Salmonella enterica serotypes from human and nonhuman sources in Sao Paulo State, Brazil, 2004-2020.

Salmonellosis ranks among the most frequently reported zoonosis worldwide and is often associated with foodborne outbreaks. Since the 1950s, the distribution of Salmonella serotypes in Sao Paulo State, Brazil, has been documented and periodically reported. In this study, we updated the data on the distribution of Salmonella serotypes received in our reference laboratory, isolated from human infections and nonhuman sources, from 2004 to 2020. In that period, a total of 9,014 Salmonella isolates were analyzed, of which 3,553 (39.4%) were recovered from human samples, mainly of stool (65%) and blood (25.6%), and 5,461 (60.6%) were isolated from nonhuman origins, such as animals (47.2%), food (27.7%) and animal environments (18.6%). In human isolates, a total of 104 serotypes were identified and the most frequent ones were Enteritidis, Typhimurium, S . I. 4,[5],12:i:-, Dublin and Typhi. A consistent reduction of the Enteritidis proportion was observed over the years. Among the 156 serotypes identified in isolates with nonhuman origins, Enteritidis, Mbandaka, Typhimurium, Agona and Anatum were ranked as the top five Salmonella serotypes; in more recent years, S . Heidelberg has increased in frequency. Although with different proportions, the top 10 prevalent serotypes were identified in both human and nonhuman origins, underscoring the role of animals, food products and environment as reservoirs of Salmonella with potential to cause human salmonellosis.

Salmonella enterica serotypes from human and nonhuman sources in Sao Paulo State, Brazil, 2004-2020

ABSTRACT Salmonellosis ranks among the most frequently reported zoonosis worldwide and is often associated with foodborne outbreaks. Since the 1950s, the distribution of Salmonella serotypes in Sao Paulo State, Brazil, has been documented and periodically reported. In this study, we updated the data on the distribution of Salmonella serotypes received in our reference laboratory, isolated from human infections and nonhuman sources, from 2004 to 2020. In that period, a total of 9,014 Salmonella isolates were analyzed, of which 3,553 (39.4%) were recovered from human samples, mainly of stool (65%) and blood (25.6%), and 5,461 (60.6%) were isolated from nonhuman origins, such as animals (47.2%), food (27.7%) and animal environments (18.6%). In human isolates, a total of 104 serotypes were identified and the most frequent ones were Enteritidis, Typhimurium, S . I. 4,[5],12:i:-, Dublin and Typhi. A consistent reduction of the Enteritidis proportion was observed over the years. Among the 156 serotypes identified in isolates with nonhuman origins, Enteritidis, Mbandaka, Typhimurium, Agona and Anatum were ranked as the top five Salmonella serotypes; in more recent years, S . Heidelberg has increased in frequency. Although with different proportions, the top 10 prevalent serotypes were identified in both human and nonhuman origins, underscoring the role of animals, food products and environment as reservoirs of Salmonella with potential to cause human salmonellosis.

Subtyping of plasmid-mediated quinolone resistance among Salmonella serotypes by whole genome sequencing.

Most known plasmids are identified by conferring virulence or antimicrobial resistance phenotypes and such characteristics aid in the success of the dispersion of different plasmid types between bacteria from different sources. This study aimed to perform the subtyping of the plasmid-mediated quinolone resistance, detected in Salmonella spp. A total of 34 Salmonella strains non-susceptible to ciprofloxacin were evaluated. Strains were selected based on the presence of PMQR determined by Polymerase Chain Reaction and further submitted to Next Generation Sequencing. Most of the strains presented the qnrB19 in small ColE-like plasmids and qnrB2 gene associated with IncN/ST5 plasmids also detected. Our results indicated the co-occurrence of PMQR and ESBLs in plasmids that are a lineage of epidemic plasmids circulating in Salmonella in which additional resistances were detected, highlighting the potential threat of resistance Salmonella to public health, particularly in infections in which antimicrobial therapy is needed.

Molecular analysis of clonally related Salmonella Typhi recovered from epidemiologically unrelated cases of typhoid fever, Brazil.

BACKGROUND: The primary method of molecular subtyping for the identification and investigation of outbreaks has been pulsed-field gel electrophoresis (PFGE). In some cases, this technique has not been able to show discrimination between the unrelated strains that can be achieved by whole genome sequencing (WGS). METHODS: The aim of this study was to determine the strengths and drawbacks of WGS using different analytic approaches compared to traditional typing method, PFGE, for retrospectively typing clusters cases of 28 S. Typhi. RESULTS: We evaluated three analytical approaches on the WGS data set (Nucleotide Difference (ND), (SNPs) and Whole genome multi locus sequence typing (wgMLST) that identically classified the clusters-related strains into two clusters, cluster A (with strains from 2017), and Cluster B (with strains from 2007). CONCLUSIONS: In this study WGS based typing, was able to compete with PFGE for differentiation of the clusters of S. Typhi strains.

