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1.
Microbiol Spectr ; 12(5): e0004724, 2024 May 02.
Article in English | MEDLINE | ID: mdl-38546218

ABSTRACT

Surface waters are considered ecological habitats where Salmonella enterica can persist and disseminate to fresh produce production systems. This study aimed to explore the genomic profiles of S. enterica serotypes Typhimurium, Newport, and Infantis from surface waters in Chile, Mexico, and Brazil collected between 2019 and 2022. We analyzed the whole genomes of 106 S. Typhimurium, 161 S. Newport, and 113 S. Infantis isolates. Our phylogenetic analysis exhibited distinct groupings of isolates by their respective countries except for a notable case involving a Chilean S. Newport isolate closely related to two Mexican isolates, showing 4 and 13 single nucleotide polymorphisms of difference, respectively. The patterns of the most frequently detected antimicrobial resistance genes varied across countries and serotypes. A strong correlation existed between integron carriage and genotypic multidrug resistance (MDR) across serotypes in Chile and Mexico (R > 0.90, P < 0.01), while integron(s) were not detected in any of the Brazilian isolates. By contrast, we did not identify any strong correlation between plasmid carriage and genotypic MDR across diverse countries and serotypes.IMPORTANCEUnveiling the genomic landscape of S. enterica in Latin American surface waters is pivotal for ensuring public health. This investigation sheds light on the intricate genomic diversity of S. enterica in surface waters across Chile, Mexico, and Brazil. Our research also addresses critical knowledge gaps, pioneering a comprehensive understanding of surface waters as a reservoir for multidrug-resistant S. enterica. By integrating our understanding of integron carriage as biomarkers into broader MDR control strategies, we can also work toward targeted interventions that mitigate the emergence and dissemination of MDR in S. enterica in surface waters. Given its potential implications for food safety, this study emphasizes the critical need for informed policies and collaborative initiatives to address the risks associated with S. enterica in surface waters.


Subject(s)
Drug Resistance, Multiple, Bacterial , Phylogeny , Salmonella enterica , Salmonella typhimurium , Serogroup , Salmonella enterica/genetics , Salmonella enterica/isolation & purification , Salmonella enterica/classification , Salmonella enterica/drug effects , Brazil , Drug Resistance, Multiple, Bacterial/genetics , Mexico , Salmonella typhimurium/genetics , Salmonella typhimurium/isolation & purification , Salmonella typhimurium/drug effects , Salmonella typhimurium/classification , Integrons/genetics , Genome, Bacterial , Chile , Genomics , Anti-Bacterial Agents/pharmacology , Latin America , Water Microbiology , Polymorphism, Single Nucleotide , Plasmids/genetics , Microbial Sensitivity Tests
2.
Infect Genet Evol ; 93: 104977, 2021 09.
Article in English | MEDLINE | ID: mdl-34174480

ABSTRACT

Salmonella Typhimurium has been transmitted between humans and animals. Although, Brazil has been one of the largest pork meat exporters worldwide, there are few studies that characterized epidemiologically S. Typhimurium strains from swine. The aims of this work were to study the phylogenetic relationship of S. Typhimurium genomes isolated from swine in Brazil among themselves and with other genomes isolated from several sources and countries using wgMLST and cgMLST and to perform the search of Salmonella pathogenicity islands (SPIs). In addition, for S. Typhimurium strains from swine to compare the virulence and antimicrobial resistance genes by VFDB and ResFinder, genetic content by BLAST Atlas and orthologous proteins clusters by OrthoVenn. The constructed phylogenetic trees by wgMLST and cgMLST grouped the majority (92.3% and 80.7%, respectively) of the strains isolated from swine in Brazil into the same group. All the isolates contained important SPIs (SPI-1, SPI-2, SPI-3, SPI-5 and SPI-9). A total of 100 and 31 virulence and resistance genes were detected in the S. Typhimurium strains isolated from swine, respectively. The BLAST Atlas and orthologous proteins analysis found regions of phages and differences in metabolic, regulatory and cellular processes among S. Typhimurium LT2 and S. Typhimurium isolates from swine. In conclusion, molecular typing based in the wgMLST and cgMLST suggested that the S. Typhimurium isolates from swine studied were genetically related. The pathogenic potential of the strains studied was corroborated by the presence of important SPIs and virulence genes. The high number of antimicrobial resistance genes detected is worrying and reinforced their potential risk in swine in Brazil. The comparison by BLAST Atlas suggested differences in mobile genetic elements among S. Typhimurium LT2 and S. Typhimurium isolates from swine in Brazil. The orthologous proteins analysis revealed unique genes related to important cellular processes in the strains from swine.


