Resting Position of the Imago.—Geographical Distribution of the Species.—Habitat.—Modes of Dissemination and Migratory Flights.—hibernation of the Imago.—Longevity.—Oviposition, and the Influence of Food thereon.—Parthenogenesis.—Numerical Proportion of Sexes.—Number of Generations during a season.—Food.—Influence of Heat, Cold, Light, Colour, and Sound.—Sense of Smell and Taste.

The concomitant occurrence of a case of haemolytic–uraemic syndrome (HUS) and 62 cases of mild gastroenteritis in schools of a small rural community in southern Italy induced the health authorities to suspect a foodborne outbreak of shiga-toxin-producing Escherichia coli (STEC) infection. The schools were closed and the catering service involved was investigated. However, STEC were not isolated from the HUS case or from the 56 cases of gastroenteritis examined, and the HUS case and the outbreak of gastroenteritis were probably just coincidental. A retrospective cohort study failed to show any correlation with consumption of school meals and suggested that the outbreak probably started outside the school setting and then spread within the schools by person-to-person transmission. All the cases examined were negative for common enteric pathogens and the responsible agent for the cases of gastroenteritis was not identified. The concern raised in the small community by the occurrence of a severe case of HUS and the lack of a rapid epidemiological assessment excluding the occurrence of a STEC outbreak, turned an epidemic episode of mild gastroenteritis into a public health emergency with relevant socioeconomic consequences. Prompt intervention in outbreaks following timely and effective risk communication are crucial for taking the most appropriate control measures and avoiding the spread of fear and panic in the community.

1. An agar medium is described which enables a direct estimation of the numbers of coliform bacilli in a water or milk sample to be made.

2. An enrichment medium for B. lactis aerogenes is one containing in every 100 c.c. of peptone water 0·5 g. sodium citrate, 0·25 g. soluble starch and 1 c.c. of a 1 per cent, watery solution of neutral red.

3. Enrichment media for B. coli are: (a) lactose peptone water containing in every 100 c.c., 1 g. sodium sulphite anhydrous and 2 c.c. of 1 per cent, rosolic acid in absolute alcohol; (b) lactose peptone water containing in every 100 c.c., 2 c.c. of a 40 per cent, solution of hexamine in water.

4. Importance is attached to the capacity to ferment starch possessed by B. lactis aerogenes.

5. The application of these media to the examination of water and milk is indicated.

The aim of this study was to assess the effects of reductions in the number of isolates tested by phage-typing on the recognition of outbreaks of salmonellosis. Five outbreaks (categorized as ‘small’, ‘medium’ or ‘large’) which occurred in England in 2005 were used as examples. The outbreaks were caused by serotypes which were subdivided by phage-typing. Results indicated that reducing the number of isolates phage-typed would have an impact on the surveillance system, with one outbreak likely to have been missed altogether. However, this does not have a great effect on the ‘time-to-detection’ for the other outbreaks. Assuming no testing for phage-typing was undertaken it is likely that two out of five outbreaks would not have been detected. Assessing the value of phage-type information is important not only in deciding on the efficiency of the current surveillance system but also in providing a basis upon which to assess more detailed typing methodologies such as an antibiogram of molecular profile.

In England, there is no surveillance system for vaccines offered to healthcare workers apart from that in place annually for the seasonal influenza vaccine. To inform the feasibility of a general vaccine uptake surveillance system and to understand which policies are currently in place, we conducted a survey in the 162 National Health Service Foundation and Acute Hospital trusts in England, by submitting a questionnaire to their occupational health departments on immunization policies and methods of storing vaccine uptake data. In total, 104 hospital trusts (64·2%) responded. All responders offer hepatitis B, tuberculosis, measles-mumps-rubella, and influenza vaccines to healthcare workers; 0·9% reported not offering varicella and 13·5% not offering tetanus-diphtheria-polio; 66·4% record staff eligible for immunizations and 68·2% record staff they have immunized. Our study suggests that setting up a surveillance system to monitor vaccine uptake in healthcare workers is possible but would be challenging, given the variation in current systems.

An area of the laboratory bench on which slide agglutinations were performed in the diagnosis of salmonella and shigella infection was examined for these organisms. Impression plates and broth-moistened swabs were used for sampling. Both techniques gave satisfactory results, but the contact plates provided positive results a day earlier than the swabs. Suitable precautions to minimize contamination of the bench surface are discussed.

