In-vitro antimicrobial susceptibility of <i>Yersinia enterocolitica</i> isolates from stools of patients in The Netherlands from 1982–1991

Stolk-Engelaar, Virginia M. M.; Meis, Jacques F.; Mulder, Janet A.; Loeffen, Frans; Hoogkamp-Korstanje, J. A. A.

doi:10.1093/jac/36.5.839

Cited by 22 publications

(12 citation statements)

References 6 publications

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“…Bacterial growth was quantified by absorbance (A 600nm ) and mCherry fluorescence was quantified in a microplate reader. Treatment with 1µg/ml Dox resulted in significant growth inhibition relative to untreated cells, 0.01µg/ml Dox, and 0.1µg/ml Dox (Figure 1B), which is consistent with previously published Dox minimal inhibitory concentrations (MIC) levels for Yersinia (24–26). Reporter signal was detected with 0.01µg/ml Dox, and significantly increased with 0.1µg/ml Dox; neither lower dose inhibited bacterial growth (Figure 1B, 1C).…”

Section: Resultssupporting

confidence: 91%

Subpopulations of stressedY. pseudotuberculosispreferentially survive doxycycline treatment within host tissues

Raneses

Ellison

Liu

et al. 2020

Preprint

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Severe systemic bacterial infections result in colonization of deep tissues, which can be very difficult to eliminate with antibiotics. It remains unclear if this is because antibiotics are not reaching inhibitory concentrations within tissues, if subsets of bacteria are less susceptible to antibiotics, or if both contribute to limited treatment efficacy. To determine the concentration of doxycycline (Dox) present within deep tissues following treatment, we generated a fluorescent transcriptional reporter derived from the tet operon to specifically detect intracellular tetracycline exposure at the single bacterial cell level. Dox exposure was detected in the spleen 2 hours after intraperitoneal injection, and by 4 hours post-injection, this treatment resulted in a significant decrease in viable Yersinia pseudotuberculosis in the spleen. Nitric oxide-stressed bacteria preferentially survived treatment, suggesting stress was sufficient to alter Dox susceptibility. Many bacteria (~10%) survived a single dose of Dox, and the antibiotic accumulated at the periphery of microcolonies to growth inhibitory concentrations until 48 hours post-treatment. After this timepoint, antibiotic concentrations decreased and bacterial growth resumed. Dox-treated mice eventually succumbed to the infection, albeit with significantly prolonged survival relative to untreated mice. These results indicate that Dox delivery by intraperitoneal injection results in rapid diffusion of inhibitory concentrations of antibiotic into the spleen, but stressed cells preferentially survive drug treatment, and bacterial growth resumes once drug concentrations decrease. This fluorescent reporter strategy for antibiotic detection could easily be modified to detect the concentration of additional antimicrobial compounds within host tissues following drug administration.ImportanceBacterial infections are very difficult to treat when bacteria spread into the bloodstream and begin to replicate within deep tissues, such as the spleen. Subsets of bacteria can survive antibiotic treatment, but it remains unclear if this survival is because of limited drug diffusion into tissues, or if something has changed within the bacteria, promoting survival of some bacterial cells. Here, we have developed a fluorescent reporter to detect doxycycline (Dox) diffusion into host tissues, and show that Dox impacts the bacterial population within hours of administration, and inhibits bacterial growth for 48 hours. However, bacterial growth resumes when antibiotic concentrations decrease. Subsets of bacteria, stressed by the host response to infection, survive Dox treatment at a higher rate. These results provide critical information about the dynamics that occur within deep tissues following antibiotic administration, and suggests subsets of bacteria are predisposed to survive inhibitory concentrations of antibiotic before exposure.

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Section: Resultssupporting

confidence: 91%

Subpopulations of stressedY. pseudotuberculosispreferentially survive doxycycline treatment within host tissues

Raneses

Ellison

Liu

et al. 2020

Preprint

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“…Stolk-Engelaar et al examined 335 strains and could not associate susceptibility patterns with biovar or serovar [3], but others have found relationships . Previous studies at this laboratory confirmed the latter view, finding a relationship between biovar and susceptibility to ticarcillin, amoxycillin-clavulanate, cefazolin, loracarbef, cefoxitin, ce$odoxime, cefixime and other p-lactam antibiotics (Table 1 To elucidate the molecular basis for the differences in susceptibilities to p-lactam antibiotics of biovar 2, 4 and 5 strains, their P-lactamases were characterised genotypically and phenotypically.…”

Section: Hares Ismentioning

confidence: 99%

Expression of β-lactamases in Yersinia enterocolitica strains of biovars 2, 4 and 5

Stock¹,

Heisig²,

Wiedemann³

1999

Journal of Medical Microbiology

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Characteristic patterns of susceptibility to p-lactam antibiotics are associated with different biovars of Yersinia enterucditica. To elucidate the basis for these differences, the P-lactamases of strains of Y: enterocditica biovars 4 (n = 63), 2 (n = 12) and 5 (n = 10) were characterised. PCR fragments were generated from the P-lactamase A (bZaA) and B (bZaB) genes; in addition, P-lactamase induction tests were performed with imipenem as the inducer and P-lactamase inhibition assays were undertaken with aztreonam and clavulanic acid. All the strains yielded PCR amplification fragments with primers to bZaA and bZaB. Biovar 4 strains had uniform patterns of P-lactamase induction and inhibition: uninduced biovar 4 strains predominantly expressed BlaA, but low-level expression of BlaB was also detected; after induction, biovar 4 strains predominantly produced BlaB. P-Lactamase expression varied between and within biovars 2 and 5: uninduced strains predominantly expressed either BlaA or BlaB, or exclusively BlaB; after induction BlaB was predominantly or exclusively expressed. Both the basal and induced levels of P-lactamase varied within biovars 2 and 5. Some biovar 5 strains were not inducible; these predominantly produced BlaA. The results of this study show that biovar 2, 4 and 5 strains contain both bZaA and bZaB, but that the expression of the enzymes is regulated differently between the biovars, and varies within biovars 2 and 5.

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“…Stolk-Engelaar et al examined strains of all biovars except biovar 5 and found no association between susceptibility and biovar or serovar [11], whereas others have found relationships [9, 10, 21±23]. This group con®rmed the latter view, ®nding a relationship between biovar and susceptibility to several â-lactam antibiotics (Table 1 and [22]).…”

Section: Introductionmentioning

confidence: 94%

β-Lactamase expression in Yersinia enterocolitica biovars 1A, 1B and 3

Stock

Heisig

Wiedemann

2000

Journal of Medical Microbiology

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In-vitro antimicrobial susceptibility of Yersinia enterocolitica isolates from stools of patients in The Netherlands from 1982–1991

Cited by 22 publications

References 6 publications

Subpopulations of stressedY. pseudotuberculosispreferentially survive doxycycline treatment within host tissues

Subpopulations of stressedY. pseudotuberculosispreferentially survive doxycycline treatment within host tissues

Expression of β-lactamases in Yersinia enterocolitica strains of biovars 2, 4 and 5

β-Lactamase expression in Yersinia enterocolitica biovars 1A, 1B and 3

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