Serotype Enteritidis Research Articles

Whole genome sequencing and protein structure analyses of target genes for the detection of Salmonella

Rapid and sensitive detection of Salmonella is a critical step in routine food quality control, outbreak investigation, and food recalls. Although various genes have been the targets in the design of rapid molecular detection methods for Salmonella, there is limited information on the diversity of these target genes at the level of DNA sequence and the encoded protein structures. In this study, we investigated the diversity of ten target genes (invA, fimA, phoP, spvC, and agfA; ttrRSBCA operon including 5 genes) commonly used in the detection and identification of Salmonella. To this end, we performed whole genome sequencing of 143 isolates of Salmonella serotypes (Enteritidis, Typhimurium, and Heidelberg) obtained from poultry (eggs and chicken). Phylogenetic analysis showed that Salmonella ser. Typhimurium was more diverse than either Enteritidis or Heidelberg. Forty-five non-synonymous mutations were identified in the target genes from the 143 isolates, with the two most common mutations as T ↔ C (15 times) and A ↔ G (13 times). The gene spvC was primarily present in Salmonella ser. Enteritidis isolates and absent from Heidelberg isolates, whereas ttrR was more conserved (0 non-synonymous mutations) than ttrS, ttrB, ttrC, and ttrA (7, 2, 2, and 7 non-synonymous mutations, respectively). Notably, we found one non-synonymous mutation (fimA-Mut.6) across all Salmonella ser. Enteritidis and Salmonella ser. Heidelberg, C → T (496 nt postion), resulting in the change at AA 166 position, Glutamine (Q) → Stop condon (TAG), suggesting that the fimA gene has questionable sites as a target for detection. Using Phyre2 and SWISS-MODEL software, we predicted the structures of the proteins encoded by some of the target genes, illustrating the positions of these non-synonymous mutations that mainly located on the α-helix and β-sheet which are key elements for maintaining the conformation of proteins. These results will facilitate the development of sensitive molecular detection methods for Salmonella.

Nontyphoidal Salmonella in Food from Latin America: A Systematic Review.

In Latin America, nontyphoidal Salmonella (NTS) is one of the most important etiological agents of foodborne infections; it can survive in soil, water, and food even after processing. Here, we aimed to perform a systematic review by collecting data on the prevalence, serotypes, and antimicrobial resistance (AMR) of NTS isolated from different food products in Latin America, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Out of 1766 studies screened, 244 reports from 13 Latin American countries were eligible. Among these, 182 reported NTS prevalence, 87 reported NTS serotypes, and 83 reported serotypes with AMR patterns. The NTS prevalence ranged from 0.005% to 93.3%, regardless of country and food. Meat showed the highest NTS prevalence. Enteritidis, Typhimurium, and Derby were the most frequently observed serotypes in different food products. The serotypes Enteritidis, Typhimurium, and Infantis, isolated from animal products, showed the highest AMR rate. The presence of NTS in fruits and vegetables, which are generally consumed raw or as ready-to-eat food, indicates a high risk of salmonellosis from consuming these foods. Thus, the reduction of this pathogen in the food chain requires a One Health approach, involving good agricultural and manufacturing practices, low antimicrobial use, and proper waste management.

Genomic Characterization of Endemic and Ecdemic Non-typhoidal Salmonella enterica Lineages Circulating Among Animals and Animal Products in South Africa.

