Nontyphoidal Salmonella Outbreaks Associated With Chocolate Consumption: A Systematic Review.

BACKGROUND: A large, cross-border outbreak of nontyphoidal salmonellosis connected to chocolate product consumption was recently reported. This occurrence motivated us to conduct a comprehensive review of existing literature concerning outbreaks of nontyphoidal salmonellosis associated with chocolate consumption. METHODS: We performed a systematic review following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines (PROSPERO CRD42022369023) in 3 databases: U.S. National Library of Medicine, Web of Science and Excerpta Medica. Google Scholar and the bibliography of each identified report were also screened. Eligible were articles published after 1970, describing outbreaks of more than 10 patients with a nontyphoidal salmonellosis associated with chocolate consumption. RESULTS: Twenty-three articles were included, which described 12 outbreaks involving a total of 3266 patients. All outbreaks occurred in high-income countries: 1 was limited to 1 city, 6 involved 1 country and the remaining 5 involved 2 or more countries. Six outbreaks peaked in winter, 3 in autumn, 2 in spring and 1 in summer. Children were mainly affected. No predominant serotype was identified. CONCLUSIONS: Our data documents that chocolate is an optimal medium for the transmission of nontyphoidal salmonellosis. A connected worldwide reporting system including high-income, middle-income and low-income countries is crucial to detect infectious diseases outbreaks in an early phase and avoid their spread.

Recent Advances in Electrochemical Biosensors for the Detection of Salmonellosis: Current Prospective and Challenges.

Salmonellosis is a major cause of foodborne infections, caused by Salmonella, posing a major health risk. It possesses the ability to infiltrate the food supply chain at any point throughout the manufacturing, distribution, processing or quality control process. Salmonella infection has increased severely and requires effective and efficient methods for early monitoring and detection. Traditional methods, such as real-time polymerase chain reaction and culture plate, consume a lot of time and are labor-intensive. Therefore, new quick detection methods for on-field applications are urgently needed. Biosensors provide consumer-friendly approaches for quick on-field diagnoses. In the last few years, there has been a surge in research into the creation of reliable and advanced electrochemical sensors for the detection of Salmonella strains in food samples. Electrochemical sensors provide extensive accuracy and reproducible results. Herein, we present a comprehensive overview of electrochemical sensors for the detection of Salmonella by focusing on various mechanisms of electrochemical transducer. Further, we explain new-generation biosensors (microfluidics, CRISPR- and IOT-based) for point-of care applications. This review also highlights the limitations of developing biosensors in Salmonella detection and future possibilities.

Coverage of the national surveillance system for human Salmonella infections, Belgium, 2016-2020.

INTRODUCTION: The surveillance of human salmonellosis in Belgium is dependent on the referral of human Salmonella isolates to the National Reference Center (NRC). Knowledge of current diagnostic practices and the coverage of the national Salmonella surveillance system are important to correctly interpret surveillance data and trends over time, to estimate the true burden of salmonellosis in Belgium, and to evaluate the appropriateness of implementing whole-genome sequencing (WGS) at this central level. METHODS: The coverage of the NRC was defined as the proportion of all diagnosed human Salmonella cases in Belgium reported to the NRC and was assessed for 2019 via a survey among all licensed Belgian medical laboratories in 2019, and for 2016-2020 via a capture-recapture study using the Sentinel Network of Laboratories (SNL) as the external source. In addition, the survey was used to assess the impact of the implementation of culture-independent diagnostic tests (CIDTs) at the level of peripheral laboratory sites, as a potential threat to national public health surveillance programs. RESULTS: The coverage of the NRC surveillance system was estimated to be 83% and 85%, based on the results of the survey and on the two-source capture-recapture study, respectively. Further, the results of the survey indicated a limited use of CIDTs by peripheral laboratories in 2019. CONCLUSION: Given the high coverage and the limited impact of CIDTs on the referral of isolates, we may conclude that the NRC can confidently monitor the epidemiological situation and identify outbreaks throughout the country. These findings may guide the decision to implement WGS at the level of the NRC and may improve estimates of the true burden of salmonellosis in Belgium.

Exotic dried fruits caused Salmonella Agbeni outbreak with severe clinical presentation, Norway, December 2018 to March 2019.

