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Trans-cinnamaldehyde (TC), a typical plant-derived compound, has been widely used in the control of foodborne pathogen contamination. Nevertheless, the risk associated with the occurrence of viable but nonculturable (VBNC) bacteria induced by TC remains unclear. The results of this study showed that Salmonella Enteritidis ( S. Enteritidis) entered the VBNC state after being induced by TC at a minimum inhibitory concentration of 312.5 μg/mL and survived for at least 22 days under TC treatment. Enhanced resistance was found against heat treatment (75°C, 30 s), antibiotics (i.e., ampicillin, ceftriaxone sodium, chloramphenicol), and hydrogen peroxide (3%) in VBNC S. Enteritidis. A synergistic effect against VBNC S. Enteritidis occurred when TC was combined with acid treatment, including lactic acid and acetic acid (pH = 3.5). VBNC and resuscitated S . Enteritidis by sodium pyruvate treatment (100 mM) were found to retain the infectious ability to Caco-2 cells. Relative expression levels of the stress-related genes relA , spoT , ppx , lon , katG , sodA , dnaK , and grpE were upregulated in VBNC S . Enteritidis. Accumulation of reactive oxygen species (ROS) and protein aggregates was observed in VBNC cells. Besides, the resuscitation of VBNC cells was accompanied with clearance of ROS and protein aggregates. In summary, this study presents a comprehensive characterization of stress tolerance and resuscitation of VBNC S . Enteritidis induced by cinnamaldehyde, and the results provide useful information for the development of effective control strategy against VBNC pathogenic bacteria in food production.

In China, Salmonella is one of the most frequent causes of bacterial gastroenteritis, and food handlers in restaurants as an important contaminated source were rarely reported. In May 2023, an outbreak of Salmonella enterica serovar Enteritidis infection in a restaurant in Jiangxi Province, China, was investigated. Cases were interviewed. Stool samples from cases, anal swabs from restaurant employees, suspicious raw food materials, and semifinished food were collected and examined. Pulsed-field gel electrophoresis (PFGE) and whole genome sequencing (WGS) were performed to determine the relatedness of the pathogen isolates. Antimicrobial resistance genes and virulence genes of isolates were analyzed by WGS. The antimicrobial profile of the isolates was detected by broth microdilution, which involved 20 different antibiotics. Among the 31 patrons, 26 showed gastrointestinal symptoms. Five Salmonella Enteritidis strains were isolated from patients (2), semifinished food (2), and food handler (1). The results of PFGE and single-nucleotide polymorphism showed that these five isolates were identical clones. These findings demonstrated that this outbreak was a restaurant Salmonella Enteritidis outbreak associated with an infected food handler. The rates of resistance to nalidixic acid and colistin and intermediate resistance to ciprofloxacin were 100%, 80%, and 100%, respectively. These outbreak isolates harbored point mutation gyrA p.D87G. The cause of inconsistency between the genotype and phenotype of resistance was deeply discussed. A total of 107 virulence genes were found in each isolate, with many being associated with Salmonella pathogenicity island (SPI)-1 and SPI-2. As an overlooked contamination source, infected food handlers can easily cause large-scale outbreaks. This outbreak highlighted that the government should enhance the training and supervision of food hygiene and safety for food handlers to prevent foodborne outbreaks.

Objectives: This study aimed to trace the transmission source of Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Enteritidis strains associated with enteric infections in Shanghainese children, and understand the molecular mechanism of resistance to third-generation cephalosporins and ciprofloxacin., Materials and Methods: The profiles of pulsed-field gel electrophoresis (PFGE) were compared among the isolates from children, animal, and environment. Antimicrobial susceptibility was determined using the minimal inhibitory concentrations and Kirby-Bauer disk diffusion method. Extended-spectrum β-lactamase (ESBL) producing isolates mediated by resistance genes were identified using polymerase chain reaction and sequencing., Results: Based on PFGE patterns, 49 (33.1%) of 148 human Salmonella Typhimurium isolates located in the dominant PFGE clusters were genetically related to the isolates from poultry source, environment water, aquatic products, and reptiles, whereas 97 (97.0%) of 100 human Salmonella Enteritidis isolates were genetically related to isolates from poultry and water. The rates of resistance to ceftriaxone among clinical Salmonella Typhimurium and Salmonella Enteritidis isolates were 42.0% and 14.2%, respectively. Besides, 35.1% of Salmonella Typhimurium isolates displayed resistance to ciprofloxacin; 64.9% of Salmonella Typhimurium isolates and 97.0% of Salmonella Enteritidis isolates displayed reduced susceptibility to ciprofloxacin. Of 64 ESBL/AmpC-producing strains, CTX-M, TEM, DHA, and CMY were detected at frequencies of 86.0%, 62.5%, 7.8%, 3.1%, and 3.1%, respectively., Conclusions: The transmission sources of Salmonella Typhimurium and Salmonella Enteritidis infections in Shanghainese children were diverse. The high prevalence of resistance to third-generation cephalosporins and ciprofloxacin mediated by multiple molecular mechanisms needs continuous monitoring and intervention.

To determine the distribution of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of essential oils (EOs) of cinnamon (Cinnamomum zeylanicum), clove (Eugenia caryophyllata), oregano (Origanum vulgare), common thyme (Thymus vulgaris), and red thyme (Thymus zygis) against Salmonella enterica, double serial dilutions of each EO were challenged with 85 Salmonella strains belonging to 23 serotypes of animal origin. The results showed the bactericidal character of the EOs tested against S. enterica, highlighting the oregano with MIC 50 and MBC 50 of 3.12 × 10 -4  g/mL, and MIC 90 and MBC 90 of 6.25 × 10 -4  g/mL. When comparing the Salmonella Typhimurium and Salmonella Enteritidis serotypes susceptibility, we observed a significantly higher sensitivity of Typhimurium to clove and Enteritidis to cinnamon. In addition, Typhimurium isolates with significantly higher MIC and MBC values for all the EOs tested were found, suggesting the existence of a possible resistance profile. The results of this study provide relevant data for the potential of EOs as antibacterials, although they highlight the need to continue bacterial sensitivity distribution studies and consider the differences detected for future in vivo studies.

