Outbreak of Salmonella Typhimurium linked to Swedish pre-washed rocket salad, Sweden, September to November 2022.

In September 2022, the Public Health Agency of Sweden observed an increase in domestic Salmonella Typhimurium cases through the Swedish electronic notification system, and an outbreak strain was identified with whole genome sequencing. Overall, 109 cases with symptom onset between 17 September and 24 November 2022 were reported from 20 of 21 Swedish regions. The median age of cases was 52 years (range 4-87 years) and 62% were female. A case-control study found cases to be associated with consumption of rocket salad (adjusted odds ratio (aOR) = 4.9; 95% confidence interval (CI): 2.4-10, p value < 0.001) and bagged mixed salad (aOR = 4.0; 95% CI: 1.9-8.1, p value < 0.001). Trace-back, supported by Finnish authorities who identified the Swedish outbreak strain in a Finnish cluster during the same time period, identified rocket salad, cultivated, pre-washed and pre-packed in Sweden as the likely source of the outbreak. No microbiological analyses of rocket salad were performed. Our investigation indicates that bagged leafy greens such as rocket salad, regardless of pre-washing procedures in the production chain, may contain Salmonella and cause outbreaks, posing a health risk to consumers. We emphasise the need for primary producers of leafy greens to identify possible contamination points to prevent outbreaks.

Occurrence and Characteristics of Toxigenic Staphylococcus aureus in Retail Foods in Iran.

Staphylococcus aureus causes various toxigenic and invasive diseases in humans worldwide. This study examined the prevalence, virulence genes, and antibiotic resistance of S. aureus isolates collected from 894 retail food samples in Ardabil, Iran. Staphylococcal cassette chromosome mec (SCCmec), spa, and multilocus sequence typing methods were employed to further investigate the molecular characteristics of methicillin-resistant S. aureus (MRSA) isolates. The results revealed that 11.18% (n = 100) of food samples exhibited contamination with S. aureus (10.50% methicillin-sensitive S. aureus [MSSA] and 0.67% MRSA). Notably, raw minced meat (29.41%), Faloodeh (25%), and Olivier salad (21.42%) emerged as the most frequently contaminated food items. Among the 100 isolates of S. aureus, 94% were characterized as MSSA, with the remaining 6% identified as MRSA. The highest resistance was observed for penicillin (12%). MRSA isolates exhibited significantly higher resistance rates. Seventy-nine percent of the isolates were positive for sea, 14% for seb, 8% for a sec, and 0% for sed enterotoxin-encoding genes. Sixteen percent of isolates harbored two or more staphylococcal enterotoxin genes, simultaneously. Moreover, 97%, 94%, 24%, and 22% of isolates were positive for hla, hld, tst, and pvl virulence-encoding genes, respectively. No isolate was positive for the exfoliative toxins encoding eta and etb genes. MRSA isolates belonged to CC8 (n = 4) and CC22 (n = 2). Isolates in CC8 belonged to lineage ST239-MRSA-III and spa type t030; the isolates in CC22 belonged to ST22-MRSA-IV and spa types t310 and t223. In conclusion, a relatively high proportion of our retail food samples were contaminated with S. aureus. The high incidence of isolates with toxigenic genes raises serious health concerns. Furthermore, the presence of MRSA lineages linked to humans suggests that retail foods may be contaminated with human origin.

Prevalence, antimicrobial susceptibility test and associated factors of Salmonella and Shigella in ready-to-eat fruit juices and salads in Mekelle, northern Ethiopia.

