Improvement of shelf-life of cherry (Prunus avium L.) by combined application of modified-atmosphere packaging and antagonistic yeast for long-distance export.

BACKGROUND: The last decade has seen a growing interest in reducing the use of chemical fungicides for postharvest decay control. In the research for new, safe alternatives, the combined application of biocontrol agents and passive modified-atmosphere packaging (MAP) has been shown to be a promising strategy to extend fruit quality. Therefore, the aim of this work was to evaluate the effect of the combined application of MAP and two antagonistic yeasts, Metschnikowia pulcherrima L672 and Pichia kudriavzevii PK18, on sweet cherry shelf life. RESULTS: Microbiological, physico-chemical, and quality fruit analysis from batches treated with antagonistic yeast were compared with a control batch without yeast application and a batch to which fludioxonil (Scholar®) was applied. The composition of the atmosphere and physico-chemical traits showed similar values among the different batches during cold storage. However, interestingly, the combination of MAP with the antagonistic yeasts M. pulcherrima L672 and P. kudriavzevii PK18 increases the control of microbiological spoilage with results comparable to the application of fludioxonil. In addition, these batches experienced a slight decrease in volatile compounds associated with fresh fruit aroma, whereas in the control batch an increase of altered fruit aromas was observed. The same effect of control of spoilage was observed during the shelf life period. CONCLUSION: These results showed the positive effect of the combination of antagonistic yeasts and MAP, obtaining similar results in terms of control of microbiological spoilage and physico-chemical quality compared with the application of fludioxonil. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Antimicrobial Properties of Essential Oils Obtained from Autochthonous Aromatic Plants.

The aim of this work was to determine the antimicrobial activity of the essential oils of six plants widely distributed in the Dehesa of Extremadura, such as Calendula officinalis, Cistus ladanifer, Cistus salviifolius, Cistus multiflorus, Lavandula stoechas, and Rosmarinus officinalis. The content of total phenolic compounds (TPC) and the antimicrobial activity of the essential oils against pathogenic and spoilage bacteria and yeasts as well as aflatoxin-producing molds were determined. A great variability was observed in the composition of the essential oils obtained from the six aromatic plants. The Cistus ladanifer essential oil had the highest content of total phenols (287.32 ppm), followed by the Cistus salviifolius essential oil; and the Rosmarinus officinalis essential oil showed the lowest amount of these compounds. The essential oils showed inhibitory effects on the tested bacteria and also yeasts, showing a maximum inhibition diameter of 11.50 mm for Salmonella choleraesuis and Kregervanrija fluxuum in the case of Cistus ladanifer and a maximum diameter of 9 mm for Bacillus cereus and 9.50 mm for Priceomyces carsonii in the case of Cistus salviifolius. The results stated that antibacterial and antiyeast activity is influenced by the concentration and the plant material used for essential oil preparation. In molds, aflatoxin production was inhibited by all the essential oils, especially the essential oils of Cistus ladanifer and Cistus salviifolius. Therefore, it can be concluded that the essential oils of native plants have significant antimicrobial properties against pathogenic and spoilage microorganisms, so they could be studied for their use in the industry as they are cheap, available, and non-toxic plants that favor the sustainability of the environment of the Dehesa of Extremeña.

Control of toxigenic Aspergillus spp. in dried figs by volatile organic compounds (VOCs) from antagonistic yeasts.

Aspergillus flavus and Aspergillus niger are fungi which can contaminate dried figs before and after harvest and consequently produce aflatoxins (AFs) and ochratoxin A (OTA). Many approaches have been applied to minimise the growth of these filamentous fungi, mainly involving the use of synthetic fungicides which are limited due to their negative impact on human health and the environment. In this context, biocontrol is a recent approach that needs to be explored. This study evaluated the potential of three volatile organic compounds (VOCs), octanoic acid (OA), 2-phenylethyl acetate (2PEA) and furfuryl acetate (FA), produced by Hanseniaspora uvarum and Hanseniaspora opuntiae yeasts on the growth, germination, gene expression and production of AFs and OTA by A. flavus M144 and A. niger M185 on dried fig-based agar and the incidence rates in dried figs. Two of the three VOCs evaluated (2PEA and FA) effectively controlled A. flavus M144 and A. niger M185 by using at least amounts of 50 µL (715 µL/L in the headspace) for FA and 100 µL (1430 µL/L in the headspace) for 2PEA in dried figs. One of the mode of actions of both compounds consists in early repressing the expression of genes involved in the biosynthesis of AFs (aflR) and OTA (pks) of A. flavus and A. niger, respectively. The results of this study support the application of 2PEA and FA at the early post-harvest stages of dried figs to control mycotoxin accumulation.

