Leptospirosis is a zoonosis of worldwide distribution, caused by infection with pathogenic spirochaetes of the genus Leptospira. The wild boar (Sus scrofa), an important hunting species in Europe, seems to play a significant role in the epidemiological cycle of leptospirosis. A total of 101 serum samples from wild boar hunted in Northern Portugal were analysed for leptospiral antibodies detection by microscopic agglutination test. Sera were collected during hunting seasons (2011–2013) and tested with 17 different pathogenic serovars of Leptospira. Antibodies against nine serovars were detected in 66 (65·4%) of these sera. Serovars Tarassovi and Altodouro exhibited the highest seroreactivity rates (23·8% and 16·8%, respectively), followed by Autumnalis (7·9%) and Bratislava (6·9%). Age and district of origin were found to be risk factors for the presence of leptospiral antibodies in contrast to gender. From a One Health perspective, this study revealed that wild boar should be considered as a potential source of leptospirosis dissemination for humans and animal species (domestic and wild) in shared environments, particularly in the Trás-os-Montes region.

Intra-home and kindergarten transmissions were the reported major modes of hand, foot, and mouth disease (HFMD) transmission in preschool children. However, infection at home is not common and 65–80% of cases do not attend preschool. We conducted a matched case-control study to explore the role of public playgrounds in the transmission of HFMD in addition to direct and indirect exposure to HFMD patients. We used 156 hospital source cases and 156 community source controls. Univariate analysis was followed by conditional logistic regression with attributable fraction computed. Adjusted odds ratios were 11·70 [95% confidence interval (CI) 1·26–109·40] for having HFMD cases in the same class, 14·19 (95% CI 3·55–56·74) for having HFMD cases within the 20 nearest neighbourhoods, 6·03 (95% CI 2·84–12·80) for exposure to public playgrounds, 2·13 (95% CI 1·05–4·32) for finger sucking and 0·29 (95% CI 0·11–0·78) for hand washing with soap before meals. The attributable fractions for the first four risk factors were 6·4%, 20·9%, 57·2% and 27·5%, respectively, while the population prevented fraction for hand washing with soap before meals was 18·7%. Based on our findings, hand washing with soap should be advocated. Health education could include topics which underline the precautions which need to be taken and the advice given regarding avoiding the use of public playgrounds during epidemic periods, especially when children have been getting sick.

The incidence of recreational water-associated outbreaks in the United States has significantly increased, driven, at least in part, by outbreaks both caused by Cryptosporidium and associated with treated recreational water venues. Because of the parasite's extreme chlorine tolerance, transmission can occur even in well-maintained treated recreational water venues (e.g. pools) and a focal cryptosporidiosis outbreak can evolve into a community-wide outbreak associated with multiple recreational water venues and settings (e.g. childcare facilities). In August 2004 in Auglaize County, Ohio, multiple cryptosporidiosis cases were identified and anecdotally linked to pool A. Within 5 days of the first case being reported, pool A was hyperchlorinated to achieve 99·9% Cryptosporidium inactivition. A case-control study was launched to epidemiologically ascertain the outbreak source 11 days later. A total of 150 confirmed and probable cases were identified; the temporal distribution of illness onset was peaked, indicating a point-source exposure. Cryptosporidiosis was significantly associated with swimming in pool A (matched odds ratio 121·7, 95% confidence interval 27·4–∞) but not with another venue or setting. The findings of this investigation suggest that proactive implementation of control measures, when increased Cryptosporidium transmission is detected but before an outbreak source is epidemiologically ascertained, might prevent a focal cryptosporidiosis outbreak from evolving into a community-wide outbreak.

To investigate risk factors for sporadic salmonellosis, for each notified case four randomly selected population controls matched for age, sex and geographical region were interviewed via self-administered questionnaire. Conditional logistic regression analysis of 285 matched pairs revealed significant associations for raw ground pork consumption [odds ratio (OR) 6·0, 95% confidence interval (CI) 1·8–20·1], taking antacids (OR 5·8, 95% CI 1·4–24·5), eating meat outside the home (OR 5·7, 95% CI 2·2–14·6) and daily changing or cleaning of dishcloth (OR 2·1, 95% CI 1·2–3·9). Animal contact and ice cream consumption were negatively associated with salmonellosis (OR 0·5, 95% CI 0·2–1 and OR 0·3, 95% CI 0·1–0·6, respectively). S. Typhimurium infections were significantly associated with raw ground pork consumption (OR 16·7, 95% CI 1·4–194·4) and S. Enteritidis infections with having travelled abroad (OR 9·7, 95% CI 2·0–47·3). Raw egg consumption was not a risk factor, substantiating the success of recently implemented national control programmes in the poultry industry. Unexpectedly, hygienic behaviour was more frequently reported by cases, probably because they overestimated their hygiene precautions retrospectively. Although animal contact might enhance human immunocompetence, underreporting of salmonellosis by pet owners could have occurred. Eating raw pork products is the major risk factor for sporadic human S. Typhimurium infections in Lower Saxony.