BACKGROUND: Food borne diseases is a challenging problem nowadays. Salmonella and Shigella species are great concern of food-born outbreaks. Thus, this study was aimed to assess the prevalence, antimicrobial susceptibility test and associated factors of Salmonella and Shigella species in fruit juices and salads. METHODS: A community based cross sectional study design was carried out on 50 juice houses from December to March 2020 in Mekelle. One hundred fifty samples were collected aseptically from the juice houses for laboratory analysis. Information related to risk factors was obtained using a structured questionnaire. In the laboratory, samples were homogenized using peptone water and incubated overnight for enrichment. Then, Salmonella and Shigella species were isolated on Salmonella-Shigella agar and Xylose Lysine Deoxycholate agar. Disc diffusion method was used to perform antimicrobial susceptibility test. Using SPSS (version 22) package, descriptive statistics and Chi square test (χ2) were used to analyze the data, and p < 0.05 was considered as statistically significant. RESULT: The overall prevalence of Salmonella and Shigella species was 41/150 (27.33%; 95% CI: 20.20 - 34.46) with 33 (22%) Salmonella spp. and 8(5.33%) Shigella spp. Antimicrobial susceptibility tests of both Salmonella and Shigella spp.showed high resistance against ampicillin (100%), tetracycline (63.6 and 62.5%, respectively) and amoxicillin-clavulanic acid (100%). Accessibility of fruits to flies and dust had statistical association (p = 0.021) with occurrence of Salmonell a and/or Shigella spp. CONCLUSION: The overall prevalence of Salmonella and Shigella spp. was found to be significant. The resistant rate of isolates against ampicillin, tetracycline and amoxicillin-clavulanic acid was high. Storage sites for fruits should be inaccessible to flies and dust. Therefore, routine monitoring of juice houses should be promoted and regular evaluation of bacterial resistance pattern should be done for selective antimicrobial therapy. Furthermore, consistent training of juice makers on food safety and hygiene should be implemented by the concerned body.

School Nutrition Personnel Perceptions of School Salad Bars before and after COVID-19.

Many schools have salad bars as a means to increase students' fruit and vegetable intake. School nutrition programs experienced drastic changes to the school food environment due to COVID-19. The aim of the current study was to understand cafeteria personnel's experiences related to salad bar implementation before the COVID-19 pandemic and in the current school environment to inform efforts to enhance salad bar sustainability. Seven elementary schools (N = 30 personnel) installed salad bars prior to COVID-19; three of these schools (n = 13 personnel) re-opened salad bars after COVID-19. Cafeteria personnel completed surveys assessing their experiences with salad bars at both time points. Satisfaction with salad bar implementation and training was high pre- and post-COVID-19. Most agreed that salad bars increased students' fruit and vegetable intake, yet had concerns about cleanliness and waste. Perceived job difficulty increased post-COVID-19 (p = 0.01), and satisfaction with student salad bar training decreased (p = 0.001). Additional staff support and greater student training were needed post-COVID-19. Overall, salad bars were viewed favorably; however, more challenges and lower satisfaction were reported following COVID-19. Increasing support for cafeteria personnel is needed for salad bar sustainability and improving the school food environment.

Effect of Gamma Irradiation on Pathogenic Staphylococcus aureus in Packaged Ready-to-Eat Salads Treated with Biological Extracts.

Providing pathogen-free ready-to-eat (RTE) salads is critical for all consumers, especially individuals with weakened immunity. In this study, the efficacy of γ-irradiation on Staphylococcus aureus (S. aureus) in freshly packaged salads (4.24 log CFU/g) treated with essential oil (EO) and myrtle juice during 10 days of storage and their impact on organoleptic properties were investigated. EO was extracted by hydrodistillation and the chemical composition was analyzed by gas chromatography with Flame Ionization Detector (GC/FID) and gas chromatography/mass spectrometry (GC/MS). Myrtle juice was prepared from fresh fruits. The cytotoxic effects of Thymus capitatus (T. capitatus) EO against a normal human umbilical vein endothelial cell (HUVEC) were assessed. GC/FID and GC-MS analysis of the thyme EO revealed the presence of 13 compounds, including carvacrol (79.55%) and p-cymene (7.93%) as major components. The EO was found to be noncytotoxic, with concentrations lower than 0.16 µL/mL. A reduction of more than 3 log CFU/g and a total inactivation of S. aureus were achieved with the combination of gamma irradiation at 0.5 kGy with myrtle juice at 6 µL/mL and EO at 0.08 µL/mL, respectively. The treatment of fresh RTE salads with thyme and myrtle juice was evaluated as acceptable by the sensory panel. The combined effect showed a synergistic potential on the inactivation of S. aureus.

