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Master's Dissertation
DOI
Document
Author
Full name
Maria Elena Espinoza Muñoz
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2019
Supervisor
Committee
Lincopan, Nilton (President)
Garcia, Doroti de Oliveira
Sampaio, Jorge Luiz Mello
Silva, Rodrigo Cayô da
Title in Portuguese
Resistência aos compostos de amônio quaternário (QACs) de uso doméstico e hospitalar em patógenos prioritários multirresistentes
Keywords in Portuguese
CIMs
Compostos de amônio quaternário
Genes QAC
Patógenos prioritários
Resistência
Abstract in Portuguese
Compostos de amônio quaternário (QACs) têm sido amplamente utilizados como desinfetantes e antissépticos, sendo essenciais na prevenção e controle de infecções bacterianas na medicina humana e veterinária. Embora patógenos prioritários multirresistentes têm sido muito bem caracterizados quanto ao perfil de suscetibilidade e contexto genético da resistência aos antibióticos, dados de resistência aos QACs são limitados. Assim, o objetivo do presente estudo foi avaliar a atividade in vitro dos QACs de uso doméstico e hospitalar [cloreto de benzalcônio (BAC), cloreto de cetilpiridinio (CPC) e brometo de cetiltrimetilamônio (CTAB)], contra patógenos prioritários multirresistentes, identificando os principais genes de resistência associados. Foram estudadas 100 cepas multirresistentes previamente sequenciados usando as plataformas Illumina MiSeq e NextSeq representativas de diferentes hospedeiros (humanos e animais) e fontes (ambientes e alimentos). As cepas foram identificadas como Klebsiella pneumoniae (n= 24), Escherichia coli (n= 30); Pseudomonas aeruginosa (n= 10), Enterobacter spp, (n= 8), Acinetobacter baumannii (n= 11) e Salmonella spp. (n= 17). Genes de resistência aos QACs foram identificados in silico através do alinhamento dos contigs obtidos de cada cepa sequenciada com genes de referência obtidos do GenBank, utilizando o programa Geneious versão 8 (Biomatters Ltd). A identidade de cada gene foi analisada utilizando o programa BLASTx, no qual um critério baseado em ≥90% identidade resultou na identificação dos genes mdfA (77%), qacE (44%), qacEΔ1 (43%), sugE(c) (29%), emrE (21%), qacA (19%), sugE(p) (5%), qacF (7%), qacH (7%) e qacL (7%) em 85 cepas; enquanto que 15 cepas não possuíam nenhum gene de resistência aos QACs. A concentração inibitória mínima (CIM) dos QACs para as 100 cepas foi determinada pelo método de microdiluição em caldo. Os resultados sugeriram que a resistência em patógenos prioritários circulando na interface humano-ambiente-animal não é restrita aos antibióticos, uma vez que a elevada ocorrência de genes qacE, qacEΔ1 e mdfA poderia estar associada com uma redução da suscetibilidade para QACs. Consequentemente, a resistência aos QACs poderia também contribuir para a persistência e adaptação destes patógenos nos seres humanos e outros animais, assim como em ambientes impactados antropogenicamente.
Title in English
Resistance to quaternary ammonium compounds (QACs) for domestic and hospital use in multiresistant priority pathogens
Keywords in English
MIC
Priority pathogens
QAC genes
Quaternary ammonium compounds
Resistance
Abstract in English
Quaternary ammonium compounds (QACs) have been widely used as disinfectants and antiseptics, being applied as essential compounds in the prevention and control of bacterial infections in human-and veterinary hospital medicine. Although multiresistant priority pathogens have been well characterized with respect to their susceptibility profile and their genetic context of resistance for antibiotics, studies of resistance to QACs are limited. Thus, the objective of the present study was to evaluate the in vitro activity of QACs [(benzalkonium chloride (BAC), cetylpyridinium chloride (CPC) and cetyltrimethylammonium bromide (CTAB)] for household and hospital use against multiresistant priority pathogens, identifying the main resistance genes associated. A hundred multiresistant isolates (previously sequenced using the Illumina MiSeq and NextSeq platforms), representative of different hosts (humans and animals) and sources (environment and food) were studied. Isolates were identified as Klebsiella pneumoniae (n=24), Escherichia coli (n=30), Pseudomonas aeruginosa (n=10), Enterobacter spp. (n=8), Acinetobacter baumannii (n=11) and Salmonella spp. (n=17). In silico analysis for identification of genes conferring resistance to QACs were performed by aligning the contigs obtained from the strains with reference genes deposited in GenBank, using the Geneious version program (Biomatters Ltd). Similarities were analyzed using the BLASTx online program, considering the alignment criteria based on ≥ 90% identity. The result of these analysis revealed the presence of the following QAC genes: mdfA (77%), qacE (44%), qacEΔ1 (43%), sugE(c) (29%), emrE (21%), qacA (19%), sugE (p) (5%), qacF (7%), qacH (7%) e qacL (7%); while 15 strains showed no resistance genes for QACs. Determination of QACs minimum inhibitory concentration (MIC) for the 100 isolates, by the broth microdilution method. These results suggest that resistance to QACs in priority pathogens, circulating at the human-environment-animal interface, is not restricted to antibiotics, since the high occurrence of genes qacE, qacEΔ1 and mdfA were associated with a reduced susceptibility to QACs. Consequently, resistance to QACs could also contribute to the persistence and adaptation of these pathogens in humans and othes animals, as well as in anthropogenically impacted environments.
 
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Publishing Date
2019-07-11
 
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