Perfil microbilógico de diferentes tipos de saladas provenientes de cozinhas hospitalares

Pós-graduação em Medicina Veterinária - FMVZ

Qualidade microbiológica de alimentos servidos em restaurantes self-service

Collective food services have been increasing worldwide, and the self-service restaurant has been the current preference by consumers. Considering the importance of hygienic quality of food, the microbiological composition of ready-to-eat food was assessed. In the second semester of 2008, 20 samples of meals, mainly meat-based foods, were collected from different self-service restaurants in Araçatuba city, SP. Bacteriological analyses were performed following the conventional methodologies, and the results were compared with the standards established by the effective Brazilian legislation. Coliforms at 35ºC were detected in 90% of analyzed samples. Coliforms at 45°C were found in 55% of the samples and, among these, in 63.63%, the occurrence of Escherichia coli was confirmed. Coagulase-positive staphylococci were detected in 10% of samples and no sample showed Salmonella spp. or Bacillus cereus contamination. Sulfite reducing clostridia at 42o C were not investigated in this study. These findings indicate the need for a rigorous approach for improving the sanitary conditions during preparation and presentation of ready-to-eat food, as the consumption of contaminated products represents a potential risk to public health.

Evaluation of the use of probiotic acid lactic bacteria in the development of chicken hambúrguer

The objective of this study was to investigate the effect of fermentation with Lactobacillus acidophilus CRL 1014 on the physicochemical, microbiological and sensory characteristics of a hamburger product like processed with chicken meat and okara flour, with reduction of curing salts. A mixture of ingredients containing 90% chicken meat and 10% okara flour was subjected to the following treatments: F1: fermented with Lactobacillus acidophilus; F2:75 mg nitrite/kg and fermented with Lactobacillus acidophilus; F3: 150 mg nitrite/kg and unfermented. The quality of the “hamburgers” was assessed by physical and chemical analysis (pH, cooking yield and shrinkage), chemical composition, microbiological tests (Salmonella spp., count of sulphite-reducing clostridia, staphylococos coagulase-positive, total coliforms and Escherichia coli) and sensory analysis (sensory acceptance and purchase intent). During the first six days of fermentation, there was a decrease in pH from approximately 6.33 to 5.10. All the samples showed the same chemical composition (p < 0.05). The fermentation process was observed to inhibit the multiplication of microorganisms of several groups: coagulasepositive staphylococci, sulphite-reducing clostridia, Salmonella spp. and E. coli. The different “hamburgers” formulations showed high scores for all the sensory attributes evaluated, without differing from each other (p < 0.05). The results showed that the use of L. acidophilus CRL 1014 enabled the production of a safe product, with good physicochemical and sensory characteristics, in the absence of curing salts.

Assessment of the risk of raw milk consumption related to staphylococcal food poisoning

This study aimed to identify the risks of staphylococcal food poisoning due to the consumption of raw milk. Fifty-one farms in Londrina (PR) and 50 in Pelotas (RS) were analyzed, to determine the population of coagulase-positive staphylococci (UFC/ mL), as well as to verify the ability of producing Staphylococcal Enterotoxin A (SEA) by immunodifusion (OSP), the presence of the gene for the production of SEA (PCR) in the cultures, and the research of enterotoxin (SEA to SEE) in milk samples using ELISA commercial kit. Considering the 101 farms analyzed, 19 (18.8%) presented coagulase-positive staphylococci count above 105 UFC/mL. For the evaluation of the enterotoxigenic ability (SEA) by the OSP technique, six cultures coagulase-positive (5.5%) were positive to the test and identified as S. aureus. From the coagualse-negative sample, one (5.5%) was OSP positive. For the evaluation of the presence of the gene for EEA synthesis, 51 cultures of staphylococci were tested. From this total, 14 (27.45%) presented the gene, and from that, only 5 (9.81%) cultures were capable of expressing it in the technique of the OSP. The morphologic characteristic of the evaluated cultures that had enterotoxigenic capacity, from the 14 (33,3%) cultures that presented the gene for EEA production, 05 (11.9%) were characterized as typical cultures of S.aureus in Baird Parker agar. All the 12 milk samples studied for the presence of EEA to EEE in milk were negative. Thus, it can be concluded that there is extensive contamination of raw milk for staphylococci coagulase, however, most of the isolated strains were not enterotoxigenic or did not express such a characteristic. Only 9.81% of the tested colonies expressed the gene and effectively produced SEA. None of the samples had sufficient counts to produce detectable amounts of SEA. The milk samples did not present risk to cause staphylococcal food poisoning if consumed in natura until the collection moment.