Meningococcal disease is a rare, rapidly progressing condition which may be difficult to diagnose, disproportionally affects children, and has high morbidity and mortality. Accurate incidence estimates are needed to monitor the effectiveness of vaccination and treatment. We used linked notification, hospital, mortality and birth data for all children of an Australian state (2000–2007) to estimate the incidence of meningococcal disease. A total of 595 cases were notified, 684 cases had a hospital diagnosis, and 26 cases died from meningococcal disease. All deaths were notified, but only 68% (466/684) of hospitalized cases. Of non-notified hospitalized cases with more than one clinical admission, most (90%, 103/114) did not have meningococcal disease recorded as their final diagnosis, consistent with initial ‘false-positive’ hospital meningococcal disease diagnosis. After adjusting for false-positive rates in hospital data, capture–recapture estimation suggested that up to four cases of meningococcal disease may not have been captured in either notification or hospital records. The estimated incidence of meningococcal disease in NSW-born and -resident children aged 0–14 years was 5·1–5·4 cases/100 000 child-years at risk, comparable to international estimates using similar methods, but lower than estimates based on hospital data.

We report on the follow-up and epidemiological study triggered by the isolation of the first vancomycin-resistant Staphylococcus aureus (VRSA) detected in Europe. The patient and 53 close contacts were screened for S. aureus colonization and all isolates recovered were characterized by multiple molecular typing methods. The VRSA remained confined to the infected foot of the patient and was not detected in any of the close contacts. Nasal colonization with S. aureus was detected in 20 subjects, of whom 15 carried methicilin-susceptible isolates with the remaining five harbouring methicilin-resistant S. aureus (MRSA). The majority of the isolates belonged to clones that have been previously shown to be prevalent in Portugal, both in the hospital setting and in the community. Only one isolate, an MRSA, was closely related to the VRSA. Like most of the characterized VRSA isolates from other countries, the VRSA isolated in Portugal belonged to clonal complex (CC) 5. Despite the absence of VRSA dissemination, the recent increase in the incidence of lineages belonging to CC5 in some European countries, including Portugal, may result in more frequent opportunities for the emergence of VRSA.

Faecal samples from 804 cattle and buffaloes and 165 diarrhoeal children of Ismailia province were investigated by an immunochromatographic screening test and PCR to determine prevalences and distributions of Cryptosporidium spp. Results were analysed statistically for clustering of animal and human cases. Cryptosporidium herd prevalence was 73·3% and individual animal prevalence 32·3%. C. parvum was the dominant species in animals (65·7%). Young calves watered with canal or underground water were at particular risk of infection. Detection rates were higher when calves showed diarrhoea, fever and dehydration. Human Cryptosporidium prevalence was 49·1%. C. hominis dominated in humans (60·5%), followed by C. parvum (38·3%). Living in villages, drinking underground water and having contact with animals were risk factors. Cluster analysis revealed differences in the distribution of infections between animals and humans and suggests different transmission dynamics.

The high incidence of norovirus (NoV) infections seems to be related to the emergence of new variants that evolved by genetic drift of the capsid gene. In this work, that represents a first effort to describe the molecular epidemiology of NoV in the northwest of Spain, a total of eight different NoV genotypes (GII.1, GII.3, GII.4, GII.6, GII.7, GII.12, GII.13, GII.14) were detected. The major genotypes observed were GII.4 (45·42%) and GII.14 (34·9%), being detected in all age groups. In addition, and although most of GII.4 sequences belonged to 2006b (7·2%) and 2010 (50·35%) variants, the presence of new NoV variants was observed. Phylogenetic analysis revealed that a high number of GII.4 sequences (35·24%) could be assigned to the newly emerging Sydney 2012 variant, even during late 2010. The high prevalence of NoV GII.14 observed in this study may indicate the emergence of this genotype in Spain.

