A National audit of the care of patients with acute kidney injury in England and Wales in 2019 and the association with patient outcomes.

BACKGROUND: Acute kidney injury (AKI) is a common complication of hospitalisations. This national audit assessed the care received by patients with AKI in hospital Trusts in England and Wales. METHODS: Twenty four hospital Trusts across England and Wales took part. Patients with AKI stage2/3 were identified using the UK Renal Registry AKI master patient index. Data was returned through a secure portal with linkage to hospital episode statistic mortality and hospitalisation data. Completion rates of AKI care standards and regional variations in care were established. RESULTS: 989 AKI episodes were included in the analyses. In-hospital 30-day mortality was 31-33.1% (AKI 2/3). Standard AKI interventions were completed in >80% of episodes. Significant inter-hospital variation remained in attainment of AKI care standards after adjustment for age and sex. Recording of urinalysis (41.9%) and timely imaging (37.2%) were low. Information on discharge summaries relating to medication changes/re-commencement and follow-up blood tests associated with reduced mortality. No quality indicators relating to clinical management associated with mortality. Better communication on discharge summaries associated with reduced mortality. CONCLUSIONS: Outcomes for patients with AKI in hospital remain poor. Regional variation in care exists. Work is needed to assess whether improving and standardising care improves patient outcomes.

The role of ultrasound in the investigation of neonatal jaundice.

Infants with persistent conjugated jaundice, lasting longer than 14 days of age in a term infant and 21 days in a preterm infant, should be referred to a national paediatric liver unit for investigation of the cause. This paper reviews the ultrasound findings in such cases with a particular emphasis upon the diagnosis of biliary atresia.

Phylogenetic Analysis of Mycobacterium tuberculosis Strains in Wales by Use of Core Genome Multilocus Sequence Typing To Analyze Whole-Genome Sequencing Data.

An inability to standardize the bioinformatic data produced by whole-genome sequencing (WGS) has been a barrier to its widespread use in tuberculosis phylogenetics. The aim of this study was to carry out a phylogenetic analysis of tuberculosis in Wales, United Kingdom, using Ridom SeqSphere software for core genome multilocus sequence typing (cgMLST) analysis of whole-genome sequencing data. The phylogenetics of tuberculosis in Wales have not previously been studied. Sixty-six Mycobacterium tuberculosis isolates (including 42 outbreak-associated isolates) from south Wales were sequenced using an Illumina platform. Isolates were assigned to principal genetic groups, single nucleotide polymorphism (SNP) cluster groups, lineages, and sublineages using SNP-calling protocols. WGS data were submitted to the Ridom SeqSphere software for cgMLST analysis and analyzed alongside 179 previously lineage-defined isolates. The data set was dominated by the Euro-American lineage, with the sublineage composition being dominated by T, X, and Haarlem family strains. The cgMLST analysis successfully assigned 58 isolates to major lineages, and the results were consistent with those obtained by traditional SNP mapping methods. In addition, the cgMLST scheme was used to resolve an outbreak of tuberculosis occurring in the region. This study supports the use of a cgMLST method for standardized phylogenetic assignment of tuberculosis isolates and for outbreak resolution and provides the first insight into Welsh tuberculosis phylogenetics, identifying the presence of the Haarlem sublineage commonly associated with virulent traits.

Reliably colonising broiler chickens with Campylobacter spp. using a litter-based method.

1. Chicken-associated Campylobacter spp. are the cause of most food poisoning cases in Europe. In order to study the host-pathogen interactions, a reliable and reproducible method of colonising chickens with the bacteria is required. 2. This study aimed to identify a more appropriate and less invasive method of colonisation (cf. gavaging) by seeding bedding material (litter) that commercial chickens are kept on with a mixture of Campylobacter spp., broth and faeces. 3. The first phase of the study tested the longevity of Campylobacter spp. recovery in seeded litter over 24 h: significantly more Campylobacter spp. was recovered at 0 or 3 h post-seeding than at 6 and 24 h post-seeding, indicating that the pathogen can survive to detectable levels for at least 3 h in this environment. 4. In the second phase, three groups of 10 broiler chickens (negative for Campylobacter spp. prior to exposure) were exposed at 21 days of age to one of three different Campylobacter jejuni and C. coli mixes (A, B, C), using the method above. At 28 days of age, birds were euthanised by overdose of barbiturate or cervical dislocation, and livers and caeca removed for Campylobacter spp. assessment. 5. All liver and 28/30 caeca samples tested positive for Campylobacter spp., with mix A and C giving higher counts in the caeca than mix B. The method of euthanasia did not affect Campylobacter spp. counts. 6. In conclusion, a successful method for reliably colonising broiler chickens with Campylobacter spp. has been developed which negates the need for gavaging and is more representative of how contamination occurs in the field.