Emergence of CMY-2-Producing Salmonella Heidelberg Associated with IncI1 Plasmids Isolated from Poultry in Brazil.

In this study we report the characterization of plasmid-mediated CMY-2-producing Salmonella Heidelberg recovered from food, poultry, and poultry environment in Brazil, between 2014 and 2016. The blaCMY-2 resistance gene was allocated in large (90-148 kb) IncI1 type transferable plasmids. Salmonella Heidelberg isolates were genetically related, indicating the dissemination of closely related isolates among food, poultry, and its environment. This is the first report of IncI1 replicon-types of plasmids encoding the blaCMY-2 resistance gene in Salmonella Heidelberg isolates in Brazil, the world's biggest exporter of chickens.

Genotypic Resistance to Quinolone and Tetracycline in Salmonella Dublin Strains Isolated from Humans and Animals in Brazil.

Resistance of Salmonella Dublin strains to quinolones and tetracycline has been increasing worldwide. Studies regarding the genotypic resistance traits of strains of this serovar isolated in Brazil are scarce. This study aims to examine the genetic characteristics of Salmonella Dublin strains isolated in Brazil, which are associated with resistance to quinolone and tetracycline. The minimum inhibitory concentrations (MICs) of nalidixic acid, ciprofloxacin, and tetracycline of the 10 strains sensitive and 21 strains resistant to quinolone and tetracycline were determined using Etest.® The mutation profiles of the gyrA, gyrB, parC, and parE genes were accessed by sequencing, while the presence of plasmid-mediated quinolone resistance and tet genes was analyzed by PCR. Quinolone-resistant strains presented the amino acid substitutions Ser96→Tyr, Ser96→Phe, Asp107→Asn, or Asp108→Gly on the gyrA gene, and the Ser224→Phe and Glu231→Asp mutations on the gyrB gene. The qnrA, tet(A), and tet(B) genes were detected in 5, 13, and 6 strains, respectively. Analysis of the MIC values revealed that 1 and 3 strains presented intermediate and resistant MIC profiles to nalidixic acid, respectively; 6 strains presented intermediate MIC profile to ciprofloxacin; and 13 strains presented resistant MIC profile to tetracycline. In the Salmonella Dublin strains studied, quinolone resistance was mainly related to mutation points that led to target alteration in the gyrA and gyrB genes, while tetracycline resistance was associated with the presence of tet(A) and/or tet(B) genes, with the highest resistance levels detected in strains bearing the tet(B) gene. The presence of the aforementioned genotypic resistance traits in Salmonella Dublin strains isolated over 33 years in Brazil indicates that ciprofloxacin or tetracycline therapy against such strains may fail.

Subtyping of plasmid-mediated quinolone resistance among Salmonella serotypes by whole genome sequencing

Most known plasmids are identified by conferring virulence or antimicrobial resistance phenotypes and such characteristics aid in the success of the dispersion of different plasmid types between bacteria from different sources. This study aimed to perform the subtyping of the plasmid-mediated quinolone resistance, detected in Salmonella spp. A total of 34 Salmonella strains non-susceptible to ciprofloxacin were evaluated. Strains were selected based on the presence of PMQR determined by Polymerase Chain Reaction and further submitted to Next Generation Sequencing. Most of the strains presented the qnrB19 in small ColE-like plasmids and qnrB2 gene associated with IncN/ST5 plasmids also detected. Our results indicated the co-occurrence of PMQR and ESBLs in plasmids that are a lineage of epidemic plasmids circulating in Salmonella in which additional resistances were detected, highlighting the potential threat of resistance Salmonella to public health, particularly in infections in which antimicrobial therapy is needed.

Subtyping of plasmid-mediated quinolone resistance among Salmonella serotypes by whole genome sequencing

Most known plasmids are identified by conferring virulence or antimicrobial resistance phenotypes and such characteristics aid in the success of the dispersion of different plasmid types between bacteria from different sources. This study aimed to perform the subtyping of the plasmid-mediated quinolone resistance, detected in Salmonella spp. A total of 34 Salmonella strains non-susceptible to ciprofloxacin were evaluated. Strains were selected based on the presence of PMQR determined by Polymerase Chain Reaction and further submitted to Next Generation Sequencing. Most of the strains presented the qnrB19 in small ColE-like plasmids and qnrB2 gene associated with IncN/ST5 plasmids also detected. Our results indicated the co-occurrence of PMQR and ESBLs in plasmids that are a lineage of epidemic plasmids circulating in Salmonella in which additional resistances were detected, highlighting the potential threat of resistance Salmonella to public health, particularly in infections in which antimicrobial therapy is needed.

