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A One Health Perspective on Salmonella enterica Serovar Infantis, an Emerging Human Multidrug-Resistant Pathogen
Abstract
Salmonella enterica serovar Infantis presents an ever-increasing threat to public health because of its spread throughout many countries and association with high levels of antimicrobial resistance (AMR). We analyzed whole-genome sequences of 5,284 Salmonella Infantis strains from 74 countries, isolated during 1989-2020 from a wide variety of human, animal, and food sources, to compare genetic phylogeny, AMR determinants, and plasmid presence. The global Salmonella Infantis population structure diverged into 3 clusters: a North American cluster, a European cluster, and a global cluster. The levels of AMR varied by Salmonella Infantis cluster and by isolation source; 73% of poultry isolates were multidrug resistant, compared with 35% of human isolates. This finding correlated with the presence of the pESI megaplasmid; 71% of poultry isolates contained pESI, compared with 32% of human isolates. This study provides key information for public health teams engaged in reducing the spread of this pathogen.
... The results of the serovar prediction reveal a wide variety of serovars circulating in chicken meat sold in wet markets in Metro Manila. It showed the dominance of S. Infantis among the studied Salmonella isolates, which is consistent with previous studies in chicken meat in the U.S., Europe, and Asia (McMillan et al., 2022;Mora et al., 2024;Kim et al., 2024;Mattock et al., 2024). One possible reason for the increased detection of S. Infantis is the presence of plasmid of emerging S. Infantis or pESI, which has been detected in poultry, chicken meat, and clinical isolates across South America, Europe, Africa, and Asia (McMillan et al., 2022;Mattock et al., 2024). ...
... It showed the dominance of S. Infantis among the studied Salmonella isolates, which is consistent with previous studies in chicken meat in the U.S., Europe, and Asia (McMillan et al., 2022;Mora et al., 2024;Kim et al., 2024;Mattock et al., 2024). One possible reason for the increased detection of S. Infantis is the presence of plasmid of emerging S. Infantis or pESI, which has been detected in poultry, chicken meat, and clinical isolates across South America, Europe, Africa, and Asia (McMillan et al., 2022;Mattock et al., 2024). Other significant serovars detected in our study include Brancaster, Anatum, London, and Uganda, all of which have been previously detected in either poultry or poultry products (Xu et al., 2021;Khoo et al., 2023;Mora et al., 2024;Sodagari et al., 2023). ...
... Particularly, most isolates belonged to ST 32 (S. Infantis, eBG 31), consistent with the study of Mattock et al. (2024), where 99% (n = 5,205) of S. Infantis were classified as ST 32, and only few belong to ST 2283 and ST 2146. Other serovars belong to unique STs and eBGs, except for S. Derby, which was linked to STs 40 and 71. ...
The emergence of multidrug-resistant (MDR) Salmonella is recognized as a significant public health problem worldwide. This study investigated the occurrence of MDR Salmonella serovars in chicken meat from wet markets in Metro Manila, Philippines from February to July 2022. Using whole genome sequencing (WGS) and phenotypic antimicrobial resistance (AMR) testing, the serovar, drug resistance, and virulence profiles of Salmonella isolates were characterized. Out of 253 chicken cut samples, 95 S. enterica isolates representing 15 distinct serovars were recovered. The most common was S. enterica serovar Infantis (51.58%), followed by S. Brancaster (9.47%), S. Anatum (7.37%), S. London (7.37%), S. Uganda (6.32%), and S. Derby (4.21%). Phenotypic AMR testing revealed that 73.68% of the isolates were resistant to at least one drug class, and 45.26% were MDR. A wide array of antimicrobial resistance genes (ARGs) associated with resistance to 12 different drug classes was identified, including three β-lactamase gene variants: blaCTX-M-65, blaTEM-1, and blaTEM-176. Some of these ARGs were located on MDR plasmids, such as those on IncFIB(K)_1_Kpn3, IncFIA(HI1)_1_HI1, and IncX1_1. A total of 131 virulence genes were detected, some of which conferred pESI-like characteristics to S. Infantis. These findings highlight a potential public health risk posed by pathogenic MDR Salmonella in chicken meat and underscore the urgent need for further research and coordinated AMR surveillance in the Philippines, aiming to stimulate national efforts to combat AMR.
... The emergence of antimicrobial resistance (AMR) among NTS is concerning as it increases the morbidity, mortality, and cost of treatments (Salam et al., 2023). An increase in antimicrobial-resistant S. Infantis has been detected globally (Mejia et al., 2021;Mattock et al., 2024), including resistance to critically important third-generation cephalosporins and quinolones (Alvarez et al., 2023). Recently, a rise in multidrug resistance (MDR) among S. Infantis isolates has been described, which was linked with the emergence of an IncFIB-like plasmid commonly referred to as the 'plasmid of emerging S. enterica Infantis' (pESI). ...
... The emergence of AMR and MDR among poultry-origin S. Infantis and its transmission through the food chain presents a public health threat (Mattock et al., 2024). To mitigate the health burden of S. Infantis, a One Health approach is needed to consider the complex relationships among human, animal, and environmental health. ...
... In this study, the prevalence of chicken-origin S. Infantis isolates from different sampling sources (retail, cecal, and HACCP) was the highest (16-19 %) compared to all other species, including swine (0-10 %), cattle (1-7 %), turkey (0.3-5 %), and humans (4 %). These findings agree with previous studies from the US (Lee et al., 2022;Mujahid et al., 2023;2023;Sodagari et al., 2024) and globally (Bearson et al., 2024;Iwabuchi et al., 2011;Mattock et al., 2024) where it was shown that S. Infantis became an important serovar among chickens. On the other hand, S. Infantis is not a prevalent serovar in swine and cattle, where the most common serovars are, S. Derby and S. Typhimurium in swine, and S. Enteritidis, S. Schleissheim, and S. Dublin, in cattle, respectively (Montone et al., 2023). ...
... The emergence of antimicrobial resistance (AMR) among NTS is concerning as it increases the morbidity, mortality, and cost of treatments (Salam et al., 2023). An increase in antimicrobialresistant S. Infantis has been detected globally (Mejia et al., 2021;Mattock et al., 2024), including resistance to critically important third-generation cephalosporins and quinolones (Alvarez et al., 2023). Recently, a rise in multidrug resistance (MDR) among S. Infantis isolates has been described, which was linked with the emergence of a large mega plasmid named This preprint research paper has not been peer reviewed. ...
... The emergence of AMR and MDR among poultry-origin S. Infantis and its transmission through the food chain presents a public health threat (Mattock et al., 2024). To mitigate the health burden of S. Infantis, a One Health approach is needed to consider the complex relationships among human, animal, and environmental health. ...
... The MDR among human isolates reported in this study was lower than in the EU (50%) (EFSA, 2023) and globally (25-56%) (Alvarez et al., 2023). However, an increase in MDR isolates is expected as in recent years, S. Infantis has become an emerging serotype, showing high AMR and MDR and a broad host range globally (Mejia et al., 2021;Mattock et al., 2024). Also, in all species, a large proportion of S. Infantis harbors the pESI plasmid, making it MDR (Lee et al., 2021). ...
... Serotype Infantis is also commonly associated with poultry and poultry meat products globally (Alvarez et al. 2023, Mattock et al. 2024. Infantis was the most commonly isolated serotype from broiler chickens in the European Union in 2022, but was among the top four isolated serotypes in the other animal commodities as well ("The European Union One Health 2022 Zoonoses Report" 2023). ...
... This plasmid has also recently expanded into other serotypes, including Senftenberg, Alachua, Muenchen, and others (Cohen et al. 2022, Dos Santos et al. 2022, McMillan et al. 2023, Franciosini et al. 2024. While the plasmid was originally isolated in Israel, it has since been identified all over the world (Mattock et al. 2024). This plasmid contains numerous factors that could contribute to persistence in an avian host, such as fimbriae and siderophores, as well as genes conferring resistance to up to ten antibiotics (Tate et al. 2017, McMillan et al. 2020. ...
Salmonella is a common cause of human foodborne illness, which is frequently associated with consumption of contaminated or undercooked poultry meat. Serotype Infantis is among the most common serotypes isolated from poultry meat products globally. Isolates of serotype Infantis carrying the pESI plasmid, the most dominant strain of Infantis, have been shown to exhibit oxidizer tolerance. Therefore, 16 strains of Salmonella with and without pESI carriage were investigated for susceptibility to biocide chemical processing aids approved for use in US poultry meat processing: peracetic acid (PAA), cetylpyridinium chloride (CPC), calcium hypochlorite, and sodium hypochlorite. Strains were exposed for 15 s to simulate spray application and 90 min to simulate application in an immersion chiller. All strains tested were susceptible to all concentrations of PAA, CPC, and sodium hypochlorite when applied for 90 min. When CPC, calcium hypochlorite, and sodium hypochlorite were applied for 15 s to simulate spray time, strains responded similarly to each other. However, strains responded variably to exposure to PAA. The variation was not statistically significant and appears unrelated to pESI carriage. Results highlight the necessity of testing biocide susceptibility in the presence of organic material and in relevant in situ applications.
... In particular, the ST897 is associated also with the serovars S. kimuenza, an infrequent serotype found in humans, livestock and poultry [44][45][46]. Furthermore, the strains of S. infantis all belonged to ST32, considered the dominant MLST type of this serovar [47]. A recent study based on genomes deposited in a public database revealed that, in Europe, this ST represents more than 97% of strains and 99% worldwide [47]. ...
... Furthermore, the strains of S. infantis all belonged to ST32, considered the dominant MLST type of this serovar [47]. A recent study based on genomes deposited in a public database revealed that, in Europe, this ST represents more than 97% of strains and 99% worldwide [47]. ...
