Avaliação das boas práticas de fabricação e da contaminação de preparações cárneas de restaurantes institucionais e comerciais na cidade de Americana, SP / Evaluation of good manufacturing practices and contamination in meat preparations from institutional and commercial restaurants in the city of Americana/SP

Diante do ritmo acelerado da vida contemporânea, observa-se um aumento na tendência dos indivíduos em optar por realizar suas refeições fora de casa. A carne, reconhecida como um componente essencial na alimentação dos brasileiros, está suscetível à contaminação pois apresenta ambiente favorável à proliferação de microrganismos patogênicos. Fazendo-se necessária uma análise de contaminação pós-produção afim de evitar Doenças Transmitidas por Alimentos. No presente estudo objetivouse avaliar as boas práticas de fabricação e contaminação de preparações de carne bovina assada, de restaurantes particulares e institucionalizados no município de Americana-SP. Amostras de carne prontas para o consumo foram obtidas de seis estabelecimentos comerciais e seis institucionais. Durante a coleta, foram verificadas as temperaturas e realizadas análises de conformidades com a RDC n° 275, de 2002. As amostras foram examinadas para detectar a presença ou ausência de E. coli e coliformes termotolerantes a 45° C. Para a análise foi realizada a técnica de tubos múltiplos para quantificar a totalidade dos coliformes. Observou-se que, conforme estipulado pela Resolução n°43 de 2015, nenhuma das amostras oriundas de restaurantes comerciais, e a maioria das provenientes de restaurantes institucionais, atingiram as temperaturas requeridas. No que concerne à identificação de E. coli através de testes microbiológicos, foi constatado que seis amostras de restaurantes comerciais e quatro de restaurantes institucionais testaram positivo para a presença deste microrganismo. Conclui-se que as amostras de restaurantes comerciais apresentaram níveis de contaminação superiores em comparação com as amostras de restaurantes institucionais.

Given the fast-paced rhythm of contemporary life, there is an increase in individuals choosing to have their meals outside the home. Meat, recognized as an essential component in the Brazilian diet, is susceptible to contamination as it provides a favorable environment for the proliferation of pathogenic microorganisms. It is necessary to conduct post-production contamination analysis to prevent Foodborne Diseases. This study aimed to evaluate the good manufacturing practices and contamination of roasted beef preparations from private and institutional restaurants in the city of Americana-SP. Samples of ready-to-eat meat were obtained from six commercial establishments and six institutional ones. During collection, temperatures were checked, and conformity analyses were conducted according to RDC No. 275, 2002. The samples were examined for the presence or absence of E. coli and thermotolerant coliforms at 45°C using the multiple tube technique to quantify the total coliforms. It was observed that, as stipulated by Resolution No. 43, 2015, none of the samples from commercial restaurants and the majority from institutional restaurants reached the required temperatures. Regarding the identification of E. coli through microbiological tests, it was found that six samples from commercial restaurants and four from institutional ones tested positive for the presence of this microorganism. It is concluded that samples from commercial restaurants showed higher contamination levels compared to institutional restaurant samples.

Encapsulation of roast beef flavor by soy protein isolate/chitosan complex Pickering emulsions to improve its releasing properties during the processing of plant-based meat analogues.

During the production of plant-based meat analogues (PBMA), a significant loss of flavor characteristic compounds in meat-flavor essences could be observed. Pickering emulsion-based encapsulation is an effective method to improve their stability. Therefore, a soy protein isolate (SPI)/chitosan (CS) complex Pickering emulsion was fabricated to encapsulate roast beef flavor (RBF) and further applied in the processing of PBMA. Our results indicated that the network structure of emulsions was dominated by elasticity, while hydrogen and covalent bonding interactions played important roles in the encapsulation process. The release rate of flavor compounds gradually increased with the increase of pH value, glutamine transaminase, NaCl content, heating temperature or heating time, while encapsulation significantly reduced the loss of characteristic aroma compounds. In addition, the releasing characteristics of aroma compounds and textural properties of PBMA were greatly improved by treating with RBF-loaded emulsions. Consequently, the emulsions were promising to improve the flavor quality of PBMA.

