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Uticaj ekstrakata žačina na rast plesni i biosintezu mikotoksina / The effect of spice extracts on the growth of moulds and mycotoxinbiosynthesis

<p>Cilj istraživanja doktorske disertacije bio je da se ispita pojedinačni i sinergistički uticaj<br />ekstrakata kima (Carum carvi L.), bosiljka (Ocimum basilicum L.), origana (Origanum<br />vulgare L.) i etarskih ulja crnog (Allium cepa L. kultivar Kupusinski jabučar) i belog luka<br />(Allium sativum L. kultivar Bosut) na rast plesni izolovanih iz prehrambenih proizvoda, kao i<br />njihov uticaj na biosintezu mikotoksina.<br />Ukupan broj plesni u uzorcima salata od povrća spremnih za konzumiranje kretao se od<br />10,0 do 5,5&acute;102 cfu/g, u uzorcima poslastičarskih proizvoda do 6,1&acute;102 cfu/g i u<br />proizvodima od mesa do 60,0 cfu/g. Najveći broj plesni izolovan je na DG18 podlozi<br />(1,53&acute;102 cfu/g), a najmanji na MY50G (42,0 cfu/g). U ukupnoj mikopopulaciji svih ispitivanih<br />uzoraka dominirale su vrste rodova Penicillium (39,07%), Cladosporium (23,40%) i<br />Aspergillus (20,42%). Vrste iz rodova Alternaria, Fusarium i Eurotium su bile zastupljene sa<br />5,85%, 4,97% i 2,76%. Dominantne vrste u ukupnoj mikopopulaciji bile su C.<br />cladosporioides (21,63%), A. niger (16,0%) i P. aurantiogriseum (11,81%).<br />Dominirali su potencijalni producenti ohratoksina A (31,89%), proizvo&ntilde;ači fumonizina<br />(4,74%), moniliformina (1,43%) i sterigmatocistina (1,54%). Izolati A. versicolor su<br />biosintetisali sterigmatocistin u koncentracijama od 56,3 i 109,2 ng/mL. Ostale potencijalne<br />toksin-produkujuće vrste nisu pokazale sposobnost produkcije mikotoksina.</p><p>Mikotoksikolo&scaron;kim ispitivanjem hrane u dva uzorka salata spremnih za konzumiranje<br />(kupus beli rezani i FIT salata) utvr&ntilde;en je sadržaj sterigmatocistina u koncentracijama od 3,5<br />i 5,5 mg/kg.<br />Kao glavna komponenta u ekstraktu kima odre&ntilde;en je karvon (43,98%), u ekstraktu bosiljka<br />estragol (metil kavikol) (86,72%), a u ekstraktu origana karvakrol (34,20%) i karvon (18,05%).<br />Najveći deo etarskog ulja crnog luka činili su: dimetil-trisulfid, metil-propil-trisulfid, dimetiltetrasulfid,<br />dietil-1,2,4-tritiolan, metil-(1-propenil)-trisulfid, metil-(1-propenil)-disulfid. Dialildisulfid,<br />dialil-trisulfid, metil-alil-trisulfid i metil-alil-disulfid su glavne komponente koje su<br />odre&ntilde;ene u etarskom ulju belog luka.<br />Koncentracija od 0,35 mL/100 mL ekstrakta kima je bila fungicidna (MFC) prema C.<br />cladosporioides, dok je 0,70 mL/100 mL potpuno inhibirala rast A. carbonarius, A. wentii, E.<br />nidulans, Eurotium spp., C. cladosporioides, P. glabrum, P. brevicompactum, F.<br />subglutinans i F. verticillioides. Na rast P. chrysogenum i P. aurantiogriseum ista<br />koncentracija bila je inhibitorna (MIC). Najslabije delovanje ovaj ekstrakt ispoljio prema A.<br />niger, A. versicolor, F. oxysporum i F. proliferatum.<br />Primena ekstrakta bosiljka u koncentraciji od 0,70 mL/100 mL pokazala je fungicidno<br />delovanje na C. cladosporioides. Koncentracija od 1,50 mL/100 mL potpuno je inhibirala<br />rast A. wentii, A. versicolor, E. nidulans, E. herbariorum, E. chevalieri, E. rubrum, P.<br />chrysogenum i Fusarim spp. Ekstrakt bosiljka je najslabije delovao prema A. niger, A.<br />carbonarius, P. aurantiogriseum, E. amstelodami, P. glabrum i P. brevicompactum.<br />Ekstrakt origana je pokazao najslabije ihibitorno delovanje na rast ispitivanih plesni.