The use of active compounds to shape the quality of active double-layer films based on furcellaran intended for packaging salad-dressing - Assessment of utilitarian and storage properties.

In order to obtain innovative, high-quality biodegradable packaging, double-layer films based on furcellaran and gelatin, enriched with plant extracts were created. The films were assessed considering their potential utility, applications and environmental impact. The mechanical properties over a period of nine months were studied and it was noted that the passing of time had a beneficial effect on these parameters. The antioxidant properties was also examined, with the highest results obtained using the DPPH and metal chelating activity methods for GE (76.64 % and 9.85 % respectively), while this film showed the lowest FRAP value (5.99 %) compared to the highest obtained for DTE (52.62 %). For the first time, the possibility of using the double-layer active FUR/GEL film as packaging for salad-dressing was evaluated, but no improvement in parameters was observed regardless of the extract used. The environmental impact analysis showed the ability to completely decomposed in vermicompost within several days.

Application of High-Resolution Melting and DNA Barcoding for Discrimination and Taxonomy Definition of Rocket Salad (Diplotaxis spp.) Species.

Nuclear and cytoplasmic DNA barcoding regions are useful for plant identification, breeding, and phylogenesis. In this study, the genetic diversity of 17 Diplotaxis species, was investigated with 5 barcode markers. The allelic variation was based on the sequences of chloroplast DNA markers including the spacer between trnL and trnF and tRNA-Phe gene (trnL-F), the rubisco (rbcl), the maturase K (matk), as well as the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA. A highly polymorphic marker (HRM500) derived from a comparison of cytoplasmic genome sequences in Brassicaceae, was also included. Subsequently, a real-time PCR method coupled with HRM analysis was implemented to better resolve taxonomic relationships and identify assays suitable for species identification. Integration of the five barcode regions revealed a grouping of the species according to the common chromosomal set number. Clusters including species with n = 11 (D. duveryrieriana or cretacea, D. tenuifolia, D. simplex and D. acris), n = 8 (D. ibicensis, D. brevisiliqua and D. ilorcitana), and n = 9 (D. brachycarpa, D. virgata, D. assurgens, and D. berthautii) chromosomes were identified. Both phylogenetic analysis and the genetic structure of the collection identified D. siifolia as the most distant species. Previous studies emphasized this species' extremely high glucosinolate content, particularly for glucobrassicin. High-resolution melting analysis showed specific curve patterns useful for the discrimination of the species, thus determining ITS1 as the best barcode for fingerprinting. Findings demonstrate that the approach used in this study is effective for taxa investigations and genetic diversity studies.

Performance of a manually operated salad spinner centrifuge for serum separation in the healthy domestic horse (Equus caballus) and southern white rhinoceros (Ceratotherium simum).

BACKGROUND: Field veterinarians and researchers studying wild species, such as the southern white rhinoceros, often work in remote areas with limited access to standard laboratory equipment, hindering the ability to measure serum analytes. OBJECTIVES: The first objective was to produce an inexpensive, manually operated centrifuge that could accept standard laboratory tubes by modifying a consumer-grade salad spinner with low-cost materials. The second objective was to compare biochemistry analysis results obtained from equine and southern white rhinoceros serum separated by traditional laboratory and manual salad spinner centrifugation. METHODS: We optimized the design and serum separation protocol using non-anticoagulated equine blood. Equine and rhinoceros serum samples were separated by manual salad spinner or traditional laboratory centrifugation. Measured analytes included sodium, potassium, chloride, urea nitrogen, creatinine, phosphorous, total calcium, magnesium, glucose, total protein, albumin, globulin, creatinine kinase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, total bilirubin, bicarbonate, sorbitol dehydrogenase, and triglycerides. Results obtained from serum separated by each centrifugation technique were compared by Deming regression and Bland-Altman analyses. RESULTS: A tube adaptor insert modeled after a swing angle rotor and a two-step salad spinner centrifugation yielded serum comparable to traditional laboratory centrifugation. For the majority of analytes, no proportional or constant biases were detected between centrifugation methods. A positive proportional bias in the measurement of ALP in serum separated by manual centrifugation was detected in both equine and rhinoceros samples. CONCLUSIONS: Manual centrifugation with a modified salad spinner yields diagnostic quality serum suitable for the measurement of most standard biochemistry analytes.