Salmonella enterica commonly colonizes the intestinal tract of cattle and is a leading cause of foodborne illness. A previously described investigation into the prevalence of S. enterica on a dairy farm revealed an 8-year-long asymptomatic S. enterica epidemic caused by serotypes Cerro and Kentucky in the lactating herd. To investigate the source of the S. Kentucky strains, the genomes of two S. Kentucky isolates were sequenced; one collected prior to the epidemic (2004) and one collected during the epidemic (2010). Comparative genomic analysis demonstrated significant polymorphisms between the two strains. PCR primers targeting unique and strain-specific regions were developed, and screening of the archived isolates identified the index case of the asymptomatic S. Kentucky epidemic as a heifer that was raised off-site and transported onto the study farm in 2005. Analysis of isolates collected from all heifers brought onto the farm demonstrated frequent re-introduction of clones of the epidemic strain suggesting transmission of pathogens between farms might occur repeatedly.

This study aimed to determine the prevalence of serological markers for HIV-1/2, HBV, HCV, Treponema cruzi and T. pallidum infections. The association of these infections with risk factors in a population from Salvador, Bahia, Brazil was also analysed. Of the 780 enrolled individuals, 545 (70%) were female and 235 (30%) were male. Seroprevalence of 0·8% (6/702), 1·3% (9/678), 1·5% (10/684), 3·5% (23/663) and 11·5% (77/668) for HIV-1/2, HBV, HCV, T. cruzi and T. pallidum infections, respectively, was observed. The seroprevalence of T. pallidum was higher in males 20% (43/210) than in females 7% (34/458) (P < 0·01). An association between age and seroprevalence for T. cruzi (P = 0·02) and T. pallidum (P < 0·01) was observed. HBsAg was associated with having tattoos (3/37 vs. 6/623, P = 0·01) and not having a steady sexual partner (5/141 vs. 4/473, P = 0·04), while anti-HIV-1/2 was associated with having tattoos (2/39 vs. 4/647, P = 0·04); however, larger studies are needed to categorically state the relationship of these risk factors with infectious agents. The prevalence of serological markers for HIV-1/2, HBV, HCV and T. cruzi was consistent with other studies.

According to Kossel (1888) Fritz Miiller discovered that phthisical sputum digests fibrin and coagulated albumen in weakly alkaline media, but the fact that antiseptic pus digests proteins seems to have been first observed by Leber.

Achalme (1899) separated proteolytic ferments from pus from various sources and Erben (1903) and Schumm (1904) drew attention to the notable quantity of albumoses in blood from cases of myologenous leukaemia which had been incubated, and the relative absence of this proteolysis in the incubated blood from cases of lymphatic leukaemia. Ascoli and Mareschi (1901) showed that the peritoneal exudate produced by aleuronat underwent autolysis under aseptic conditions.

1. When coagulase-positive staphylococci are grown on nutrient agar containing fresh mammalian serum and haemoglobin, discrete areas of clearing appear in the medium near to the growth.

2. These ‘plaques’ are due to proteolysis, the haemoglobin acting as a chromoprotein indicator substrate. For convenience the effect is referred to as ‘proteoclast’.

3. The effect is due to the interaction of a diffusible product of the staphylococci with constituents of normal sera. The staphylococcal factor has been obtained in a bacterium-free filtrate and its interaction with serum studied quantitatively. It resists boiling.

4. The number of plaques produced under defined conditions showed a linear correlation with the amount of serum used, but not with the amount of filtrate.

5. By means of serial filtration through Gradocol membranes it was shown that one factor in normal serum required for the proteoclast effect is particulate, having a size of the order 20–70 m μ diameter.

6. It was found that a third factor present in fresh sera was also required.

7. Serum was rendered inactive by conditions of heating and ultra-violet irradiation similar to those required to sterilize vegetative bacteria and viruses.

8. It is suggested that the proteoclast effect is due either to a non-pathogenic virus, or to a hitherto unrecognized particulate enzyme precursor present in sera and activated by the staphylococcal factor.

