Endophytic Lifestyle of Global Clones of Extended-Spectrum beta-Lactamase-Producing Priority Pathogens in Fresh Vegetables: a Trojan Horse Strategy Favoring Human Colonization?

Autor(es)

Lopes, Ralf ^[1] ; Fuentes-Castillo, Danny ^{[2, 3]} ; Fontana, Herrison ^{[4, 3]} ; Rodrigues, Larissa ^{[4, 3]} ; Dantas, Karine ^[1] ; Cerdeira, Louise ^[5] ; Henriques, Isabel ^{[6, 7]} ; Lincopan, Nilton ^{[1, 4, 3]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Biomed Sci, Dept Microbiol, Sao Paulo - Brazil

^[2] Univ Sao Paulo, Sch Vet Med & Anim Sci, Dept Pathol, Sao Paulo - Brazil

^[3] One Hlth Brazilian Resistance Project OneBR, Sao Paulo - Brazil

^[4] Univ Sao Paulo, Sch Pharm, Dept Clin Anal, Sao Paulo - Brazil

^[5] Monash Univ, Cent Clin Sch, Dept Infect Dis, Melbourne, Vic - Australia

^[6] Univ Coimbra, Fac Sci & Technol, Dept Life Sci, Coimbra - Portugal

^[7] Univ Aveiro, Ctr Environm & Marine Studies CESAM, Aveiro - Portugal

• Tipo de documento: Artigo Científico
• Fonte: MSYSTEMS; v. 6, n. 1 JAN-FEB 2021.
• Citações Web of Science: 1

Resumo

The global spread of antibiotic-resistant bacteria and their resistance genes is a critical issue that is no longer restricted to hospital settings, but also represents a growing problem involving environmental and food safety. In this study, we have performed a microbiological and genomic investigation of critical priority pathogens resistant to broad-spectrum cephalosporins and showing endophytic lifestyles in fresh vegetables sold in a country with high endemicity of extended-spectrum beta-lactamases (ESBLs). We report the isolation of international high-risk clones of CTX-M-15-producing Escherichia coli, belonging to clonal complexes CC38 and CC648, and Klebsiella pneumoniae of complex CC307 from macerated tissue of surface-sterilized leaves of spinach, cabbage, arugula, and lettuce. Regardless of species, all ESBL-positive isolates were able to endophytically colonize common bean (Phaseolus vulgaris) seedlings, showed resistance to acid pH, and had a multidrug-resistant (MDR) profile to clinically relevant antibiotics (i.e., broad-spectrum cephalosporins, aminoglycosides, and fluoroquinolones). Genomic analysis of CTX-M-producing endophytic Enrerobacrerales revealed a wide resistome (antibiotics, biocides, disinfectants, and pesticides) and virulome, and genes for endophytic fitness and for withstanding acidic conditions. Transferable IncFIB and IncHI2A plasmids carried bla(CTX-M-15) genes and, additionally, an IncFIB plasmid (named pKP301cro) also harbored genes encoding resistance to heavy metals. These data support the hypothesis that fresh vegetables marketed for consumption can act as a figurative Trojan horse for the hidden spread of international clones of critical WHO priority pathogens producing ESBLs, and/or their resistance genes, to humans and other animals, which is a critical issue within a food safety and broader public and environmental health perspective. IMPORTANCE Extended-spectrum beta-lactamases (ESBL)-producing Enterobacrerales are a leading cause of human and animal infections, being classified as critical priority pathogens by the World Health Organization. Epidemiological studies have shown that spread of ESBL-producing bacteria is not a problem restricted to hospitals, but also represents a growing problem involving environmental and food safety. In this regard, CTX-M-type beta-lactamases have become the most widely distributed and clinically relevant ESBLs worldwide. Here, we have investigated the occurrence and genomic features of ESBL-producing Enterobacterales in surface-sterilized fresh vegetables. We have uncovered that international high-risk clones of CTX-M-15-producing Escherichia coil and Klebsiella pneumoniae harboring a wide resistome and virulome, carry additional genes for endophytic fitness and resistance to acidic conditions. Furthermore, we have demonstrated that these CTX-M-15-positive isolates are able to endophytically colonize plant tissues. Therefore, we believe that fresh vegetables can act as a figurative Trojan horse for the hidden spread of critical priority pathogens exhibiting endophytic lifestyles. (AU)

• Processo FAPESP: 16/08593-9 - Pan-resistoma de Klebsiella pneumoniae e Escherichia coli produtoras de beta-lactamases (KPC-2, CTX-M-8, CTX-M-15) endêmicas no Brasil
• Beneficiário: Nilton Erbet Lincopan Huenuman
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/21325-0 - Análise comparativa do resistoma de Klebsiella pneumoniae XDR (NDM-1/KPC-2) pertencente ao complexo clonal (CC) endêmico de alto risco CC258
• Beneficiário: Louise Teixeira Cerdeira
• Modalidade de apoio: Bolsas no Brasil - Doutorado