We describe an outbreak of Salmonella Agbeni sequence type (ST)2009 infections in Norway. Between 31 December 2018 and 16 March 2019, 56 cases (33 female and 23 male; median age: 50 years, range: 2-91) were reported, of which 21 were hospitalised. Cases were defined as people living in Norway, with laboratory-confirmed infection with S. Agbeni ST2009 and cluster type (CT)2489, reported between 31 December 2018 and 30 March 2019. We conducted a case-control study, with three controls per case (matched by age, sex and municipality), using the Norwegian National Registry. Cases were more likely to have consumed a commercial mix of dried exotic fruits than controls (cases = 8, controls = 31; odds ratio: 50; 95% confidence interval: 3-2,437). The outbreak strain was confirmed by whole genome sequencing (WGS) and was isolated from the fruit mix consumed by cases, resulting in withdrawal from the market on 6 March 2019.The fruit mix consisted of fruits from different countries and continents. It was packed in Italy and distributed to several European countries, including Norway. However, no other countries reported cases. This outbreak highlights that dried fruits could represent a risk in terms of food-borne infections, which is of particular concern in ready-to-eat products.

Seasonal synchronization of foodborne outbreaks in the United States, 1996-2017.

Modern food systems represent complex dynamic networks vulnerable to foodborne infectious outbreaks difficult to track and control. Seasonal co-occurrences (alignment of seasonal peaks) and synchronization (similarity of seasonal patterns) of infections are noted, yet rarely explored due to their complexity and methodological limitations. We proposed a systematic approach to evaluate the co-occurrence of seasonal peaks using a combination of L-moments, seasonality characteristics such as the timing (phase) and intensity (amplitude) of peaks, and three metrics of serial, phase-phase, and phase-amplitude synchronization. We used public records on counts of nine foodborne infections abstracted from CDC's FoodNet Fast online platform for the US and ten representative states from 1996 to 2017 (264 months). Based on annualized and trend-adjusted Negative Binomial Harmonic Regression (NBHR) models augmented with the δ-method, we determined that seasonal peaks of Campylobacter, Salmonella, and Shiga toxin-producing Escherichia Coli (STEC) were tightly clustered in late-July at the national and state levels. Phase-phase synchronization was observed between Cryptosporidium and Shigella, Listeria, and Salmonella (ρ = 0.51, 0.51, 0.46; p < 0.04). Later peak timing of STEC was associated with greater amplitude nationally (ρ = 0.50, p = 0.02) indicating phase-amplitude synchronization. Understanding of disease seasonal synchronization is essential for developing reliable outbreak forecasts and informing stakeholders on mitigation and preventive measures.

Salmonellosis Outbreak Detected by Automated Spatiotemporal Analysis - New York City, May-June 2019.

In May 2019, the New York City Department of Health and Mental Hygiene (NYCDOHMH) detected an unusual cluster of five salmonellosis patients via automated spatiotemporal analysis of notifiable diseases using free SaTScan software (1). Within 1 day of cluster detection, graduate student interviewers determined that three of the patients had eaten prepared food from the same grocery store (establishment A) located inside the cluster area. NYCDOHMH initiated an investigation to identify additional cases, establish the cause, and provide control recommendations. Overall, 15 New York City (NYC) residents with laboratory-diagnosed salmonellosis who reported eating food from establishment A were identified. The most commonly consumed food item was chicken, reported by 10 patients. All 11 clinical isolates available were serotyped as Salmonella Blockley, sequenced, and analyzed by core genome multilocus sequence typing; isolates had a median difference of zero alleles. Environmental assessments revealed food not held at the proper temperature, food not cooled properly, and potential cross-contamination during chicken preparation. Elevated fecal coliform counts were found in two of four ready-to-eat food samples collected from establishment A, and Bacillus cereus was detected in three. The outbreak strain of Salmonella was isolated from one patient's leftover chicken. Establishing automated spatiotemporal cluster detection analyses for salmonellosis and other reportable diseases could aid in the detection of geographically focused, community-acquired outbreaks even before laboratory subtyping results become available.

An outbreak of monophasic Salmonella Typhimurium associated with raw pork sausage and other pork products, Denmark 2018-19.

