Uticaj Min-a-zela Plus na biohemijske, mikrobiološke i mikotoksikološke parametre silaža lucerke

Lucerne of the 5th cut in the budding phase was ensiled in the experiment as fresh and wilted (DM= 220 and 360 g/kg), with two degrees of compression (520 and 380 g/dm3) and with added Min-a-zel plus as 2 g/kg green mass. The experiment was performed as the statistical model 2K. Min-a-zel plus used as 2 g/kg green mass had no influence on biochemical, microbiological and mycotoxicological parameters of lucerne silage. Compared to the beginning material, silages had more diverse microflora and more mycotoxins. All silages had high pH values and fermentation of the acetic acid type. Some improvement was achieved with wilting of the ensiling material. The least quality (V class) was obtained with fresh lucerne, which was less compressed, while others were ranked as IV quality class. The obtained results fortify the need for further investigations in developing correct doses of Min-a-zel plus to be used in ensiling of lucerne and other materials.U eksperimentu je silirana lucerka V otkosa u fazi butonizacije kao sveža i provenula (SM = 220 i 360 g/kg), sa dva stepena sabijenosti (520 i 380 g/dm3) i sa dodatkom Min-a-zela Plus, 2 g/kg zelene mase. Ogled je postavljen po statističkom modelu 2K. Min-a-zel Plus korišćen u dozi od 2 g/kg zelene mase nije uticao na biohemijske, mikrobiološke i mikotoksikološke parametre silaža lucerke. U odnosu na početni materijal silaže su sadržale raznovrsniju mikrofloru i veću količinu mikotoksina. Sve silaže su se odlikovale visokim pH vrednostima i fermentacijom sirćetnokiselinskog tipa. Izvesno poboljšanje je postignuto provenjavanjem biljnog materijala. Najlošiji kvalitet (V klasa) imale su silaže od sveže lucerke sa manjim stepenom sabijenosti, dok su ostale ocenjene IV klasom kvaliteta. Utvrđeni rezultati ukazuju na neophodnost nastavljanja ispitivanja u cilju utvrđivanja optimalne doze Min-a-zela Plus za siliranje lucerke i drugih sirovina

Uticaj mineralnog adsorbenta 'Minazel' na smanjenje rezidua zearalenona i njegovih metabolita u mleku krava

The results of the use of mineral adsorbent - Minazel, preparation based on the natural zeolite, in the prevention of mycotoxicoses and avoidance of zearalenone and its metabolites transfer into cow´s milk were described in this paper. Using mineral adsorbent Minazel at the concentration of 0,5% of concentrate mixture the transfer of zearalenone from feeds to milk was prevented.U radu su prikazani rezultati korišćenja adsorbenta mikotoksina - Minazel preparata na bazi prirodnog zeolita, u prevenciji mikotoksikoza i sprečavanju prelaska (akumuliranja) zearalenona i njegovih metabolita u mleku krava. Mikotoksikološkim analizama mleka utvrđeno je da je prisustvo zearalenona u mleku (0,053; 0,019 i 0,004 mg/kg) opadalo sa povećanjem količine Minazela u koncentratu (0; 0,2; 0,5), što uz ostvareno povećanje proizvodnje mleka opravdava njegovo korišćenje

Uticaj mineralnog adsorbenta 'Minazel” na smanjenje rezidua zearalenona i njegovih metabolita u mleku krava

The results of the use of mineral adsorbent - Minazel, a preparation based on natural zeolite, in the prevention of mycotoxicoses and avoidance of zearalenone and its metabolites transfer into cow's milk were described in this paper. Using mineral adsorbent Minazel in a concentration of 0.5% of concentrate mixture the transfer of zearalenone from feeds to milk was prevented.U radu su prikazani rezultati korišćenja adsorbenta mikotoksina - Minazel, preparata na bazi prirodnog zeolita, u prevenciji mikotoksikoza i sprečavanju prelaska (akumuliranja) zearalenona i njegovih metabolita u mleko krava. Mikotoksikološkim analizama mleka utvrđeno je da je prisustvo zearalenona u mleku (0,053; 0,019 i 0,004 mg/kg) opadalo sa povećanjem količine Minazela u koncentratu (0; 0,2; 0,5), što uz ostvareno povećanje proizvodnje mleka opravdava njegovo korišćenje

