Search Results (10)

Background/Objectives: The acquisition of antimicrobial resistance by foodborne pathogens is a serious human health concern. In Japan, combinations of antimicrobial resistance genes in Salmonella from chicken meat were common among several serovars. Therefore, we hypothesized that different S. enterica serovars share a common antimicrobial resistance plasmid. Methods: Antimicrobial resistance transfer was tested in S. Infantis and S. Schwarzengrund, the major serovars used as donors. The plasmid structure was determined by subjecting S. Infantis Sal_238 and S. Schwarzengrund Sal_249 to short- and long-read sequencing. Results: The high homology between pSal_249Sch and pSal_238Inf suggests they have a common ancestor. Because the sequences of pSal_238Inf and pSal_249Sch were highly homologous to pESI (a plasmid for emerging S. Infantis), pSal_238Inf and pSal_249Sch were identified as pESI-like plasmids. S. Schwarzengrund is the third Salmonella serovar to expand its distribution related to pESI-like plasmid acquisition. Core-genome multilocus sequence-type analysis revealed that S. Schwarzengrund isolates with pESI-like plasmids from Japan (core-genome sequence-type [cgST] 167363 and cgST287831), the UK (cgST167363), and the USA (cgST167363, cgST196045, and cgST287831) were closely related; they are also suggested to share a common ancestor. The transfer of antimicrobial resistance was observed in combinations of both serovars. Specifically, the tentative plasmid sequence obtained via short-read sequencing, PCR, and conjugation experiments identified deletions of antimicrobial resistance genes (aadA, sul1, and tetA), class 1 integron, mercury resistance operon, and/or plasmid transfer region in the pESI-like plasmid. Conclusion: These data on the structural diversity of pESI-like plasmids suggest that some time has passed since S. Schwarzengrund acquired them. Full article

The global incidence of Salmonella enterica serovar Schwarzengrund has risen in recent years. This serotype has been isolated from poultry, retail meat, and other food products, leading to multiple outbreaks. Alongside the increase in infections, there are growing concerns about the increasing levels of antimicrobial resistance (AMR) among S. Schwarzengrund strains. This study aims to better understand the genetic factors possibly contributing to the rising prevalence of S. Schwarzengrund by analyzing the sequences of 2058 isolates from both human patients (N = 313) and food- and animal-associated sources, including chicken (N = 1145), turkey (N = 300), pork (N = 132), and other sources (N = 168). Data were obtained from GenBank and analyzed for AMR genes using AMRFinder. Additionally, putative virulence genes and plasmid transfer genes were assessed using the Virulence and AMR Plasmid Transfer Factor Database. AMR genes were found in 1269 (61.7%) of the isolates, with a total of 2478 AMR genes among the isolates, the most common being aph(3″)-Ib (N = 969, 47.1%), tet(A) (N = 190, 9.2%), and sul2 (N = 150, 7.3%), which are responsible for resistance to aminoglycoside, tetracycline, and sulfonamide, respectively. Additionally, 1060 (51.5%) isolates carried multiple plasmid transfer genes associated with IncFIB-FIC(FII) plasmids. Other plasmid types found in at least 1% of the strains included IncI1 (N = 101, 4.9%), IncHI2 (N = 62, 3.0%), or IncHI1 (N = 24, 1.2%). The virulence gene profiles of human isolates showed diversity but largely overlapped with those from different food sources. Notably, the aerobactin iron acquisition genes, associated with Salmonella’s virulence and colonization, were highly prevalent among chicken isolates (N = 1019, 89.0%) but less frequent in isolates from other sources (N = 65, 7.2%). IncFIB-FIC(FII) plasmids, commonly harboring the aerobactin operon, were highly prevalent among chicken-related isolates and present in about 10% of human isolates. The diverse plasmid, AMR, and virulence gene profiles in human-associated isolates suggest that multiple factors may contribute to the increased virulence in S. Schwarzengrund. Full article