Outbreaks caused by Chlamydia psittaci and other chlamydial species have recently been reported in poultry farms worldwide, causing considerable economic losses. The objective of this study was to determine the presence of chlamydial species in these birds in Costa Rica. One hundred and fifty pools of lung tissue samples from industrial poultry with respiratory problems and 112 pools of tracheal swabs from asymptomatic backyard poultry were analysed by real-time quantitative polymerase chain reaction (qPCR), end-point PCR and sequencing. A total of 16.8% (44/262) samples were positive for Chlamydia spp., most of them detected in asymptomatic backyard poultry (28.6%, 32/112) and fewer in industrial poultry (8%, 12/150). Of these positive samples, 45.5% (20/44) were determined to be C. psittaci. For the first time C. psittaci genotype A is reported in poultry in Latin America. In addition, the presence of Chlamydia gallinacea in backyard poultry and of Chlamydia muridarum in industrial and backyard poultry is reported for the first time in Central America. In 40.9% (18/44) of the positive samples, it was not possible to identify the infecting chlamydial species. These findings reveal a zoonotic risk, particularly for poultry farm and slaughterhouse workers having direct contact with these birds.

(1) Our experiments support the view that influenza is casued by B. influenzae, and that pneumococci and certain streptococci are the most important agents of secondary infections. No new type of bacterium was discovered, and no evidence was obtained of the presence of an ultra-micrascopic virus.

(2) B. influenzae appears more toxic for rabbits and mice when grown on fresh liver or kidney media than when grown on blood agar or in blood broth, and this is especially so when the kidney or liver is obtained from an animal of t he same species that has previously been inoculated with a culture of the influenza bacillus.

(3) Three fairly distinct special forms have been isolated from cultures of B. dysenteriae. They are probably not sexual units or stages in a true life cycle. Special large forms have also been obtained from cultures of B. influenzae, but these reverted back to the normal small type after several sub-cultures.

(4) A filter-passing material has been found associated with certain micro- cocci from vaccinia and in pure cultures of members of the dysentery-typhoid- coli group of bacteria. This material breaks down and dissolves the bacteria of the cultures, and the “infection” can be carried to fresh normal cultures. The evidence is against its being a living ultra-microscopic virus infecting the bacteria, but it may have some connection with the special forms. No definite evidence of a similar dissolving material has been found associated with cultures of the influenza bacillus.

(5) There is some indication that one or more of the forms isolated from pure cultures of dysentery bacilli may be special toxin-producing units, also that this function is at its maximum when the bacilli are first produced by the normal small forms or by sexual units of a normal culture. This may possibly be the case also with special forms that have been found in cultures of the influenza bacillus.

(6) It is believed that non-pathogenic wild varieties of ultra-microscopic viruses must exist in nature, and that they should present less difficulty in cultivation than the pathogenic varieties. Cultivations were made from filtrates of soil, faeces and water on various special media, and the tubes incubated in various gases. The chief gases tested either alone or mixed were oxygen, nitrogen, carbon dioxide and sulphuretted hydrogen. All the results were negative, but some interesting deposit colonies were obtained on media con taining a small quantity of sodium sulphide.

(7) The influenza bacillus grows in symbiosis with amoebae on blood agar, and in such cultures the bacillus lives considerably longer.

(8) The influenza bacillus grows in symbiosis with a small spirillum that was isolated from a grass emulsion, and in symbiosis with an extremely minute and delicate bacterium isolated from garden soil; after passing the emulsions through a Berkefeld filter. With these delicate bacteria the influenza bacillus will grow on media containing no blood, and if an egg medium is used the mixed growth is dense, heaped up and brownish in colour. In such cultures the in fluenza bacillus may live for months.

(9) The influenza and other bacilli will grow in an atmosphere of pure oxygen or pure carbon dioxide under diminished pressure.

The history of the Brown Animal Sanatory Institution was intended originally to be published in book form, and was therefore divided into chapters. For the sake of convenience this division has been continued, even though it is unusual for the term to be employed in scientific periodicals. The history will be published serially in four consecutive issues of the Journal of Hygiene. References have presented a difficulty. In two instances the title of the paper referred to has had to be omitted, because no copy of the original journal in which the article appeared has been obtainable. More important, however, is the failure to give references, beyond the date, to a number of statements in the text, because the information in question has been abstracted from sources that are not available to the general reader.

