soap and water wash (9800-fold reduction).

The zoonotic risk of Brucella canis has been considered fairly high for persons who handle breeding dogs in kennels or are exposed to infected animals. Transmission to humans in other circumstances has been thought to be rare. We describe an uncommon outbreak of brucellosis caused by B. canis which, to the best of our knowledge, is the first reported in the literature. This outbreak involved six persons (three children and three adults), a bitch and three puppies which had close daily contact with the family. The clinical symptoms of the index case led to an erroneous diagnosis and the infection would have gone undiagnosed if culture had not been positive. This report aims to increase awareness of medical personnel of the need to order screening tests for children, immunodeficient persons or pregnant women presenting with fever of unknown origin, unexplained spleen or liver enlargement or other systemic signs. The emerging zoonotic potential of this disease in urban areas and the need to coordinate canine brucellosis surveillance systems should be evaluated.

To test the hypothesis that chronic infection with Chlamydia pneumoniae, Helicobacter pylori or cytomegalovirus is associated with coronary heart disease risk in elderly men, a nested case-control study in a cohort investigated in 1985 and 1990 in the town of Zutphen, The Netherlands, was designed. Fifty-four cases with a first diagnosed coronary event between 1985 and 1990, and 108 age-matched control subjects free of coronary heart disease during follow up were included in the study. The overall prevalence of antibodies to cytomegalovirus was 74·7%, to H. pylori 75·9% and to C. pneumoniae 84·0%. A high level of antibodies to C. pneumoniae was associated with an increased coronary heart disease risk (OR=2·76; 95% CI=1·31–5·81). This association was stronger in cases developing both myocardial infarction and angina pectoris, than in cases developing only one of these. This association was independent of potential confounders. Antibodies to cytomegalovirus or H. pylori were not associated with coronary heart disease risk. These results support the hypothesis of a role of chronic C. pneumoniae infections in the immunopathogenesis of atherosclerosis.

Nose, throat and finger carriage of Staphylococcus aureus was investigated in a series of random samples from a normal European population.

No evidence for a seasonal trend in carriage was found but the intersample variation between successive random samples was obtained. The mean nasal carrier rate was 29 % with a standard deviation of 7 %.

No association was found between nasal or throat carriage of staphylococci and stay in hospital or antibiotic therapy but respondents with penicillin-resistant staphylococci in the nose had skin infections more frequently than those with penicillin-sensitive strains.

Evidence was obtained for a family, perhaps genetic, ‘predisposition’ to carry staphylococci in the nose.

We investigated age- and gender-specific incidence of shingles reported in a large sentinel practice network monitoring a defined population over the years 1994–2001. In total, 5915 male and 8617 female incident cases were studied. For each age group, we calculated the relative risk of females to males presenting with shingles. Incidence rates of chickenpox and herpes simplex were examined similarly. Shingles incidence was greater in females in each age group (except for 15–24 years). Relative risks (female to male) were greatest in age groups 45–64 years (1·48) and 0–14 years (1·43). There were no gender differences in the incidence of chickenpox except in the 15–24 years age group (female excess): for herpes simplex there were female excesses in all age groups. Gender-specific age-standardized incidence rates of shingles were calculated for each year and showed a consistent female excess in each of the 8 years (average annual excess 28%).

Our study aimed to systematically analyse the risk factors of coronavirus disease 2019 (COVID-19) patients with severe disease. An electronic search in eight databases to identify studies describing severe or critically ill COVID-19 patients from 1 January 2020 to 3 April 2020. In the end, we meta-analysed 40 studies involving 5872 COVID-19 patients. The average age was higher in severe COVID-19 patients (weighted mean difference; WMD = 10.69, 95%CI 7.83–13.54). Patients with severe disease showed significantly lower platelet count (WMD = −18.63, 95%CI −30.86 to −6.40) and lymphocyte count (WMD = −0.35, 95%CI −0.41 to −0.30) but higher C-reactive protein (CRP; WMD = 42.7, 95%CI 31.12–54.28), lactate dehydrogenase (LDH; WMD = 137.4, 95%CI 105.5–169.3), white blood cell count（WBC), procalcitonin（PCT）, D-dimer, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and creatinine（Cr）. Similarly, patients who died showed significantly higher WBC, D-dimer, ALT, AST and Cr but similar platelet count and LDH as patients who survived. These results indicate that older age, low platelet count, lymphopenia, elevated levels of LDH, ALT, AST, PCT, Cr and D-dimer are associated with severity of COVID-19 and thus could be used as early identification or even prediction of disease progression.

Recent much-publicized increases in pertussis case reports in some countries with high vaccine coverage have raised concerns about its current and future control. The ubiquity of this trend, however, remains unexamined. In an attempt to paint a global picture, we used case counts to determine which countries experienced statistically significant trends in incidence over the past two decades and to map changes in incidence during this period. These data reveal that pertussis resurgence is not a universal phenomenon. The heterogeneity in incidence trends, even in countries with superficially similar demography, socioeconomic conditions and vaccination programmes, is striking and requires explanation. In this opinion piece, we review and assess the multifaceted proposed explanations incorporating evolution, population dynamics, and the details of immunization programmes. While we do not solve the riddle that is pertussis epidemiology, we highlight critical aspects that are likely to hold the key to understanding its worldwide epidemiology.