Salmonella enterica serovar Enteritidis is the leading global cause of salmonellosis. A total of 146 Salmonella Enteritidis isolates obtained from retail chicken products in Shanghai, China were characterized for their antimicrobial susceptibilities, virulence and antibiotic resistance gene profiles, and molecular subtypes using pulsed-field gel electrophoresis (PFGE). Approximately 42% (61/146) of the isolates were susceptible to all 13 antimicrobials tested. More than half of the isolates (50.70%) were resistant to ampicillin, 49.32% to sulfisoxazole, 17.12% to tetracycline, and 15.75% to doxycycline. Thirty (20.55%) isolates were resistant to three or more antimicrobials. The avrA, mgtC, and sopE virulence genes were identified in all isolates, while 97.2% and 92.4% were positive for bcfC and spvC genes, respectively. Genes associated with resistance to streptomycin (aadA), β-lactams (blaTEM, blaCMY, blaSHV, and blaCTX), tetracycline (tetA and tetB), and sulfonamides (sulI, sulII, and sulIII) were detected among corresponding resistant isolates. A total of 41 PFGE patterns were identified from 77 antimicrobial resistance (AMR) isolates and were primarily grouped into seven clusters (A-G), each with 90% similarity. The majority of Salmonella Enteritidis isolates (63.63%, 49/77) shared the same PFGE cluster, indicating potential cross contamination during processing and cutting or working during retailing and marketing. A significantly (p < 0.05) lower percentage (<25%) of isolates belonging to clusters D and E were resistant to sulfisoxazole compared with those belonging to clusters A, B, C, F, and G (>80%), indicating that sulfisoxazole resistance might be associated with genetic content (PFGE profiles) of Salmonella Enteritidis. This study provides important and updated information about the baseline antimicrobial-resistant data for food safety risk assessment of Salmonella Enteritidis from retailed chicken in Shanghai, which is the first step for the development and implementation of China's AMR National Action Plan, and can be helpful for future surveillance activities to ensure the safety of the chicken supply.

Salmonella Enteritidis remains a standout among the leading causes of foodborne diseases worldwide. Previous studies have demonstrated that a unique clonal group of Salmonella Enteritidis, named SE86, is involved in foodborne outbreaks in southern Brazil and is frequently identified among strains isolated from poultry. The aim of this study was to determine the influence of the isolation source (food products involved in salmonellosis outbreaks and poultry sources) on the phenotypic and molecular characteristics of Salmonella Enteritidis SE86. A biofilm formation assay, antimicrobial susceptibility test, polymerase chain reaction identification of virulence-associated genes, and phage type 4 (PT4) assessment were performed to characterize Salmonella Enteritidis SE86. The human strains presented less antimicrobial resistance than the poultry strains. Resistance to some substances was related to the isolation source of the strain. Strains of the same clonal group presented different biofilm production abilities. Biofilm formation was independent of the isolation source at all temperatures. Temperature influenced biofilm formation only by the poultry strains. Most of the investigated genes presented a high frequency and a regular distribution, regardless of the isolation source. The spvB, spiA, pagC, sipB, prgH, spaN, sitC, and lpfC genes were associated with the avian strains, whereas iroN was associated with the strains isolated from food products involved in salmonellosis outbreaks. Most strains belonged to PT4. No relationship was found between biofilm production and antimicrobial resistance or between the virulence profile and biofilm production or antimicrobial resistance.

In this study, novel antimicrobial peptides had been derived by enzymatic hydrolysis of filefish (Thamnaconus modestus) byproduct (HFBP). Different proteases, (papain [P], trypsin [T], neutrase [N], pepsin [PE], and the mixture I [PN] and mixture II [PT]) and different hydrolysis time (60, 120, 180, and 240 min), have been used to generate peptides with different lengths and amino acid sequences. The antimicrobial properties of HFBPs were tested, against Gram-positive and Gram-negative strains, using disc diffusion method. HFBP prepared after 120 min of the enzymatic hydrolysis by trypsin (HFBP-T) exhibited greatest antibacterial activities. Bacillus cereus 10451 (BC) and Salmonella enteritidis 10982 (SE) strains were most sensitive to HFBP-T with an inhibitory zone of 24.68 and 29.67 mm diameter and minimum inhibitory concentration of 1.25 and 2.5 mg/mL, respectively. Moreover, the antimicrobial activities of tested HFBPs increased significantly at low pH and temperature. The amino acid analysis showed that HFBP-T protein hydrolysate is high in an amino acid of proline, which probably contributes to the antimicrobial activity. The results obtained from scanning electron microscopy suggested that HFBPs might kill bacteria by acting on the cell wall of bacteria. Conclusively, the HFBP derived from filefish byproduct with biological activates is an interesting alternative to the use of waste from the fishing industry as natural antimicrobials in food stuff.

