Identification and detection of mutations potentially associated with decreased susceptibility to macrolides and lincomycin in Mycoplasma anserisalpingitidis isolates

Grózner, Dénes; Bekő, Katinka; Kovács, Áron Botond; Mitter, Alexa; Hrivnák, Veronika; Sawicka, Anna; Tomczyk, Grzegorz; Bányai, Krisztiàn; Jánosi, Szilárd; Kreizinger, Zsuzsa; Gyuranecz, Miklós

doi:10.1016/j.vetmic.2022.109362

Cited by 3 publications

(11 citation statements)

References 24 publications

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“…Increased resistance of biofilms to antibiotics has been described previously in many studies (Costerton et al, 1999;Reid, 1999;Mah and O'Toole, 2001;Stewart, 2002;Sharma et al, 2019;Yan and Bassler, 2019;Daubenspeck et al, 2020). However, no correlation was shown in this study between biofilm-forming ability and the minimal inhibitory concentration values determined previously (Grózner et al, 2016(Grózner et al, , 2022 in the 32 examined M. anserisalpingitidis strains. In case of other bacteria, the nutrient constitution of the growth media was described to affect inhibitory antibiotic concentrations of sessile bacteria (Chen et al, 2020).…”

Section: Discussioncontrasting

confidence: 44%

“…This phenomenon may have contributed to the observed lack of correlation between biofilm-forming ability and antibiotic susceptibility in M. anserisalpingitidis strains, although propagation and examinations were carried out in the nutrient-rich medium which is also used for isolation from clinical samples. Nevertheless, MIC values are primarily affected by resistance-associated genes and mutations, and other resistance mechanisms, such as extracellular vesicles or efflux pumps may also play a role (Medvedeva et al, 2014;Antunes et al, 2015;Grózner et al, 2022). Antibiotic susceptibility of the five tested biofilm-forming M. anserisalpingitidis strains was not clearly affected by the experimental prevention of biofilm formation, in the same way as in the study of McAuliffe et al (2006) with ruminantpathogenic Mycoplasma species.…”

Section: Discussionmentioning

confidence: 72%

“…No correlation was observed between biofilm-forming ability and the MIC values determined previously (Grózner et al, 2016(Grózner et al, , 2022 in the 32 examined M. anserisalpingitidis strains, as high MIC values of each antibiotic were determined against strains lacking this ability as well (Table 3). Accordingly, the antibiotic susceptibility profiles of B and P cultures of the five tested M. anserisalpingitidis strains were highly similar (Table 4), and these strains were mostly inhibited by a concentration close to the previously determined MIC value (no more than one dilution step difference).…”

Section: Effect Of Biofilm Formation On Antibiotic Susceptibilitymentioning

confidence: 91%

“…At first, possible correlations between biofilm-forming ability and the initial or final minimal inhibitory concentration (MICi or MICf) values were analysed. MIC values against the 32 examined M. anserisalpingitidis strains were previously determined (Grózner et al, 2016(Grózner et al, , 2022. The MICi of each strain was defined as the lowest concentration of the antibiotic that completely inhibited the growth at the time of colour change in the growth control (broth culture without antibiotics), while MICf was evaluated at the end of the incubation period (14 days).…”

Section: Investigation Of the Effect Of Biofilm Formation On Antibiot...mentioning

confidence: 99%

“…All products originated from VETRANAL (Sigma-Aldrich Inc., St. Louis, USA) except for tylvalosin (Aivlosin), which was purchased from ECO Animal Health Ltd. (London, UK). The tested concentrations were selected based on the results of our previous studies: 0.078-10 μg mL À1 for enrofloxacin, doxycycline and tiamulin, and 0.125-64 μg mL À1 for the other antibiotics (Grózner et al, 2016(Grózner et al, , 2022. Initial MIC values of each strain were determined by broth microdilution method (Hannan, 2000) with a small modification (using Eppendorf tubes instead of microtitre plates).…”

Section: Investigation Of the Effect Of Biofilm Formation On Antibiot...mentioning

confidence: 99%

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Biofilm formation and its impact on environmental survival and antibiotic resistance of Mycoplasma anserisalpingitidis strains

Bekő

Nagy

Grózner

et al. 2022

AVet

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Several Mycoplasma species can form biofilm, facilitating their survival in the environment, and shielding them from therapeutic agents. The aim of this study was to examine the biofilm-forming ability and its potential effects on environmental survival and antibiotic resistance in Mycoplasma anserisalpingitidis, the clinically and economically most important waterfowl Mycoplasma species. The biofilm-forming ability of 32 M. anserisalpingitidis strains was examined by crystal violet assay. Biofilms and planktonic cultures of the selected strains were exposed to a temperature of 50 °C (20 and 30 min), to desiccation at room temperature (16 and 24 h), or to various concentrations of eight different antibiotics. Crystal violet staining revealed great diversity in the biofilm-forming ability of the 32 tested M. anserisalpingitidis strains, with positive staining in more than half of them. Biofilms were found to be more resistant to heat and desiccation than planktonic cultures, while no correlation was shown between biofilm formation and antibiotic susceptibility. Our results indicate that M. anserisalpingitidis biofilms may contribute to the persistence of the organisms in the environment, which should be taken into account for proper management. Antibiotic susceptibility was not affected by biofilm formation; however, it is important to note that correlations were examined only in vitro.

