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This research analyzed, optimized and modeled the inactivation kinetics of pathogenic bacteria (PB1: Escherichia coli O157:H7 and PB2: Listeria monocytogenes) and determined the microbiological safety of tomato juice processed by UV-LED irradiation and heat treatment. UV-LED processing conditions were optimized using response surface methodology (RSM) and were 90% power intensity, 21 min and 273-275 nm (251 mJ/cm2) with R2 > 0.96. Using the optimal conditions, levels of PB1 and PB2 resulted a log reduction of 2.89 and 2.74 CFU/mL, respectively. The Weibull model was efficient for estimating the log inactivation of PB1 and PB2 (CFU/mL). The kinetic parameter δ showed that 465.2 mJ/cm2 is needed to achieve a 90% log (CFU/mL) reduction in PB1 and 511.3 mJ/cm2 for PB2. With respect to the scale parameter p > 1, there is a descending concave curve. UV-LED-treated tomato juice had an 11.4% lower Listeria monocytogenes count than heat-treated juice on day 28 (4.0 ± 0.82 °C). Therefore, UV-LED technology could be used to inactivate Escherichia coli O157:H7 and Listeria monocytogenes, preserving tomato juice for microbiological safety, but studies are required to further improve the inactivation of these pathogens and analyze other fruit and vegetable juices.
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Introduction. The different pathotypes of Escherichia coli can produce a large number of human diseases. Surveillance is complex since their differentiation is not easy. In particular, the detection of Shiga toxin-producing Escherichia coli (STEC) serotype O157â:âH7 consists of stool culture of a diarrhoeal sample on enriched and/or selective media and identification of presumptive colonies and confirmation, which require a certain level of training and are time-consuming and expensive.Hypothesis. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a quick and easy way to obtain the protein spectrum of a microorganism, identify the genus and species, and detect potential biomarker peaks of certain characteristics.Aim. To verify the usefulness of MALDI-TOF MS to rapidly identify and differentiate STEC O157â:âH7 from other E. coli pathotypes.Methodology. The direct method was employed, and the information obtained using Microflex LT platform-based analysis from 60 clinical isolates (training set) was used to detect differences between the peptide fingerprints of STEC O157â:âH7 and other E. coli strains. The protein profiles detected laid the foundations for the development and evaluation of machine learning predictive models in this study.Results. The detection of potential biomarkers in combination with machine learning predictive models in a new set of 142 samples, called 'test set', achieved 99.3â% (141/142) correct classification, allowing us to distinguish between the isolates of STEC O157â:âH7 and the other E. coli group. Great similarity was also observed with respect to this last group and the Shigella species when applying the potential biomarkers algorithm, allowing differentiation from STEC O157â:âH7Conclusion. Given that STEC O157â:âH7 is the main causal agent of haemolytic uremic syndrome, and based on the performance values obtained in the present study (sensitivity=98.5â% and specificity=100.0â%), the implementation of this technique provides a proof of principle for MALDI-TOF MS and machine learning to identify biomarkers to rapidly screen or confirm STEC O157â:âH7 versus other diarrhoeagenic E. coli in the future.
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Infecciones por Escherichia coli , Escherichia coli O157 , Escherichia coli Shiga-Toxigénica , Humanos , Escherichia coli O157/metabolismo , Serogrupo , Infecciones por Escherichia coli/diagnóstico , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Biomarcadores/metabolismoRESUMEN
Lactic acid bacteria (LAB) selected on the basis of probiotic characteristics were administered to beef feedlot catlle and the effect on body condition/growth and nutritional-metabolic status as well as on E. coli O157:H7 fecal shedding, were investigated. A feeding trials involving 126 steers were used to evaluate the effects of Lactobacillus acidophilus CRL2074, Limosilactobacillus fermentum CRL2085 and Limosilactobacillus mucosae CRL2069 and their combinations (5 different probiotic groups and control) when 107-108 CFU/animal of each probiotic group were in-feed supplemented. Cattle were fed a high energy corn-based diet (16 to 88%) and samples from each animal were taken at 0, 40, 104 and 163 days. In general, animals body condition and sensorium state showed optimal muscle-skeletal development and behavioral adaption to confinement; no nasal/eye discharges and diarrheic feces were observed. The nutritional performance of the steers revealed a steady increase of biometric parameters and weight. Animals supplied with L. mucosae CRL2069 for 104 days reached the maximum mean live weight (343.2 kg), whereas the greatest weight daily gain (1.27 ± 0.16 Kg/day) was obtained when CRL2069 and its combination with L. fermentum CRL2085 (1.26 ± 0.11 kg/day) were administered during the complete fattening cycle. With several exceptions, bovine cattle blood and serum parameters showed values within referential ranges. As a preharvest strategy to reduce Escherichia coli O157:H7 in cattle feces, CRL2085 administered during 40 days decreased pathogen shedding with a reduction of 43% during the feeding period. L. fermentum CRL2085 and L. mucosae CRL2069 show promise for feedlot cattle feeding supplementation to improve metabolic-nutritional status, overall productive performance and to reduce E. coli O157:H7 shedding, thus decreasing contamination chances of meat food products.