Učestalost toksigenih vrsta Fusarium i fuzariotoksina u zrnu pšenice u Srbiji

A total of 88 and 40 wheat samples collected immediately prior to harvest in 2005 and 2006, respectively, under different agroecological conditions, were studied in respect to the occurrence of Fusarium spp. and the production of fusariotoxins. The greatest number of samples was infected with species of the genera Fusarium (81.8 and 65.0%), and Alternaria (36.3 and 17.5%) with the intensity ranging from 9.4 to 84.0% in 2005 and from 23.4 to 80.6% in 2006. Out of 13 identified species belonging to the genus Fusarium, F. graminearum had the highest frequency (35.2 and 12.5%) and the intensity up to 67.2%, and 21.9%, in 2005 and 2006, respectively, followed by F. poae but only in 2005 (20.4%), and F. proliferatum in 2006 (19.7%). The natural occurrence of mycotoxins in positive samples varied from 37 to 331 ppb for zearalenone and from 31 to 125 ppb for diacetoxyscirpenol (DAS) and T-2 toxin. The concentration of mycotoxins amounted, on average, to 133.4, 61.0 and 45.7 ppb for zearelenone, DAS and T-2 toxin, respectively.Osamdeset osam i 40 uzoraka pšenice, prikupljenih neposredno pred žetvu u 2005. i 2006. godini u različitim agroekološkim uslovima u Srbiji proučavani su radi praćenja pojave Fusarium spp. i stvaranja fuzariotoksina. Najveći broj uzoraka bio je zaražen vrstama roda Fusarium (81,8 i 65,0%), a zatim sa Alternaria spp. (36,3 i 17,5%) sa intenzitetom 9,4-84,0% u 2005. i 23,4-80,6% u 2006. godini. Od ukupno 13 identifikovanih vrsta iz roda Fusarium najučestalija je bila F. graminearum (35,2 i 12,5%) sa intenzitetom do 67,2% u 2005. i do 21,9% u 2006. godini, a zatim F. poae, ali samo u 2005. godini (20,4%), i F. proliferatum u 2006. godini (19.7%). Prirodna pojava mikotoksina u pozitivnim uzorcima je varirala od 37 do 331 ppb za zearalenon i od 31 do 125 ppb za diacetoksiscirpenol (DAS) i T-2 toksin. U proseku, koncentracija mikotoksina je bila 133,4 ppb za zearelonon, 61,0 ppb za DAS i 45,7 ppb za T-2 toksin