In Africa, the burden of illness caused by non-typhoidal Salmonella enterica is disproportionally high; however, whole-genome sequencing (WGS) efforts are overwhelmingly concentrated in world regions with lower burdens. While WGS is being increasingly employed in South Africa to characterize Salmonella enterica, the bulk of these efforts have centered on characterizing human clinical strains. Thus, very little is known about lineages circulating among animals in the country on a genomic scale. Here, we used WGS to characterize 63 Salmonella enterica strains isolated from livestock, companion animals, wildlife, and animal products in South Africa over a 60-year period. Genomes were assigned to serotypes Dublin, Hadar, Enteritidis, and Typhimurium (n = 18, 8, 13, and 24 strains, respectively) and sequence types (STs) ST10 (all S. Dublin), ST33 (all S. Hadar), ST11/ST366 (n = 12 and 1 S. Enteritidis, respectively), and ST19/ST34 (n = 23 and 1 S. Typhimurium, respectively; via seven-gene multi-locus sequence typing). Within-ST phylogenies were constructed using genomes sequenced in this study, plus publicly available genomes representative of each ST’s (i) global (n = 2,802 and 1,569 S. Dublin and Hadar genomes, respectively) and (ii) African (n = 716 and 343 S. Enteritidis and Typhimurium genomes, respectively) population. For S. Dublin ST10, a largely antimicrobial-susceptible, endemic lineage circulating among humans, animals, and food in South Africa was identified, as well as a lineage that was likely recently introduced from the United States. For S. Hadar ST33, multiple South African lineages harboring streptomycin and tetracycline resistance-conferring genes were identified. African S. Enteritidis ST11 could be primarily partitioned into one largely antimicrobial-susceptible and one largely multidrug-resistant (MDR) clade, with South African isolates confined to the largely antimicrobial-susceptible clade. S. Typhimurium ST19/ST34 strains sequenced here were distributed across the African S. Typhimurium ST19/ST34 phylogeny, representing a diverse range of lineages, including numerous MDR lineages. Overall, this study provides critical insights into endemic and ecdemic non-typhoidal Salmonella enterica lineages circulating among animals, foods, and humans in South Africa and showcases the utility of WGS in characterizing animal-associated strains from a world region with a high salmonellosis burden.

Immunization of chickens with Salmonella gallinarium ghosts expressing Salmonella Enteritidis NFliC-FimAC and CD40LC fusion antigen enhances cell-mediated immune responses and protects against wild-type challenges with both species

This study describes the construction and immunological characterization of a novel Salmonella gallinarium ghost vaccine to protect against S. gallinarium (SG) and S. Enteritidis (SE) serotypes. The SG ghost was designed to express N-terminus FliC (D0-D1 domain) and FimA retrieved from the SE genome, and the receptor-binding domain (RBD) of CD40L from the chicken as a single fusion construct. The construct was built in pJHL184, a phage lysis gene E-mediated ghost plasmid and the expression was confirmed by western blot resulting in an 85-kDa band. Chicken immunization was conducted by intramuscular route with SG ghost FliC-FimA-CD40L, vector control, or PBS alone in a prime-boost schedule. Antibody responses, cell-mediated immune responses (CMI), and cytokine induction was assessed in chicken demonstrating significantly high levels of IgY, CMI, cytokine responses in ghost immunized group delivering partial protection against SG wild type challenge and near complete protection against SE challenge wild type challenge.

Lateral flow strip biosensor based on streptavidin-coated gold nanoparticles with recombinase polymerase amplification for the quantitative point-of-care testing of Salmonella

Salmonella is a widespread and highly infectious biological contaminant that causes zoonotic diseases. Most detection methods are qualitative, while quantitative methods are time-consuming and rely on large analytical instruments. Therefore, the development of a rapid, simple, and quantitative assay is important for the prevention and control of infection at the early stages of an outbreak. To address this requirement, we developed a lateral flow (LF) strip biosensor based on streptavidin-coated gold nanoparticles (StreptAv–AuNPs) combined with recombinase polymerase amplification (RPA) for the quantitative point-of-care testing of Salmonella enterica serotype Enteritidis (S. Enteritidis). StreptAv-AuNPs were simple to synthesise and bound firmly to DNA with end-modified biotin. RPA could greatly reduce detection time. A smartphone was used to collect images of the LF strip and input them into a laptop for quantitative analysis. The entire process was completed in less than 40 min and did not require large laboratory equipment or analytical instruments. A limit of detection of 5.5 × 10−13 M and 91.4 CFU/mL was achieved for synthetic S. Enteritidis DNA and cultured S. Enteritidis samples, respectively. In the reproducibility assay, RSD values of 5.88%, 5.13%, and 4.45% were achieved for the three different concentrations of samples. This method has the advantages of low cost, high precision and rapid quantification, it is also suitable for the detection of various DNA-containing analytes.