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Enfermedades de los Bovinos , Infecciones por Escherichia coli , Probióticos , Bovinos , Animales , Escherichia coli , Alimentación Animal/análisis , Probióticos/farmacología , Suplementos Dietéticos , Heces/microbiología , Enfermedades de los Bovinos/prevención & control , Enfermedades de los Bovinos/microbiología , Recuento de Colonia Microbiana/veterinaria , Infecciones por Escherichia coli/veterinariaRESUMEN
Lactic acid bacteria (LAB) exert antagonistic activities against diverse microorganisms, including pathogens. In this work, we aimed to investigate the ability of LAB strains isolated from food to produce biofilms and to inhibit growth and surface colonization of Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 at 10°C. The ability of 100 isolated LAB to inhibit EHEC O157:H7 NCTC12900 growth was evaluated in agar diffusion assays. Thirty-seven LAB strains showed strong growth inhibitory effect on EHEC. The highest inhibitory activities corresponded to LAB strains belonging to Lactiplantibacillus plantarum, Pediococcus acidilactici and Pediococcus pentosaceus species. Eighteen out of the 37 strains that showed growth inhibitory effects on EHEC also had the ability to form biofilms on polystyrene surfaces at 10°C and 30°C. Pre-established biofilms on polystyrene of four of these LAB strains were able to reduce significantly surface colonization by EHEC at low temperature (10°C). Among these four strains, Lact. plantarum CRL 1075 not only inhibited EHEC but also was able to grow in the presence of the enteric pathogen. Therefore, this strain proved to be a good candidate for further technological studies oriented to its application in food-processing environments to mitigate undesirable surface contaminations of E. coli.
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Antibiosis , Biopelículas/crecimiento & desarrollo , Escherichia coli O157/crecimiento & desarrollo , Lactobacillales/fisiología , Manipulación de Alimentos , Microbiología de Alimentos , Interacciones Microbianas , ProbióticosRESUMEN
Resumen Escherichia coli 0157:H7 es una bacteria patógena reconocida por su capacidad de resistencia a diversos antibióticos; razón por la cual, se generan complicaciones en el tratamiento de infecciones producidas por esta bacteria. El péptido Ib-M1 y el bioconjugado I0NP@Ib-M1 han surgido como una nueva alternativa antimicrobiana contra E. coli 0157:H7. El mecanismo de acción de Ib-Mi e I0NP@Ib-M1 contra esta bacteria aún es desconocido; por lo tanto, el objetivo de esta investigación fue identificar el cambio en el perfil de proteínas de E. coli 0157:H7 luego del tratamiento con Ib-M1 e I0NP@ Ib-M1 como primer paso para determinar su mecanismo de acción. Para esto, se llevó a cabo la obtención de proteínas, posteriormente se realizó una electroforesis bidimensional para finalmente realizar la determinación de la variabilidad de los perfiles proteicos. Una vez obtenidos estos perfiles, se llevó a cabo un análisis de varianza (AN0VA). Se identificaron 72 proteínas expresadas diferencialmente, las cuales pueden relacionarse con el efecto sobre el crecimiento de la bacteria en presencia de Ib-M1 e I0NP@Ib-M. Estas proteínas se encuentran involucradas en procesos de transferencia de grupos acilo (proteína Yhbs), translocación de lipoproteínas (proteína LolA) y transporte de aminoácidos (proteína GpmA), entre otros.
Abstract Escherichia coli 0157: H7 is a pathogenic bacterium which is recognized for the ability to resist multiple antibiotics; accordingly, complications occur in the treatment of infections caused by this bacterium. The Ib-M1 peptide and the I0NP @ Ib-M1 bioconjugate have emerged as a new antimicrobial alternatives against E. coli 0157: H7. The mechanism of action of Ib-M1 and I0NP @ Ib-M1 against this bacterium is still unknown; therefore, the goal of this research was to identify the change in the proteins profile of E. coli 0157: H7 after treatment with Ib-M1 and I0NP @ Ib-M1 as a first step to determine its mechanism of action. For this, the proteins were obtained first, and then a two-dimensional electrophoresis was performed to finally determine the variability of the protein profiles. 0nce the protein profiles were obtained, an analysis of variance (AN0VA) was carried out. 72 differentially expressed proteins were identified, which can be connected to the effect on the bacterium's growth in the presence of Ib-M1 and I0NP @ Ib-M. These proteins are involved in acyl groups transfer processes (Yhbs protein), lipoprotein translocation (LolA protein) and amino acid transport (GpmA protein), among others.