Climatic factors and prevalence of Campylobacter in commercial broiler flocks in Thailand.

Campylobacter are bacteria associated with human foodborne disease worldwide. Poultry and poultry products are generally considered as a main source of these organisms. Compared to temperate zones, baseline information on Campylobacter in tropical regions is limited. Thus, the objectives of the present study were 1) to determine the prevalence of Campylobacter in Thai broiler flocks and 2) to investigate the association between climatic factors (i.e., rainfall, ambient temperature, and relative humidity) and Campylobacter colonization status of broiler flocks in Thailand. A total of 442 commercial broiler flocks reared in the central and northeastern regions of Thailand during 2012 to 2014 were investigated. Campylobacter positive status was identified in 252 examined flocks (57.01%; 95% CI 52.39 to 61.63%). Prevalence of Campylobacter in the northeastern region (54.46%; 95% CI 44.76 to 63.83%) was slightly lower than that of the central region (57.77%; 95% CI 52.47 to 62.90%). More than 65% of Campylobacter positive flocks in the central and northeastern regions had within-flock prevalence higher than 75%. Generalized estimating equations (GEE) revealed that the increased rainfall and relative humidity were associated with the increase of Campylobacter colonization in broiler flocks (P ≤ 0.05), while no relationship between ambient temperature and Campylobacter colonization status was identified.

A role for flies (Diptera) in the transmission of Campylobacter to broilers?

Campylobacter is the leading cause of bacterial diarrhoeal disease worldwide, with raw and undercooked poultry meat and products the primary source of infection. Colonization of broiler chicken flocks with Campylobacter has proved difficult to prevent, even with high levels of biosecurity. Dipteran flies are proven carriers of Campylobacter and their ingress into broiler houses may contribute to its transmission to broiler chickens. However, this has not been investigated in the UK. Campylobacter was cultured from 2195 flies collected from four UK broiler farms. Of flies cultured individually, 0·22% [2/902, 95% confidence interval (CI) 0-0·53] were positive by culture for Campylobacter spp. Additionally, 1293 flies were grouped by family and cultured in 127 batches: 4/127 (3·15%, 95% CI 0·11-6·19) from three broiler farms were positive for Campylobacter. Multilocus sequence typing of isolates demonstrated that the flies were carrying broiler-associated sequence types, responsible for human enteric illness. Malaise traps were used to survey the dipteran species diversity on study farms and also revealed up to 612 flies present around broiler-house ventilation inlets over a 2-h period. Therefore, despite the low prevalence of Campylobacter cultured from flies, the risk of transmission by this route may be high, particularly during summer when fly populations are greatest.

The contribution of systemic Escherichia coli infection to the early mortalities of commercial broiler chickens.

Avian pathogenic Escherichia coli (APEC) are a substantial burden to the global poultry industry. APEC cause a syndromic poultry infection known as colibacillosis, which has been previously associated with broiler chickens over 2 weeks old. We recently reported that the intestinal tract of 1-day-old broilers harbours a rich reservoir of potentially pathogenic E. coli. Prior infections of the reproductive tract of breeders, egg hygiene and transportation all contribute to early colonization of the neonatal gut. Up to one-half of all flock deaths occur in the first week of production, but few data are available describing the contribution of E. coli. In the present study, all dead birds collected on the first daily welfare walk 48 and 72 h after chick placement underwent post-mortem examination. Diseased tissues were selectively cultured for E. coli and isolates subsequently virulotyped using 10 APEC virulence-associated genes (VAGs): astA, iss, irp2, iucD, papC, tsh, vat, cvi, sitA and ibeA. Approximately 70% of birds displayed signs of colibacillosis. Thirty distinct virulence profiles were identified among 157 E. coli. Isolates carried between zero and seven VAGs; ∼ 30% of E. coli isolates carried five to seven VAGs, with 12.7% sharing the same VAG profile (astA, iss, irp2, iucD, tsh, cvi and sitA). Overall, this study demonstrates the significant contribution of E. coli infections to early broiler mortalities. The identification of a diverse E. coli population is unsurprising based on our previous findings. This work emphasizes the need for an effective vaccination programme and provides preliminary data for vaccine production.