Global spread of Salmonella Enteritidis via centralized sourcing and international trade of poultry breeding stocks

A pandemic of Salmonella enterica serotype Enteritidis emerged in the 1980s due to contaminated poultry products. How Salmonella Enteritidis rapidly swept through continents remains a historical puzzle as the pathogen continues to cause outbreaks and poultry supply becomes globalized. We hypothesize that international trade of infected breeding stocks causes global spread of the pathogen. By integrating over 30,000 Salmonella Enteritidis genomes from 98 countries during 1949–2020 and international trade of live poultry from the 1980s to the late 2010s, we present multifaceted evidence that converges on a high likelihood, global scale, and extended protraction of Salmonella Enteritidis dissemination via centralized sourcing and international trade of breeding stocks. We discovered recent, genetically near-identical isolates from domestically raised poultry in North and South America. We obtained phylodynamic characteristics of global Salmonella Enteritidis populations that lend spatiotemporal support for its dispersal from centralized origins during the pandemic. We identified concordant patterns of international trade of breeding stocks and quantitatively established a driving role of the trade in the geographic dispersal of Salmonella Enteritidis, suggesting that the centralized origins were infected breeding stocks. Here we demonstrate the value of integrative and hypothesis-driven data mining in unravelling otherwise difficult-to-probe pathogen dissemination from hidden origins.

A detailed analysis of innate and adaptive immune responsiveness upon infection with Salmonella enterica serotype Enteritidis in young broiler chickens

Salmonella enterica serotype Enteritidis (SE) is a zoonotic pathogen which causes foodborne diseases in humans as well as severe disease symptoms in young chickens. More insight in innate and adaptive immune responses of chickens to SE infection is needed to understand elimination of SE. Seven-day-old broiler chickens were experimentally challenged with SE and numbers and responsiveness of innate and adaptive immune cells as well as antibody titers were assessed. SE was observed in the ileum and spleen of SE-infected chickens at 7 days post-infection (dpi). At 1 dpi numbers of intraepithelial cytotoxic CD8+ T cells were significantly increased alongside numerically increased intraepithelial IL-2Rα+ and 20E5+ natural killer (NK) cells at 1 and 3 dpi. At both time points, activation of intraepithelial and splenic NK cells was significantly enhanced. At 7 dpi in the spleen, presence of macrophages and expression of activation markers on dendritic cells were significantly increased. At 21 dpi, SE-induced proliferation of splenic CD4+ and CD8+ T cells was observed and SE-specific antibodies were detected in sera of all SE-infected chickens. In conclusion, SE results in enhanced numbers and activation of innate cells and we hypothesized that in concert with subsequent specific T cell and antibody responses, reduction of SE is achieved. A better understanding of innate and adaptive immune responses important in the elimination of SE will aid in developing immune-modulation strategies, which may increase resistance to SE in young broiler chickens.

Impacts of Microbial Food Safety in China and Beyond.

Impacts of Microbial Food Safety in China and Beyond.

The application of a novel non-thermal plasma device with double rotary plasma jets for inactivation of Salmonella Enteritidis on shell eggs and its effects on sensory properties

