1. A historical survey is given of work upon the Eijkman method.

2. A total of 104 water samples were examined by methods I and III. These yielded 602 cultures obtained from the 44 and 37° C. presumptive series, and of these 357 were 44° C.-positive. The only types capable of fermenting lactose at 44° C. were Bact. coli type I and Irregular type II, with the addition of three cultures of intermediate type I and two of aerogenes type I.

3. The relative efficiencies in the detection of Bact. coli of the 44° C. method by direct inoculation and of the 37° C. presumptive test with plating and differentiation were compared. Out of 104 samples, sixty-eight were shown to contain faecal Bact. coli, and in 19% of these the 44° C. method detected Bact. coli when it was not recovered at 37° C. The converse was true in only 13% of samples.

4. The results obtained by other workers in regard to the specificity of incubation at 44° C. and to the various methods hitherto proposed for employing it have been collected and discussed; technical advantages and disadvantages have also been considered.

5. It is concluded: (1) That false positives at 44° C. are a negligible quantity for practical purposes. (2) That the best all-round method to use is Wilson's method IV. (3) That 48 hr. incubation should be given, but that 24 hr. is sufficient when rapidity is desired. (4) That the test should be officially recognized and standards laid down for use with it.

6. Cellobiose was found to be inferior to citrate for the identification of the I.A.C. group.

1. Rats taken from mixed stock diet and fed synthetic diet containing an excess of magnesium carbonate developed urinary calculi in a large number of cases.

2. The incidence of calculi was much higher in the females.

3. Addition of excess of calcium carbonate to the diet prevented the formation of calculi.

4. Rats previously fed for a short time on the synthetic diet and normal salt mixture did not develop calculi, but suffered from severe catharsis.

5. Rats on mixed stock diet had a more acid caecum and colon than those on synthetic diet.

6. No significant alterations were found in the Ca, Mg or inorganic P of the blood on any of the diets.

7. Extra magnesium, or calcium, or sodium carbonate lowered the urinary phosphate excretion.

8. Extra magnesium or calcium had little effect upon the urinary excretion of the other, but tended in each case to lower the output.

9. Changes in the sex organs brought about by excessive intake of magnesium are described.

The writer wishes to thank Miss V. E. Leader for her watchful care of the rats.

Avian influenza (AI) is an important disease that has significant implications for animal and human health. High pathogenicity AI (HPAI) has emerged in consecutive seasons within the UK to cause the largest outbreaks recorded. Statutory measures to control outbreaks of AI virus (AIV) at poultry farms involve disposal of all birds on infected premises. Understanding of the timing of incursions into the UK could facilitate decisions on improved responses. During the autumnal migration and wintering period (autumn 2019– spring 2020), three active sampling approaches were trialled for wild bird species considered likely to be involved in captive AI outbreaks with retrospective laboratory testing undertaken to define the presence of AIV.

Faecal sampling of birds (n = 594) caught during routine and responsive mist net sampling failed to detect AIV. Cloacal sampling of hunter-harvested waterfowl (n = 146) detected seven positive samples from three species with the earliest detection on the 17 October 2020. Statutory sampling first detected AIV in wild and captive birds on 3 November 2020. We conclude that hunter sourced sampling of waterfowl presents an opportunity to detect AI within the UK in advance of outbreaks on poultry farms and allow for early intervention measures to protect the national poultry flock.

The use of antimicrobials in food-producing animals can lead to increased bacterial resistance. Important information to address this problem can be provided by monitoring antimicrobial resistance (AMR) in foodborne pathogens. As part of preliminary activities for the implementation of AMR surveillance in Brazil, a nationwide survey on AMR in Salmonella enterica isolates from poultry meat was conducted. The survey evaluated 146 Salmonella isolates from poultry meat in 2014, and 163 isolates obtained in 2017. Minimal inhibitory concentrations of 13 antimicrobials were determined by broth microdilution, and isolates were assigned to serotypes by automated ribotyping. High resistance rates were found in 2014 and 2017, in particular to nalidixic acid (84/146, 57.5% and 141/163, 86.5%, respectively), ampicillin (82/146, 56.2% and 125/163, 76.7%), cefotaxime (76/146, 52.1% and 124/163, 76.1%), ceftazidime (73/146, 50.0% and 124/163, 76.1%), ciprofloxacin (83/146, 56.9% and 145/163, 89.0%) and tetracycline (88/146, 60.3% and 135/163, 82.8%). There was a significant increase in resistance to these antibiotics in the second survey period. Salmonella ser. Heidelberg and Salmonella ser. Minnesota were the main serotypes expressing resistance to these antimicrobials. Multidrug resistance was found in 50.7% (74/146) of the isolates from 2014, and in 77.3% (126/163) of isolates from 2017 (P < 0.05). None of the isolates was resistant to azithromycin or meropenem. These findings indicate high and increasing rates of resistance among Salmonella from poultry meat in Brazil, mainly associated with Salmonella ser. Heidelberg and Salmonella ser. Minnesota, stressing the importance of continuous monitoring of AMR in the poultry chain.

