Validation of bacterial growth inhibition models based on molecular properties of organic acids

Organic acids occur naturally in foods and have been used in many food products as preservatives because they inhibit the growth of most microorganisms. The acids commonly found in foods differ greatly in both their structure and inhibitory effects for different bacteria. A way to represent relationships between different acids was previously described in which principal components analysis (PCA) was applied to 11 physical and chemical properties of 17 organic acids, to arrive at principal properties. These were used for development of regression models that related the minimum inhibitory concentrations (MICs) of organic acids to their principal properties. Separate MIC models were constructed for six different bacteria. The objective of the present study was to test the predictive capabilities of the organism models using different organic acids from the ones used to construct the original models. MIC predictions were made for three acids for each of the six bacteria for which models were previously constructed. MIC determinations for these acids were then carried out and compared with the predictions; these were in good agreement, thus validating the models. The new data were combined with that obtained previously to produce similar, but slightly stronger models. These had R(2) values between 0.861 and 0.992.     

Behavior of Yersinia enterocolitica strains inoculated in model cheese treated with high hydrostatic pressure

The effects of high hydrostatic pressure treatment and the ability for survival, repair, and growth of three human pathogenic serotypes (O:1, O:3, O:8) of Yersinia enterocolitica were investigated in washed-curd model cheese made with pasteurized bovine milk. Samples were treated at 300, 400, and 500 MPa for 10 min at 20 degrees C and analyzed at 0, 1, 7, and 15 days to assess the viability of the Yersinia population. A long-term study (up to 60 days of ripening after high hydrostatic pressure treatment) was also undertaken. Treatments at 400 and 500 MPa caused maximum lethality, and only the treatment at 300 MPa showed significant differences (P < 0.05) between serotypes; the most baroresistant was O:3. Ability to repair and grow was not observed after 15 days of storage at 8 degrees C. Yersinia counts in untreated cheese samples also decreased below the detection limit at day 45 in the long-term study. These results suggest that the cheese environment did not allow recovery of injured cells or growth. A primary contributing factor to this effect seemed to be the low pH resulting from the production of lactic acid during cheese ripening.     

Determination of volatile compounds of precooked prawn (Penaeus vannamei) and cultured gilthead sea bream (Sparus aurata) stored in ice as possible spoilage markers using solid phase microextraction and gas chromatography/mass spectrometry

BACKGROUND: Solid phase microextraction (SPME) and gas chromatography/mass spectrometry (GC/MS) have been reported as useful techniques for analysing volatile compounds to monitor fish freshness. In this study, SPME/GC/MS was applied to cultured gilthead sea bream and precooked prawn stored in ice for 6 days in order to find possible markers of spoilage. Total volatile basic nitrogen (TVB‐N) was also determined as a common index of spoilage. RESULTS: The TVB‐N value at the end of the storage period for cultured gilthead sea bream (302.40 ± 8.50 mg kg^?1) was within the range of acceptability for edible fish (300–400 mg kg^?1) but could be considered at the beginning of spoilage. For precooked prawn the TVB‐N value at day 6 (863.04 ± 7.84 mg kg^?1) was not acceptable for human consumption. SPME/GC/MS identified 30 compounds in cultured gilthead sea bream and 49 compounds in precooked prawn. In particular, 3‐methyl‐1‐butanol, 2‐methylbutanal, 3‐methylbutanal and 3‐hydroxy‐2‐butanone increased during refrigerated storage both in the two species investigated here and in other species reported elsewhere and could be considered as markers of spoilage. CONCLUSION: This study confirmed that SPME/GC/MS can be considered an efficient method suitable for analysing the volatile compounds of both raw fish and fishery products in order to monitor loss of freshness. Copyright © 2008 Society of Chemical Industry     

Geographical Authentication of Tequila According to its Mineral Content by Means of Support Vector Machines

The elemental profile of tequila samples from the three main production areas of the State of Jalisco, namely Amatitlan, Guadalajara, and Tequila, was used to carry out their geographical characterization. With this aim, the concentration of Al, Ba, Ca, Cu, Fe, K, Mg, Mn, Na, S, Sr, and Zn was determined by inductively coupled plasma atomic emission spectroscopy. Principal component analysis was addressed to visualize data trends. The number of input variables was reduced by means of backward stepwise linear discriminant analysis and support vector machines were used to construct an adequate classification model. The best classification performance was obtained by a linear support vector machine model with 100% of prediction ability.     

Characterization of Edible Active Coating Based on Alginate–Thyme Oil–Propionic Acid for the Preservation of Fresh Chicken Breast Fillets

Chicken meat presents serious problems of processing and storage. Edible coatings are used commercially to improve the shelf life of fresh foods. The aim of this study is to develop an effective antimicrobial edible coating to improve the shelf life and safety of fresh chicken meat. The effect of propionic acid and thyme essential oils as antimicrobial and antioxidant compounds incorporated into alginate‐based edible coating was evaluated. Physical, antimicrobial and sensorial analyses were performed in coated and uncoated samples. Sensorial analysis showed no significant differences between coated and uncoated samples, which do not influence the buying decisions of consumers. The pH of the meat surface, color and sensorial analysis changed during storage time, showing similar behavior between coatings, the principal parameters to determine the shelf life of this product were weight loss and microbiological deterioration, where coatings had different behaviors. The selected coating increased the shelf life by about 33% with the lowest dehydration.     