Consumer awareness and distaste towards both bacterial and chemical contaminations on food items have been increasing in recent years. Non-thermal plasma (NTP) is a cutting-edge technology which has been shown to effectively inactivate bacteria on the treated foods. Although the general NTP with a single plasma jet is appropriate for the continuous operation process, it suffers limitations due to its smaller scanning area. Here, a novel NTP device with a double rotary nozzle jet system was utilized, which could treat an area instead of a point. The shell eggs inoculated with Salmonella enterica serotype Enteritidis (SE) were placed on a moving platform under the double rotary nozzle jet system. The efficacy of the NTP treatment on microbial decontamination was evaluated by testing a total of 26 combinations of operating parameters consisting of various plasma power (150, 180, 210 W), argon flow rate (10, 15, 20 slm), repetition of the moving platform (4, 6, 8 times), and speed of the moving platform (5, 10 mm/s). Although significantly higher SE reduction (p < 0.05) was achieved with higher power, more repetitions, larger argon flow rates, and lower speed of the platform, these parameters induced significant alterations in the sensory properties of the treated eggs. By comprehensively considering the bacterial reductions, egg quality, and sensory properties, NTP treatment with combination T (180 W-15 slm-6 times-10 mm/s) was determined to be the optimal parameter, which achieved >4 log CFU/egg of SE reduction and significantly better sensory properties than commercially washed eggs (p < 0.05). Additionally, SEM analysis revealed that NTP treatment with combination T resulted in less damage to egg cuticles compared to commercially washed eggs. This novel NTP device offers an efficient antibacterial activity under shorter exposure time (30 s), smaller argon flow rate (15 slm), and lower power (180 W) without adversely affecting the overall quality of the treated eggs. Therefore, this NTP device equipped with the double rotary jet system possesses a potential solution for future industrial applications.

Antimicrobial resistance patterns, virulence gene profiles, and genetic diversity of Salmonella enterica serotype Enteritidis isolated from patients with gastroenteritis in various Iranian cities.

Objective(s):This study aimed to evaluate antibiotic resistance profiles and presence of virulence genes among Salmonella enterica serovar Enteritidis (S. Enteritidis) isolated from patients with gastroenteritis in various regions of Iran. Moreover, genetic relatedness among the strains was assessed by pulsed-field gel electrophoresis (PFGE).Materials and Methods:From April through September 2017, 59 Salmonella strains were isolated from 2116 stool samples. Of these strains, 27 S. Enteritidis were recovered. These strains were subjected to disk diffusion tests, polymerase chain reaction (PCR) for detection of virulence genes (invA, hilA, pefA, rck, stn, ssrA, ssaR, sefA, spvC, sipA, sipC, sopB, sopE, and sopE2), and PFGE.Results:High prevalence of resistance towards cefuroxime (n = 20, 74.1%) and ciprofloxacin (n = 13, 48.2%) were demonstrated. All tested strains possessed invA, hilA, sefA, sipA, sopB, and sopE. The least prevalent virulence gene was rck (n = 6; 22.2%). Based on combinations of virulence genes, 12 virulotypes were observed. The most common virulotype was VP2 (n = 12; 44.4%), harboring all of the virulence genes except for rck. PFGE typing showed only two distinct fingerprints among tested strains. Each fingerprint had completely different virulotypes. Notably, VP4 (harboring all genes except for rck and spvC) was only presented in pulsotype A, while VP2 was confined to pulsotype B.Conclusion: S. Enteritidis strains were derived from a limited number of clones, suggesting that it is highly homogenous. Future works should consider combinations of other genotyping methods together with larger sample sizes from more diverse sources.

Evaluation of Pediococcus pentosaceus strains as probiotic adjunct cultures for soybean milk post-fermentation

Soybean milk is an economical substitute for dairy products. Pediococcus pentosaceus has been used as a food additive to improve taste, nutrition, and food safety. In this study, four P. pentosaceus strains (CICC 24444, QK-1, MQ-1 and RQ-1) isolated from various food sources and known to exhibit broad-spectrum antibacterial activities were used to ferment soybean milk, and their fermentation characteristics and the properties of the resulting beverages were evaluated. The results revealed that the P. pentosaceus strains can inhibited the growth of five types of pathogenic bacteria (Salmonella enterica subsp. enterica serotype Enteritidis, Yersinia enterocolitica, Shigella dysenteriae, Escherichia coli, and Staphylococcus aureus), and their in vitro survival rates in the simulated stomach and intestinal environments were above 90%, satisfying the probiotic requirements. Isomaltose oligosaccharide was used as a protective agent to resist low-temperature freeze-drying damage and ensure a high survival rate, and P. pentosaceus was directly injected into fermented soymilk. The acidification of fermented soybean milk was the weakest with P. pentosaceus QK-1, and the viable bacterial counts of all strains were stable after 28 days of storage. After fermentation, the antioxidant ability was enhanced. Arginine and β-alanine levels increased after fermentation, and the adjunct culture of P. pentosaceus QK-1 increased proline levels. Our data indicate that P. pentosaceus QK-1 is a suitable strain for the development of functional plant-based beverages.