Subject(s)
Drug Resistance, Bacterial/genetics , Salmonella Infections, Animal/microbiology , Salmonella typhimurium/genetics , Swine Diseases/microbiology , Animals , Genomic Islands , Molecular Typing , Phylogeny , Salmonella typhimurium/classification , Salmonella typhimurium/pathogenicity , Sus scrofa , Swine , Virulence/genetics
3.
PLoS One ; 15(10): e0240978, 2020.
Article in English | MEDLINE | ID: mdl-33125394

ABSTRACT

International lineages, such as Salmonella Typhimurium sequence type (ST) 19, are most often associated with foodborne diseases and deaths in humans. In this study, we compared the whole-genome sequences of five S. Typhimurium strains belonging to ST19 recovered from clinical human stool samples in North Carolina, United States. Overall, S. Typhimurium strains displayed multidrug-resistant profile, being resistance to critically and highly important antimicrobials including ampicillin, ticarcillin/clavulanic acid, streptomycin and sulfisoxazole, chloramphenicol, tetracycline, respectively. Interestingly, all S. Typhimurium strains carried class 1 integron (intl1) and we were able to describe two genomic regions surrounding blaCARB-2 gene, size 4,062 bp and 4,422 bp for S. Typhimurium strains (HS5344, HS5437, and HS5478) and (HS5302 and HS5368), respectively. Genomic analysis for antimicrobial resistome confirmed the presence of clinically important genes, including blaCARB-2, aac(6')-Iaa, aadA2b, sul1, tetG, floR, and biocide resistance genes (qacEΔ1). S. Typhimurium strains harbored IncFIB plasmid containing spvRABCD operon, as well as rck and pef virulence genes, which constitute an important apparatus for spreading the virulence plasmid. In addition, we identified several virulence genes, chromosomally located, while the phylogenetic analysis revealed clonal relatedness among these strains with S. enterica isolated from human and non-human sources obtained in European and Asian countries. Our results provide new insights into this unusual class 1 integron in virulent S. Typhimurium strains that harbors a pool of genes acting as potential hotspots for horizontal gene transfer providing readily adaptation to new surrounds, as well as being crucially required for virulence in vivo. Therefore, continuous genomic surveillance is an important tool for safeguarding human health.


Subject(s)
Drug Resistance, Multiple, Bacterial , Salmonella Infections/microbiology , Salmonella typhimurium/classification , Virulence Factors/genetics , beta-Lactamases/genetics , Anti-Bacterial Agents/pharmacology , Genome, Bacterial , Humans , Integrons , Microbial Sensitivity Tests , Phylogeny , Plasmids/genetics , Salmonella typhimurium/genetics , Salmonella typhimurium/isolation & purification , Salmonella typhimurium/pathogenicity , United States , Whole Genome Sequencing
4.
Foodborne Pathog Dis ; 17(3): 202-208, 2020 03.
Article in English | MEDLINE | ID: mdl-31556704

ABSTRACT

The mcr-1 gene has been identified in bacterial isolates obtained from humans, animals, environment, and food, including Salmonella spp., which is one of the major foodborne pathogens worldwide. The aim of this study was to evaluate the presence of mcr-1 gene in Salmonella spp. from food produced in Brazil and to characterize the isolates harboring this gene. A total of 490 Salmonella spp. isolates from the Brazilian National Program for the Control of Foodborne Pathogens were screened for the presence of mcr-1 gene by polymerase chain reaction (PCR). Whole genome sequencing (WGS) was performed in positive isolates to characterize the sequence type (ST), plasmid families and resistance genes. Antimicrobial susceptibility tests were performed by broth microdilution. Selected isolates were submitted to conjugation experiments using the Escherichia coli J53 as a receptor. We detected eight isolates harboring the mcr-1 gene; seven belonged to Salmonella enterica serovar Typhimurium and its monophasic variant 4,[5],12:i:-, and one belonged to serovar Saintpaul. Seven of the mcr-1 positive isolates displayed a high rate of resistance to other antibiotics in addition to colistin. Analysis of the WGS indicated that the ST 19 was the most common ST among the mcr-1 positive isolates. The mcr-1 gene was located in an IncX4 plasmid of ∼33 kb, with no additional resistance genes and with high identity with a plasmid obtained from a clinical isolate of E. coli mcr-1 positive in Brazil. All plasmids harboring the mcr-1 gene were able to conjugate. Our results suggest the spread of a single plasmid type in Brazil harboring the mcr-1 among Salmonella spp. The horizontal transfer of this mobile element has been contributing to the spread of the colistin resistance in the country.


Subject(s)
Food Contamination , Food Microbiology , Interspersed Repetitive Sequences , Salmonella enterica/classification , Salmonella enterica/drug effects , Animals , Anti-Bacterial Agents/pharmacology , Brazil/epidemiology , Conjugation, Genetic , DNA, Bacterial , Drug Resistance, Bacterial , Escherichia coli/genetics , Food Microbiology/methods , Genes, Bacterial , Microbial Sensitivity Tests , Phylogeny , Polymerase Chain Reaction , Pork Meat/microbiology , Poultry/microbiology , Salmonella enterica/isolation & purification , Salmonella typhimurium/classification , Salmonella typhimurium/drug effects , Salmonella typhimurium/isolation & purification , Turkeys/microbiology , Whole Genome Sequencing
5.
Braz J Microbiol ; 51(1): 53-64, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31728978

