Antimicrobial resistance of Staphylococcus spp. isolated from organic and conventional Minas Frescal cheese producers in São Paulo, Brazil

Abreu, Anderson Clayton da Silva; Matos, Luiz Gustavo de; Cândido, Talita Junia da Silva; Barboza, Giovana Rueda; Souza, Victória Vilaça Martins Alencar de; Nuñez, Karen Vanessa Munive; Silva, Nathália Cristina Cirone

doi:10.3168/jds.2020-19338

Cited by 26 publications

(18 citation statements)

References 60 publications

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“…PAsmr has also been shown to transport methyl viologen and tetraphenylphosphonium (TPP + ) in vitro (Ninio et al., 2001). SAsmr (QacC) from S. aureus has been shown to confer resistance to ethidium bromide in both Staphylococcus and E. coli (Abd El‐Aziz et al., 2021; da Silva Abreu et al., 2021; Fuentes et al., 2005; Lee et al., 2020; Tang et al., 2020) and has previously been expressed and purified at the scale needed for biophysical characterization (Poget et al., 2007, 2010). FTsmr from F. tularensis , the causative agent of tularemia, is almost completely uncharacterized, but an smr sequence has been identified as part of an arsenic resistance locus in Francisella species (Kassinger & van Hoek, 2021; Snowden & Simonsen, 2022).…”

Section: Introductionmentioning

confidence: 99%

Functional promiscuity of small multidrug resistance transporters from Staphylococcus aureus, Pseudomonas aeruginosa, and Francisella tularensis

Spreacker,

Wegrzynowicz,

Porter

et al. 2024

Molecular Microbiology

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Small multidrug resistance transporters efflux toxic compounds from bacteria and are a minimal system to understand multidrug transport. Most previous studies have focused on EmrE, the model SMR from Escherichia coli, finding that EmrE has a broader substrate profile than previously thought and that EmrE may perform multiple types of transport, resulting in substrate‐dependent resistance or susceptibility. Here, we performed a broad screen to identify potential substrates of three other SMRs: PAsmr from Pseudomonas aeruginosa; FTsmr from Francisella tularensis; and SAsmr from Staphylococcus aureus. This screen tested metabolic differences in E. coli expressing each transporter versus an inactive mutant, for a clean comparison of sequence and substrate‐specific differences in transporter function, and identified many substrates for each transporter. In general, resistance compounds were charged, and susceptibility substrates were uncharged, but hydrophobicity was not correlated with phenotype. Two resistance hits and two susceptibility hits were validated via growth assays and IC50 calculations. Susceptibility is proposed to occur via substrate‐gated proton leak, and the addition of bicarbonate antagonizes the susceptibility phenotype, consistent with this hypothesis.

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Section: Introductionmentioning

confidence: 99%

Functional promiscuity of small multidrug resistance transporters from Staphylococcus aureus, Pseudomonas aeruginosa, and Francisella tularensis

Spreacker,

Wegrzynowicz,

Porter

et al. 2024

Molecular Microbiology

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“…Coagulase-negative staphylococci and M. caseolyticus are known to harbor a variety of genes encoding antibiotic resistance, including mecA, mecB, mecC, and mecD (Baba et al, 2009; International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements, 2009;Fontes et al, 2013;Chajęcka-Wierzchowska et al, 2015;Schwendener, et al, 2017;Schwendener, et al, 2019;Becker, 2021;da Silva Abreu et al, 2021). In addition, it has been shown that bacteria are able or at least have the potential to exchange genes with another species of the same genus or even a species of another genus (Alonso et al, 2001;Resch et al, 2008;Becker et al, 2018;Schwendener and Perreten, 2018).…”

Section: Introductionmentioning

confidence: 99%

Characterization of coagulase-negative staphylococci and macrococci isolated from cheese in Germany