A total of 111 clinical and environmental O1, O139 and non-O1/O139 Vibrio cholerae strains isolated between 1978 and 2008 from different geographical areas were typed using a combination of methods: antibiotic susceptibility, biochemical test, serogroup, serotype, biotype, sequences containing variable numbers of tandem repeats (VNTRs) and virulence genes ctxA and tcpA amplification. As a result of the performed typing work, the strains were organized into four clusters: cluster A1 included clinical O1 Ogawa and O139 serogroup strains (ctxA+ and tcpA+); cluster A2 included clinical non-O1/O139 strains (ctxA− and tcpA−), as well as environmental O1 Inaba and non-O1/O139 strains (ctxA− and tcpA−/tcpA+); cluster B1 contained two clinical O1 strains and environmental non-O1/O139 strains (ctxA− and tcpA+/tcpA−); cluster B2 contained clinical O1 Inaba and Ogawa strains (ctxA+ and tcpA+). The results of this work illustrate the advantage of combining several typing methods to discriminate between clinical and environmental V. cholerae strains.

Beijing–Tianjin–Hebei is the largest urban agglomeration in northern China, but the spatiotemporal patterns and risk factors concerning hepatitis B virus (HBV) incidence in this area have been unclear. The present study aimed to reveal the spatiotemporal epidemiological features of HBV infection and quantify the association between HBV infection and socio-economic risk factors. The data on HBV cases in Beijing–Tianjin–Hebei from 2007 to 2012 was collected for each county. The Bayesian space–time hierarchy model and the GeoDetector method were used to reveal spatiotemporal patterns and detect risk factors. High-risk regions were mainly distributed in the underdeveloped rural areas in the north and mid-south of the study region, while low-risk regions were mainly distributed in the urban and western areas. The HBV annual incidence rate decreased substantially over the 6-year period, dropping from 7.34/105 to 5.51/105. Compared with this overall trend, 38.5% of high-risk counties showed a faster decrease, and 35.9% of high-risk counties exhibited a slower decrease. Meanwhile, 29.7% of low-risk counties had a faster decrease, and 44.6% of low-risk counties exhibited a slower decrease. Socio-economic factors were strongly associated with the spatiotemporal patterns and variation. The population density and gross domestic product per capita were negatively associated with HBV transmission, with determinant powers of 0.17 and 0.12, respectively. The proportion of primary industry and the number of healthcare workers were positively associated with the disease incidence, with determinant powers of 0.11 and 0.8, respectively. The interactive effect between population density and the other factors exerted a greater influence on HBV transmission than that of these factors measured independently.

Rabbit polyclonal hyperimmune antibodies to Yersinia pestis, and a mouse monoclonal antibody against the capsular antigen fraction 1 (F1) were compared in immunofluorescence (IF) tests. Fluorescent antibody conjugates were prepared from polyclonal antisera to four F1 positive Y. pestis strains; the conjugated antibody to strain A1122 gave the strongest IF staining of F1 positive and F1 negative Y. pestis strains. Indirect assays were rejected in favour of direct assays utilizing polyclonal and monoclonal reagents because the increased background staining reduced the effective contrast of bacterial visualisation. Polyclonal conjugates gave fairly homogeneous staining of Y. pestis bacterial populations, but in monoclonal assays a skew distribution of fluorescence intensity was observed, the majority of bacteria being poorly stained. The proportion of cells stained well by the monoclonal sufficed for easy identification of Y. pestis of the F1 positive phenotype however, and staining was not affected by washing the bacteria or treating them with formaldehyde. Y. pestis strains of the F1 positive genotype reacted with the monoclonal if bacteria were grown at 37 °C but not if the growth temperature was reduced to 25°C thus preventing capsule production. The polyclonal conjugate reacted with bacteria of these strains that had been grown at either temperature. Strains of F1 negative genotype grown at either temperature. Strains of F1 negative genotype grown at either temperature reacted with the polyclonal conjugate but not with the monoclonal. Cross reactions between the polyclonal reagents and Y. enterocolitica biovar 2, serovar O 8 could not be removed by selective absorption; however, the monoclonal antibody gave no cross reaction.

The F1 phenotypic status of bacterial preparations was verified by ELISA measurement of the fraction 1 antigen concentration. Antigen levels for F1 positive and F1 negative phenotypes differed by about three logs for suspensions of Y. pestis harvested from solid media.

The polyclonal and monoclonal direct IF tests applied to spleen and blood smears of laboratory mice infected with Y. pestis were able to differentiate between lethal infection with an F1 positive strain carrying all four classical virulence determinants, an F1 positive vaccine strain, and an F1 negative strain.