ABSTRACT

Salmonella Typhimurium sequence type 313 (S. Typhimurium ST313) has caused invasive disease mainly in sub-Saharan Africa. In Brazil, ST313 strains have been recently described, and there is a lack of studies that assessed by whole genome sequencing (WGS)-the relationship of these strains. The aims of this work were to study the phylogenetic relationship of 70 S. Typhimurium genomes comparing strains of ST313 (n = 9) isolated from humans and food in Brazil among themselves, with other STs isolated in this country (n = 31) and in other parts of the globe (n = 30) by 16S rRNA sequences, the Gegenees software, whole genome multilocus sequence typing (wgMLST), and average nucleotide identity (ANI) for the genomes of ST313. Additionally, pangenome analysis was performed to verify the heterogeneity of these genomes. The phylogenetic analyses showed that the ST313 genomes were very similar among themselves. However, the ST313 genomes were usually clustered more distantly to other STs of strains isolated in Brazil and in other parts of the world. By pangenome calculation, the core genome was 2,880 CDSs and 4,171 CDSs singletons for all the 70 S. Typhimurium genomes studied. Considering the 10 ST313 genomes analyzed the core genome was 4,112 CDSs and 76 CDSs singletons. In conclusion, the ST313 genomes from Brazil showed a high similarity among them which information might eventually help in the development of vaccines and antibiotics. The pangenome analysis showed that the S. Typhimurium genomes studied presented an open pangenome, but specifically tending to become close for the ST313 strains.


Subject(s)
Food Microbiology , Genome, Bacterial , Phylogeny , Salmonella typhimurium/classification , Bacterial Typing Techniques , Brazil , Feces/microbiology , Genomics , Genotype , Humans , Multilocus Sequence Typing , RNA, Ribosomal, 16S/genetics , Salmonella Infections/microbiology , Virulence Factors/genetics , Whole Genome Sequencing
7.
Braz. J. Pharm. Sci. (Online) ; 56: e18654, 2020. tab, graf
Article in English | LILACS | ID: biblio-1132041

ABSTRACT

The 4-Hydroxycoumarin derivatives are known to show a broad spectrum of pharmacological applications. In this paper we are reporting the synthesis of a new series of 4-Hydroxycoumarin derivatives synthesized through Knovenegal condensation; they were characterized by using UV-Vis, FT-IR, NMR spectroscopies. The synthesized compounds were evaluated for antibacterial activity against Staphylococcus aureus and Salmonella typhimurium strains. The compounds (2), (3) and (8) showed favorable antibacterial activity with zone of inhibitions 26.5± 0.84, 26.0 ± 0.56 and 26.0 ± 0.26 against Staphylococcus aureus (Gram-positive) respectively. However, the compounds (5) and (9) were found more active with 19.5 ± 0.59 and 19.5 ± 0.32 zone of inhibitions against Salmonella typhimurium (Gram-negative). Whereas, in urease inhibition assay, none of the synthesized derivatives showed significant anti-urease activity; although, in carbonic anhydrase-II inhibition assay, the compound (2) and (6) showed enzyme inhibition activity with IC50 values 263±0.3 and 456±0.1, respectively.


Subject(s)
Carbonic Anhydrases/adverse effects , Inhibitory Concentration 50 , Salmonella typhimurium/classification , Urease/adverse effects , Magnetic Resonance Spectroscopy/methods , Condensation
8.
Braz. J. Pharm. Sci. (Online) ; 56: e18468, 2020. tab
Article in English | LILACS | ID: biblio-1249147

ABSTRACT

Origanum vulgare L. (OVEO) essential oil has been considered a candidate antimicrobial for use in food conservation systems. However, studies on the influence of concomitant variations of different food components or physicochemical parameters on the antibacterial properties of OVEO are scarce. This study assessed the influence of concomitant variations in amounts of proteins - PTN (4.0, 6.0 or 8.0 g/100 mL) and lipids - LIP (3.75, 5.0 or 6.25 g/100 mL) and pH values (5.0, 5.5 or 6.0) in cultivation medium on the inhibitory effects of OVEO against Escherichia coli (EC) and Salmonella Typhimurium (ST). Lowest minimum inhibitory concentration values of OVEO against EC and ST were observed in media with the highest LIP amounts regardless the PTN amount and pH value. In absorbance based microtiter plate assay (MPA), for both EC and ST, OVEO caused the lowest Grmax values in medium containing the highest LIP and PTN amounts and lowest pH value. Highest Grmax values for EC and ST were observed in medium containing the lowest LIP and PTN amount and highest pH value. Grmax values estimated from viable counts of EC and ST in tested media with OVEO confirmed bacterial growth behavior similar to that observed in MPA. Overall, the LIP amount in media was as the most influential factor to enhance the antibacterial effects of OVEO. These results indicate that the concomitant influence of LIP and PTN amounts and pH values on the antibacterial effects of OVEO should be considered for optimizing its antimicrobial efficacy in foods.


Subject(s)
Salmonella typhimurium/classification , Oils, Volatile/analysis , Origanum/classification , Escherichia coli/classification , Lipids/adverse effects , Proteins , Microbial Sensitivity Tests/instrumentation , Bacterial Growth , Efficacy , Food , Hydrogen-Ion Concentration
9.
Vet Microbiol ; 231: 147-153, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30955802

ABSTRACT

Salmonella serovars Derby, Typhimurium and the monophasic variant of Salmonella Typhimurium are the most frequently isolated serovars in pigs in France. To compare the excretion patterns, seroconversion to Salmonella and contamination of the organs of pigs inoculated with strains of all three serovars, we conducted an experimental trial with 28 SPF piglets. Four were used as a negative control, while the other 24 were divided equally into three groups. Each group was inoculated at 7 weeks of age with a different strain: S. Derby (SDb), S. Typhimurium (ST), and the monophasic variant of S. Typhimurium (mST). Fecal and blood samples were collected twice a week up until necropsy, on 21 days post-inoculation (DPI) for half of each group and 49 DPI for the remaining piglets. During necropsy, the tonsils, mesenteric lymph nodes and various intestinal contents were collected from each pig. Salmonella bacteria were quantified in CFU/g by a bacteriological method, and levels of Salmonella antibodies were measured using an ELISA Kit. Piglets inoculated with mST continuously excreted Salmonella in their feces throughout the trial. For each of the other serovars, one piglet was Salmonella-negative on one DPI. The quantity of Salmonella excreted was statistically different between the group inoculated with ST and mST (p < 0.05), but no differences were found between the other serovars. The tonsils, cecum and jejunum were the most contaminated organs in all groups. Seroconversion for all the piglets was completed by different DPI: 28 for ST, 31 for mST and 38 for SDb. No major differences were found in terms of excretion and colonization among the studied serovars.