In Denmark, outbreaks of salmonella with more than 20 cases have become rare. In November 2018, an outbreak of monophasic Salmonella Typhimurium was detected and an investigation initiated with the aim of identifying the source and controlling the outbreak. Outbreak cases were defined based on core genome multilocus sequence types. We conducted hypothesis-generating interviews, a matched case-control study, food sampling and trace-back investigations. We identified 49 cases distributed across Denmark. In univariable analyses a traditional form of raw Danish pork sausage (medister sausage), pork chops and ground veal/pork showed matched odds ratio of 26 (95% CI 3-207), 4 (95% CI 1-13) and 4 (95% CI 1-10), respectively. In a multivariable analysis, only medister sausage remained significant. Several patients described tasting or eating the sausage raw or undercooked. Samples of medister sausage analysed were negative for salmonella and investigations at the production site did not reveal the mechanism of contamination. In conclusion, in spite of having eliminated salmonella in the egg and broiler industry, Denmark is still at risk of major salmonella outbreaks. We identified a raw pork sausage as a particular risk product that needs to be thoroughly cooked before consumption. Tasting raw meat or eating undercooked pork should be discouraged.

Increasing incubation periods during a prolonged monophasic Salmonella Typhimurium outbreak with environmental contamination of a commercial kitchen at Oslo Airport, Norway, 2017.

In September 2017, a cluster of monophasic Salmonella Typhimurium isolates was identified at the National Reference Laboratory for Enteropathogenic Bacteria in Norway. We investigated the cluster to identify the source and implement control measures. We defined a case as a person with laboratory-confirmed salmonellosis with the outbreak strain multiple locus variable-number tandem repeat analysis type. We conducted descriptive epidemiological and environmental investigations and performed whole genome sequencing (WGS) with core and accessory genome multilocus sequence typing of all isolates from cases or the environment connected with this outbreak. We identified 21 cases, residing in 10 geographically dispersed counties, all of whom had consumed food or drinks from a café at Oslo Airport. Case distribution by date of symptom onset suggested that a point source was introduced in mid-August followed by continued environmental contamination. The incubation periods ranged 0-16 days and increased as the outbreak progressed, likely due to increasingly low-dose exposure as control measures were implemented. WGS confirmed an identical cluster type-944 in all cases and six environmental specimens from the café. Control measures, including temporary closure and kitchen refurbishment, failed to eliminate the environmental source. We recommend strengthened hygiene measures for established environmental contamination during an outbreak.

Traditional Salmonella Typhimurium typing tools (phage typing and MLVA) are sufficient to resolve well-defined outbreak events only.

Between 1991 and 2014 the per capita notification rate of salmonellosis in Australia increased from 31.9 to 69.7 cases per 100,000 people. Salmonella Typhimurium accounted for nearly half the human cases until the end of 2014. In this study, we used cluster analysis tools to compare S. Typhimurium isolates from a chicken-meat study with those reported to the National Enteric Pathogen Surveillance System (NEPSS) from the coincident human and non-human populations. There was limited phage type diversity within all populations and a lack of specificity of MLVA profiling within phage types. The chicken-meat study isolates were not significantly clustered with the human cases and at least 7 non-human sources, based on typing profiles (PT/MLVA combination), could be implicated as a source of human cases during the same period. In the absence of a strong surveillance system representative of all putative sources, MLVA and phage typing alone or in combination are insufficient to identify the source of human cases.

An international outbreak of Salmonella enterica serotype Enteritidis linked to eggs from Poland: a microbiological and epidemiological study.