Infection with hepatitis A virus (HAV) in patient with chronic liver disease (CLD; due to hepatitis B or hepatitis C) may cause severe disease and fulminant liver failure. This study aimed to determine the seroprevalence of HAV antibodies in patients infected with HCV or HBV in Iran (Jahrom city). A total of 159 patients with underlying CLD were recruited between September 2012 and February 2013. Serum samples were collected from each patient and tested for anti-HAV using enzyme-linked immunosorbent assay (ELISA). The overall seroprevalence of total anti-HAV was 79·2%. Patients aged 20–30 years had the lowest (28·3%) anti-HAV seropositivity and those aged >50 years had the highest (95%) seropositivity. The overall prevalence of anti-HAV in patients with chronic HCV and HBV infection was 93·7% and 77·1%, respectively. The anti-HAV seropositivity in liver cirrhosis patients was 100% compared to CLD patients. Because of low HAV immunity in younger CLD patients, vaccination against HAV should be considered.

This study aims to describe in detail the temporal dynamics of E. coli O157 shedding and risk factors for shedding in a grass-fed beef herd. During a 9-month period, 23 beef cows were sampled twice a week (58 sampling points) and E. coli O157 was enumerated from faecal samples. Isolates were screened by PCR for presence of rfbE, stx1 and stx2. The prevalence per sampling day ranged from 0% to 57%. This study demonstrates that many members of the herd were concurrently shedding E. coli O157. Occurrence of rainfall (P < 0·01), feeding silage (P < 0·01) and lactating (P < 0·01) were found to be predictors of shedding. Moving cattle to a new paddock had a negative effect on shedding. This approach, based on short-interval sampling, confirms the known variability of shedding within a herd and highlights that high shedding events are rare.

This study characterized Neisseria meningitidis serogroup C strains in China in order to establish their genetic relatedness and describe the use of multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) to provide useful epidemiological information. A total of 215 N. meningitidis serogroup C strains, obtained from 2003 to 2012 in China, were characterized by MLVA with different published schemes as well as multilocus sequence typing. (i) Based on the MLVA scheme with a combination of five highly variable loci, 203 genotypes were identified; this level of discrimination supports its use for resolving closely related isolates. (ii) Based on a combination of ten low variable loci, clear phylogenetic relationships were established within sequence type complexes. In addition, there was evidence of microevolution of VNTR loci over the decade as strain lineages spread from Anhui to other provinces, the more distant the provinces from Anhui, the higher the genetic variation.

Environmental samples are a cost-saving and easy-to-use approach to diagnose paratuberculosis at the herd level. Detailed knowledge concerning its uncertainties in herds with a low prevalence of Mycobacterium avium spp. paratuberculosis (MAP) is required to design sampling strategies in control programmes. This study aimed to calculate a threshold level of the apparent within-herd prevalence (WHPapp) as determined by individual mycobacterial cultivation (faecal culture; FC) of all cows thus allowing the detection of a herd as MAP-positive at a certain probability level (P d). Out of 200 environmental samples taken twice from five predefined locations in a barn, 25 were positive by FC and 60 were positive by a quantitative real-time PCR method (qPCR). A logistic regression model was used to calculate the WHPapp threshold of detection. For 50% P d, a WHPapp threshold of 2·9% was calculated for the combination of three samples (milking area, main cow alleyways, holding pen) tested simultaneously both by FC and qPCR. The threshold increased to 6·2% for 90% P d. Repeated environmental sampling did not reduce the WHPapp threshold. Depending on the particular needs for prevalence estimation or in control programmes (single or repeated sampling) the provided WHPapp thresholds at different P d will enable decisions to be made about various sampling strategies.

Multi-locus variable number tandem repeat analysis (MLVA) is used in clinical and reference laboratories for subtyping verocytotoxin-producing Escherichia coli O157 (VTEC O157). However, as yet there is no common allelic or profile nomenclature to enable laboratories to easily compare data. In this study, we carried out an inter-laboratory comparison of an eight-loci MLVA scheme using a set of 67 isolates of VTEC O157. We found all but two isolates were identical in profile in the two laboratories, and repeat units were homogeneous in size but some were incomplete. A subset of the isolates (n = 17) were sequenced to determine the actual copy number of representative alleles, thereby enabling alleles to be named according to international consensus guidelines. This work has enabled us to realize the potential of MLVA as a portable, highly discriminatory and convenient subtyping method.

Escherichia coli O157:H7 strains (n = 33) recovered from different food sources in Egypt were characterized using molecular assays to identify strain genotypes associated with various levels of pathogenic potential. Genotypic characterization included: lineage-specific polymorphism assay (LSPA-6), Shiga-toxin-encoding bacteriophage insertion site assay (SBI), clade 8 typing, Tir (A255 T) polymorphism, and variant analysis of Shiga toxin 2 gene (Stx2a and Stx2c), and anti-terminator Q genes (Q933 and Q21). Genotypes LI/II (76%), SBI 1 (60·6%), clade 8 (69·7%), Tir (255 T) (72·7%) and Stx2c (45·5%) were found to be significantly more frequent compared to other genetic markers in the strains analysed. Multivariable analysis revealed a significant association between LPSA-6 and clade types as well as Tir (A255 T). To the best of our knowledge, this is the first study to report the characterization of these genetic markers in E. coli O157:H7 strains in the Middle East and Africa.

