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Objectives To determine spatial clustering and risk factors for occurrence and intensity of infection for soil-transmitted helminthiasis (STH), namely Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis and hookworms in a Venezuelan rural community. Methods MIF-fixed faecal samples were individually collected for STH testing. The Getis–Ord statistic was used to determine significant STH clustering within 25/50/100 m radiuses around houses. Individual- and house-level factors associated with STH occurrence and intensity of infection were determined using generalized estimating equations. Results Significant clusters of “wormy” houses for one or multiple parasites were found at distances of 25–50 m around 13 houses. Risk factors differed between occurrence and intensity of infection. Overcrowding in the house increased occurrence of S. stercoralis, T. trichiura and hookworm infections, while poor housing conditions increased A. lumbricoides infection risk. Overcrowding, poor faecal disposal system, economic dependency and lack of basic services differentially influenced the STHs. The “wormy” houses were mainly those built with waste materials, under economic dependency and lacking indoor water supply. Conclusions STH distribution in a community is clustered, with significant hotspots of STH occurrence and intensity of infection and different associated risk factors. Targeting the “wormy” houses is expected to affect STH morbidity more efficiently.

Suboptimal animal welfare may affect natural immunity, rendering animals more susceptible to environmentally conditioned diseases, including those requiring antimicrobial treatment, which may promote antimicrobial resistance (AMR) in bacterial populations. Herewith, we tested the hypothesis that conventionally raised turkeys have higher levels of AMR in indicator Escherichia coli bacteria, but lower levels of natural immunity, as compared to turkeys reared under organic conditions. Litter and serum samples were collected from 28 conventional and 4 organic turkey farms: E. coli isolates from litter were tested for resistance to 14 antimicrobials, while 3 parameters of natural immunity (i.e., lysozyme, hemolytic complement levels, and serum bactericidal activity) were assessed in the sera. Resistant E. coli isolates were identified in both conventional and organic farms but generally more frequently in conventional farms. High rates of resistance to ampicillin (96%), tetracycline (95%), streptomycin (82%), sulfamethoxazole (80%), ciprofloxacin (73%), and trimethoprim (71%), as well as high rates of multiresistance, were observed in conventional farms. Organically raised turkeys had significantly higher levels of lysozyme and serum bactericidal activity than conventional turkeys, and these levels were also higher in turkeys housed in farms where AMR frequency was lower. Findings support the hypothesis that conventional farming conditions may affect turkeys' natural immunity, rendering the animals more susceptible to environmentally conditioned diseases requiring antimicrobial treatment, which would in turn promote AMR. Reducing AMR in turkey farming is therefore more likely to be successful when considering animal welfare as an option to reduce the need of antimicrobial use.

ObjectivesWhile colon cancer (CC) risk is associated with several lifestyle-related factors, including physical inactivity, smoking and diet, the contribution of occupation to CC morbidity remains largely unclear. Growing evidence indicates that gastrointestinal infections like salmonellosis could contribute to CC development. We performed a nationwide registry study to assess potential associations between occupation (history) and CC, including also those occupations with known increased exposure to gastrointestinal pathogens like Salmonella.MethodsPerson-level occupational data for all residents in The Netherlands were linked to CC diagnosis data. Differences in the incidence of (overall, proximal and distal) CC among occupational sectors and risk groups were tested for significance by calculating standardised incidence ratios (SIRs) with 95% CIs using the general population as reference group. Effects of gender, age, exposure duration and latency were also assessed.ResultsSignificant differences in CC incidence were observed only for a few occupational sectors, including the manufacturing of rubber and plastics, machinery and leather, the printing sector and the information service sector (SIRs 1.06–1.88). No elevated risk of CC was observed among people with increased salmonellosis risk through occupational exposure to live animals, manure or among those working in the sale of animal-derived food products (SIRs 0.93–0.95, 0.81–0.95 and 0.93–1.09 for overall, proximal and distal CC, respectively).ConclusionsThe results of this study suggest that occupation in itself provides a relatively small contribution to CC incidence. This is consistent with previous studies where a similar degree of variation in risk estimates was observed. The lack of an association with the high-risk occupations for salmonellosis might be due to higher levels of physical activity, a known protective factor for CC and other diseases, of people working in the agricultural sector, which might outweigh the potential Salmonella-associated risk of CC.

Highlights • National burden of disease studies are needed to define food safety priorities. • Initiatives to support capacity building and collaboration are emerging. • Novel methodologies provide opportunities to improve data collection for disease burden estimation in low-resource settings., National burden of foodborne disease (FBD) studies are essential to establish food safety as a public health priority, rank diseases, and inform interventions. In recent years, various countries have taken steps to implement them. Despite progress, the current burden of disease landscape remains scattered, and researchers struggle to translate findings to input for policy. We describe the current knowledge base on burden of FBDs, highlight examples of well-established studies, and how results have been used for decision-making. We discuss challenges in estimating burden of FBD in low-resource settings, and the experience and opportunities deriving from a large-scale research project in these settings. Lastly, we highlight the role of international organizations and initiatives in supporting countries to develop capacity and conduct studies.

Antimicrobials are commonly used in conventional livestock production and manure is widely applied to agricultural lands as fertilizer. This practice raises questions regarding the effects of fertilization on (i) soil microbiota composition and (ii) spread of antimicrobials and antimicrobial resistance (AMR) in the environment. This study was conducted in a high-density farming area of Northern Italy and aimed at assessing the impact of (dairy cattle, chickens and swine) manure application on soil microbiome, antimicrobial concentrations and antimicrobial resistance gene (ARG) abundance. We found the microbial community composition in manure to be different and less diverse than in soil, with manure application altering only marginally the soil microbiome. Exceptions were the phyla Firmicutes, Tenericutes and Cloacimonetes, which significantly enriched in fertilized soil. Of the antimicrobials investigated, only flumequine concentrations increased after manure application, albeit non-significantly. ARGs were more abundant in manure, with ermA, ermB, blaOXA-1 and oqxA being significantly enriched in fertilized soil. Positive correlations between oqxA and qnrS abundances and flumequine concentrations were observed, together with the co-occurrence of some ARGs and microbial taxa (e.g. oqxA correlated with Acidobacteria and Gemmatimonadetes). This study showed that manure application has little effect on soil microbiome but may contribute to the dissemination of specific ARGs into the environment. Moreover, flumequine residues seem to enhance the emergence of oqxA and qnrS in soil.