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Section: Discussioncontrasting

confidence: 44%

Section: Discussionmentioning

confidence: 72%

Section: Effect Of Biofilm Formation On Antibiotic Susceptibilitymentioning

confidence: 91%

Section: Investigation Of the Effect Of Biofilm Formation On Antibiot...mentioning

confidence: 99%

Section: Investigation Of the Effect Of Biofilm Formation On Antibiot...mentioning

confidence: 99%

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Biofilm formation and its impact on environmental survival and antibiotic resistance of Mycoplasma anserisalpingitidis strains

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et al. 2022

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Isolation and characterization of multidrug resistant Gallibacterium anatis biovar haemolytica strains from Polish geese and hens

Karwańska,

Wieliczko,

Bojesen

et al. 2023

Vet Res

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Gallibacterium anatis biovar haemolytica is a bacterium that is frequently associated with infections of the reproductive tract and respiratory system in poultry. To assess the current prevalence and resistance profile of these bacteria in Poland, we collected and investigated 63 strains of Gallibacterium from diseased domestic poultry flocks including geese, laying hens, breeding hens and an ornamental hen. Detailed characterization of the isolates included the analysis of phenotypic antimicrobial resistance profiles and biofilm formation ability. Furthermore, the genetic background of 40 selected isolates regarding the presence of virulence and antimicrobial resistance genes and mobile genetic elements was determined. All investigated isolates were multidrug resistant, most prominently to β-lactams, fluoroquinolones, sulfonamides and macrolides. A total of 48 different resistance profiles were detected. Of all isolates, 50.8% formed a strong biofilm, where strains isolated from geese appeared to be better at biofilm formation than strains isolated from laying and breeding hens. Single-nucleotide polymorphism genotyping revealed that G. anatis bv. haemolytica strains are restricted in host and geographical distribution, and the geese isolates showed greater phylogenetic similarity. Whole genome sequencing enabled identification of 25 different antimicrobial resistance determinants. The most common resistance genes were tetB, blaROB-1, and blaTEM-1 which may be located on mobile genetic elements. All isolates possessed the toxin gene gtxA, and the fimbrial gene flfA was identified in 95% of strains. Our results indicated that all G. anatis bv. haemolytica isolates showed multidrug resistant phenotypes. Strains isolated from geese were characterized by the highest percentage of isolates resistant to selected antimicrobials, probably reflecting host-related adaptations.

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Phenotypic and genetic insights into efflux pump mechanism in Mycoplasma anserisalpingitidis

et al. 2023

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IntroductionMycoplasma anserisalpingitidis is one of the most important waterfowl-pathogenic mycoplasmas. Due to inadequate antibiotic treatment, many strains with high minimal inhibitory concentration (MIC) values for multiple drugs have been isolated lately. Decreased antibiotic susceptibility in several Mycoplasma species are known to be associated with mutations in topoisomerase and ribosomal genes, but other strategies such as active efflux pump mechanisms were also described. The scope of this study was the phenotypic and genetic characterization of the active efflux mechanism in M. anserisalpingitidisMethodsWe measured the MIC values in the presence and absence of different efflux pump inhibitors (EPIs), such as carbonyl cyanide m-chlorophenylhydrazine (CCCP), orthovanadate (OV), and reserpine (RSP). Moreover, bioinformatic tools were utilized to detect putative regulatory sequences of membrane transport proteins coding genes, while comparative genome analysis was performed to reveal potential markers of antibiotic resistance.ResultsOut of the three examined EPIs, CCCP decreased the MICs at least two-fold below the original MICs (in 23 cases out of 36 strains). In the presence of OV or RSP, MIC value differences could be seen only if modified dilution series (10% decrease steps were used instead of two-fold dilutions) were applied (in 24/36 cases with OV and 9/36 with RSP). During comparative genome analysis, non-synonymous single nucleotide polymorphisms (nsSNPs) were identified in genes encoding ABC membrane transport proteins, which were displayed in higher percentages in M. anserisalpingitidis strains with increased MICs. In terms of other genes, a nsSNP was identified in DNA gyrase subunit A (gyrA) gene which can be related to decreased susceptibility to enrofloxacin. The present study is the first to highlight the importance of efflux pump mechanisms in M. anserisalpingitidis.DiscussionConsidering the observed effects of the EPI CCCP against this bacterium, it can be assumed, that the use of EPIs would increase the efficiency of targeted antibiotic therapy in the future control of this pathogen. However, further research is required to obtain a more comprehensive understanding of efflux pump mechanism in this bacterium.

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Identification and detection of mutations potentially associated with decreased susceptibility to macrolides and lincomycin in Mycoplasma anserisalpingitidis isolates

Cited by 3 publications

References 24 publications

Biofilm formation and its impact on environmental survival and antibiotic resistance of Mycoplasma anserisalpingitidis strains

Biofilm formation and its impact on environmental survival and antibiotic resistance of Mycoplasma anserisalpingitidis strains

Isolation and characterization of multidrug resistant Gallibacterium anatis biovar haemolytica strains from Polish geese and hens

Phenotypic and genetic insights into efflux pump mechanism in Mycoplasma anserisalpingitidis

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