Retinol and α-Tocopherol Contents, Fat Color, and Lipid Oxidation as Traceability Tools of the Feeding System in Suckling Payoya Kids.

The effects of Payoya kid feeding systems on the fat-soluble vitamin (retinol/α-tocopherol) contents, fat content, fat color, and the oxidation index were evaluated to determine their potential for use as feeding system traceability tools. Four groups of Payoya kids (55 animals in total) fed milk exclusively were studied: a group fed a milk replacer (MR) and three groups fed natural milk from dams reared with different management systems (mountain grazing (MG), cultivated meadow (CM) and total mixed ration (TMR)). Kids were slaughtered around one month of age and 8 kg of live weight. Kids from the MG and CM groups presented lower retinol (5.56 and 3.72 µg/mL) and higher α-tocopherol plasma (11.43 and 8.85 µg/mL) concentrations than those from the TMR and MR groups (14.98 and 22.47 µg/mL of retinol; 2.49 and 0.52 µg/mL of α-tocopherol, respectively) (p < 0.001). With respect to fat, kids with a higher intramuscular fat percentage (CM and TMR groups) had lower retinol contents (16.52 and 15.99 µg/mL, respectively) than kids from the MG and MR groups (26.81 and 22.63 µg/mL, respectively) (p < 0.001). A dilution effect of vitamins on fat was shown: the higher the amount of fat, the lower the vitamin concentrations, the higher the lipid oxidation index (MDA), and the lower the SUM (absolute value of the integral of the translated spectra between 450 and 510 nm). A discriminant analysis that included all studied variables showed that 94.4% of the kids were classified correctly according to their feeding system and could allow traceability to the consumer.

Identification of the Causal Agent of Aqueous Spot Disease of Sweet Cherries (Prunus avium L.) from the Jerte Valley (Cáceres, Spain).

The pre and postharvest disease named 'aqueous spot' is an emerging risk for sweet cherries growing in Jerte Valley (Cáceres, Spain). Early stages of the disease appear in the tree, but it is usually detected after harvesting, during the postharvest period. Symptoms include the appearance of skin discolouration and translucency in the shoulder areas. At the most advanced stages, a mycelium of white colour partially or completely covers the fruit. This manuscript provides a detailed description of the microbes involved in this disease, such as bacteria, yeasts, and moulds. Microbes of different cherry cultivars were studied during two consecutive seasons (2019 and 2020). The counts of bacteria and yeast in damaged tissues were higher (7.05 and 6.38 log10 CFU/g for total aerobic mesophilic microbes and yeasts, respectively) than sound tissues (6.08 and 5.19 log10 CFU/g, respectively). The Enterobacterales order dominated the bacteria population. Among yeasts, Yarrowia lipolytica, in 2019, and Metschnikowia pulcherrima and Metschnikowia viticola, in 2020, were consistently isolated from all samples. The presence of moulds was inconsistently detected at the early stage of this disease by plate counts. However, microscopic observations revealed the presence of hyphae in cherry flesh. Different pathogenic moulds were identified, although white mycelium, identified as Botrytis cinerea by molecular methods, was consistently isolated at later stages. Inoculation tests confirmed the involvement of white-mycelium B. cinerea in the development of this new postharvest disease in the Jerte Valley. Its combination with Enterobacterales enhanced the evolution of rotting, whereas the combination with yeasts decreased and delayed the symptoms. This work presents the first report of a consortia of microorganisms implicated in the development of 'aqueous spot', an emerging disease in sweet cherry cultivars in the Jerte Valley.

Improving the Viability and Metabolism of Intestinal Probiotic Bacteria Using Fibre Obtained from Vegetable By-Products.

