Pantić, Vladimir R.

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In vitro degradation of diacetoxyscirpenol and T-2 toxin by use of Mucor racemosus fresen. f. racemosus isolate

Bočarov-Stančić, Aleksandra; Stanković, Slavica; Lević, Jelena; Salma, Nataša M.; Pantić, Vladimir R.; Barnić, Saša S.

(Matica srpska, Novi Sad, 2011)

TY  - JOUR
AU  - Bočarov-Stančić, Aleksandra
AU  - Stanković, Slavica
AU  - Lević, Jelena
AU  - Salma, Nataša M.
AU  - Pantić, Vladimir R.
AU  - Barnić, Saša S.
PY  - 2011
UR  - http://rik.mrizp.rs/handle/123456789/383
AB  - Under controlled in vitro conditions the capacity of the Mucor racemosus f. racemosus 1215/09 isolate to degrade type A trichothecenes (diacetoxyscirpenol - DAS and T-2 toxin) was observed in the liquid nutritive medium. According to previously performed experiments it was proved that the selected isolate, originating from sunflower meal, had the ability to degrade these fusariotoxins when growing on the modified Vogel's agar supplemented with crude extracts of DAS and T-2 toxin. In order to determine biodegradation of fusariotoxins, the liquid nutritive medium - SPY (5% sucrose + 0.1% peptone + 0.1% yeast extract, pH 6.2) was simultaneously inoculated with the isolate M. racemosus f. racemosus 1215/09 and: a) Fusarium semitectum SL-B (DAS producer) or b) F. sporotrichioides R-2301 (T-2 toxin producer). The SPY media, inoculated with single fungal isolates, were used as a control of toxin biosynthesis. The cultures were incubated at room temperature (21-26ºC) on the rotary shaker (175 rpm). After the 3-5-day incubation, the filtration of liquid cultures and the extraction of fusariotoxins from filtrates with ethyl-acetate were performed. Determinations of DAS and T-2 toxin were done by thin layer chromatography using silica gel G. Depending on the incubation duration, M. racemosus f. racemosus in the mixed culture with F. semitectum degraded from 90.0 to 99.97% of DAS present in the medium (40,000- 120,000 µg l-1), while in the mixed culture with F. sporotrichioides it degraded from 95.0 to 96.7% of T-2 toxin present in the medium (240,000 µg l-1). Sterile filtrates of mixed cultures and single culture of M. racemosus f. racemosus, obtained by passing liquid cultures through the 0.45-µm membrane filter and added to the SPY medium, did not affect degradation of type A trichothecenes that had been biosynthesised by isolates F. semitectum SL-B and F. sporotrichioides R-2301 in the liquid medium.
AB  - U kontrolisanim in vitro uslovima proučavana je sposobnost izolata Mucor racemosus f. racemosus 1215/09 da degraduje trihotecene tipa A (diacetoksiscirpenol - DAS i T-2 toksin) u tečnoj hranljivoj podlozi. Prethodnim eksperimentima je dokazano da odabrani izolat, poreklom sa suncokretove sačme, poseduje sposobnost razgradnje navedenih fuzariotoksina, koji su kao sirovi ekstrakti dodati u modifikovanu Vogelovu podlogu. U cilju utvrđivanja biodegradacije fuzariotoksina tečna hranljiva podloga SPK (5% saharoza + 0,1% pepton + 0,1% ekstrakt kvasca, pH 6,2) je zasejana u isto vreme izolatom M. racemosus f. racemosus 1215/09 i: a) Fusarium semitectum SL-B (proizvođač DAS-a) ili b) F. sporotrichioides R-2301 (proizvođač T-2 toksina). Kao kontrola biosinteze toksina korišćena je SPK podloga inokulisana pojedinačnim izolatima gljiva. Kulture su inkubirane na rotacionoj tresilici (175 o/min) tokom 3-5 dana na sobnoj temperaturi (21-26ºC). Nakon 3 do 5 dana inkubacije vršeno je filtriranje tečnih kultura i ekstrakcija fuzariotoksina iz filtrata etil-acetatom. Determinacija DAS-a i T-2 toksina je rađena tankoslojnom hromatografijom na silika gelu G. Zavisno od dužine inkubacije, M. racemosus f. racemosus je u združenoj kulturi sa F. semitectum degradovala 90,0-99,97% DAS-a prisutnog u podlozi (40.000-120.000 µg l-1), dok je u združenoj kulturi sa F. sporotrichioides razgradila 95,0-96,7% T-2 toksina prisutnog u podlozi (240.000 µg l-1). Sterilni filtrati mešanih kultura i pojedinačne kulture M. racemosus f. racemosus, dobijeni propuštanjem tečnih kultura kroz 0,45 µm membranski filter i dodati SPK podlozi, nisu uticali na razgradnju trihotecena tipa A koje su biosintetisali izolati F. semitectum SL-B i F. sporotrichioides R-2301 u tečnoj podlozi.