Laboratory data increasingly suggest that Salmonella infection may contribute to colon cancer (CC) development. Here, we examined epidemiologically the potential risk of CC associated with salmonellosis in the human population. We conducted a population-based cohort study using four health registries in Denmark. Person-level demographic data of all residents were linked to laboratory-confirmed non-typhoidal salmonellosis and to CC diagnoses in 1994–2016. Hazard ratios (HRs) for CC (overall/proximal/distal) associated with reported salmonellosis were estimated using Cox proportional hazard models. Potential effects of serovar, age, sex, inflammatory bowel disease and follow-up time post-infection were also assessed. We found no increased risk of CC ≥1 year post-infection (HR 0.99; 95% confidence interval (CI) 0.88–1.13). When stratifying by serovar, there was a significantly increased risk of proximal CC ≥1 year post-infection with serovars other than Enteritidis and Typhimurium (HR 1.40; 95% CI 1.03–1.90). CC risk was significantly increased in the first year post-infection (HR 2.08; 95% CI 1.48–2.93). The association between salmonellosis and CC in the first year post-infection can be explained by increased stool testing around the time of CC diagnosis. The association between proximal CC and non-Enteritidis/non-Typhimurium serovars is unclear and warrants further investigation. Overall, this study provides epidemiological evidence that notified Salmonella infections do not contribute significantly to CC risk in the studied population.

Salmonella Infantis is a poultry-adapted Salmonella enterica serovar that is increasingly reported in broilers and is also regularly identified among human salmonellosis cases. An emerging S. Infantis mega-plasmid (pESI), carrying fitness, virulence and antimicrobial resistance genes, is also increasingly found. We investigated the prevalence, genetic characteristics and risk factors for (pESI-carrying) S. Infantis in broilers. Faecal samples from 379 broiler flocks (in 198 farms with ≥3000 birds) in the Netherlands were tested. A questionnaire about farm characteristics was also administered. Sampling was performed in July 2018-May 2019, three weeks before slaughter. Fourteen flocks (in 10 farms) were S. Infantis-positive, resulting in a 3.7 % flock-level and 5.1 % farm-level prevalence. Based on multi-locus sequence typing (MLST), all isolates belonged to sequence type 32. All but one isolate carried a pESI-like mega-plasmid. Core-genome MLST showed considerable heterogeneity among the isolates, even within the same farm, with a few small clusters detected. The typical pESI-borne multi-resistance pattern to aminoglycosides, sulphonamide and tetracycline (93 %), as well as trimethoprim (71 %), was found. Additionally, resistance to (fluoro)quinolones based on gyrA gene mutations was detected. S. Infantis was found more often in flocks using salinomycin as coccidiostat, where flock thinning was applied or litter quality was poor, whereas employing external cleaning companies, wheat in feed, and vaccination against infectious bronchitis, were protective. Suggestive evidence for vertical transmission from hatcheries was found. A heterogeneous (pESI-carrying) S. Infantis population has established itself in Dutch broiler flocks, calling for further monitoring of its spread and a comprehensive appraisal of control options.

Summary Objectives To determine the contributions of several animal and environmental sources of human campylobacteriosis and identify source-specific risk factors. Methods 1417 Campylobacter jejuni/coli isolates from the Netherlands in 2017–2019 were whole-genome sequenced, including isolates from human cases (n = 280), chickens/turkeys (n = 238), laying hens (n = 56), cattle (n = 158), veal calves (n = 49), sheep/goats (n = 111), pigs (n = 110), dogs/cats (n = 100), wild birds (n = 62), and surface water (n = 253). Questionnaire-based exposure data was collected. Source attribution was performed using core-genome multilocus sequence typing. Risk factors were determined on the attribution estimates. Results Cases were mostly attributed to chickens/turkeys (48.2%), dogs/cats (18.0%), cattle (12.1%), and surface water (8.5%). Of the associations identified, never consuming chicken, as well as frequent chicken consumption, and rarely washing hands after touching raw meat, were risk factors for chicken/turkey-attributable infections. Consuming unpasteurized milk or barbecued beef increased the risk for cattle-attributable infections. Risk factors for infections attributable to environmental sources were open water swimming, contact with dog faeces, and consuming non-chicken/turkey avian meat like game birds. Conclusions Poultry and cattle are the main livestock sources of campylobacteriosis, while pets and surface water are important non-livestock sources. Foodborne transmission is only partially consistent with the attributions, as frequency and alternative pathways of exposure are significant.

Salmonella spp. infection has shown to have oncogenic transformative effects and thereby increases the risk of certain cancers. For Campylobacter spp., similar effects have been demonstrated. Risk factor identification may allow for timely diagnosis and preventive treatment. To substantiate the oncogenic potential of Salmonella and Campylobacter spp., this study compared the incidence of extrahepatic biliary tract cancer (BTC) in patients with diagnosed Salmonella or Campylobacter spp. infection with BTC incidence in the Netherlands. National infectious diseases surveillance records of patients diagnosed with a laboratory-confirmed Salmonella or Campylobacter spp. infection during 1999&ndash, 2016 were linked to the Netherlands Cancer Registry. Incidence of BTC in Salmonella and Campylobacter spp. patients was compared to the incidence of BTC in the general population using Standardized Incidence Ratios (SIRs). In total, 16,252 patients were diagnosed with Salmonella spp. and 27,668 with Campylobacter spp. infection. Nine patients developed BTC at a median of 46 months (13&ndash, 67) after Salmonella spp. infection and seven at a median of 60 months (18&ndash, 138) after Campylobacter spp. infection. SIR of BTC in salmonellosis patients was 1.53 (95% CI 0.70&ndash, 2.91). In patients aged &lt, 60 years, the SIR was 1.74 (95% CI 0.36&ndash, 5.04). For campylobacteriosis patients, the SIR was 0.97 (95% CI 0.39&ndash, 2.00). Even though Salmonella or Campylobacter spp. infection was not significantly associated with increased BTC risk in this cohort, it remains extremely important to study potential risk factors for cancer to facilitate screening and ultimately improve prognosis of cancer patients.