We investigated a measles outbreak in the Jerusalem district in 2007–2008 (992 cases). Most cases (72·6%) were aged <15 years, 42·9% aged <5 years, and 12·8% were infants aged <1 year. The peak incidence rate was in infants aged 6–12 months (916·2/100 000). This represents a significant shift from former outbreaks in 2003–2004, where the peak incidence was in the 1–4 years age group. Of children aged <5 years the proportion aged 6–12 months tripled (7·7% vs. 25·6%). In a case-control study (74 cases, 148 controls) children who developed measles were less likely to be registered in a well-baby clinic and had lower overall immunization coverage. The differences in proportions for registration, DTaP3 and MMR1 coverage were 35·1%, 48·6% and 80·8%, respectively (all P<0·001). Rising birth order of cases and their siblings was associated with non-registration and non-compliance with MMR immunization. The vulnerability of young infants and the risk markers noted above should be taken into account in planning intervention programmes.

Maximum entropy ecological niche modelling and spatial scan statistic were utilized to predict the geographical range and to investigate clusters of infections for Neospora caninum and Coxiella burnetii in dairy cattle farms in Catalonia, northeastern Spain, using the Maxent and SaTScan programs, respectively. The geographical distribution of Neospora and Coxiella with the highest level of probability (P > 0·60) covers central Catalonia and spreads towards northeastern Catalonia which contains a high concentration of dairy cattle farms. The most important environmental factor that contributed to the ecological niche modelling was precipitation of driest month followed by elevation. Significant clusters (P < 0·001) were detected for Neospora and Coxiella infections in the western and eastern regions of Catalonia, respectively.

Bloodstream infection (BSI) is a serious complication in immunocompromised hosts. This study compares epidemiological, clinical and microbiological characteristics of BSI among children with haematological malignancies (HM) and solid tumours (ST). The study was conducted from October 2012 through to November 2015 at a referral hospital for cancer care and included the first BSI episode detected in 210 patients aged 18 years or less. BSI cases were prospectively detected by daily laboratory-based surveillance. The Centers for Disease Control and Prevention definitions for primary or secondary BSI were used. A higher proportion of use of corticosteroids (P = 0.02), chemotherapy (P = 0.01) and antibiotics (P = 0.05) before the BSI diagnosis; as well as of neutropenia (P < 0.001) and mucositis (P < 0.001) at the time of BSI diagnosis was observed in patients with HM than with ST. Previous surgical procedures (P = 0.03), mechanical ventilation (P = 0.01) and bed confinement (P < 0.001) were more frequent among children with ST. The frequency of use of temporary (P = 0.01) and implanted vascular lines (P < 0.01) was significantly higher in children with ST than with HM while the tunnelled line (P = 0.01) use was more frequent in children with HM as compared to ST. Most (n = 181) BSI cases were primary BSI. BSI associated with a tunnelled catheter was more frequent in children with HM (P < 0.01), whereas BSI associated with an implanted (P < 0.01) or temporary central line (P < 0.02) was more common in patients with ST. BSI associated with mucosal barrier injury was more frequent (P = 0.01) in children with HM. Indication for intensive care was more frequent in children (P = 0.05) with ST. Mortality ratio was similar in children with ST and HM, and length of hospital stay after BSI was higher in patients with HM than with ST (median of 19 vs. 13 days; P = 0.02). Infection caused by Gram-negative bacteria (P = 0.04) and polymicrobial infections (P = 0.05) due to Gram-positive cocci plus fungus was more common in patients with HM. These findings suggest that the characteristics of BSI acquisition and mortality can be cancer-specific.