Desenvolvimento de um embutido cárneo fermentado, com teores reduzidos de gordura e sais de cura, através da utilização de culturas probióticas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

EVALUATION OF MICROBIAL GROWTH ON SINGLE-USE VITRECTOMY PROBES REPROCESSED IN HEALTHCARE PRACTICE

The aim of this study was to evaluate the microbial growth on single-use vitrectomy probes reprocessed in healthcare practice. We investigated nine vitrectomy probes that had been reused and reprocessed using different methods. The samples were sectioned, individually, in portions of 3.5 cm, totaling 979 sampling units (extensions, connectors and vitrectomy cutters), which were inoculated in culture medium and incubated at 37 C for 14 days. The results showed microbial growth on 57 (5.8%) sample units, 25 of which had been sterilized using ethylene oxide, 16 by hydrogen peroxide plasma, and 16 by low-temperature steam and formaldehyde. Seventeen microbial species were identified. The most prevalent were: Micrococcus spp., coagulase-negative Staphylococcus, Pseudomonas spp., and Bacillus subtilis. The reuse of single-use vitrectomy probes was shown to be unsafe, therefore this practice is not recommended.

Comparison of Ozone and Chlorine in Low Concentrations as Sanitizing Agents of Chicken Carcasses in the Water Immersion Chiller

The aim of this study was to investigate the effects of the use of chlorine or ozone as sanitizing agents in the water of chicken immersion chilling, using the residual levels usually applied in Brazil (1.5 ppm), comparing the effects of these treatments on the microbiological, physicochemical, and sensory characteristics of carcasses. Chicken carcasses were chilled in water (4 degrees C) with similar residual levels of ozone and chlorine until reaching temperatures below 7 degrees C (around 45 min). The stability of carcasses was assessed during 15 days of storage at 2 +/- 1 degrees C. Microbiological, surface color (L*, a*, b* parameters), pH value, lipid oxidation (thiobarbituric acid reactive substances index), and sensory evaluation (on a 9-point hedonic scale for odor and appearance) analyses were carried out. The presence of Salmonella was not detected, coagulase-positive staphylococci counts were below 10(2) CFU/ml of rinse fluid, and Escherichia coil and total coliform counts were below 10(5) CFU/ml of rinse fluid until the end of the storage period for both treatments. Psychrotrophic microorganism counts did not differ (P > 0.05) between chlorine and ozone treatments, and both values were near 10(9) CFU/ml of rinse fluid after 15 days at 4 +/- 1 degrees C. pH values did not differ between treatments (P > 0.05) or during the storage period (P > 0.05). In addition, neither chlorine nor ozone treatment showed differences (P > 0.05) in the lipid oxidation of carcasses; however, the thiobarbituric acid reactive substances index of both treatments increased (P <= 0.05) during the storage period, reaching values of approximately 0.68 mg of malonaldehyde per kg. Samples from both treatments did not differ (P > 0.05) in their acceptance scores for odor and overall appearance, but in the evaluation of color, ozone showed an acceptance score significantly higher (P <= 0.05) than that for the chlorine treatment. In general, under the conditions tested, ozone showed results similar to the results for chlorine in the disinfection of chicken carcasses in the immersion chilling, which may indicate its use as a substitute for chlorine in poultry slaughterhouses.