Infection of chickens with antimicrobial-resistant Salmonella enterica Typhimurium DT193 and monophasic Salmonella Typhimurium-like variants: an emerging risk to the poultry industry?

Antimicrobial-resistant Salmonella enterica poses a particular risk to public health, and in particular isolates belonging to clonal lineages such as Salmonella Typhimurium DT104 cause epidemics across species including poultry. In recent years, antimicrobial-resistant S. Typhimurium DT193 and specifically the monophasic S. Typhimurium-like variants of this phage type, serotypes 4,12:i:- and 4,5,12:i:-, have become an increasing risk to public health in Europe and the USA and now account for nearly one-half of human S. Typhimurium infections in the UK. Unlike S. Typhimurium that possesses two forms of flagella which can vary between phase 1 and phase 2 during infection, monophasic variants possess only phase 1 flagella. These monophasic antimicrobial-resistant variants have become a major problem in pig production but human cases have also been associated with poultry consumption and have been found in UK flocks through surveillance schemes since 2010. In this study we determined the ability of antimicrobial-resistant DT193 serotype 4,12:i:- and 4,5,12:i:- isolates from pigs to infect chickens. All isolates were found to colonize the caeca and liver. All but one isolate of serotype 4,5,12:i:- also infected the spleen. Levels of infection and pathology were comparable with those found with the virulent S. Typhimurium isolate 4/74. These findings indicate that both S. Typhimurium DT193 and monophasic variants of this phage type usually associated with pigs are capable of colonizing the chicken. This shows that both S. Typhimurium DT193 and monophasic variants represent a significant and potential emerging threat to poultry production from "spill-over" of these isolates from the pig industry or other sources.

Campylobacter infection has different outcomes in fast- and slow-growing broiler chickens.

Campylobacter spp. are frequently carried by poultry, but they are not believed to cause significant disease in these animals. Modern poultry breeds have been selected to grow rapidly under intensive conditions, but recently, consumers have moved toward purchasing birds produced in higher welfare, free-range or organic systems. Birds reared in these systems tend to be a slower growing breed and are fed a different diet. Birds reared in such systems are stocked at a lower density compared with the standard conventional broilers, and they have access to environmental enrichment, such as perches. In previous research, these slower growing birds have been shown to have different levels of Campylobacter carriage in commercial rearing conditions, but the reasons for, and effect of, these different levels are unknown; is it the bird breed, diet, or environmental conditions? In this study, experimental flocks of fast- and slow-growing breeds of broiler chickens were reared to a standard commercial slaughter weight, with their weight gain being measured during the growing period. At 21 days, birds were either infected with Campylobacter jejuni or given a placebo as control. Cohorts of birds were euthanatized at various intervals, and samples were taken for examination for Campylobacter. The fast-growing birds gained weight more rapidly than the slow-growing birds. By 2 days postinfection (dpi), C. jejuni was detected in the caeca and by enrichment from the liver and spleen samples from both breeds of birds. Low-level colonization persisted in the spleen and liver samples but was undetectable by 28 dpi. Fast- and slow-growing birds did not show detectably different levels of Campylobacter carriage. Infection with C. jejuni affected the incidence of hock marks and pododermatitis in both breeds of birds, but the differences were greater with the fast-growing breed compared with the uninfected control birds. In addition, the incidence of pododermatitis was significantly higher in Campylobacter-positive fast-growing birds than in their slower-growing counterparts. The results show that infection with Campylobacter can have an indirect welfare effect on birds via increased incidence of hock marks and pododermatitis.

Enrichment culture can bias the isolation of Campylobacter subtypes.

Enrichment culture is often used to isolate Campylobacter. This study compared isolation of Campylobacter spp. from 119 broiler chicken environments from two farms, using Preston and modified Exeter (mExeter) and modified Bolton (mBolton) enrichments. mExeter was significantly more effective in isolating Campylobacter spp. from the environmental samples compared to Preston (P<0.001) and mBolton (P<0.04) broths but there was no significant difference between the latter two methods (P>0.05). Enrichment broth type did not affect isolation from chicken faecal or soil and litter samples. C. jejuni was isolated from significantly more environmental samples using mExeter broth compared to Preston (P<0.01) and mBolton (P<0.003) broths; there was no difference between the latter two methods or between all methods for detection of C. coli (P>0.05). Only C. coli was isolated from the soil and litter samples and although both C. jejuni and C. coli were recovered from the faecal samples there was no effect of using different enrichment broths. The majority of samples where the same species had been isolated yielded the same or closely related genotypes as defined by pulsed-field gel electrophoresis. Isolates recovered using Preston and mBolton broths were less genetically diverse than those from mExeter broth. We conclude that the enrichment method used affects both the number and species of Campylobacter isolated from naturally contaminated samples.