Inclusion of live varicella vaccine in the routine occupational health vaccination schedule requires knowledge of the natural immunity to varicella zoster virus (VZV) among high-risk occupations. This study aims were to evaluate VZV antibody positivity among health-care workers (HCWs) and day-care-centre workers (DCWs) and to assess its association with potential risk factors. Three groups of workers were tested for VZV antibody positivity: hospitals and community clinic HCWs (n=335), DCWs (n=117) and blue-collar workers as controls (n=121). The total VZV antibody positivity was 94·4%. There was no significant difference in VZV antibody positivity among study groups. DCWs had the lowest VZV seroprevalence (90·9%, 95% CI 85·7–96·1) and controls the highest (96·6%, 95% CI 93·2–99·9). This high VZV antibody positivity suggests that no special occupational measures are indicated in health-care or day-care occupational settings in Israel. On-going monitoring of the natural immunity to VZV is necessary to detect trends over time.

As acute infectious pneumonia, the coronavirus disease-2019 (COVID-19) has created unique challenges for each nation and region. Both India and the United States (US) have experienced a second outbreak, resulting in a severe disease burden. The study aimed to develop optimal models to predict the daily new cases, in order to help to develop public health strategies. The autoregressive integrated moving average (ARIMA) models, generalised regression neural network (GRNN) models, ARIMA–GRNN hybrid model and exponential smoothing (ES) model were used to fit the daily new cases. The performances were evaluated by minimum mean absolute per cent error (MAPE). The predictive value with ARIMA (3, 1, 3) (1, 1, 1)14 model was closest to the actual value in India, while the ARIMA–GRNN presented a better performance in the US. According to the models, the number of daily new COVID-19 cases in India continued to decrease after 27 May 2021. In conclusion, the ARIMA model presented to be the best-fit model in forecasting daily COVID-19 new cases in India, and the ARIMA–GRNN hybrid model had the best prediction performance in the US. The appropriate model should be selected for different regions in predicting daily new cases. The results can shed light on understanding the trends of the outbreak and giving ideas of the epidemiological stage of these regions.

Parvovirus B19 (B19V) infection during pregnancy may have serious consequences like fetal anaemia, hydrops fetalis, and fetal loss. Since epidemiological data on B19V infection are generally lacking in Sudan, the current study aimed to determine the seroprevalence of B19V in Sudanese pregnant women. Five hundred women, attending antenatal clinics in Khartoum state between November 2008 and March 2009, were enrolled and screened for B19V IgG and IgM antibodies by enzyme immunoassays. The study revealed a B19V IgG seroprevalence of 61·4%, with one subject positive for IgM. B19V DNA was not detected by PCR in any of the tested individuals. B19V IgG seroprevalence was significantly correlated with multigravidity (P = 0·046). Our data showed that B19V infection is prevalent in Sudan and we recommend further studies in Sudanese women, particularly in those with complications and adverse outcomes of pregnancy.

Death certificate reports and laboratory-confirmed influenza deaths probably underestimate paediatric deaths attributable to influenza. Using US mortality data for persons aged <18 years who died during 28 September 2003 to 2 October 2010, we estimated influenza-attributable deaths using a generalized linear regression model based on seasonal covariates, influenza-certified deaths (deaths for which influenza was a reported cause of death), and occurrence during the 2009 pandemic period. Of 32 783 paediatric deaths in the death categories examined, 853 (3%) were influenza-certified. The estimated number of influenza-attributable deaths over the study period was 1·8 [95% confidence interval (CI) 1·3–2·8] times higher than the number of influenza-certified deaths. Influenza-attributable deaths were 2·1 (95% CI 1·5–3·4) times higher than influenza-certified deaths during the non-pandemic period and 1·1 (95% CI 1·0–1·8) times higher during the pandemic. Overall, US paediatric deaths attributable to influenza were almost twice the number reported by death certificate codes in the seasons prior to the 2009 pandemic.

The aim of the European Sero-Epidemiology Network 2 (ESEN2) project was to estimate age-specific seroprevalence for a number of vaccine-preventable diseases in Europe. To achieve this serosurveys were collected by 22 national laboratories. To adjust for a variety of laboratory methods and assays, all quantitative results were transformed to a reference laboratory's units and were then classified as positive or negative to obtain age-specific seroprevalence. The aim of this study was to assess the value of standardization by comparing the crude and standardized seroprevalence estimates. Seroprevalence was estimated for measles, mumps, rubella, diphtheria, varicella zoster and hepatitis A virus (HAV) and compared before and after serological results had been standardized. The results showed that if no such adjustment had taken place, seroprevalence would have differed by an average of 3·2% (95% bootstrap interval 2·9–3·6) although this percentage varied substantially by antigen. These differences were as high as 16% for some serosurveys (HAV) which means that standardization could have a considerable impact on seroprevalence estimates and should be considered when comparing serosurveys performed in different laboratories using different assay methods.