Streptococcus pneumoniae strains causing invasive pneumococcal disease (IPD) in the elderly population of England and Wales during the winter of 2003/2004 (1 November 2003 to 30 April 2004) were characterized by serotyping and genotyping in order to determine their population structure in the elderly. Serotyping and multilocus sequence typing (MLST) were carried out on 542 invasive isolates referred to the Respiratory and Systemic Infection Laboratory. Pneumococci were distributed among 32 serotypes and 144 MLST sequence types. A high genetic diversity was observed within the major serotypes. Genetic relatedness varied with regard to serotype. Isolates within serotypes 3, 7F and 8 were the most genetically related whereas serotypes 6A and 19F comprised isolates originating from unrelated ancestors. There was indirect evidence that some pneumococci were derived from clones that had undergone capsular switching in the past. Interestingly one case of IPD was caused by a pneumococcus originating from a clone that had undergone capsular switching from serotype 18C, a serotype included in 7-valent pneumococcal conjugate vaccine (PCV) to serotype 1 (serotype not included in PCV) suggesting that virulent clones with the potential ability to evade PCV existed in the pneumococcal population prior to the routine introduction of this vaccine. Isolates from 28 cases of apparent 23-valent pneumococcal polysaccharide vaccine (PPV) failure were included but there was no evidence of the emergence of particular clones associated with vaccine failures. Longitudinal studies based on serotypic and genetic characterization of pneumococci are fundamental to understanding the impact of both PPV and PCV on the genetic structure of pneumococcal populations.

The objective of this study was to determine the serotype distribution and antibiotic resistance of invasive pneumococcal disease (IPD) strains in children from Lima, Peru, before and after the introduction of the 7-valent pneumococcal conjugate vaccine (PCV7), which was introduced in the national immunisation program on 2009. We conducted a prospective, multicentre, passive surveillance IPD study during 2006–2008 and 2009–2011, before and right after the introduction of PCV7 in Peru. The study was performed in 11 hospitals and five private laboratories in Lima, Peru, in patients <18 years old, with sterile site cultures yielding Streptococcus pneumoniae. In total 159 S. pneumoniae isolates were recovered. There was a decrease in the incidence of IPD in children <2 years old after the introduction of PCV7 (18.4/100 000 vs. 5.1/100 000, P = 0.004). Meningitis cases decreased significantly in the second period (P = 0.036) as well as the overall case fatality rate (P = 0.025), including a decreased case fatality rate of pneumonia (16.3% to 0%, P = 0.04). PCV7 serotypes showed a downward trend. Vaccine-preventable serotypes caused 78.9% of IPD cases, mainly 14, 6B, 5, 19F and 23F. A non-significant increase in erythromycin resistance was reported. Our findings suggest that the introduction of PCV7 led to a significant decrease of IPD in children under 2 years old and in the overall case fatality rate.

Shiga toxin-producing Escherichia coli (STEC) infection can cause serious illness including haemolytic uraemic syndrome. The role of socio-economic status (SES) in differential clinical presentation and exposure to potential risk factors amongst STEC cases has not previously been reported in England. We conducted an observational study using a dataset of all STEC cases identified in England, 2010–2015. Odds ratios for clinical characteristics of cases and foodborne, waterborne and environmental risk factors were estimated using logistic regression, stratified by SES, adjusting for baseline demographic factors. Incidence was higher in the highest SES group compared to the lowest (RR 1.54, 95% CI 1.19–2.00). Odds of Accident and Emergency attendance (OR 1.35, 95% CI 1.10–1.75) and hospitalisation (OR 1.71, 95% CI 1.36–2.15) because of illness were higher in the most disadvantaged compared to the least, suggesting potential lower ascertainment of milder cases or delayed care-seeking behaviour in disadvantaged groups. Advantaged individuals were significantly more likely to report salad/fruit/vegetable/herb consumption (OR 1.59, 95% CI 1.16–2.17), non-UK or UK travel (OR 1.76, 95% CI 1.40–2.27; OR 1.85, 95% CI 1.35–2.56) and environmental exposures (walking in a paddock, OR 1.82, 95% CI 1.22–2.70; soil contact, OR 1.52, 95% CI 2.13–1.09) suggesting other unmeasured risks, such as person-to-person transmission, could be more important in the most disadvantaged group.