PB  - Matica srpska, Novi Sad
T2  - Zbornik Matice srpske za prirodne nauke
T1  - In vitro degradation of diacetoxyscirpenol and T-2 toxin by use of Mucor racemosus fresen. f. racemosus isolate
T1  - In vitro degradacija diacetoksiscirpenola i T-2 toksina posredstvom izolata Mucor racemosus fresen. f. racemosus
IS  - 121
SP  - 51
EP  - 59
DO  - 10.2298/ZMSPN1121051B
UR  - conv_58
ER  - 
@article{
author = "Bočarov-Stančić, Aleksandra and Stanković, Slavica and Lević, Jelena and Salma, Nataša M. and Pantić, Vladimir R. and Barnić, Saša S.",
year = "2011",
abstract = "Under controlled in vitro conditions the capacity of the Mucor racemosus f. racemosus 1215/09 isolate to degrade type A trichothecenes (diacetoxyscirpenol - DAS and T-2 toxin) was observed in the liquid nutritive medium. According to previously performed experiments it was proved that the selected isolate, originating from sunflower meal, had the ability to degrade these fusariotoxins when growing on the modified Vogel's agar supplemented with crude extracts of DAS and T-2 toxin. In order to determine biodegradation of fusariotoxins, the liquid nutritive medium - SPY (5% sucrose + 0.1% peptone + 0.1% yeast extract, pH 6.2) was simultaneously inoculated with the isolate M. racemosus f. racemosus 1215/09 and: a) Fusarium semitectum SL-B (DAS producer) or b) F. sporotrichioides R-2301 (T-2 toxin producer). The SPY media, inoculated with single fungal isolates, were used as a control of toxin biosynthesis. The cultures were incubated at room temperature (21-26ºC) on the rotary shaker (175 rpm). After the 3-5-day incubation, the filtration of liquid cultures and the extraction of fusariotoxins from filtrates with ethyl-acetate were performed. Determinations of DAS and T-2 toxin were done by thin layer chromatography using silica gel G. Depending on the incubation duration, M. racemosus f. racemosus in the mixed culture with F. semitectum degraded from 90.0 to 99.97% of DAS present in the medium (40,000- 120,000 µg l-1), while in the mixed culture with F. sporotrichioides it degraded from 95.0 to 96.7% of T-2 toxin present in the medium (240,000 µg l-1). Sterile filtrates of mixed cultures and single culture of M. racemosus f. racemosus, obtained by passing liquid cultures through the 0.45-µm membrane filter and added to the SPY medium, did not affect degradation of type A trichothecenes that had been biosynthesised by isolates F. semitectum SL-B and F. sporotrichioides R-2301 in the liquid medium., U kontrolisanim in vitro uslovima proučavana je sposobnost izolata Mucor racemosus f. racemosus 1215/09 da degraduje trihotecene tipa A (diacetoksiscirpenol - DAS i T-2 toksin) u tečnoj hranljivoj podlozi. Prethodnim eksperimentima je dokazano da odabrani izolat, poreklom sa suncokretove sačme, poseduje sposobnost razgradnje navedenih fuzariotoksina, koji su kao sirovi ekstrakti dodati u modifikovanu Vogelovu podlogu. U cilju utvrđivanja