RESUMEN
The impact of primary cooling on beef microbiota was investigated on six beef carcasses consecutively processed with the parallel use of metataxonomic and culture-dependent analysis. Samples were collected immediately after slaughtering (AS) and after the 24th-hour post-cooling (PC) from three different surfaces, namely neck, flank and thigh. The main objective was to examine whether the microbiota composition of beef carcasses changes as function of the surface sampled, primary cooling (from AS to PC) and animal's origin (breeder). The outcomes underline that primary cooling did not affect qualitatively the composition of the potentially active microbiota or the carcass superficial counts. Although slight changes in chemical-physical parameters like volatile organic compounds (VOCs) were observed after cooling, the carcasses microbiota and its inferred metabolic pathways varied among animals as a function of their origin. Co-occurrence and co-exclusion analyses underlined competition for the colonisation of the carcass surface between Brochothrix-Psychrobacter and Carnobacterium-Serratia-Pseudomonas. Once integrated in a comprehensive monitoring of the supply chain, the metataxonomic characterisation of the beef carcasses microbiota might represent a valid integrative approach to define the cuts' perishability and their appropriateness to specific packaging and storage methods. These new bits of knowledge could be the base to define good strategies for the prevention of meat spoilage.
Asunto(s)
Microbiología de Alimentos , Microbiota , Animales , Bovinos , Carne , FríoRESUMEN
A low initial contamination level of the meat surface is the sine qua non to extend the subsequent shelf life of ground beef for as long as possible. Therefore, the short- and long-term effects of a pregrinding treatment with electrolyzed water (EW) on the microbiological and physicochemical features of Piedmontese steak tartare were here assessed on site, by following two production runs through storage under vacuum packaging conditions at 4°C. The immersion of muscle meat in EW solution at 100 ppm of free active chlorine for 90 s produced an initial surface decontamination with no side effects or compositional modifications, except for an external color change that was subsequently masked by the grinding step. However, the initially measured decontamination was no longer detectable in ground beef, perhaps due to a quick recovery by bacteria during the grinding step from the transient oxidative stress induced by the EW. We observed different RNA-based metataxonomic profiles and metabolomic biomarkers (volatile organic compounds [VOCs], free amino acids [FAA], and biogenic amines [BA]) between production runs. Interestingly, the potentially active microbiota of the meat from each production run, investigated through operational taxonomic unit (OTU)-, oligotyping-, and amplicon sequence variant (ASV)-based bioinformatic pipelines, differed as soon as the early stages of storage, whereas microbial counts and biomarker dynamics were significantly distinguishable only after the expiration date. Higher diversity, richness, and abundance of Streptococcus organisms were identified as the main indicators of the faster spoilage observed in one of the two production runs, while Lactococcus piscium development was the main marker of shelf life end in both production runs. IMPORTANCE Treatment with EW prior to grinding did not result in an effective intervention to prolong the shelf life of Piedmontese steak tartare. Our RNA-based approach clearly highlighted a microbiota that changed markedly between production runs but little during the first shelf life stages. Under these conditions, an early metataxonomic profiling might provide the best prediction of the microbiological fate of each batch of the product.
