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Keywords: naravoslovje, raziskovanje, dijaki
Published in RUNG: 09.11.2023; Views: 2192; Downloads: 0
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Abstract: The presence of bioactive compounds in food and beverages of plant origin is mainly connected with higher nutritional value and better sensory properties. However, some of them can pose a threat to food product quality and human health. For example, excess biogenic amines (BAs) intake can cause different allergenic responses in individuals who have such intolerances. BAs have been intensively studied in recent years, especially in fermented foods like wine, meat, fish, and cheese. Among fermented beverages is apple cider still rather unexplored from this perspective. Especially since no such data exist for the Norwegian cider. Norwegian cider is becoming more and more popular in Norway in recent years among producers and consumers. Hardanger cider from Western Norway is very different from French, English, or Spanish ciders in terms of sensory characteristics, apple cultivars, and in the fermentation process. In Hardanger, the traditional cider is still produced by spontaneous fermentation of apple juice with naturally occurring yeasts that originate from the fruit or processing equipment surfaces. Lactic acid bacteria are known to be associated with BA formation. However, several studies reported about the BA-producing yeasts in winemaking. Due to the important role of natural yeasts in the production of Hardanger cider, we focused on the ability of BA formation by native yeasts. Thus, in our study, we followed the amounts of BAs in the Hardanger ciders during the fermentation process and characterize isolated yeasts if they have the ability to produce BAs under cidermaking conditions by performing a micro-fermentation experiment. From must/cider samples, taken during the fermentation process at 13 producers in the Hardanger region, we isolated 530 yeast isolates. Based on the sequencing of the D1/D2 domain of the 26S rDNA we identified 25 different yeast species. As expected, yeast diversity was higher at the beginning compared to the middle fermentation stage, when mostly different non-Saccharomyces yeast species prevailed, while at the end of fermentation mainly Saccharomyces species with high ethanol tolerance were present. BAs were analyzed with the HPLC-UV method. In all apple juice/cider samples 4 different BAs (putrescine, cadaverine, histamine, and tyramine) were detected and quantified with external calibration. On average in all cider samples from the producers total BA (summation of all BAs) concentration reached 9,45 mg/L, however in one case even 25 mg/L. Tyramine was the most abundant BA in all fermentation stages. 40 isolated yeast strains were further tested for BA formation in a small-scale experiment by fermenting apple juice. Results show that non-Saccharomyces yeasts mainly form histamine (1,68 mg/L) and tyramine (1,30 mg/L), while Saccharomyces yeasts putrescine (0,48 mg/L) and tyramine (3,53 mg/L). As a general conclusion, the occurrence and distribution of BA concentrations in the small-scale fermentation were lower (2,96 mg/L and 4,01 mg/L) and less variable than in the real ciders (average in final ciders 9,45 mg/L) and with tyramine being the most abundant BA in all samples analyzed.
Keywords: cider, yeast, biogenic amines, Hardanger, biodiversity
Published in RUNG: 23.06.2023; Views: 1903; Downloads: 7
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Abstract: In the Hardanger area in Western Norway, the production of cider has a long tradition that goes back to the 12th century, when monks introduced apple growing in this area. Nowadays, this is also the main area of fruit production in Norway. Despite the strict regulation of the alcoholic beverage production in Norway, traditional cider is still produced on some farms in this area. Therefore, our aim was to study the ecology and biodiversity of the yeasts associated with the cider production in the Hardanger area during fermentation process; especially of traditional cider, which is produced by a spontaneous fermentation of apple juice, performed by naturally occurring indigenous yeasts that originate from the fruit or the surfaces of the processing equipment. In our study, samples of fermenting juice/cider were taken during fermentation process from 12 producers, located in 12 different locations in Hardanger region. Classical cultivation methods using WL (Wallerstein Laboratories) agar medium with added chloramphenicol enable us to isolate a total of 530 yeast isolates that were stored in in-house yeast collection at the NIBIO and included also at the Wine Research Centre collection. Based on the sequencing of the D1/D2 domain of the 26S rDNA we managed to identify 357 isolates and distinguished 27 