In the European Union, salmonellosis is one of the most important zoonoses reported. Poultry meat and egg products are the most common food matrices associated with Salmonella presence. Moreover, wild and domestic animals could represent an important reservoir that could favour the direct and indirect transmission of pathogens to humans. Salmonella spp. can infect carnivorous or omnivorous wild birds that regularly ingest food and water exposed to faecal contamination. Birds kept in captivity can act as reservoirs of Salmonella spp. following ingestion of infected prey or feed. In this paper, we describe the isolation of different Salmonella serovars in several species of raptors hosted in aviaries in an Italian wildlife centre and in the raw chicken necks used as their feed but intended for human consumption. Characterisations of strains were carried out by integrating classical methods and whole genome sequencing analysis. The strains of S. bredeney isolated in poultry meat and birds belonged to the same cluster, with some of them being multidrug-resistant (MDR) and carrying the Col(pHAD28) plasmid-borne qnrB19 (fluoro)quinolone resistance gene, thus confirming the source of infection. Differently, the S. infantis found in feed and raptors were all MDR, carried a plasmid of emerging S. infantis (pESI)-like plasmid and belonged to different clusters, possibly suggesting a long-lasting infection or the presence of additional undetected sources. Due to the high risk of fuelling a reservoir of human pathogens, the control and treatment of feed for captive species are crucial.
... These isolates are primarily associated with foodborne outbreaks and are linked to contaminated poultry and pork products [90,91]. Moreover, S. Infantis, a serovar increasingly recognized for its MDR, has been reported in Romania, as well as other European countries [92]. ...
This study aims to characterize antibiotic resistance (AR) and virulence markers in Salmonella spp. isolated from Romanian outpatients' stool samples. Methods: In 2019, community-acquired Salmonella strains were collected and identified using MALDI-TOF mass spectrometry, antibiotic susceptibility profiles have been determined with the MicroScan system, and soluble virulence factors were evaluated using specific culture media, while biofilm formation was quantified in 96-well plates. Molecular analysis targeted resistance genes for β-lactams (e.g., bla TEM and bla SHV); tetracyclines (e.g., tet(A)); sulphonamides; and quinolones, as well as virulence genes (e.g., invA, spvC, pldA, and held). Whole-genome sequencing (WGS) was performed on 19 selected isolates. A silver nanoparticles (AgNPsol) alternative to conventional antibiotics was tested for effectiveness against multidrug-resistant (MDR) isolates. Results: From the total of 309 Salmonella isolates (65.05% from children under 4 years of age) belonging to four subtypes and four serovars, 27.86% showed resistance to at least one antibiotic, most frequently to tetracycline, ampicillin, and piperacillin. The strains frequently expressed haemolysin (67%), aesculinase (65%), and gelatinase (62%). Resistance to trimethoprim-sulfamethoxazole was encoded by the sul1 gene in 44.83% of the strains and to tetracyclines by the tet(A) gene (59.52%). The ESBL genes bla TEM , bla SHV , and bla CTX-M were detected by PCR in 16.18%, 2.91%, and 0.65% of the strains, respectively. Additionally, 98.63% of the strains carried the invA marker, with notable positive associations between bla SHV , qnrB, and sul1 with spvC. Conclusions: The present findings revealed significant patterns in Salmonella isolates, subtypes, serovars, AR, Antibiotics 2025, 14, 46 https://doi.org/10.3390/antibiotics14010046 Antibiotics 2025, 14, 46 2 of 34 and virulence, emphasising the need for continuous surveillance of Salmonella infections. Additionally, the potential of AgNPs as an alternative treatment option was demonstrated, particularly for paediatric S. enterica infections.
... Recently, reports of S. Infantis infections in both poultry and human have increased in many countries. A 167 % increase in human infections with S. Infantis was observed in the United States between 2001 and 2016 (Mattock et al., 2024). S. Infantis is the second most common serovar, accounting for 30 % of all isolates which mainly isolated from egg collection center and egg processing plant. ...
Salmonella is a common foodborne pathogen found in poultry production systems. Contaminated poultry products are a major source of human salmonellosis. Understanding the conditions of contamination and the genetic relationships of Salmonella in poultry production is necessary to develop effective interventions measures for controlling Salmonella transmission. Through epidemiological investigation and whole-genome sequence (WGS) analysis, this study revealed the epidemic law, drug resistance, and genetic characteristics of Salmonella in the production chain. In total, 130 (10.77 %) Salmonella strains were isolated from the collected samples, with Salmonella Enteritidis (S. Enteritidis) and S. Infantis being the predominant serotypes. WGS analysis revealed that the Sequence Type (ST) of all the 22 strains of S. Enteritidis were ST11, and 13 strains of S. Infantis were ST32. Antimicrobial resistance gene analysis showed that 95.45 % of S. Enteritidis carried strA/strB, sul, and blaTEM-1B; 77.3 % contained tetA genes; and 100 % of S. Infantis carried aac(6′)-Iaa and mdf(A), which was consistent with the drug resistance phenotype. The phylogenetic tree showed that S. Enteritidis strains with different links were distributed in the same branch which displayed the very close genetic relationship. Combined with epidemiological investigations, it was found that S. Enteritidis infection begins at the chicken breeding stage and spreads vertically and horizontally along the production chain. In addition, phylogenetic tree analysis of S. Infantis showed that the genomes of 13 strains from egg products and egg collection center were very similar and belonged to the same clone cluster. It is speculated that the S. Infantis transmitted in the industrial chain is the same strain; it can spread along the industrial chain, and cross-contamination may occur. This study indicates that Salmonella contamination in the layered industrial chain is more serious and that a cloning relationship exists among the strains of different links. Therefore, more stringent measures should be taken to control Salmonella during the chicken breeding stage, and the importance of implementing good hygiene practices at every level of production should be emphasized.
... Regarding human infections, Salmonella serotypes Enteritidis (SE) and Typhimurium are considered the most important serovars with the greatest impact on public health in several countries (Galán-Relaño et al. 2023). Moreover, poultry products are still the most important source of human infection (Mattock et al. 2024). The first line of treatment for this bacterial illness is antimicrobial therapy; however, due to antibiotic abuse in both human medicine and animal husbandry, antimicrobial resistance has become an issue (Castro-Vargas et al. 2020). ...
Salmonella serotypes Enteritidis (SE) and Typhimurium are thought to be the most significant serovars in terms of human infections, having the most effects on public health. In addition, chicken products continue to be the main source of illness for humans. The global chicken supply is getting increasingly interconnected, and the infection of SE is spreading globally as a result of breeding stocks that are contaminated being traded internationally. In addition to foodborne transmission, cross-contamination in kitchens or food processing areas and intimate contact with animals or habitats that are contaminated can also transmit SE. The poultry industry has been under constant pressure to reduce or stop using antibiotics in production for more than a decade due to concerns about food safety (including antibiotic resistance and consumer expectations) and the actual effects of pathogen contamination on food safety. Essential oils are well-known for being organic, odorous, volatile substances with potential medicinal benefits that have attracted a lot of interest from scientists, particularly in the chicken sector. This chapter explains Salmonella ser. Enteritidis as a unique foodborne pathogen, provides updates on the use of essential oils as natural antibiotic substitutes in the poultry industry, and discusses the use of Lippia origanoides to lessen the effects of SE in broiler chickens. This plant exhibits a wide range of chemotypes, with the majority having distinct essential oil compositions. Furthermore, it produces essential oils with a wide range of chemical compositions, including the so-called chemotypes of thymol and carvacrol. It is an alternate method of controlling SE in poultry production.
... Various reports have indicated the widespread surge of S. Infantis in the poultry sector, especially in broilers, thus possibly being a route for human infections [2,3,15]. From the literature search hereby conducted, a total of 13 articles concerning S. Infantis detection at the broiler farm level were deemed eligible. ...
Salmonella enterica subsp. enterica serovar Infantis poses a growing threat to public health, due to its increasing prevalence worldwide and its association with high levels of antimicrobial resistance. Among livestock, S. Infantis is especially isolated from broilers. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic review was conducted by searching in three databases (Web of Science, Scopus, and PubMed) for English-language studies (1957–2023) that reported the prevalence of S. Infantis in broiler farms. Eligible studies included epidemiological investigations conducted in broiler chickens by sampling the house environment (flock-level prevalence) or the birds (individual-level prevalence). A random-effect model was applied to calculate S. Infantis pooled prevalence estimates with 95% confidence intervals (CIs). Furthermore, to assess between-study heterogeneity, the inconsistency index statistic (I2) was calculated. Among 537 studies retrieved, a total of 9 studies reporting flock-level prevalence of S. Infantis and 4 reporting individual-level prevalence were retained for analysis. The flock-level pooled prevalence was estimated to be 9% (95% CI: 1–26%) and a high between-study heterogeneity was found (I2 = 99%, p < 0.01). Concerning individual-level prevalence, a meta-analysis was not performed due to the scarcity of eligible studies. The data presented underscore the significant occurrence of S. Infantis in broilers at the farm level. By summarizing the existing literature, this work provides useful insights for conducting future surveys of Salmonella spp. in live broiler chickens as a preliminary step for developing more efficient control strategies.
... Salmonella enterica serovar Infantis is an important zoonotic pathogen with rising prevalence and virulence [21][22][23]. While studies have investigated the role of O-antigen in Salmonella virulence [7][8][9], little is known about its regulatory mechanism in S. Infantis virulence. ...
Salmonella enterica serovar Infantis has emerged as a prevalent foodborne pathogen in poultry with significant global health implications. This study investigates the molecular characteristics influencing virulence in a S. Infantis rough variant collected from a poultry farm in the USA. In this study, whole genome sequencing and comparative genomics were performed on smooth and rough poultry S. Infantis isolates, while chicken embryo lethality assay was conducted to assess their virulence. Comparative genomics between isolates was analyzed using Mauve pairwise Locally Collinear Blocks to measure the genetic conservation. Embryo survival rates between the isolates were compared using the Kaplan–Meier curves. High genomic conservation was observed between the two isolates, but a frameshift mutation was detected in the Wzz(fepE) gene of the rough variant, resulting in early protein truncation. The chicken embryo lethality assay showed that the lethality rate of the smooth strain was higher than that of the rough strain (p < 0.05). This study identifies a frameshift mutation in the Wzz(fepE) gene, leading to protein truncation, which may reduce bacterial virulence by impacting O-antigen biosynthesis in the rough Salmonella Infantis variant. These findings deepen our understanding of S. Infantis pathogenesis and suggest that targeting the Wzz(fepE) gene or related pathways could be a promising strategy for developing effective vaccines and therapeutic interventions.
... enterica serovar Infantis (S. Infantis), a globally prevalent serotype associated with foodborne diseases, poses significant public health concerns due to its antimicrobial resistance and ability to form biofilms [1,2]. The antimicrobial resistance of S. Infantis is mainly associated with the presence of the pESI plasmid, which confers resistance to tetracyclines, streptomycin, sulfonamides, fluoroquinolones, and trimethoprim [3,4]. ...