Ingestion of vegetable salads before rice inhibits the increase in postprandial serum glucose levels in healthy subjects.

This study aims to confirm the "vegetable-first" effect. In addition, because we aimed dietary fiber in vegetable salad, the effect after the ingestion of vegetable salad extract (vegetable salad from which solids have been removed) before carbohydrates on postprandial serum glucose level was also evaluated. A total of 13 healthy men were given meals after one-night of fasting: rice-vegetable salad, vegetable salad-rice, and vegetable salad extract-rice. Blood samples were taken at 0, 30, 45, 60, 90, and 120 min after the ingestion of the test meal to measure serum glucose levels. Serum glucose levels were significantly lower after 45 and 60 min in the vegetable salad-rice group compared to the rice-vegetable salad group. No significant difference was found between the vegetable salad extract-rice group and the vegetable salad-rice/rice-vegetable salad group. The result suggested that it might be important to ingest vegetables to obtain the "vegetable-first" effect.

Effect of high-pressure processing on the inactivation of foodborne pathogens in egg salad.

The objectives of this study were to determine the effect of high-pressure processing (HPP) on the survival of Listeria monocytogenes, Salmonella serotype Typhimurium, and Escherichia coli O157:H7 in egg salad and to evaluate the number of sub-lethally injured cells based on treatment conditions. HPP at 500 MPa for 30 s was sufficient for the complete inactivation of L. monocytogenes and Salm. Typhimurium directly plated on selective agar or plated after resuscitation, while 2 min treatment was required for E. coli O157:H7. HPP at 600 MPa for 30 s completely inactivated L. monocytogenes and Salm. Typhimurium, while 1 min treatment was needed for E. coli O157:H7. HPP at 400‒500 MPa injured a large number of pathogenic bacteria. No significant changes (P  > 0.05) in pH and color of egg salad were observed between HPP-treated and non-treated samples during 28 days of storage at refrigerated temperature. Our findings could be useful in predicting the HPP-mediated inactivation patterns of foodborne pathogens in egg salad for practical applications.

Evaluation of Covalent Organic Frameworks for the low-cost, rapid detection of Shiga Toxin-producing Escherichia coli in ready-to-eat salads.

BACKGROUND: Ready-to-eat products, such as leafy greens, must be carefully controlled as they are directly consumed without any treatment to reduce the presence of potential pathogens. Food industries, especially those that process products with short shelf-life, demand rapid detection of foodborne pathogens such as Shiga Toxin-producing Escherichia coli (STEC). In this sense, molecular methods can fulfill both requirements of turnaround time and consumer safety. The most popular rapid methods are those based on real-time PCR (qPCR) however, vegetables contain inhibitory compounds that may inhibit the amplification reaction thus, there is a need for novel sample preparation protocols. RESULTS: In the current study, a low-cost sample treatment based on sequential filtration steps was developed. This protocol was combined with covalent organic frameworks (COFs), and compared against a chelating resin, to evaluate their performance by multiplex qPCR targeting the major virulence genes of STEC, namely stx1, stx2, and eae, along with the rfbE for the specific identification of serogroup O157 due to its particularly high incidence, and an Internal Amplification Control to assess reaction inhibition. The optimized sample treatment effectively removed vegetable qPCR inhibitory compounds, and it was possible to detect STEC in spiked ready-to-eat salad samples in one working day, roughly 5 h, with an LOD50 of 8.7 CFU/25 g with high diagnostic sensitivity and specificity. The method was also assessed in samples with cold-stressed bacteria with good results, further demonstrating its applicability. SIGNIFICANCE: It was demonstrated for the first time that COFs are suitable for DNA extraction and purification. In addition to this, due to the tunable nature of these materials, it is envisioned that future modifications in terms of pore size or combination with magnetic materials, will allow to further improve their performance. In addition to this, the rapid and low-cost sample treatment protocol developed demonstrated suitable for the rapid screening of STEC vegetable samples.