1. Previous work on the antigenic structure of the cholera group of vibrios is experimentally reviewed, and the data amplified and systematised.

2. The cholera group is defined as consisting of vibrios with similar biochemical characters and having a common heat-labile antigenic component.

3. The heat-stable antigens are divisible into:

(a) A considerable number of specific antigens, best demonstrated by O sera and H-O suspensions, which serve as a basis of classification into O subgroups.

(b) A non-specific component, demonstrable with O sera and O suspensions.

4. The first subgroup contains all the standard cholera vibrios from central laboratories, and the majority of other epidemic strains. We consider that it represents the only class of vibrios known for certain to cause epidemic cholera.

5. The races of this subgroup I are further divisible into two (or perhaps three) “types”, as established by Japanese workers, according to differences in their subsidiary O antigens.

6. The haemolytic “El Tor” vibrios are serologically diverse. The term “El Tor” should, as Shousha suggests, be reserved for those that have the same specific O component as the standard cholera vibrios.

7. For the identification of the undoubted cholera vibrios a standard subgroup I O serum is recommended in conjunction with the haemolytic test. The serum should contain both the main and the subsidiary antigens of the subgroup.

8. As a working rule it is suggested that bacteriological proof of “cholera” or a cholera carrier should rest on the isolation of a non-haemolytic vibrio with the specific O antigen of subgroup I.

The studies and observations on which this paper is based were conducted while one of us (K. V. V.) was holding a Fellowship of the International Health Division of the Rockefeller Foundation.

Our thanks are due to Mr P. Bruce White for constant help and advice, and to the numerous bacteriologists in various lands who have kindly supplied us with cultures and sera.

In this study, we evaluated the association between high-risk human papillomavirus (hrHPV) and the vaginal microbiome. Participants were recruited in Nigeria between April and August 2012. Vaginal bacterial composition was characterized by deep sequencing of barcoded 16S rRNA gene fragments (V4) on Illumina MiSeq and HPV was identified using the Roche Linear Array® HPV genotyping test. We used exact logistic regression models to evaluate the association between community state types (CSTs) of vaginal microbiota and hrHPV infection, weighted UniFrac distances to compare the vaginal microbiota of individuals with prevalent hrHPV to those without prevalent hrHPV infection, and the Linear Discriminant Analysis effect size (LEfSe) algorithm to characterize bacteria associated with prevalent hrHPV infection. We observed four CSTs: CST IV-B with a low relative abundance of Lactobacillus spp. in 50% of participants; CST III (dominated by L. iners) in 39·2%; CST I (dominated by L. crispatus) in 7·9%; and CST VI (dominated by proteobacteria) in 2·9% of participants. LEfSe analysis suggested an association between prevalent hrHPV infection and a decreased abundance of Lactobacillus sp. with increased abundance of anaerobes particularly of the genera Prevotella and Leptotrichia in HIV-negative women (P < 0·05). These results are hypothesis generating and further studies are required.

The reservoir host of cowpox virus in Western Europe is not known, but epidemiological evidence from human and feline infections indicates that the virus is probably endemic in small wild rodents. Therefore, serum and tissue samples were collected from a variety of wild British mammals and some birds, and tested for evidence of Orthopoxvirus infection. Antibody reacting with cowpox virus was detected in 9/44 (20%) bank voles (Clethrionomys glareolus), 8/24 (33%) field voles (Microtus agrestis), 17/86 (20%) wood mice (Apodemus sylvaticus) and 1/44 house mice (Mus musculus), but in no other animal species tested. Although virus was not isolated from any animal, this serological survey, together with other evidence, suggests that bank and field voles and wood mice are the main reservoir hosts of cowpox virus in Great Britain.

The effects of temperature on reported cases of a number of foodborne illnesses in England and Wales were investigated. We also explored whether the impact of temperature had changed over time. Food poisoning, campylobacteriosis, salmonellosis, Salmonella Typhimurium infections and Salmonella Enteritidis infections were positively associated (P<0·01) with temperature in the current and previous week. Only food poisoning, salmonellosis and S. Typhimurium infections were associated with temperature 2–5 weeks previously (P<0·01). There were significant reductions also in the impact of temperature on foodborne illnesses over time. This applies to temperature in the current and previous week for all illness types (P<0·01) except S. Enteritidis infection (P=0·079). Temperature 2–5 weeks previously diminished in importance for food poisoning and S. Typhimurium infection (P<0·001). The results are consistent with reduced pathogen concentrations in food and improved food hygiene over time. These adaptations to temperature imply that current estimates of how climate change may alter foodborne illness burden are overly pessimistic.