Nontyphoidal Salmonella are one of the leading causes of foodborne diseases in the world. As poultry products are recognized as main sources of human salmonellosis, nontyphoidal Salmonella control has become a global issue for the poultry industry. The increasing antimicrobial resistance in poultry-related nontyphoidal Salmonella serovars is a global matter of concern. By monitoring the evolution of antimicrobial resistance, alternative treatments can be identified and possible restrictions in the treatment of systemic human salmonellosis foreseen. A meta-analysis was conducted to assess the profile and temporal evolution of the antimicrobial resistance of nontyphoidal Salmonella of poultry and human origin in Brazil, isolated in the period from 1995 to 2014. Four databases were researched; twenty-nine articles met the eligibility criteria and were included in the meta-analysis. In the nontyphoidal isolates of poultry origin, the highest levels of antimicrobial resistance were verified for sulfonamides (44.3%), nalidixic acid (42.5%), and tetracycline (35.5%). In the human-origin isolates, the resistance occurred mainly for sulfonamides (46.4%), tetracycline (36.9%), and ampicillin (23.6%). Twenty-two articles described results of antimicrobial resistance specifically for Salmonella Enteritidis, also enabling the individual meta-analysis of this serovar. For most antimicrobials, the resistance levels of Salmonella Enteritidis were lower than those found when considering all the nontyphoidal serovars. In the poultry-origin isolates, a quadratic temporal distribution was observed, with reduced resistance to streptomycin in Salmonella Enteritidis and in all nontyphoidal serovars, and a linear increase of resistance to nalidixic acid in Salmonella Enteritidis. In the human-origin isolates, a linear increase was identified in the resistance to nalidixic acid in Salmonella Enteritidis and in all the nontyphoidal isolates, and to gentamicin in Salmonella Enteritidis. Continuous monitoring of the development and spread of antimicrobial resistance could support the measurement of the consequences on poultry and human health.

The objective of this research was to determine whether variation in the presence of fimbrial protein SefD would impact efficacy of bacterins as measured by recovery of Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) from the spleens of hens. Two bacterins were prepared that varied in SefD content. Also, two adjuvants were tested, namely, water-in-oil and aluminum hydroxide gel (alum). Control groups for both adjuvant preparations included infected nonvaccinated hens and uninfected nonvaccinated hens. At 21 days postinfection, Salmonella Enteritidis was recovered from 69.7%, 53.1%, and 86.0% from the spleens of all hens vaccinated with bacterins lacking SefD, bacterins that included SefD, and infected nonvaccinated control hens, respectively. No Salmonella was recovered from uninfected nonvaccinates. Results from individual trials showed that both bacterins reduced positive spleens, but that the one with SefD was more efficacious. Alum adjuvant had fewer side effects on hens and egg production as compared to water-in-oil. However, adjuvant did not change the relative recovery of Salmonella Enteritidis from spleens. These results suggest that SefD is a promising target antigen for improving the efficacy of immunotherapy in hens, and is intended to reduce Salmonella Enteritidis in the food supply.

Salmonella Enteritidis (SE) is a major foodborne pathogen responsible for causing gastrointestinal infections in humans, predominantly due to the consumption of contaminated eggs. In layer hens, SE colonizes the intestine and migrates to various organs, including the oviduct, thereby leading to egg yolk and shell contamination. This study investigated the efficacy of caprylic acid (CA), a medium-chain fatty acid, in reducing SE colonization and egg contamination in layers. Caprylic acid was supplemented in the feed at 0%, 0.7%, or 1% (vol/wt) from day 1 of the experiment. Birds were challenged with 10(10) log colony-forming units (CFU)/mL of SE by crop gavage on day 10, and re-inoculated (10(10) log CFU/mL) on day 35. After 7 days post first inoculation, eggs were collected daily and tested for SE on the shell and in the yolk separately. The birds were sacrificed on day 66 to determine SE colonization in the ceca, liver, and oviduct. The consumer acceptability of eggs was also determined by triangle test. The experiment was replicated twice. In-feed supplementation of CA (0.7% and 1%) to birds consistently decreased SE on eggshell and in the yolk (p<0.05). Supplementation of CA at 1.0% decreased SE population to ≈14% on the shell and ≈10% in yolk, when compared to control birds, which yielded ≈60% positive samples on shell and ≈43% in yolk. Additionally, SE populations in the cecum and liver were reduced in treated birds compared to control (p<0.05). No significant difference in egg production, body weight, or sensory properties of eggs was observed (p>0.05). The results suggest that CA could potentially be used as a feed additive to reduce eggborne transmission of SE.

The effect of dietary caprylic acid (CA) on Salmonella Enteritidis, as well as the surface treatment of chicken skin contaminated with Salmonella Enteritidis was evaluated. To evaluate the dietary effect of CA on Salmonella Enteritidis, the individually housed broiler chickens (n=48) were divided into 4 groups (positive control, negative control, 2.5 g/kg of CA in the feed, and 5 g/kg of CA in the feed). The feed of all groups, except the negative control, was artificially contaminated with Salmonella Enteritidis ATCC 13076 (10(7) colony-forming units/100 g of feed). Both concentrations of dietary CA significantly decreased counts of Salmonella Enteritidis in the crop and cecum of experimental chickens (p<0.05). The effect of CA in the crop contents was more pronounced than in the cecum. Surface treatment of chilled chicken halves with CA at 1.25 and 2.5 mg/mL significantly decreased Salmonella Enteritidis contamination of chicken skin (p<0.05). The sensory evaluation of the skin and meat showed that treatment of the skin with 1.25 mg/mL of CA worsened odor and appearance of the chicken skin, while sensory traits of chicken meat were not significantly affected. Taste and overall acceptability was not influenced by CA in both meat and skin. Treatment of the skin with 2.5 mg/mL of CA resulted in more pronounced changes of the skin odor and appearance. In conclusion, dietary CA reduced carriage of Salmonella Enteritidis in chickens, whereas surface-treatment reduced or eliminated Salmonella Enteritidis contamination in the processed bird.