Resumo Escherichia coli O157: H7 é uma bactéria patogênica reconhecida por sua capacidade de resistir a vários antibióticos; razão pela qual, complicações são geradas no tratamento de infecções produzidas por essa bactéria. O peptídeo Ib-M1 livre e imobilizado em nanopartículas magnéticas de óxido de ferro (IONP @ Ib-M1) surgiu como uma nova alternativa antimicrobiana contra E. coli O157: H7 e isolados clínicos desta bactéria. O mecanismo de ação de Ib-M1 e IONP @ Ib-M1 contra E. coli O157: H7 ainda é desconhecido; Portanto, o objetivo desta pesquisa foi identificar a alteração no perfil proteico de E. coli O157: H7 após o tratamento com Ib-M1 e IONP @ Ib-M1 como um primeiro passo para determinar seu mecanismo de ação. Para isso, foi realizada a obtenção das proteínas, posteriormente foi realizada uma eletroforese bidimensional para finalmente determinar a variabilidade dos perfis protéicos. Uma vez obtidos os perfis de proteínas, foi realizada uma análise de variância (ANOVA). Os resultados mostram a identificação de proteínas expressas diferencialmente e que estão envolvidas em processos de transferência de grupos acila (proteína Yhbs), translocação de lipoproteínas (proteína LolA) e transporte de aminoácidos (proteína GpmA), entre outros.
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The bacteriophage UFV-AREG1 was used as a model organism to evaluate the encapsulation via extrusion using different hydrocolloids. Pure alginate [0.75%, 1.0%, 1.5% and 2.0% (m/v)] and mixtures of alginate [0.75% or 1.0% (m/v)] with carrageenan [1.25% (m/v)], chitosan [0.5% (m/v)], or whey protein [1.5% (m/v)] were used to produce bacteriophage-loaded beads. The encapsulating solutions presented flow behavior of non-Newtonian pseudoplastic fluids and the concentration of hydrocolloid did not influence (p > 0.05) the morphology of the beads, except for alginate-chitosan solutions, which presented the higher flow consistency index (K) and the lower flow behavior index (n). The encapsulation efficiency was about 99% and the confocal photomicrography of the encapsulated bacteriophages labeled with fluorescein isothiocyanate showed homogenous distribution of the viral particles within the beads. The phages remained viable in the beads of alginate-whey protein even when submitted to pH 2.5 for 2 h. Beads incubated directly in simulated intestinal fluid (pH 6.8) resulted in a minimal of 50% release of the UFV-AREG1 phages after 5 min, even when previously submitted to the simulated gastric fluid (pH 2.5). Encapsulation enabled phages to remain viable under refrigeration for five months. Encapsulated UFV-AREG1 phages were sensitive to dehydration, suggesting the need for protective agents. In this study, for the first-time bacteriophages were encapsulated in alginate-carrageenan beads, as well as alginate-chitosan as a bead-forming hydrocolloid. In addition, a novel procedure for encapsulating bacteriophages in alginate-whey protein was proposed. The assembled system showed efficiency in the encapsulation of UFV-AREG1 bacteriophages using different hydrocolloids and has potential to be used for the entrapment of a variety of bioactive compounds.
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Alginatos , Bacteriófagos , Preparaciones de Acción Retardada , Ácido Glucurónico , Ácidos Hexurónicos , PolímerosRESUMEN
This study investigated the isolated effect of modified atmosphere packaging (MAP; 50% CO2 and 50% N2) and ultraviolet radiation (UV; 0.30 J/cm2) as well as their combined (MAP/UV) effect on reduction of Salmonella typhimurium and Escherichia coli O157:H7, biogenic amines (BA), and on shelf life of tilapia fillets stored at 4 ± 1 °C for 10 days. UV samples had the highest reduction of S. typhimurium (1.13 log colony forming units/g; CFU/g) and E. coli O157:H7 (0.70 log CFU/g). MAP and MAP/UV reduced the growth of S. typhimurium in 0.50 log CFU/g and did not affect the growth of E. coli O157:H7. UV, MAP, and MAP/UV increased lag phase and/or generation time of all evaluated bacterial groups, decreased pH values, ammonia formation, texture changes, and, in general, the BA formation throughout storage period, and, therefore, UV, MAP, and MAP/UV extended the shelf life for two, three, and at least five days, respectively. MAP/UV, MAP, and UV decreased redness, MAP/UV and MAP increased yellowness and lipid oxidation, while UV did not affect it. MAP/UV demonstrated promising results for shelf life extension; however, different gas ratios in combination with other ultraviolet radiation type C (UV-C) doses should be investigated to reach the highest microbiological safety and maintenance of the overall quality of tilapia fillets.
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Atmósfera , Aminas Biogénicas/análisis , Embalaje de Alimentos , Conservación de Alimentos , Alimentos Marinos/análisis , Tilapia , Rayos Ultravioleta , Animales , Análisis de los AlimentosRESUMEN
The different morphological characteristics of five bacterial pathogen strains were analyzed through transmission electron microscopy for addressing the particular relationship between optical density and colony-forming units for each strain. Generated linear equations will allow a reliable calculation of bacterial concentrations through simple optical density measurements.