A worldwide review of illness outbreaks involving mixed salads/dressings and factors influencing product safety and shelf life.

The trends toward healthy living, vegetarianism, and busy schedules have increased salad popularity. Salads are usually consumed raw without any thermal treatment, and therefore, without proper care they can become major vehicles for foodborne illness outbreaks. This review examines the microbial quality of 'dressed' salads which contain two or more vegetables/fruits and salad dressings. The possible sources of ingredient contamination, recorded illnesses/outbreaks, and overall microbial quality observed worldwide, besides the antimicrobial treatments available are discussed in detail. Noroviruses were most frequently implicated in outbreaks. Salad dressings usually play a positive role in influencing microbial quality. However, this depends on several factors like the type of contaminating microorganism, storage temperature, dressing pH and ingredients, plus the type of salad vegetable. Very limited literature exists on antimicrobial treatments that can be used successfully with salad dressings and 'dressed' salads. The challenge with antimicrobial treatments is to find ones sufficiently broad in spectrum, compatible with produce flavour which can be applied at competitive cost. It is evident that renewed emphasis on prevention of produce contamination at the producer, processor, wholesale and retail levels plus enhanced hygiene vigilance at foodservice will have a major impact on reducing the risk of foodborne illnesses from salads.

Persistence and survival of Cryptosporidium parvum oocysts on lamb's lettuce leaves during plant growth and in washing conditions of minimally-processed salads.

Cryptosporidium is the causative agent of cryptosporidiosis, which results, among others, in profuse diarrhoea. Transmission to humans occurs via the faecal-oral route directly by contact with infected hosts or indirectly by waterborne or foodborne routes. For the latter, parasite transmission is closely linked to the oocyst's ability to persist and survive in food matrices. In this study, we evaluated the persistence and survival of Cryptosporidium oocysts in lamb's lettuce: i) during plant growth and ii) in conditions mimicking the industrial washing process applied in minimally-processed vegetables (MPV). Results show that oocysts persisted during the growth of lamb's lettuce, i.e. two months from the 2-leaf stage until the 8-leaf harvest time (-0.89 Log10 of oocysts). However, their survival decreased from as early as one week (-0.61 Log10), and only 6 % of oocysts remained infective at the time of harvest. The washing process had a limited effect on parasite load (<0.5 Log10) and no effect on survival; chlorination of washing water did not improve the efficiency (removal and inactivation) of the process. The ability of C. parvum to persist and survive throughout the food chain may drive its transmission to humans through MPV products. Appropriate management measures should be implemented at each operational level to limit contamination and ensure food safety of fresh produce.

"It's about What You've Assigned to the Salad": Focus Group Discussions on the Relationship between Food and Mood.

Current observational and interventional studies in nutritional psychiatry suggest that healthy dietary patterns rich in fresh whole foods could protect against depressive symptoms, and that unhealthy dietary patterns high in ultra-processed and refined foods could contribute to depressive symptoms. However, no studies have explored detailed subjective accounts behind the food and mood relationship. This study aimed to uncover unknown factors in the human experience with food and mood. Using a phenomenological framework, this focus group study applied thematic template analysis to accounts of over 50 Australians aged between 18 and 72. Three themes were identified from the transcript of the focus groups: (i) reactive and proactive relationships with food, (ii) acknowledgement of individual diversity relating to eating and mental health, and (iii) improving mood by removing food restriction and eating intuitively. The data highlights the complexity of the relationship between food and mood that extends beyond biological mechanisms which could be used to extend current epidemiological and intervention studies in the field of dietary patterns and depression.

