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.
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References
1.
AfsetJE, BevangerL, RomundstadP, BerghK. Association of atypical enteropathogenic Escherichia coli (EPEC) with prolonged diarrhea. J Med Microbiol, 2004; 53:1137–1144.
2.
AfsetJE, BruantG, BrousseauR, HarelJ, AnderssenE, BevangerL, BerghK. Identification of virulence genes linked with diarrhea due to atypical enteropathogenic Escherichia coli by DNA microarray analysis and PCR. J Clin Microbiol, 2006; 44:3703–3711.
3.
AltalhiAD, HassanSA. Bacterial quality of raw milk investigated by Escherichia coli and isolates analysis for specific virulence-gene markers. Food Control, 2009; 20:913–917.
4.
AraujoVS, PagliaresVA, QueirozMLP, Freitas-AlmeidaAC. Occurrence of Staphylococcus and enteropathogens in soft cheese commercialized in the city of Rio de Janeiro, Brazil. J Appl Microbiol, 2002; 92:1172–1177.
5.
ArgudínMA, VanderhaeghenW, VandendriesscheS, VandecandelaereI, AndréFX, DenisO, ButayeP. Antimicrobial resistance and population structure of Staphylococcus epidermidis recovered from animals and humans. Vet Microbiol, 2015; 178:105–113.
6.
CarattoliA. Plasmids and the spread of resistance. Int J Med Microbiol, 2013; 303:298–304.
7.
CarvalhoJDG, ViottoWH, KuayeAY. The quality of Minas Frescal cheese produced by different technological processes. Food Control, 2007; 18:262–267.
8.
ChambersHF, DeLeoFR. Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol, 2003; 7:629–641.
9.
ClermontO, BonacorsiS, BingenE. Rapid and simple determination of the Escherichia coli phylogenetic group. Appl Environ Microbiol, 2000; 66:4555–4558.
10.
Clinical and Laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing; 21st Informational Supplement. CLSI document M100-S21. Wayne, PA: CLSI, 2016.
11.
CostaJCM, EspeschitIDF, PieriFA, BenjaminLA, MoreiraMAS. Increase in biofilm formation by Escherichia coli under conditions that mimic the mastitic mammary gland. Ciênc Rural, 2014; 4:666–671.
12.
CraviotoA, GrossRJ, ScotlandSM, RoweB. An adhesive factor found in strains of Escherichia coli belonging to the traditional infantile enteropathogenic serotypes. Curr Microbiol, 1979; 3:95–99.
CunhaCR, ViottoWH, ViottoLA. Use of low concentration factor ultrafiltration retentates in reduced fat “Minas Frescal” cheese manufacture: Effect on composition, proteolysis, viscoelastic properties and sensory acceptance. Int Dairy J, 2006; 16:215–224.
15.
DiasCMA, Sant'AnaAS, CruzAG, FariaJDAF, OliveiraCAF, BonaE. On the implementation of good manufacturing practices in a small processing unity of mozzarella cheese in Brazil. Food Control, 2012; 24:199–205.
16.
FerreiraMA, BernardoLG, NevesLS, CamposMRH, Lamaro-CardosoJ, AndréMCP. Virulence profile and genetic variability of Staphylococcus aureus isolated from artisanal cheese. J Dairy Sci, 2016; 99:8589–8597.
17.
Guzman-HernandezR, Contreras-RodriguezA, Hernandez-VelezR, Perez-MartinezI, Lopez-MerinoA, ZaidiMB, Estrada-GarciaT. Mexican unpasteurised fresh cheeses are contaminated with Salmonella spp., non-O157 Shiga toxin producing Escherichia coli and potential uropathogenic E. coli strains: A public health risk. Int J Food Microbiol, 2016; 237:10–16.
18.
HernandesRT, EliasWP, VieiraMAM, GomesTAT. An overview of atypical enteropathogenic Escherichia coli. FEMS Microbiol Lett, 2009; 297:137–149.
19.
JamaliH, PaydarM, RadmehrB, IsmailS, DadrasniaA. Prevalence and antimicrobial resistance of Staphylococcus aureus isolated from raw milk and dairy products. Food Control, 2015; 54:383–388.
20.
KadariyaJ, SmithTC, ThapaliyaD. Staphylococcus aureus and staphylococcal food-borne disease: An ongoing challenge in public health. BioMed Res Int, 2014; 2014:827965.
LemaîtreC, Mahjoub-MessaiF, DupontD, CaroV, DiancourtL, BingenE, BidetP, BonacorsiS. A conserved virulence plasmidic region contributes to the virulence of the multiresistant Escherichia coli meningitis strain S286 belonging to phylogenetic group C. PLoS One, 2013; 8:74–423.
23.
LevineMM, NataroJP, KarchH, BaldiniMM, KaperJB, BlackRE. The diarrheal response of humans to some classic serotypes of enteropathogenic Escherichia coli is dependent on a plasmid encoding an enteroadhesiveness factor. J Infect Dis, 1985; 3:550–559.
24.
MarozziS, De SantisP, LovariS, CondoleoR, BileiS, MarcianòR, MezherZ. Prevalence and molecular characterization of Shiga toxin-producing Escherichia coli in raw milk cheeses from Lazio region, Italy. Ital J Food Sci, 2016; 5:4566.