In Venezuela, areas endemic for schistosomiasis are of low transmission, with low parasite loads. Immunological tests often lack specificity and cannot differentiate past from present infections. Molecular tests are an alternative, although validation studies in endemic areas are needed. The aim of this study was to determine the performance of parasitological, immunological and molecular tests for the diagnosis of Schistosoma mansoni infection in low-transmission settings. A cross-sectional study was carried out in a rural community located in a schistosomiasis-endemic area of Venezuela to determine the prevalence and diagnostic performance of the Kato-Katz (KK) technique, Circumoval Precipitin Test (COPT), ELISA based on soluble egg antigen (ELISA-SEA) with and without treatment with sodium metaperiodate (ELISA-SEA-SMP), and PCR for amplification of the 121 bp highly repeated sequence of Schistosoma mansoni in faeces, urine and serum samples. The highest prevalence rates were obtained with ELISA-SEA (38.7%), COPT (33.3%), ELISA-SEA-SMP (31.5%), PCR on faeces (21.6%), and KK (17.1%), whereas PCR-based prevalence in urine was 6.2% and no positivity was detected in serum samples. Results showed that ELISA-SEA is the best method for the diagnosis of both current and former infections and that PCR on faeces is the best method for detecting recent transmission. The use of different tests that complement one another also allowed for a better diagnosis of Schistosoma mansoni infection, revealing a relatively high prevalence (33.8%) of schistosomiasis in a community of low transmission.

Listeriosis is a foodborne illness characterized by a relatively low morbidity, but a large disease burden due to the severity of clinical manifestations and the high case fatality rate. Increased listeriosis notifications have been observed in Europe since the 2000s. However, the reasons for this increase are largely unknown, with the sources of sporadic human listerioris often remaining elusive. Here we inferred the relative contributions of several putative sources of Listeria monocytogenes strains from listerioris patients in Northern Italy (Piedmont and Lombardy regions), using two established source attribution models (i.e. 'Dutch' and 'STRUCTURE') in comparative fashion. We compared the Multi-Locus Sequence Typing and Multi-Virulence-Locus Sequence Typing profiles of strains collected from beef, dairy, fish, game, mixed foods, mixed meat, pork, and poultry. Overall, 634 L. monocytogenes isolates were collected from 2005 to 2016. In total, 40 clonal complexes and 51 virulence types were identified, with 36% of the isolates belonging to possible epidemic clones (i.e. genetically related strains from unrelated outbreaks). Source attribution analysis showed that 50% of human listerioris cases (95% Confidence Interval 44-55%) could be attributed to dairy products, followed by poultry and pork (15% each), and mixed foods (15%). Since the contamination of dairy, poultry and pork products are closely linked to primary production, expanding actions currently limited to ready-to-eat products to the reservoir level may help reducing the risk of cross-contamination at the consumer level.

Background Staphylococcus pseudintermedius is a common pathogen in dogs and methicillin resistance has emerged over recent decades. According to the current guidelines, S. pseudintermedius displaying oxacillin resistance should be reported as resistant to all β-lactams. Objectives To identify possible associations between β-lactam resistance levels and clonal complexes (CCs) and/or staphylococcal cassette chromosome mec (SCCmec) types in methicillin-resistant S. pseudintermedius (MRSP). Methods MICs of oxacillin, penicillin, ampicillin, amoxicillin/clavulanic acid and cefalotin were determined by broth microdilution for 86 clinical canine MRSP isolates from Denmark and the Netherlands. PCR and sequencing were used for SCCmec typing and MLST. Results Isolates belonged to CC71 (n = 36), CC258 (n = 33), CC45 (n = 11), CC68 (n = 1) and five singleton STs. SCCmecII-III was exclusively found in CC71 and SCCmecIV was significantly associated with CC258. SCCmecV and non-typeable SCCmec types occurred in 4 and 14 isolates, respectively. SCCmecIV was associated with lower MICs of oxacillin ( Conclusions SCCmec types were associated with different CCs and with either high- or low-level resistance to different β-lactams. The finding of amoxicillin/clavulanic acid (20%) and cefalotin (70%) in vitro susceptibility across all CCs might have clinical implications, since amoxicillin/clavulanic acid and first-generation cephalosporins are first-choice antibiotics for treatment of S. pseudintermedius infections. Pharmacokinetic/pharmacodynamic and clinical outcome studies are warranted to evaluate the in vivo efficacy of these β-lactams for treatment of MRSP infections.

Presence of extended-spectrum β-lactamase (ESBL)- and AmpC β-lactamase (pAmpC)-producing Escherichia coli (ESBL/pAmpC-EC) in humans and animals is alarming due to the associated risks of antibiotic therapy failure. ESBL/pAmpC-EC transmission between the human and animal compartments remains controversial. Using cefotaxime-supplemented (selective) media, we recently showed high sample prevalence of ESBL/pAmpC-EC in an integrated broiler chain [i.e. Parent Stock (PS), offspring broilers and their carcasses]. Here, we used a different approach. In parallel with the selective isolation, samples were processed on non-selective media. E. coli isolates were tested for ESBL/pAmpC-production and those found positive were genotyped. For carcasses, total E. coli were enumerated. This approach enabled us to estimate prevalence at the isolate level, which mirrors ESBL/pAmpC-EC colonisation levels. We showed that although present in many animals, ESBL/pAmpC-EC were overall subdominant to intestinal E. coli, indicating that high sample prevalence is not associated with high levels of resistance in individual hosts. This is a relevant aspect for risk assessments, especially regarding the immediate exposure of farm personnel. An exception was a particularly dominant B2/blaCMY-2 lineage in the gut of imported PS chicks. This predominance obscured presence of latent genotypes, however bias towards particular ESBL/pAmpC-EC genotypes from the selective method or underestimation by the non-selective approach did not occur. At the slaughterhouse, we showed a link between total E. coli and ESBL/pAmpC-EC on carcasses. Mitigation strategies for reducing consumers’ exposure should aim at suppressing ESBL/pAmpC-EC in the broiler gut as well as controlling critical points in the processing line.