Contamination of street food with multidrug-resistant Salmonella, in Ouagadougou, Burkina Faso.

BackgroundGastrointestinal infections are a global public health problem. In Burkina Faso, West Africa, exposure to Salmonella through the consumption of unhygienic street food represents a major risk of infection requiring detailed evaluation.MethodsBetween June 2017 and July 2018, we sampled 201 street food stalls, in 11 geographic sectors of Ouagadougou, Burkina Faso. We checked for Salmonella contamination in 201 sandwiches (one per seller), according to the ISO 6579:2002 standard. All Salmonella isolates were characterized by serotyping and antimicrobial susceptibility testing, and whole-genome sequencing was performed on a subset of isolates, to investigate their phylogenetic relationships and antimicrobial resistance determinants.ResultsThe prevalence of Salmonella enterica was 17.9% (36/201) and the Salmonella isolates belonged to 16 different serotypes, the most frequent being Kentucky, Derby and Tennessee, with five isolates each. Six Salmonella isolates from serotypes Brancaster and Kentucky were multidrug-resistant (MDR). Whole-genome sequencing revealed that four of these MDR isolates belonged to the emergent S. enterica serotype Kentucky clone ST198-X1 and to an invasive lineage of S. enterica serotype Enteritidis (West African clade).ConclusionThis study reveals a high prevalence of Salmonella spp. in sandwiches sold in Ouagadougou. The presence of MDR Salmonella in food on sale detected in this study is also matter of concern.

The Formation of Two Hybrid Plasmids Mediated by IS26 and Tn6952 in Salmonella enterica Serotype Enteritidis.

To characterize the formation mechanism and characteristics of two cointegrate plasmids in Salmonella enterica serotype Enteritidis strain S13, plasmids from strain S13 and three corresponding transconjugants were subjected to whole genome sequencing and analyzed using bioinformatics tools. The traits of two fusion plasmids in transconjugants were characterized by stability and conjugation experiments. Sequence analysis indicated that strain S13 contained four plasmids, including mcr-1-bearing pS13-1, blaCTX–M–55-carrying pS13-2, tet(M)-bearing pS13-3, and floR-carrying pS13-4. IncN1-F33:A–:B– plasmid pS13-2, respectively, fused with IncFI:A–:B– plasmid pS13-3 and IncX1 plasmid pS13-4, which generated two cointegrate plasmids, designated pS13D and pS13F, which involved in two intermolecular replicative mechanisms mediated by IS26 and the novel transposon Tn6952 (ΔTnAS3-IS26-ΔISEcp1-ramA-ΔIS26-ΔTnAS1), respectively. This is the first report of the fusion of the IncN1-F33:A–:B– plasmid and IncFI:A–:B– plasmid mediated by IS26, and with IncX1 plasmid mediated by Tn6952. The formation and evolution of cointegrate plasmids could expand the resistance and host spectrum of fusion plasmids.

Characterization of Salmonella Isolates Recovered from Stages of the Processing Lines at Four Broiler Processing Plants in Trinidad and Tobago.