This study evaluated the effect of dietary fibre obtained from pomegranate, tomato, grape and broccoli by-products on the gastrointestinal transit survival, growth, and metabolism of six probiotic strains. The results showed that the studied by-products contained variable amounts of polysaccharides that affected the six probiotic microorganisms in different ways. In addition, the protective effect of the fibre obtained on the probiotic strains was more effective in the case of the fibre obtained from tomato peel. In terms of growth, grape stems showed the best results, favouring the growth of lactic acid bacteria. Finally, all fibres were able to increase the content of short-chain fatty acids in the in vitro test, but broccoli stems and pomegranate peel stimulated higher production of short-chain fatty acids. The results of this study demonstrate that plant by-product fibres can improve survival, growth, and metabolism in terms of the fatty acid profiles of probiotic strains, highlighting the desirability of harnessing these by-product fibres to develop new high-value-added ingredients as probiotic carriers.

Chemical Composition and Functional Properties of Dietary Fibre Concentrates from Winemaking By-Products: Skins, Stems and Lees.

The objective of this study was to evaluate, from a technological and nutritional point of view, the chemical composition and functional properties of the industrial winemaking by-products, namely skins, stems and lees. The chemical and physical characteristics, as well as the functional properties (fat and water retention and swelling capacity, antioxidant capacity, and their prebiotic effect), of the dietary fibre of these by-products were studied. The results showed that the skins, stems, and lees are rich in fibre, with the stem fibre containing the highest amounts of non-extractable polyphenols attached to polysaccharides with high antioxidant activity and prebiotic effect. Lee fibre had the highest water retention capacity and oil retention capacity. The results reveal that winemaking by-products could be used as a source of dietary fibre with functional characteristics for food applications.

Strategies to Increase the Biological and Biotechnological Value of Polysaccharides from Agricultural Waste for Application in Healthy Nutrition.

Nowadays, there is a growing interest in the extraction and identification of new high added-value compounds from the agro-food industry that will valorize the great amount of by-products generated. Many of these bioactive compounds have shown beneficial effects for humans in terms of disease prevention, but they are also of great interest in the food industry due to their effect of extending the shelf life of foods by their well-known antioxidant and antimicrobial activity. For this reason, an additional research objective is to establish the best conditions for obtaining these compounds from complex by-product structures without altering their activity or even increasing it. This review highlights recent work on the identification and characterization of bioactive compounds from vegetable by-products, their functional activity, new methodologies for the extraction of bioactive compounds from vegetables, possibly increasing their biological activity, and the future of the global functional food and nutraceuticals market.

Antioxidant, Antihypertensive and Antimicrobial Properties of Phenolic Compounds Obtained from Native Plants by Different Extraction Methods.

This study aims to evaluate the efficacy of two methods (agitation and ultra-sound) for extracting phenolic compounds from 15 native plants. Plant species collected in the Dehesa of Extremadura were used. The antioxidant, antihypertensive and antimicrobial activity of the phenolic extracts was investigated. Significantly different results were obtained when comparing the two extraction methods, with the highest concentrations of phenolic compounds found for ultrasound extraction. In addition, the extracts obtained for Cistus albidus, Cistus salviifolius, Rubus ulmifolius and Quercus ilex showed the highest concentrations of phenolic compounds. The antioxidant activity was higher in the extracts of Cistus and Q. ilex obtained by ultrasound, as was the antihypertensive activity. Antimicrobial activity was also higher in the extracts obtained by ultrasound from C. salviifolius and Q. ilex plants against bacteria and from Cistus ladanifer against yeasts. Therefore, it can be concluded that, with the ultrasound extraction of phenolic compounds from C. ladanifer, C. albidus and Q. ilex plants, it is possible to obtain extracts with important functional properties, so they could be studied for their use in food with the aim of obtaining healthy and safe products, favouring the sustainability of the environment of the Dehesa Extremeña.

Effect of Temperature during Drying and Storage of Dried Figs on Growth, Gene Expression and Aflatoxin Production.