The IncK plasmid group can be divided into two separate lineages named IncK1 and IncK2. IncK2 is found predominantly in poultry while IncK1 was reported in various mammals, including animals and humans. The physiological basis of this distinction is not known. In this manuscript we examined fitness cost of IncK1 and IncK2 plasmids at 37 and 42°C, which resembles mammalian and chicken body temperatures, respectively. We analyzed conjugation frequency, plasmid copy number and plasmid fitness cost in direct competition. Additionally, we measured levels of σ-32 in Escherichia coli carrying either wild type or conjugation-deficient IncK plasmids. The results show that IncK2 plasmids have a higher conjugation frequency and lower copy number at 42°C compared to IncK1. While the overall fitness cost to the host bacterium of IncK2 plasmids was higher than that of IncK1, it was not affected by the temperature while the fitness cost of IncK1 was shown to increase at 42°C compared to 37°C. These differences correlate with an increased expression of σ-32, a regulator of heat-shock protein expression, in E. coli with IncK2 compared to cells containing IncK1. This effect was not seen in cells containing conjugation deficient plasmids. Therefore, it is hypothesized that the assembly of the functional T4S may lead to these increased levels of σ–32. Increased activation of CpxR at 42°C may explain why IncK2 plasmids, and not IncK1, are predominantly found in chicken isolates.

Poultry are the main reservoir for thermophilic Campylobacter spp., which is the most common causative agent of human bacterial gastroenteritis. The epidemiology of Campylobacter in poultry, particularly in turkeys, is not completely understood. This study aimed at identifying potential sources and transmission routes of thermophilic Campylobacter spp. in commercial turkey farms. C. jejuni and C. coli isolates from breeders (n = 29, 20 C. jejuni and 9 C. coli) and their progeny (n = 51, 18 C. jejuni and 33 C. coli) reared in two different farms for three sequential production cycles were analysed by multilocus sequence typing (MLST). Strains (n = 88, 42 C. jejuni and 46 C. coli) isolated from environmental (i.e. anteroom and in-house overshoes), water (i.e. drinkers and water line), and pest (i.e. flies, Alphitobius diaperinus, and mice) sources were also examined. MLST of C. jejuni and C. coli isolates resulted in 13 and 12 different sequence types (STs) belonging to six and one previously-described clonal complexes (CCs), respectively. Three novel STs were identified. Genetic similarities were detected between isolates from fattening turkeys and the considered environmental, water, and pest sources, and with the breeders to a lesser extent. Source attribution analysis estimated that environmental and water sources accounted for most (∼75%) of fattening turkey isolates and were therefore identified as the most likely sources of flock colonization, followed by pests (∼20%) and breeders (∼5%). These sources may thus be targeted by control measures to mitigate the risk of Campylobacter colonization in commercial turkeys. RESEARCH HIGHLIGHTSHigh occurrence of C. jejuni and C. coli in commercial turkey flocks.High genetic diversity of C. jejuni and C. coli in commercial turkey flocks.Horizontal transmission responsible for Campylobacter colonization of commercial turkey flocks.Environmental and water sources involved in Campylobacter colonization of commercial turkey flocks.Strategies for prevention and control of Campylobacter colonization of commercial turkey flocks are needed. High occurrence of C. jejuni and C. coli in commercial turkey flocks. High genetic diversity of C. jejuni and C. coli in commercial turkey flocks. Horizontal transmission responsible for Campylobacter colonization of commercial turkey flocks. Environmental and water sources involved in Campylobacter colonization of commercial turkey flocks. Strategies for prevention and control of Campylobacter colonization of commercial turkey flocks are needed.

Highlights • CPV, CCoV and β-hemolytic E. coli are associated with (severe) acute diarrhoea in puppies. • Puppies from high-volume breeders are more likely to be infected with CPV, CCoV and Cystoisospora spp. • CPV and CCoV are more often detected in winter/spring. • Multiple enteric infections are common in puppies with and without diarrhoea. • Screening of CPV, CCoV and β-hemolytic E. coli is advised in diarrhoeic puppies., Laboratory confirmation of the causative agent(s) of diarrhoea in puppies may allow for appropriate treatment. The presence of potential pathogens however, does not prove a causal relationship with diarrhoea. The aim of this study was to identify specific enteropathogens in ≤12 month old puppies with and without acute diarrhoea and to assess their associations with clinical signs, putative risk factors and pathogen co-occurrence. Faecal samples from puppies with (n = 113) and without (n = 56) acute diarrhoea were collected and screened for Canine Parvovirus (CPV), Canine Coronavirus (CCoV), Salmonella spp., Campylobacter spp., Clostridium perfringens, Clostridium difficile, β-hemolytic Eschericha coli (hEC), Giardia spp., Toxocara spp., Cystoisospora spp., and Cyniclomyces guttulatus. One or more pathogens were detected in 86.5% of diarrhoeic puppies and in 77.8% of asymptomatic puppies. Significant positive associations were found between CPV and CCoV, CPV and Cystoisospora spp., Toxocara spp. and hEC, Giardia spp. and C. guttulatus. Only CPV and CCoV were significantly associated with diarrhoea, hEC with a subset of puppies that had diarrhoea and severe clinical signs. CPV was more prevalent in puppies under 3 months of age. Puppies from high-volume dog breeders were significantly at increased risk for CPV (OR 4.20), CCoV (OR 4.50) and Cystoisospora spp. (OR 3.60). CCoV occurred significantly more often in winter (OR 3.35), and CPV in winter (OR 3.78) and spring (OR 4.72) as compared to summer. We conclude that routine screening for CPV, CCoV and hEC is recommended in puppies with acute diarrhoea, especially if they are under 3 months of age and originate from high-volume dog breeders. Routine screening for other pathogens may lead to less conclusive results.