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Escherichia coli O157/aislamiento & purificación , Microbiología de Alimentos , Enfermedades Transmitidas por los Alimentos/microbiología , Listeria monocytogenes/aislamiento & purificación , Microscopía Electrónica de Transmisión/métodos , Salmonella typhimurium/aislamiento & purificación , Staphylococcus aureus/aislamiento & purificación , Recuento de Colonia Microbiana/métodos , Manipulación de Alimentos/métodosRESUMEN
The growing demand for fruit and vegetable juice blends, with improved nutritional and sensory attributes, has prompted the industrial adoption of nonthermal processing technologies, including UV light. Limited studies have explored conditions to overcome the well-known limitations of UV when treating liquid foods with a high content of particles that absorb or scatter UV light. This study addressed the effectiveness of the application of UV light, using a commercial processing unit, to inactivate pathogenic Escherichia coli O157:H7, Salmonella enterica (hereafter Salmonella), and Listeria monocytogenes, as well as spoilage microorganisms, in colored and turbid juices and beverages. The inactivation of cocktails of five strains (or serotypes) of E. coli O157:H7, Salmonella, and L. monocytogenes isolated from fruit- and vegetable-derived products linked to outbreaks was determined in seven colored and turbid cold-pressed juices and beverages. Juices and beverages were UV treated at a constant flow rate of 150 L/h through multiple consecutive passes. The inactivation of aerobic mesophilic bacteria, molds and yeasts, and lactic acid bacteria was also assessed at the cumulative dose that guaranteed a 5-log reduction of the most UV-tolerant pathogen for each product. A 5-log reduction of the three pathogens was achieved in all juices and beverages at a maximum cumulative UV dose of 12.0 ± 0.6 mJ/cm2. The dose required to ensure the targeted reduction varied depending on the tested product and the inoculated pathogen. The reduction of aerobic mesophiles, molds and yeasts, and lactic acid bacteria varied from 0.5 to 3.6, from 0.2 to 2.0, and from 0.5 to 3.6 log CFU/mL, respectively. Thus, the proposed treatment represents a suitable processing alternative to ensure the safety and extend the shelf life of colored and turbid cold-pressed juices and beverages.
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Jugos de Frutas y Vegetales , Rayos Ultravioleta , Recuento de Colonia Microbiana , Escherichia coli O157/efectos de la radiación , Microbiología de Alimentos/métodos , Jugos de Frutas y Vegetales/microbiología , Jugos de Frutas y Vegetales/normas , Humanos , Listeria monocytogenes/efectos de la radiación , Viabilidad Microbiana/efectos de la radiaciónRESUMEN
Objective To develop a bioluminescence-labelled bacterial infection model to monitor the colonization and clearance process of Escherichia coli O157:H7 in the lungs of mice following influenza A virus/Puerto Rico/8/34 (H1N1) strain (IAV/PR8) infection. Methods BALB/c mice were administered IAV/PR8 or 0.01 M phosphate-buffered saline (PBS; pH 7.4) intranasally 4 days prior to intranasal administration of 1 × 107 colony-forming units (CFU) of E. coli O157:H7-lux. Whole-body bioluminescent signals were monitored at 10 min, 4 h, 8 h, 12 h, 16 h and 24 h post-bacterial infection. Lung bioluminescent signals and bacterial load (CFU/g) were monitored at 4 h, 8 h, 12 h, 16 h and 24 h post-bacterial infection. Results Prior IAV/PR8 infection of mice resulted in a higher level of bacterial colonization and a lower rate of bacterial clearance from the lungs compared with mice treated with PBS. There were also consistent findings between the bioluminescence imaging and the CFU measurements in terms of identifying bacterial colonization and monitoring the clearance dynamics of E. coli O157:H7-lux in mouse lungs. Conclusion This novel bioluminescence-labelled bacterial infection model rapidly detected bacterial colonization of the lungs and monitored the clearance dynamics of E. coli O157:H7-lux following IAV/PR8 infection.