Klempt

Franz

Hammer

2022

Journal of Dairy Science

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Cheese, especially ripened varieties, harbor a very complex and heterogeneous microbiota. In addition to the desired microorganisms (starter cultures) added during cheese production, potentially harmful bacteria may also enter the production chain. Regarding the latter, the focus of this study was on coagulase-negative staphylococci (CNS) and Macrococcus caseolyticus. Both are known to harbor a variety of genes coding for antibiotic resistance, including mecA, mecB, mecC, and mecD. Coagulase-negative staphylococci or macrococci carrying such genes or other virulence factors should not be present in cheese. Cheese samples (101 in total) were collected from retail sources. Coagulase-negative staphylococci and M. caseolyticus were isolated utilizing selective agars, and species were identified by phenotypical tests and partial sequencing of the sodA gene. The results allowed identification of 53 CNS strains and 19 M. caseolyticus strains. Among the CNS, 11 isolates of Staphylococcus saprophyticus and one Staphylococcus epidermidis isolate were obtained. Both species are potential human pathogens and may thus adversely affect the safety of these food products. Screening for antimicrobial resistance was performed by application of disc diffusion tests, a gradient strip-test, and 14 different PCR tests. Evidence for methicillin resistance (by either positive disc diffusion assay for cefoxitin or by mec PCR) was found in CNS isolates and M. caseolyticus (9 isolates each). Regarding other virulence factors, no genetic determinants for coagulase or the most common staphylococcal enterotoxins sea, seb, sec, sed, and see were detected in any of the CNS or M. caseolyticus isolates by PCR testing. In conclusion, the presence of facultatively pathogenic CNS and carriers of genes for antibiotic resistance in both groups of microorganisms, especially mec genes, and the respective food safety issues need further evaluation and surveillance.

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“…These studies show that EmrE homologs in other bacterial species can confer resistance to similar substrates, but the scope of the assays was limited to common multidrug resistance substrates and detection of resistance phenotypes. Here we analyze SMRs from three bacterial species of interest for the potential to cause disease in humans or animals for comparison with EmrE: Staphylococcus aureus (SAsmr) (42)(43)(44)(45), Pseudomonas aeruginosa (PAsmr) (46,47), and Francisella tularensis (FTsmr) (48). These particular transporters were selected on the basis of disease relevance, sequence diversity (Fig.…”

Section: Introductionmentioning

confidence: 99%

Functional promiscuity of small multidrug resistance transporters fromStaphylococcus aureus,Pseudomonas aeruginosa, andFrancisella tularensis

Spreacker¹,

Porter²,

Wegrzynowicz³

et al. 2022

Preprint

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Small multidrug resistance (SMR) transporters efflux toxic substrates from bacterial cells and were recently divided into two subfamilies: specific toxic metabolite transporters and promiscuous drug exporters. These drug exporters are thought to function similarly to EmrE, the model system for this subfamily of SMR transporters. Studies of EmrE homologs indicate that they are able to confer resistance to EmrE substrates in E. coli and in their native organisms. Recent work from our lab showed that functional EmrE can confer resistance or susceptibility in vivo depending on the drug substrate. Here, we test whether this functional promiscuity of EmrE extends to SMR transporters from three additional human or animal pathogens: SAsmr from Staphylococcus aureus, PAsmr from Pseudomonas aeruginosa, and FTsmr from Francisella tularensis. We find that these SMR homologs can confer either resistance or susceptibility to different toxic substrates in E. coli. This demonstrates that the ability of a single transporter to lead to opposite biological outcomes when transporting different substrates is a general property of the promiscuous multidrug transporters in the SMR family. It also suggests the potential for novel antibiotic development targeting these transporters with small molecules that trigger susceptibility. Such a strategy does not require that the target be the primary mode for antibiotic resistance because the goal is not simple inhibition of activity, but rather activation of an alternative transport function that is detrimental to bacteria.

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Antimicrobial resistance of Staphylococcus spp. isolated from organic and conventional Minas Frescal cheese producers in São Paulo, Brazil

Cited by 26 publications

References 60 publications

Functional promiscuity of small multidrug resistance transporters from Staphylococcus aureus, Pseudomonas aeruginosa, and Francisella tularensis

Functional promiscuity of small multidrug resistance transporters from Staphylococcus aureus, Pseudomonas aeruginosa, and Francisella tularensis

Characterization of coagulase-negative staphylococci and macrococci isolated from cheese in Germany

Functional promiscuity of small multidrug resistance transporters fromStaphylococcus aureus,Pseudomonas aeruginosa, andFrancisella tularensis

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