Antimicrobial use (AMU) in humans and animals facilitates the emergence of antimicrobial resistance (AMR). With increasing AMR being recognised as a major global threat for public health, responsible AMU is strongly advocated in both human and veterinary medicine. Knowledge on factors influencing antimicrobial prescribing behaviour of companion animal veterinarians is needed to promote responsible AMU in companion animals and to improve compliance with current legislation and guidelines. The present study aimed to quantitively investigate attitudes and perceptions of companion animal veterinarians towards AMU and AMR and to identify associations with demographic characteristics as possible explanatory variables. A self-administered questionnaire was developed based upon an earlier qualitative interview study, and 1608 potential participants (i.e. practising companion animal veterinarians) were invited. The questionnaire included questions addressing general descriptives of the respondents and questions with 6-point Likert scale statements, to assess attitudes towards AMU, AMR, factors influencing antimicrobial prescribing, and possible options to support responsible AMU. The response rate was 32% (22% when complete questionnaires considered). Categorical Principal Component Analysis (CATPCA) was conducted on 76 Likert scale questions. This resulted in a final model with 37 questions explaining 38.7% of the variance of the question scores, with three underlying dimensions ("attitudinal profiles"). Additionally, general descriptives were added to the CATPCA as possible explanatory variables. The first dimension, related to "social responsibility" was positively associated with veterinarians working in clinics dedicated to companion animals, with veterinarians working in a referral clinic, and with more experienced veterinarians. The second dimension was related to "scepticism", which was positively associated with being a male veterinarian and with more experienced veterinarians. The third dimension was related to "risk avoidance", especially regarding surgical procedures, and was negatively associated with veterinarians working in clinics in urban areas and with veterinarians working part-time. Antimicrobial prescribing behaviour was self-reported to be well considered, and respondents did not see economic drivers as important influencing factors. The unwillingness of owners and financial constraints were perceived as important barriers for performing further diagnostics. To improve AMU, a multifaceted approach, taking differences between companion animal veterinarians (e.g., in experience and gender) and differences in work situation (e.g., full-time versus part-time) into account, should be directed at companion animal veterinarians and owners. Moreover, a joint and comprehensive effort of several stakeholders, like veterinary nurses, guideline developers, pharmaceutical industry, and providers of diagnostics, is needed to optimise AMU in companion animals.

Background The epidemiology of carriage of extended-spectrum beta-lactamase-producing (ESBL-E) and carbapenemase-producing Enterobacteriaceae (CPE) in the general population is unknown. Aim In this observational study, the prevalence and risk factors for intestinal ESBL-E and CPE carriage in the Dutch general population were determined. ESBL-E were characterised. Methods From 2014 to 2016, ca 2,000 residents were invited monthly to complete a questionnaire and provide a faecal sample, which was tested for ESBL-E. The first 1,758 samples were also tested for CPE. Risk factors for ESBL-E carriage were identified by multivariable logistic regression analysis. ESBL-E isolates underwent whole genome sequencing. Results Of 47,957 individuals invited, 4,177 (8.7%) completed the questionnaire and provided a faecal sample. ESBL-E were detected in 186 (4.5%) individuals, resulting in an adjusted prevalence of 5.0% (95% confidence interval (CI):3.4–6.6%). Risk factors were: born outside the Netherlands (odds ratio (OR): 1.99; 95% CI: 1.16−4.54), eating in restaurants > 20 times/year (OR: 1.70; 95% CI: 1.04−2.76), antibiotic use bla CTX-M-15. Escherichia coli isolates belonged to 70 different sequence types (ST)s, of which ST131 (42/178 isolates; 23.6%) was most prevalent. Associations were observed between IncFIA plasmids and ST131 and bla CTX-M-27, and between IncI1 and ST88 and bla CTX-M-1. No CPE were detected. Conclusions The prevalence of ESBL-E carriage in the Netherlands’ community-dwelling population is 5.0%. Identified risk factors were mostly travelling (particularly to Asia and Africa) and kitchen hygiene. CPE were not detected.

This study aimed at estimating gastroenteritis (GE) incidence in all age groups of the Netherlands' general population, with special emphasis on the role of children in GE burden, and the associated costs. Monthly from November 2014 to November 2016, a random sample of 2000 residents in the Netherlands was invited to complete a questionnaire on household characteristics and health complaints. We calculated GE incidence rates standardized to the Dutch population and used multivariable logistic regression models to identify potential risk factors. We calculated the costs related to resources used within the healthcare sector, the resources used by patients and their families, and productivity losses (paid worktime) due to GE. The overall standardized incidence rate was 0.81 GE episodes/person-year, with the highest rate in children ≤4 years (1.96 episodes/person-year). GE was observed more often in households with children (≤17 years), especially if children attended out-of-home childcare services, and among individuals with non-native Dutch ethnic background. Less GE was observed among employed persons aged 25-64 years, compared with those unemployed, but the opposite was observed in persons ≥65 years. The average costs per GE episode was €191, resulting in €945 million annual total costs for GE in the Netherlands (€55 per inhabitant). The majority of costs (55%) were attributable to productivity losses of the ill or their caregivers. In conclusion, GE still poses a significant burden, particularly in preschool children and adults living in households with children. Similar to other industrialized countries, the major factor driving the costs due to GE was the loss of productivity. This study also provides up-to-date baseline GE incidence rates and associated societal costs to better contextualize the burden of the disease in support of policy making.