biodegradacije fuzariotoksina tečna hranljiva podloga SPK (5% saharoza + 0,1% pepton + 0,1% ekstrakt kvasca, pH 6,2) je zasejana u isto vreme izolatom M. racemosus f. racemosus 1215/09 i: a) Fusarium semitectum SL-B (proizvođač DAS-a) ili b) F. sporotrichioides R-2301 (proizvođač T-2 toksina). Kao kontrola biosinteze toksina korišćena je SPK podloga inokulisana pojedinačnim izolatima gljiva. Kulture su inkubirane na rotacionoj tresilici (175 o/min) tokom 3-5 dana na sobnoj temperaturi (21-26ºC). Nakon 3 do 5 dana inkubacije vršeno je filtriranje tečnih kultura i ekstrakcija fuzariotoksina iz filtrata etil-acetatom. Determinacija DAS-a i T-2 toksina je rađena tankoslojnom hromatografijom na silika gelu G. Zavisno od dužine inkubacije, M. racemosus f. racemosus je u združenoj kulturi sa F. semitectum degradovala 90,0-99,97% DAS-a prisutnog u podlozi (40.000-120.000 µg l-1), dok je u združenoj kulturi sa F. sporotrichioides razgradila 95,0-96,7% T-2 toksina prisutnog u podlozi (240.000 µg l-1). Sterilni filtrati mešanih kultura i pojedinačne kulture M. racemosus f. racemosus, dobijeni propuštanjem tečnih kultura kroz 0,45 µm membranski filter i dodati SPK podlozi, nisu uticali na razgradnju trihotecena tipa A koje su biosintetisali izolati F. semitectum SL-B i F. sporotrichioides R-2301 u tečnoj podlozi.",
publisher = "Matica srpska, Novi Sad",
journal = "Zbornik Matice srpske za prirodne nauke",
title = "In vitro degradation of diacetoxyscirpenol and T-2 toxin by use of Mucor racemosus fresen. f. racemosus isolate, In vitro degradacija diacetoksiscirpenola i T-2 toksina posredstvom izolata Mucor racemosus fresen. f. racemosus",
number = "121",
pages = "51-59",
doi = "10.2298/ZMSPN1121051B",
url = "conv_58"
}
Bočarov-Stančić, A., Stanković, S., Lević, J., Salma, N. M., Pantić, V. R.,& Barnić, S. S.. (2011). In vitro degradation of diacetoxyscirpenol and T-2 toxin by use of Mucor racemosus fresen. f. racemosus isolate. in Zbornik Matice srpske za prirodne nauke
Matica srpska, Novi Sad.(121), 51-59.
https://doi.org/10.2298/ZMSPN1121051B
conv_58
Bočarov-Stančić A, Stanković S, Lević J, Salma NM, Pantić VR, Barnić SS. In vitro degradation of diacetoxyscirpenol and T-2 toxin by use of Mucor racemosus fresen. f. racemosus isolate. in Zbornik Matice srpske za prirodne nauke. 2011;(121):51-59.
doi:10.2298/ZMSPN1121051B
conv_58 .
Bočarov-Stančić, Aleksandra, Stanković, Slavica, Lević, Jelena, Salma, Nataša M., Pantić, Vladimir R., Barnić, Saša S., "In vitro degradation of diacetoxyscirpenol and T-2 toxin by use of Mucor racemosus fresen. f. racemosus isolate" in Zbornik Matice srpske za prirodne nauke, no. 121 (2011):51-59,
https://doi.org/10.2298/ZMSPN1121051B .,
conv_58 .
2

The role of Paecilomyces lilacinus (Thom) Samson and other fungal species in biodegradation of ochratoxin A

Bočarov-Stančić, Aleksandra; Lević, Jelena; Salma, Nataša M.; Stanković, Slavica; Pantić, Vladimir R.; Dolić, Bisera J.