Asunto(s)
Contaminación de Alimentos/análisis , Lactococcus/crecimiento & desarrollo , Microbiota/efectos de los fármacos , Carne Roja/microbiología , Streptococcus/crecimiento & desarrollo , Agua/farmacología , Animales , Bovinos , Manipulación de Alimentos/métodos , Microbiología de Alimentos , Almacenamiento de Alimentos/métodos , Lactococcus/efectos de los fármacos , Lactococcus/aislamiento & purificación , Streptococcus/efectos de los fármacos , Streptococcus/aislamiento & purificación , Agua/química , Microbiología del AguaRESUMEN
Fungi have a crucial role in the correct maturation of salami, but special attention should be addressed to the production of the nephrotoxic, immunotoxic, and carcinogenic mycotoxin ochratoxin A (OTA). In a monitoring study conducted in Northern Italy, OTA was detected by liquid chromatography coupled with mass spectrometry in 13 out 133 samples of traditional salami (9.8% of the total count). Mycological analysis of these samples yielded 247 fungal isolates which were identified to species level. The most frequent species were Penicillium nalgiovense, P. solitum, and P. chrysogenum. P. nordicum, an OTA-producing species commonly found in proteinaceous food, was not found in these samples. Three isolates were found to be Aspergillus westerdijkiae, an OTA-producing species. In order to check the results of the microbiological identification, 19 different strains of Aspergillus and 94 of Penicillium were tested for the presence of a sequence common to OTA-producing fungi by real-time PCR. None of the studied isolates, including the three A. westerdijkiae, possessed the otanpsPN target which is common to OTA-producing strains. Two out of three isolates of the A. westerdijkiae were also PCR-negative for the otanpsPN gene and did not produce OTA in culture. Conversely, this target sequence was amplified from the DNA purified from 14 salami casings including three casings harboring A. westerdijkiae. The amplification of sequences specific for OTA-producing strains performed on total genomic DNA extracted directly from salami casings provided a more suitable approach than PCR analysis of isolates from salami for the OTA-related otanpsPN gene to evaluate the risk of OTA contamination.
Asunto(s)
Análisis de los Alimentos , Contaminación de Alimentos , Microbiología de Alimentos , Hongos/metabolismo , Ocratoxinas/análisis , Cromatografía Liquida , ADN Espaciador Ribosómico , Análisis de los Alimentos/métodos , Hongos/clasificación , Hongos/genética , Hongos/aislamiento & purificación , Italia , Microbiota , Micotoxinas/análisis , Micotoxinas/biosíntesis , Micotoxinas/genética , Ocratoxinas/biosíntesis , Penicillium/clasificación , Penicillium/genética , Penicillium/aislamiento & purificación , Penicillium/metabolismo , Reacción en Cadena de la Polimerasa , Espectrometría de Masas en TándemRESUMEN
The aim was to study how factors such as temperature, alcoholic degree, and amino acids supplementation are able to influence the content of tyramine, histamine, 2-phenylethylamine, tryptamine and their precursor amino acids in winemaking process. Biogenic amines and amino acids were quantified at the beginning, middle and end of alcoholic fermentation, and at the end of malolactic fermentation. In general, samples produced with amino acid supplementation did not show the highest concentrations of biogenic amines, except for histamine, which content increased with the addition of the four amino acids. The synthesis of tyramine was mainly affected by the temperature and alcoholic degree, the formation of phenylethylamine was largely influenced by alcoholic degree, and tryptamine synthesis principally depended on temperature. Interestingly, there was interaction between these three factors for the biogenic amines studied. In conclusion, winemaking conditions should be established depending on the biogenic amine which synthesis is required to be controlled.
Asunto(s)
Aminoácidos/análisis , Aminas Biogénicas/análisis , Etanol/análisis , Temperatura , Vino/análisis , Fermentación , Histamina/análisis , Fenetilaminas/análisis , Triptaminas/análisis , Tiramina/análisisRESUMEN
The validation of a HPLC-PDA-MS/MS chromatographic method for the quali/quantitative characterization of histaminol, hydroxytyrosol, tyrosol, and tryptophol in wine has been described and discussed. Four standards showed a good linearity with high correlation coefficient values (over 0.9989) and LOD and LOQ were 0.001-0.015 mg/L and 0.004-0.045 mg/L, respectively. Furthermore, this study reported how factors such as temperature, alcoholic degree, and amino acids concentration are able to influence the formation of these four alcohols in Monastrell wines. The quantification values of these alcohols has been detected both at the half and end of alcoholic fermentation, and at the end of malolactic fermentation. In relation to interactions between factors, several significant variations emerged (p ⩽ 0.001). The impact of amino acids supplementation in Monastrell must it has been demonstrated, mainly in regards to histaminol and tryptophol.
