Extended Spectrum Beta-Lactamase-Producing Gram-Negative Bacteria Recovered From an Amazonian Lake Near the City of Belém, Brazil

Abstract

Aquatic systems have been described as antibiotic resistance reservoirs, where water may act as a vehicle for the spread of resistant bacteria and resistance genes. We evaluated the occurrence and diversity of third generation cephalosporin-resistant gram-negative bacteria in a lake in the Amazonia region. This water is used for human activities, including consumption after appropriate treatment. Eighteen samples were obtained from six sites in October 2014. Water quality parameters were generally within the legislation limits. Thirty-three bacterial isolates were identified as Escherichia ( n = 7 isolates), Acinetobacter , Enterobacter , and Klebsiella ( n = 5 each), Pseudomonas ( n = 4), Shigella ( n = 3), and Chromobacterium , Citrobacter , Leclercia , Phytobacter (1 isolate each). Twenty nine out of 33 isolates (88%) were resistant to most beta-lactams, except carbapenems, and 88% ( n = 29) were resistant to antibiotics included in at least three different classes. Among the beta-lactamase genes inspected, the bla CTX–M was the most prevalent ( n = 12 positive isolates), followed by bla TEM ( n = 5) and bla SHV ( n = 4). bla CTX–M–15 ( n = 5), bla CTX–M–14 ( n = 1) and bla CTX–M–2 ( n = 1) variants were detected in conserved genomic contexts: bla CTX–M–15 flanked by IS Ecp1 and Orf477; bla CTX–M–14 flanked by IS Ecp1 and IS 903 ; and bla CTX–M–2 associated to an ISCR element. For 4 strains the transfer of bla CTX–M was confirmed by conjugation assays. Compared with the recipient, the transconjugants showed more than 500-fold increases in the MICs of cefotaxime and 16 to 32-fold increases in the MICs of ceftazidime. Two isolates ( Escherichia coli APC43A and Acinetobacter baumannii APC25) were selected for whole genome analysis. APC43A was predicted as a E. coli pathogen of the high-risk clone ST471 and serotype O154:H18. bla CTX–M–15 as well as determinants related to efflux of antibiotics, were noted in APC43A genome. A. baumannii APC25 was susceptible to carbapenems and antibiotic ...

Resistome in Lake Bolonha, Brazilian Amazon: Identification of Genes Related to Resistance to Broad-Spectrum Antibiotics

“…These genes encode resistance to carbapenems, a group of antibiotics commonly used as a last resort for the treatment of serious infections (Tacão et al, 2015). At the P3 site, we also detected the bla CTX−M gene, which encodes ESBLs whose occurrence in the lake Água Preta, adjacent to Lake Bolonha, has also been recently reported (Freitas et al, 2019).…”

“…In addition, culture-dependent methods allow the isolation of target bacteria to study their phenotypic and genotypic characteristics related to antibiotic resistance (McLain et al, 2016). The application of these associated methodologies allows the characterization of the microbiome and resistome, such as in the Brazilian Amazon region, which, despite having the largest hydrographic basin in the world, has rarely been addressed concerning resistance to antibiotics (de Lima et al, 2016;Freitas et al, 2019).…”

Distribution of Beta-Lactamase Producing Gram-Negative Bacterial Isolates in Isabela River of Santo Domingo, Dominican Republic

“…Aliquots of 1, 10, and 50 mL were vacuum-filtered in triplicates through a 0.22 μm pore size nitrocellulose membrane (Simsii, INC) as recommended by Freitas et al ( 2019 ). Bacteria retained in membranes were grown in two culture media: (a) MacConkey agar supplemented with imipenem (IMP) (4 μg/mL); and (b) MacConkey supplemented with cefotaxime (CTX) (8 μg/mL).…”

“…Antibiotic resistance genes (ARGs) are increasingly common in aquatic environments, especially genes of Extended Spectrum β-Lactamases (ESBLs) that are responsible for multi-resistant infections (Chouchani et al, 2013 ). One of the most urgent concerns in the study of ARGs is the ubiquity of ESBLs in urban rivers and lakes (Tacão et al, 2012 ; Freitas et al, 2019 ; Reddy and Dubey, 2019 ). The interaction between anthropogenic effluents (such as the hospital, abattoir, industrial, or residential wastewaters) and environmental waters such as rivers and lakes promotes the formation of resistance genes reservoirs (Marti et al, 2014 ).…”

Genomic insights of Klebsiella pneumoniae isolated from a native Amazonian fish reveal wide resistome against heavy metals, disinfectants, and clinically relevant antibiotics

“…Additionally, some fish species, as the Piraíba ( Brachyplatystoma filamentosum ), the largest catfish of the Amazonian basin, contribute with protein food supply [ 2 ]. Worryingly, recent surveillance studies of antimicrobial resistance in aquatic environments of the Brazilian Amazon have revealed the presence of clinically relevant multidrug-resistant pathogens, including extended-spectrum β-lactamase (ESBL)-producing Enterobacterales [ [3] , [4] , [5] ], which have been recognized as critical-priority pathogens by the World Health Organization (WHO) [ 6 ]. In this study, we report for the first time, the identification and genomic features of a multidrug-resistant (MDR) ESBL-producing Klebsiella pneumoniae strain isolated from a native freshwater catfish commercialized for human consumption in a wet market of the Amazonian region of Brazil, denoting a hidden risk for the human health.…”