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Coinfección/microbiología , Coinfección/virología , Infecciones por Escherichia coli/microbiología , Escherichia coli O157/aislamiento & purificación , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Pulmón/microbiología , Pulmón/fisiopatología , Infecciones por Orthomyxoviridae/virología , Animales , Modelos Animales de Enfermedad , Femenino , Mediciones Luminiscentes , Ratones , Ratones Endogámicos BALB CRESUMEN
To clarify the interactions between a common food spoilage yeast and two pathogenic bacteria involved in outbreaks associated with fruit juices, the present paper studies the effect of the interplay of Candida krusei, collected from UF membranes, with Escherichia coli O157:H7 and Salmonella enterica in the overall process of adhesion and colonization of abiotic surfaces. Two different cases were tested: a) co-adhesion by pathogenic bacteria and yeasts, and b) incorporation of bacteria to pre-adhered C. krusei cells. Cultures were made on stainless steel at 25°C using apple juice as culture medium. After 24 h of co-adhesion with C. krusei, both E. coli O157:H7 and S. enterica increased their counts 1.05 and 1.11 log CFU cm2, respectively. Similar increases were obtained when incorporating bacteria to pre-adhered cells of Candida. Nevertheless C. krusei counts decreased in both experimental conditions, in a) 0.40 log CFU cm2 and 0.55 log CFU cm2 when exposed to E. coli O157:H7 and S. enterica and in b) 0.18 and 0.68 log CFU cm2, respectively. This suggests that C. krusei, E. coli O157:H7, and S. enterica have a complex relationship involving physical and chemical interactions on food contact surfaces. This study supports the possibility that pathogen interactions with members of spoilage microbiota, such as C. krusei, might play an important role for the survival and dissemination of E. coli O157:H7 and Salmonella enterica in food-processing environments. Based on the data obtained from the present study, much more attention should be given to prevent the contamination of these pathogens in acidic drinks.
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Adhesión Bacteriana , Candida/fisiología , Escherichia coli O157/fisiología , Jugos de Frutas y Vegetales/microbiología , Salmonella enterica/fisiología , Acero Inoxidable , Candida/aislamiento & purificación , Candida/ultraestructura , Medios de Cultivo/química , Escherichia coli O157/crecimiento & desarrollo , Escherichia coli O157/ultraestructura , Manipulación de Alimentos , Microbiología de Alimentos , Conservación de Alimentos , Malus/microbiología , Salmonella enterica/crecimiento & desarrollo , Salmonella enterica/ultraestructura , UltrafiltraciónRESUMEN
AIM: The occurrence of virulence markers, serotypes and invasive ability were investigated in Shiga toxin-producing Escherichia coli (STEC) isolated from faecal samples of healthy dairy cattle at Rio de Janeiro State, Brazil. METHODS AND RESULTS: From 1562 stx-positive faecal samples, 105 STEC strains were isolated by immuno-magnetic separation (IMS) or plating onto MacConkey agar (MC) followed by colony hybridisation. Fifty (47·6%) strains belonged to nine serotypes (O8:H19, O22:H8, O22:H16, O74:H42, O113:H21, O141:H21, O157:H7, O171:H2 and ONT:H21). The prevalent serotypes were O157:H7 (12·4%), O113:H21 (6·7%) and O8:H19 (5·7%). Virulence genes were identified by polymerase chain reaction (PCR). E-hlyA (77·1%) was the more prevalent virulence marker, followed by espP (64·8%), saa (39%), eae (24·8%) and astA (21·9%). All O157:H7 strains carried the γ (gamma) variant of the locus of enterocyte effacement (LEE) genes and the stx2c gene, while the stx1/stx2 genotype prevailed among the eae-negative strains. None of the eae-positive STEC produced the localized adherence (LA) phenotype in HEp-2 or Caco-2 cells. However, intimate attachment (judged by the fluorescent actin staining test) was detected in some eae-positive strains, both in HEp-2 (23·1%) and in Caco-2 cells (11·5%). Most strains (87·5%) showed 'peripheral association' (PA) adherence phenotype to undifferentiated Caco-2 cells. Twenty-five (92·6%) of 27 strains invaded Caco-2 cells. The highest average value of invasion (9·6%) was observed among the eae-negative bovine strains from serotypes described in human disease. CONCLUSION: Healthy dairy cattle is a reservoir of STEC carrying virulence genes and properties associated with human disease. SIGNIFICANCE AND IMPACT OF THE STUDY: Although reports of human disease associated with STEC are scarce in Brazil, the colonization of the animal reservoir by potentially pathogenic strains offers a significant risk to our population.