Porcine intestinal epithelial responses to Campylobacter infection.

Campylobacter infection is estimated to cause diarrhoea in 1% of the population of developed countries every year, but our understanding of this disease has been hampered by a lack of a suitable animal model. Colostrum-deprived piglets have been suggested as models since they produce similar clinical signs to humans when infected but little information currently exists regarding the response of this species to Campylobacter at cellular and molecular level. This study shows that intestinal epithelial cells from both species respond in a similar manner to Campylobacter infection regarding invasion, induction of innate immune response and effect on barrier function.

SulA-induced filamentation in Salmonella enterica serovar Typhimurium: effects on SPI-1 expression and epithelial infection.

AIMS: Salmonella enterica serovar Typhimurium is capable of adopting a filamentous phenotype in response to damage. How this adaptive response affects bacterial virulence is unclear. We have examined the hypothesis that filamentation affects the ability of Salmonella to infect host cells. METHODS AND RESULTS: Expression of the cell division inhibitor SulA in Salm. Typhimurium SL1344 from an arabinose-inducible plasmid resulted in filamentation. We examined expression of the type 3 secretion system (T3SS) encoded by Salmonella pathogenicity island 1 (SPI-1) using SL1344 expressing a chromosomal PprgH-gfp reporter. Single cell analysis of SulA-induced SL1344 PprgH-gfp revealed a relationship between increasing cell length and decreasing propensity for prgH expression, but there was no evidence of a significant change in prgH expression evident at the whole population level. Filamentous Salm. Typhimurium were capable of initiating membrane ruffling on MDCK epithelial cells, but only nonfilamentous bacteria (< 6 µm) invade. CONCLUSIONS: Induction of SulA expression in Salmonella inhibits septation. Increasing filament length is associated with down-regulation of SPI-1 gene expression, but a significant proportion of filaments retain the ability to produce SPI-1 T3SS and induce membrane ruffles on epithelia. Despite an active SPI-1 T3SS, filamentous Salmonella are unable to invade epithelial cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Our findings that filamentous Salmonella can express an invasive phenotype but fail to invade cells suggest that their presence in food does not constitute an immediate risk of infection until septation occurs. The described SulA expression model provides a convenient model for studying the impact of filamentation in the absence of additional stresses.

Influence of season and geography on Campylobacter jejuni and C. coli subtypes in housed broiler flocks reared in Great Britain.

Geographical and seasonal variation in the incidence and prevalence of Campylobacter jejuni and C. coli in housed broiler flocks reared in Great Britain in 2004 to 2006 was investigated in this study. Ceca (30) from 797 flocks, not subject to prior partial depopulation and reared on 211 farms, were examined individually for the presence of Campylobacter spp. The best-fitting climatic factors explained approximately 46% of the prevalence of Campylobacter-colonized flocks at slaughter and consisted of a combination of temperature at slaughter, number of sunshine hours in placement month, and millimeters of rainfall in placement month. Positive flocks were more likely to be slaughtered between June and November than during the rest of the year and to be reared in northern Great Britain than in central or southern Great Britain. C. jejuni was identified in approximately 90% of flocks, and C. coli was present in 10% of flocks. The most common clonal complexes identified in 226 isolates typed by multilocus sequence typing (MLST) were ST-45, ST-21, ST-574, ST-443, and ST-828. Flocks slaughtered at the same time were more likely to have similar complexes, and ST-45 had a seasonal pattern, with the highest prevalence in June, and was also more likely to be present in flocks reared in northern Great Britain.

Sub-clinical infection with Salmonella in chickens differentially affects behaviour and welfare in three inbred strains.