Information about nontyphoidal Salmonella (NTS) infection in children is limited in mainland China. The objective of this study was to investigate the prevalence, serotypes, and antibiotic resistance patterns of NTS infection in children in Shanghai. All cases with probable bacterial diarrhea were enrolled from the enteric clinic of a tertiary pediatric hospital between July 2010 and December 2011. Salmonella isolation, serotyping, and antimicrobial susceptibility testing were conducted by the microbiological laboratory. NTS were recovered from 316 (17.2%) of 1833 cases with isolation rate exceeding Campylobacter (7.1%) and Shigella (5.7%). NTS infection was prevalent year-round with a seasonal peak during summer and autumn. The median age of children with NTS gastroenteritis was 18 months. Fever and blood-in-stool were reported in 52.5% and 42.7% of cases, respectively. Salmonella Enteritidis (38.9%) and Salmonella Typhimurium (29.7%) were the most common serovars. Antimicrobial susceptibility showed 60.5% of isolates resistant to ≥1 clinically important antibiotics. Resistance to ciprofloxacin and the third-generation cephalosporins was detected in 5.5% and 7.1%-11.7% of isolates, respectively. NTS is a major enteropathogen responsible for bacterial gastroenteritis in children in Shanghai. Resistance to the current first-line antibiotics is of concern. Ongoing surveillance for NTS infection and antibiotic resistance is needed to control this pathogen in Shanghai.

With the increasing incidence of Salmonella enterica serovar Enteritidis (SE) infections, five SE foodborne outbreaks were identified between 2008 and 2010 on Jeju Island, Republic of Korea. In this study, the genetic relatedness of isolates recovered from the five outbreaks was investigated to identify the source of foodborne SE infections. In total, 57 SE isolates from five outbreaks (17 isolates, 5 isolates, 18 isolates, 8 isolates, and 9 isolates, respectively) were used for antimicrobial susceptibility testing, pulsed-field gel electrophoresis (PFGE), and multiple-locus variable-number tandem repeat analysis (MLVA). SE isolates from 2008 and 2009 were resistant to nalidixic acid, whereas SE isolates from 2010 were resistant to five antibiotics. Of the five outbreaks, outbreaks A, B, and D had identical PFGE-XbaI and PFGE-BlnI patterns, SEGX01.003 and SEGA26.001, respectively. Outbreak C had patterns SEGX01.011 and SEGA26.005, and outbreak E had patterns SEGX01.007 and SEGA26.007. However, MLVA profiles further distinguished the SE isolates from each outbreak into patterns SEGM.014 (outbreak A), SEGM.012 (outbreak B), SEGM.008 (outbreak C), SEGM.016 (outbreak D), and SEGM.015 (outbreak E). Among these five outbreaks, three outbreaks were presumed to be caused by the clonal SE isolates depending on PFGE pattern, but the MLVA results elucidated that these were caused by different SE isolates from the different origins. Therefore, for the epidemiological investigation or surveillance of SE foodborne diseases, both PFGE and MLVA should be used together.

Salmonella enterica serovar Enteritidis is a leading cause of salmonellosis throughout the world and is most commonly associated with the consumption of contaminated poultry and egg products. Salmonella Enteritidis has enhanced ability to colonize and persist in extraintestinal sites within chickens. In this study, 54 Salmonella Enteritidis isolates from human patients (n=28), retail chicken (n=9), broiler farms (n=9), and egg production facilities (n=8) were characterized by antimicrobial susceptibility testing, plasmid analysis, genetic relatedness using XbaI and AvrII pulsed-field gel electrophoresis (PFGE), and the presence of putative virulence genes. Nine isolates were evaluated for their abilities to invade and survive in intestinal epithelial and macrophage cell lines. Overall, 56% (n=30) of isolates were resistant to at least one antimicrobial agent tested, yet no isolates showed resistance to more than three antimicrobials. All isolates carried a common ∼55-kb plasmid, with some strains containing additional plasmids ranging from 3 to 50 kb. PFGE analysis revealed five XbaI and AvrII clusters. There were significant overlaps in the PFGE patterns of the isolates from human, chicken, and egg houses. All isolates tested PCR positive for iacP, purR, ttrB, spi4H, rmbA, sopE, invA, sopB, spvB, pagC, msgA, spaN, orgA, tolC, and sifA, and negative for iss, virB4, and sipB. Of the isolates selected for virulence testing, those containing the iron acquisition genes, iutA, sitA, and iucA, and ∼50-kb plasmids demonstrated among the highest levels of macrophage and epithelial cell invasion, which may indicate their importance in pathogenesis.

Four hundred and thirty Salmonella isolates, recovered from various food-animal products, were tested for nalidixic acid resistance, plasmid-mediated quinolone resistance, and genetic relationship. One hundred fifteen isolates (113 Salmonella serovar Enteritidis and two Salmonella serovar Typhimurium isolates) of 430 (26.7%) Salmonella isolates exhibited nalidixic acid resistance. Polymerase chain reaction screening for qnrA, qnrB, qnrS, qepA (encoding fluoroquinolones resistance) and rmtB (encoding aminoglycosides resistance) showed that 5 (1.16%) isolates were positive for qnr- and qepA-type genes, and the aac(6')-Ib-cr gene was observed in two (1.7%) Enteritidis isolates concomitantly with qnrA or qnrB. The co-occurrence of qepA and rmtB in one Typhimurium isolate is noteworthy. Pulsed-field gel electrophoresis revealed a high genetic homogeneity of nalidixic-resistant isolates and the persistence of clonal clusters over 4 years in different regions in Tunisia and from various food-animal products. To the best of our knowledge, this is the first report of co-occurrence of qepA and rmtB in a Salmonella strain.

Most bacterial pathogens associated with human enteric illness have zoonotic origins and can be transmitted directly from animals to people or indirectly through food and water. This multitude of potential exposure routes and sources makes the epidemiology of these infectious agents complex. To better understand these illnesses and identify solutions to reduce human disease, an integrative approach like One Health is needed. This article considers the issue of Salmonella in Canada and interprets data collected by several Canadian surveillance and research programs. We describe recovery of Salmonella from various samples collected along the exposure pathway and compare the serovars detected in the different components under surveillance (animal, food, environment, and human). We then present three examples to illustrate how an approach that interprets multiple sources of surveillance data together is able to address issues that transcend multiple departments and jurisdictions. First, differences observed in recovery of Salmonella from different cuts of fresh chicken collected by different programs emphasize the importance of considering the surveillance objectives and how they may influence the information that is generated. Second, the high number of Salmonella Enteritidis cases in Canada is used to illustrate the importance of ongoing, concurrent surveillance of human cases and exposure sources to information domestic control and prevention strategies. Finally, changing patterns in the occurrence of ceftiofur-resistant Salmonella Heidelberg in retail meats and humans demonstrates how integrated surveillance can identify an issue in an exposure source and link it to a trend in human disease. Taken together, surveillance models that encompass different scales can leverage infrastructure, costs, and benefits and generate a multidimensional picture that can better inform disease prevention and control programs.