Salmonella enterica is a globally important zoonotic microorganism that affects pigs and can enter the farm through various routes. This study aimed to determine the prevalence of S. enterica in water sources and pigs at Colombian pig farms, and to characterize the antimicrobial resistance of the isolates phenotypically and genotypically. Samples were collected from 103 farms including source water (n = 104), storage tank water (n = 103), drinking water (n = 103), and individual rectal swab samples (n = 1025). The presence of Salmonella was detected/identified using MDS-3M™ agar culture medium. Isolates were serotyped, and their antibiotic susceptibility was determined by minimum inhibitory concentration (MIC). Whole genome sequencing (WGS) was performed using Illumina NovaSeq, and bioinformatics analysis focused on serovar confirmation, MLST determination, and resistance gene detection. The overall between-farm prevalence of Salmonella enterica including all types of samples was 52.4% (54/103), with 6.4% of rectal swab samples and 21.3% of water samples found to be positive. Thirty serovars were identified using WGS, with the most common being S. Typhimurium var. monophasic (1,4,[5],12:i:-) (41.2%), S. Schwarzengrund (4.2%), and S. Saintpaul (4.2%). Salmonella Typhimurium and its monophasic variant were more commonly found in rectal swabs than the remaining serotypes (relative risk = 2.9, p < 0.0001), which were commonly found in the water samples (relative risk = 5.2, p < 0.0001). High levels of phenotypic resistance were observed, particularly to amikacin (99.2%), tetracycline (59.7%), chloramphenicol (55.5%), and ampicillin (42%). All isolates carried genes conferring resistance to aminoglycosides (aac(6′)-Iaa), quinolones (qnrB19), and tetracyclines (tetA). In conclusion, S. enterica is prevalent in Colombian pig farms including the water supply, with the S. Typhimurium monophasic variant being predominant, and antimicrobial resistance is widespread. Full article

Microbial resistance to antibiotics poses a significant threat to both human and animal health, necessitating international efforts to mitigate this issue. This study aimed to assess the resistance profiles of Salmonella sp. isolates and identify the presence of intl1, sul1, and blaTEM resistance genes within antigenically characterized isolates, including Agona, Livingstone, Cerro, Schwarzengrund, Salmonella enterica subsp. enterica serotype O:4.5, Anatum, Enteritidis, Johannesburg, Corvallis, and Senftenberg. These isolates underwent susceptibility testing against 14 antibiotics. The highest resistance percentages were noted for sulfamethoxazole (91%), sulfonamides (51%), and ceftiofur (28.9%), while no resistance was observed for ciprofloxacin. Salmonella Johannesburg and Salmonella Corvallis showed resistance to one antibiotic, whereas other serovars were resistant to at least two. Salmonella Schwarzengrund exhibited resistance to 13 antibiotics. The intl1 gene was detected in six out of the ten serovars, and the sul1 gene in three, always co-occurring with intl1. The blaTEM gene was not identified. Our findings highlight the risk posed by the detected multiple resistances and genes to animal, human, and environmental health. The multidrug resistance, especially to third-generation cephalosporins and fluoroquinolones, highlights the need for stringent monitoring of Salmonella in laying hens. The potential of the environment, humans, eggs, and their products to act as vectors for antibiotic resistance represents a significant concern for One Health. Full article

This study’s goal was to determine the prevalence, temporal trends, seasonal patterns, and temporal clustering of Salmonella enterica isolated from environmental samples from Ontario’s poultry breeding flocks between 2009 and 2018. Clusters of common serovars and those of human health concern were identified using a scan statistic. The period prevalence of S. enterica was 25.3% in broiler breeders, 6.4% in layer breeders, and 28.6% in turkey breeders. An overall decreasing trend in S. enterica prevalence was identified in broiler breeders (from 27.8% in 2009 to 22.1% in 2018) and layer breeders (from 15.4% to 4.9%), while an increasing trend was identified in turkey breeders (from 12.0% to 24.5%). The most common serovars varied by commodity. Among broiler breeders, S. enterica serovars Kentucky (42.4% of 682 submissions), Heidelberg (19.2%), and Typhimurium (5.4%) were the most common. Salmonella enterica serovars Thompson (20.0% of 195 submissions) and Infantis (16.4%) were most common among layer breeders, and S. enterica serovars Schwarzengrund (23.6% of 1368 submissions), Senftenberg (12.9%), and Heidelberg and Uganda (9.6% each) were most common among turkey breeders. Salmonella enterica ser. Enteritidis prevalence was highest in submissions from broiler breeders (3.7% of 682 broiler breeder submissions). Temporal clusters of S. enterica serovars were identified for all poultry commodities. Seasonal effects varied by commodity, with most peaks occurring in the fall. Our study provides information on the prevalence and temporality of S. enterica serovars within Ontario’s poultry breeder flocks that might guide prevention and control programs at the breeder level. Full article