Subject(s)
Antibodies, Bacterial/blood , Salmonella Infections, Animal/immunology , Salmonella Infections, Animal/microbiology , Salmonella typhimurium/classification , Salmonella/classification , Animals , Bacterial Shedding , Cecum/microbiology , Colony Count, Microbial , Enzyme-Linked Immunosorbent Assay , Feces/microbiology , Lymph Nodes/microbiology , Serogroup , Serologic Tests , Swine , Swine Diseases/immunology , Swine Diseases/microbiology
10.
Braz J Microbiol ; 50(2): 539-546, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30680603

ABSTRACT

Macrophages are critical mediators of the inflammatory process, playing a relevant role in the pathogenesis of Salmonella Typhimurium. The protocols for isolation, culture, and differentiation of monocytes into macrophages and their interaction with Salmonella are well established in humans and murine models, but little information is available in swine. The aims of this study were to establish an efficient protocol for macrophage culture and to evaluate the interaction of the invA mutant strain and the wild type (WT) Salmonella Typhimurium with porcine macrophages. Peripheral blood monocyte-derived macrophages from pigs were obtained, separated by density-gradient centrifugation, and cultured in Teflon vials for 10 days. After the differentiation period, cultures consisted of 92.4% CD14+ cells. In addition, these cells showed phagocytic ability, demonstrated by the presence of the same amount of WT and invA mutant Salmonella Typhimurium 1 h after interaction with macrophages. The early cytotoxic effect was Salmonella pathogenicity island (SPI)-[1]dependent, in which log-phase WT strains were more efficient (p < 0.01) than the invA mutant strain at inducing the death of macrophages.


Subject(s)
Macrophages/microbiology , Salmonella Infections, Animal/pathology , Salmonella typhimurium/growth & development , Salmonella typhimurium/pathogenicity , Animals , Bacterial Proteins/genetics , Cells, Cultured , Female , Genomic Islands/genetics , Salmonella typhimurium/classification , Salmonella typhimurium/genetics , Swine , Virulence
11.
Lett Appl Microbiol ; 66(4): 352-359, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29397031

ABSTRACT

Salmonella pathogenicity relies on virulence factors many of which are clustered within the Salmonella pathogenicity islands. Salmonella also harbours mobile genetic elements such as virulence plasmids, prophage-like elements and antimicrobial resistance genes which can contribute to increase its pathogenicity. Here, we have genetically characterized a selected S. Typhimurium strain (CCRJ_26) from our previous study with Multiple Drugs Resistant profile and high-frequency PFGE clonal profile which apparently persists in the pork production centre of Rio de Janeiro State, Brazil. By whole-genome sequencing, we described the strain's genome virulent content and characterized the repertoire of bacterial plasmids, antibiotic resistance genes and prophage-like elements. Here, we have shown evidence that strain CCRJ_26 genome possible represent a virulence-associated phenotype which may be potentially virulent in human infection. SIGNIFICANCE AND IMPACT OF THE STUDY: Whole-genome sequencing technologies are still costly and remain underexplored for applied microbiology in Brazil. Hence, this genomic description of S. Typhimurium strain CCRJ_26 will provide help in future molecular epidemiological studies. The analysis described here reveals a quick and useful pipeline for bacterial virulence characterization using whole-genome sequencing approach.


Subject(s)
Genome, Bacterial/genetics , Salmonella Infections, Animal/microbiology , Salmonella typhimurium/genetics , Swine/microbiology , Animals , Anti-Bacterial Agents , Brazil , Drug Resistance, Bacterial/genetics , Genomic Islands/genetics , Humans , Microbial Sensitivity Tests , Plasmids/genetics , Prophages/genetics , Red Meat/microbiology , Salmonella typhimurium/classification , Salmonella typhimurium/isolation & purification , Virulence/genetics
12.
Int J Med Microbiol ; 308(2): 306-316, 2018 Mar.
Article in English | MEDLINE | ID: mdl-29396155