Salmonella Infantis poses a significant challenge in poultry production due to its persistence and resistance to disinfectants. This study investigated the survival of the S. Infantis strain on different surfaces and evaluated the efficacy of disinfectants in both preventing and treating biofilms. The survival of the tested S. Infantis strain was assessed on plastic and stainless steel surfaces after 24 and 48 h. The minimum inhibitory concentrations (MICs) of five disinfectants were determined, and their antiadhesion effectiveness was evaluated using crystal violet. The efficacy of biofilm treatment was evaluated by cell culturability. The results showed that the adhesion of S. Infantis was significantly higher on the plastic surface. The disinfectants were effective at reducing biofilm formation only within the first 24 h. Fresh solutions of disinfectants based on quaternary ammonium compounds exhibited the highest antimicrobial efficacy, while chlorocresol was the most effective for both the prevention and treatment of biofilms. The study results suggest that the presence of plastic surfaces may contribute to the dissemination of Salmonella. Additionally, the effectiveness of disinfectants varied based on storage conditions and contact time, while biofilms demonstrated reduced susceptibility compared to planktonic cells. However, given the laboratory scale of this study, further validation on a commercial scale is necessary to confirm these findings.
... Interestingly, all S. Infantis isolates, except only one, from the whole carcass washings were sensitive to the selected antimicrobial in the present study, which was resistant to nalidixic acid. CDC is more concerned about S. Infantis, as an emerging human multidrug-resistant pathogen [74]. Antimicrobials are used more extensively in Salmonella than Campylobacter as it causes clinical infection in poultry. ...
The persistence of non-typhoidal Salmonella and Campylobacter in chicken meat is a considerable public health risk and a future challenge. This study aimed to determine the prevalence of Salmonella and Campylobacter in poultry processing lines where different chlorine concentrations were used in the chill tank. The samples were collected from four types of processing plants in Sri Lanka, considering the chlorine concentration used in the chill tank, which ranged from 2 ppm to 50 ppm. Salmonella and Campylobacter were isolated from whole carcass washings, neck skin, and cecal samples. Subsequently, an antimicrobial susceptibility test was performed for the isolates. The results revealed the overall prevalence of Salmonella and Campylobacter was 78.25% and 63.5%, respectively. Positive percentages of Salmonella and Campylobacter were high in the carcasses compared to the neck skin and ceca. The Campylobacter counts on the whole carcasses were significantly low (p < 0.001), at higher chlorine concentrations ranging from 20 to 30 ppm and 40 to 50 ppm. The pathogen prevalence in the whole carcasses was 84.7% Campylobacter coli, 39.1% Campylobacter jejuni, 71.1% Salmonella Typhimurium, and 28.8% Salmonella Infantis. The highest resistance was observed for tetracycline (63.8%) in Salmonella, while it was for gentamicin (87.8%) in Campylobacter. The prevalence percentage of multidrug-resistant Campylobacter was 51.2%, while it was 2.12% for Salmonella. The persistence of multidrug-resistant Salmonella and Campylobacter on the post-chill carcasses was highlighted in the present study as a significant public health threat that has to be addressed urgently.
Contemporary challenges of European food law include ensuring food compliance with specific microbiological criteria. EU law’s focus on Salmonella is based on its two serotypes, i.e. typhimurium and enteritidis. Therefore, the detection of other serotypes such as kentucky, newport or infantis in fresh poultry meat or a flock of laying hens can lead to fresh meat or eggs being recognized as compliant with the criteria, thereby placing them on the market. There is also a question about other non-regulated poultry species, such as ducks and geese, as well as meat and eggs obtained from them. The CJEU judgment of 28 April 2022 in case C-89/21 stated that national authorities should not have limited ability to detect and calculate other microorganisms, and EU rules should be interpreted in such a way that the competent veterinary public authority may also consider as dangerous other categories of foodstuffs in which various other pathogenic microorganisms have been detected. So the question arises: can the Veterinary Inspection order the withdrawal from the market of products of animal origin contaminated with Salmonella or eggs obtained from poultry contaminated with this bacteria, despite – at least apparent and superficial – compliance with the EU regulations?
Non-typhoidal Salmonella enterica is a leading cause of gastrointestinal illnesses in the United States. Among the 2,600 different S. enterica serovars, Infantis has been significantly linked to human illnesses and is frequently recovered from broilers and chicken parts in the U.S. A key virulence determinant in serovar Infantis is the presence of the megaplasmid pESI, conferring multidrug resistance. To further characterize the virulence potential of this serovar, the present study identified the types of plasmids harbored by Infantis strains, recovered from surface waters adjacent to leafy greens farms in California. Sequencing analysis showed that each of the examined 12 Infantis strains had a large plasmid ranging in size from 78 kb to 125 kb. In addition, a second 4-kb plasmid was detected in two strains. Plasmid nucleotide queries did not identify the emerging megaplasmid pESI in the examined Infantis strains; however, the detected plasmids each had similarity to a plasmid sequence already cataloged in the nucleotide databases. Subsequent comparative analyses, based on gene presence or absence, divided the detected plasmids into five distinct clusters, and the phylogram revealed these Infantis plasmids were clustered based either on the plasmid conjugation system, IncI and IncF, or on the presence of plasmid phage genes. Assignment of the putative genes to functional categories revealed that the large plasmids contained genes implicated in cell cycle control and division, replication and recombination and defense mechanisms. Further analysis of the mobilome, including prophages and transposons, demonstrated the presence of genes implicated in the release of the bactericidal peptide microcin in the IncF plasmids and identified a Tn10 transposon conferring tetracycline resistance in one of the IncI1 plasmids. These findings indicated that the plasmids in the environmental S. enterica serovar Infantis strains from surface waters harbored a wide variety of genes associated with adaptation, survivability and antimicrobial resistance.
Poultry can act as a reservoir of Salmonella enterica serotype Infantis (S. Infantis) of clinical and epidemiological importance because it triggers foodborne disease outbreaks and presents antibiotic multiresistance. The present study aimed to determine antimicrobial resistance in S. Infantis isolates from poultry livers, litter and chicken carcasses from Lima, Peru, during 2022 - 2023. S. Infantis was isolated from 10.1% (27/267), 4.7% (4/86), and 8% (2/25) of the bird, litter and carcass samples, respectively. All isolates showed resistance to nalidixic acid (NA). In addition, 94%, 76%, 70%, 48%, and 45% of the isolates showed resistance to tetracycline (TET), amoxicillin (AMX), chloramphenicol (C), gentamicin (GEN) and trimethoprim/sulfamethoxazole (TS), respectively. All isolates showed susceptibility to imipinem and amoxicillin/clavulanic acid. Of the isolates, 93.9% displayed multidrug resistance. The most frequent resistance pattern was C-AMX-NA-GEN-TET (24.2%, n=8), determined even from carcass isolates, followed by C-AMX-NA-TET (18.2%, n=6), and C-AMX-NA-GEN-TS-TET (12.1%, n=4). The presence of these multidrug-resistant S. Infantis isolates is a threat to food safety and public health.
Salmonella Infantis is one of the most frequent serovars reported in broilers and is also regularly identified in human salmonellosis cases, representing a relevant public health problem. In the laboratories of the Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata (IZSPB), six Salmonella Infantis strains with antigenic formula-:r:1,5 have been isolated from the litter and carcass of broilers between 2018 and 2022. The strains were investigated to evaluate their phenotype, antibiotic resistance and genomic profiles. Genomic analysis confirmed that the isolates belonged to the Infantis serotype and to the sequence type ST32. Moreover, all strains showed a multidrug-resistant (MDR) profile and were characterised by the presence of the IncFIB plasmid incompatibility group. Three strains had the bla CTX-M-1 gene, and one of them carried IncX1. The presence of this new variant of S. Infantis is particularly relevant because it could expand the landscape of the S. Infantis population. The absence of the somatic antigen could pose a problem in both isolation and serotyping and a consequent public health concern due to the spread of Salmonella infection.
Salmonella enterica subsp. enterica serovar Infantis is the most prevalent serovar found in broilers and broiler meat and is among the top five serovars responsible for human infections in Europe. In 2008, a multidrug-resistant S. Infantis isolate emerged in Israel with a mosaic megaplasmid named pESI, associated with increased virulence, biofilm formation, and multidrug resistance. Since then, S. Infantis clones with pESI-like plasmids have been reported worldwide, replacing pESI-free clones. Here, we typed 161 S. Infantis isolates of poultry (n = 133) and human clinical (n = 28) origin using whole-genome sequencing. The isolates were collected between 2007 and 2021. In addition, we performed PacBio/Illumina sequencing for two representative pESI-like plasmids and compared them with publicly available sequences. All isolates belonged to sequence type 32 (ST32), except for one isolate that represented a novel single-locus variant of ST32. Core genome MLST (cgMLST) analysis revealed 14 clusters of genetically closely related isolates, of which four suggested broiler-to-human transmission of S. Infantis. pESI-like plasmids were present in 148/161 (91.9%) isolates; all were highly similar to the publicly available pESI-like sequences but lacked extended-spectrum beta-lactamase (ESBL) genes. PacBio/Illumina hybrid assembly allowed the reconstruction of two novel complete pESI variants. The present study revealed that the multidrug-resistant, pESI-positive S. Infantis clone became the predominant S. Infantis clone in Slovenian broilers and humans during the last decade. Continued surveillance of resistant S. Infantis clones along the food chain is needed to guide public health efforts. IMPORTANCE Salmonella Infantis clones with pESI-like plasmids harboring several virulence and resistance genes have been reported worldwide. In the present study, we compared the population structure of 161 Salmonella Infantis isolates obtained from humans and broilers in Slovenia from 2007 to 2021. Whole-genome sequencing showed that most human isolates clustered apart from broiler isolates, suggesting an alternative source of infection. Most isolates were multidrug resistant due to the presence of pESI-like plasmids, of which two variants (pS89 and pS19) were fully reconstructed using long-read sequencing. Both exhibited high similarity with the original Israeli pESI plasmid and German p2747 plasmid. The prototype plasmid pS89 harbored the typical pESI-associated resistance genes aadA1, qacEΔ1, sul1, and tet(A), which were absent in the truncated plasmid pS19.