Aimed at assessing the circulation of Mycobacterium tuberculosis in a highly endemic prison, this 13-month prospective study was performed on strains isolated from tuberculosis (TB) cases detected passively and actively. We used X-ray screening of newly admitted inmates and mass screening at the beginning of the study and again 1 year later. Of the 94 strains genotyped by restriction fragment-length polymorphism, 79 (84·0%) belonged to one of the 12 identified clusters (2–21 strains each), including two main clusters (18 and 21 cases, respectively). A history of TB treatment was reported in 22/79 (27·8%) clustered cases. Time–space distribution of clustered cases was predominantly consistent with transmission, in micro-epidemics. Given the dominant pattern of exogenous infection and the extensive strain circulation, effective TB control should emphasize reduction of overcrowding and improvement of environmental measures as a complement to detection and treatment of cases.

Chronic irritation is a recognized factor in the causation of malignant epithelial growths and it would not, a priori, be surprising if an association were found to exist between pneumokoniosis and pulmonary cancer. With the notable and somewhat questionable exceptions of the observations upon Schneeberg and Joachimstal miners there has, however, been little to support this view. Nevertheless certain observations of ours (Dible, 1934) have urged us to examine the relationship of silicosis to malignancy in a more specific and comprehensive fashion than has hitherto been achieved. This paper records the results of our investigations, which it is thought may bear upon the aetiology of the condition.

Clean clothes in the staff dressing rooms were heavily contaminated with bacteria, mainly Bacillus sp., but Staphylococcus aureus were found on 14% and Clostridium sp. on 10% of the garments examined. A comparison of the occurrence of Staph. aureus on shirts worn by staff in wards and operating departments showed ward shirts to be contaminated more heavily and with more strains. Examination of sterile gowns worn by surgeons showed that 70% were contaminated with Staph. aureus after operation. Of the strains isolated 31% were identical with those carried by the surgeon or by the patient operated on, but for the remainder no source could be found.

Enterotoxigenic F4+ Escherichia coli can colonize the intestine of pigs and cause diarrhoea. Our primary goal was to find a discriminant rule to discriminate between F4+ E. coli shedding profiles as this may reflect differences in the infectiousness of pigs. Our secondary goal was to find a discriminant rule to discriminate between diarrhoeic and non-diarrhoeic pigs. Repeated measurements (bacterial shedding and percentage dry matter of faeces) were taken of 74 weaned pigs that were infected experimentally with F4+ E. coli. These measurements were summarized into two new variables by means of a principal components analysis. Discriminant rules were derived based on these summary variables by fitting a mixture of normal distributions. Finally, the association between the classifications (as derived from the discriminant rules) and the occurrence in the pigs of the F4 receptor, an adhesion site for F4+ E. coli, was studied. We found that only the classification based on bacterial shedding allowed us to distinguish two significantly different groups of pigs (high and low shedders). Presence of the F4 receptor was associated strongly with pigs being high shedders.

In previous work in this laboratory, Mycoplasma suipneumoniae was recovered in liquid medium from 13 % of individual cases and 18 % of outbreaks of enzootic pneumonia in pigs. In the work now described, however, these recovery rates, when judged by the same criteria, were 45 and 75 %, respectively. As there was evidence to suggest that this second series of pneumonic cases was less suitable for cultural examination than the first series, some of the other factors that might have improved the recovery rate were investigated.

Some improvement was probably achieved by inoculating the liquid medium with three or four different dilutions of pneumonic tissue, each dilution always being in duplicate, and by incubating the inoculated tubes for over 3 weeks before discarding them.

A second advantage could have derived from the fact that all batches of liquid medium were tested for their ability to support the growth of M. suipneumoniae before being used to culture field material.

The effect of varying one constituent at a time was observed in controlled experiments: different batches of pig sera had a marked, variable effect on the growth of both M. suipneumoniae and Mycoplasma hyorhinis; medium made with purchased Hartley's broth was found to be superior to medium incorporating broth made in this laboratory, more so for the growth of M. suipneumoniae than M. hyorhinis; the incorporation of yeast extract made in this laboratory gave a marginal advantage for the growth of M. hyorhinis; and both mycoplasmas grew equally well in medium containing or lacking thallium acetate.