It is well established that plasmids carrying multiple antimicrobial resistance (AMR) genes can be easily transferred among bacterial isolates by horizontal gene transfer. Previous studies have shown that a combination of short- and long-read approaches is effective in reconstructing accurate plasmids. However, high-quality Illumina short reads mapped onto the long reads in the context of an AMR hybrid monitoring strategy have not yet been explored. Hence, this study aimed to improve the reconstruction of plasmids, including the localization of AMR genes, using the above-described parameters on whole-genome sequencing (WGS) results. To the best of our knowledge, this study is the first to use S1 nuclease pulsed-field gel electrophoresis (S1-PFGE) to confirm the number and sizes of plasmids detected by in silico-based predictions in Salmonella strains. Our results showed that de novo assembly did not detect the number of bacterial plasmids more accurately than reference-based assembly did. As this new hybrid mapping strategy surpassed de novo assembly in bacterial reconstruction, it was further used to identify the presence and genomic location of AMR genes among three Salmonella enterica serovar Schwarzengrund isolates. The AMR genes identified in the bacterial chromosome among the three Salmonella enterica serovar Schwarzengrund isolates included: AAC(3)-IV, AAC(6′)-Iy, aadA2, APH(4)-Ia, cmlA1, golS, mdsA, mdsB, mdsC, mdtK, qacH, sdiA, sul2, sul3, and TEM-1 genes. Moreover, the presence of TEM-1, AAC(3)-IV, aadA2, APH(4)-Ia, cmlA1, dfrA12, floR, sul1, sul3, and tet(A) genes found within three IncFIB plasmids and one IncX1 plasmid highlight their possible transmission into the environment, which is a public health risk. In conclusion, the generated data using this new hybrid mapping strategy will contribute to the improvement of AMR monitoring and support the risk assessment of AMR dissemination. Full article

Our previous study revealed that Salmonella enterica serovar Schwarzengrund-contaminated areas of broiler chickens have expanded from West Japan to East Japan. The present study investigated the antimicrobial resistance and molecular characteristics of 124 S. Schwarzengrund isolates obtained from chicken meat produced in East and West Japan from 2008 to 2019. Comparing the isolates obtained in 2008 and 2015–2019, an increase in the proportion of those resistant to kanamycin [51.4–89.7% (p < 0.001)] was observed. In contrast, the proportion of isolates resistant to both streptomycin and tetracycline and those that harbored a 1.0-kb class 1 integron, aadA1, and tetA, significantly decreased from 100% in 2008 to 47.1% in 2015–2019 (p < 0.001). A 1.0-kb class 1 integron containing aadA1, harbored by 78 isolates, was different from that reported in globally distributed S. Schwarzengrund strains (1.9 kb, containing the dfrA12-aadA2 gene cassette). Twenty-five isolates from different product districts and years of isolation were typed as sequence type (ST) 241 with multilocus sequence typing. Our results suggest that S. Schwarzengrund, which contaminates chicken meat in Japan, shares a common ancestor regardless of the product district from 2008 to recent years. Moreover, S. Schwarzengrund ST241 may have spread from western to eastern Japan. Full article

