Category Archives: WGS

Research – Estimating Underdetection of Foodborne Disease Outbreaks

CDC

To the Editor: In the February issue, Ford et al. used the power law to estimate underdetection of foodborne disease outbreaks in the United States (1). Two of their main conclusions are entirely reasonable: small outbreaks are more likely to go undetected than large outbreaks, and the use of whole-genome sequencing (WGS) has improved the detection of small outbreaks caused by pathogens for which WGS is used. However, their conclusion on the usefulness of the power law itself needs further consideration.

Ford et al. analyzed the size of all foodborne outbreaks reported to the national Foodborne Disease Outbreak Surveillance System during 1998–2019. They defined outbreak size as the number of laboratory-confirmed cases. However, laboratory-confirmed cases are only good estimators for the size of outbreaks detected through pathogen-specific surveillance, such as for Salmonella, where outbreak detection follows the accumulation of confirmed cases. For outbreaks associated with events or establishments, identification might rely on reports from consumers, many of whom do not seek healthcare; thus, stool specimens might only be collected from a few cases to confirm the etiology. Consumer complaints are the primary means for identifying foodborne outbreaks caused by norovirus. The Council to Improve Foodborne Outbreak Response recommends collecting clinical specimens from >5 members from the ill group in such settings (2). Thus, the number of confirmed cases in an outbreak is dependent on how the outbreak is detected. Outbreaks detected by complaint generally have few confirmed cases, even though they can involve large numbers of illnesses.

To provide a fair evaluation for the usefulness of the power law, it may be better to restrict analyses to outbreaks with common detection pathways. For outbreaks detected by pathogen-specific surveillance, counting confirmed cases seems appropriate. For outbreaks detected through consumer complaints, analyses should include all outbreak-associated illnesses.

Research – Shiga Toxin-Producing Escherichia coli Strains from Romania: A Whole Genome-Based Description

MDPI

Abstract

The zoonotic Shiga toxin-producing Escherichia coli (STEC) group is unanimously regarded as exceptionally hazardous for humans. This study aimed to provide a genomic perspective on the STEC recovered sporadically from humans and have a foundation of internationally comparable data. Fifty clinical STEC isolates, representing the culture-confirmed infections reported by the STEC Reference Laboratory between 2016 and 2023, were subjected to whole-genome sequencing (WGS) analysis and sequences were interpreted using both commercial and public free bioinformatics tools. The WGS analysis revealed a genetically diverse population of STEC dominated by non-O157 serogroups commonly reported in human STEC infections in the European Union. The O26:H11 strains of ST21 lineage played a major role in the clinical disease resulting in hospitalisation and cases of paediatric HUS in Romania surpassing the O157:H7 strains. The latter were all clade 7 and mostly ST1804. Notably, among the Romanian isolates was a stx2a-harbouring cryptic clade I strain associated with a HUS case, stx2f– and stx2e-positive strains, and hybrid strains displaying a mixture of intestinal and extra intestinal virulence genes were found. As a clearer picture emerges of the STEC strains responsible for infections in Romania, further surveillance efforts are needed to uncover their prevalence, sources, and reservoirs.

UK – Investigation into an outbreak of Shiga toxin-producing E. coli (STEC) O145 in Great Britain, May to June 2024

Gov UK

The UK Health Security Agency (UKHSA), Public Health Scotland, Public Health Wales and Public Health Agency Northern Ireland (PHA), in collaboration with the Food Standards Agency (FSA) and Food Standards Scotland (FSS) have been working together with local authorities to investigate an outbreak of Shiga toxin-producing Escherichia coli (STEC) O145 identified through the analysis of whole genome sequencing (WGS) data in May 2024.

A potential outbreak was first identified in England on 22 May 2024 through UKHSA’s routine surveillance, with a rapid ten-fold increase in the number of faecal samples from patients testing positive for non-O157 STEC toxin genes referred from the NHS to the national reference laboratory.

On 24 May, reference laboratory polymerase chain reaction (PCR) test results indicated the increase was likely driven by a strain of STEC which possessed the stx2a, eae+ virulence gene profile but was neither serotype O157 or O26 (defined as ‘probable cases’) and increased hospital emergency department attendances for gastrointestinal illness (1). Concurrent increases for similar surveillance indicators were reported in Wales and Scotland. A national incident was declared on 24 May.