This cross-sectional study determined the prevalence, characteristics, and risk factors for contamination of chicken with Salmonella at four operating broiler processing plants in Trinidad. Standard methods were used to isolate and characterize the Salmonella isolates. The overall prevalence of Salmonella at the four processing plants was 27.0% (107/396). The whole carcass enrichment (WCE) method yielded a statistically significantly (p = 0.0014) higher frequency of isolation (53.9%; 97/180) than the whole carcass rinse (35.0%; 63/180) and neck skin methods (42.2%; 38/90). S. enterica serotypes Enteritidis, Javiana, and Infantis were the predominant serotypes isolated accounting for 20.8%, 16.7% and 12.5%, respectively, of the serotyped isolates. Risk factors included the use of over 100 contract farmers (OR 4.4), pre-chiller (OR 2.3), addition of chlorine to chiller (OR 3.2), slaughtering sick broilers (OR 4.4), and flocks with >50% mortality. Multi-drug resistance was detected in 12.3% (14/114) of the isolates of Salmonella. Resistance was high to kanamycin (85.7%) and doxycycline (74.6%) but low to amoxicillin-clavulanic acid (2.4%) and sulphamethoxazole-trimethoprim (0.8%). The occurrence of resistant Salmonella in chickens processed at commercial broiler processing plants has implications for salmonellosis and therapeutic failure in consumers of improperly cooked contaminated chickens from these plants in the country.

Molecular characterisation of antimicrobial resistance determinants and class 1 integrons of Salmonella enterica subsp. enterica serotype Enteritidis strains from retail food in China

Salmonella is a genus responsible for a majority of foodborne disease outbreaks affecting public health worldwide, and Salmonella enterica subsp. enterica serotype Enteritidis (S. Enteritidis) is the most common pathogen that presents alarming rates of antimicrobial resistance. The aim of this study was to analyse the antimicrobial resistance of 124 S. Enteritidis strains collected from retail foods in 39 cities across China and to conduct a comprehensive molecular characterisation of their antimicrobial resistance genes (ARGs) and class 1 integrons through PCR amplification, antibiotic susceptibility testing, and conjugation assays. The strains exhibited a multidrug resistance rate as high as 68.5% and were frequently resistant to nalidixic acid (97.6%) and ampicillin (74.2%). Correspondingly, the analysis of quinolone genes revealed that 97.5% of the strains harboured single mutations in gyrA; some possessed the plasmid-mediated quinolone resistance genes qnrS (0.8%), oqxAB (2.4%), as well as the blaTEM-1 (67.7%); and the extended spectrum beta-lactamase (ESBL) genes blaCTX-M-55 were detected in 2.4% of the strains. Class 1 integrons were observed in 11.3% (14/124) of the strains and integrated three different gene cassettes, namely, aadA22 (1/14), dfrA1-aadA1 (5/14), and dfrA17-aadA5 (8/14). Conjugation experiments and antimicrobial susceptibility testing showed that the integrons including antimicrobial resistance genes could be transferred to the recipient strain via mobile plasmids, rendering the recipient strain resistant. Both multilocus variable-number tandem-repeat analysis (MLVA) and core genome multilocus sequence typing (cgMLST) show a better discrimination in this study, hence a combination of MLVA and cgMLST analyses would improve the accuracy and cost-effectiveness. This study provides a systematic and comprehensive analysis of the antimicrobial resistance profiles of S. Enteritidis strains isolated from retail food in China. The present data will help to gain insight into the public health impact of S. Enteritidis and guide the development of control strategies.

Molecular Epidemiology of Salmonellosis in Florida, USA, 2017-2018.