Background Climate change is expected to increase the chance of extreme rainfall events in the Northern Hemisphere and herewith, there is an increased chance of urban pluvial flooding. Urban pluvial flooding often consists of street flooding and/or flooding of combined sewerage systems, leading to contamination of the floodwater with several gastrointestinal and/or respiratory pathogens. An increase in flooding events therefore pose a health risk to those exposed to urban floodwater. We studied the association between exposure to pluvial floodwater and acute gastroenteritis (AGE) and acute respiratory infection (ARI). Methods We performed a retrospective, cross-sectional survey during the summer of 2015 in 60 locations in the Netherlands with reported flooding. Two weeks after the flooding, questionnaires were sent to households in these locations, collecting data on self-reported AGE and ARI and information on floodwater exposure in the previous 2 weeks. Multivariable generalized estimating equations (GEE) regression models, accounting for the clustered data structure, were used to identify risk factors for AGE and ARI. Results In total, 699 households with 1,656 participants (response rate 21%) returned the questionnaire. Contact with floodwater was significantly associated with AGE (aOR 4.2, 95%CI 2.1–8.4) and ARI (aOR 3.3, 95%CI 2.0–5.4). Risk factors for AGE were skin contact with floodwater (aOR 4.0, 95%CI 1.8-9.0), performing post-flooding cleaning operations (aOR 8.6, 95%CI 3.5-20.9) and cycling through floodwater (aOR 2.3, 95%CI 1.0-5.0). Skin contact with floodwater (aOR 3.6, 95%CI 1.9-6.9) and performing post-flooding cleaning operations (aOR 5.5, 95%CI 3.0-10.3) were identified as risk factors for ARI. Conclusions Results suggest an association between direct exposure to pluvial floodwater and AGE and ARI. As it is predicted that the frequency of pluvial flooding events will increase in the future, there is a need for flood-proof solutions in urban development and increased awareness among stakeholders and the public about the potential health risks. Future prospective studies are recommended to confirm our results. Electronic supplementary material The online version of this article (10.1186/s12879-019-3984-5) contains supplementary material, which is available to authorized users.

Extended-spectrum β-lactamase (ESBL)- and plasmid mediated AmpC-type cephalosporinase (pAmpC)-producing Escherichia coli (ESBL/pAmpC E. coli) in food-producing animals is a major public health concern. This study aimed at quantifying ESBL/pAmpC-E. coli occurrence and transfer in Italy's broiler production pyramid. Three production chains of an integrated broiler company were investigated. Cloacal swabs were taken from parent stock chickens and offspring broiler flocks in four fattening farms per chain. Carcasses from sampled broiler flocks were collected at slaughterhouse. Samples were processed on selective media, and E. coli colonies were screened for ESBL/pAmpC production. ESBL/pAmpC genes and E. coli phylogroups were determined by PCR and sequencing. Average pairwise overlap of ESBL/pAmpC E. coli gene and phylogroup occurrences between subsequent production stages was estimated using the proportional similarity index, modelling uncertainty in a Monte Carlo simulation setting. In total, 820 samples were processed, from which 513 ESBL/pAmpC E. coli isolates were obtained. We found a high prevalence (92.5%, 95% CI 72.1-98.3%) in day-old parent stock chicks, in which blaCMY-2 predominated; prevalence then dropped to 20% (12.9-29.6%) at laying phase. In fattening broilers, prevalence was 69.2% (53.6-81.3%) at the start of production, 54.2% (38.9-68.6%) at slaughter time, and 61.3% (48.1-72.9%) in carcasses. Significantly decreasing and increasing trends for respectively blaCMY-2 and blaCTX-M-1 gene occurrences were found across subsequent production stages. ESBL/pAmpC E. coli genetic background appeared complex and bla-gene/ phylogroup associations indicated clonal and horizontal transmission. Modelling revealed that the average transfer of ESBL/pAmpC E. coli genes between subsequent production stages was 47.7% (42.3-53.4%). We concluded that ESBL/pAmpC E. coli in the broiler production pyramid is prevalent, with substantial transfer between subsequent production levels.

Campylobacter jejuni and C. coli, the primary agents of human bacterial gastroenteritis worldwide, are widespread in surface water. Several animal sources contribute to surface water contamination with Campylobacter, but their relative contributions thus far remained unclear. Here, the prevalence, genotype diversity, and potential animal sources of C. jejuni and C. coli strains in surface water in the Netherlands were investigated. It was also assessed whether the contribution of the different animal sources varied according to surface water type (i.e. agricultural water, surface water at discharge points of wastewater treatment plants [WWTPs], and official recreational water), season, and local livestock (poultry, pig, ruminant) density. For each surface water type, 30 locations spread over six areas with either high or low density of poultry, ruminants, or pigs, were sampled once every season in 2018-2019. Campylobacter prevalence was highest in agricultural waters (77%), and in autumn and winter (74%), and lowest in recreational waters (46%) and in summer (54%). In total, 76 C. jejuni and 177 C. coli water isolates were whole-genome sequenced. Most C. coli water isolates (78.5%) belonged to hitherto unidentified clones when using the seven-locus sequence type (ST) scheme, while only 11.8% of the C. jejuni isolates had unidentified STs. The origin of these isolates, as defined by core-genome multi-locus sequence typing (cgMLST), was inferred by comparison with Campylobacter strain collections from meat-producing poultry, laying hens, adult cattle, veal calves, small ruminants, pigs, and wild birds. Water isolates were mainly attributed to wild birds (C. jejuni: 60.0%; C. coli: 93.7%) and meat-producing poultry (C. jejuni: 18.9%; C. coli: 5.6%). Wild bird contribution was high among isolates from recreational waters and WWTP discharge points, and in areas with low poultry (C. coli) or high ruminant (C. jejuni) densities. The contribution of meat-producing poultry was high in areas with high density of poultry, springtime, agricultural waters and WWTP discharge points. While wild birds and poultry were the main contributors to Campylobacter contamination in surface water, their contribution differed significantly by water type, season, and local poultry and ruminant densities.

Background: We aimed to estimate population-level exposure to Campylobacter and associated risk factors, using three approaches for serological data analysis. Methods: Nationwide, population-based serosurvey in the Netherlands (Feb 2006–Jun 2007). Anti-Campylobacter IgG, IgM and IgA were measured using ELISA, and analysed via: a) seroincidence estimation, using reference values of antibody peak levels and decay rates over-time after Campylobacter exposure; b) two normal distributions of true positives/negatives fitted to the IgG distribution to derive seroprevalence and individual probability of being positive/negative; and c) IgG levels. Risk factors were analysed using multiple linear regressions. Results: From 1559 respondents, seroincidence was estimated at 1.61 infections/person-year (95%CI:1.58–1.64) and seroprevalence at 68.1% (65.4–70.9). The three approaches identified similar risk factors, although seroincidence had higher power and results were interpretable as risk: seroincidence was higher in females [exp(b) = 1.07(1.04–1.11)], older ages [vs. 15–34 years; for < 5, 5–14, 35–54 and 55–70 years: 0.60(0.58–0.63), 0.74(0.71–0.78), 1.08(1.03–1.13) and 1.08(1.01–1.16), respectively], non-Dutch background [Moroccan/Turkish: 1.25(1.14–1.37); Caribbean: 1.14(1.03–1.25)], low socioeconomic status [1.05(1.01–1.10)], traveling outside Europe [1.05(1.01–1.09)], and eating undercooked meat [1.04(1.01–1.08)]. Conclusion: Campylobacter exposure is much higher than clinical infection rates, but risk factors are similar to those previously described.Seroincidence is a powerful measure to study Campylobacter epidemiology, and is preferred over other methods.