Over the last decade, Salmonella enterica serovar Schwarzengrund has become more prevalent in Asia, Europe, and the US with the simultaneous emergence of multidrug-resistant isolates. As these pathogens are responsible for many sporadic illnesses and chronic complications, as well as outbreaks over many countries, improved surveillance is urgently needed. For 20 years, pulsed-field gel electrophoresis (PFGE) has been the gold standard for determining bacterial relatedness by targeting genome-wide restriction enzyme polymorphisms. Despite its utility, recent studies have reported that PFGE results correlate poorly with that of closely related outbreak strains and clonally dominant endemic strains. Due to these concerns, alternative amplification-based molecular methods for bacterial strain typing have been developed, including clustered regular interspaced short palindromic repeats (CRISPR) and multilocus sequence typing (MLST). Furthermore, as the cost of sequencing continues to decrease, whole genome sequencing (WGS) is poised to replace other molecular strain typing methods. In this study, we assessed the discriminatory power of PFGE, CRISPR, MLST, and WGS methods to differentiate between 23 epidemiologically unrelated S. enterica serovar Schwarzengrund isolates collected over an 18-year period from distinct locations in Taiwan. The discriminatory index (DI) of each method for different isolates was calculated, resulting in values between 0 (not discriminatory) and 1 (highly discriminatory). Our results showed that WGS has the greatest resolution (DI = 0.982) compared to PFGE (DI = 0.938), CRISPR (DI = 0.906), and MLST (DI = 0.463) methods. In conclusion, the WGS typing approach was shown to be the most sensitive for S. enterica serovar Schwarzengrund fingerprinting. Full article

Salmonella enterica serovar Schwarzengrund is one of the most frequently isolated Salmonella serotypes responsible for human and poultry infections in Taiwan, and it has raised public health concerns. To better facilitate the understanding of transmission patterns and the dynamics of epidemics, sharing molecular data on pathogen profiles is urgently needed. The objectives of the current study were to determine and establish baseline data of S. enterica serovar Schwarzengrund isolates from 23 epidemiologically unrelated sources from year 2000 to 2018 and examine their phenotypic and genotypic characteristics. Genomic DNA of the Salmonella isolates was extracted and subjected to whole-genome sequencing using an Illumina platform. Results showed that all selected isolates exhibited multidrug resistance, and six of those were resistant to ciprofloxacin phenotypically. Genotypically, these isolates carried genes resistant to aminoglycoside (100%), phenicol (91.3%), β-lactams (69.5%), folate pathway antagonist (100%), tetracycline (82.6%), and fluoroquinolone (4.3%). Moreover, these isolates harbor integrons with five different gene cassettes identified for the first time, which are associated with resistance to trimethoprim, streptomycin, tetracycline, sulfonamide, chloramphenicol, and gentamicin. Furthermore, prevalence of IncFIB plasmid was found among studied isolates, which may increase its ability to colonize the chicken cecum and cause extra-intestinal disease. Salmonella pathogenicity islands SPI-1 to SPI-5, SPI-13, and SPI-14, as well as C63PI locus, were also detected in all isolates. This study demonstrated that a considerable high antimicrobial resistance with high virulence levels of Salmonella were found from animal sources. Sharing data on these pathogen profiles can not only help increase the reproducibility and accessibility of genomic analysis but can also support surveillance and epidemiological investigations for salmonellosis in the region. Full article

Salmonella enterica is a major cause of gastroenteritis usually caused by animal-based contaminated foods. Since the current passive surveillance is not sufficient to detect all infections and infection sources, we determined the prevalence of Salmonella isolated from sewage influent of wastewater treatment plants (WWTPs) and compared the characteristics of human and food isolates to identify the infection sources. Sewage influent samples were collected monthly from two WWTPs located in the Yamanashi Prefecture, Japan, for three years. Serotypes, antimicrobial resistances, isolation periods, isolated areas, and pulsed-field gel electrophoresis patterns of six isolates belonging to five serotypes were consistent with those of the isolates from patients. Real-time PCR for Salmonella indicated that sewage influents reflect cases of patients infected with Salmonella, including unreported cases. Serovars Schwarzengrund and Anatum were predominant in sewage, but not in humans, and their characteristics were closely related or identical to those isolated from poultry heart and liver, respectively. These results suggest that sewage influent contains Salmonella isolates from humans and that some originated from unreported human cases infected by poultry-associated products. Therefore, it is necessary to take countermeasures against Salmonella infection based on the unreported cases, which would be disclosed by analysis of sewage influent. Full article