Some batches of medium were, by chance, markedly selective for the growth of M. suipneumoniae compared with M. hyorhinis. As the full reasons for this were not known, attempts were made to develop selective media in a more direct way. One such medium contained 5% pig serum and 15% horse serum, and a second was of similar composition, except that the pig serum used inhibited preferentially the growth of M. hyorhinis compared with M. suipneumoniae. Both media markedly favoured the growth of M. suipneumoniae when tested separately with cultures of M. suipneumoniae and M. hyorhinis. The second medium yielded M. suipneumoniae when inoculated with a 10-1 dilution of a culture of M. suipneumoniae and a 10-2 dilution of a culture of M. hyorhinis, whereas a standard batch, of liquid medium, similarly inoculated with M. suipneumoniae did not yield this mycoplasma until the M. hyorhinis culture included in the inoculum was diluted to 10-6.

Both selective media, when tested on a small number of field cases, gave improved isolations of M. suipneumoniae compared with the routine batches of liquid medium used initially.

Considerable difficulty was experienced in producing a sufficiently high level of antibodies to M. hyorhinis in pig sera and to M. suipneumoniae in rabbit sera. This exacerbated the problem of isolating and identifying M. suipneumoniae from field cases of enzootic pneumonia by this cultural method.

This study aimed to investigate the clinical characteristics and to analyse the epidemiological features of coronavirus disease 2019 (COVID-19) patients during convalescence. In this study, we enrolled 71 confirmed cases of COVID-19 who were discharged from hospital and transferred to isolation wards from 6 February to 26 March 2020. They were all employees of Zhongnan Hospital of Wuhan University or their family members of which three cases were <18 years of age. Clinical data were collected and analysed statistically. Forty-one cases (41/71, 57.7%) comprised medical faculty, young and middle-aged patients (aged ⩽60 years) accounted for 81.7% (58/71). The average isolation time period for all adult patients was 13.8 ± 6.1 days. During convalescence, RNA detection results of 35.2% patients (25/71) turned from negative to positive. The longest RNA reversed phase time was 7 days. In all, 52.9% of adult patients (36/68) had no obvious clinical symptoms, and the remaining ones had mild and non-specific clinical symptoms (e.g. cough, sputum, sore throat, disorders of the gastrointestinal tract etc.). Chest CT signs in 89.7% of adult patients (61/68) gradually improved, and in the others, the lesions were eventually absorbed and improved after short-term repeated progression. The main chest CT manifestations of adult patients were normal, GGO or fibre streak shadow, and six patients (8.8%) had extrapulmonary manifestations, but there was no significant correlation with RNA detection results (r = −0.008, P > 0.05). The drug treatment was mainly symptomatic support therapy, and antibiotics and antiviral drugs were ineffective. It is necessary to re-evaluate the isolation time and standard to terminate isolation for discharged COVID-19 patients.

The epidemiological, clinical and therapeutic features of HIV disease diagnosed in 41 immigrants from outside of the European Union (EU), were compared with those of 123 Italian and EU patients, in a cross-sectional case-control study, with individuals matched according to age and gender. In total 4·15% of our patients came from outside of the EU (51·2% of them from subsaharan Africa), with a proportional predominance of females, and heterosexual and perinatal transmission of HIV disease (P < 0·0001 and P < 0·02, respectively). Compared with Italian and EU subjects, patients coming from abroad had a shorter duration of known HIV infection (P < 0·001), but only some subjects were aware of their HIV disease prior to immigration, or acquired HIV infection only after coming to Italy (14·6% and 12·2%, respectively). No cases of HIV-2 infection or co-infection were detected in either study group. Compared with controls, patients coming from outside of the EU had a comparable clinical and immunological status, and had similar antiretroviral therapy, which was administered earlier (P < 0·0001), and proved better tolerated (P < 0·04), than in Italian and EU subjects. The apparently more limited virological response (as expressed by a higher mean plasma viral load, and a lower rate of viral suppression at the last visit; P < 0·001 and P < 0·05, respectively), was probably a bias due to the shorter mean overall follow-up time (P < 0·0001), and the shorter mean duration of antiretroviral treatment (11·1±2·2 months of immigrants vs. 16·2±6·7 months of controls; P < 0·0001).

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.