Subsequent WGS analysis available on 28 May showed that the majority of these probable cases had illness caused by STEC serotype O145 with a specific genetic profile (‘genetic fingerprint’). These confirmed outbreak cases with a 5-single nucleotide polymorphism (SNP) termed ‘t5.206’, were distinct from other STEC O145 outbreaks recently investigated and from sporadic cases. This was a re-emergence of a STEC cluster investigated in 2023, where no source could be conclusively confirmed. No international cases of a similar genotype were reported on global databases and information was shared through established international communication platforms with other countries.

Between 25 May and 24 June there were 275 confirmed cases reported (273 primary cases and 2 secondary cases (Figure 1)). Cases were geographically dispersed in all regions of England (182), in Scotland (58), Wales (31) and Northern Ireland (4), with no notable geographic clustering. However, evidence suggests that cases resident in Northern Ireland likely acquired their infection in England. Primary cases were predominantly female (57%) and had a median age of 30 years (range: 1 to 89 years). The most affected age groups were 20 to 29 and 30 to 39 with 30% and 23% of cases respectively.

The STEC O145 t5:206 outbreak strain possesses a virulence gene profile (stx2a, eae+) which is associated with more severe disease and an increased likelihood of Haemolytic Uraemic Syndrome (HUS), a clinical syndrome associated with STEC which can lead to kidney failure and death.

Among confirmed t5:206 outbreak cases (Table 1):

  • 81% have reported bloody diarrhoea
  • 49% of cases have been admitted to hospital
  • 10% attended A&E for their symptoms

Of the 122 hospitalised cases, 57% were female and had a median age of 35 (with a range of 6 to 85 years).

To date 7 cases of HUS have been confirmed to be associated with this outbreak. Of the 7 HUS reported cases:

  • 5 were resident in England
  • 2 were resident in Scotland
  • their ages ranged from 12 to 59
  • the majority were female (n=6)

These HUS cases all reported onset of illness in mid to late May. There have been 2 reported deaths in patients within 28 days of confirmation of infection with the STEC outbreak strain t5:206. Neither of these patients were diagnosed with HUS, both were adults and had underlying medical conditions. Based on the information available, one of these deaths is likely linked to STEC infection.

Early epidemiological analyses indicated that the cases were widely geographically dispersed across the UK with a rapid increase in confirmed cases consistent with a nationally distributed food product as the cause of the outbreak. The FSA and FSS therefore were involved from the start of the investigation. In the initial phase of the investigation, analysis of available information on reported cases, including routinely collected case questionnaire data (which captures clinical symptoms and environmental plus a wide range of food exposures (2), indicated a higher than expected proportion of cases reported consuming pre-packaged sandwich products during the 7 days prior to becoming unwell.

The hypothesis that sandwiches containing lettuce were the likely main cause of the outbreak was confirmed through multiple different epidemiological studies which showed a statistically significant association between illness and consumption of these products.

Food chain investigations were carried out by FSA and FSS, informed by the food histories of cases and alongside the epidemiological investigations carried out by the UK public health agencies. Lettuce was the main focus as the likely contaminated sandwich ingredient. The FSA and FSS initially identified one supplier of the potentially contaminated lettuce, investigations are still ongoing at 2 other lettuce suppliers at the grower stage of the supply chain.

Further investigations carried out by FSA and FSS with the identified sandwich product producers and salad growers indicated that these food business operators (FBOs) had detailed and robust hazard analysis and critical control points (HACCP) plans and biosecurity in place. Enhanced testing at these FBOs was carried out in response to the investigation. While all test results were negative for STEC, general or ‘indicator’ E. coli was identified in sandwich and lettuce products, indicating a possible contamination event had occurred. As a result of the epidemiological and food chain investigation outcomes, the FBOs elected to undertake a voluntary withdrawal and recall of all potentially associated ready to eat products (sandwiches, wraps and salads) – first on 14 June and then, following further findings, on the 15 and 16 June (345) . Investigations into the root cause of the outbreak are still ongoing.

There are challenges in the investigation of any foodborne disease outbreak of STEC as previously reported (6). In the case of short shelf life and highly perishable products such as salad products, microbiological confirmation of the outbreak strain (the same WGS profile) in the foods implicated in the outbreak is very difficult as the foods have usually already been consumed and there is no residual product to test. As performed in this investigation, environmental and food testing may be useful for the detection of E. coli as an indicator of a potential contamination event, as well as detection of specific toxin genes present in STEC which causes food poisoning.