Accumulation of Heterocyclic Amines and Advanced Glycation End Products in Various Processing Stages of Plant-Based Burgers by UHPLC-MS/MS.

The accumulation of heterocyclic amines (HAs) and advanced glycation end products (AGEs) during different processing stages was investigated in commercial raw materials to plant-based hamburger meats (PBHMs). Principal component analysis (PCA) was performed to explore the difference between the samples of each processing stage. The total free HA level accumulated from 4.74-6.63 ng/g in raw plant proteins to 5.81-20.23 ng/g in textured vegetable proteins after extrusion. The concentration of MeAαC increased from 29.23 ± 3.50 to 59.44 ± 0.26 ng/g, resulting in an accumulation of the total protein-bound HAs after cooking at 160 °C for 6 min, but the MeAαC content decreased to 42.26 ± 0.11 ng/g when the heating duration was prolonged to 12 min. An evident accumulation of AGEs was observed during the thermal home-processing of PBHM. The total levels for all HAs were 381.30 and 160.30 ng/g in roast beef patty (RBP) and PBHM, respectively, with RBP having a better amino acid composition pattern. These results may reveal the target processing stage, which should be paid attention to for the inhibition of Maillard reaction derivative harmful products (MRDHPs) in plant-based meat products.

Chitosan and flavonoid glycosides are promising combination partners for enhanced inhibition of heterocyclic amine formation in roast beef.

The effects of different kinds of chitosan, oligomer (ChiO) and monomer (Gluco), and the combinations of polymer (Chi) or ChiO with flavonoid aglycones and glycosides against the formation of major HAs were investigated to find out potential combination partners for enhanced suppression of HA formation. Results in roast beef patties showed ChiO and Gluco significantly inhibited PhIP and MeIQx formation by 43-80% and 31-57%, respectively. Of which, ChiO was the most effective. In combinations with flavonoid glycosides (phloridzin, rutin and hesperidzin, respectively), Chi, but not ChiO, generated enhanced inhibitory effects. Further analysis showed Chi and phloridzin combined at a ratio of 1:1 was the most promising, especially in inhibiting PhIP, and the mechanism behind involved: 1) water retention by Chi, and 2) reduction of phenylalanine availability by phloridzin. These findings suggest that appropriate combination of Chi and flavonoid glycosides contributes to significant improvement in the safety of meat products.

Toxicity Alleviation of Carbon Dots from Roast Beef after the Formation of Protein Coronas with Human Serum Albumin.

The unique properties of nanoparticles produced during food thermal processing have attracted considerable attention. In this study, the formation of protein coronas of fluorescent carbon dots (CDs) in roast beef with human serum albumin (HSA) and the corona effect on toxicity were reported. The CDs were roughly spherical with a size in the range of 1-5 nm, which were mainly composed of carbon (68.68%), nitrogen (10.6%), and oxygen (15.98%). The CDs could readily pass through the intestine wall due to their small size and good water solubility. There was an obvious interaction between HSA and CDs, suggesting that the CDs could form protein coronas. Thermodynamic analysis results of ΔH < 0 (-13.17 ± 3.74 kJ/mol) and ΔS > 0 ( 28.04 J/mol/K) indicated that the binding of HSA-CDs was due to electrostatic interactions or hydrophobic forces. The HSA-CD coronas were distributed in the lysosomes of the cells, alleviated swelling caused by the CDs, and inhibited the decrease of mitochondrial membrane potential caused by CDs. Furthermore, the protein coronas reduced cellular reactive oxygen species production and alleviated the consumption of glutathione by the CDs, thus protecting the cells from damage. This finding provided valuable information about protein coronas in ameliorating cytotoxicity.