Infantis has recently become one of the most common serotypes of Salmonella isolated in the U.S. from raw meat samples collected in processing facilities and in retail stores. Investigations have determined that the majority of these isolates contain the pESI plasmid, but there has not been a large-scale investigation of the chromosome of these isolates. Here, we investigated 3276 whole-genome sequences of Salmonella Infantis with and without the pESI plasmid to understand chromosomal differences between plasmid carriage groups. S. Infantis genomes arranged into multiple clades with a single clade containing the isolates carrying the plasmid. Fifty-eight SNPs were identified in complete linkage disequilibrium between isolates that did and did not carry the plasmid. However, there were no unique genes present only in the genomes of isolates containing the plasmid. On average, isolates with the plasmid did contain more insertion sequences than those without (p < 0.05). Given that S. Infantis isolates carrying pESI form a single clade, it can be inferred that the increase in carriage of this plasmid in the U.S. is due to rapid clonal expansion of a single strain rather than as a result of multiple transfer events. As this S. Infantis clone does not contain any unique chromosomal genes, its proliferation appears to be due to pESI plasmid-encoded genes that may be advantageous in the chickens and turkeys or in their environment.
Salmonella Infantis is presenting an increasing risk to public health. Of particular concern are the reports of pESI, a multidrug resistance (MDR) encoding megaplasmid, in isolates from multiple countries, but little is known about its presence or diversity in South Africa. Whole genome sequences of 387 S . Infantis isolates from South Africa (2004–2020) were analysed for genetic phylogeny, recombination frequency, antimicrobial resistance (AMR) determinants, plasmid presence and overall gene content. The population structure of South African S . Infantis was substantially different to S . Infantis reported elsewhere; only two thirds of isolates belonged to eBG31, while the remainder were identified as eBG297, a much rarer group globally. Significantly higher levels of recombination were observed in the eBG297 isolates, which was associated with the presence of prophages. The majority of isolates were putatively susceptible to antimicrobials (335/387) and lacked any plasmids (311/387); the megaplasmid pESI was present in just one isolate. A larger proportion of eBG31 isolates, 19% (49/263), contained at least one AMR determinant, compared to eBG297 at 2% (3/124). Comparison of the pan-genomes of isolates from either eBG identified 943 genes significantly associated with eBG, with 43 found exclusively in eBG31 isolates and 34 in eBG297 isolates. This, along with the single nucleotide polymorphism distance and difference in resistance profiles, suggests that eBG31 and eBG297 isolates occupy different niches within South Africa. If antibiotic-resistant S . Infantis emerges in South Africa, probably through the spread of the pESI plasmid, treatment of this infection would be compromised.
Salmonella enterica serovar Infantis is one of the five main causes of human salmonellosis in the European Union (EU) and in recent years, has been increasingly reported to carry multiple antimicrobial resistance determinants, including extended-spectrum beta-lactamase (ESBL) genes. In our study, we used WGS-based tools to characterize S. Infantis strains circulating in the Abruzzo and Molise regions of Italy between 2017 and 2020 and compared this local dataset to the S. Infantis population present in Italy over the last two decades. Phylogenetic analyses demonstrated that the majority of strains isolated from poultry and turkeys from Abruzzo and Molise were closely related and belonged to one of the two main genetic clusters present in Italy, which were grouped predominantly as ESBL-producing strains that harbored pESI-like plasmid. We showed that 60% of the local strains carried multiple antibiotic resistance genes, including ESBL gene bla CTX–M–1 as well as aad A1, dfr A1, dfr A14, sul1 , and tet (A) genes present on the pESI-like megaplasmid. The analysis of strains from Abruzzo and Molise and the publicly available Italian S. Infantis sequences revealed a dramatic increase in the number of identified AMR genes in the strains isolated after 2011. Moreover, the number of strains resistant to five or more antibiotic classes increased from 20–80% in the last decade likely due to the acquisition of the megaplasmid. The persistence of the ESBL-producing and the multidrug-resistant (MDR) clone of S. Infantis in poultry populations in Italy and in Europe requires rapid and efficient intervention strategies to prevent further expansion of the clone.
The poultry industry in Iran is the main supplier of protein in the food chain. In the present study, we showed the importance of the possible dissemination of clonally related multiple drug resistant (MDR) Salmonella Infantis in broiler farms in Iran. In total, 156 fecal samples belonging to 23 poultry farms in Razavi Khorasan province, northeast of Iran, were examined for the presence of Salmonella serovars. Molecular serotypes and serogroups, class 1 and 2 integron types, colistin resistance genes ( mcr1 and mcr2) and antimicrobial susceptibility patterns were determined on the recovered Salmonella isolates. Based on PCR analysis, 30 recovered Salmonella isolates were identified asS. Infantis (23 isolates; 76.6%),S. Enteritidis
(six isolates; 20%), and one isolate (3.3%) was not serotyped by the applied method. Class 1 integrons were detected in 22 isolates (95.6%) and class 2 integrons were not detected in any of the isolates. Although colistin resistance was prevalent in disc diffusion test, mcr-1 and mcr-2 genes were not detected. All class 1 integrons carried the cassette aadA1 gene. All Salmonella isolates were resistant to colistin and amoxicillin/clavulanic acid and MDR patterns were observed in most (96.6%) isolates. This study revealed a high prevalence rate of S. Infantis and the presence of class 1 integrons in broiler farms. The presence of the same integron cassettes in the sequenced isolates suggests that strains are clonally related. Stringent monitoring programs are required to prevent the spreading of MDR Salmonella serovars into food chain via poultry products.
Salmonella comprises over 2500 serotypes and foodborne contamination associated with this pathogen remains an important health concern worldwide. During the last decade, a shift in serotype prevalence has occurred as traditionally less prevalent serotypes are increasing in frequency of infections, especially those related to poultry meat contamination. S. Infantis is one of the major emerging serotypes, and these strains commonly display antimicrobial resistance and can persist despite cleaning protocols. Thus, this work aimed to isolate S. Infantis strains from a poultry meat farm in Santiago, Chile and to characterize genetic variations present in them. We determined their genomic and phenotypic profiles at different points along the production line. The results indicate that the strains encompass 853 polymorphic sites (core-SNPs) with isolates differing from one another by 0–347 core SNPs, suggesting variation among them; however, we found discrete correlations with the source of the sample in the production line. Furthermore, the pan-genome was composed of 4854 total gene clusters of which 2618 (53.9%) corresponds to the core-genome and only 181 (3.7%) are unique genes (those present in one particular strain). This preliminary analysis will enrich the surveillance of Salmonella, yet further studies are required to assess their evolution and phylogeny.
Salmonella enterica serovar Infantis is the fifth most common Salmonella serovar isolated in England and Wales. Epidemiological, genotyping and antimicrobial-resistance data for S . enterica Infantis isolates were used to analyse English and Welsh demographics over a 5 year period. Travel cases associated with S . enterica Infantis were mainly from Asia, followed by cases from Europe and North America. Since 2000, increasing numbers of S . enterica Infantis had multidrug resistance determinants harboured on a large plasmid termed ‘plasmid of emerging S . enterica Infantis’ (pESI). Between 2013 and 2018, 42 S . enterica Infantis isolates were isolated from humans and food that harboured resistance determinants to multiple antimicrobial classes present on a pESI-like plasmid, including extended-spectrum β-lactamases (ESBLs; bla CTX-M-65 ). Nanopore sequencing of an ESBL-producing human S . enterica Infantis isolate indicated the presence of two regions on an IncFIB pESI-like plasmid harbouring multiple resistance genes. Phylogenetic analysis of the English and Welsh S . enterica Infantis population indicated that the majority of multidrug-resistant isolates harbouring the pESI-like plasmid belonged to a single clade maintained within the population. The bla CTX-M-65 ESBL isolates first isolated in 2013 comprise a lineage within this clade, which was mainly associated with South America. Our data, therefore, show the emergence of a stable resistant clone that has been in circulation for some time in the human population in England and Wales, highlighting the necessity of monitoring resistance in this serovar.
Food-borne infections with Salmonella are among the most common causes of human diseases worldwide, and infections with the serovar Infantis are becoming increasingly important. So far, diverse phenotypes and genotypes of S. Infantis have been reported. Therefore, the present study aimed to investigate the infection dynamics of two different S. Infantis strains in broilers. For this purpose, 15 birds were infected on day 2 of life with 108 CFU/ml of a pESI+ or a pESI- S. Infantis strain, respectively. Ten uninfected birds served as in-contact birds to monitor transmission. In both groups, an increase of infection was observed from 7 days of age onwards reaching its peak at 28 days. However, the pESI+ strain proved significantly more virulent being re-isolated from most cloacal swabs and organs by direct plating. In contrast, the pESI- strain could be re-isolated from cloacal swabs and caeca only when enrichment was applied. Although the excretion of this strain was limited, the transmission level to in-contact birds was similar to the pESI+ strain. Differences in infection dynamics were also reflected in the antibody response: whereas the pESI+ strain provoked a significant increase in antibodies, antibody levels following infection with the pESI- strain remained in the range of negative control birds. The actual findings provide for the first time evidence of S. Infantis strain-specific virulence in broilers and confirm previous observations in the field regarding differences in persistence on farms and resistance against disinfectants.
Salmonella Infantis is a poultry-adapted Salmonella enterica serovar that is increasingly reported in broilers and is also regularly identified among human salmonellosis cases. An emerging S. Infantis mega-plasmid (pESI), carrying fitness, virulence and antimicrobial resistance genes, is also increasingly found. We investigated the prevalence, genetic characteristics and risk factors for (pESI-carrying) S. Infantis in broilers. Faecal samples from 379 broiler flocks (in 198 farms with ≥3000 birds) in the Netherlands were tested. A questionnaire about farm characteristics was also administered. Sampling was performed in July 2018-May 2019, three weeks before slaughter. Fourteen flocks (in 10 farms) were S. Infantis-positive, resulting in a 3.7% flock-level and 5.1% farm-level prevalence. Based on multi-locus sequence typing (MLST), all isolates belonged to sequence type 32. All but one isolate carried a pESI-like mega-plasmid. Core-genome MLST showed considerable heterogeneity among the isolates, even within the same farm, with a few small clusters detected. The typical pESI-borne multi-resistance pattern to aminoglycosides, sulphonamide and tetracycline (93%), as well as trimethoprim (71%), was found. Additionally, resistance to (fluoro)quinolones based on gyrA gene mutations was detected. S. Infantis was found more often in flocks using salinomycin as coccidiostat, where flock thinning was applied or litter quality was poor, whereas employing external cleaning companies, wheat in feed, and vaccination against infectious bronchitis, were protective. Suggestive evidence for vertical transmission from hatcheries was found. A heterogeneous (pESI-carrying) S. Infantis population has established itself in Dutch broiler flocks, calling for further monitoring of its spread and a comprehensive appraisal of control options.