different yeast species as follows: Aureobasidium pullulans, Candida californica, C. oleophila, C, sake, Hanseniaspora meyeri, H. uvarum, H. valbyensis. Kregervanrija fluxuum, Kregervanrija sp., Metschnikowia andauensis, M. chrysoperlae, M. fructicola, M. pulcherrima, Metschnikowia sp, Pichia fermentans, P. kluyveri, P. membranifaciens, P. nakasei, Piskurozyma capsuligena, Rhodotorula nothofagi, Saccharomyces bayanus, S. cerevisiae, S. paradoxus, S. pastorianus, Saccharomyces sp., S. uvarum and Torulaspora delbrueckii. Even though we were not able to obtain samples in three different fermentation stages (beginning, middle and at the end of fermentation) from all producers, we could observe yeast succession during fermentation progress. Yeast diversity was higher at the beginning comparing to the middle of fermentation, when mostly different non-Saccharomyces yeast species prevailed, while in the middle of fermentation 11 species were detected (Candida californica, H. uvarum, H. valbyensis, Kregervanrija sp., K. fluxuum, Pichia membranifaciens, Metschnikowia pulcherrima, Saccharomyces sp, S. bayanus, S. uvarum and S. cerevisie). On the other hand, at the end of fermentation mainly Saccharomyces species with high ethanol tolerance were present (Saccharomyces sp., S. cerevisiae, bayanus, S. uvarum and P. fermentans). In samples that were collected from three producers in all three fermentation stages also quality parameters were determined (ethanol, organic acids, sugars, biogenic amines) with in-house developed methods using HPLC-UV/RID. The most important sugars in ciders were fructose and glucose, as expected. Two producers added sugar to increase the level of ethanol in the middle of fermentation, which is a common procedure in the Hardanger area. Ethanol and organic acid analysis indicated that fermentations went in the right direction, since all parameters were within normal limits. Including the acetic acid level, an indicator of low cider quality, was very low (average around 0,06 g/L). The alcohol incised from the beginning to end fermentation in all samples analysed and minimum concentration was 2,71 g/L. In ciders we detected four biogenic amines (putrescin, cadaverine, histamine and tyramine). The average amount was 32 mg/L and the most abundant was tyramine.
Keywords: indigenous yeasts, biodiversity, spontaneous fermentation, cider-making
Published in RUNG: 18.10.2022; Views: 2400; Downloads: 0
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Abstract: Opravili smo dve hitri molekularni tehniki za identifikacijo enoloških kvasovk, ki so bile izolirane iz vinogradov z ekološko in intergrirano pridelavo grozdja. Najprej smo določili 15 Saccharomyces kvasovk s pomočjo analize multipleks PCR med zbranimi 131 izolati, nato smo s pomočjo ribotipizacije ITS določili približno 98 kvasovk do nivoja vrste, ki so bile potrjene tudi s sekvenciranjem regije ITS. Določili smo 16 vrst vinskih kvasovk: Hanseniaspora uvarum, H. valbyensis, H. opuntiae / H. uvarum, Lachancea thermotolerans, Candida tropicalis, Candida sp. / Starmerella sp., Metschnikowia pulcherrima, Pichia kudriavzevii, P. terricola, P. kluyveri, Wickerhamomyces anomalus, Kodamaea ohmeri, S. bacillaris, Saccharomyces cerevisiae, Saturnispora diversa in Torulaspora delbrueckii. Ugotovili smo, da so najdene kvasovke običajne naseljevalke grozdne mikroflore ter da je način pridelave grozdja vplival na pojavljanje določenih vrst v obravnavanih vinogradih. Nadalje smo testirali kvasovke na tvorbo sideroforov, tvorbo protiglivnih hlapnih organskih spojin (HOS), hidrolitično aktivnost in toleranco kvasovk na fungicide, kot so baker, iprodion, fluazinam ter kombinacijo ciprodinil / fludioksonil. Za kvalitativno detekcijo hidrolitične aktivnosti smo uporabili presejalne teste na osnovi trdnega gojišča z dodanim hitinom ali β-D-glukozidi kot substratom in ugotovili, da je veliko testiranih kvasovk sposobnih proizvajati litične encime, ki bi lahko bili odgovorni za razgradnjo celičnih sten fitopatogenih gliv in potencialno sposobni proizvajati HOS s hidrolizo grozdnih glikozidov zaradi prisotnosti β-glukozidaz. Prav tako smo opazili, da so te encimske aktivnosti običajno odvisne od seva. Na splošno so kvasovke tudi tolerantne na testirane fungicide. Ugotovili smo tudi, da je odpornost biofungicidnih kvasovk na fungicide koristna lastnost za razvoj biofungicidov v prihodnosti pri aplikacijah v strategijah z zmanjšanim vnosom pesticidov.
Keywords: Kvasovka, biološki nadzor, fitopatogen, Botrytis cinerea, vinogradništvo
Published in RUNG: 01.12.2019; Views: 5481; Downloads: 134
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Keywords: biotska raznovrstnost, kvasovke, biokontrolna aktivnost, biofungicid, Botrytis cinerea, trajnostno vinogradništvo
Published in RUNG: 28.09.2017; Views: 6238; Downloads: 0
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