Draft Genome Sequences of 112 Salmonella enterica Serovar Dublin Strains Isolated from Humans and Animals in Brazil.

Salmonella enterica serovar Dublin is a strongly adapted serovar that causes enteritis and/or systemic disease in cattle and results in high rates of mortality. Here, we report the draft genome sequences of 112 S. Dublin strains isolated from humans and animals in Brazil. These draft genome sequences will help enhance our understanding of this serovar in Brazil.

Prevalence of Extended-Spectrum ß-Lactamases CTX-M-8 and CTX-M-2-Producing Salmonella Serotypes from Clinical and Nonhuman Isolates in Brazil.

We characterized extended-spectrum ß-lactamases (ESBL) enzymes among Salmonella strains isolated in Brazil from 2009 to 2014. Salmonella recovered from both clinical and nonhuman (food, poultry, and environment) sources were subjected to antimicrobial susceptibility testing. ß-lactamases genes were detected by polymerase chain reaction/sequencing; plasmid profiles and transferability were assessed by S1-pulsed field gel electrophoresis (PFGE). Genetic diversity was evaluated by XbaI-PFGE. Out of 630 Salmonella strains screened, 46 displayed ESBL phenotype, distributed across 11 different serotypes. blaCTX-M-8 and blaCTX-M-2 genes were detected at frequencies of 47% and 41%, respectively. blaSHV-5 and blaSHV-2 were also detected but in lower frequencies (4%, 2%). blaTEM-1 gene was detected in 22% of the strains. Most of the ESBL genes were transferable by conjugation, and the respective blaESBL gene was detected in the recipient strain, indicating the location of ESBL determinants on transferable plasmids. XbaI-PFGE revealed genomic diversity of Salmonella Typhimurium bearing blaCTX-M-2, blaCTX-M-8, blaTEM-1, and blaSHV-2 genes. Salmonella Muenchen (harboring blaCTX-M-2) and Salmonella Corvallis (blaCTX-M-8 and blaSHV-5) showed clonal relatedness within respective serotypes. Our findings underscore the occurrence of diverse ESBL genes in several Salmonella serotypes, reinforcing the need for continuous surveillance of resistance genes circulating in human and nonhuman sources.

Presence of plasmid-mediated quinolone resistance determinants and mutations in gyrase and topoisomerase in Salmonella enterica isolates with resistance and reduced susceptibility to ciprofloxacin.

In recent decades, the emergence and spread of resistance to nalidixic acid are usually associated with reduced susceptibility to ciprofloxacin among Salmonella serotypes. The aims of this study were to investigate the mechanisms associated with resistance to fluoroquinolone and the clonal relatedness of Salmonella strains isolated from human and nonhuman origins, in a 5-year period in São Paulo, Brazil. Antimicrobial susceptibility testing for Salmonella isolates was performed. PCR and DNA sequencing were accomplished to identify mutations in the quinolone resistance-determining regions of the topoisomerase genes and to determine the fluoroquinolone determinants. The strains presented MIC to ciprofloxacin ranging from 0.125 to 8.0 mg/L (all nonsusceptible). From these, 16 strains (17.5%) were resistant to ciprofloxacin (MIC ≥1 mg/L) and belonging to serotypes Typhimurium, I. 4,5,12:i:-, Enteritidis, and Heidelberg. Amplification and DNA sequencing of topoisomerases genes identified multiple amino acid substitutions in GyrA and ParC. No mutations were identified in GyrB, and 1 amino acid substitution was identified in ParE. Among the 16 Salmonella strains resistant to ciprofloxacin, 8 S. I. 4,5,12:i:- presenting mutations in gyrA and parE genes were grouped into the same pulsotype. Plasmid-mediated quinolone resistance (PMQR) determinants: qnrB, aac(6')-lb-cr, and oqxA/B were detected among 13 strains. To the best of our knowledge, this is the first work to report Salmonella isolates resistant to ciprofloxacin in Brazil. Indeed, this is the first detection of PMQR determinants in Salmonella strains from Sao Paulo State. These findings alert for the potential spread of quinolone resistance of Salmonella strains, particularly in S. I. 4,5,12:i:-, a prevalent serotype implicated in human disease and foodborne outbreaks.

