RESUMO
Cow raw milk cheese is widely eaten in Brazil. These products may be contaminated with pathogenic bacteria. In this work, we investigated the presence of Escherichia coli in raw milk cheese from different States in Brazil. From 147 "Minas" cheese samples, 28 cheeses were positive for E. coli. Among 39 E. coli isolates of the cheeses, one was positive for eae and negative for bpfA and efa1/lifA using PCR, and so was classified as atypical Enteropathogenic E. coli (aEPEC). Two other isolates were positive for extraintestinal pathogenic E. coli (ExPEC) genes. The aEPEC isolate belongs to serogroup O127 and was classified in A phylogenetic group, and ExPEC isolates were found in O73:H12 (EC-2 strain) and O64474:H8 (EC-9 strain) serotype. This ExPEC belongs to A and C phylogenetic group, respectively. Most of E. coli strains belonged to Clermont phylogenetic groups A (28.2%), C, and E (23.1%). Six strains (15.4%) of E. coli were positive for group B1 and two (5.1%) for B2. E. coli isolates presented an aggregative (46.0%) and diffuse (12.6%) adherence pattern to HeLa cells, and the other isolates did not show adhesion (41.4%). Four E. coli isolates (10.3%) were shown to produce moderate biofilm. The antimicrobial resistance rate was tetracycline (25.6%), followed by ampicillin (17.9%), cefoxitin (7.7%), nalidixic acid (5.1%), and amoxicillin-clavulanic acid (2.6%). One strain was resistant to three antimicrobials (tetracycline, ampicillin, and nalidixic acid). The presence of these microorganisms, the O127 strain, and a new serogroup in Brazil is a potential risk for public health.
Assuntos
Queijo/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Escherichia coli Enteropatogênica/genética , Leite/microbiologia , Combinação Amoxicilina e Clavulanato de Potássio/farmacologia , Ampicilina/farmacologia , Animais , Antibacterianos/farmacologia , Aderência Bacteriana , Brasil , Cefoxitina/farmacologia , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Escherichia coli Enteropatogênica/efeitos dos fármacos , Escherichia coli Enteropatogênica/isolamento & purificação , Contaminação de Alimentos , Microbiologia de Alimentos , Células HeLa , Humanos , Testes de Sensibilidade Microbiana , Ácido Nalidíxico/farmacologia , Pasteurização , Filogenia , Sorogrupo , Sorotipagem , Tetraciclina/farmacologiaRESUMO
Poultry litter is commonly used as fertilizer in agriculture. However, this poultry litter must be processed prior to use, since poultry have a large number of pathogenic microorganisms. The aims of this study were to isolate and genotypically and phenotypically characterize Escherichia coli from avian organic fertilizer. Sixty-four E. coli isolates were identified from avian organic fertilizer and characterized for ExPEC virulence factors, pathogenicity islands, phylogenetic groups, antimicrobial resistance, biofilm formation, and adhesion to HEp-2 cells. Sixty-three isolates (98.4%) showed at least one virulence gene (fimH, ecpA, sitA, traT, iutA, iroN, hlyF, ompT and iss). The predominant phylogenetic groups were groups A (59.3%) and B1 (34.3%). The pathogenicity island CFT073II (51.5%) was the most prevalent among the isolates tested. Thirty-two isolates (50%) were resistant to at least one antimicrobial agent. Approximately 90% of isolates adhered to HEp-2 cells, and the predominant pattern was aggregative adherence (74.1%). In the biofilm assay, it was observed that 75% of isolates did not produce biofilm. These results lead us to conclude that some E. coli isolates from avian organic fertilizer could be pathogenic for humans.
Assuntos
Proteínas de Escherichia coli/genética , Escherichia coli/genética , Fertilizantes/análise , Ilhas Genômicas/genética , Aves Domésticas , Fatores de Virulência/genética , Criação de Animais Domésticos , Animais , Antibacterianos/farmacologia , Brasil , Farmacorresistência Bacteriana , Escherichia coli/efeitos dos fármacos , Proteínas de Escherichia coli/metabolismo , Virulência/genética , Fatores de Virulência/metabolismoRESUMO
The Brazilian poultry industry generates large amounts of organic waste, such as chicken litter, which is often used in agriculture. Among the bacteria present in organic fertilizer are members of the Enterobacteriaceae family. The objective of this study was to detect the presence of diarrheagenic Escherichia coli (DEC) strains in avian organic fertilizer, and assess the potential damage they can cause in humans due to antimicrobial resistance. The presence of DEC pathotypes and phylogenetic groups were detected by multiplex-PCR. Phenotypic assays, such as tests for adhesion, cytotoxicity activity, biofilm formation and especially antimicrobial susceptibility, were performed. Fifteen DEC strains from 64 E. coli were isolated. Among these, four strains were classified as enteropathogenic (EPEC; 6.2%), three strains as Shiga toxin-producing (STEC; 4.7%), 10 strains as enteroaggregative (EAEC; 12.5%), but two of these harbored the eaeA gene too. The low number of isolated strains was most likely due to the composting process, which reduces the number of microorganisms. These strains were able to adhere to HEp-2 and HeLa cells and produce Shiga-toxins and biofilms; in addition, some of the strains showed antimicrobial resistance, which indicates a risk of the transfer of resistance genes to human E. coli. These results showed that DEC strains isolated from avian organic fertilizers can cause human infections.