Resistencia a antibióticos en bacterias aisladas de muestras de agua en Gran Canaria y su relación con la presencia de contaminantes emergentes

[ES]Hemos analizado la posible relación entre la presencia de contaminantes emergentes en agua regenerada para riego, agua del lixiviado del suelo y agua subterránea y la resistencia a antibióticos de las bacterias aisladas. Se tomaron muestras de agua. Cada muestra se dividió en dos partes. Una se centrifugó y se resembró en diferentes medios. Cincuenta mililitros de la segunda parte se añadieron a Caldo Brain Heart. En la siembra directa de agua de riego y solución del suelo se detectaron numerosas colonias. El crecimiento en la siembra directa de agua de galería fue escaso, y no hubo crecimiento cuando se sembró agua de pozo. Mediante enriquecimiento en Caldo BHI y resiembra posterior, se obtuvieron numerosas colonias

Impact of real time polymerase chain reaction technology on the therapeutic approach to treatment of staphylococcal infections

Can the early identification of the species of staphylococcus responsible for infection by the use of Real Time PCR technology influence the approach to the treatment of these infections? ^ This study was a retrospective cohort study in which two groups of patients were compared. The first group, ‘Physician Aware’ consisted of patients in whom physicians were informed of specific staphylococcal species and antibiotic sensitivity (using RT-PCR) at the time of notification of the gram stain. The second group, ‘Physician Unaware’ consisted of patients in whom treating physicians received the same information 24–72 hours later as a result of blood culture and antibiotic sensitivity determination. ^ The approach to treatment was compared between ‘Physician Aware’ and ‘Physician Unaware’ groups for three different microbiological diagnoses—namely MRSA, MSSA and no-SA (or coagulase negative Staphylococcus). ^ For a diagnosis of MRSA, the mean time interval to the initiation of Vancomycin therapy was 1.08 hours in the ‘Physician Aware’ group as compared to 5.84 hours in the ‘Physician Unaware’ group (p=0.34). ^ For a diagnosis of MSSA, the mean time interval to the initiation of specific anti-MSSA therapy with Nafcillin was 5.18 hours in the ‘Physician Aware’ group as compared to 49.8 hours in the ‘Physician Unaware’ group (p=0.007). Also, for the same diagnosis, the mean duration of empiric therapy in the ‘Physician Aware’ group was 19.68 hours as compared to 80.75 hours in the ‘Physician Unaware’ group (p=0.003) ^ For a diagnosis of no-SA or coagulase negative staphylococcus, the mean duration of empiric therapy was 35.65 hours in the ‘Physician Aware’ group as compared to 44.38 hours in the ‘Physician Unaware’ group (p=0.07). However, when treatment was considered a categorical variable and after exclusion of all cases where anti-MRS therapy was used for unrelated conditions, only 20 of 72 cases in the ‘Physician Aware’ group received treatment as compared to 48 of 106 cases in the ‘Physician Unaware’ group. ^ Conclusions. Earlier diagnosis of MRSA may not alter final treatment outcomes. However, earlier identification may lead to the earlier institution of measures to limit the spread of infection. The early diagnosis of MSSA infection, does lead to treatment with specific antibiotic therapy at an earlier stage of treatment. Also, the duration of empiric therapy is greatly reduced by early diagnosis. The early diagnosis of coagulase negative staphylococcal infection leads to a lower rate of unnecessary treatment for these infections as they are commonly considered contaminants. ^

Identification and bioactivity evaluation of two novel temporins from the skin secretion of the European edible frog, Pelophylax kl. Esculentus