ABSTRACT

In sub-Saharan Africa, two genetically distinct lineages of multi-drug resistant non-typhoidal Salmonella (NTS) serovar Typhimurium sequence type 313 (ST313) are known to cause invasive disease among people. S. Typhimurium ST313 has evolved to become more human-adapted and is commonly isolated from systemic sites (eg., blood) from febrile patients in sub-Saharan Africa. Epidemiological studies indicate that S. Typhimurium is frequently isolated from systemic sites from human patients in Brazil, however, it is currently unknown if this pathogen has also evolved to become more invasive and human-adapted in this country. Here we determined genotypic and phenotypic divergence among clinical S. Typhimurium strains isolated from systemic and non-systemic sites from human patients in Brazil. We report that a subset (8/38, 20%) of epidemiologically diverse human clinical strains of S. Typhimurium recovered from systemic sites in Brazil show significantly higher intra-macrophage survival, indicating that this subset is likely more invasive. Using the whole genome sequencing and phylogenetic approaches, we identified S. Typhimurium ST313-lineage in Brazil that is genetically and phenotypically distinct from the known African ST313-lineages. We also report the identification of S. Typhimurium ST19-lineage in Brazil that is evolving similar to ST313 lineages from Africa but is genetically and phenotypically distinct from ST19-lineage commonly associated with the gastrointestinal disease worldwide. The identification of new S. Typhimurium ST313 and ST19 lineages responsible for human illnesses in Brazil warrants further epidemiological investigations to determine the incidence and spread of a genetically divergent population of this important human pathogen.


Subject(s)
Adaptation, Physiological/physiology , Salmonella Infections/epidemiology , Salmonella typhimurium/classification , Salmonella typhimurium/genetics , Africa South of the Sahara/epidemiology , Anti-Bacterial Agents/pharmacology , Brazil/epidemiology , Genotype , Humans , Macrophages/microbiology , Microbial Sensitivity Tests , Movement/physiology , Salmonella Infections/microbiology , Salmonella typhimurium/drug effects , Salmonella typhimurium/isolation & purification
13.
Microb Drug Resist ; 23(5): 580-589, 2017 Jul.
Article in English | MEDLINE | ID: mdl-27828759

ABSTRACT

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.


Subject(s)
Drug Resistance, Multiple, Bacterial/genetics , Salmonella Infections/epidemiology , Salmonella typhimurium/genetics , Salmonella/genetics , Serogroup , beta-Lactamases/genetics , Animals , Anti-Bacterial Agents/pharmacology , Brazil/epidemiology , Chickens , Conjugation, Genetic , Electrophoresis, Gel, Pulsed-Field , Gene Expression , Gene Transfer, Horizontal , Genetic Variation , Humans , Isoenzymes/genetics , Isoenzymes/metabolism , Meat Products/microbiology , Microbial Sensitivity Tests , Plasmids/chemistry , Plasmids/metabolism , Prevalence , Public Health Surveillance , Salmonella/classification , Salmonella/drug effects , Salmonella/isolation & purification , Salmonella Infections/drug therapy , Salmonella Infections/microbiology , Salmonella typhimurium/classification , Salmonella typhimurium/drug effects , Salmonella typhimurium/isolation & purification , Sewage/microbiology , beta-Lactamases/metabolism
14.
Biochem Biophys Res Commun ; 470(2): 313-318, 2016 Feb 05.
Article in English | MEDLINE | ID: mdl-26792728

ABSTRACT

Here we present the design of a conditionally lethal mutant of Salmonella enterica serovar Typhimurium (S. Typhimurium) which growth depends on tetracycline (Tet). Four mutants of S. Typhimurium, with Tet-conditional growth, were created by inserting the tetRA cassette. Three of the mutants presented a conditional-lethal phenotype in vitro. One mutant in the yabB gene remained conditional inside cells and did not persisted after 24 h in cell cultures. The capacity of S. Typhimurium yabB::tetRA to invade deep organs was investigated in intraperitoneally (IP) infected mice fed with or without chlortetracycline (CTet), a Tet analog with lower antibiotic activity. The yabB::tetRA mutant was undetectable in liver or spleen of animals under normal diet, while in mice under diet including CTet, yabB::tetRA invaded at a level comparable to the WT in mice under normal diet. Moreover, yabB::tetRA produced a strong humoral-immunoresponse after one IP immunization with 10(6) bacteria, measured as serum reactivity against S. Typhimurium whole cell extract. By contrast, oral immunization with 10(6) bacteria was weaker and variable on inducing antibodies. Consistently, IP infected mice were fully protected in a challenge with 10(4) oral S. Typhimurium, while protection was partial in orally immunized mice. Our data indicate that S. Typhimurium yabB::tetRA is a conditionally attenuated strain capable of inducing a protective response in mice in non-permissive conditions.


Subject(s)
Bacterial Vaccines/administration & dosage , Bacterial Vaccines/microbiology , Salmonella Infections/immunology , Salmonella Infections/prevention & control , Salmonella typhimurium/genetics , Salmonella typhimurium/immunology , Administration, Oral , Animals , Female , Mice , Mice, Inbred BALB C , Mutation/genetics , Salmonella Infections/microbiology , Salmonella typhimurium/classification , Species Specificity
15.
J Med Microbiol ; 64(11): 1395-1407, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26307078