BACKGROUND: Salmonella spp are a major cause of food-borne outbreaks in Europe. We investigated a large multi-country outbreak of Salmonella enterica serotype Enteritidis in the EU and European Economic Area (EEA). METHODS: A confirmed case was defined as a laboratory-confirmed infection with the outbreak strains of S Enteritidis based on whole-genome sequencing (WGS), occurring between May 1, 2015, and Oct 31, 2018. A probable case was defined as laboratory-confirmed infection with S Enteritidis with the multiple-locus variable-number tandem repeat analysis outbreak profile. Multi-country epidemiological, trace-back, trace-forward, and environmental investigations were done. We did a case-control study including confirmed and probable cases and controls randomly sampled from the population registry (frequency matched by age, sex, and postal code). Odds ratios (ORs) for exposure rates between cases and controls were calculated with unmatched univariable and multivariable logistic regression. FINDINGS: 18 EU and EEA countries reported 838 confirmed and 371 probable cases. 509 (42%) cases were reported in 2016, after which the number of cases steadily increased. The case-control study results showed that cases more often ate in food establishments than did controls (OR 3·4 [95% CI 1·6-7·3]), but no specific food item was identified. Recipe-based food trace-back investigations among cases who ate in food establishments identified eggs from Poland as the vehicle of infection in October, 2016. Phylogenetic analysis identified two strains of S Enteritidis in human cases that were subsequently identified in salmonella-positive eggs and primary production premises in Poland, confirming the source of the outbreak. After control measures were implemented, the number of cases decreased, but increased again in March, 2017, and the increase continued into 2018. INTERPRETATION: This outbreak highlights the public health value of multi-country sharing of epidemiological, trace-back, and microbiological data. The re-emergence of cases suggests that outbreak strains have continued to enter the food chain, although changes in strain population dynamics and fewer cases indicate that control measures had some effect. Routine use of WGS in salmonella surveillance and outbreak response promises to identify and stop outbreaks in the future. FUNDING: European Centre for Disease Prevention and Control; Directorate General for Health and Food Safety, European Commission; and National Public Health and Food Safety Institutes of the authors' countries (see Acknowledgments for full list).

Outbreak of Salmonella enterica serotype Poona in infants linked to persistent Salmonella contamination in an infant formula manufacturing facility, France, August 2018 to February 2019.

We describe a Salmonella Poona outbreak involving 31 infant cases in France. Following outbreak detection on 18 January 2019, consumption of rice-based infant formula manufactured at a facility in Spain was identified as the probable cause, leading to a recall on 24 January. Whole genome sequencing analysis linked present outbreak isolates to a 2010-11 S. Poona outbreak in Spain associated with formula manufactured in the same facility, indicating a persistent source of contamination.

Whole-genome sequencing of the rare Salmonella enterica serovar Anfo isolated from food handlers.

Field investigations were conducted after a small cluster of food poisoning involving six cases was reported. While no stool samples were available from the cases for microbiological testing, Salmonella species was found to be present in the stools of food handlers with gastroenteritis symptoms. Four Salmonella isolates recovered from the food handlers were retrospectively investigated at the genome level using whole-genome sequencing (WGS). WGS showed that S. Anfo (antigenic formulae 39:y:1,2), a rarely isolated serovar, caused infections in the food handlers. S. Anfo analysed in this study contained virulence factors required for causing disease. They did not contain any antibiotic resistance genes or plasmid. The epidemiologically related isolates differed to each other by a maximum of one single nucleotide polymorphism. WGS was useful in identifying rare Salmonella serovars and it is potentially more cost-effective than traditional serotyping methods. It can also confidently group epidemiologically related isolates belonging to S. Anfo.