Molecular clock models relate observed genetic diversity to calendar time, enabling estimation of times of common ancestry. Many large datasets of fast-evolving viruses are not well fitted by molecular clock models that assume a constant substitution rate through time, and more flexible relaxed clock models are required for robust inference of rates and dates. Estimation of relaxed molecular clocks using Bayesian Markov chain Monte Carlo is computationally expensive and may not scale well to large datasets. We build on recent advances in maximum likelihood and least-squares phylogenetic and molecular clock dating methods to develop a fast relaxed-clock method based on a Gamma-Poisson mixture model of substitution rates. This method estimates a distinct substitution rate for every lineage in the phylogeny while being scalable to large phylogenies. Unknown lineage sample dates can be estimated as well as unknown root position. We estimate confidence intervals for rates, dates, and tip dates using parametric and non-parametric bootstrap approaches. This method is implemented as an open-source R package, treedater.
Salmonella enterica is one of the most important foodborne pathogens around the world. In the last years, S. enterica serovar Infantis has become an important emerging pathogen in many countries, often as multidrug resistant clones. To understand the importance of S. enterica in the broiler industry in Ecuador, we performed a study based on phenotypic and WGS data of isolates from poultry farms, chicken carcasses and humans. We showed a high prevalence of S. enterica in poultry farms (41.4%) and chicken carcasses (55.5%), but a low prevalence (1.98%) in human samples. S. Infantis was shown to be the most prevalent serovar with a 98.2, 97.8, and 50% in farms, foods, and humans, respectively, presenting multidrug resistant patterns. All sequenced S. Infantis isolates belonged to ST32. For the first time, a pESI-related megaplasmid was identified in Ecuadorian samples. This plasmid contains genes of antimicrobial resistance, virulence factors, and environmental stress tolerance. Genomic analysis showed a low divergence of S. Infantis strains in the three analyzed components. The results from this study provide important information about genetic elements that may help understand the molecular epidemiology of S. Infantis in Ecuador.
This report of the European Food Safety Authority and the European Centre for Disease Prevention and Control presents the results of zoonoses monitoring activities carried out in 2018 in 36 European countries (28 Member States (MS) and 8 non-MS). The first and second most commonly reported zoonoses in humans were campylobacteriosis and salmonellosis, respectively. The European Union (EU) trend for confirmed human cases of these two diseases was stable during 2014-2018. The proportion of human salmonellosis cases due to Salmonella Enteritidis was at the same level in 2018 as in 2017. Of the 27 reporting MS, 16 met all Salmonella reduction targets for poultry, whereas 11 MS failed meeting at least one. The EU flock prevalence of target Salmonella serovars in breeding hens, laying hens, broilers and fattening turkeys decreased during recent years but stalled in breeding turkeys. Salmonella results from Competent Authorities for pig carcasses and for poultry tested through National Control Programmes were more frequently positive compared with food business operators. Shiga toxin-producing Escherichia coli (STEC) infections in humans were the third most commonly reported zoonosis in the EU and increased from 2014 to 2018. Yersiniosis was the fourth most frequently reported zoonosis in humans in 2018 with a stable trend in 2014-2018. The number of reported confirmed listeriosis cases further increased in 2018, despite Listeria rarely exceeding the EU food safety limit tested in ready-to-eat food. In total, 5,146 food- and waterborne outbreaks were reported. Salmonella was the most commonly detected agent with S. Enteritidis causing one in five outbreaks. Salmonella in eggs and egg products was the highest risk agent/food pair. A large increase of human West Nile virus infections was reported in 2018. The report further updates on bovine tuberculosis, Brucella, Trichinella, Echinococcus, Toxoplasma, rabies, Coxiella burnetii (Q fever) and tularaemia.
Salmonella Infantis is one of the five serovars most frequently causing human salmonellosis in Europe, mainly associated with poultry. A clone harbouring a conjugative plasmid of emerging S. Infantis (pESI)-like megaplasmid, carrying multidrug resistant (MDR) and extended-spectrum beta-lactamases (ESBL) genes, has spread in the Italian broiler chicken industry also causing human illness. This work is aimed at elucidating the molecular epidemiology of S. Infantis and pESI-like in Europe using whole-genome sequencing and bioinformatics analysis, and to investigate the genetic relatedness of S. Infantis clones and pESI-like from animals, meat, feed and humans provided by institutions of nine European countries. Two genotyping approaches were used: chromosome or plasmid SNP-based analysis and the minimum spanning tree (MST) algorithm based on core-genome multilocus sequence typing (cgMLST). The European S. Infantis population appeared heterogeneous, with different genetic clusters defined at core-genome level. However, pESI-like variants present in 64.1 % of the isolates were more genetically homogeneous and capable of infecting different clonal lineages in most of the countries. Two different pESI-like with ESBL genes (n=82) were observed: blaCTX-M-1-positive in European isolates and blaCTX-M-65-positive in American isolates (study outgroup). Both variants had toxin-antitoxin systems, resistance genes towards tetracyclines, trimethoprim, sulphonamides and aminoglycosides, heavy metals (merA) and disinfectants (qacEΔ). Worryingly, 66 % of the total isolates studied presented different gyrA chromosomal point mutations associated with (fluoro)quinolone resistance (MIC range 0.125-0.5 mg/L), while 18 % displayed transferable macrolide resistance mediated by mph, mef and erm(B) genes. Proper intervention strategies are needed to prevent further dissemination/transmission of MDR S. Infantis and pESI-like along the food chain in Europe.
Background:
Salmonella Infantis (S. Infantis) is one of the most frequent Salmonella serovars isolated from human cases of salmonellosis and the most detected serovar from animal and food sources in Europe. The serovar is commonly associated with poultry and there is increasing concern over multidrug resistant clones spreading worldwide, as the dominating clones are characterized by presence of large plasmids carrying multiple resistance genes. Increasing the knowledge of the S. Infantis population and evolution is important for understanding and preventing further spread. In this study, we analysed a collection of strains representing different decades, sources and geographic locations. We analysed the population structure and the accessory genome, in particular we identified prophages with a view to understand the role of prophages in relation to the evolution of this serovar.
Results:
We sequenced a global collection of 100 S. Infantis strains. A core-genome SNP analysis separated five strains in e-Burst Group (eBG) 297 with a long branch. The remaining strains, all in eBG31, were divided into three lineages that were estimated to have separated approximately 150 years ago. One lineage contained the vast majority of strains. In five of six clusters, no obvious correlation with source or geographical locations was seen. However, one cluster contained mostly strains from human and avian sources, indicating a clone with preference for these sources. The majority of strains within this cluster harboured a pESI-like plasmid with multiple resistance genes. Another lineage contained three genetic clusters with more rarely isolated strains of mainly animal origin, possibly less sampled or less infectious clones. Conserved prophages were identified in all strains, likely representing bacteriophages which integrated into the chromosome of a common ancestor to S. Infantis. We also saw that some prophages were specific to clusters and were probably introduced when the clusters were formed.
Conclusions:
This study analysed a global S. Infantis population and described its genetic structure. We hypothesize that the population has evolved in three separate lineages, with one more successfully emerging lineage. We furthermore detected conserved prophages present in the entire population and cluster specific prophages, which probably shaped the population structure.
We present fastbaps, a fast solution to the genetic clustering problem. Fastbaps rapidly identifies an approximate fit to a Dirichlet process mixture model (DPM) for clustering multilocus genotype data. Our efficient model-based clustering approach is able to cluster datasets 10-100 times larger than the existing model-based methods, which we demonstrate by analyzing an alignment of over 110 000 sequences of HIV-1 pol genes. We also provide a method for rapidly partitioning an existing hierarchy in order to maximize the DPM model marginal likelihood, allowing us to split phylogenetic trees into clades and subclades using a population genomic model. Extensive tests on simulated data as well as a diverse set of real bacterial and viral datasets show that fastbaps provides comparable or improved solutions to previous model-based methods, while being significantly faster. The method is made freely available under an open source MIT licence as an easy to use R package at https://github.com/gtonkinhill/fastbaps.
For many decades, Salmonella enterica has been subdivided by serological properties into serovars or further subdivided for epidemiological tracing by a variety of diagnostic tests with higher resolution. Recently, it has been proposed that so-called eBurst groups (eBGs) based on the alleles of seven housekeeping genes (legacy multilocus sequence typing [MLST]) corresponded to natural populations and could replace serotyping. However, this approach lacks the resolution needed for epidemiological tracing and the existence of natural populations had not been independently validated by independent criteria. Here, we describe EnteroBase, a web-based platform that assembles draft genomes from Illumina short reads in the public domain or that are uploaded by users. EnteroBase implements legacy MLST as well as ribosomal gene MLST (rMLST), core genome MLST (cgMLST), and whole genome MLST (wgMLST) and currently contains over 100,000 assembled genomes from Salmonella. It also provides graphical tools for visual interrogation of these genotypes and those based on core single nucleotide polymorphisms (SNPs). eBGs based on legacy MLST are largely consistent with eBGs based on rMLST, thus demonstrating that these correspond to natural populations. rMLST also facilitated the selection of representative genotypes for SNP analyses of the entire breadth of diversity within Salmonella. In contrast, cgMLST provides the resolution needed for epidemiological investigations. These observations show that genomic genotyping, with the assistance of EnteroBase, can be applied at all levels of diversity within the Salmonella genus.