Salmonella Alachua: causative agent of a foodborne disease outbreak

Objectives: The aim of this study is to report the occurrence of the first outbreak of food poisoning caused by Salmonella Alachua in Brazil, as well as the antimicrobial susceptibility and the genetic relatedness of Salmonella Alachua strains isolated from clinical and food samples. Material and methods: To elucidate the outbreak, an epidemiological investigation was carried out, and two samples of common food were tested - mayonnaise salad and galinhada (a traditional Brazilian dish of chicken and rice) - according to the Compendium of methods for the microbiological examination of foods. Five stool samples were tested employing classic methods for the isolation and identification of enterobacteria. Strains of Salmonella were characterized for antibiotic susceptibility according to the Clinical and Laboratory Stan- dards Institute guidelines (2013), and submitted to pulsed-field gel electrophoresis analysis, performed according to the Centers for Disease Control and Prevention PulseNet protocol. Results: A total of 94 people were interviewed after ingesting the food, 66 of whom had become ill. A 60-year old female patient who was hospitalized in a serious condition, developed septic shock and died two days after consuming the food. The presence of Salmonella Alachua was confirmed in all the analyzed stool samples, and in the two types of food. The five strains showed higher than minimum inhibitory concentration values of nalidixic acid (≥256 µg/mL) and reduced ciprofloxacin susceptibility (minimum inhibitory concentration = 0.5 µg/mL). The pulsed-field gel electrophoresis analysis revealed indistinguishable patterns in Salmonella Alachua strains isolated from clinical and food samples. Conclusion: The data presented herein confirm the foodborne disease outbreak. They also allowed for the identification of the source of infection, and suggest that products from poultry are potential reservoirs for this serotype, reinforcing the ...

Salmonella Alachua: causative agent of a foodborne disease outbreak.

OBJECTIVES: The aim of this study is to report the occurrence of the first outbreak of food poisoning caused by Salmonella Alachua in Brazil, as well as the antimicrobial susceptibility and the genetic relatedness of Salmonella Alachua strains isolated from clinical and food samples. MATERIAL AND METHODS: To elucidate the outbreak, an epidemiological investigation was carried out, and two samples of common food were tested--mayonnaise salad and galinhada (a traditional Brazilian dish of chicken and rice)--according to the Compendium of methods for the microbiological examination of foods. Five stool samples were tested employing classic methods for the isolation and identification of enterobacteria. Strains of Salmonella were characterized for antibiotic susceptibility according to the Clinical and Laboratory Standards Institute guidelines (2013), and submitted to pulsed-field gel electrophoresis analysis, performed according to the Centers for Disease Control and Prevention PulseNet protocol. RESULTS: A total of 94 people were interviewed after ingesting the food, 66 of whom had become ill. A 60-year old female patient who was hospitalized in a serious condition, developed septic shock and died two days after consuming the food. The presence of Salmonella Alachua was confirmed in all the analyzed stool samples, and in the two types of food. The five strains showed higher than minimum inhibitory concentration values of nalidixic acid (≥256 µg/mL) and reduced ciprofloxacin susceptibility (minimum inhibitory concentration=0.5 µg/mL). The pulsed-field gel electrophoresis analysis revealed indistinguishable patterns in Salmonella Alachua strains isolated from clinical and food samples. CONCLUSION: The data presented herein confirm the foodborne disease outbreak. They also allowed for the identification of the source of infection, and suggest that products from poultry are potential reservoirs for this serotype, reinforcing the importance of warning consumers about the danger of possible contamination.

Antimicrobial susceptibility, phage types, and pulse types of Salmonella Typhimurium, in São Paulo, Brazil.

A total of 283 Salmonella Typhimurium strains isolated from cases of human infections and non human sources, were examined for antimicrobial susceptibility and the incidence of resistance was 38% and multiple resistance (to three or more antimicrobials) was 15%. All 43 multidrug-resistant strains (MDR) and 13 susceptible ones were characterized by phage typing and pulsed- field gel electrophoresis (PFGE). The strains encompassed 14 definitive phage types (DT), three were untypable (UT), and 18 atypicals or reaction does not conform (RDNC), which belonged to 21 PFGE patterns, A1-A21. The predominant phage types were DT49, DT193, and RDNC and two strains belonging to DT 104 and 104b were also identified. The most common PFGE patterns were A1 and A8. Analysis by PFGE and phage typing demonstrated that the most of the MDR were multiclonal and association among multiresistance, phage typing, and PFGE patterns was not so significant.