The state of Florida reports a high burden of non-typhoidal Salmonella enterica with approximately two times higher than the national incidence. We retrospectively analyzed the population structure and molecular epidemiology of 1,709 clinical isolates from 2017 and 2018. We found 115 different serotypes. Rarefaction suggested that the serotype richness did not differ between children under 2 years of age and older children and adults and, there are ~22 well-characterized dominant serotypes. There were distinct differences in dominant serotypes between Florida and the USA as a whole, even though S. Enteritidis and S. Newport were the dominant serotypes in Florida and nationally. S. Javiana, S. Sandiego, and S. IV 50:z4, z23:- occurred more frequently in Florida than nationally. Legacy Multi Locus Sequence Typing (MLST) was of limited use for differentiating clinical Salmonella isolates beyond the serotype level. We utilized core genome MLST (cgMLST) hierarchical clusters (HC) to identify potential outbreaks and compared them to outbreaks detected by Pulse Field Gel Electrophoresis (PFGE) surveillance for five dominant serotypes (Enteritidis, Newport, Javiana, Typhimurium, and Bareilly). Single nucleotide polymorphism (SNP) phylogenetic-analysis of cgMLST HC at allelic distance 5 or less (HC5) corroborated PFGE detected outbreaks and generated well-segregated SNP distance-based clades for all studied serotypes. We propose “combination approach” comprising “HC5 clustering,” as efficient tool to trigger Salmonella outbreak investigations, and “SNP-based analysis,” for higher resolution phylogeny to confirm an outbreak. We also applied this approach to identify case clusters, more distant in time and place than traditional outbreaks but may have been infected from a common source, comparing 176 Florida clinical isolates and 1,341 non-clinical isolates across USA, of most prevalent serotype Enteritidis collected during 2017–2018. Several clusters of closely related isolates (0–4 SNP apart) within HC5 clusters were detected and some included isolates from poultry from different states in the US, spanning time periods over 1 year. Two SNP-clusters within the same HC5 cluster included isolates with the same multidrug-resistant profile from both humans and poultry, supporting the epidemiological link. These clusters likely reflect the vertical transmission of Salmonella clones from higher levels in the breeding pyramid to production flocks.

Deteksi Salmonella spp. pada Telur Ayam Konsumsi dari Peternakan Ayam Ras dan Pasar Tradisional di Bali

This study aims to early detect Salmonella spp. and identify serotypes in commercial chicken eggs from layer chicken farms and traditional markets in Bali. Early detection study of Salmonella spp. was carried out by conventional bacteriological methods, while serotype identification by duplex Polymerase Chain Reaction (d-PCR) test against the invA gene from Salmonella spp. and the sefA gene from Salmonella enteritidis. Egg samples in this study were taken from 10 layer chicken farms in Bali which included districts of Bangli, Gianyar, Tabanan and Karangasem. Egg samples from traditional markets were taken from 18 traditional markets from the districts of Bangli, Gianyar, Tabanan, Karangasem, Badung, and Denpasar City. Samples were eggshells and egg whites. Analysis of positive results from Salmonella spp. described descriptively. The results showed that eggshells and white eggs from all of the layer chicken farms are negative contaminated with Salmonella spp. (0%). In eggshell samples taken from the traditional markets of Taman Bali and Tulikup from the districts of Bangli and Gianyar, positive with Salmonella spp. (11,1%) by conventional bacteriological tests. In the duplex Polymerase Chain Reaction test, S. enteritidis serotypes were identified. The finding contamination of Salmonella enteritidis in commercial chicken eggs from traditional markets require periodically detection to prevent the occurrence of salmonellosis due to consumption of contaminated chicken eggs in traditional markets in Bali.

Integrated Whole-Genome Sequencing Infrastructure for Outbreak Detection and Source Tracing of Salmonella enterica Serotype Enteritidis.

As an important foodborne pathogen, Salmonella enterica serotype Enteritidis is recognized as one of the most common causes of human salmonellosis globally. Outbreak detection for this highly homogenous serotype, however, has remained challenging. Rapid advances in sequencing technologies have presented whole-genome sequencing (WGS) as a significant advancement for source tracing and molecular typing of foodborne pathogens. A retrospective analysis was conducted using Salmonella Enteritidis isolates (n = 65) from 11 epidemiologically confirmed outbreaks and a collection of contemporaneous sporadic isolates (n = 258) during 2007-2017 to evaluate the performance of WGS in delineating outbreak-associated isolates. Whole-genome single-nucleotide polymorphism (SNP)-based phylogenetic analysis revealed well-supported clades in concordance with epidemiological evidence and pairwise distances of ≤3 SNPs for all outbreaks. WGS-based framework of outbreak detection was thus proposed and applied prospectively to investigate isolates (n = 66) from nine outbreaks during 2018-2019. We further demonstrated the superior discriminatory power and accuracy of WGS to resolve and delineate outbreaks for pragmatic food source tracing. The proposed integrated WGS framework is the first in China for Salmonella Enteritidis and has the potential to serve as a paradigm for outbreak detection and source tracing of Salmonella throughout the stages of food production, as well as expanded to other foodborne pathogens.

