Tereza Cristina Rocha Moreira de Oliveira

Comparison of the adhesion ability of different Salmonella enteritidis serotypes to materials used in kitchens

Contamination of kitchen surfaces due to bacteria present in foodstuffs is one of the main causes of foodborne outbreaks. Salmonella infections are an important cause of foodborne disease, and Salmonella Enteritidis is the most common isolate in the past few years. In this study, the adhesion ability of four Salmonella Enteritidis isolates to different materials (polyethylene, polypropylene, and granite) used in kitchens was compared. The results indicated that the two plastic materials were generally less prone to colonization than was the granite. As surface properties of both bacteria and materials are a determinant in the adhesion process, surface hydrophobicity was determined through contact angle measurement, and the roughness of the materials was evaluated through the R(a) and R(z) values by a noncontact laser stylus tracing. The four Salmonella strains showed similar degrees of hydrophilicity, while the materials were hydrophobic, with granite having a very low degree of hydrophobicity (deltaG(lwl) = -4.7 mJ/m2). However, the different extents of adhesion could not be explained in terms of surface hydrophobicity and roughness of the materials tested. The main conclusion to be drawn is that Salmonella adhesion is strongly strain dependent, despite the similar degree of hydrophobicity displayed by all the strains assayed, and this can constitute a factor of virulence among the different serotypes.

Evaluation of fumonisin-aflatoxin co-occurrence in Brazilian corn hybrids by ELISA

The natural co-occurrence of fumonisins and aflatoxins was investigated in freshly harvested corn kernels (150 samples, 62 hybrids), acquired from the Central-Southern (27 samples, 21 hybrids), Central-Western (86 samples, 51 hybrids) and Northern (37 samples, 18 hybrids) regions of the State of Paraná

Adhesion of Salmonella Enteritidis to stainless steel surfaces

Adhesion of microorganisms to food processing surfaces and the problems it causes are a matter of strong concern to the food industry. Contaminated food processing surfaces may act as potential sources of transmission of pathogens in food industry, catering and in the domestic environments. Several studies have shown that adhesion of bacteria to surfaces partly depends upon the nature of the inert surfaces and partly upon the bacterial surface properties. The aim of this study was to compare the adhesion of four different strains of Salmonella Enteritidis to stainless steel 304 (SS 304). The effect of surface hydrophobicity and surface elemental composition on the adhesion process was also analysed. Hydrophobicity was evaluated through contact angle measurements using the sessile drop method. All the strains studied showed positive values of the degree of hydrophobicity (DGlwl) and so can be considered hydrophilic while stainless steel revealed a hydrophobic character. Bacterial cell surface composition was measured using X-ray photoelectron spectroscopy (XPS). The XPS results corroborated the similarity of the values of the degree of hydrophobicity obtained by contact angles. The different Salmonella strains showed similar elemental composition and cell surface physico-chemical properties. Nevertheless, S. Enteritidis MUSC presented higher adhesion ability to SS 304 (p<0.05). It can be concluded that the physico-chemical properties of the strain does not explain the ability of adhesion to stainless steel. Other factors like the production of polysaccharides must be considered.

A comparison between enzyme immunoassay and HPLC for ochratoxin A detection in green, roasted and instant coffee

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for ochratoxin A (OTA) detection in green, roasted and instant coffees was developed using anti-OTA monoclonal antibody. Immunological reagents prepared were OTA-BSA (4.76 mg/mL), anti-OTA.7 MAb (2x103-fold dilution) and HRP-anti IgG (103-fold dilution). The detection limit was 3.73 ng OTA/g and correlation coefficients (r) between this immunoassay and high performance liquid chromatography were 0.98 for green coffee, 0.98 for roasted and 0.86 for instant. OTA levels detected by ic-ELISA were higher than by HPLC, with ELISA/HPLC ratio of 0.66 - 1.46 (green coffee), 0.96 - 1.11 (roasted) and 0.93 - 1.82 (instant). ELISA recoveries for OTA added to coffee (5 - 70 ng/g) were 81.53 % for green coffee, 46.73 % for roasted and 64.35 % for instant, while recoveries by HPLC were 80.54 %, 45.91 % and 55.15 %, respectively. Matrices interferences were minimized by samples dilution before carrying out the ELISA assay. The results indicate that MAb-based ic-ELISA could be a simple, sensitive and specific screening tool for OTA detection, contributing to quality and safety of coffee products.

Ciprofloxacin susceptibility reduction of Salmonella strains isolated from outbreaks

The antimicrobial susceptibility of 212 Salmonella strains isolated from patients and foods was evaluated and 45% were found to be resistant to nalidixic acid. Nalidixic acid resistant strains showed a higher minimal inhibitory concentration for ciprofloxacin than sensitive strains. During the study an increase of strains with reduced susceptibility to ciprofloxacin was also observed.

Contaminação por Salmonella spp. em uma cadeia de produção de ovos de uma integração de postura comercial

Contamination of ISA BROWN layer hens by Salmonella spp. and serotyping of the isolated strains were performed in 30 farms. Salmonella detection was done from feces and cloaca swabs. Eggs produced in the farms contaminated by Salmonella spp. were analyzed by culturing the shell rinsing and egg yolk. Salmonella spp. was isolated in eight (23.0%) farms, but was not detected from eggs. S. Enteritidis, which is the most prevalent sorotype associated to human salmonelosis in Parana State, was not isolated either from birds or eggs analyzed. The sorotypes isolated were Salmonella enterica subsp. enterica (0:3.10) (67.0%), Mbandaka (11.0%), Infantis (11.0%), and Newport (11.0%).