Antimicrobial susceptibility, phage types, and pulsetypes of Salmonella Typhimurium, in São Paulo, Brazil

A total of 283 Salmonella Typhimurium strains isolated from cases of human infections and non human sources, were examined for antimicrobial susceptibilityand the incidence of resistance was 38 percent and multiple resistance (to three or more antimicrobials) was 15 percent. All 43 multidrug-resistant strains (MDR) and 13 susceptible ones were characterized by phage typing and pulsed- field gel electrophoresis (PFGE). The strains encompassed 14 definitive phage types (DT), three were untypable (UT), and 18 atypicals or reaction does not conform (RDNC), which belonged to 21 PFGE patterns, A1-A21. The predominant phage types were DT49, DT193, and RDNC and two strains belonging to DT 104 and 104b were also identified. The most commum PFGE patterns were A1 and A8. Analysis by PFGE and phage typing demonstrated that the most of the MDR were multiclonal and association among multiresistance, phage typing, and PFGE patterns was not so significant.

Ribotyping of Salmonella Enteritidis strains reveals the spread of a single genotype in the Brazilian city of Ribeirão Preto

INTRODUÇAO: Na identificação de cepas de Salmonella, os métodos de sorotipagem e ribotipagem na detecção de marcadores epidemiológicos são os mais utilizados nos laboratórios de referência mundiais. Esses métodos moleculares são imprescindíveis na vigilância epidemiológica e permitem a detecção da fonte da infecção. OBJETIVOS: O presente trabalho objetivou caracterizar as cepas de Salmonella Enteritidis pela ribotipagem. MATERIAL E MÉTODOS: Trinta e oito cepas de S. Enteritidis foram isoladas de pacientes atendidos no Hospital das Clínicas da Universidade de São Paulo, Ribeirão Preto, SP, entre os anos de 1996 e 1998. As cepas foram isoladas de fezes (31 amostras), sangue (quatro amostras) e outros fluidos (três amostras). As cepas de Salmonella foram isoladas utilizando-se métodos bacteriológicos de rotina, sorotipadas e ribotipadas. RESULTADOS: As 38 cepas de S. Enteritidis apresentaram na ribotipagem a separação das cepas em dois ribotipos: A (94,7 por cento das amostras) e B (5,3 por cento das amostras). DISCUSSAO E CONCLUSAO: Esses dados sugerem que grande parte dos pacientes (94,7 por cento) foi infectada pela mesma cepa. Essa cepa pode ser endêmica na comunidade de Ribeirão Preto ou os pacientes foram expostos a uma fonte comum de infecção.

Toxinfecção alimentar por Salmonella em evento científico, São Paulo/SP, setembro de 2004 / Food toxinfection by Salmonella in scientific event at São Paulo/SP, september 2004

Identificar e investigar precocemente surtos causados por alimentos é um dos componentes essenciais da vigilância epidemiológica das doenças transmitidas por alimentos. Doenças veiculadas por alimentos representam, atualmente, na maior parte dos países, a principal causa de surtos de diarréia. A globalização da economia, as alterações nos hábitos alimentares, o crescente consumo de alimentos preparados fora de casa, dentre outros aspectos, alteraram o perfil epidemiológico dessas doenças, expondo a população a vários tipos de contaminantes. Por sua vez, o desemprego tem sido um fator importante para o crescimento do setor de prestação de serviços alimentícios, do comércio ambulante de alimentos e mesmo de bufês, principalmente nos países em desenvolvimento; e não apenas permanecem como clandestinos perante a legislação sanitária, mas também desconhecem as boas práticas de preparação de alimentos, constituindo-se em grande risco para a saúde da população. Esse trabalho tem o objetivo de apresentar os resultados da investigação realizada para a identificação de um surto de toxinfecção alimentar ocorrido em um evento científico promovido por uma instituição pública, que contratou um bufê para o fornecimento das refeições. Dentre os 55 participantes que almoçaram no primeiro dia do evento, 28 (51%) adoeceram, tendo sido identificada a Salmonella Typhimurium como causadora do surto e implicado, o sanduíche de tomate seco com queijo branco [TA = 61,5% (24 doentes/39 expostos); RR = 2,46; IC de 95% = 1,02 -5,96; X2 = 4,69; p < 0,05]. Medidas sanitárias foram tomadas a partir da identificação da epidemiologia do surto e do rastreamento dos alimentos visando à prevenção de novos surtos.