Morphological, biochemical, physiological and molecular aspects of the response of Fusobacterium nucleatum exposed to subinhibitory concentrations of antimicrobials

Filho, Job Alves de Souza; Diniz, Cláudio Galuppo; Barbosa, Natália Bento; Freitas, Michele Cristine Ribeiro de; Neves, Mariana Silva Lopes; Mazzei, Rafaella Nogueira da Gama; Gameiro, Jacy; Coelho, Cíntia Marques; Silva, Vânia Lúcia da

doi:10.1016/j.anaerobe.2012.09.007

Cited by 10 publications

(4 citation statements)

References 28 publications

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“…Although some studies have shown that sub-MICs of both PTZ and MET decreased the formation of biofilm and that CLI may increase the same, our results were not able to demonstrate these effects, probably due to the low number of strains tested (OKAMOTO et al, 2002;DE SOUZA FILHO et al, 2012;DE ANDRADE et al, 2016).…”

Section: Discussioncontrasting

confidence: 96%

“…This may not be able to eliminate the microorganism but could interfere with the expression of some pathogenicity factors and may result in the selection of resistant bacteria. These changes in the pathogenicity of B. fragilis, in the presence of sub-MICs of antimicrobials demonstrate their pathogenic potential, being able to provoke different interactions between the host and the bacterium (DE SOUZA FILHO et al, 2012;VELOSO et al, 2013).…”

Section: Discussionmentioning

confidence: 95%

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AVALIAÇÃO DA TOXICIDADE AGUDA E DA CITOTOXICIDADE DOS EXTRATOS ETANÓLICOS DA MACRÓFITA Hydrocotyle bonariensis LAM (APIACEAE)

Nascimento¹,

Guedes²,

Bezerra³

et al. 2020

Pesquisa Científica E Tecnológica Em Microbiologia 3

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Todo o conteúdo deste livro está licenciado sob uma Licença de Atribuição Creative Commons. Atribuição 4.0 Internacional (CC BY 4.0). O conteúdo dos artigos e seus dados em sua forma, correção e confiabilidade são de responsabilidade exclusiva dos autores. Permitido o download da obra e o compartilhamento desde que sejam atribuídos créditos aos autores, mas sem a possibilidade de alterá-la de nenhuma forma ou utilizá-la para fins comerciais.

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Section: Discussioncontrasting

confidence: 96%

Section: Discussionmentioning

confidence: 95%

AVALIAÇÃO DA TOXICIDADE AGUDA E DA CITOTOXICIDADE DOS EXTRATOS ETANÓLICOS DA MACRÓFITA Hydrocotyle bonariensis LAM (APIACEAE)

Nascimento¹,

Guedes²,

Bezerra³

et al. 2020

Pesquisa Científica E Tecnológica Em Microbiologia 3

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“…Add to that, the resident microbiota exposure to subinhibitory concentrations (SIC), which may interfere with microbial ecosystem, may also lead to changes in bacteria-bacteria and bacteria-host relationships related to pleiotropic regulation in bacterial gene expression, as an adaptive response ( Bohnen, 1998 ; Diniz et al, 2004 ). Under SIC of antimicrobial drugs, in vitro studies reported cellular alterations related to the anaerobic bacteria morphology, physiology, and protein expression ( Diniz et al, 2003 ; de Souza Filho et al, 2012 ; Freitas et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Exploratory Investigation of Bacteroides fragilis Transcriptional Response during In vitro Exposure to Subinhibitory Concentration of Metronidazole