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Bovinos/microbiología , Reservorios de Enfermedades/microbiología , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Animales , Brasil , Células CACO-2 , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Heces/microbiología , Humanos , Reacción en Cadena de la Polimerasa , Serotipificación , Toxina Shiga/metabolismo , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/genética , Escherichia coli Shiga-Toxigénica/fisiología , Virulencia/genética , Factores de Virulencia/genética , Factores de Virulencia/metabolismoRESUMEN
The experiments were performed with Escherichia coli O157:H7 EDL 933 in freshwater microcosms at 12 °C. At 35, 45, and 70 days, samples were taken and filtered through 0.45 µm membrane filters. The following alternatives were tested to evaluate the recovery percentage of injured cells: (1) selective media CHROMagar(™)O157 and chromID(™)O157:H7 agar, at 37 °C for 24 h; (2) tryptic soy agar supplemented with yeast extract (TSAE), incubated at 25 °C for 2 or 4 h, then transferred to CHROMagar(™)O157 or chromID(™)O157:H7 agar at 37 °C (TSAE2h-CHROM, TSAE4h-CHROM and TSAE2h-ID, TSAE4h-ID); (3) thin agar layer (TAL) method, TSAE was overlaid on CHROMagar(™)O157 or chromID(™)O157:H7 agar (TALCHROM and TALID, respectively) and incubated at 37 °C for 24 h; and (4) TALCHROM at 25 °C for 4 h, then continued up to complete 24 h at 37 °C (TALCHROM4h). Furthermore, the recovery of E. coli O157:H7 cells adhering to glass coverslips were evaluated to mimic biofilm conditions. The recovery percentages obtained from each alternative were calculated relative to TSAE counts. After 70 days, TSAE4h-CHROM and TALCHROM4h showed the highest recovery percentage (>90 %) from water microcosms. Despite the improved recovery of cell adhering to glass surfaces, the percentages obtained with TSAE4h-CHROM were low. Further studies for the recovery of biofilm-forming E. coli O157:H7 are required. Pre-incubation on TSAE at 25 °C for 4 h, combined with CHROMagar(™)O157, or by thin agar layer method (TALCHROM) enhanced significantly the recovery of viable cells of E. coli O157:H7 after prolonged stay in water microcosms.
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Monitoreo del Ambiente/métodos , Escherichia coli O157/aislamiento & purificación , Agua Dulce/microbiología , Agua Subterránea/microbiología , Plancton/aislamiento & purificación , Agar , Biopelículas/crecimiento & desarrollo , Medios de Cultivo , Consorcios MicrobianosRESUMEN
Meat can be contaminated in different stages of the slaughtering process and the identification of these stages is the starting point to implement adequate control measures. The objectives of this study were to assess the presence of pathogenic microorganisms in cattle carcasses, to identify the most important contamination points of the slaughtering process, and to evaluate the possible risk factors related to them in a cattle slaughterhouse. To this aim, 108 cattle carcasses were sampled at three stages of the slaughtering process: Point 1 (hides after bleeding); Point 2 (carcasses after hide removal); and Point 3 (carcasses immediately after division). Escherichia coli O157:H7, Listeria monocytogenes and Salmonella Livingstone were isolated from the carcasses. Phenotypic and genotypic characterization indicated that there was cross-contamination among animals, since bacteria with identical genotypic and phenotypic profiles were isolated from different animals at the same sampling day. Furthermore, this is the first report about the isolation of E. coli O157:H7 in a bovine slaughterhouse from southern Brazil.
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Mataderos , Escherichia coli O157/aislamiento & purificación , Microbiología de Alimentos , Listeria monocytogenes/aislamiento & purificación , Carne/microbiología , Salmonella/aislamiento & purificación , Animales , Antibacterianos/farmacología , Brasil , Bovinos/microbiología , Farmacorresistencia Bacteriana , Contaminación de Equipos , Escherichia coli O157/efectos de los fármacos , Escherichia coli O157/genética , Genotipo , Listeria monocytogenes/efectos de los fármacos , Listeria monocytogenes/genética , Salmonella/efectos de los fármacos , Salmonella/genéticaRESUMEN
Pomegranate extract, vanillin and geraniol were studied as natural antimicrobials on strawberry juice. Strawberry juice was treated with each agent at two concentrations: pomegranate extract at 180 and 360 µg/mL; vanillin at 2.5 and 5 mg/mL; and geraniol at 0.6 and 1.2 µL/mL. After being treated, juices were stored at 5 °C and microbiological, physicochemical and sensory studies were carried out. Also, a second batch of juice was inoculated with Escherichia coli O157:H7 (10(5) CFU/mL) before being treated, to safety study. Geraniol and vanillin, at both concentrations tested, were highly effective in reducing the native microflora on strawberry juice (more than 3 log cycles), extending the microbiological shelf-life of the product. Moreover, both antimicrobials improved the product safety by reducing inoculated E. coli O157:H7. Furthermore, vanillin showed a significant increase in polyphenol content compared to untreated juice. On the other hand, pomegranate extract applied at the highest concentration showed important reductions on mesophilic and psychrophilic bacteria, but no effect on yeast and molds and inoculated E. coli. Even though vanillin and geraniol incorporation on strawberry juice had a negative effect on its sensory quality, pomegranate extract had no impact on the sensory attributes evaluated. Combinations of the biopreservatives could be studied in order to decrease the concentration of the antimicrobials, reducing the effects on strawberry juice sensory characteristics.