Non-precursors amino acids can inhibit ß-carbolines through free radical scavenging pathways and competitive inhibition in roast beef patties and model food systems.

The aim of this study was to investigate the inhibitory effect of non-precursors amino acids (histidine, leucine, proline and methionine) which have advantages of safety, inexpensiveness and high standardization on the formation of ß-carbolines in roast beef patties and glucose/creatine/creatinine/tryptophan model system, and the possible pathway of inhibition by monitoring the scavenging of free radicals by electron paramagnetic resonance (EPR) spectroscopy and the consumption of tryptophan by HPLC in a glucose/tryptophan model system. Almost all amino acids can inhibit ß-carbolines in roast beef patties (up to 80.62%) and model system (up to 67.01%). Histidine showed an excellent alkyl radical scavenging ability (up to 82.59%) and a highly competitive inhibition ability (up to 65.60%) against ß-carbolines generation. The corresponding abilities of leucine and methionine were less remarkable. Proline could only suppress ß-carbolines through competitive inhibition. The results could shed light on the reduction of ß-carbolines during meat processing.

Assessment of Microbial Populations in the Manufacture of Vacuum-Packaged Ready-to-Eat Roast Beef and in a Related Production Plant.

Some microbiological criteria were monitored for 6 months in vacuum-packaged roast beef (15 production batches), raw beef (10 batches), and other meat products (12 batches) produced in an Italian small to medium-size enterprise. Fifty-five environmental swab samples also were analyzed. The main bacterial groups were identified by cultural methods according to International Organization for Standardization standards. Listeria monocytogenes was enumerated with the most-probable-number protocol, and species identification was confirmed with a specific PCR assay. Immediately after vacuum packaging, all ready-to-eat (RTE) products had low mean aerobic colony counts (<102 to 2.4 × 102 CFU g-1), anaerobic colony counts (1.6 to 6.5 × 101 CFU g-1), Enterobacteriaceae counts (1.1 to 1.4 × 101 CFU g-1), and Escherichia coli counts (generally below the detection limit). Nevertheless, the prevalence of L. monocytogenes in these samples was 3.7%. In roast beef samples, the aerobic and anaerobic colony counts reached unacceptable levels (>106 CFU g-1) after 14 days of refrigerated storage. Because the prevalence of L. monocytogenes increased to 13.3% during storage, a substantial reduction in the shelf life of these products is recommended. Surfaces without direct contact with food (floors and drains) had the highest mean counts for aerobic colonies (8.0 × 103 to 9.5 × 105 CFU/cm2), anaerobic colonies (2.9 × 103 to 3.2 × 104 CFU/cm2), Enterobacteriaceae (1.5 × 101 to 8.4 × 101 CFU/cm2), and E. coli (6.0 to 7.7 CFU/cm2). The levels of L. monocytogenes on direct food contact surfaces were below the detection limit, but more than 25% of floor samples were contaminated. These results reveal the persistence of L. monocytogenes in food processing environments, although at very low levels, posing a high risk of postcooking recontamination for RTE products. To improve hygienic conditions and reduce cross-contamination, an increase in operator awareness and a reassessment of surface sanitization protocols are needed.

Inhibition of Clostridium perfringens Growth during Extended Cooling of Cooked Uncured Roast Turkey and Roast Beef Using a Concentrated Buffered Vinegar Product and a Buffered Vinegar Product.