ABSTRACT

Salmonella enterica subsp. enterica serovar Typhimurium is one of the leading serovars that causes salmonellosis worldwide. However, few studies have molecularly characterized S. Typhimurium strains in Brazil. In this study, we genotyped 92 S. Typhimurium strains isolated from humans (43) and food (49) between 1983 and 2013 in Brazil using PFGE, multiple-locus variable number of tandem repeats analysis (MLVA) and enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). Moreover, we assessed the frequency of 12 virulence markers by PCR and the resistance profile against 12 antimicrobials. More than 85.8% of the strains studied carried 11 of the virulence markers or more. Thirty-three strains (25%) were multidrug resistant (MDR). The 92 S. Typhimurium studied were grouped by PFGE as PFGE-A, PFGE-B1 and PFGE-B2; by MLVA as MLVA-A, MLVA-B1 and MLVA-B2; and, finally, by ERIC-PCR as ERIC-A and ERIC-B. The strains isolated from humans before the mid-1990s were allocated to all clusters. The strains isolated from humans after the mid-1990s were distributed in the PFGE-B1, MLVA-B1, MLVA-B2 and ERIC-A clusters. The strains isolated from food were distributed in all clusters, except in PFGE-B2. All typing results suggested that the S. Typhimurium strains of human clinical origin isolated before the mid-1990s were genetically more diverse, which might indicate the selection of a more adapted S. Typhimurium subtype after Salmonella Enteritidis became the most prevalent serovar in Brazil. Regarding strains isolated from food, the results suggest the current circulation of more than one subtype. Furthermore, the high frequency of virulence genes and the presence of MDR strains reinforces their potential hazard for humans and the risk of their presence in foods in Brazil.


Subject(s)
Food Microbiology , Genetic Variation , Salmonella Infections/microbiology , Salmonella typhimurium/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Brazil , Drug Resistance, Bacterial , Food Contamination/analysis , Genotype , Humans , Minisatellite Repeats , Salmonella typhimurium/classification , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics , Virulence Factors/genetics , Virulence Factors/metabolism
16.
J Sci Food Agric ; 94(13): 2705-11, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24676688

ABSTRACT

BACKGROUND: Unpasteurized juice has been associated with foodborne illness outbreaks for many years. Beetroot is a vegetable grown all over the world in temperate areas. In Mexico beetroot is consumed cooked in salads or raw as fresh unpasteurized juices. No data about the microbiological quality or safety of unpasteurized beetroot juices are available. Indicator bacteria, diarrheagenic Escherichia coli pathotypes (DEP) and Salmonella frequencies were determined for fresh unpasteurized beetroot juice from restaurants. RESULTS: One hundred unpasteurized beetroot juice samples were collected from public markets in Pachuca, Mexico. Frequencies in these samples were 100%, 75%, 53%, 9% and 4% of positive samples, for coliform bacteria, fecal coliforms, E. coli, DEP and Salmonella, respectively. Identified DEP included enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC) and non-O157 Shiga toxin-producing E. coli (STEC). Identified Salmonella serotypes included Typhimurium and Enteritidis. CONCLUSION: This is the first report of microbiological quality and atypical EPEC, ETEC, non-O157 STEC and Salmonella isolation from fresh raw beetroot juice in Mexico. Fresh raw beetroot juice from markets is very probably an important factor contributing to the endemicity of atypical EPEC, ETEC, non-O157 STEC and Salmonella-related gastroenteritis in Mexico.


Subject(s)
Beta vulgaris/microbiology , Beverages/microbiology , Enteropathogenic Escherichia coli/growth & development , Enterotoxigenic Escherichia coli/growth & development , Plant Roots/microbiology , Salmonella/growth & development , Shiga-Toxigenic Escherichia coli/growth & development , Beverages/economics , Colony Count, Microbial , Enteropathogenic Escherichia coli/classification , Enteropathogenic Escherichia coli/isolation & purification , Enterotoxigenic Escherichia coli/classification , Enterotoxigenic Escherichia coli/isolation & purification , Mexico , Molecular Typing , Multiplex Polymerase Chain Reaction , Restaurants , Salmonella/classification , Salmonella/isolation & purification , Salmonella enteritidis/classification , Salmonella enteritidis/growth & development , Salmonella enteritidis/isolation & purification , Salmonella typhimurium/classification , Salmonella typhimurium/growth & development , Salmonella typhimurium/isolation & purification , Shiga-Toxigenic Escherichia coli/classification , Shiga-Toxigenic Escherichia coli/isolation & purification
17.
Clin Infect Dis ; 55(1): 51-60, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22423121

ABSTRACT

BACKGROUND: The foodborne transmission and human health impact of Salmonella and Campylobacter infections have rarely been evaluated at the population level in highly endemic settings. METHODS: A prospective 15-month cohort study of 127 infants and 119 elderly people was combined with animal and food surveillance to determine the incidence and severity of Salmonella and Campylobacter gastroenteritis in a comparatively prosperous rural community in Mexico. RESULTS: Salmonella and Campylobacter were isolated in up to 75% and 57%, respectively, of raw retail meat and in up to 4.5% of ready-to-eat foods. Rates of acute gastroenteritis of any etiology in infants and elderly people were, respectively, 2.1 and 0.7 episodes per person per year. The annual incidence density rate of Salmonella gastroenteritis was 17.8 per 100 infants and 7.9 per 100 elderly people; the rate of Campylobacter gastroenteritis was 11.7 per 100 infants and 0 per 100 elderly people. Pulsed-field gel electrophoresis analysis yielded multiple clusters of human, meat, and/or animal Salmonella and Campylobacter isolates with indistinguishable patterns. On average, gastroenteritis episodes with these pathogens lasted 3 days in infants and 2 days in elderly people. Medical attention was sought in 44% of diarrheal episodes in infants and in 26% of diarrheal episodes in elderly people; none required hospitalization. Infants with multidrug-resistant Salmonella gastroenteritis had a higher frequency of bloody stools and medical visits (50% vs 11%; odds ratio, 8.5; P = .04) than those with more susceptible strains. CONCLUSIONS: In this relatively advantaged Mexican rural community, the human health impact of a food chain heavily contaminated with Salmonella and Campylobacter was of low magnitude.