Antimicrobial resistance (AMR) is one of the major threats to human and animal health worldwide, yet few high-throughput tools exist to analyse and predict the resistance of a bacterial isolate from sequencing data. Here we present a new tool, ARIBA, that identifies AMR-associated genes and single nucleotide polymorphisms directly from short reads, and generates detailed and customizable output. The accuracy and advantages of ARIBA over other tools are demonstrated on three datasets from Gram-positive and Gram-negative bacteria, with ARIBA outperforming existing methods.
We report a 4.99-Mb draft genome sequence of Salmonella enterica subsp. enterica serovar Infantis strain SPE101, isolated from feces of a 5-month-old breast-fed female showing diarrhea associated with severe dehydration and malnutrition. The infection prolonged for 6 months despite antibiotic treatment.
Objectives: The aim of this study was to characterize a collection of 520 Salmonella enterica serovar Infantis strains isolated from food (poultry meat), human infections and environmental sources from the years 2010, 2013 and 2015 in Switzerland.
Methods: We performed antimicrobial susceptibility testing and pulsed-field gel electrophoresis (PFGE) analysis on all 520 S. Infantis isolates, and whole genome sequencing (WGS) on 32 selected isolates.
Results: The majority (74.8%) of the isolates was multidrug resistant (MDR). PFGE analysis revealed that 270 (51.9%) isolates shared an identity of 90%. All isolates subjected to WGS belonged to sequence type (ST) 32 or a double-locus variant thereof (one isolate). Seven (21.9%) of the sequenced isolates were phylogenetically related to the broiler-associated clone B that emerged in Hungary and subsequently spread within and outside of Europe. In addition, three isolates harboring blaCTX-M-65 on a predicted large (∼320 kb) plasmid grouped in a distinct cluster.
Conclusion: This study documents the presence of the Hungarian clone B and related clones in food and human isolates between 2010 and 2015, and the emergence of a blaCTX-M-65 harboring MDR S. serovar Infantis lineage.
We sequenced the genomes of ten Salmonella enterica serovar Infantis containing blaCTX-M-65 isolated from chicken, cattle, and human sources collected between 2012 and 2015 in the United States through routine NARMS surveillance and product sampling programs. We also completely assembled the plasmids from four of the isolates. All isolates had a D87Y mutation in the gyrA gene and harbored between 7 and 10 resistance genes ( aph (4)-Ia, aac (3)-IVa, aph(3’ )-Ic, blaCTX-M-65, fosA3, floR, dfrA14, sul1, tetA, aadA1 ) located in two distinct sites of a megaplasmid (∼316-323kb) similar to that described in a blaCTX-M-65 -positive S. Infantis isolated from a patient in Italy. High-quality single nucleotide polymorphism (hqSNP) analysis revealed that all U.S. isolates were closely related, separated by only 1 to 38 pairwise high quality SNPs, indicating a high likelihood that strains from humans, chicken, and cattle recently evolved from a common ancestor. The U.S. isolates were genetically similar to the blaCTX-M-65 -positive S. Infantis isolate from Italy, with a separation of 34 to 47 SNPs. This is the first report of the blaCTX-M-65 gene and the pESI-like megaplasmid from S. Infantis in the United States, and illustrates the importance of applying a global One Health, human and animal perspective to combat antimicrobial resistance.
Salmonella is frequently found in poultry and represent an important source for human gastrointestinal infections worldwide. The aim of this study was to investigate the prevalence, genotypes and antimicrobial resistance of Salmonella serotypes in broilers from Ecuador. Caeca content from 388 at random selected broiler batches were collected in 6 slaughterhouses during 1 year and analyzed by the ISO 6579/Amd1 protocol for the isolation for Salmonella. Isolates were serotyped and genotypic variation was acceded by pulsed field gel electrophoresis. MIC values for sulfamethoxazole, gentamicin, ciprofloxacin, ampicillin, cefotaxime, ceftazidime, tetracycline, streptomycin, trimethropim, chloramphenicol, colistin, florfenicol, kanamycin and nalidixic acid were obtained. Presence of blaCTX-M, blaTEM, blaSHV and blaCMY; and mcr-1 plasmid genes was investigated in resistant strains to cefotaxime and colistin respectively. Prevalence at batch level was 16.0%. The most common serotype was S. Infantis (83.9%) followed by S. Enteritidis (14.5%) and S. Corvallis (1.6%). The pulsed field gel electrophoresis analysis showed that S. Corvallis, S. Enteritidis and S. Infantis isolates belonged to 1, 2 and 12 genotypes respectively. S. Infantis isolates showed high resistance rates to 12 antibiotics ranging from 57.7% (kanamycin) up to 98.1% (nalidixic acid and sulfamethoxazole). All S. Enteritidis isolates showed resistance to colistin. High multiresistant patterns were found for all the serotypes. The blaCTX-M gene was present in 33 S. Infantis isolates while mcr-1 was negative in 10 colistin resistant isolates. This study provides the first set of scientific data on prevalence and multidrug-resistant Salmonella coming from commercial poultry in Ecuador.
We report the spread of a clone of multidrug-resistant (MDR), ESBL-producing (blaCTX-M-1) Salmonella enterica subsp. enterica serovar Infantis, in the Italian broiler chicken industry and along the food-chain. This was first detected in Italy in 2011 and led to human infection in Italy in 2013-2014.A set (n = 49) of extended-spectrum cephalosporin (ESC)-resistant (R) isolates of S. Infantis (2011-2014) from humans, food-producing animals and meat thereof, were studied along with a selected set of earlier and more recent ESC-susceptible (ESC-S) isolates (n = 42, 2001-2014). They were characterized by macrorestriction-PFGE analysis and genetic environment of ESC-resistance. Isolates representative of PFGE-patterns and origin were submitted to Whole Genome Sequencing. The emerging ESC-R clone, detected mainly from broiler chickens, broiler meat and humans, showed a minimum pattern of clinical resistance to cefotaxime, tetracycline, sulfonamides, and trimethoprim, beside ciprofloxacin microbiological resistance (MIC 0.25 mg/L). All isolates of this clone harbored a conjugative megaplasmid (~ 280-320 Kb), similar to that described in ESC-susceptible S. Infantis in Israel (pESI-like) in 2014. This megaplasmid carried the ESBL gene blaCTX-M-1, and additional genes [tet(A), sul1, dfrA1 and dfrA14] mediating cefotaxime, tetracycline, sulfonamide, and trimethoprim resistance. It also contained genes conferring enhanced colonization capability, virulence (fimbriae, yersiniabactin), resistance and fitness (qacE1, mer) in the intensive-farming environment. This emerging clone of S. Infantis has been causing infections in humans, most likely through the broiler industry. Since S. Infantis is among major serovars causing human infections in Europe and is an emerging non-typhoidal Salmonella globally, further spread of this lineage in primary productions deserves quick and thorough risk-management strategies.
Illness and death from diseases caused by contaminated food are a constant threat to public health and a significant impediment to socio-economic development worldwide. To measure the global and regional burden of foodborne disease (FBD), the World Health Organization (WHO) established the Foodborne Disease Burden Epidemiology Reference Group (FERG), which here reports their first estimates of the incidence, mortality, and disease burden due to 31 foodborne hazards. We find that the global burden of FBD is comparable to those of the major infectious diseases, HIV/AIDS, malaria and tuberculosis. The most frequent causes of foodborne illness were diarrheal disease agents, particularly norovirus and Campylobacter spp. Diarrheal disease agents, especially non-typhoidal Salmonella enterica, were also responsible for the majority of deaths due to FBD. Other major causes of FBD deaths were Salmonella Typhi, Taenia solium and hepatitis A virus. The global burden of FBD caused by the 31 hazards in 2010 was 33 million Disability Adjusted Life Years (DALYs); children under five years old bore 40% of this burden. The 14 subregions, defined on the basis of child and adult mortality, had considerably different burdens of FBD, with the greatest falling on the subregions in Africa, followed by the subregions in South-East Asia and the Eastern Mediterranean D subregion. Some hazards, such as non-typhoidal S. enterica, were important causes of FBD in all regions of the world, whereas others, such as certain parasitic helminths, were highly localised. Thus, the burden of FBD is borne particularly by children under five years old-although they represent only 9% of the global population-and people living in low-income regions of the world. These estimates are conservative, i.e., underestimates rather than overestimates; further studies are needed to address the data gaps and limitations of the study. Nevertheless, all stakeholders can contribute to improvements in food safety throughout the food chain by incorporating these estimates into policy development at national and international levels.
In the work presented here, we designed and developed two easy-to-use Web tools for in silico detection and characterization of whole-genome sequence (WGS) and whole-plasmid sequence data from members of the family
Enterobacteriaceae. These tools will facilitate bacterial typing based on draft genomes of multidrug-resistant Enterobacteriaceae species by the rapid detection of known plasmid types. Replicon sequences from 559 fully sequenced plasmids associated with
the family Enterobacteriaceae in the NCBI nucleotide database were collected to build a consensus database for integration into a Web tool called PlasmidFinder
that can be used for replicon sequence analysis of raw, contig group, or completely assembled and closed plasmid sequencing
data. The PlasmidFinder database currently consists of 116 replicon sequences that match with at least at 80% nucleotide identity
all replicon sequences identified in the 559 fully sequenced plasmids. For plasmid multilocus sequence typing (pMLST) analysis,
a database that is updated weekly was generated from www.pubmlst.org and integrated into a Web tool called pMLST. Both databases were evaluated using draft genomes from a collection of Salmonella enterica serovar Typhimurium isolates. PlasmidFinder identified a total of 103 replicons and between zero and five different plasmid
replicons within each of 49 S. Typhimurium draft genomes tested. The pMLST Web tool was able to subtype genomic sequencing data of plasmids, revealing
both known plasmid sequence types (STs) and new alleles and ST variants. In conclusion, testing of the two Web tools using
both fully assembled plasmid sequences and WGS-generated draft genomes showed them to be able to detect a broad variety of
plasmids that are often associated with antimicrobial resistance in clinically relevant bacterial pathogens.
Phylogenies are increasingly used in all fields of medical and biological research. Moreover, because of the next generation sequencing revolution, datasets used for conducting phylogenetic analyses grow at an unprecedented pace. RAxML (Randomized Axelerated Maximum Likelihood) is a popular program for phylogenetic analyses of large datasets under maximum likelihood. Since the last RAxML paper in 2006, it has been continuously maintained and extended to accommodate the increasingly growing input datasets and to serve the needs of the user community.