Salmonellosis in Poland in 2017

AIM: The aim of the study is to assess the epidemiological situation of salmonellosis in 2017 in Poland compared to the previous years. MATERIAL AND METHODS: The assessment of the epidemiological situation was performed on the basis of data submitted to the Department of Epidemiology of Infectious Diseases and Surveillance NIPH-NIH by sanitaryepidemiological stations by means of the Epidemiological Case Reporting System (SRWE) and the Registry of Epidemic Outbreaks (ROE), along with data from the annual bulletin "Infectious diseases and poisonings in Poland in 2017" (NIPH-NIH, CSI, Warsaw 2018), and information from the laboratories of Sanitary epidemiological Stations as well as data from the Department of Demographic Studies of the Central Statistical Office. RESULTS: In 2017, 10 000 cases of Salmonella infection were registered in Poland, 9 710 were cases of intestinal salmonellosis, 290 were forms of extraintestinal salmonellosis. The incidence rate per 100 thousand population was 26.0. The hospitalization rate for all salmonellosis cases was 63.3%. Confirmed cases accounted for 92,1% of all cases, the remaining 7.9% were probable cases. Peak incidence occurred, as in previous years, in the summer months. The voivodships with the largest number of cases caused by Salmonella were the Mazowieckie and Malopolskie voivodeships, the least was recorded in the Lubuskie voivodeship. The age group in which the highest percentage of food poisonings was recorded were children aged 0-4, while extraintestinal forms most often concerned people aged over 60. 278 food poisoning outbreaks were recorded, in which Salmonella Enteritidis was the most frequently isolated serotype. This serotype was most often isolated in foodborne outbreaks and in sporadic cases, it is responsible for 92% of all salmonellosis. Salmonella infection was found in 0.2% of people working with food and in 8.1% of contacts of the cases. According to data from the Central Statistical Office, 10 people died of salmonellosis in 2017. CONCLUSIONS: In 2017, the number of people infected with Salmonella was as high as in 2016, the reason for the persistence of such a high number of cases may be the association of some infections with an international salmonellosis outbreak, the peak of which was in 2016. The high incidence of salmonellosis may also be a consequence of legal changes introduced in 2014 regarding the reporting of positive test results by laboratories for epidemiological surveillance.

A film-based integrated chip for gene amplification and electrochemical detection of pathogens causing foodborne illnesses.

Given the increased interest in public hygiene due to outbreaks of food poisoning, increased emphasis has been placed on developing novel monitoring systems for point-of-care testing (POCT) to evaluate pathogens causing foodborne illnesses. Here, we demonstrate a pathogen evaluation system utilizing simple film-based microfluidics, featuring simultaneous gene amplification, solution mixing, and electrochemical detection. To minimize and integrate the various functionalities into a single chip, patterned polyimide and polyester films were mainly used on a polycarbonate housing chip, allowing simple fabrication and alignment, in contrast to conventional polymerase chain reaction, which requires a complex biosensing system at a bench-top scale. The individual integrated sensing chip could be manually fabricated in 10 min. Using the developed film-based integrated biosensing chip, the genes from the pathogens causing foodborne illnesses were simultaneously amplified based on multiple designed microfluidic chambers and Hoechst 33258, which intercalates into double-stranded DNA, to generate the electrochemical signal. The target pathogen gene was accurately analyzed by square wave voltammetry (SWV) within the 25 s, while the gel electrophoresis required about 30 min. Based on the developed integrated biosensing chip, the 1.0 × 101 and 1.0 × 102 colony-forming unit (CFU) of Staphylococcus aureus and Escherichia coli were sensitively detected with high reproducibility in the 25 s. On the basis of the significant features of the film-based molecular analysis platform, we expect that the developed sensor could be applied to the screening of various pathogens as a POCT device.

Highly suspected cases of salmonellosis in two cats fed with a commercial raw meat-based diet: health risks to animals and zoonotic implications.

BACKGROUND: Feeding raw meat-based diets (RMBD) to companion animals raises public health concerns for both animals and humans. While considerable attention has been paid to bacterial contamination of commercial pet food, few literature studies have investigated foodborne disease in companion animals. Salmonellosis is reported to be infrequent in cats but no known data or studies estimating feline salmonellosis are available or large-scale epidemiological studies assessing Salmonella risk factors. CASE PRESENTATION: Two highly suspected cases of salmonellosis in two cats fed with a commercial frozen poultry RMBD are presented, for the first time from the same household. The clinical presentation, diagnostics, treatment and follow-up are reported and the zoonotic implications are discussed. CONCLUSIONS: This case highlights the health risks posed to both animals and owners by feeding RMBD to pets, and suggests that these risks should be considered by veterinary practitioners.

An outbreak of Salmonella Saintpaul gastroenteritis after attending a school camp in the Northern Territory, Australia.