Rapid response based on early surveillance indicators prior to an outbreak being confirmed, and subsequent rapid confirmation of the outbreak (via routine implementation of WGS for characterisation of STEC and other major bacterial pathogen causes of foodborne disease) is crucial for the protection of public health. Early collaboration and communication with UK government agencies and international counterparts identified that the outbreak was limited to the UK, supporting the epidemiological and food chain investigations findings that a UK produced salad product was the cause of the outbreak.

STEC is a zoonotic pathogen (meaning a bacteria transmissible between humans and animals) and is naturally present in the intestines of animals, especially ruminant animals such as cattle and sheep. It is not always possible to determine the reasons for or causes of contamination of food products with zoonotic pathogens that can cause food poisoning outbreaks. Ready-to-eat salad vegetables can be contaminated with pathogens at the pre-harvest level (via flooding, rainwater run-off or irrigation water containing animal faeces) or post-harvest during washing and packaging.

As of 24 June, newly reported confirmed cases have now markedly declined (Figure 1) and the surveillance indicators for probable cases have now reduced back down to expected levels for this time of year. The majority of the recently reported probable cases are not part of the O145 t5:206 outbreak, but rather are small numbers of sporadic cases. A small number of cases may be reported in the short term due to the time lag between:

  • when people become ill
  • when they visit their GP or hospital
  • when a sample is taken for testing
  • when this sample is referred to the national reference laboratories for further typing
  • the availability of WGS results

Based on available data, the ongoing public health risk is minimal.

Public health agencies are continuing to monitor surveillance and WGS data for confirmed cases and follow up cases to identify any common links. The FSA and FSS continue to work with the relevant local authorities, salad growers, sandwich suppliers and manufacturers to identify the root cause of the outbreak so that actions can be taken to prevent a re-occurrence.

Research – Prolonged multi-country outbreak of Listeria monocytogenes ST173 linked to consumption of fish products 2012-2024

EFSA

Abstract

Between 2012 and 2024, a prolonged outbreak of 73 cases of L. monocytogenes ST173 infections (‘My2’ -cluster) has been ongoing in Belgium (5), Czechia (1), Germany (39), Finland (2), Italy (1), the Netherlands (20), and the United Kingdom (UK) (5). Fourteen deaths have been recorded as associated with this outbreak. Males aged over 60 years represent the most affected population group. Most patients with available information from case interviews report consuming various fish products before illness. Among 73 cases, an ongoing ‘My2’ sub-cluster 1 was identified with 38 cases reported in six EU countries and the UK.

The ‘My2’ sub-cluster 1 includes 83 non-human isolates, of which 48 are food isolates and six are environmental isolates with traceability data corresponding to 37 fish products (different typologies) and 12 fish manufacturers (located in six EU countries and one third country). The epidemiological and traceability information did not allow identification of a single common point or food business operator in the traceability of all products. Five countries reported the implementation of control measures.

The whole genome analysis (WGS) analysis, the tracing evidence, and the detection in fish products between 2017 and 2024 suggest that the strain has been geographically spread in Europe for several years, most likely originated from a past single source high up in the production chain, and has been established in different fish processing plants. Overall, the evidence supports the hypothesis of fish products as the vehicle of infections.

Given the detection of the outbreak strain in multiple types of fish products and its wide circulation in the EU/EEA fish production chain, new cases are likely to be reported. Corrective measures should be implemented in the plants where the contamination is detected, and further investigations should be carried out to identify the point of entry and prevent possible re-contamination.

Research -Investigation of outbreak of multidrug-resistant Salmonella shows link to pet treats

Food Safety News

A new study investigating a multistate outbreak of multidrug-resistant Salmonella infections linked to pig ear pet treats highlights the interconnectedness of human health and pet ownership, emphasizing the need for stringent surveillance of pet food products.

The study was funded by the Food and Drug Administration’s Animal Feed Regulatory Program Standards and the FDA’s Veterinary Laboratory Investigation and Response Network (Vet-LIRN).

From June 2015 to Sept. 2019, 154 human cases of Salmonella infection were reported across 34 states. The investigation identified seven Salmonella serotypes genetically related to samples from pig ear pet treats. Whole genome sequencing (WGS) of isolates was used to predict antimicrobial resistance. Notably, 107 of 122 patients interviewed reported dog contact, and 65 of 97 reported contact with pig ear pet treats. Salmonella was isolated from 137 pig ear treats, including imports from Argentina, Brazil, and Colombia, and from four dogs. WGS predicted that 77 percent of human isolates and 43 percent of pig ear treat isolates were resistant to three or more antimicrobial classes.