Subject(s)
Campylobacter Infections/epidemiology , Disease Transmission, Infectious/statistics & numerical data , Rural Population/statistics & numerical data , Salmonella Food Poisoning/epidemiology , Aged , Aged, 80 and over , Animals , Anti-Bacterial Agents/pharmacology , Campylobacter Infections/microbiology , Campylobacter jejuni/classification , Campylobacter jejuni/genetics , Campylobacter jejuni/isolation & purification , Cattle , Chi-Square Distribution , Cluster Analysis , Electrophoresis, Gel, Pulsed-Field , Endemic Diseases , Female , Food Chain , Gastroenteritis/epidemiology , Gastroenteritis/microbiology , Humans , Infant , Male , Meat/microbiology , Mexico/epidemiology , Microbial Sensitivity Tests , Prevalence , Prospective Studies , Salmonella Food Poisoning/microbiology , Salmonella Infections, Animal/epidemiology , Salmonella Infections, Animal/microbiology , Salmonella typhimurium/classification , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics , Salmonella typhimurium/isolation & purification , Socioeconomic Factors , Swine , Zoonoses
18.
Rev Panam Salud Publica ; 29(6): 387-92, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21829960

ABSTRACT

OBJECTIVE: To determine the prevalence of resistance to ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracyclines (ACSSuT) in Salmonella serovar Typhimurium definitive [phage] type (DT) 193 strains isolated from human sources over the last four decades. METHODS: From 2008 to 2010, 553 DT193 isolates out of 810 human-origin Salmonella ser. Typhimurium phage-typed strains isolated from the 1970s through 2008 were selected and tested for ACSSuT resistance: 91 strains isolated during the 1970s, 65 from the 1980s, 70 from the 1990s, and 327 from 2000-2008. Resistance profiles were determined using the disk diffusion method. RESULTS: An antimicrobial susceptibility assay indicated 20.9%, or 116, of all isolates tested were ACSSuT-resistant, 52.0% (287) were resistant to one or more drugs in the ACSSuT profile, and 27.1% (150) were nonresistant (susceptible to antimicrobials). Based on the assay, overall antimicrobial resistance was extremely high in the 1970s (affecting 99.0% of isolates from that period) and remained high during the 1980s, when 95.4% of isolates had some type of antimicrobial resistance and incidence of Salmonella ser. Typhimurium DT193 R-type ACSSuT increased to 73.8%. R-type ACSSuT dropped to 27.1% (19 isolates) during the 1990s, and to 5.2% (17) during 2000-2008, despite a substantial increase in the number of isolates tested (397 versus 204, 111, and 98, respectively, for the previous three decades). CONCLUSIONS: Although prevalence of Salmonella ser. Typhimurium DT193 R-type ACSSuT in Brazil has decreased since the 1970s, ACSSuT resistance markers continue to circulate. Therefore, continuous surveillance should be conducted to evaluate the occurrence of Salmonella ser. Typhimurium DT193 and its antimicrobial resistance.


Subject(s)
Drug Resistance, Multiple, Bacterial , R Factors/genetics , Salmonella Infections/microbiology , Salmonella typhimurium/drug effects , Animals , Anti-Bacterial Agents/pharmacology , Bacteriophage Typing , Brazil/epidemiology , Drug Resistance, Multiple, Bacterial/genetics , Food Chain , Humans , Retrospective Studies , Salmonella Infections/drug therapy , Salmonella Infections/epidemiology , Salmonella Infections, Animal/microbiology , Salmonella typhimurium/classification , Salmonella typhimurium/genetics , Salmonella typhimurium/isolation & purification , Serotyping , Zoonoses
19.
Rev. panam. salud pública ; 29(6): 387-392, June 2011. tab
Article in English | LILACS | ID: lil-608267

ABSTRACT

OBJECTIVE: To determine the prevalence of resistance to ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracyclines (ACSSuT) in Salmonella serovar Typhimurium definitive [phage] type (DT) 193 strains isolated from human sources over the last four decades. METHODS: From 2008 to 2010, 553 DT193 isolates out of 810 human-origin Salmonella ser. Typhimurium phage-typed strains isolated from the 1970s through 2008 were selected and tested for ACSSuT resistance: 91 strains isolated during the 1970s, 65 from the 1980s, 70 from the 1990s, and 327 from 2000-2008. Resistance profiles were determined using the disk diffusion method. RESULTS: †An antimicrobial susceptibility assay indicated 20.9 percent, or 116, of all isolates tested were ACSSuT-resistant, 52.0 percent (287) were resistant to one or more drugs in the ACSSuT profile, and 27.1 percent (150) were nonresistant (susceptible to antimicrobials). Based on the assay, overall antimicrobial resistance was extremely high in the 1970s (affecting 99.0 percent of isolates from that period) and remained high during the 1980s, when 95.4 percent of isolates had some type of antimicrobial resistance and incidence of Salmonella ser. Typhimurium DT193 R-type ACSSuT increased to 73.8 percent. R-type ACSSuT dropped to 27.1 percent (19 isolates) during the 1990s, and to 5.2 percent (17) during 2000-2008, despite a substantial increase in the number of isolates tested (397 versus 204, 111, and 98, respectively, for the previous three decades). CONCLUSIONS: †Although prevalence of Salmonella ser. Typhimurium DT193 R-type ACSSuT in Brazil has decreased since the 1970s, ACSSuT resistance markers continue to circulate. Therefore, continuous surveillance should be conducted to evaluate the occurrence of Salmonella ser. Typhimurium DT193 and its antimicrobial resistance.