<br />Primena ekstrakta u koncentraciji od 1,50 mL/100 mL je bila fungicidna prema E. rubrum.<br />Koncentracija od 2,50 mL/100 mL je pokazala fungicidno delovanje na E. rubrum, E.<br />herbariorum, A. wentii, C. cladosporioides i P. aurantiogriseum, a inhibitorno prema E.<br />nidulans, E. chevalieri, E. amstelodami, P. glabrum i P. brevicompactum. Ovaj ekstrakt je<br />najslabije delovao na A. niger, A. carbonarius, F. proliferatum, F. subglutinans i P.<br />chrysogenum.<br />Etarsko ulje crnog luka pokazalo je signifikantno jače antifungalno delovanje na ispitivane<br />plesni u odnosu na etarsko ulje belog luka. Koncentracija od 14,0 mL/100 mL ulja belog luka<br />fungicidno je delovala prema E. rubrum, E. chevalieri i C. cladosporioides, dok je ulje crnog<br />luka na ovoj koncentraciji pokazalo fungicidni efekat i na E. herbariorum i E. amstelodami.<br />Za ostale plesni fungicidna koncentracija iznosila je 28,0 mL/100 mL, osim za A. niger i P.<br />aurantiogriseum.<br />Neke od ispitivanih sme&scaron;a bosiljka i kima, bosiljka i origana, origana i kima i etarskih ulja<br />lukova pokazale su sinergističko delovanje na inhibiciju rasta A. wentii, E. herbariorum, F.<br />verticilllioides i P. aurantiogriseum sa FICindex od 0,63 do 0,97.<br />Začinski ekstrakti i etarska ulja lukova su pored ograničavanja rasta kolonija plesni<br />uzrokovali i promene u makro i mikromorfologiji.<br />Potpuna inhibicija biosinteze sterigmatocistina i rasta A. versicolor postignuta je pri<br />koncentraciji od 0,20 mL/100 mL ekstrakta kima i origana u periodu od 21 dana. Na ovoj<br />koncentraciji ekstrakt bosiljka je inhibirao biosintezu sterigmatocistina za 88,73% i rast<br />plesni za 52,56%. Sme&scaron;a koja je sadržavala 75% ekstrakta kima i 25% ekstrakta bosiljka<br />potpuno je inhibirala rast plesni i biosintezu sterigmatocistina u YES bujonu tokom 21 dana<br />inkubiranja.<br />Pojedinačne koncentracije etarskih ulja crnog i belog luka od 5,0 i 10,0 mL/100 mL i u sme&scaron;i<br />sa 1,50 mL/100 mL etarskog ulja crnog luka i 0,50 mL/100 mL etarskog ulja belog luka bile su<br />potrebne za potpunu inhibiciju rasta A. versicolor i biosintezu sterigmatocistina.<br />Dodatak sme&scaron;e ekstrakata kima i bosiljka (0,35 mL/100 mL ekstrakta kima + 0,70 mL/100 mL<br />ekstrakta bosiljka) u svež kupus rezanac uticao je na smanjenje inicijalne kontaminacije<br />plesnima za 93,9%, uz pojavu intenzivnijeg, ali prihvatljivog mirisa i neznatne promene boje.</p><p>Definisani matematički model za komparaciju uticaja ekstrakata i etarskih ulja na rast plesni<br />može se primenjivati u formiranju matrica inhibicije i optimizaciji vremena i koncentracije<br />antifungalnih agenasa.<br />Dobijena saznanja o antifungalnom delovanju ekstrakata začina i etarskih ulja lukova mogu<br />biti značajna u pobolj&scaron;anju antifungalne za&scaron;tite namirnica, smanjenju biosinteze<br />mikotoksina i ukupnim smanjenju &scaron;teta izazvanih delovanjem plesni.</p> / <p>The aim of this PhD thesis was to study the individual and synergistic effects of extracts of<br />caraway (Carum carvi L.), basil (Ocimum basilicum L.), oregano (Origanum vulgare L.) and the<br />essential oils of onion (Allium cepa L. cultivar Kupusinski jabučar) and garlic (Allium sativum<br />L. cultivar Bosut) on the growth of moulds isolated from food products. The study also<br />focused on the impact of extracts and the essential oils on mycotoxins biosynthesis.