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BACKGROUND: Antimicrobial resistance in Shiga toxin-producing Escherichia coli (STEC) O157 and non-O157 is a matter of increasing concern. The aim of the present study was to investigate the antimicrobial resistance profiles of STEC O157 and non-O157 recovered from feces of domestic farm animals in the agricultural Culiacan Valley in Northwestern Mexico. FINDINGS: All of the examined STEC strains showed susceptibility to five antimicrobials, ceftazidime, ceftriaxone, ciprofloxacin, nalidixic acid, and trimethoprim-sulfamethoxazole. However, resistance to the four antimicrobials, ampicillin, cephalothin, chloramphenicol, and kanamycin was commonly observed. Interestingly, non-susceptibility to cephalothin was predominant among the examined STEC strains, corresponding to 85 % (22/26) of the O157:H7 from cattle, sheep and chicken and 73 % (24/33) of the non-O157 strains from cattle and sheep. Statistical analyses revealed that resistance to ampicillin was significantly correlated to 38 % (10/26) of STEC O157:H7 strains from multiple animal sources. Another significant correlation was found between serotype, source, and antimicrobial resistance; all of the O20:H4 strains, recovered from sheep, were highly resistant to tetracycline. Multidrug resistance profiles were identified in 42 % (22/53) of the non-susceptible STEC strains with clinically-relevant serotypes O8:H9, O75:H8, O146:H21, and O157:H7. CONCLUSIONS: STEC O157 and non-O157 strains, recovered from domestic farm animals in the Culiacan Valley, exhibited resistance to classes of antimicrobials commonly used in Mexico, such as aminoglycosides, tetracyclines, cephalosporins and penicillin but were susceptible to fluoroquinolones, quinolones, and sulfonamides. These findings provide fundamental information that would aid in the surveillance of antimicrobial resistance in an important agricultural region in Northwestern Mexico.
RESUMEN
Shiga toxin-producing Escherichia coli (STEC) are important food-borne pathogens, with the main virulence factor of this bacterium being its capacity to secrete Shiga toxins (Stxs). Therefore, the use of certain antibiotics for the treatment of this infection, which induces the liberation of Stxs, is controversial. Reactive oxygen and nitrogen species are also involved in the pathogenesis of different diseases. The purpose of this study was to analyze the effects of antibiotics on biofilms of STEC and the relationships between cellular stress and the release of Stx. To this end, biofilms of reference and clinical strains were treated with antibiotics (ciprofloxacin, fosfomycin and rifaximin) and the production of oxidants, the antioxidant defense system and toxin release were evaluated. Ciprofloxacin altered the prooxidant-antioxidant balance, with a decrease of oxidant metabolites and an increase of superoxide dismutase and catalase activity, being associated with high-levels of Stx production. Furthermore, inhibition of oxidative stress by exogenous antioxidants was correlated with a reduction in the liberation of Stx, indicating the participation of this phenomenon in the release of this toxin. In contrast, fosfomycin and rifaximin produced less alteration with a minimal production of Stx. Our data show that treatment of biofilm-STEC with these antibiotics induces oxidative stress-mediated release of Stx.
Asunto(s)
Antibacterianos/farmacología , Toxina Shiga I/metabolismo , Toxina Shiga II/metabolismo , Escherichia coli Shiga-Toxigénica/efectos de los fármacos , Animales , Antioxidantes/farmacología , Ácido Ascórbico/farmacología , Biopelículas , Catalasa/metabolismo , Supervivencia Celular/efectos de los fármacos , Chlorocebus aethiops , Ciprofloxacina/farmacología , Fosfomicina/farmacología , Glutatión/farmacología , Pruebas de Sensibilidad Microbiana , Óxido Nítrico/metabolismo , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Rifamicinas/farmacología , Rifaximina , Escherichia coli Shiga-Toxigénica/genética , Escherichia coli Shiga-Toxigénica/metabolismo , Escherichia coli Shiga-Toxigénica/fisiología , Superóxido Dismutasa/metabolismo , Células Vero , Factores de Virulencia/genéticaRESUMEN
Many attempts have been made to establish the control of foodborne pathogens through Lactobacillus isolates and their metabolism products with success being obtained in several situations. The aim of this study was to investigate the antagonistic effect of eight Lactobacillus isolates, including L. casei subsp. pseudoplantarum, L. plantarum, L. reuteri and L. delbrueckii subsp. delbrueckii, on the pathogenic Escherichia colistrain O157:H7. The inhibitory effect of pure cultures and two pooled cultures supernatants of Lactobacillus on the growth of pathogenic bacteria was evaluated by the spot agar method and by monitoring turbidity. Antimicrobial activity was confirmed for L. reuteri and L. delbrueckii subsp. delbrueckii and for a pool of lactic acid bacteria. The neutralized supernatant of the pool exerted a higher antimicrobial activity than that of the individual strains. Furthermore, D-lactic acid and acetic acid were produced during growth of the Lactobacillus isolates studied.(AU)