Non-typhoidal salmonellosis is an important zoonotic disease caused by Salmonella enterica. This work focuses on the identification of Salmonella enterica clonal strains which, presenting a wide distribution potential, express resistance determinants that compromise effectiveness of the antimicrobial therapy. The screening was performed on 506 Salmonella enterica isolates from animals and humans, which were characterized by serovar and phage typing, genome macrorestriction and pulsed-field gel electrophoresis, and detection of phenotypic and genotypic traits for antimicrobial resistance. A Salmonella Enteritidis strain with strong quinolone resistance is spread on three host environments carrying one of the four variants found for the GyrA protein: (1) Asp87Tyr, the major polymorphism found in 39 Salmonella isolates from human origin and six from poultry; (2) Ser83Phe, with four isolates from human origin and one from white stork (Ciconia ciconia); and (3) Asp87Asn or (4) Asp87Gly, with two isolates each from human origins. Several Salmonella Typhimurium strains that presented int1 elements and the classically associated pentaresistance (ACSSuT) phenotype were found distributed between two host environments: domestic animals and humans, domestics and wild animals, or wild fauna plus humans. This study points out the importance of monitoring gut microbiota and its antimicrobial resistance from wildlife, in parallel to livestock animals and humans, especially for animal species that are in close contact with people.

The objective of this research was to determine minimal inhibitory concentration (MIC) population distributions for colistin for Salmonella on subtype level. Furthermore, we wanted to determine if differences in MIC for colistin could be explained by mutations in pmrA or pmrB encoding proteins involved in processes that influence the binding of colistin to the cell membrane. During 2008-2011, 6,583 Salmonella enterica subsp. enterica isolates of human origin and 1931 isolates of animal/meat origin were collected. The isolates were serotyped, and susceptibility was tested towards colistin (range 1-16 mg/L). Moreover, 37 isolates were tested for mutations in pmrA and pmrB by polymerase chain reaction (PCR) and DNA sequencing. MIC distribution for colistin at serotype level showed that Salmonella Dublin (n=198) followed by Salmonella Enteritidis (n=1247) were less susceptible than "other" Salmonella serotypes originating from humans (n=5,274) and Salmonella Typhimurium of animal/meat origin (n=1794). MIC was ≤1 mg/L for 98.9% of "other" Salmonella serotypes originating from humans, 99.4% of Salmonella Typhimurium, 61.3% of Salmonella Enteritidis, and 12.1% of Salmonella Dublin isolates. Interestingly, Salmonella Dublin and Salmonella Enteritidis belong to the same O-group (O:1, 9,12), suggesting that surface lipopolysaccharides (LPS) of the cell (O-antigen) play a role in colistin susceptibility. The epidemiological cut-off value of >2 mg/L for colistin suggested by European Committee on Antimicrobial Susceptibility Testing (EUCAST) is placed inside the distribution for both Salmonella Dublin and Salmonella Enteritidis. All tested Salmonella Dublin isolates, regardless of MIC colistin value, had identical pmrA and pmrB sequences. Missense mutations were found only in pmrA in one Salmonella Reading and in pmrB in one Salmonella Concord isolate, both with MIC of ≤1 for colistin. In conclusion, our study indicates that missense mutations are not necessarily involved in increased MICs for colistin. Increased MICs for colistin seemed to be linked to specific serotypes (Salmonella Dublin and Salmonella Enteritidis). We recommend that Salmonella with MIC of >2 mg/L for colistin be evaluated on the serovar level.

Salmonella is one of the most common foodborne pathogens in humans. Laboratory-based surveillance for non-typhoidal Salmonella infection was conducted in Guangdong Province, China to improve understanding about the disease burden and detection of dispersed outbreaks. Salmonella isolated from patients with diarrhea were sent from 16 sentinel hospitals to local public health laboratories for confirmation, serotyping, antimicrobial susceptibility testing, and pulsed-field gel electrophoresis (PFGE). PFGE patterns were analyzed to identify clusters representing potential outbreaks. Between September 2009 and October 2010, 352 (4%) Salmonella isolates were obtained from 9167 stool specimens. Salmonella enterica serotype Typhimurium (45%) and Salmonella enterica serotype Enteritidis (13%) were the most common serotypes, and multidrug resistance was high, especially in Salmonella Typhimurium isolates. PFGE patterns of obtained Salmonella isolates were found to be diverse, but a unique PFGE pattern comprising 53 Salmonella Typhimurium isolates were found to occur almost exclusively in infants. Epidemiologic studies are ongoing to determine whether a common exposure is the source of the Salmonella Typhimurium strain frequently isolated from infants.