This research was conducted to evaluate the effectiveness of a concentrated buffered vinegar product (CBV) and a simple buffered vinegar product (BV) for controlling Clostridium perfringens outgrowth during extended cooling times of ready-to-eat roast turkey and roast beef. Whole turkey breasts and beef inside rounds were injected with a typical brine and then ground and mixed with CBV (0.0, 2.01, 2.70, and 3.30% [w/w]) or BV (0.0, 1.75, 2.25, and 3.75% [w/w]) and a three-strain C. perfringens spore cocktail to a detectable level of ca. 2 to 3 log CFU/g. The meat was divided into 10-g portions, vacuum packaged, and stored frozen until tested. The turkey and beef were cooked in a programmable water bath to 71.6°C (160.8°F) in 5 h and to 57.2°C (135°F) in 6 h, respectively. The cooked turkey and beef were then cooled exponentially from 48.9 to 12.8°C (120 and 55°F) in 6, 9, 12, 15, and 18 h for the five cooling treatments. The cooling continued until the temperature reached 4.4°C (40°F). C. perfringens counts were determined at 54.4°C (130°F) and 4.4°C. CBV at 2.01% effectively limited C. perfringens growth in turkey to ≤1 log CFU/g with up to a 9-h cooling treatment, and 2.70 and 3.30% solutions were effective with up to the 18-h cooling treatment. BV had an inhibitory effect on C. perfringens outgrowth in beef but did not limit growth to ≤1 log CFU/g at any concentration tested for any of the cooling treatments.

Formation of Free and Protein-Bound Heterocyclic Amines in Roast Beef Patties Assessed by UPLC-MS/MS.

The effect of different roasting temperatures on the amounts of 17 heterocyclic amines (HAs) from seven categories of both free and protein-bound states in roast beef patties was assessed using an ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. There were increased amounts and more types of HAs detected at higher roasting temperatures. Nine free HAs were detected at 250 °C, including PhIP (14.34 ± 0.36 ng/g), DMIP (1.02 ± 0.07 ng/g), 1,5,6-TMIP (1.70 ± 0.08 ng/g), MeIQ (0.36 ± 0.01 ng/g), IQx (0.37 ± 0.04 ng/g), MeIQx (9.94 ± 0.61 ng/g), 4,8-DiMeIQx (0.90 ± 0.05 ng/g), norharman (6.03 ± 0.30 ng/g), and harman (2.60 ± 0.09 ng/g). Also, 37.32 ng/g of total free HAs was generated. Twelve protein-bound HAs were detected in roast beef patties at 250 °C, including PhIP (1.70 ± 0.13 ng/g), DMIP (2.33 ± 0.25 ng/g), 1,5,6-TMIP (3.62 ± 0.49 ng/g), MeIQ (5.47 ± 0.18 ng/g), IQ[4,5-b] (0.70 ± 0.03 ng/g) MeIQx (4.03 ± 0.41 ng/g), 4,8-DiMeIQx (0.67 ± 0.09 ng/g), MeAαC (19.51 ± 1.12 ng/g), AαC (2.91 ± 0.45 ng/g), norharman (1304.96 ± 110.73 ng/g), harman (400.85 ± 25.29 ng/g), and Phe-P-1 (0.81 ± 0.06 ng/g). The highest amount of protein-bound HAs was 2913.31 ng/g at 175 °C. PhIP tended to exist in a free state, whereas MeIQ, harman, and norharman tended to exist in a protein-bound state. Furthermore, Phe-P-1, MeAαC, and AαC were detected only in a protein-bound state. These results could be useful for evaluating the exposure to HAs in a daily diet.

Survival and Metabolic Activity of Listeria monocytogenes on Ready-to-Eat Roast Beef Stored at 4 °C.