25.
MoraesPM, ViçosaGN, YamaziAK, OrtolaniMB, NeroLA. Foodborne pathogens and microbiological characteristics of raw milk soft cheese produced and on retail sale in Brazil. Foodborne Pathog Dis, 2009; 6:245–249.
26.
NagyB, SzmolkaA, SmoleMozinaS, KovacJ, StraussA, SchlagerS, BeutlichJ, AppelB, LusickyM, AprikianP, PasztiJ, TothI, KuglerR, WagnerM. Virulence and antimicrobial resistance determinants of verotoxigenic Escherichia coli (VTEC) and of multidrug-resistant E. coli from foods of animal origin illegally imported to the EU by fligh passengers. Int J Food Microbiol, 2015; 209:52–59.
27.
NataroJP, KaperJB, Robins-BrowneR, PradoV, VialP, LevineMM. Patterns of adherence of diarrheagenic Escherichia coli to HEp-2 cells. Pediatr Infect Dis J, 1987; 6:829–831.
28.
NavarroA, EslavaC, PereaLM, InzunzaA, DelgadoG, Morales-EspinosaR, CraviotoA. New enterovirulent Escherichia coli serogroup 64474 showing antigenic and genotypic relationships to Shigella boydii 16. J Med Microbiol, 2010; 4:453–461.
29.
NobiliG, FranconieriI, BasanisiMG, La BellaG, TozzoliR, CaprioliA, La SalandraG. Short communication: Isolation of Shiga toxin-producing Escherichia coli in raw milk and mozzarella cheese in southern Italy. J Dairy Sci, 2016; 99:7877–7880.
30.
OkuraMH, MarinJM. Survey of Minas frescal cheese from Southwest Minas Gerais for virulence factors and antimicrobial resistance in Escherichia coli isolates. Ciênc Rural (Impress), 2014; 44:1506–1511.
31.
OmbarakRA, HinenoyaA, AwasthiSP, IguchiA, ShimaA, ElbagoryARM, YamasakiS. Prevalence and pathogenic potential of Escherichia coli isolates from raw milk and raw milk cheese in Egypt. Int J Food Microbiol, 2016; 221:69–76.
32.
OrskovF, OrskovI. Serotyping of Escherichia coli. Meth Microbiol, 1984; 14:43–112.
33.
OshoaTJ, ContrerasCA. Enteropathogenic E. coli (EPEC) infection in children. Curr Opin Infect Dis, 2011; 24(5):478–483.
34.
PanetoBR, Schocken-IturrinoRP, MacedoC, SantoE, MarinJM. Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil. Arq Bras Med Vet Zootec, 2007; 59:508–512.
35.
PeixotoMM, GresslerLT, SutiliFJ, CostaMM, VargasAC. Ação dos desinfetantes sobre a adesão e biofilme consolidado de Staphylococcus spp. Pesq Vet Bras, 2015; 2:105–109.
36.
Puño-SarmientoJ, GazalLE, MedeirosLP, NishioEK, KobayashiRK, NakazatoG. Identification of diarrheagenic Escherichia coli strains from avian organic fertilizers. Int J Environ Res Public Health, 2014; 11:8924–8939.
37.
RapiniLS, CerqueiraMMOP, CarmoLS, VerasJF, SouzaMR. Presença de Staphylococcus spp. produtores de enterotoxinas e da toxina da síndrome do choque tóxico em manipuladores de queijo de cabra. Arq Bras Med Vet Zootec, 2005; 57:825–829.
38.
RibeiroLF, BarbosaMMC, PintoFDR, MalutaRP, OliveiraMC, de SouzaV, FairbrotherJM. Antimicrobial resistance and virulence factors of Escherichia coli in cheese made from unpasteurized milk in three cities in Brazil. Foodborne Pathog Dis, 2016; 13:469–476.
39.
ScaletskyIC, PedrosoMZ, OlivaCA, CarvalhoRL, MoraisMB, Fagundes-NetoU. A localized adherence-like pattern as a second pattern of adherence of classic enteropathogenic Escherichia coli to HEp-2 cells that is associated with infantile diarrhea. Infect Immun, 1999; 67:3410–3415.
40.
ScheutzF, CheastyT, WoodwardD and SmithH R. Designation of O174 and O175 to temporary O groups OX3 and OX7, and six new E. coli O groups that include verocytotoxin-producing E. coli (VTEC): O176, O177, O178, O179, O180 and O181. APMIS, 2004; 112:569–584.
41.
SchmidtH, KnopC, FrankeS, AleksicS, HeesemannJ, KarchH. Development of PCR for screening of enteroaggregative Escherichia coli. J Clin Microbiol, 1995; 33:701–705.
42.
SzmolkaA, AnjumMF, La RagioneRM, KaszanyitzkyEJ, NagyB. Microarray based comparative genotyping of gentamicin resistant Escherichia coli strains from food animals and humans. Vet Microbiol, 2012; 156:110–118.
WakimotoN, NishiJ, SheikhJ, NataroJP, SarantuyaJ, IwashitaM, ManagoK, TokudaK, YoshinagaM, KawanoY. Quantitative biofilm assay using a microtiter plate to screen for enteroaggregative Escherichia coli. Am J Trop Med Hyg, 2004; 71:687–690.
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