Spray and Aerosolised pH-Neutral Electrochemically Activated Solution Reduces Salmonella Enteritidis and Total Bacterial Load on Egg Surface

The effectiveness of sprayed and aerosolised pH-neutral electrochemically activated solutions (ECAS) containing 150 mg/L of free available chlorine in reducing total bacteria load and artificially inoculated Salmonella enterica serotype Enteritidis 11RX on eggs surfaces was investigated. Treatment groups included untreated control, sodium hypochlorite (positive control), sprayed and aerosolised water and sprayed and aerosolised ECAS. Sprayed ECAS (150 mg/L, 45 s) showed a significant reduction in total bacterial load (2.2 log reduction, p &lt; 0.0001) and S. Enteritidis (5.4 log reduction, p &lt; 0.0001) when compared with the untreated control. Aerosolised ECAS (120 s) was effective in reducing both the total bacterial load (1.4 log reduction, p &lt; 0.01) and S. Enteritidis (4.2 log reduction, p = 0.0022). However, aerosolised ECAS (60 s) only significantly reduced S. Enteritidis counts (2.8 log reduction, p &lt; 0.0008), indicating that a longer time for bacterial reduction during fogging sanitisation is needed. Tests performed with one egg per oscillating tray were more effective in reducing both the total bacterial load and the S. Enteritidis counts than those with three eggs per oscillating tray. Sprayed ECAS (45 s) and aerosolised ECAS (120 s) did not deteriorate the egg cuticle integrity (ΔEab*), which was evaluated using Cuticle Blue dye solution and colour intensity measurement. Overall, both the reduction in total bacteria counts and S. Enteritidis from the egg surface and retention of cuticle integrity suggest that sprayed and aerosolised ECAS could be used as alternative sanitising approaches to improve the food safety aspect of table eggs.

Whole-Genome Sequencing Analysis of Salmonella Enterica Serotype Enteritidis Isolated from Poultry Sources in South Korea, 2010-2017.

Salmonella enterica subsp. enterica serotype Enteritidis (SE) is recognized as a major cause of human salmonellosis worldwide, and most human salmonellosis is due to the consumption of contaminated poultry meats and poultry byproducts. Whole-genome sequencing (data were obtained from 96 SE isolates from poultry sources, including an integrated broiler supply chain, farms, slaughterhouses, chicken transporting trucks, and retail chicken meats in South Korea during 2010–2017. Antimicrobial resistance and virulence genes were investigated using WGS data, and the phylogenetic relationship of the isolates was analyzed using single-nucleotide polymorphism (SNP) typing and core genome multilocus sequence typing (cgMLST). All isolates carried aminoglycoside resistance genes, aac(6’)-Iaa, and 56 isolates carried multiple antimicrobial resistance genes. The most frequent virulence gene profile, pef-fim-sop-inv.-org-sip-spa-sif-fli-flg-hil-ssa-sse-prg-pag-spv, was found in 90 isolates. The SNP analysis provided a higher resolution than the cgMLST analysis, but the cgMLST analysis was highly congruent with the SNP analysis. The phylogenetic results suggested the presence of resident SE within the facility of processing plants, environments of slaughterhouses, and the integrated broiler supply chain, and the phylogenetically related isolates were found in retail meats. In addition, the SE isolates from different origins showed close genetic relationships indicating that these strains may have originated from a common source. This study could be valuable reference data for future traceback investigations in South Korea.