Three brands of commercial roast beef were purchased and artificially inoculated with a 5-strain Listeria monocytogenes cocktail at 2 inoculation levels (approximately 3 and 6 Log CFU/g). Although all 3 brands contained sodium diacetate and sodium lactate, inoculated Listeria cocktail survived for 16 d in all 3 brands; significant increases in L. monocytogenes numbers were seen on inoculated Brand B roast beef on days 12 and 16. Numbers of L. monocytogenes increased to 4.14 Log CFU/g for the 3 Log CFU/g inoculation level and increased to 7.99 Log CFU/g for the 6 Log CFU/g inoculation level by day 16, with the pH values being 5.4 and 5.8 respectively. To measure the cell viability in potential biofilms formed, an Alamar blue assay was conducted. Brand B meat homogenate had the highest metabolic activities (P < 0.05). By comparing its metabolic activities to Brands A and C and the inoculated autoclaved meat homogenates, results indicated that the microflora present in Brand B may be the reason for high metabolic activities. Based on the denaturing gradient gel electrophoresis and the Shannon-Wiener diversity index analysis, the "Brand" factor significantly impacted the diversity index (P = 0.012) and Brand B had the highest microflora diversity (Shannon index 1.636 ± 0.011). Based on this study, results showed that antimicrobials cannot completely inhibit the growth of L. monocytogenes in ready-to-eat roast beef. Native microflora (both diversity and abundance), together with product formula, pH, antimicrobial concentrations, and storage conditions may all impact the survival and growth of L. monocytogenes.

Application of a novel antimicrobial coating on roast beef for inactivation and inhibition of Listeria monocytogenes during storage.

The antilisterial efficacy of novel coating solutions made with organic acids, lauric arginate ester, and chitosan was evaluated in a three-stage study on inoculated roast beef for the first time. Ready-to-eat roast beef was specially ordered from the manufacturer. The meat surface was inoculated with five-strain Listeria monocytogenes cocktail inoculums at two different levels, ~3 and 6 Log CFU/cm(2) and treated with the stock solution (HAMS), the 1:5 diluted solution (MAMS), and the 1:10 diluted solution (LAMS) (stage 1). During the 20 min contact time, the antimicrobial coatings reduced the Listeria populations by approximately 0.9-0.3 Log CFU/cm(2). The higher the concentrations of the antimicrobial solution, the better the antilisterial effects were. The treated inoculated beef samples were then stored at 4 °C for 30 days. During storage, Listeria growth inhibition effects were seen. While no growth was seen from the HAMS-treated samples, a 1.6 Log CFU/cm(2) increase was seen for MAMS-treated samples, a 4.6 Log CFU/cm(2) increase was seen for LAMS-treated samples, and a 5.7 Log CFU/cm(2) increase was seen for NoAMS-treated samples on Day 30 (~3 Log CFU/cm(2) inoculation level). In the second stage, the impact of the roast beef storage time on solution's antilisterial effect was evaluated. Results showed that the effect of the antimicrobial solution was dependent on both the initial inoculation levels and storage times. In stage 3, the effect of the antimicrobial solution on roast beef quality was studied with both instrument measurement and sensory evaluation. Minor changes in color, pH, and water activity were found. However, only limited sensory differences were seen between the treated and untreated samples. When panels were able to accurately find color differences between samples, they preferred the treated samples. The findings of this research proved the antilisterial efficacy of the novel antimicrobial solution and showed its potential for being used as a roast beef cut surface coating to control Listeria contamination and for color protection.

Effects of Electrical Stimulation on Lipid Oxidation and Warmed-over Flavor of Precooked Roast Beef.

Many manufacturing processes damage the structure of meat products and this often contributes to lipid oxidation which could influence warmed-over flavor (WOF) in precooked beef that is reheated beef. Electrical stimulation causes contraction of muscles and improves tissue tenderization. The purpose of this study was to evaluate the rate of lipid oxidation or warmed-over flavor that could be affected by electrical stimulation of precooked roast beef after refrigerated storage and reheating. The results show that there was no significant difference between chemical compositions and cooking yields when comparing non-electrically stimulated and electrically stimulated roast beef. Moreover, electrical stimulation had no significant effect on oxidative stability and off-flavor problems of precooked roast beef as evaluated by thiobarbituric acid reactive substances (TBARS) and sensory test (warmed-over aroma and warmed-over flavor). However, there was an increased undesirable WOF and a decrease in tenderness for both ES and Non-ES treatments over refrigerated storage time. Electrical stimulation did cause reactions of amino acids or other compounds to decrease the desirable beef flavor in re-cooked meat.