I present some of the most notable new features and extensions of RAxML, such as, a substantial extension of substitution models and supported data types, the introduction of SSE3, AVX, and AVX2 vector intrinsics, techniques for reducing the memory requirements of the code and a plethora of operations for conducting post-analyses on sets of trees. In addition, an up-to-date, 50 page user manual covering all new RAxML options is available.
The code is available under GNU GPL at https://github.com/stamatak/standard-RAxML.
Alexandros.Stamatakis@h-its.org.
Of all known Salmonella enterica serovars, S. Infantis is one of the most commonly isolated and has been recently emerging worldwide. To understand the recent emergence of S. Infantis in Israel we performed extensive comparative analyses between pre-emergent and the clonal emergent S. Infantis populations. We demonstrate the fixation of adaptive mutations in the DNA gyrase (gyrA) and nitroreductase (nfsA) genes, conferring resistance to quinolones and nitrofurans, respectively and the carriage of an emergent-specific plasmid, designated pESI. This self-transferred episome is a mosaic megaplasmid (∼280 kb), which increases bacterial tolerance to environmental mercury (mer operon), oxidative stress and provides further resistance to tetracycline, sulfamethoxazole and trimethoprim, most likely due to the presence of tetRA, sulI and dfrA genes, respectively. Moreover, pESI carries the yersiniabactin siderophore system and two novel chaperone-usher fimbriae. In-vitro studies established that pESI conjugation into a plasmidless S. Infantis strain results in superior biofilm formation, adhesion and invasion to avian and mammalian host cells. In-vivo mouse infections demonstrated higher pathogenicity and increased intestinal inflammation caused by a S. Infantis strain harboring pESI compared to the plasmidless parental strain. Our results indicate that the presence of pESI that was found only in the emergent population of S. Infantis in Israel contributes significantly to antimicrobials tolerance and pathogenicity of its carrier. It is highly likely that pESI plays a key role in the successful spread of the emergent clone that replaced the local S. Infantis community in short time of only 2-3 years.
Identification of antimicrobial resistance genes is important for understanding the underlying mechanisms and the epidemiology of antimicrobial resistance. As the costs of whole-genome sequencing (WGS) continue to decline, it becomes increasingly available in routine diagnostic laboratories and is anticipated to substitute traditional methods for resistance gene identification. Thus, the current challenge is to extract the relevant information from the large amount of generated data.
We developed a web-based method, ResFinder that uses BLAST for identification of acquired antimicrobial resistance genes in whole-genome data. As input, the method can use both pre-assembled, complete or partial genomes, and short sequence reads from four different sequencing platforms. The method was evaluated on 1862 GenBank files containing 1411 different resistance genes, as well as on 23 de-novo-sequenced isolates.
When testing the 1862 GenBank files, the method identified the resistance genes with an ID = 100% (100% identity) to the genes in ResFinder. Agreement between in silico predictions and phenotypic testing was found when the method was further tested on 23 isolates of five different bacterial species, with available phenotypes. Furthermore, ResFinder was evaluated on WGS chromosomes and plasmids of 30 isolates. Seven of these isolates were annotated to have antimicrobial resistance, and in all cases, annotations were compatible with the ResFinder results.
A web server providing a convenient way of identifying acquired antimicrobial resistance genes in completely sequenced isolates was created. ResFinder can be accessed at www.genomicepidemiology.org. ResFinder will continuously be updated as new resistance genes are identified.
Salmonella spp. remains one of the main pathogens causing diarrhea in humans worldwide. Lately, Salmonella Infantis has become endemic in several European, American, and Asian countries, presenting a multi-drug resistance profile and increased virulence. Various studies have attributed the high endemicity of Salmonella Infantis to pESI (plasmid to Emergent Salmonella Infantis). The ease of Salmonella to acquire pESI is of concern to health authorities and the food production chain. We searched for the presence of pESI in Salmonella genomes from the NCBI to understand the distribution of pESI worldwide and predict the main serovars and sequence types associated with the plasmid. We identified the pESI backbone, virulence, and resistance genes among Salmonella spp. isolated from 45 countries on five continents. We found the pESI-like structure in four different serovars: S. Muenchen, S. Schwarzengrund, S. Agona and S. Senftenberg. The pESI markers were also identified in 24 different sequence types. Most of the analyzed genomes were isolated from poultry, especially broiler and chicken. These results confirm the high dissemination of pESI-like megaplasmid among Salmonella Infantis worldwide and its ability to infect different serovars, as well as placing poultry production as the most favorable environment for pESI dissemination. Therefore, further studies are needed to prevent the spread of pESI to humans and the food chain.
Objectives
Horizontal acquisition of mobile genetic elements is a powerful evolutionary driving force that can profoundly affect pathogens epidemiology and their interactions with the environment and host. In the last decade, the role of the epidemic megaplasmid, pESI was demonstrated in the global emergence of multi-drug resistant (MDR) Salmonella enterica serovar Infantis strains, but it was unknown if this was a one-time phenomenon, or that pESI can drive the emergence of other pathogens.
Methods
Epidemiological, molecular, whole genome sequencing, de-novo assembly, bioinformatics and genetic approaches were used to analyze the emergence of a pESI-positive Salmonella enterica serovar Muenchen strain in Israel.
Results
Since 2018, we report the emergence and high prevalence of S. Muenchen in Israel, which consisted at 2020, 40% (1055/2671) of all clinical Salmonella isolates. We show that the emergence of S. Muenchen is dominated by a clonal MDR strain, report its complete assembled genome sequence, and demonstrate that in contrast to preemergent strains, it harbors the epidemic megaplasmid, pESI, which can be self-mobilized into E. coli and other Salmonella serovars. Additionally, we identified bioinformatically highly similar genomes of clinical isolates that were recently collected in South Africa, UK and USA.
Conclusions
This is a second documented case of a pathogen emergence associated with pESI acquisition. Considering the genetic cargo of pESI that enhances resistance, stress tolerance and virulence, and its ability to conjugate into prevalent Salmonella serovars, we provide further support that pESI facilities the emergence and spreading of new Salmonella strains.
Aims:
To characterize the genetic relatedness, phenotypic and genotypic antimicrobial resistance and plasmid content of 80 S. Infantis strains isolated from food, humans and veterinary sources from 2013 to 2018 in Brazil.
Methods and results:
PFGE and SNP analysis showed major clusters containing 50% and 38.8% of the strains studied, respectively. MLST assigned all strains to ST32. Disk-diffusion revealed that 90% of the strains presented resistant or intermediate resistant profiles and 38.8% displayed multidrug resistance. Resistance genes for aminoglycosides (aac(6')-Iaa; aadA12; aph(3'')-Ib; aph(6)-Id), β-lactams (blaTEM-1 ; blaCTX-M-8 ; blaCMY-2 ), trimethoprim (dfrA8), tetracycline (tet(A)), amphenicols (floR), sulfonamide (sul2), efflux pumps (mdsA; mdsB), chromosomal point mutations in gyrB, parC, acrB and pmrA were detected. Strains harbored IncI, IncF, IncX, IncQ, IncN and IncR plasmids.
Conclusions:
the presence of a prevalent S. Infantis subtype in Brazil and the high antimicrobial resistance rates reinforced the potential hazard of this serovar for the public health and food safety fields.
Significance and impact of the study:
this is the first study characterizing a large set of S. Infantis from Brazil by WGS, which provided a better local and global comprehension about the distribution and characteristics of this serovar of importance in the food, human and veterinary fields.
The objective of the present study was to characterize Salmonella enterica serovar Infantis isolated from chicken meat determining their clonal relationships with S. Infantis isolated from children with diarrhea. Fifteen meat-recovered S. Infantis were analyzed. Susceptibility levels to 14 antibacterial agents, the presence of ESBL and that of inducible plasmid-mediated AmpC (i-pAmpC) were determined by phenotypical methods. The presence of ESBL and pAmpC was confirmed by PCR, and detected ESBL-encoding genes were sequenced and their transferability tested by conjugation. The presence of gyrA mutations as well as Class 1 integrons was determined by PCR. Clonal relationships were established by REP-PCR and RAPD. In addition, 25 clinical isolates of S. Infantis were included in clonality studies. All meat-recovered S. Infantis were MDR, showing resistance to ampicillin, nitrofurans and quinolones, while none was resistant to azithromycin, ceftazidime or imipenem. ESBL (blaCTX-M-65) and i-pAmpC (blaDHA) were detected in 2 and 5 isolates respectively (in one case concomitantly), with blaCTX-M-65 being transferable through conjugation. In addition, 1 isolate presented a blaSHV gene. All isolates presented D87Y at GyrA, nalidixic acid active efflux pump and a Class 1 integron of ~1000 bp (aadA1). Clonal analysis showed that all isolates were related. Further they were identical to MDR blaCTX-M-65-producing S. Infantis isolates causing children diarrhea in Lima. The dissemination of MDR blaCTX-M-65-producing S. Infantis between marketed meat and children highlights a public health problem which needs be controlled at livestock level.
There has been an increase in the number of reports on Salmonella enterica subsp.
enterica serovar Infantis (S. Infantis) isolated from animals and humans. Recent studies using whole genome sequencing (WGS) have provided evidence on the likely contribution of a unique conjugative megaplasmid (pESI; ~280 kb) to the dissemination of this serovar
worldwide. In the present study, twenty-two unrelated Salmonella strains [S. Infantis (n=20) and Salmonella 6,7:r:- (n=2)] and their plasmids were investigated using next generation sequencing technologies (MiSeq and MinION) to unravel the significant expansion of this bacteria in Turkey. Multi-locus sequence typing, plasmid replicons, resistance gene contents as well as phylogenetic relations between strains were determined. According to the WGS
data, all S. Infantis possessed the relevant megaplasmid backbone genes and belonged to
sequence type 32 (ST32) with the exception of a single novel ST7091. Tetracycline and
trimethoprim/sulfamethoxazole resistance were found to be widespread in S. Infantis strains
and the resistant strains exclusively carried the tetA, sul1, sul2 and dfrA14 genes. One S.
Infantis isolate was also a carrier of the plasmid-mediated ampC via blaCMY-2, gene.