(Matica srpska, Novi Sad, 2011)

TY  - JOUR
AU  - Bočarov-Stančić, Aleksandra
AU  - Lević, Jelena
AU  - Salma, Nataša M.
AU  - Stanković, Slavica
AU  - Pantić, Vladimir R.
AU  - Dolić, Bisera J.
PY  - 2011
UR  - http://rik.mrizp.rs/handle/123456789/386
AB  - Nine isolates of fungi of genera Aspergillus, Fusarium, Paecilomyces and Penicillium were cultured on the modified Vogel's medium with the addition of crude ochratoxin A (OTA) extract. This crude OTA extract was derived from a natural solid substrate on which Aspergillus ochraceus strain CBS 108.08 was cultivated. OTA was isolated, partially purified, dried by evaporating and dissolved in ethanol (1 mg ml-1), and added to the test medium up to the final concentration of 10 μg ml-1. The presence of OTA residues was determined after 7 and 14 day cultivation of fungi in the test medium at 27±1°C. The Paecilomyces lilacinus isolate (Inf. 2/A), which completely degraded OTA (150 μg) after only seven days, was selected for further studies. Wet sterile rice grains (50 g + 25 ml distilled water) were inoculated with individual isolates of fungi A. ochraceus (strain CBS 108.08) and P. lilacinus (isolate Inf. 2/A), and with their combination. In the case of P. lilacinus monoculture, 0.9 mg of crude OTA was also added into cultivation substrate. Each test was done in three replications. After the four week cultivation of individual and combined fungi at 27±1°C, inoculated rice grains were dried to the constant weight and pulverized. OTA was determined in these samples by the application of standard TLC method for fodder analysis. OTA in the amount of 61.310 μg kg-1 dry matter (DM) was determined only in the samples inoculated with a producer of ochratoxin A (A. ochraceus, strain CBS 108.08). On the other hand, a much smaller amount of OTA (80 μg kg-1 DM) was detected in samples inoculated with combined cultures of A. ochraceus and P. lilacinus isolates. Gained results indicate that P. lilacinus degraded, on average, 99.8% of OTA. After four week cultivation, the same fungal isolate in the samples of wet sterile rice kernels with the addition of 0.9 mg of crude OTA, completely degraded added crude OTA ( lt 8 μg kg-1).
AB  - Devet izolata gljiva iz rodova Aspergillus, Fusarium, Paecilomyces i Penicillium gajeno je na modifikovanoj Vogelovoj podlozi sa dodatkom sirovog ekstrakta ohratoksina A (OTA). Sirovi ekstrakt OTA je dobijen iz čvrstog prirodnog supstrata na kojem je gajen soj Aspergillus ochraceus CBS 108.08. Izolovan i delimično prečišćen OTA, uparen do suvog ostatka i rastvoren u etanolu (1 mg ml-1), dodat je u test podlogu do finalne koncentracije 10 μg ml-1. Nakon sedam i 14 dana gajenja kultura gljiva u test podlozi na 27 ± 1°C de terminisano je prisustvo rezidua OTA primenom modifikovane metode Filtenborg-a i sar. (1983). Od devet testiranih izolata za dalja ispitivanja je odabran izolat Paecilomyces lilacinus (Inf. 2/A), koji je već posle sedam dana u potpunosti razgradio inicijalnu količinu OTA (150 μg). U drugom delu eksperimenta vlažno sterilno zrno pirinča (50 g + 25 ml destilovane vode) zasejano je sa pojedinačnim izolatima A. ochraceus (CBS 108.08) i P. lilacinus (Inf. 2-A), kao i kombinacijom oba izolata. U slučaju mono-kulture P. lilacinus u podlogu je dodat i sirovi OTA (0,9 mg). Svaki od testova je urađen u 3 ponavljanja. Nakon četiri nedelje gajenja monokultura i mešanih kul tura gljiva na 27±1°C, inokulisana zrna su osušena do konstantne težine i sa mlevena do finog praha. U ovim uzorcima izvršena je determinacija OTA primenom standardne metode tankoslojne hromatografije za analizu stočne hrane. U uzorcima koji su bili zasejani samo sa producentom OTA (A. ochraceus, soj CBS 108.08) detektovan je OTA u prosečnoj količini od 61.310 μg kg-1 suvog ostatka. U uzorcima koji su bili zasejani kombinovanim kulturama izolata A. ochraceus i P. lilacinus utvrđena je znatno manja prosečna količina OTA (80 μg kg-1). Ovi rezultati ukazuju da je izolat P. lilacinus razgradio prosečno 99,8% OTA prisutnog u podlozi za kultivaciju. U uzorcima vlažnog sterilnog zrna pirinča sa dodatkom 0,9 mg sirovog OTA isti gljivični izolat je posle četiri nedelje kultivacije kompletno biorazgradio dodat sirovi OTA ( lt 8 μg kg-1).