Dried fig is susceptible to infection by Aspergillus flavus, the major producer of the carcinogenic mycotoxins. This fruit may be contaminated by the fungus throughout the entire chain production, especially during natural sun-drying, post-harvest, industrial processing, storage, and fruit retailing. Correct management of such critical stages is necessary to prevent mould growth and mycotoxin accumulation, with temperature being one of the main factors associated with these problems. The effect of different temperatures (5, 16, 25, 30, and 37 °C) related to dried-fig processing on growth, one of the regulatory genes of aflatoxin pathway (aflR) and mycotoxin production by A. flavus, was assessed. Firstly, growth and aflatoxin production of 11 A. flavus strains were checked before selecting two strains (M30 and M144) for in-depth studies. Findings showed that there were enormous differences in aflatoxin amounts and related-gene expression between the two selected strains. Based on the results, mild temperatures, and changes in temperature during drying and storage of dried figs should be avoided. Drying should be conducted at temperatures >30 °C and close to 37 °C, while industry processing, storage, and retailing of dried figs are advisable to perform at refrigeration temperatures (<10 °C) to avoid mycotoxin production.

Anti-fungal activity of phenolic sweet orange peel extract for controlling fungi responsible for post-harvest fruit decay.

There is a growing interest in finding safe and natural anti-microbial compounds as a valid alternative to conventional chemical treatments for managing post-harvest fruit diseases. This study investigated the anti-fungal capacity of orange peel polyphenolic extract (OPE) against three relevant post-harvest fungal pathogens, Monilinia fructicola, Botrytis cinerea and Alternaria alternata. OPE extract at 1.5 g/L inhibited (100%) the mycelial growth and conidial germination of the three target fungi. At lower concentration, the effect varied, depending on the dose applied and target fungi. When the anti-fungal activity of the main phenolic compounds in sweet orange peel, namely, the flavonoids (naringin, hesperidin and neohesperidin) and phenolic acids (ferulic and p-coumaric), were evaluated, ferulic acid and p-coumaric acid displayed significantly higher inhibitory capacity in synthetic medium, while the activity of flavonoids was limited. Synergism between compounds was not detected, and the inhibitory activity of OPE may be attributed to an additive effect of phenolic acids. Interestingly, in peach-based medium, ferulic acid remained active against M. fructicola and A. alternata and was more efficient than p-coumaric to control B. cinerea. These results highlight peel orange waste as an excellent source of anti-fungal compounds, suggesting the possibility of using ferulic acid or ferulic acid-rich extracts, either alone or in combination with other post-harvest treatment, as a natural alternative to reduce post-harvest losses and, also, enhance the shelf-life of fruit.

Cyclopiazonic acid gene expression as strategy to minimizing mycotoxin contamination in cheese.

Expression of genes associated with cyclopiazonic acid (CPA) biosynthesis by Penicillium strains in a cheese-based medium has not been previously studied. To control CPA biosynthesis, it would be useful to understand the changes in gene expression during cheese production and relate them to toxin production. The objective was to evaluate the influence of pH, aw, and temperature on expression of dmaT, which encodes the enzyme dimethylallyl tryptophan synthase involved in the biosynthesis of CPA. We assayed three Penicillium strains, Penicillium commune CBS311 and CBS341 and Penicillium camemberti CBS273, using reverse transcription real-time PCR. Our results showed that the expression patterns of the gene were influenced by strain and environmental conditions. The highest expression for the P. commune strains was observed at pH 6.0, 0.95 aw, at 25 or 30 °C, depending on the strain. In contrast, P. camemberti CBS273 showed a lower dmaT expression with a maximum at 25 °C, pH 5.0 and 0.95 aw. Correlation analysis indicated that the three toxigenic strains showed a strong correlation between the relative expression of the dmaT gene and concentration of CPA under conditions simulating cheese ripening. This method could be used to control CPA production in cheese by detection of dmaT expression.

Evaluation of the Physicochemical and Sensory Characteristics of Different Fig Cultivars for the Fresh Fruit Market.