Moreover, full genomes of four S. Infantis isolates were reconstructed based on hybrid
assembly. All four strains contained large plasmids (240 – 290 kb) similar to previously
published megaplasmid (pESI) and accompanied by several small plasmids. The megaplasmid backbone contained a toxin-antitoxin system, two virulence cassettes and segments associated with heavy metals resistance, while variable regions possessed several antibiotic resistance genes flanked by mobile elements. This study indicated that pESI-like megaplasmid is widely disseminated within the tested S. Infantis strains of chicken meat, warranting further genomic studies on clinical strains from humans and animals to uncover the overall emergence and spread of this serovar.
Salmonella enterica subsp. enterica serovar Infantis (S. Infantis) isolates were found to have a multi-drug resistance profile (kanamycin, streptomycin, nalidixic acid, tetracycline, sulfonamide, and sometimes to ampicillin) and high prevalence (91%) in Turkish poultry in our previous studies. To investigate the mechanism behind multi-drug antimicrobial resistance (AMR) and high prevalence in Turkish poultry, 23 of the isolates were sequenced for comparative genomic analyses including: SNP-based comparison to S. Infantis from other countries, comparison of antimicrobial resistance genes (AMGs) with AMR phenotypes, and plasmid identification and annotation. Whole-genome SNP-based phylogenetic analysis found that all 23 Turkish S. Infantis isolates formed a distinct, well-supported clade, separate from 243 comparison S. Infantis genomes in GenomeTrakr identified as from the US and EU; the isolates most closely related to the cluster of these Turkish isolates were from Israel and Egypt. AMGs identified by bioinformatic analysis, without differentiating chromosomal or plasmid located genes, implied AMR phenotypes with 94% similarity overall to wet lab data, which was performed by phenotypic and conventional PCR methods. Most of the S. Infantis (21/23) isolates had identifiable plasmids, with 76% (16/21) larger than 100 kb and 48% (10/21) larger than 200 kb. A plasmid larger than 200 kb, with the incompatibility type of IncX1, similar to United States S. Infantis plasmid N55391 (99% query coverage and 99% identity overall), which itself is similar to Italian and Hungarian S. Infantis plasmids. Turkish S. Infantis plasmids had different beta-lactam resistance genes (blaTEM-70, blaTEM-148 and blaTEM-198) than the gene blaCTX-M-65 found in S. Infantis plasmids from other countries. This is the first observation of these three genes in S. Infantis isolates. The plasmids larger than 200 kb had two distinct regions of interest: Site 1 and Site 2. Site 1 (around 130 kb) had virulence- and bacteriocin- associated genes such as bacteriocin secretion system and type II toxin-antitoxin system genes (vagC, ccdA, ccdB, mchE, cvaB) and an aminoglycoside resistance gene (str). Site 2 (around 75-110 kb) had the antimicrobial resistance genes (aadA, sulI, tetA, tetR) and mercury (mer) resistance gene on tranposons Tn552 and Tn501. Presence of these AMR and virulence genes suggests they may have a role in the emergence of S. Infantis in poultry and support treating this serotype as a an important human health hazard.
Gastroenteritis in children is a serious condition in many parts of the world. Salmonella enterica is one of the causes of the disease. In this study, 280 fecal samples from children with diarrhea in four hospitals in Lima, Peru, were collected between September 2012 and March 2013. Salmonella was detected in 26 of the samples. Serotyping demonstrated that 25 of the isolates were S. enterica Infantis, and one isolate was S. enterica Typhimurium. Repetitive extragenic palindromic–polymerase chain reaction analysis suggests that all S. Infantis belong to the same clone. All but one of the S. Infantis isolates exhibited an extended-spectrum β-lactamase phenotype as they harbored bla CTX-M 65. Two strains also carried bla TEM-1. Nine of the isolates were resistant to azithromycin and two to ciprofloxacin. This study demonstrates that a multidrug-resistant S. Infantis clone carrying bla CTX-M 65 was circulating among children in Lima, Peru. The development of molecular epidemiology studies in Salmonella-causing diarrhea or other pathologies in Lima and in other areas will be useful to determine the permanence, geographical spread, and clinical implications of this clone.
Salmonella Infantis (SI) became endemic in Hungary where the PFGE cluster B, characterized by a large multiresistance (MDR) plasmid emerged among broilers leading to an increased occurrence in humans. We hypothesized that this plasmid (pSI54/04) assisted dissemination of SI. Indeed, Nal-Sul-Tet phenotypes carrying pSI54/04 occurred increasingly between 2011 and 2013 among SI isolates from broilers and humans. Characterization of pSI54/04 based on genome sequence data of the MDR strain SI54/04 indicated a size of ∼277 kb and a high sequence similarity with the megaplasmid pESI of SI predominant in Israel. Molecular characterization of 78 representative broiler and human isolates detected the prototype plasmid pSI54/04 and its variants of together with novel plasmid associations within the emerging cluster B. To test in vitro and in vivo pathogenicity of pSI54/04 we produced plasmidic transconjugant of the plasmid-free pre-emergent strain SI69/94. This parental strain and its transconjugant have been tested on chicken embryo fibroblasts (CEFs) and in orally infected day old chicks. The uptake of pSI54/04 did not increase the pathogenicity of the strain SI69/94 in these systems. Thus, dissemination of SI in poultry could be assisted by antimicrobial resistance rather than by virulence modules of the endemic plasmid pSI54/04 in Hungary.
Salmonella enterica subsp. enterica serovar Infantis has been reported to be carried asymptomatically in chickens and isolated from some human patients with diarrhea. The aim of this study was to investigate the phylogeny of S. Infantis strains isolated in Japan from chicken meat, chicken egg shells, environmental samples from a grading and packing center for chicken eggs (GP center), diarrhea patients, and asymptomatic carriers based on whole genome sequence data. The S. Infantis strains were in five clusters in a phylogenetic tree reconstructed by the maximum likelihood method. The five clusters were confirmed by neighbor-net and Bayesian cluster analyses. Two of the five clusters formed a group containing all of the strains isolated from chicken meat and some of the strains isolated from diarrhea patients and asymptomatic carriers. The median-joining network reconstructed in this study showed that strains in one of these two clusters diverged from one node with similar relatively short branches, suggesting clonal dissemination of these strains. The other three clusters formed a group containing all of the strains isolated from chicken egg shells and the GP center, and the remaining strains from diarrhea patients and asymptomatic carriers. Interestingly, strains isolated from patients did not cluster in only one group, indicating that none of the S. Infantis strains in this study had significantly higher human pathogenicity. The population genetic analyses in this study showed the separation of the five clusters into two groups was concordant with the sources where the strains in the clusters were isolated. These results suggested that evolutionary groups with higher hierarchy than the clusters identified in this study may be present, although such groups could not be determined by phylogenetic, neighbor-net, and Bayesian analyses in this study. Determination of higher level S. Infantis evolutionary groups should be investigated using other types of genetic markers.
To identify the mechanism(s) underlying cefotaxime resistance in 118 of 21 641 (0.55%) non-typhoidal Salmonella enterica isolates collected from humans throughout England and Wales from January 2010 to September 2012.
Non-duplicate isolates (n = 118) resistant to cefotaxime (MICs >1 mg/L) were screened by PCR for genes encoding CTX-M extended-spectrum β-lactamases (ESBLs) and associated ISEcp1-like elements, and for genes encoding acquired AmpC, SHV, TEM, VEB, PER and GES β-lactamases. Sequencing was used to identify specific alleles in selected isolates. Carbapenem resistance was sought by ertapenem disc screening.
Seventy-nine isolates (0.37% of all referred S. enterica) produced ESBLs, 37 isolates (0.17%) produced CMY-type AmpC enzymes, and 1 isolate had both enzyme types; the mechanism of cefotaxime resistance in 3 isolates could not be identified. Group 1 CTX-M genes were identified in 57 isolates belonging to 22 serotypes, with CTX-M-1 (n = 11), -15 (n = 9) and -55/57 (n = 8) the most prevalent alleles among the 29 (51%) investigated. CTX-M-2 (n = 5), -14 (n = 5), -8 (n = 1) and -65 (n = 1) were also identified. TEM-52 was identified in two isolates and SHV-12 in seven isolates. There was no evidence of carbapenem resistance. ESBL and AmpC genes were detected in both domestically acquired and travel-associated salmonellae. Eighty-nine isolates (75%) were multidrug resistant (resistant to at least three antimicrobial classes) and 42 (36%) had decreased susceptibility to ciprofloxacin (MICs 0.25-1 mg/L), with a further 13 (11%) isolates resistant (MICs >1 mg/L).
The prevalence of CTX-M and acquired AmpC genes in human non-typhoidal S. enterica from England and Wales is still low, but has increased from 0.03% in 2001-03 to 0.49% in 2010-12. Resistance to third-generation cephalosporins requires monitoring as it may reduce therapeutic options.
The National Food Surveillance System in Japan was formed in 1998 to monitor the contamination of retail foods with bacterial pathogens. Approximately 2000-3000 samples were tested annually, and the data from food categories that had more than 400 samples collected during 1998-2008 were analysed. With regard to meat, the frequency of positive samples for Salmonella in chicken for raw consumption and ground chicken was 12.7% and 33.5%, respectively. Moreover, Shiga toxin-producing Escherichia coli (STEC) O157 was found in ground meat, organ meat and processed meat, although at a low frequency (0.1%). The prevalence of Campylobacter jejuni/coli was 13.3% and 20.9% in chicken for raw consumption and ground chicken, respectively. In vegetables and fruit, Salmonella was detected in cucumber, lettuce, sprout and tomato samples at a frequency of around 0.1-0.2%. With regard to seafood, Salmonella was found in 0.5% of oysters for raw consumption. Seafood was not contaminated with STEC O157 or Shigella. Serotype Infantis was the most frequently detected serotype of Salmonella in seafood, followed by the serotypes Typhimurium, Schwarzengrund and Manhattan. In ground chicken, 72.2% of the strains were identified as the serotype Infantis. E. coli, as an indicator of food hygiene, was detected in all food categories. The results show the prevalence of the above-mentioned pathogens in the retail food supplied in Japan; further, they indicate that consumption of raw food carries the risk of contracting food-borne infections.
The European Union One Health 2019 Zoonoses Report.
- European Food Safety Authority
The European Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2016.
- European Food Safety Authority
Snippy: rapid haploid variant calling
- T Seemann