Correlates of protection (CoPs) against the highly contagious respiratory disease whooping cough, caused by Bordetella pertussis, remain elusive. Characterizing the antibody response to this pathogen is essential towards identifying potential CoPs. Here, we evaluate levels, avidity and functionality of B. pertussis-specific-antibodies from paired plasma samples derived from symptomatic and recovered pertussis patients, as well as controls. Natural infection is expected to induce protective immunity. IgG levels and avidity to nine B. pertussis antigens were determined using a novel multiplex panel. Furthermore, opsonophagocytosis of a B. pertussis clinical isolate by neutrophils was measured. Findings indicate that following infection, B. pertussis-specific antibody levels of (ex-) pertussis patients waned, while the avidity of antibodies directed against the majority of studied antigens increased. Opsonophagocytosis indices decreased upon recovery, but remained higher than controls. Random forest analysis of all the data revealed that 28% of the opsonophagocytosis index variances could be explained by filamentous hemagglutinin- followed by pertussis toxin-specific antibodies. We propose to further explore which other B. pertussis-specific antibodies can better predict opsonophagocytosis. Moreover, other B. pertussis-specific antibody functions as well as the possible integration of these functions in combination with other immune cell properties should be evaluated towards the identification of CoPs against pertussis.

Microbial subtyping is the most common approach for Salmonella source attribution. Typically, attributions are computed using frequency-matching models like the Dutch and Danish models based on phenotyping data (serotyping, phage-typing, and antimicrobial resistance profiling). Herewith, we critically review three major paradigms facing Salmonella source attribution today: (i) the use of genotyping data, particularly Multi-Locus Variable Number of Tandem Repeats Analysis (MLVA), which is replacing traditional Salmonella phenotyping beyond serotyping; (ii) the integration of case-control data into source attribution to improve risk factor identification/characterization; (iii) the investigation of non-food sources, as attributions tend to focus on foods of animal origin only. Population genetics models or simplified MLVA schemes may provide feasible options for source attribution, although there is a strong need to explore novel modelling options as we move towards whole-genome sequencing as the standard. Classical case-control studies are enhanced by incorporating source attribution results, as individuals acquiring salmonellosis from different sources have different associated risk factors. Thus, the more such analyses are performed the better Salmonella epidemiology will be understood. Reparametrizing current models allows for inclusion of sources like reptiles, the study of which improves our understanding of Salmonella epidemiology beyond food to tackle the pathogen in a more holistic way.

In January 2017, an increase in reported Salmonella enterica serotype Bovismorbificans cases in the Netherlands was observed since October 2016. We implemented a case–control study to identify the source, including all cases after December 2016. Adjusted odds ratios were calculated using logistic regression analysis. We traced back the distribution chain of suspected food items and sampled them for microbiological analysis. Human and food isolates were sequenced using whole genome sequencing (WGS). From October 2016 to March 2017, 54 S. Bovismorbificans cases were identified. Sequencing indicated that all were infected with identical strains. Twenty-four cases and 37 controls participated in the study. Cases were more likely to have consumed ham products than controls (aOR = 13; 95% CI: 2.0–77) and to have shopped at a supermarket chain (aOR = 7; 95% CI: 1.3–38). Trace-back investigations led to a Belgian meat processor: one retail ham sample originating from this processor tested positive for S. Bovismorbificans and matched the outbreak strain by WGS. All ham products related to the same batch were removed from the market to prevent further cases. This investigation illustrates the importance of laboratory surveillance for all Salmonella serotypes and the usefulness of WGS in an outbreak investigation.

BACKGROUND: Colon cancer constitutes one of the most frequent malignancies. Previous studies showed that Salmonella manipulates host cell signaling pathways and that Salmonella Typhimurium infection facilitates colon cancer development in genetically predisposed mice. This epidemiological study examined whether severe Salmonella infection, usually acquired from contaminated food, is associated with increased colon cancer risk in humans.METHODS AND FINDINGS: We performed a nationwide registry-based study to assess colon cancer risk after diagnosed Salmonella infection. National infectious disease surveillance records (1999-2015) for Dutch residents aged ≥20 years when diagnosed with salmonellosis (n = 14,264) were linked to the Netherlands Cancer Registry. Salmonella-infected patients were laboratory-confirmed under medical consultation after 1-2 weeks of illness. These datasets also contained information on Salmonella serovar and type of infection. Colon cancer risk (overall and per colon subsite) among patients with a diagnosed Salmonella infection was compared with expected colon cancer risk in the general population. Data from the nationwide registry of histo- and cytopathology (PALGA) and Statistics Netherlands (CBS) allowed assessing potential effects of age, gender, latency, socioeconomic status, genetic predisposition, inflammatory bowel disease (IBD), and tumor features. We found that compared to the general population, colon cancer risk was significantly increased (standardized incidence ratio [SIR] 1.54; 95%CI 1.09-2.10) among patients with Salmonella infection diagnosed CONCLUSIONS: Patients diagnosed with severe salmonellosis have an increased risk of developing cancer in the ascending/transverse parts of the colon. This risk concerns particularly S. Enteritidis infection, suggesting a contribution of this major foodborne pathogen to colon cancer development.