1. Much evidence exists detailing how animals respond to pathogen challenge, yet information explaining how the various behavioural, immunological, and physiological systems in chickens interplay during such challenges remains limited. 2. To gain an understanding of this interplay while controlling for genetic variation, the current study collected a variety of behavioural, physiological and immunological measures from three inbred lines (P, O and N) of laying hens before and after a sub-clinical infection with Salmonella enterica Typhimurium at 56 d of age. For comparison, an equal number of control birds were inoculated with a Salmonella-free broth. To identify an underlying profile, which might result in reduced susceptibility to infection, data were also collected in the pre-infection period. Post-infection blood and faeces were collected at 1-d post infection (dpi) and faeces again at 8 dpi. Animals were killed 15 d after infection and faeces, caecal contents, and spleen were examined for the presence of Salmonella. 3. Statistical analysis was performed to identify pre- and post-infection differences between genetic lines, changes in bird behavioural patterns between the two periods, and associations between a positive test for Salmonella and the various response measures. 4. Tissues from Line P birds were more often negative for Salmonella than those from birds of other lines, though this was inconsistent and tissue-dependent. The P line was also characterised by relatively greater serum concentrations of immunoglobulins at 1 dpi and α(1)-acid glycoprotein at 15 dpi. In addition, P line birds were more timid and their growth was reduced during the pre-infection period suggesting the possibility of a profile with reduced susceptibility to the bacterial challenge. 5. The current work has identified correlations between attributes of chicken strains and improved clearance. Future work using hypothesis-based testing will be required to determine whether the identified correlations are causally related.

Risk factors for Campylobacter colonisation during rearing of broiler flocks in Great Britain.

We investigated the associations between Campylobacter colonisation and management practices and farm characteristics in 603 housed broiler batches originating from 137 farms in Great Britain. All study batches were the initial batch slaughtered from the selected house on enrolled farms. Between 1 and 15 batches were sampled from each farm throughout the study. A total of 34.2% of the batches was Campylobacter positive and multivariable multilevel logistic regression revealed that the risk of Campylobacter colonisation was highest in July (OR=3.4, CI95%:1.8; 6.4), August (OR=3.4, CI95%:1.9; 6.2) and September (OR=3.7, CI95%:1.9; 7.1). Cattle on or adjacent to the farm increased the risk (OR=1.7, CI95%:1.1; 2.7), whereas chlorinated drinking water reduced it (OR=0.5, CI95%:0.2; 0.9). If the first removed batch from the previous flock in the house had been Campylobacter positive, the first batch of the following flock was also more likely to be colonised (OR=3.2, CI95%:2.1; 4.9). This association was more likely due to a persistent risk practice or source of Campylobacter on the farm than a direct carry-over from previous flock.

Rising rates of obstetric interventions: exploring the determinants of induction of labour.

BACKGROUND: Rising rates of obstetric interventions in the UK are a concern for health-care providers and the public. Our aims were to identify the socio-demographic and clinical factors (case mix) predictive of one of the most common obstetric interventions, induction of labour (IOL), and quantify the extent to which observed rates can be explained by case mix factors. METHODS: We conducted a comparative analysis of induced and spontaneous labours, using contemporary clinical data from the Aberdeen Maternity and Neonatal Databank. Cases complicated by antenatal intrauterine death or a previous or planned caesarean section were excluded. In total, 17,736 cases were included in the analysis. RESULTS: In 5727 (32.3%) cases labour was induced and in 12,009 (67.7%) cases it was spontaneous. Multivariate logistic regression modelling was used. In total, 18 case mix factors were predictive of IOL. Among these were well-recognized clinical indications for IOL such as pre-labour rupture of membranes (OR 3.29, 95% CI 2.90, 3.73) and prolonged pregnancy (OR 4.15, 95% CI 3.82, 4.50) and previously unreported case mix factors (residing an intermediate distance and travel time from hospital) (OR 1.27, 95% CI 1.18, 1.37; BMI >35 OR 1.37, 95% CI 1.14, 1.65). Case mix explained 71.5% of the observed rate of IOL. CONCLUSIONS: More than one-quarter of the rate of IOL remains unexplained by case mix factors. This may be explained by women's preferences for care and clinicians' practice.

Campylobacter in housed broiler chickens: a longitudinal study of risk factors.

Infections by Campylobacter spp. are a major cause of gastrointestinal disease in the United Kingdom. Most cases are associated with the consumption of chicken that has become contaminated during production. We investigated the epidemiology of Campylobacter spp. in chickens in a 3-year longitudinal study of flocks reared on 30 farms in the United Kingdom. We used Generalized Linear Mixed Effect Models (GLMM) to investigate putative risk factors associated with incidence and prevalence of flock infection arising from farm and flock management and local environmental conditions during rearing. We used survival analysis to investigate infection events and associated risk factors over the course of the study using two marginal models - the independent increment approach, which assumed that individual infection events were independent; and a conditional approach, which assumed that events were conditional on those preceding. Models of flock prevalence were highly overdispersed suggesting that infection within flocks was aggregated. The key predictors of flock infection identified from the GLMM analyses were mean temperature and mean rainfall in the month of slaughter and also the presence of natural ventilation. Mean temperature in the month of slaughter was also a significant predictor of flock infection, although the analyses suggested that the risk in flocks increased in a unimodal way in relation to temperature, peaking at 12 degrees C. The extent of pad burn was also identified as a predictor in these analyses. We conclude that predicting prevalence within flocks with linear modelling approaches is likely to be difficult, but that it may be possible to predict when flocks are at risk of Campylobacter infection. This is a key first step in managing disease and reducing the risks posed to the human food chain.