In the United States, serovar Kentucky has become one of the most frequently isolated Salmonella enterica serovars from chickens. The reasons for this prevalence are not well understood. Phenotypic comparisons of poultry Salmonella isolates belonging to various serovars demonstrated that serovar Kentucky isolates differed from those of most other serovars in their response to acid. Microarray and qPCR analyses were performed with aerated exponentially growing poultry isolates, Salmonella enterica serovar Kentucky 3795 and Enteritidis Nal(R), exposed for 10 min to tryptic soy broth (TSB) adjusted to pH 4.5 with HCl and to pH 5.5 with HCl or acetic acid. Data obtained by microarray analysis indicated that more genes were up- or down-regulated in strain Kentucky 3795 than in Enteritidis Nal(R) under acidic conditions. Acid exposure in general caused up-regulation of energy metabolism genes and down-regulation of protein synthesis genes, particularly of ribosomal protein genes. Both strains appear to similarly utilize the lysine-based pH homeostasis system, as up-regulation of cadB was observed under the acidic conditions. Expression of regulatory genes (rpoS, fur, phoPQ) known to be involved in the acid response showed similar trends in both isolates. Differences between Kentucky 3795 and Enteritidis Nal(R) were observed with respect to the expression of the hdeB-like locus SEN1493 (potentially encoding a chaperone important to acid response), and some differences in the expression of other genes such as those involved in citrate utilization and motility were noted. It appears that the early stages of the transcriptional response to acid by isolates Kentucky 3795 and Enteritidis Nal(R) are similar, but differences exist in the scope and in some facets of the response. Possibly, the quantitative differences observed might lead to differences in protein levels that could explain the observed differences in the acid phenotype of serovar Kentucky and other Salmonella serovars.

Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major serovar associated with human salmonellosis. A total of 425 clinical S. Enteritidis isolates of human origin were collected between June 2009 and September 2010 from North Carolina. The isolates were further characterized for antimicrobial susceptibility, antimicrobial resistance coding determinants, virulence genes, and fingerprint profiles to determine whether they were similar or different to the S. Enteritidis strain responsible for the human outbreak due to consumption of contaminated eggs. Ten different antimicrobial resistance phenotypes were observed with the highest frequency of resistance exhibited to ampicillin (n=10; 2.35%). The isolates were predominantly pansusceptible (n=409; 96.23%); however, seven isolates were multidrug resistant (MDR; i.e., resistant to three or more antimicrobials). Extended spectrum β-lactamase (ESBL) coding genes (bla(TEM) and bla(PSE)) were detected in the ampicillin-resistant isolates, whereas a single MDR isolate tested positive for class 1 integron (1 kb). The majority of the isolates (n=422; 99.3%) carried the invA, mgtC, stn, sopB, sopE1, and sefA virulence genes. However, 37 (8.7%) and 46 (10.82%) S. Enteritidis isolates tested negative for the plasmid encoded genes spvC and rck, respectively. Pulsed-field gel electrophoresis (PFGE) typing of 118 S. Enteritidis isolates by restriction enzymes XbaI and BlnI resulted in seven clusters, each with a discriminatory index (DI) of 0.715 and 0.785, respectively. The combination of XbaI-BlnI patterns generated a dendrogram with 14 clusters and a higher DI of 0.914. The PFGE profile of 80 isolates matched 100% with the S. Enteritidis strain that has been cited for the recent outbreak in the United States due to consumption of contaminated eggs. In conclusion, we identified a genotypic similar S. Enteritidis population in our study based on antimicrobial susceptibility, virulence gene, and PFGE fingerprint profiles.

Background: Salmonella enterica serotype Enteritidis is a universally recognized cause of foodborne disease. In South Africa, outbreaks of foodborne disease are generally under reported. We investigated the etiology of acute gastroenteritis in 216 patients who presented to a rural hospital in KwaZulu-Natal, South Africa, after consuming a meal at a school function., Materials and Methods: Stool specimens from 37 patients, as well as two food samples, were available for microbiological investigation. Similarity between isolates was investigated using phenotypic and genotypic techniques. Phenotypic investigations included morphological, biochemical, and antibiogram profiling. Genotypic relatedness was determined with pulsed-field gel electrophoresis analysis. The available epidemiological data were also described., Results: Salmonella Enteritidis was isolated from 18 patients and 1 food sample. Isolates were phenotypically and genotypically indistinguishable. Epidemiological data suggest a point-source outbreak with a possibility of continued transmission., Conclusions: The results suggest a foodborne Salmonella Enteritidis outbreak due to contaminated food served at the school function. Epidemiological investigations continue to be extremely difficult in rural areas.

The present study was conducted to investigate serotype distribution, antimicrobial resistance patterns, carriage of class 1 integron, and clonality of Salmonella strains isolated from patients aged 0-12 years in Tehran, Iran, during 2007-2008. A total of 139 Salmonella isolates were studied. Salmonella serotypes Enteritidis, Infantis, and Typhimurium included 84.9% of isolates, Enteritidis accounting for 41.7%. The most prevalent resistances were to doxycycline (64.7%), nalidixic acid (61.2%), tetracycline (51.8%), and streptomycin (42.8%). Fifty-three (38.1%) isolates contained class 1 integron. Eight different gene cassettes were identified, aadA1 being the most frequently encountered. Pulsed-field gel electrophoresis showed that integron-positive Salmonella strains belonging to serotypes Infantis, Enteritidis, and Typhimurium were attributed to two, three, and five different pulsotypes, respectively. The findings indicated that the distribution and drug resistance pattern of most prevalent Salmonella serotypes were broadly similar to that reported globally from human isolates. Presence of class 1 integrons was common among Salmonella serotypes in Tehran, Iran. Concurrent clonal expansion and horizontal transmission events seem to contribute to increase in drug resistance prevalence among Salmonella serotypes., (© Mary Ann Liebert, Inc.)

In this article, the sensitivity of Salmonella enteritidis to surfactin and polylysine was observed, and the optimization of antimicrobial activity of surfactin and polylysine against S. enteritidis in milk was evaluated by a response surface methodology. Results showed that S. enteritidis was very sensitive to them, whose minimal inhibitory concentrations were 6.25 and 31.25 μg/mL, respectively. The optimization result indicated that S. enteritidis could be sterilized by 6 orders of magnitude when the temperature was 4.45°C, the action time was 6.91 h, and the concentration (surfactin/polylysine weight ratio, 1:1) was 10.03 μg/mL.