<br />The total number of moulds detected in samples of vegetable salads &ldquo;ready for use&rdquo; ranged<br />from 10.0 to 5.5&acute;102 cfu/g. In cake and pastries, as well as, meat samples, the number<br />reached 6.1&acute;102 cfu/g and 60.0 cfu/g, respectively. The highest number of mould colonies was<br />isolated in DG18 medium (1.53 &acute; 102cfu/g) and the lowest in MY50G medium (42.0 cfu/g). The<br />species of the genera Penicillium (39.07%), Cladosporium (23.40%) and Aspergillus (20.42%)<br />prevailed in the entire mycopopulation of all tested samples. Species of the genera Alternaria,<br />Fusarium and Eurotium were represented with 5.85%, 4.97% and 2.76%, respectively, while C.<br />cladosporioides (21.63%), A. niger (16.0%) and P. aurantiogriseum (11.81%) were the most<br />dominant species in the entire mycopopulation.<br />Potential producers of ochratoxin A (31.89%) accounted for the largest share of the isolated<br />mycopopulation. The share of producers of fumonisin, moniliformin and sterigmatocystin<br />amounted to 4.74%, 1.43% and 1.54%, respectively. Sterigmatocystin was biosynthesised in</p><p>the concentration of 56.3 ng/mL and 109.2 ng/mL by both isolates of A. versicolor, while other<br />potential toxin producers did not show the ability of mycotoxin production.<br />Mycotoxicological investigation showed the sterigmatocystin content in two samples of<br />vegetable salads &quot;ready for use&quot; (shredded white cabbage and FIT salad - carrot, lettuce and<br />red chicory) in concentrations of 3.5 mg/kg and 5.5 mg/kg, respectively.<br />The major component in the extract of caraway was carvon with a share of 43.98%. The basil<br />extract contained estragol (methyl cavicol) in the highest percentage (86.72%), while<br />carvacrol (34.20%) and carvon (18.05%) were major components of the oregano extract. As for<br />the essential oil of onion, dimethyl trisulphide, methyl propyl trisulphide, dimethyl tetrasulfid,<br />diethyl-1, 2, 4-tritiolan, methyl-(1-propenyl)-trisulphide, and methyl-(1-propenyl) &ndash; disulfide<br />constituted the largest share. The major components isolated in garlic essential oil were<br />diallyl disulfide, diallyl-trisulphide, allyl methyl trisulphide and allyl methyl disulfide.<br />The concentration of the caraway extract of 0.35 mL/100mL exhibited fungicidal effect (MFC)<br />on C. cladosporioides, while the concentration of 0.70 mL/100mL completely inhibited the<br />growth of A. carbonarius, A. wentii, E. nidulans, Eurotium spp., C. cladosporioides, P.<br />glabrum, P. brevicopmactum, F. subglutinans and F. verticillioides. The same concentration<br />showed the inhibitory effect (MIC) on the growth of P. chrysogenum and P. aurantiogriseum.<br />The poorest effect of the caraway extract was expressed on the growth of A. niger, A.<br />versicolor, F. oxysporum and F. proliferatum.<br />The basil extract application in the concentration of 0.70 mL/100mL showed fungicidal effects<br />(MFC) on the growth of C. cladosporioides. The concentration of 1.50 mL/100mL completely<br />inhibited (MFC) the growth of A. wentii, A. versicolor, E. nidulans, E. herbariorum, E.<br />chevalierii, E. rubrum, P. chrysogenum and Fusarim spp. The poorest effect of the basil<br />extract was exhibited on A. niger, A. carbonarius, P. aurantiogriseum, E. amstelodami, P.<br />glabrum and P. brevicompactum.<br />The oregano extract showed the weakest growth inhibition influence on all of the tested<br />moulds. The application of this extract in the concentration of 1.50 mL/100mL was fungicidal<br />(MFC) to E. rubrum. The concentration of 2.50 mL/100mL showed fungicidal effects (MFC) on<br />the growth of E. rubrum, E. herbariorum, A. wentii, C. cladosporioides and P. aurantiogriseum<br />and inhibitory effects (MIC) on E. nidulans, E. chevalieri, E. amstelodami, P. glabrum and P.