Mycoplasma pneumoniae outbreaks cause increased use of macrolides and tetracyclines. We aimed to investigate whether drug use data, in addition to laboratory data, could improve understanding of the spread of M. pneumoniae epidemics. Number of users of Mycoplasma antibiotics (erythromycin, doxycycline, clarithromycin) per week and county of residence in an indicator age group (6–12 years) was retrieved from the Norwegian prescription database for the epidemic season 2011–2012 and compared to non-epidemic seasons. In 2011, increased use of Mycoplasma antibiotics was first observed in September on the west coast of Norway. The Norwegian laboratory-based surveillance system showed the first increase in positive tests in August 2011 and an epidemic was announced on 25 October 2011. At that time the use of Mycoplasma antibiotics had already exceeded three times the use in non-epidemic periods. Data for three counties from the regional microbiological laboratories showed that the increase in number of positive samples coincided in time with the increase in prescription data. Laboratory data cannot accurately determine the extent of an epidemic, and drug use data cannot identify the cause. Establishing a systematic interaction between the two monitoring systems will enhance surveillance and probably contribute to improved infection control and prudent antibiotic prescribing.

This study developed a fast and high throughput dot-blot technique to evaluate the presence of Entamoeba in stool samples (n = 643) followed by a PCR-based method to validate and differentiate the two species E. histolytica and E. dispar. The prevalence rate of the parasite has been detected in a cross-sectional study carried out in the population of the Eastern and Northern parts of India. Of the various demographic features, prevalence was highest in the monsoon season (P = 0·017), in the <15 years age group (P = 0·015). In HIV-positive individuals, the prevalence rate was significantly high (P = 0·008) in patients with a CD4 cell count <200 as well as in patients without antiretroviral therapy (ART) (P = 0·011). Our analysis further confirmed that risk factors such as toilet facilities, living conditions, hygienic practices, drinking water source, occupation and level of education are important predictors as they were found to contribute significantly in the prevalence of the parasite.

Information on the risk factors for community-associated skin and soft-tissue infections (SSTIs) due to methicillin-resistant Staphylococcus aureus in Asian populations is scarce. To this end we performed a case-control study of patients treated at two hospital-affiliated outpatient clinics in Taiwan to determine potential risk factors for MRSA SSTIs. S. aureus was isolated from 39 of 100 eligible patients, and 74% were MRSA. Apart from resistance to clindamycin and erythromycin, most MRSA isolates were susceptible to appropriate antimicrobials. The significant risk factors identified by multivariate analysis for MRSA SSTIs were male gender (P = 0·09), nasal carriage of MRSA (P = 0·02), exposure to an individual who had surgery within a year before infection (P = 0·02), and antibiotic treatment for SSTI in the year before infection (P = 0·04). The identification of such factors may assist provision of appropriate treatment to patients with suspected S. aureus SSTIs particularly in Taiwan.

Q fever patients are often reported to experience a long-term impaired health status, including fatigue, which can persist for many years. During the large Q fever epidemic in The Netherlands, many patients with a laboratory-confirmed Coxiella burnetii infection were not notified as acute Q fever because they did not fulfil the clinical criteria of the acute Q fever case definition (fever, pneumonia and/or hepatitis). Our study assessed and compared the long-term health status of notified and non-notified Q fever patients at 4 years after onset of illness, using the Nijmegen Clinical Screening Instrument (NCSI). The study included 448 notified and 193 non-notified Q fever patients. The most severely affected subdomain in both patient groups was ‘Fatigue’ (50·5% of the notified and 54·6% of the non-notified patients had severe fatigue). Long-term health status did not differ significantly between the notified and non-notified patient groups, and patients scored worse on all subdomains compared to a healthy reference group. Our findings suggest that the magnitude of the 2007–2009 Q fever outbreak in The Netherlands was underestimated when only notified patients according to the European Union case definition are considered.

Serotype 1 Streptococcus pneumoniae is a cause of invasive pneumococcal disease (IPD) worldwide and has been associated with IPD outbreaks, while carriage is rarely detected in healthy adults or children. This study details an Australian multi-state and territory outbreak of serotype 1 S. pneumoniae IPD between 2010 and 2012. Molecular characterization demonstrated the outbreak was largely due to the clonal expansion of sequence type 306, MLVA type 261 S. pneumoniae serotype 1.