Application of flow cytometry for the determination of minimal inhibitory concentration of several antibacterial agents on Mycoplasma hyopneumoniae

Assunção, P.; Antunes, Nuno T.; Rosales, Rubén S.; Poveda, Carlos; Fe, Christian de la; Davey, Hazel M.

doi:10.1111/j.1365-2672.2006.03170.x

Cited by 13 publications

(13 citation statements)

References 30 publications

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“…The results obtained with this approach were comparable with those obtained by the classical broth dilution method, which is dependent on the observation of a color change or increasing turbidity in the growth medium. However, the FCM approach was faster (12 h) than the broth dilution method (48 h), and furthermore, it was concluded that at 48 h FCM was more sensitive for tylosin and at 72 h for oxytetracycline and streptomycin MIC determination [44]. …”

Section: Application Of Flow Cytometrymentioning

confidence: 99%

Applications of Flow Cytometry to Characterize Bacterial Physiological Responses

Ambriz-Aviña

Contreras‐Garduño

Pedraza-Reyes

2014

BioMed Research International

127

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Although reports of flow cytometry (FCM) applied to bacterial analysis are increasing, studies of FCM related to human cells still vastly outnumber other reports. However, current advances in FCM combined with a new generation of cellular reporter probes have made this technique suitable for analyzing physiological responses in bacteria. We review how FCM has been applied to characterize distinct physiological conditions in bacteria including responses to antibiotics and other cytotoxic chemicals and physical factors, pathogen-host interactions, cell differentiation during biofilm formation, and the mechanisms governing development pathways such as sporulation. Since FCM is suitable for performing studies at the single-cell level, we describe how this powerful technique has yielded invaluable information about the heterogeneous distribution of differently and even specialized responding cells and how it may help to provide insights about how cell interaction takes place in complex structures, such as those that prevail in bacterial biofilms.

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Section: Application Of Flow Cytometrymentioning

confidence: 99%

Applications of Flow Cytometry to Characterize Bacterial Physiological Responses

Ambriz-Aviña

Contreras‐Garduño

Pedraza-Reyes

2014

BioMed Research International

127

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“…This is a fast and accurate method for counting the viability of the microorganisms compared to the traditional CFU counting method (23). The use of flow cytometry with DNA dyes and membrane-impermeant probes for determining the viability of various Mycoplasma species has been developed recently (44)(45)(46)(47)(48). We have for the first time applied this method to Ureaplasma spp.…”

Section: Discussionmentioning

confidence: 99%

Suppression of Antimicrobial Peptide Expression by Ureaplasma Species

et al. 2014

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Ureaplasma species commonly colonize the adult urogenital tract and are implicated in invasive diseases of adults and neonates. Factors that permit the organisms to cause chronic colonization or infection are poorly understood. We sought to investigate whether host innate immune responses, specifically, antimicrobial peptides (AMPs), are involved in determining the outcome of Ureaplasma infections. THP-1 cells, a human monocytoid tumor line, were cocultured with Ureaplasma parvum and U. urealyticum. Gene expression levels of a variety of host defense genes were quantified by real-time PCR. In vitro antimicrobial activities of synthetic AMPs against Ureaplasma spp. were determined using a flow cytometry-based assay. Chromosomal histone modifications in host defense gene promoters were tested by chromatin immunoprecipitation (ChIP). DNA methylation status in the AMP promoter regions was also investigated. After stimulation with U. parvum and U. urealyticum, the expression of cell defense genes, including the AMP genes (DEFB1, DEFA5, DEFA6, and CAMP), was significantly downregulated compared to that of TNFA and IL-8, which were upregulated. In vitro flow cytometry-based antimicrobial assay revealed that synthetic peptides LL-37, hBD-3, and hBD-1 had activity against Ureaplasma spp. Downregulation of the AMP genes was associated with chromatin modification alterations, including the significantly decreased histone H3K9 acetylation with U. parvum infection. No DNA methylation status changes were detected upon Ureaplasma infection. In conclusion, AMPs have in vitro activity against Ureaplasma spp., and suppression of AMP expression might be important for the organisms to avoid this aspect of the host innate immune response and to establish chronic infection and colonization.

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“…The application of FC to research and routine procedures [52,193,194] have shown several advantages in diagnostic and the identification of clinically important species [195][196][197], the assessment of drugs and antibiotics effects on cell viability [32,100,130,149,[198][199][200][201] as well as in cell characterization [202,203] and the study of gene regulation and expression in pathogens [98], among others. Recently, the application of an improved two-colour FC assay combining bioconjugated fluorescent nanoparticles and SYBR Green I allowed the detection of Mycobacterium tuberculosis cells avoiding false positives in clinical diagnosis [204].…”

Section: Pharmaceutical Industry and Medical Applicationsmentioning

confidence: 99%