Avaliação do padrão coliformes a 45ºC e comparação da eficiência das técnicas dos tubos múltiplos e Petrifilm EC na detecção de coliformes totais e Escherichia coli em alimentos

Fecal coliforms are defined as coliforms that ferment lactose with gas production within 48 hours at 45oC. Escherichia coli and some strains of Enterobacter and Klebsiella show these same characteristics. However, only the presence of E. coli can indicate fecal contamination of foods, because it is abundantly found in the intestinal tract of all warm-blooded animals, including the humans, and it is not usually found in others niches. The classic denomination of fecal coliforms was changed to coliforms at 45oC in the Resolution no 12/2001 of the National Surveillance Agency of Brazil. The objectives of this study were to analyse the presence of E. coli among the coliforms at 45oC and to compare the methods of Most Probable Number Method (MPN) and Petrifilm EC in their efficiency to detect E. coli in Minas cheese, sausage, fresh vegetables and corn flower samples. Petrifilm EC was more accurate method than MNP method, which presented false negative results or underestimation of E. coli. In conclusion, Petrifilm EC was an efficient and practical method to detect E. coli. Therefore, it may be alternatively used for enumeration of total coliforms and E. coli in foods.

Effect of refrigeration and frozen storage on the Campylobacter jejuni recovery from naturally contaminated broiler carcasses.

Campylobacter jejuni is the most common thermophilic Campylobacter associated with human enteritis in many countries. Broilers and their by-products are the main sources for human enteritis. Refrigeration and freezing are used to control bacterial growth in foods. The effect of these interventions on survival of Campylobacter jejuni is yet not quite understood. This study evaluated the effect of storage temperature on the survival of C. jejuni in chicken meat stored for seven days at 4°C and for 28 days at -20°C. The influence of selective enrichment on recovery of Campylobacter was also evaluated. Thirty fresh chicken meat samples were analyzed and 93.3% was contaminated with termotolerant Campylobacter spp. with average count of 3.08 Log10 CFU/g on direct plating. After refrigeration, 53.3% of the analyzed samples tested positive for Campylobacter and the average count was 1.19 Log10 CFU/g. After storage at -20°C, 36.6% of the samples were positive with a verage count of 0.75 Log10 CFU/g. C. jejuni was detected after enrichment, respectively, in 50% of the fresh, 36.7% of the refrigerated and 33.3% of the frozen meat samples analyzed. No difference was detected for the recovery of C. jejuni from fresh, refrigerated or frozen samples after selective enrichment, showing that this microorganism can survive under the tested storage conditions.

Plasmid-mediated quinolone resistance (PMQR) and mutations in the topoisomerase genes of Salmonella enterica strains from Brazil

The objective of this study was to identify mutations in the Quinolone Resistance Determining sources Regions (QRDR) of the gyrA, gyrB, parC, and parE genes and to determine if any of the qnr variants or the aac(6′)-Ib-cr variant were present in strains of Salmonella spp. isolated in Brazil. A total of 126 Salmonella spp. strains from epidemic (n = 114) and poultry (n = 12) origin were evaluated. One hundred and twelve strains (88.8%) were resistant to nalidixic acid (NAL) and 29 (23.01%) showed a reduced susceptibility to ciprofloxacin (Cip). The mutations identified were substitutions limited to the QRDR of the gyrA gene in the codons for Serine 83, Aspartate 87 and Alanine 131. The sensitivity to NAL seems to be a good phenotypic indication of distinguishing mutated and non-mutated strains in the QRDR, however the double mutation in gyrA did not cause resistance to ciprofloxacin. The qnrA1 and qnrB19 genes were detected, respectively, in one epidemic strain of S. Enteritidis and one strain of S. Corvallis of poultry origin. Despite previous detection of qnr genes in Brazil, this is the first report of qnr gene detection in Salmonella, and also the first detection of qnrB19 gene in this country. The results alert for the continuous monitoring of quinolone resistance determinants in order to minimize the emergence and selection of Salmonella spp. strains showing reduced susceptibility or resistance to quinolones.

Tolerance response of multidrug-resistant Salmonella enterica strains to habituation to Origanum vulgare L. essential oil.

Multidrug-resistant Salmonella enterica isolates from human outbreaks or from poultry origin were investigated for their ability to develop direct-tolerance or cross-tolerance to sodium chloride, potassium chloride, lactic acid, acetic acid, and ciprofloxacin after habituation in subinhibitory amounts ( of the minimum inhibitory concentration – (MIC) and of the minimum inhibitory concentration – MIC) of Origanum vulgare L. essential oil (OVEO) at different time intervals. The habituation of S. enterica to OVEO did not induce direct-tolerance or cross-tolerance in the tested strains, as assessed by the modulation of MIC values. However, cells habituated to OVEO maintained or increased susceptibility to the tested antimicrobials agents, with up to fourfold double dilution decrease from previously determined MIC values. This study reports for the first time the non-inductive effect of OVEO on the acquisition of direct-tolerance or cross-tolerance in multidrug-resistant S. enterica strains to antimicrobial agents that are largely used in food preservation, as well as to CIP, the therapeutic drug of salmonellosis.