Mineral profile of “fino” wines using inductively coupled plasma optical emission spectrometry methods

Fifty wines from the Denomination of origin (DO) of Condado de Huelva were analysed for mineral content by measuring 12 elements (Al, Ba, Ca, Cu, Fe, K, Mg, Mn, Na, P, Sr and Zn) using Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES). Samples were previously digested by heating with H₂O₂/HNO₃ mixture. The results obtained showed that metal data set were non-normally distributed and accordingly, non-parametric statistics were applied. The average levels (medians) of these elements found in the samples are as follows, in mg/L: 2.54 (Al); 0.06 (Ba); 82.58 (Ca); 0.21 (Cu); 3.53 (Fe); 865.34 (K); 68.87 (Mg); 0.71 (Mn); 32.77 (Na); 71.61 (P); 0.48 (Sr); 0.56 (Zn). The interrelation of metal couples was studied through the Spearman non-parametric sample correlation, being Fe/Al, P/Mg, and Zn/Ba the most important correlations established. As a result of this study, we can suggest that the contribution to the safety intake limits (per week) of the studied elements through the wine consumption is not significant. Actually, they range between 0.1% in Fe and 11.9% in Mg, for normal drinkers.     

Composition of meat and offal from weaned and fattened rabbits and results of stereospecific analysis of triacylglycerols and phosphatidylcholines

BACKGROUND: Rabbit meat has excellent nutritive properties. The purpose of this study was to characterize rabbit meat and offal; in particular, the lipid fraction was studied in order to evaluate total and positional fatty acid (FA) compositions of triacylglycerol (TAG) and phosphatidylcholine (PC) fractions. Eight samples of weaned and eight of fattened rabbits were considered. RESULTS: Fattened rabbit meat contained slightly higher protein percentage content (P < 0.05) in comparison to weaned (20.1% versus 18.0%). Calcium content was higher in meat than in offal, unlike sodium, iron, zinc, manganese and copper. The cholesterol content in offal was much higher than in meat. FA profiles of total lipid showed a high percentage of unsaturated fatty acids and an n‐6/n‐3 ratio of 10.3 for fattened rabbit meat. Stereospecific analysis of TAG and PC was carried out on an eight‐sample pool of each meat and offal from weaned and fattened rabbits. In all samples the sn‐2‐position was prevalently esterified with oleic and linoleic acids in TAG, with polyunsaturated fatty acids in PC. CONCLUSION: Lipids from rabbit meat presented higher content of monounsaturated FA and lower n‐6/n‐3 ratio in comparison to offal, which was characterized by higher cholesterol and mineral levels. Copyright © 2011 Society of Chemical Industry     

Cloning, characterization, expression and antifungal activity of an alkaline serine protease of Aureobasidium pullulans PL5 involved in the biological control of postharvest pathogens

An alkaline protease gene was amplified from genomic DNA and cDNA of the antagonistic yeast-like fungus Aureobasidium pullulans PL5, a biocontrol agent effective against Monilinia laxa on stone fruit and Botrytis cinerea and Penicillium expansum on pome fruits. An open reading frame of 1248 bp encoding a 415-amino acid (aa) protein with a calculated molecular weight (M_r) of 42.9 kDa and an isoelectric point (pI) of 4.5 was characterized. The cDNAALP5 gene had an 18-amino acid signal peptide, one N-gylcosylation, one histidine active site, and one serine active site. The ALP5 gene with a M_r of 1351 bp contained two introns. One intron was of 54 bp, while the other was of 50 bp. Protein BLAST and phylogenetic tree analysis of the deduced amino sequences from the cDNAALP5 gene showed that the encoded protein had 100% homology to a protease enzyme (ALP2) of a sea strain of A. pullulans, suggesting that the protein ALP5 was an alkaline serine protease. Expression of ALP5 in Escherichia coli BL21 (DE3), followed by identification with Western-blotting, purification with Ni-NTA and analysis of enzymatic activity, yielded an homogeneous recombinant ALP5 which hydrolysed the substrate casein and inhibited the mycelial growth of the pathogens. At its optimal pH of 10.0 and reaction temperature of 50 °C, the recombinant protease exhibited the highest activity towards the substrate casein, though the highest stability was at lower temperatures and pH between 7.0 and 9.0. This study provided the direct evidence that extracellular proteases secreted by the antagonist A. pullulans PL5 played a role in the biocontrol activities against some postharvest pathogens of apple and peach.     

Listeria monocytogenes in Gorgonzola: subtypes, diversity and persistence over time

L. monocytogenes represents a primary concern in the production of Gorgonzola, a Protected Designation of Origin (PDO) Italian blue-veined cheese produced only in the Piedmont and Lombardy regions. L. monocytogenes isolates (N=95) obtained from Gorgonzola rinds, paste, and production/ripening environments were serotyped and then genotyped using Pulsed Field Gel Electrophoresis (PFGE). The goal of this study was to investigate the variability of L. monocytogenes PFGE-types across different PDO Gorgonzola manufacturers (N=22). The majority of the strains (88%) were serotyped as 1/2a. PFGE identified 2 major pulse-types grouping 62 strains, detected from different plants and years, suggesting the presence of persistent and niche-adapted L. monocytogenes. In 9 plants, environmental strains shared the same pulse-types with strains from rinds or paste, suggesting a possible transmission pathway. Encouragingly, L. monocytogenes was retrieved from only 1 paste, indicating that production processes were under control in 21 plants. In the remaining plant, un-effective pasteurization or cross-contamination during production processes could be the cause of the contamination. Consequently, it is imperative that producers operate under the total respect of the Good Manufacturing Practices and following the principles of the Hazard Analysis Critical Control Point plans, in order to contain contamination throughout the whole processing.