The amphibian temporins, amongst the smallest antimicrobial peptides (AMPs), are α-helical, amphipathic, hydrophobic and cationic and are active mainly against Gram-positive bacteria but inactive or weakly active against Gram-negative bacteria. Here, we report two novel members of the temporin family, named temporin-1Ee (FLPVIAGVLSKLFamide) and temporin-1Re (FLPGLLAGLLamide), whose biosynthetic precursor structures were deduced from clones obtained from skin secretion-derived cDNA libraries of the European edible frog, Pelophylax kl. esculentus, by ‘shotgun’ cloning. Deduction of the molecular masses of each mature processed peptide from respective cloned cDNAs was used to locate respective molecules in reverse-phase HPLC fractions of secretion. Temporin-1Ee (MIC = 10 μM) and temporin-1Re (MIC = 60 μM) were both found to be active against Gram-positive Staphylococcus aureus, but retaining a weak haemolytic activity. To our knowledge, Single-site substitutions can dramatically change the spectrum of activity of a given temporin. Compared with temporine-1Ec, just one chemically-conservative substitution (Val8 instead of Leu8), temporin-1Ee bearing a net charge of +2 displays broad-spectrum activity with particularly high potency on the clinically relevant Gram-negative strains, Escherichia coli (MIC = 40 μM). These factors bode well for translating temporins to be potential drug candidates for the design of new and valuable anti-infective agents.

Melhoria da qualidade de jerked beef pela substituição de nitrito de sódio por extratos naturais de própolis e erva mate como agente antioxidante

Jerked beef, an industrial meat product obtained from beef with the addition of sodium chloride and curing salts and subjected to a maturing and drying process is a typical Brazilian product which has been gradually discovered by the consumer. The replacement of synthetic antioxidants by natural substances with antioxidant potential due to possible side effects discovered by lab tests, consumer health, is being implemented by the meat industry. This study aimed to evaluate the lipid oxidation of jerked beef throughout the storage period by replacing the sodium nitrite by natural extracts of propolis and Yerba Mate. For jerked beef processing brisket was used as raw material processed in 6 different formulations: formulation 1 (control - in nature), formulation 2 (sodium nitrite - NO), formulation 3 (Yerba Mate - EM), formulation 4 (propolis extract - PRO), formulation 5 (sodium nitrite + Yerba Mate - MS + NO), formulation 6 (propolis extract + sodium nitrite - PRO + NO). The raw material was subjected to wet salting, dry salting (tombos), drying at 25°C, packaging and storage in BOD 25°C. Samples of each formulation were taken every 7 days for analysis of lipid oxidation by the TBARS method. In all formulations, were carried out analysis of chemical composition at time zero and sixty days of storage. The water activity analysis and color (L *, a *, b *) was monitored at time zero, thirty and sixty days of storage. The Salmonella spp count, Coliform bacteria, Termotolerant coliforms and coagulase positive staphylococci were taken at time zero and sixty days. The activity of natural antioxidants evaluated shows the decline of lipid oxidation up to 2.5 times compared with the product in natura and presented values with no significant differences between treatments NO and EM, confirming the potential in minimize lipid oxidation of Jerked beef throughout the 60 days of storage. The results also showed that yerba mate has a higher antioxidant capacity compared to the propolis except the PRO + NO formulation. When associated with yerba mate with sodium nitrate, TBARS values become close to values obtained only for the control samples with the addition of sodium nitrite. The proximal composition of the formulations remained within the standards required in the IN nº22/2000 for jerked beef. Samples that differ significantly at 5% are directly related to the established type of formulation. The count of microorganisms was within the standards of the DRC nº12/2001 required for matured meat products. The intensity of the red (a*) decreased with storage time and increase the intensity of yellow (b*) indicates a darkening of the product despite L* also have been increased. These results suggest that yerba mate is a good alternative to meat industry in reducing healing addition salts when associated with another antioxidant.

Microbiological quality and safety of minimally processed fruits in the marketplace of southern Portugal