OBJETIVO: Determinar la prevalencia de resistencia a la ampicilina, el cloranfenicol, la estreptomicina, las sulfonamidas y las tetraciclinas (ACSSuT) en cepas de Salmonella serovariedad Typhimurium fagotipo definitivo (DT) 193 aisladas de fuentes de origen humano durante las cuatro últimas décadas. MÉTODOS: Entre el 2008 y el 2010 se seleccionaron 553 aislados de DT193 entre 810 cepas de Salmonella serovariedad Typhimurium fagotipificadas aisladas desde la década de 1970 hasta el 2008, y en ellos se analizó la resistencia a ACSSuT: se estudiaron 91 cepas aisladas durante la década de 1970, 65 aisladas durante la década de 1980, 70 aisladas durante la década de 1990, y 327 aisladas entre el 2000 y el 2008, respectivamente. Los perfiles de resistencia a los antibióticos se determinaron mediante el método de difusión en disco. RESULTADOS: El antibiograma indicó que 20,9 por ciento (116) de todos los aislados que se analizaron fueron resistentes a ACSSuT, 52,0 por ciento (287) fueron resistentes a uno o más antibióticos del grupo ACSSuT y 27,1 por ciento (150) no fueron resistentes (es decir, fueron sensibles a dichos antibióticos). Según el análisis, la resistencia general a los antibióticos fue muy alta en la década de 1970 (y comprendió a 99,0 por ciento de los aislados de ese período) y continuó siendo alta durante la década de 1980, cuando 95,4 por ciento de los aislados presentó algún tipo de resistencia a los antibióticos y la incidencia de Salmonella serovariedad Typhimurium DT193 con resistencia de tipo ACSSuT aumentó hasta 73,8 por ciento. La resistencia de tipo ACSSuT descendió a 27,1 por ciento (31 aislados) durante la década de 1990, y a 5,2 por ciento (17 aislados) entre el 2000 y el 2008, a pesar del aumento importante en el número de aislados que se evaluaron (397 frente a 204, 111 y 98 en las tres décadas anteriores, respectivamente). CONCLUSIONES: Aunque la prevalencia de Salmonella serovariedad Typhimurium DT193 con resistencia de tipo ACSSuT en el Brasil ha disminuido desde la década de 1970, los marcadores de resistencia a ACSSuT continúan en circulación. Por consiguiente, debe llevarse a cabo una vigilancia permanente para evaluar la aparición de infecciones por Salmonella serovariedad Typhimurium DT193 y su resistencia a los antibióticos.


Subject(s)
Humans , Animals , Drug Resistance, Multiple, Bacterial , R Factors/genetics , Salmonella Infections/microbiology , Salmonella typhimurium/drug effects , Anti-Bacterial Agents/pharmacology , Bacteriophage Typing , Brazil/epidemiology , Drug Resistance, Multiple, Bacterial/genetics , Food Chain , Retrospective Studies , Salmonella Infections, Animal/microbiology , Salmonella Infections/drug therapy , Salmonella Infections/epidemiology , Salmonella typhimurium/classification , Salmonella typhimurium/genetics , Salmonella typhimurium/isolation & purification , Serotyping , Zoonoses
20.
Microb Drug Resist ; 16(3): 185-90, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20704512

ABSTRACT

The increase of Salmonella enterica strains showing resistance against antibiotics has resulted in limiting the effective treatment of human infections. The present study characterized the resistance to tetracycline in S. enterica serovar Typhimurium strains, recovered from irrigation water in distinct regions in the Culiacan Valley, an important agricultural region in Mexico for horticultural crops that are exported to the United States. Analysis of the genomic diversity by pulse-field gel electrophoresis (PFGE) typing showed that the Salmonella Typhimurium strains were grouped into four distinct genotypic clusters, indicating genomic diversity among 12 strains examined. The polymerase chain reaction and DNA sequencing analysis demonstrated that the tet(A) gene was found on the genomic DNA and was located within a truncated version of transposon Tn1721. The comparative analysis of the tet(A) gene sequence in Salmonella Typhimurium strains identified high sequence similarity to the tet determinant of plasmid RP1, which is homologous to the tet gene in Tn1721. The findings show the presence of tet(A) among the tetracycline-resistant Salmonella Typhimurium strains isolated from irrigation water used for growing fresh fruits and vegetables.


Subject(s)
Agricultural Irrigation/methods , Antiporters/genetics , Bacterial Proteins/genetics , Fresh Water/microbiology , Salmonella typhimurium/drug effects , Salmonella typhimurium/genetics , Tetracycline Resistance/genetics , Animals , Anti-Bacterial Agents/pharmacology , DNA Transposable Elements , Electrophoresis, Gel, Pulsed-Field , Genotype , Humans , Mexico , Microbial Sensitivity Tests , Phylogeny , Polymerase Chain Reaction , Salmonella typhimurium/classification , Salmonella typhimurium/isolation & purification , Sequence Analysis, DNA
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