PB  - Matica srpska, Novi Sad
T2  - Zbornik Matice srpske za prirodne nauke
T1  - The role of Paecilomyces lilacinus (Thom) Samson and other fungal species in biodegradation of ochratoxin A
T1  - Uloga Paecilomyces lilacinus (Thom) Samson i drugih vrsta gljiva u biodegradaciji ohratoksina A
IS  - 120
SP  - 103
EP  - 110
DO  - 10.2298/ZMSPN1120103B
UR  - conv_57
ER  - 
@article{
author = "Bočarov-Stančić, Aleksandra and Lević, Jelena and Salma, Nataša M. and Stanković, Slavica and Pantić, Vladimir R. and Dolić, Bisera J.",
year = "2011",
abstract = "Nine isolates of fungi of genera Aspergillus, Fusarium, Paecilomyces and Penicillium were cultured on the modified Vogel's medium with the addition of crude ochratoxin A (OTA) extract. This crude OTA extract was derived from a natural solid substrate on which Aspergillus ochraceus strain CBS 108.08 was cultivated. OTA was isolated, partially purified, dried by evaporating and dissolved in ethanol (1 mg ml-1), and added to the test medium up to the final concentration of 10 μg ml-1. The presence of OTA residues was determined after 7 and 14 day cultivation of fungi in the test medium at 27±1°C. The Paecilomyces lilacinus isolate (Inf. 2/A), which completely degraded OTA (150 μg) after only seven days, was selected for further studies. Wet sterile rice grains (50 g + 25 ml distilled water) were inoculated with individual isolates of fungi A. ochraceus (strain CBS 108.08) and P. lilacinus (isolate Inf. 2/A), and with their combination. In the case of P. lilacinus monoculture, 0.9 mg of crude OTA was also added into cultivation substrate. Each test was done in three replications. After the four week cultivation of individual and combined fungi at 27±1°C, inoculated rice grains were dried to the constant weight and pulverized. OTA was determined in these samples by the application of standard TLC method for fodder analysis. OTA in the amount of 61.310 μg kg-1 dry matter (DM) was determined only in the samples inoculated with a producer of ochratoxin A (A. ochraceus, strain CBS 108.08). On the other hand, a much smaller amount of OTA (80 μg kg-1 DM) was detected in samples inoculated with combined cultures of A. ochraceus and P. lilacinus isolates. Gained results indicate that P. lilacinus degraded, on average, 99.8% of OTA. After four week cultivation, the same fungal isolate in the samples of wet sterile rice kernels with the addition of 0.9 mg of crude OTA, completely degraded added crude OTA ( lt 8 μg kg-1)., Devet izolata gljiva iz rodova Aspergillus, Fusarium, Paecilomyces i Penicillium gajeno je na modifikovanoj Vogelovoj podlozi sa dodatkom sirovog ekstrakta ohratoksina A (OTA). Sirovi ekstrakt OTA je dobijen iz čvrstog prirodnog supstrata na kojem je gajen soj Aspergillus ochraceus CBS 108.08. Izolovan i delimično prečišćen OTA, uparen do suvog ostatka i rastvoren u etanolu (1 mg ml-1), dodat je u test podlogu do finalne koncentracije 10 μg ml-1. Nakon