et al. 2016

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Bacteroides fragilis, member from commensal gut microbiota, is an important pathogen associated to endogenous infections and metronidazole remains a valuable antibiotic for the treatment of these infections, although bacterial resistance is widely reported. Considering the need of a better understanding on the global mechanisms by which B. fragilis survive upon metronidazole exposure, we performed a RNA-seq transcriptomic approach with validation of gene expression results by qPCR. Bacteria strains were selected after in vitro subcultures with subinhibitory concentration (SIC) of the drug. From a wild type B. fragilis ATCC 43859 four derivative strains were selected: first and fourth subcultures under metronidazole exposure and first and fourth subcultures after drug removal. According to global gene expression analysis, 2,146 protein coding genes were identified, of which a total of 1,618 (77%) were assigned to a Gene Ontology term (GO), indicating that most known cellular functions were taken. Among these 2,146 protein coding genes, 377 were shared among all strains, suggesting that they are critical for B. fragilis survival. In order to identify distinct expression patterns, we also performed a K-means clustering analysis set to 15 groups. This analysis allowed us to detect the major activated or repressed genes encoding for enzymes which act in several metabolic pathways involved in metronidazole response such as drug activation, defense mechanisms against superoxide ions, high expression level of multidrug eﬄux pumps, and DNA repair. The strains collected after metronidazole removal were functionally more similar to those cultured under drug pressure, reinforcing that drug-exposure lead to drastic persistent changes in the B. fragilis gene expression patterns. These results may help to elucidate B. fragilis response during metronidazole exposure, mainly at SIC, contributing with information about bacterial survival strategies under stress conditions in their environment.

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“…In this context, high-throughput sequencing is thought to be useful to evaluate the bacterial transcriptome during an actual infection representing a new generation of genome-wide gene expression measurements based on mRNA sequencing (RNA-seq) (Sȋrbu et al, 2012). These approaches are especially useful when considering resident microbiota as these bacteria can be modulated by host physiology and by xenobiotics such as antimicrobial drugs (Diniz et al, 2003;dos Santos et al, 2010;de Souza Filho et al, 2012;Santos et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Integrity of prokaryotic mRNA isolated from complex samples for in vivo bacterial transcriptome analysis

et al. 2015

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ABSTRACT. Even though several in vitro studies have focused on bacterial biology, the extent of such knowledge is not complete when considering an actual infection. As culture-independent microbiology methods such as high-throughput sequencing became available, important aspects of host-bacterium interactions will be elucidated. Based on microbiological relevance, we considered Bacteroides fragilis in a murine experimental infection as a model system to evaluate the in vivo bacterial transcriptome in host exudates. A disproportionate number of reads belonging to the host genome were retrieved in the first round of pyrosequencing, even after depletion of ribosomal RNA; the average number of reads related to the eukaryotic genome was 71.924-67.7%, whereas prokaryotic reads represented 34.338-32.3% in host exudates. Thus, different treatments were used to improve the prokaryotic RNA yield: i) centrifugation; ii) ultrasonic treatment; and iii) ultrasonic treatment followed by centrifugation. The latter treatment was found to be the most efficient in generating bacterial yields, as it resulted in a higher number of Bacteroides cells. However, the RNA extracted after this treatment was not of sufficient quality to be used in cDNA synthesis. Our results suggest that the methodology routinely used for RNA extraction in transcriptional analysis is not appropriate for in vivo studies in complex samples. Furthermore, the most efficient treatment for generating good bacterial cell yields was not suitable to retrieve high-quality RNA. Therefore, as an alternative methodological approach to enable in vivo studies on host-bacterium interactions, we advise increasing the sequencing depth despite the high costs.

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Morphological, biochemical, physiological and molecular aspects of the response of Fusobacterium nucleatum exposed to subinhibitory concentrations of antimicrobials

Cited by 10 publications

References 28 publications

AVALIAÇÃO DA TOXICIDADE AGUDA E DA CITOTOXICIDADE DOS EXTRATOS ETANÓLICOS DA MACRÓFITA Hydrocotyle bonariensis LAM (APIACEAE)

AVALIAÇÃO DA TOXICIDADE AGUDA E DA CITOTOXICIDADE DOS EXTRATOS ETANÓLICOS DA MACRÓFITA Hydrocotyle bonariensis LAM (APIACEAE)

Exploratory Investigation of Bacteroides fragilis Transcriptional Response during In vitro Exposure to Subinhibitory Concentration of Metronidazole

Integrity of prokaryotic mRNA isolated from complex samples for in vivo bacterial transcriptome analysis

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