Research – Virulence Potential and Antimicrobial Resistance of Listeria monocytogenes Isolates Obtained from Beef and Beef-Based Products Deciphered Using Whole-Genome Sequencing

MDPI

Abstract

Listeria monocytogenes is a ubiquitous bacterial pathogen that threatens the food chain and human health. In this study, whole-genome sequencing (WGS) was used for the genomic characterization of L. monocytogenes (n = 24) from beef and beef-based products. Multilocus Sequence Type (MLST) analysis revealed that ST204 of CC204 was the most common sequence type (ST). Other sequence types detected included ST1 and ST876 of CC1, ST5 of CC5, ST9 of CC9, ST88 of CC88, ST2 and ST1430 of CC2, and ST321 of CC321. Genes encoding for virulence factors included complete LIPI-1 (pfrAhlyplcAplcBmplactA) from 54% (13/24) of the isolates of ST204, ST321, ST1430, and ST9 and internalin genes inlABC that were present in all the STs. All the L. monocytogenes STs carried four intrinsic/natural resistance genes, fosXlinnorB, and mprF, conferring resistance to fosfomycin, lincosamide, quinolones, and cationic peptides, respectively. Plasmids pLGUG1 and J1776 were the most detected (54% each), followed by pLI100 (13%) and pLM5578 (7%). The prophage profile, vB_LmoS_188, was overrepresented amongst the isolates, followed by LP_101, LmoS_293_028989, LP_030_2_021539, A006, and LP_HM00113468. Listeria genomic island 2 (LGI-2) was found to be present in all the isolates, while Listeria genomic island 3 (LGI-3) was present in a subset of isolates (25%). The type VII secretion system was found in 42% of the isolates, and sortase A was present in all L. monocytogenes genomes. Mobile genetic elements and genomic islands did not harbor any virulence, resistance, or environmental adaptation genes that may benefit L. monocytogenes. All the STs did not carry genes that confer resistance to first-line antibiotics used for the treatment of listeriosis. The characterization of L. monocytogenes in our study highlighted the environmental resistance and virulence potential of L. monocytogenes and the risk posed to the public, as this bacterium is frequently found in food and food processing environments.

Research- WGS Responsible for Quick Detection of Salmonella Outbreak

IDSE

The use of whole genome sequencing (WGS)-based surveillance and rapid epidemiological intervention allowed public health experts to quickly link a Salmonella enterica serovar Typhimurium outbreak to cantaloupes grown in the United States.

Research – Uncommon Salmonella Infantis Variants with Incomplete Antigenic Formula in the Poultry Food Chain, Italy

CDC

Uncommon Salmonella Infantis variants displaying only flagellar antigens phenotypically showed identical incomplete antigenic formula but differed by molecular serotyping. Although most formed rough colonies, all shared antimicrobial resistances and the presence of usg gene with wild-type Salmonella Infantis. Moreover, they were undistinguishable wild-type Salmonella Infantis by whole-genome sequencing.

USA – E. coli Outbreak linked to PCC Guacamole

Marler Blog

Public Health is investigating an outbreak of Shiga toxin-producing E. coli O157:H7 (also known as STEC). Three of the five people who got sick had testing that matched by genetic fingerprinting. This means they most likely got sick from the same source. Based on information collected, we found one common source for all sick people which was a store-made guacamole purchased at PCC Community Markets – West Seattle Co-op on February 10, 2024. Even though we think this is the most likely source of illness, we do not know for certain. We did not find out how the guacamole might have been contaminated with STEC and did not have any left to test. This outbreak appears to be over.

Research- Notes from the Field: Rapidly Linking an Outbreak of Salmonella Typhimurium Infections to Domestically Grown Cantaloupes Through Early Collaboration — United States, 2022

CDC

Summary

What is already known about this topic?

A 2020 outbreak of Salmonella infections was found to be associated with melons after conclusion of harvesting, when melons were no longer likely to be on the market.

What is added by this report?

In 2022, whole genome sequencing (WGS)–based Salmonella surveillance, historical melon farm environmental sampling results, and patient interviews were used to rapidly link a Salmonella Typhimurium outbreak to contaminated cantaloupes.

What are the implications for public health practice?

WGS-based surveillance, combined with rapid collection of epidemiologic data by state and local agencies, can be used to reduce the time to outbreak detection and response.