sedam i 14 dana gajenja kultura gljiva u test podlozi na 27 ± 1°C de terminisano je prisustvo rezidua OTA primenom modifikovane metode Filtenborg-a i sar. (1983). Od devet testiranih izolata za dalja ispitivanja je odabran izolat Paecilomyces lilacinus (Inf. 2/A), koji je već posle sedam dana u potpunosti razgradio inicijalnu količinu OTA (150 μg). U drugom delu eksperimenta vlažno sterilno zrno pirinča (50 g + 25 ml destilovane vode) zasejano je sa pojedinačnim izolatima A. ochraceus (CBS 108.08) i P. lilacinus (Inf. 2-A), kao i kombinacijom oba izolata. U slučaju mono-kulture P. lilacinus u podlogu je dodat i sirovi OTA (0,9 mg). Svaki od testova je urađen u 3 ponavljanja. Nakon četiri nedelje gajenja monokultura i mešanih kul tura gljiva na 27±1°C, inokulisana zrna su osušena do konstantne težine i sa mlevena do finog praha. U ovim uzorcima izvršena je determinacija OTA primenom standardne metode tankoslojne hromatografije za analizu stočne hrane. U uzorcima koji su bili zasejani samo sa producentom OTA (A. ochraceus, soj CBS 108.08) detektovan je OTA u prosečnoj količini od 61.310 μg kg-1 suvog ostatka. U uzorcima koji su bili zasejani kombinovanim kulturama izolata A. ochraceus i P. lilacinus utvrđena je znatno manja prosečna količina OTA (80 μg kg-1). Ovi rezultati ukazuju da je izolat P. lilacinus razgradio prosečno 99,8% OTA prisutnog u podlozi za kultivaciju. U uzorcima vlažnog sterilnog zrna pirinča sa dodatkom 0,9 mg sirovog OTA isti gljivični izolat je posle četiri nedelje kultivacije kompletno biorazgradio dodat sirovi OTA ( lt 8 μg kg-1).",
publisher = "Matica srpska, Novi Sad",
journal = "Zbornik Matice srpske za prirodne nauke",
title = "The role of Paecilomyces lilacinus (Thom) Samson and other fungal species in biodegradation of ochratoxin A, Uloga Paecilomyces lilacinus (Thom) Samson i drugih vrsta gljiva u biodegradaciji ohratoksina A",
number = "120",
pages = "103-110",
doi = "10.2298/ZMSPN1120103B",
url = "conv_57"
}
Bočarov-Stančić, A., Lević, J., Salma, N. M., Stanković, S., Pantić, V. R.,& Dolić, B. J.. (2011). The role of Paecilomyces lilacinus (Thom) Samson and other fungal species in biodegradation of ochratoxin A. in Zbornik Matice srpske za prirodne nauke
Matica srpska, Novi Sad.(120), 103-110.
https://doi.org/10.2298/ZMSPN1120103B
conv_57
Bočarov-Stančić A, Lević J, Salma NM, Stanković S, Pantić VR, Dolić BJ. The role of Paecilomyces lilacinus (Thom) Samson and other fungal species in biodegradation of ochratoxin A. in Zbornik Matice srpske za prirodne nauke. 2011;(120):103-110.
doi:10.2298/ZMSPN1120103B
conv_57 .
Bočarov-Stančić, Aleksandra, Lević, Jelena, Salma, Nataša M., Stanković, Slavica, Pantić, Vladimir R., Dolić, Bisera J., "The role of Paecilomyces lilacinus (Thom) Samson and other fungal species in biodegradation of ochratoxin A" in Zbornik Matice srpske za prirodne nauke, no. 120 (2011):103-110,
https://doi.org/10.2298/ZMSPN1120103B .,
conv_57 .
1