Physicochemical and sensory properties of nine fig cultivars: 'San Antonio' (SA), 'Blanca Bétera' (BB), 'Brown Turkey' (BT), 'Tres Voltas L'Any' (TV), 'Banane' (BN), 'Cuello Dama Blanco' (CDB), 'Cuello Dama Negro' (CDN), 'Colar Elche' (CE), and 'De Rey' (DR), were compared at three different ripening stages. Weight, size, titratable acidity, pH, skin and flesh colours, firmness, maturation index (MI), and volatile compounds were determined in samples from two consecutive seasons, in addition to both descriptive and hedonic sensory analysis. The mechanical behaviour of figs determined by firmness analysis and colour changes in both skin and flesh was the most important trait for the discrimination of ripening stages. Notable differences among cultivars were found in most of the parameters studied, in particular the inter-cultivar differences highlighted for MI, pH, acidity, and skin colour analyses, followed by volatile compounds. Principal component analysis (PCA) indicated that MI, pH, colour parameters of flesh (h and L*), and terpenes were the best physicochemical indices to determine overall acceptability which is highly correlated with the sensory attributes flesh colour and fruit flavour. The results suggested that CDN and SA showed huge consumer acceptability among the fig cultivars studied.

Effect of Rearing System on the Straight and Branched Fatty Acids of Goat Milk and Meat of Suckling Kids.

Goat meat is considered healthy because it has fewer calories and fat than meat from other traditional meat species. It is also rich in branched chain fatty acids that have health advantages when consumed. We studied the effects of maternal milk and milk replacers fed to suckling kids of four breeds on the straight and branched fatty acid compositions of their muscle. In addition, the proximal and fatty acid compositions of colostrum and milk were studied. Goat colostrum had more protein and fat and less lactose than milk. Goat milk is an important source of healthy fatty acids such as C18:1 c9 and C18:2 n-6. Suckling kid meat was also an important source of C18:1c9. Dairy goat breeds had higher percentages of trans monounsaturated fatty acids (MUFAs) and most of the C18:1 isomers but lower amounts of total MUFAs than meat breeds. However, these dairy kids had meat with a lower percentage of conjugated linoleic acid (CLA) than meat kids. The meat of kids fed natural milk had higher amounts of CLA and branched chain fatty acids (BCFAs) and lower amounts of n-6 fatty acids than kids fed milk replacers. Both milk and meat are a source of linoleic, α-linolenic, docosahexaenoic, eicosapentaenoic and arachidonic fatty acids, which are essential fatty acids and healthy long-chain fatty acids.

Effect of the rearing system on the color of four muscles of suckling kids.

Most suckling kids are raised on farms for cheese production, and many goat farmers rear kids with milk replacers. The aim of this work was to study the influence of the use of milk replacers on the color of four muscles. A total of 246 suckling kids of eight breeds were slaughtered to achieve carcasses of 5 kg. The color of the biceps femoris, semimembranosus, semitendinosus, and longissimus thoracis muscles was measured with a spectrophotometer, and CIELab coordinates were registered. In addition, the pH of longissimus thoracis was measured. The effect of the rearing system (RS) on the color of the studied muscles is strongly modulated by breed. In general terms, there are two groups of kids according to the color of meat. The first group has great lightness and hue angle including Malagueña, Palmera, and Tinerfeña fed natural and artificial milk. The second group with great redness includes Retinta, Payoya, and Verata fed natural and artificial milk together with Florida fed natural milk and Cabra del Guadarrama fed milk replacers. Hence, farms should consider selecting a breed and RS together. Most of the kid meat with high pH comes from kids raised on milk replacers. Because artificial RSs use very early weaning, which might induce a high pH and dark meat, two artificial rearing strategies can be proposed. The first strategy is to choose less sensitive breeds that produce meat with a normal pH. The second strategy is to restrict suckling of natural milk but minimize separation from the mother.

Estudio del estado nutricional y de la prevalencia de exceso de peso en adolescentes de localidades extremeñas de diferentes tamaños / A study of the nutritional status and prevalence of overweight among adolescents from different size towns in Extremadura

RESUMEN En este trabajo se determina el estado nutricional de adolescentes de Extremadura y la prevalencia de sobrepeso y obesidad en función del tamaño de la localidad de procedencia. Se realizó un estudio con 816 estudiantes de Educación Secundaria Obligatoria y Bachillerato, de ambos sexos, con edades de 13 a 18 años. Los centros educativos pertenecían a municipios de una Región española, Extremadura, de diferentes tamaños (de menos de 5.000 a más de 60.000 habitantes). Se recogieron datos antropométricos y, siguiendo criterios nacionales e internacionales, se determinaron los porcentajes de sobrepeso y obesidad en función del tamaño de las localidades. Los alumnos de localidades rurales (menores de 5.000 habitantes) tienden a presentar valores de diferentes parámetros antropométricos que se relacionan con un menor desarrollo corporal. Aunque hubo variaciones en la prevalecía de sobrepeso y obesidad en función de los criterios utilizados, las localidades de más de 60.000 habitantes presentaron los mayores porcentajes medios de exceso de peso y las rurales, los menores.