Molecular epidemiology at local scale in Sicily (Italy) of ampicillin resistance in Salmonella spp. isolates from municipal wastewater (n = 64) and clinical specimens (n = 274) is described in comparison with previously examined Escherichia coli isolates (n = 273) from wastewater. High prevalence of antibiotic resistance (28.9%) with highest resistance rates against ampicillin (22.7%) was observed in E. coli isolates. Different resistance rates were observed in Salmonella according to the serovars, with prevalences of the same order in both wastewater and clinical isolates belonging to the same serovar (e.g., 91.7% ampicillin resistance in wastewater isolates vs. 70.8% in clinical isolates of the Salmonella serovar Typhimurium and 0% ampicillin resistance in both wastewater and clinical isolates of the Salmonella serovar Enteritidis). The beta-lactam resistance gene bla(TEM) was present in both wastewater and clinical Salmonella spp. isolates, with the exception of Salmonella enterica serovar Typhimurium isolates with a typical six-drug resistance pattern AmpChlSulTeStrSp that had the bla(PSE-1) gene. The bla(TEM) gene was present in all the E. coli isolates but one had the bla(SHV) gene. Several E. coli and some Salmonella isolates were positive for class 1 integrons with variable regions of 1.0 or 1.5 kb containing aadA1, dfrA17-aadA5, or dfrA1-aadA1 gene cassettes, whereas Salmonella serovar Typhimurium isolates with the six-drug resistance pattern were positive for both 1.0 and 1.2 kb integrons. Polymerase chain reaction replicon typing demonstrated the presence of multireplicon resistance plasmids in several isolates of E. coli, containing two to four of the replicons IncF, IncI1, IncFIA, and IncFIB, whereas other isolates showed resistance plasmids with only IncF, IncP, or IncK replicons. Replicon IncI1 was detected in one Salmonella isolate, whereas other isolates belonging to different serovars had IncN replicons. Analysis of isolates from wastewater can be a useful epidemiologic tool to monitor the prevalence of antibiotic resistance and genetic elements related to antibiotic resistance in Salmonella clones circulating in the human population.

Aim: To examine the prevalence of antimicrobial-resistant Salmonella in chicken meat and correlate with isolates from ill humans., Methods: We isolated Salmonella from raw chicken purchased from a randomly selected sample of retail outlets in central Pennsylvania during 2006-2007. Salmonella isolates from meat were compared, using pulsed-field gel electrophoresis, to isolates in the PulseNet database of Salmonella recovered from humans., Results: Of 378 chicken meat samples, 84 (22%) contained Salmonella. Twenty-six (31%) of the Salmonella isolates were resistant to > or = 3 antimicrobials and 18 (21%) were resistant to ceftiofur. All ceftiofur-resistant isolates exhibited reduced susceptibility (minimum inhibitory concentration >2 microg/mL) to ceftriaxone and carried a bla(CMY) gene, as detected by polymerase chain reaction. Among the 28 Salmonella serovar Typhimurium isolates, 20 (71.4%) were resistant to > or = 3 antimicrobials and 12 (42.9%) were resistant to ceftiofur. One ceftiofur-resistant Salmonella serovar Typhimurium poultry isolate exhibited a rare pulsed-field gel electrophoresis pattern indistinguishable from a human isolate in PulseNet; both isolates carried the bla(CMY-2) gene., Conclusions: These data demonstrate the presence of multidrug-resistant Salmonella in poultry meat, including bla(CMY) plasmid-mediated genes that confer resistance to both ceftiofur, used in poultry, and ceftriaxone, used for treating salmonellosis in humans. This study illustrates the potential for molecular subtyping databases to identify related Salmonella isolates from meat and ill humans, and suggests that chicken could be a source for multidrug-resistant salmonellosis in humans.

A majority of the seed sprout-related outbreaks have been associated with Escherichia coli O157:H7 and Salmonella spp. Therefore, it is necessary to find an effective method to inactivate these microorganisms on the seeds before sprouting. When treatment with hot water at 85 degrees C for 40 sec followed by dipping in cold water for 30 sec and soaking into chlorine water (2000 ppm) for 2 h was performed, no viable pathogens were found in the enrichment medium and during the sprouting process. The germination yield of the seed was not affected significantly (p > 0.05). Therefore, these treatments could be useful for the decontamination method of mung bean seeds intended for sprout production.

The BT/TAMUS 2032 (BT) cationic peptides are a group of related cationic peptides produced by a Gram-positive soil bacterium, Brevibacillus texasporus. Cationic amphiphilic peptides produced by host cells have been found to stimulate or prime the innate immune responses in mammals, but little information is available on the effects of bacterial-produced peptides on host immunity. We have previously shown that BT, provided as a feed additive for 4 days after hatch, significantly induced protection against extraintestinal colonization by Salmonella enterica serovar Enteritidis. We also found that feeding BT significantly upregulated the functional efficiency of heterophils, the avian equivalent to mammalian neutrophils. The objective of the present study was to further evaluate the effect of BT as a nonantibiotic, antibacterial compound and a stimulator of the innate immune response of young chickens. BT, provided as a feed additive at three different concentrations (12, 24, or 48 ppm) for 4 days after hatch, significantly increased protection against Salmonella enterica serovar Enteritidis cecal colonization in a concentration-dependent manner. We also confirmed our previous results that the functional activities of heterophils from chickens fed the BT rations were significantly upregulated. In addition, we also found that the functional activities of peripheral blood monocytes were significantly increased in a concentration-dependent manner when compared with monocytes isolated from chickens fed a control diet. This is the first report of bacterial cationic peptides providing protection against Salmonella cecal colonization. The significance of these data is that the orally delivered cationic peptides stimulate the innate response during the first week after hatch, normally a time of immunologic inefficiency and increased susceptibility to bacterial infections. We speculate that BT given as a feed additive during the first week after hatch could provide increased protection against a variety of bacterial pathogens because of the nonspecific nature of the innate response.