Muitas tentativas têm sido feitas para se estabelecer o controle de patógenos de origem alimentar através do uso de estirpes de Lactobacillus e dos seus produtos de metabolismo, com sucesso sendo sucedido em várias situações. O objetivo deste trabalho foi investigar o efeito antagônico do sobrenadante de culturas de oito isolados de Lactobacillus, incluindo L. casei subsp. pseudoplantarum, L. plantarum L. reuteri e L. delbrueckii subsp. delbrueckii, sobre Escherichia coli amostra O157:H7. Os efeitos inibidores de culturas puras e de dois "pools" de cultura de Lactobacillus sobre o crescimento da bactéria foram avaliados através do método de inibição em ágar e através do monitoramento da turbidez da cultura bacteriana. A atividade antimicrobiana foi confirmada para Lactobacillus reuteri e Lactobacillus delbrueckii subsp. delbrueckii e para o "pool" de bactérias acido-láctica. O sobrenadante neutralizado do "pool" de Lactobacillus exerceu uma atividade antimicrobiana mais elevada do que aquela das estirpes individuais. Além disso, ácido D-láctico e ácido acético foram produzidos durante o crescimento dos Lactobacillusestudados.(AU)
Asunto(s)
Animales , Lactobacillus/química , Lactobacillus/citología , Lactobacillus/inmunología , Escherichia coli O157/química , Escherichia coli O157/inmunologíaRESUMEN
Many attempts have been made to establish the control of foodborne pathogens through Lactobacillus isolates and their metabolism products with success being obtained in several situations. The aim of this study was to investigate the antagonistic effect of eight Lactobacillus isolates, including L. casei subsp. pseudoplantarum, L. plantarum, L. reuteri and L. delbrueckii subsp. delbrueckii, on the pathogenic Escherichia colistrain O157:H7. The inhibitory effect of pure cultures and two pooled cultures supernatants of Lactobacillus on the growth of pathogenic bacteria was evaluated by the spot agar method and by monitoring turbidity. Antimicrobial activity was confirmed for L. reuteri and L. delbrueckii subsp. delbrueckii and for a pool of lactic acid bacteria. The neutralized supernatant of the pool exerted a higher antimicrobial activity than that of the individual strains. Furthermore, D-lactic acid and acetic acid were produced during growth of the Lactobacillus isolates studied.(AU)
Muitas tentativas têm sido feitas para se estabelecer o controle de patógenos de origem alimentar através do uso de estirpes de Lactobacillus e dos seus produtos de metabolismo, com sucesso sendo sucedido em várias situações. O objetivo deste trabalho foi investigar o efeito antagônico do sobrenadante de culturas de oito isolados de Lactobacillus, incluindo L. casei subsp. pseudoplantarum, L. plantarum L. reuteri e L. delbrueckii subsp. delbrueckii, sobre Escherichia coli amostra O157:H7. Os efeitos inibidores de culturas puras e de dois "pools" de cultura de Lactobacillus sobre o crescimento da bactéria foram avaliados através do método de inibição em ágar e através do monitoramento da turbidez da cultura bacteriana. A atividade antimicrobiana foi confirmada para Lactobacillus reuteri e Lactobacillus delbrueckii subsp. delbrueckii e para o "pool" de bactérias acido-láctica. O sobrenadante neutralizado do "pool" de Lactobacillus exerceu uma atividade antimicrobiana mais elevada do que aquela das estirpes individuais. Além disso, ácido D-láctico e ácido acético foram produzidos durante o crescimento dos Lactobacillus estudados(AU)
Asunto(s)
Escherichia coli O157 , Ácido Acético/administración & dosificación , Ácido Láctico/administración & dosificación , Infecciones por Escherichia coli/prevención & control , LactobacillusRESUMEN
AIMS: The objective of this study was to determine the interactions between common spoilage yeast, Candida tropicalis, isolated from ultrafiltration membranes, and Escherichia coli O157:H7 and Salmonella sp. on stainless steel surfaces. METHODS AND RESULTS: Single and dual-species attachment assays were performed on stainless steel at 25°C using apple juice as culture medium. The growth of Salmonella sp. rose when it was co-cultivated with C. tropicalis in dual biofilms at 16 and 24 h; the same effect was observed for E. coli O157:H7 at 24 h. The colonization of C. tropicalis on stainless steel surfaces was reduced when it was co-cultivated with both pathogenic bacteria, reducing C. tropicalis population by at least 1.0 log unit. Visualization by SEM demonstrated that E. coli O157:H7 and Salmonella sp. adhere closely to hyphal elements using anchorage structures to attach to the surface and other cells. CONCLUSIONS: These results suggest a route for potential increased survival of pathogens in juice processing environments. These support the notion that the species involved interact in mixed yeast-bacteria communities favouring the development of bacteria over yeast. SIGNIFICANCE AND IMPACT OF THE STUDY: This study support the plausibility that pathogen interactions with strong biofilm forming members of spoilage microbiota, such as C. tropicalis, might play an important role for the survival and dissemination of E. coli O157:H7 and Salmonella sp. in food-processing environments.