Urban canal swimming events are popular in the Netherlands. In 2015, two city canal swimming events took place, in Utrecht (Utrecht Singel Swim, USS) and in Amsterdam (Amsterdam City Swim, ACS). This prospective study characterizes the health risks associated with swimming in urban waters. Online questionnaires were sent to 160 (USS) and 2,692 (ACS) participants, with relatives of participants who did not swim completing the questionnaire as a control. Swimming water specimens and stool specimens of diarrheic participants in the ACS group were analysed. A total of 49% of USS and 51% of ACS swimmers returned their questionnaires. Nine percent of USS swimmers and 4% of non-swimmers reported gastrointestinal complaints (aRR 2.1; 95% CI: 0.3-16), while a total of 31% of ACS swimmers and 5% of non-swimmers reported gastrointestinal complaints (aRR 6.3; 95% CI: 4.1-9.5). AGI risk among ACS participants was directly related to increasing number of mouthfuls of water swallowed. Various norovirus genotypes were detected in five out of seven stool specimens taken from ACS participants and in all three tested ACS water samples. We conclude that the AGI risk among open-water swimmers in urban areas depends on the circumstances around the event. The epidemiological curve, the statistical association between swimming and AGI, and the microbiological evidence for norovirus in stool and water specimens suggest that AGI outbreak after the ACS event was due to water contamination by multiple norovirus strains, which is possibly linked to sewage overflow due to prior heavy rainfall. There is need for more targeted preventive measurements and recommendations for organizers, municipal authorities and participants to prevent this reoccurring in the future.

Background To reduce environmental contamination with Toxocara canis eggs, the current general advice is to deworm all dogs older than six months on average four times a year. However, only a small proportion of non-juvenile household dogs actually shed T. canis eggs, and some dogs shed eggs more frequently than others. The identification of these frequent shedders and the associated risk factors is an important cornerstone for constructing evidence-based deworming regimens. The purpose of this study is to identify risk factors associated with recurrence of periods of shedding Toxocara eggs in a cohort of household dogs older than six months. Methods We performed a prospective study (July 2011 to October 2014) on shedding Toxocara eggs in a cohort of 938 household dogs older than six months from all over the Netherlands. The median follow-up time was 14 months. Monthly, owners sent faecal samples of their dogs for Toxocara testing and completed a questionnaire. Dogs were dewormed only after diagnosis of a patent infection (PI). Survival analysis was used to assess factors influencing the time to first diagnosed PIs (FPI) and the time to recurrent PIs (RPI). Results The overall prevalence of PIs was 4.5 %, resulting in an estimated average incidence of 0.54 PIs/dog/year. No PI was diagnosed in 67.9 % of the dogs, 17.5 % of the dogs went through only one PI and 14.6 % had > 1 PI. Prevalence of PIs always peaked during wintertime. Increased hazards for first diagnosed PIs were associated with coprophagy, geophagy, walking off-leash for ≥ 80 % of walking time, reported worms in the faeces, feeding a commercial diet and suffering from urologic or respiratory conditions. Median time to reinfection was nine months. Factors associated with increased hazards for recurrent PIs were taking corticosteroids, changing dog’s main purpose, and proxies for veterinary care-seeking behaviours. Conclusions We concluded that targeted anthelmintic treatments in household dogs may be feasible as PIs tend to (re)occur in specific periods and in groups of dogs at high risk. Moreover, recurrent PIs appear to be influenced more by factors related to impaired immunity than environmental exposure to Toxocara eggs. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1816-7) contains supplementary material, which is available to authorized users.

Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water contamination with Campylobacter is largely unknown. In the Netherlands, the massive poultry culling to control the 2003 avian influenza epidemic coincided with a 44-50% reduction in human campylobacteriosis cases in the culling areas, suggesting substantial environment-mediated spread of poultry-borne Campylobacter. We inferred the origin of surface water Campylobacter jejuni and Campylobacter coli strains in Luxembourg and the Netherlands, as defined by multilocus sequence typing, by comparison to strains from poultry, pigs, ruminants, and wild birds, using the asymmetric island model for source attribution. Most Luxembourgish water strains were attributed to wild birds (61.0%), followed by poultry (18.8%), ruminants (15.9%), and pigs (4.3%); whereas the Dutch water strains were mainly attributed to poultry (51.7%), wild birds (37.3%), ruminants (9.8%), and pigs (1.2%). Attributions varied over seasons and surface water types, and geographical variation in the relative contribution of poultry correlated with the magnitude of poultry production at either the national or provincial level, suggesting that environmental dissemination of Campylobacter from poultry farms and slaughterhouses can be substantial in poultry-rich regions.

BACKGROUND: The purpose of this study was to determine the prevalence and risk factors for colonization with extended-spectrum cephalosporin-resistant (ESC-R) Escherichia coli in daycare center (DCC)-attending children. METHODS: This is a prospective cohort study including 44 DCCs in the Netherlands, combining DCC characteristics and monthly collected stool samples from their attendees, and was performed in 2010-2012. During a 22-month study period, 852 stool samples were collected and screened for ESC-R E coli. Risk factors were studied using logistic regression analysis. RESULTS: In DCC-attending children (

Campylobacteriosis is a frequently diagnosed disease in humans. Most infections are considered food-borne and are caused by Campylobacter jejuni and C. coli. The animal reservoirs of these Campylobacter, and the sources and routes of transmission, are described and discussed. Most warm-blooded animals can be colonized by Campylobacter, but avians, and in particular poultry, are preferred hosts. Much of the world’s poultry production is colonized by Campylobacter. Source attribution studies estimate that 20–40 % of cases are attributed to the handling and consumption of chicken meat, while up to 80 % of cases are due to Campylobacter found in the chicken reservoir. The difference suggests that routes other than through the food chain, i.e. environmental contamination, are important. Thus the most effective interventions would be targeted to primary production. To date, only improved biosecurity is available. If effectively implemented strict biosecurity can reduce the number of Campylobacter-positive flocks, but implementation to this level has proved difficult for the poultry industry. Available interventions in chicken processing plants can substantially reduce Campylobacter numbers on carcasses and consequently reduce the risk to humans. Public health strategies therefore utilize control programs, which aim at reducing the level of Campylobacter by measures along the food chain. It is now recognized that commercially acceptable complementary interventions for primary production, such as vaccines, bacteriophages, feed additives, are urgently needed. Once Campylobacter in poultry is controlled then other minor sources of Campylobacter including contaminated drinking water, direct contact with (pet) animals, and other food items (e.g. red meat and milk), can be addressed.