Demographic determinants for Campylobacter infection in England and Wales: implications for future epidemiological studies.

Despite a significant public health burden the epidemiology of human Campylobacter infection remains blurred. The identification of demographic determinants for Campylobacter infection is therefore essential for identifying potential areas for intervention. Demographic data from an active, population-based sentinel surveillance system for Campylobacter infection (from 2000 until 2003, n=15 907) were compared with appropriate denominator data from the 2001 United Kingdom Census. Incidence was higher in males from birth until the late teens and in females from 20 to 36 years. Age- and gender-specific differences in Campylobacter incidence were observed in different ethnic and socioeconomic groups and hence are all major drivers for Campylobacter infection. Epidemiological studies on Campylobacter infection need to take these factors into consideration during design and analysis. The collation of detailed epidemiological data and its comparison with appropriate denominator data provides a valuable epidemiological tool for studying infection.

Persistence of Campylobacter species, strain types, antibiotic resistance and mechanisms of tetracycline resistance in poultry flocks treated with chlortetracycline.

OBJECTIVES: The aim of this study was to investigate the persistence of Campylobacter species, strain types, antibiotic resistance and mechanisms of tetracycline resistance in poultry flocks treated with chlortetracycline. METHODS: Three commercially reared broiler flocks, naturally colonized with Campylobacter, were treated with chlortetracycline under experimental conditions. The numbers of Campylobacter isolated, and the species, flaA short variable region allele, and antimicrobial resistance of isolates were determined. RESULTS: For two of three flocks, tetracycline-resistant strains predominated prior to chlortetracycline exposure. Presence of the antibiotic had no discernible effect on the numbers or types of Campylobacter and the tetracycline-resistant strains persisted in numbers similar to those observed before treatment. With all flocks, some faecal samples were obtained that contained no Campylobacter, irrespective of exposure to chlortetracycline; this was more common as the birds grew older. For the third flock, tetracycline-resistant Campylobacter were in the minority of samples before and during exposure to chlortetracycline, but at sampling times after this, no resistant strains were found in the treated (or untreated) birds, irrespective of exposure to the antibiotic. All tetracycline-resistant isolates (MICs 16 to >128 mg/L) contained tet(O) and, for some isolates, this was transferable to Campylobacter jejuni 81116. The efflux pump inhibitor PAbetaN reduced the MICs of tetracycline for these isolates by 4-fold, suggesting that an intact efflux pump, presumably CmeABC, is required for high-level tetracycline resistance. CONCLUSIONS: Our data indicate that chlortetracycline treatment does not eradicate tetracycline-resistant Campylobacter spp. from poultry. However, if a low number of resistant isolates are present, then the antibiotic pressure appears insufficient to select such strains as the dominant population.

Effects of repeated cycles of acid challenge and growth on the phenotype and virulence of Salmonella enterica.

AIMS: The aim of the study was to investigate how stresses like low pH, which may be encountered in farms or food preparation premises, shape populations of Salmonella enterica by the selection of stress-resistant variants. METHODS AND RESULTS: Stationary-phase cultures of S. enterica serovar Enteritidis and serovar Typhimurium (one strain of each) were exposed to pH 2.5 for up to 4 h, followed by growth at pH 7 for 48 h. This process was repeated 15 times in two separate experiments, which increased the acid resistance of the three out of four populations we obtained, by three- to fourfold. Sustainable variants derived from the populations showed changes in colony morphology, expression of SEF17 fimbriae, growth, increased heat resistance and reduced virulence. CONCLUSIONS: The study demonstrates that low pH environments can select for populations of S. enterica with persistent phenotypic changes such as increased acid resistance and occasionally increased SEF17 expression and lower virulence. SIGNIFICANCE AND IMPACT OF THE STUDY: There is a common belief that increased acid resistance coincides with increased virulence. This study demonstrates for the first time that increased acid resistance often impairs virulence and affects the general phenotype of S. enterica.