Asunto(s)
Fermentación , Histamina/análogos & derivados , Indoles/análisis , Alcohol Feniletílico/análogos & derivados , Vino/análisis , Aminoácidos/análisis , Cromatografía Líquida de Alta Presión , Histamina/análisis , Histamina/biosíntesis , Alcohol Feniletílico/análisis , Espectrometría de Masas en Tándem , TemperaturaRESUMEN
According to European Union Regulation EC 1531/2001, olive oil labelled as "extra-virgin" should be cold-pressed and contain no refined oil or oil from other oleaginous seeds or nuts. Adulteration of extra virgin olive oil (EVOO) with hazelnut oil (HAO) is a serious concern both for oil suppliers and consumers. The high degree of similarity between the two fats complicates the detection of low percentages of HAO in EVOO. Many analytical approaches have been developed in recent years to trace HAO in EVOO, principally based on chromatographic analyses, differential scanning calorimetry or nuclear magnetic resonance. In addition adulteration of EVOO with HAO may introduce hazelnut-derived allergens. The aim of this work was to analyse the protein and allergen content of EVOO intentionally spiked with raw cold-pressed HAO or solvent-extracted HAO. SDS-PAGE analysis confirmed the presence of hazelnut proteins in solvent-extracted HAO with molecular masses ranging 10-60 kDa. In contrast, cold-pressed HAO showed no traces of protein. In spiked EVOO, solvent-extracted HAO was still detectable at a 1% contamination level. Several bands on SDS-PAGE migrated at apparent molecular masses coinciding with known allergens, such as Cor a 1 (approximately 17 kDa), Cor a 2 (approximately 14 kDa), Cor a 8 (approximately 12 kDa), oleosin (approximately 17 kDa) and Cor a 9 (approximately 60 kDa). MALDI-TOF MS analysis confirmed the presence of two oleosin isoforms and of Cor a 9. Immunoblotting demonstrated that an allergic patient with known reactivity to Cor a 1 and Cor a 2 recognized a 17-kDa band in solvent-extracted HAO. In conclusion, we have shown that adulteration of extra virgin olive oil with solvent-extracted hazelnut oil can be traced by simple SDS-PAGE analysis, and that adulteration introduces a potential risk for hazelnut allergic patients.
Asunto(s)
Corylus/efectos adversos , Corylus/inmunología , Grasas Insaturadas en la Dieta/efectos adversos , Contaminación de Alimentos/análisis , Hipersensibilidad a los Alimentos/etiología , Aceites de Plantas/efectos adversos , Alérgenos/análisis , Alérgenos/genética , Secuencia de Aminoácidos , Corylus/química , Corylus/genética , Grasas Insaturadas en la Dieta/análisis , Electroforesis en Gel de Poliacrilamida , Hipersensibilidad a los Alimentos/inmunología , Humanos , Aceite de Oliva , Aceites de Plantas/análisis , Proteínas de Plantas/efectos adversos , Proteínas de Plantas/análisis , Proteínas de Plantas/genética , Proteínas de Plantas/inmunología , Factores de Riesgo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Wine, like other fermented foods, may contain biogenic amines produced by lactic acid bacteria via amino acids decarboxylation. The most relevant amines from the toxicological standpoint are histamine and tyramine. The complexity of fermented substrates makes it difficult to suggest a priori how variables can modulate amine production. Lactobacillus hilgardii ISE 5211 was isolated from an Italian red wine. Besides producing lactate from malate, this strain is also able to convert arginine to ornithine and histidine to histamine. In the present investigation we studied the influence of malate, arginine and ethanol on histamine accumulation by L. hilgardii ISE 5211. Ethanol concentrations above 13% inhibit both histamine accumulation and bacterial growth; concentrations below 9% affect neither growth nor histamine production. However, an ethanol concentration of 11% allows a low but continuous accumulation of histamine to occur. Arginine also delays histamine accumulation, while malate appears to have no effect on histidine-histamine conversion.