The antimicrobial activity of essential oils of oregano, thyme, basil, marjoram, lemongrass, ginger, and clove was investigated in vitro by agar dilution method and minimal inhibitory concentration (MIC) determination against Gram-positive (Staphylococcus aureus and Listeria monocytogenes) and Gram-negative strains (Escherichia coli and Salmonella Enteritidis). MIC(90%) values were tested against bacterial strains inoculated experimentally in irradiated minced meat and against natural microbiota (aerobic or facultative, mesophilic, and psychrotrophic bacteria) found in minced meat samples. MIC(90%) values ranged from 0.05%v/v (lemongrass oil) to 0.46%v/v (marjoram oil) to Gram-positive bacteria and from 0.10%v/v (clove oil) to 0.56%v/v (ginger oil) to Gram-negative strains. However, the MIC(90%) assessed on minced meat inoculated experimentally with foodborne pathogen strains and against natural microbiota of meat did not show the same effectiveness, and 1.3 and 1.0 were the highest log CFU/g reduction values obtained against tested microorganisms.

The protective effects of powder preparation of egg yolk immunoglobulin Y (IgY), specific to Salmonella Enteritidis and Salmonella Typhimurium outer membrane proteins (OMP), against these two Salmonella sp. serovars were investigated in vitro in two different assays: adhesion-prevention and growth-inhibition. The adhesion-prevention assay was conducted using polarized monolayers of the human intestinal epithelial Caco-2 cell line. First, the conditions of Salmonella adherence to Caco-2 cells were optimized, and interferences of bacteria with the transepithelial electrical resistance (TER) of fully differentiated Caco-2 cell monolayers and the lactate dehydrogenase release upon exposure of the cells to Salmonella were evaluated. Both Salmonella sp. serovars were able to adhere to Caco-2 cells and decreased TER. Results from the adhesion-prevention assay demonstrated that specific IgY reduced the decrease in TER of the infected Caco-2 cell monolayers and blocked the Salmonella sp. adhesion in a concentration-dependent manner (p < 0.05). Nonspecific IgY also exhibited an inhibitory effect on these two parameters, but to a lesser extent than that of the specific IgY (p < 0.05). The protective effect of nonspecific IgY could be attributed to the low-density lipoprotein component of the water-soluble fraction of egg yolks that may not have been eliminated during ultrafiltration. The growth-inhibition assay revealed that specific IgY had an inhibitory effect on the bacterial growth, markedly during the late exponential phase, whereas nonspecific IgY failed to do so. Taken together, these results suggest that the in vitro growth inhibitory effect of specific IgY on Salmonella spp. resulted from the specific binding activity of these IgY to Salmonella sp. OMP. Passive immunization with Salmonella sp. OMP-specific IgY could thus be useful to prevent Salmonella colonization in broiler chickens and the subsequent carcass contamination during processing.

Thirty-eight Salmonella Enteritidis isolates from chickens and chicken meat in Turkey were examined for antimicrobial susceptibility, XbaI pulsed-field gel electrophoresis (PFGE) patterns, phage types, plasmid profiles, and resistance genes. Seven different PFGE patterns were observed, with the most common accounting for 71% (X1). The most common phage type was PT4, followed by PT7, PT16, PT1, PT6, and PT35. Different phage types shared the same PFGE pattern. Twenty-one isolates were susceptible to all antimicrobial agents tested whereas eight were resistant to two or more antimicrobials. Six isolates were resistant to gentamicin, spectinomycin, streptomycin, and sulphamethoxazole and one of these in addition to nalidixic acid. Two isolates were resistant to ampicillin and nalidixic acid. An additional nine isolates were resistant to nalidixic acid only. All six streptomycin-resistant isolates had aadA located in an integron class 1 structure. Both ampicillin-resistant isolates had the bla(TEM) gene. Five different plasmid profiles were found among the isolates. Sixty-five percent of isolates contained a single plasmid with an approximate size of 55-60 kb. Plasmid profiling confirmed the PFGE pattern.

The diversity among 1354 strains of Salmonella enterica subsp. enterica, serotype Enteritidis (n = 847) and Typhimurium (n = 507) isolated in Finland in 1991-2002 (n = 608) and in 2003 (n = 746) were studied. The former strains were studied retrospectively by phage typing and pulsed-field gel electrophoresis (PFGE) harmonized in the European Salm-gene project. The latter strains were studied prospectively, and the results correlated to their antimicrobial susceptibility and association with travel to popular tourist destinations. During both periods, S. Enteritidis phage types (PTs) PT1 and PT4, and S. Typhimurium definite types (DTs) DT1 and DT104 were the major phenotypes. SENTXB.0001 was the dominating single PFGE type among S. Enteritidis strains (40% in 1991-2002; 57% in 2003), and accounted correspondingly for 23% and 63% of the PT1 strains, and 81% and 88% of the PT4 strains. No PFGE types dominated among the S. Typhimurium strains but a correlation was found between certain phage and PFGE types: among DT1 strains, STYMXB.0098 accounted for 66% (1991-2002) and 98% (2003) and among the DT104 strains STYMXB.0001 accounted for 84% and 97% in the two time periods, respectively. Of the S. Enteritidis strains isolated in 2003, 91% were associated with travel, most commonly to Spain, Greece, and Bulgaria. SENTXB.0001 was the major Salmonella PFGE type in these countries. In contrast, most (55%) S. Typhimurium strains were of domestic origin. While only 1.3% of the S. Enteritidis strains were multiresistant and 24% were resistant to nalidixic acid only, 30% of the S. Typhimurium strains were multiresistant. Among the multiresistant S. Typhimurium strains, R-type ACSSuT and PFGE type STYMXB.0001 of the DT104 complex dominated.