Ready-To-Eat Rocket Salads as Potential Reservoir of Bacteria for the Human Microbiome.

Reportedly, Western-type diets may induce the loss of key microbial taxa within the gastrointestinal microbiota, promoting the onset of noncommunicable diseases. It was hypothesized that the consumption of raw vegetables could contribute to the maintenance of the intestinal microbial community structure. In this context, we explored bacteria associated with commercial rocket salads produced through different farming practices: traditional (conventional, organic, and integrated) and vertical farming. Viable counts of mesophilic bacteria and lactic acid bacteria (LAB) were performed on plate count agar (PCA) and de Man-Rogosa-Sharpe (MRS) agar at pH 5.7, whereas metataxonomics through 16S rRNA gene sequencing was used to profile total bacteria associated with rocket salads. We found that rocket salads from vertical farming had much fewer viable bacteria and had a bacterial community structure markedly different from that of rocket salads from traditional farming. Furthermore, although α- and ß-diversity analyses did not differentiate rocket samples according to farming techniques, several bacterial taxa distinguished organic and integrated from conventional farming salads, suggesting that farming practices could affect the taxonomic composition of rocket bacterial communities. LAB were isolated from only traditional farming samples and belonged to different species, which were variably distributed among samples and could be partly associated with farming practices. Finally, the INFOGEST protocol for in vitro simulation of gastrointestinal digestion revealed that several taxonomically different rocket-associated bacteria (particularly LAB) could survive gastrointestinal transit. This study suggests that commercial ready-to-eat rocket salads harbor live bacteria that possess the ability to survive gastrointestinal transit, potentially contributing to the taxonomic structure of the human gut microbiota. IMPORTANCE Western-type diets are composed of foods with a reduced amount of naturally occurring microorganisms. It was hypothesized that a microbe-depleted diet can favor the alteration of the human intestinal microbial ecosystem, therefore contributing to the onset of chronic metabolic and immune diseases currently recognized as the most significant causes of death in the developed world. Here, we studied the microorganisms that are associated with commercial ready-to-eat rocket salads produced through different farming practices. We showed that rocket salad (a widely consumed vegetal food frequently eaten raw) may be a source of lactic acid bacteria and other microbes that can survive gastrointestinal transit, potentially increasing the biodiversity of the intestinal microbiota. This deduction may be valid for virtually all vegetal foods that are consumed raw.

Effect of cold atmospheric plasma torch distance on the microbial inactivation and sensorial properties of ready-to-eat olivier salad.

This study aimed to investigate the effect of the cold atmospheric plasma torch (CAPT) nozzle distance from the surface of Olivier salad and the treatment time in the reduction of microbial load and sensory properties of the product simultaneously. In this study, the CAPT nozzle was placed at 3, 5, and 7 cm distances from the surface of the Olivier salad, and its efficiency in inactivating the microbial population, decimal reduction time (D-value), and sensory evaluation of the product were evaluated. The results showed that reducing the distance and increasing the plasma treatment time (30, 60, 90, and 120 s) both reduced the microbial load of the product. The maximum inactivation and the minimum D-value are related to the 3 cm distance for 120 s, which has been 3.77, 2.91, and 1.52 log CFU/g for Coliform, Total viable count (TVC), mold and yeast, respectively. The lowest D-value was related to Coliform (4.41 s). CAPT treatment had no significant sensible effect on the product's sensory characteristics compared to the control sample. The treated sample at a 3 cm distance for 90 s and the microbial reduction to an acceptable amount and high acceptancy from sensory evaluators were selected as the superior treatment in this study. Also, the results showed that CAPT could be used successfully in ready-to-eat (RTE) products.