The availability of fresh-cut fruit (FCF) in the marketplace has been increasing in Portugal, although reports of its microbial quality are not known. Due to the growing concerns of these commodities over their microbial safety, the objectives of this work were to study the microbiological quality and prevalence of Salmonella and Listeria monocytogenes on fresh-cut fruits sold in southern Portugal. A study to examine the changes in pH and microbial counts, before and after the expiration dates, was also made. A total of 160 samples was purchased in the local grocery stores between September 2011 and August 2014, before their sell-by date. These samples were assayed for aerobic mesophilic (AM) and psychrotrophic (AP) microorganisms, yeasts and molds (YM), lactic-acid bacteria (LAB), coliforms (TC), Escherichia coli and coagulase positive staphylococci as well as L. monocytogenes and Salmonella. The microbiological counts ranged from 3.0-9.2 lg cfu/g (AM); 2.2–10.7 lg cfu/g (AP); 2.3–10.4 lg cfu/g (YM); 1.9–9.0 lg cfu/g (LAB) and less than 1–9.1 lg cfu/g (TC). The melons and watermelon presented the highest levels of the microbial quality parameters studied. However, no E. coli, staphylococci, Salmonella and L. monocytogenes were detected in any of the samples. After the sell-by date, an increase of the AM, AP, LAB and YM values was observed in all fruits. Conversely, the differences found in TC counts before and after the best-before date had no statistical significance. A decrease in pH was observed in all fruits except pineapple whose pH slightly increased after 14 days of storage. The results highlight the importance of preventing contamination and cross contamination, selecting adequate decontamination technologies and maintaining a strict temperature control during processing, distribution and selling of FCF.

Quality improvement of traditional dry fermented sausages based on innovative technological strategies

Dry fermented sausages are highly appreciated food specialties, mainly in Portugal and other southern European countries. Therefore, all research efforts aiming at improving the food quality and safety of traditional dry sausages are of interest, since they are likely to result in products with higher added value and quality standards most suited to the requirements and concerns of the modern consumers. Among those efforts, it may be highlighted the studies involving innovative processing parameters and technologies to overcome practical problems gathered in the meat industry, which are mostly associated with food quality and safety. Additionally, characterization of traditional dry sausages and rationalization of their processing are essential for further achievement of any official certification. Thus, this article attempts to point out some research lines of highest interest in meat science (and particularly to the broad variety of regional dry fermented sausages), towards to the valorisation of technological, nutritional and commercial features. In addition, it is here emphasized the importance for the continuous improvement of the quality and safety of meat products as a way to respond to the current concerns regarding its consumption and the general advices in reducing its daily intake.

Lack of horizontal gene transfer of methicillin-resitance genetic determinants from PBP2a-positive, coagulase-negative Staphylococci to methicillin-sensitive Staphylococcus aureus using transcutaneous electric nerve stimulation (TENS),

Previous research shows that approximately half of the coagulase-negative staphylococci (CNS) isolated from patients in the intensive care unit (ICU) at Belfast City Hospital were resistant to methicillin. The presence of this relatively high proportion of methicillin-resistance genetic material gives rise to speculation that these organisms may act as potential reservoirs of methicillinresistance genetic material to methicillin-sensitive Staphylococcus aureus (MSSA). Mechanisms of horizontal gene transfer from PBP2a-positive CNS to MSSA, potentially transforming MSSA to MRSA, aided by electroporation-type activities such as transcutaneous electrical nerve stimulation (TENS), should be considered. Methicillin-resistant CNS (MR-CNS) isolates are collected over a two-month period from a variety of clinical specimen types, particularly wound swabs. The species of all isolates are confirmed, as well as their resistance to oxacillin by standard disc diffusion assays. In addition, MSSA isolates are collected over the same period and confirmed as PBP2a-negative. Electroporation experiments are designed to mimic the time/voltage combinations used commonly in the clinical application of TENS. No transformed MRSA were isolated and all viable S. aureus cells remained susceptible to oxacillin and PBP2a-negative. Experiments using MSSA pre-exposed to sublethal concentrations of oxacillin (0.25 µg/mL) showed no evidence of methicillin gene transfer and the generation of an MRSA. The study showed no evidence of horizontal transfer of methicillin resistance genetic material from MR-CNS to MSSA. These data support the belief that TENS and the associated time/voltage combinations used do not increase conjugational transposons or facilitate horizontal gene transfer from MR-CNS to MSSA.

Determination of toxigenic capacity by reverse transcription polymerase chain reaction in coagulase-negative staphylococci and Staphylococcus aureus isolated from newborns in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)