<br />brevicompactum. The weakest effect of this extract was expressed on the growth of A. niger,<br />A. carbonarius, F. proliferatum, F. subglutinans and P. chrysogenum.<br />Onion essential oil showed a significantly stronger antifungal effect on the tested moulds in<br />comparison to garlic essential oil. While the concentration of 14.0 mL/100mL of garlic oil had a<br />fungicidal effect on E. rubrum, E. chevalieri and C. cladosporioides, the same concentration<br />of onion oil was also fungicidal to E. herbariorum and E. amstelodami. With an exception of A.<br />niger and P. aurantiogriseum, the concentration that showed a fungicidal effect on the<br />remaining moulds equalled 28.0 mL/100mL.<br />Some of the tested mixtures of basil with caraway, basil with oregano, oregano with caraway,<br />and essential oils of onion and garlic, showed a synergistic effect on the growth inhibition of<br />A. wentii, E. herbariorum, F. verticilllioides and P. aurantiogriseum with the FIC index ranging<br />from 0.63 to 0.97.<br />Apart from the inhibitory effect on the mould colony growth, the spices extracts and the<br />essential oils of onion and garlic also caused changes in the macro- and micro- morphology<br />of the moulds.<br />Complete inhibition of the growth of A. versicolor and sterigmatocystin biosynthesis was<br />achieved at a concentration of 0.20 mL/100mL of the extract of caraway and oregano in the<br />period of 21 days. At this concentration the basil extract delayed the sterigmatocystin<br />biosynthesis by 88.73% while the mould growth was inhibited by 52.56%. Mixtures containing</p><p>75% of the caraway extract and 25% of the basil extract completely inhibited the mould<br />growth and sterigmatocystin biosynthesis in YES broth during 21 days of incubation.<br />The concentrations of 5.0 m L/100mL (onion essential oil) and 10.0 m L/100mL (garlic essential<br />oil) applied in a mixture containing 1.50 mL/100mL of onion and 0.50 mL/100mL of garlic<br />essential oil were necessary for a complete inhibition of the growth of A. versicolor and<br />sterigmatocystin biosynthesis.<br />The addition of the mixture of caraway and basil extracts (0.35 mL/100mL of caraway + 0.70<br />mL/100mL of basil) to fresh shredded cabbage influenced the reduction of initial mould<br />contamination by 93.9%. This was accompanied by the occurrence of acceptable more<br />intense flavour and slight discoloration.<br />The defined mathematical model for comparing the effects of extracts and essential oils on<br />the growth of moulds can be applied in establishing inhibition matrices and optimisation of<br />the time and the concentration of antifungal agents.<br />The obtained results on the antifungal effects of the spices extracts and onion and garlic<br />essential oils can be beneficial for improving the antifungal protection of food and reducing<br />the mycotoxin biosynthesis as well as the overall damage caused by the action of moulds.</p>

Identiferoai:union.ndltd.org:uns.ac.rs/oai:CRISUNS:(BISIS)82710
Date18 July 2012
CreatorsKocić-Tanackov Sunčica
ContributorsDimić Gordana, Lević Jelena, Škrinjar Marija, Vukojević Jelena, Pavlović Hrvoje
PublisherUniverzitet u Novom Sadu, Tehnološki fakultet Novi Sad, University of Novi Sad, Faculty of Technology at Novi Sad
Source SetsUniversity of Novi Sad
LanguageSerbian
Detected LanguageUnknown
TypePhD thesis
Formatapplication/pdf

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