Evaluation of Mycoplasma hyopneumoniae growth by flow cytometry

Assunção, P.; Rivera, Díaz; Comas, Jaume; Galarreta, C. M. Ruiz de; González-Llamazares, O. R.

doi:10.1111/j.1365-2672.2005.02536.x

Cited by 17 publications

(20 citation statements)

References 32 publications

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“…2005), the causal agent of porcine enzootic pneumonia, which causes high economic losses in porcine production (Kobisch and Friis 1996). Despite the fact that M. hyopneumoniae strains exhibit a reduced ability to form colonies (Friis 1977), we demonstrated a good relationship between growth estimations performed using the semiquantitative CCU and FC method (Assunção et al. 2005).…”

Section: Introductionmentioning

confidence: 79%

“…According to these considerations we recently used FC (Assunção et al. 2005) for the rapid estimation of M. hyopneumoniae , a pathogen responsible for porcine enzootic pneumonia.…”

Section: Discussionmentioning

confidence: 99%

“…2001). In this context, we recently reported that FC was a suitable method to enumerate Mycoplasma hyopneumoniae (Assunção et al. 2005), the causal agent of porcine enzootic pneumonia, which causes high economic losses in porcine production (Kobisch and Friis 1996).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Use of flow cytometry for enumeration of Mycoplasma mycoides subsp. mycoides large-colony type in broth medium

Assunção

Antunes

et al. 2006

J Appl Microbiol

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Aims: The potential of using flow cytometry (FC) in combination with a fluorescent dye (SYBR green‐I) for rapidly estimating Mycoplasma mycoides subSPS. mycoides large‐colony type (MmmLC) in broth culture was examined. Methods and Results: The FC analysis was performed by staining the MmmLC cells with a fluorescent dye, SYBR green‐I (SYBR), and the results were compared with plate count method (colony forming units, – CFUs). There was a good correlation (linear regression, r2 = 0·93) between mycoplasma counts determined by FC (cells ml−1) and by traditional plate count method (CFU ml−1). The lowest bacterial concentration detected by FC and traditional plate count was of the order of 104 cells ml−1 and 103 CFU ml−1, respectively. FC method allowed results in 20–30 min, whereas at least 24 h were necessary to obtain results with the traditional plate count method (CFU). Conclusion: Growth rates of MmmLC in broth medium determined by FC were highly reproducible and correlated well with mycoplasma counts assessed by the plate count method. Significance and Impact of the Study: These findings suggest that FC could be a good alternative to replace other time‐consuming techniques that are currently used to enumerate mycoplasma in broth medium, such as plate count method (CFU).

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Section: Introductionmentioning

confidence: 79%

“…According to these considerations we recently used FC (Assunção et al. 2005) for the rapid estimation of M. hyopneumoniae , a pathogen responsible for porcine enzootic pneumonia.…”

Section: Discussionmentioning

confidence: 99%

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Use of flow cytometry for enumeration of Mycoplasma mycoides subsp. mycoides large-colony type in broth medium

Assunção

Antunes

et al. 2006

J Appl Microbiol

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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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“…M. hyopneumoniae non-pathogenic strain J (ATCC 25,934), and the non-pathogenic M. flocculare (ATCC 27,716) were acquired from American Type Culture Collection by the Empresa Brasileira de Pesquisa Agropecuária-Centro Nacional de Pesquisa de Suínos e Aves (EMBRAPA-CNPSA, Concórdia, SC, Brazil). For soluble and insoluble protein extracts, respectively, all bacteria were cultivated in 50 mL and 100 mL of Friis medium [44] for 48 h [45], at 37°C. Cultures were carried out independently in triplicates (biological replicates), and immediately used for protein extraction.…”

Section: Methodsmentioning

confidence: 99%

Comparative proteomics of twoMycoplasma hyopneumoniaestrains andMycoplasma flocculareidentified potential porcine enzootic pneumonia determinants

et al. 2018

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Mycoplasma hyopneumoniae and Mycoplasma flocculare are genetically similar bacteria, which coinhabit the porcine respiratory tract. These mycoplasmas share most of the known virulence factors, but, while M. hyopneumoniae causes porcine enzootic pneumonia (PEP), M. flocculare is a commensal species. To identify potential PEP determinants and provide novel insights on mycoplasma-host interactions, the whole cell proteomes of two M. hyopneumoniae strains, one pathogenic (7448) and other non-pathogenic (J), and M. flocculare were compared. A cell fractioning approach combined with mass spectrometry (LC-MS/MS) proteomics was used to analyze cytoplasmic and surface-enriched protein fractions. Average detection of ~ 50% of the predicted proteomes of M. hyopneumoniae 7448 and J, and M. flocculare was achieved. Many of the identified proteins were differentially represented in M. hyopneumoniae 7448 in comparison to M. hyopneumoniae J and M. flocculare, including potential PEP determinants, such as adhesins, proteases, and redox-balancing proteins, among others. The LC-MS/MS data also provided experimental validation for several genes previously regarded as hypothetical for all analyzed mycoplasmas, including some coding for proteins bearing virulence-related functional domains. The comprehensive proteome profiling of two M. hyopneumoniae strains and M. flocculare provided tens of novel candidates to PEP determinants or virulence factors, beyond those classically described.

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Evaluation of Mycoplasma hyopneumoniae growth by flow cytometry

Cited by 17 publications

References 32 publications

Use of flow cytometry for enumeration of Mycoplasma mycoides subsp. mycoides large-colony type in broth medium

Use of flow cytometry for enumeration of Mycoplasma mycoides subsp. mycoides large-colony type in broth medium

Suppression of Antimicrobial Peptide Expression by Ureaplasma Species

Comparative proteomics of twoMycoplasma hyopneumoniaestrains andMycoplasma flocculareidentified potential porcine enzootic pneumonia determinants

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