ABSTRACT The aim of this study was to determine the nutritional status and estimate the prevalence of overweight and obesity among adolescents from different size towns in the Extremadura Region of Spain. A sample of 816 secondary school students of both sexes between 13 and 18 years of age were evaluated. The educative centers selected belonged to different size towns (populations between <5,000 and >60,000 inhabitants). Different anthropometric measures were taken. The percentage of teenagers with overweight and obesity was calculated following national and international standards and compared by town size. Students from rural towns, <5,000 inhabitants, had the lowest values in some of the anthropometric measures; results that are associated with a lower body development. There were differences in the prevalence of overweight and obesity based on the criteria used, but, in general, adolescents from larger towns (>60,000 inhabitants) showed the highest percentages of overweight, whereas rural populations showed the lowest percentages.

Bacterial Communities in Serpa Cheese by Culture Dependent Techniques, 16S rRNA Gene Sequencing and High-throughput Sequencing Analysis.

Serpa cheese is one of the traditional regional Portuguese cheeses having the Protected Denomination of Origin (PDO) designation. This study investigated the bacterial community in the traditional Portuguese Serpa cheese. The microorganisms identified at the end of ripening (30 days) mainly were lactic acid bacteria (LAB). Lactobacillus paracasei/Lactobacillus casei was the main species in cheese from PDO registered industries, whereas in non-PDO registered industries Lactobacillus brevis was highlighted, among other LAB. Enterobacteriaceae species were detected at 20% to 40% of the total isolates. The results obtained by high-throughput sequencing analysis confirmed that LAB was the main microbial group, with Lactococcus genus contributing to approximately 40% to 60% of the population, followed by Leuconostoc and Lactobacillus. The Enterobacteriaceae family was also important. The differences between bacterial communities from PDO and non-PDO registered industries suggest that the lack of regulation of the cheese-making practices may influence unfavorably. The new knowledge about bacterial diversity in Serpa cheese could be useful to set up new ripening conditions, which favor the development of desirable microorganisms. PRACTICAL APPLICATION: The control of the manufacturing process of traditional cheeses can be improved through the knowledge of the bacterial diversity that develops. Thus, the growth of desirable microorganisms can be promoted to homogenize the final product.

Yeast community in traditional Portuguese Serpa cheese by culture-dependent and -independent DNA approaches.

This study investigated the yeast community present in the traditional Portuguese cheese, Serpa, by culture-dependent and -independent methods. Sixteen batches of Serpa cheeses from various regional industries registered with the Protected Designation of Origin (PDO) versus non-PDO registered, during spring and winter, were used. Irrespective of the producer, the yeast counts were around 5log CFU/g in winter and, overall, were lower in spring. The yeast species identified at the end of ripening (30days), using PCR-RFLP analysis and sequencing of the 26S rRNA, mainly corresponded to Debaryomyces hansenii and Kluyveromyces marxianus, with Candida spp. and Pichia spp. present to a lesser extent. The culture-independent results, obtained using high-throughput sequencing analysis, confirmed the prevalence of Debaryomyces spp. and Kluyveromyces spp. but, also, that Galactomyces spp. was relevant for three of the five producers, which indicates its importance during the early stages of the cheese ripening process, considering it was not found among the dominant viable yeast species. In addition, differences between the identified yeast isolated from cheeses obtained from PDO and non-PDO registered industries, showed that the lack of regulation of the cheese-making practice, may unfavourably influence the final yeast microbiota. The new knowledge provided by this study of the yeast diversity in Serpa cheese, could be used to modify the cheese ripening conditions, to favour desirable yeast species. Additionally, the prevalent yeast isolates identified, Debaryomyces hansenii and Kluyveromyces spp., may have an important role during cheese ripening and in the final sensorial characteristics. Thus, the study of their technological and functional properties could be relevant, in the development of an autochthonous starter culture, to ensure final quality and safety of the cheese.