An outbreak of salmonellosis occurred following attendance at a school camp between 5 and 8 August 2014 in a remote area of the Northern Territory, Australia. We conducted a retrospective cohort study via telephone interviews, using a structured questionnaire that recorded symptoms and exposures to foods and activities during the camp. A case was anyone with laboratory confirmed Salmonella Saintpaul infection or a clinically compatible illness after attending the camp. Environmental health officers from the Environmental Health Branch undertook an investigation and collected water and environmental samples. We interviewed 65 (97%) of the 67 people who attended the camp. There were 60 students and 7 adults. Of the 65 people interviewed, 30 became ill (attack rate 46%); all were students; and 4 had laboratory confirmed S. Saintpaul infection. The most commonly reported symptoms were diarrhoea (100% 30/30), abdominal pain (93% 28/30), nausea (93% 28/30) and fever (70% 21/30). Thirteen people sought medical attention but none required hospitalisation. Illness was significantly associated with drinking cordial at lunch on 7 August (RR 3.8, 95% CI 1.3-11, P < 0.01), as well as drinking cordial at lunch on 8 August (RR 2.1, 95% CI 1.1-4.2, P=0.01). Salmonella spp. was not detected in water samples or wallaby faeces collected from the camp ground. The epidemiological investigation suggests the outbreak was caused by environmental contamination of food or drink and could have occurred during ice preparation or storage, preparation of the cordial or from inadequate sanitising of the cooler from which the cordial was served. This outbreak highlights the risks of food or drink contamination with environmental Salmonella. Those preparing food and drink in campground settings should be vigilant with cleaning, handwashing and disinfection to prevent outbreaks of foodborne disease.

An outbreak of salmonellosis associated with duck prosciutto at a Northern Territory restaurant.

In June 2015, an outbreak of salmonellosis occurred among people who had eaten at a restaurant in Darwin, Northern Territory over 2 consecutive nights. We conducted a retrospective cohort study of diners who ate at the restaurant on 19 and 20 June 2015. Diners were telephoned and a questionnaire recorded symptoms and menu items consumed. An outbreak case was defined as anyone with laboratory confirmed Salmonella Typhimurium PT9 (STm9) or a clinically compatible illness after eating at the restaurant. Environmental health officers inspected the premises and collected food samples. We contacted 79/83 of the cohort (response rate 95%); 21 were cases (attack rate 27%), and 9 had laboratory confirmed STm9 infection. The most commonly reported symptoms were diarrhoea (100%), abdominal pain (95%), fever (95%) and nausea (95%). Fifteen people sought medical attention and 7 presented to hospital. The outbreak was most likely caused by consumption of duck prosciutto, which was consumed by all cases (OR 18.6, CI 3.0-∞, P < 0.01) and was prepared on site. Salmonella was not detected in any food samples but a standard plate count of 2 x 107 colony forming units per gram on samples of duck prosciutto demonstrated bacterial contamination. The restaurant used inappropriate methodology for curing the duck prosciutto. Restaurants should consider purchasing pre-made cured meats, or if preparing them on site, ensure that they adhere to safe methods of production.

The Epidemiology of Childhood Salmonella Infections in Alberta, Canada.

OBJECTIVES: The objectives were to describe the incidence, demographics, laboratory findings, and suspected sources of childhood Salmonella infections in Alberta, Canada, with a focus on preventable cases. METHODS: Data from Notifiable Disease Reports for children with nontyphoidal salmonellosis (NTS) or typhoid/paratyphoid fever from 2007 through 2015 were analyzed. RESULTS: NTS was detected from 2285 children. Bacteremia was documented in 55 cases (2.4%), whereas a single infant had NTS meningitis. The suspected source was food (N = 577; 25.3%) followed by animal or animal manure contact (N = 426; 18.6%), of which a reptile was the suspected source in 264 cases (11.5%). There were 44 outbreaks with none sharing the same food source. Ninety-five children were diagnosed with typhoid/paratyphoid fever, of which 48 cases (51%) were typhoid cases in unimmunized children 2 years or older. CONCLUSIONS: There are still ∼275 pediatric cases of Salmonella infection in Alberta annually, the bulk of which are preventable. APPLICATION: Public education about reptile exposure, food safety, and pretravel immunizations could potentially prevent many cases of Salmonella infection.

Typhoid Fever on the Half Shell.

Protecting the public from communicable infectious disease outbreaks is one of the most important, and most challenging, functions of public health. Foodborne outbreaks are not uncommon, and they can be especially difficult. This true story of the epidemiologic investigation into a typhoid fever outbreak illustrates the critical importance of timely reporting by front-line clinicians, extensive interprofessional teamwork, and statewide coordination.