Toksigeni potencijal izolata Fusarium poae poreklom sa pšenice

Eleven isolates of F. poae, originated from wheat grain at 9 locations mainly in Vojvodina, were encompassed by the present study. The greatest number of samples was collected in 2005, in which the climatic conditions favoured a more intensive occurrence of Fusarium ear blight of wheat. In order to determine toxicological potential of this species, cultures of the selected isolates were grown in liquid media (GPY and SPY) on a rotary shaker (180 revolutions min-1), at room temperature (21-26°C) for three days. Crude toxins were isolated from liquid culture filtrates of isolates by the use of ethyl acetate, while quantification of mycotoxins was done by the thin layer chromatography method. A liquid culture of the isolate GZ-LES (F. graminearum) was used as a control for the evaluation of the zearalenone biosynthesis potential. On the other hand, the liquid culture of the isolate KF-38/1 (F. sporotrichioides) was used as a control for both type-A trichothecenes (T-2 toxin and diacetoxyscirpenol - DAS). The obtained results show that F. poae, in contrast to F. graminearum, has no potential for the zearalenone biosynthesis. The presence of DAS was determined only in one isolate of F. poae (MRIZP-666), and in the control isolate of F. sporotrichioides (KF-38/1/R), that were grown in the GPY liquid medium. The T-2 toxin was detected in the isolate MRIZP-666, grown in both media, and in the isolates MRIZP-37 and MRIZP-860, cultured in the GPY and SPY liquid medium, respectively. The control culture KF-38/1/R (F. sporotrichioides) produced the T-2 toxin at the concentration of 4,000 μg L-1. According to the gained information, it can be concluded that the potential of F. poae for the type-A trichothecene biosynthesis was low, as the concentration of DAS or T-2 toxin did not exceed 80 μg L-1 or 240 μg L-1, respectively.U ovom radu je u in vitro uslovima proučena sposobnost izolata F. poae za biosintezu jedne grupe fuzariotoksina - trihotecena tipa A (T-2 toksin i diacetoksiscirpenol - DAS), kao i zearalenona (ZEA). Toksikološki profil ove vrste je kod nas nedovoljno ispitan s obzirom na njenu zastupljenost i toksigena svojstva prema literaturnim podacima. Proučavanjima je bilo obuhvaćeno 11 izolata F. poae, poreklom sa pšenice iz 9 lokaliteta, uglavnom sa područja Vojvodine. Najveći broj uzoraka prikupljen je 2005. godine, kada su klimatski uslovi pogodovali intenzivnijoj pojavi fuzarioza klasa ove poljoprivredne kulture. Za određivanje toksikološkog potencijala F. poae kulture odabranih izolata su gajene u tečnim podlogama (GPK i SPK) tokom 3 dana na sobnoj temperaturi (21-26°C) i na rotacionoj tresilici (180 obrtaja min-1). Sirovi toksini su izolovani iz filtrata tečnih kultura ispitanih izolata pomoću etil acetata, dok je kvantifikacija mikotoksina izvršena metodom tankoslojne hromatografije. Tečna kultura izolata GZ-LES (F. graminearum) je korišćena kao kontrolna kultura za utvrđivanje potencijala za biosintezu zearalenona, a KF-38/1/R (F. sporotrichioides) za oba trihotecena tipa A (T-2 toksin i DAS). Dobijeni rezultati pokazuju da F. poae, za razliku od F. graminearum, ne poseduje potencijal za biosintezu zearalenona. Prisustvo DAS-a je utvrđeno samo kod jedne kulture F. poae (MRIZP-666) i kontrolnog izolata F. sporotrichioides (KF-38/1/R) koji su gajeni u tečnom GPK medijumu. T-2 toksin je detektovan kod izolata MRIZP-666 pri gajenju u obema podlogama, kao i izolata MRIZP-37 u GPK, odnosno MRIZP-860 u SPK medijumu. Kontrolna kultura KF-38/1/R (F. sporotrichioides) proizvodila je T-2 toksin u koncentraciji od 4000 μg L-1. Na osnovu iznetih podataka može se zaključiti da je potencijal F. poae za biosintezu trihotecena tipa A bio nizak u datim uslovima s obzirom da koncentracija DAS-a nije prelazila 80 μg L-1, odnosno T-2 toksina 240 μg L-1. Imajući u vidu prikazane rezultate, smatramo da je za dobijanje konačnog odgovora na pitanje o toksikološkom profilu izolata F. poae u Srbiji neophodno preduzeti dodatna ispitivanja, ne samo sa novim izolatima iz godina koje slede, nego i u drugim uslovima kultivisanja, u prvom redu na sterilnom prirodnom supstratu kao što su pšenica i kukuruz

In vitro degradacija diacetoksiscirpenola i T-2 toksina posredstvom izolata Mucor racemosus fresen. f. racemosus

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

Mikopopulacije silaže od lucerke, s posebnim osvrtom na prisustvo toksigenih Fusarium vrsta

Mycological and mycotoxicological investigations of alfalfa samples (initial not fermented plant material, as well as silage obtained from unfaded and faded state of the same) were performed during the year 2003. Total of 14 fungal species, included in 11 genera, were identified during the present study. The most frequent moulds were Chaetomium piluliferum and their anamorph Botryotrichum piluliferum (83.3% and 66.7% respectively). Potentially toxigenic Fusarium spp. (F. culmorum, F. semitectum and F. sporotrichioides) were observed less frequently (from 16.7% to 33.0%) and only on initial not fermented alfalfa. From the sample of alfalfa faded state contaminated with 0.25 mg/kg of diacetoxyscirpenol (DAS) and 1.28 mg/kg of zearalenone (ZEA) F. semitectum SL-B was isolated. The production of fusariotoxins by this strain was later on tested in vitro conditions. Different aeration treatments in semisynthetic medium with glucose (GPK) or sucrose (SPK), as well as cultivation on sterilized corn kernels (moisture content 47%) were used. The highest yield of DAS (64.0 mg/l) was obtained during submerged cultivation of F. semitectum SL-B in GPK (210 rpms, 3 days room temperature). Production of T-2 toxin, but rather poor (0.08 mg/1), was detected only in SPK (150 rpms, 3 days, room temperature). ZEA was found exclusively after 25 days of cultivation on corn kernels at room temperature (21-25°C).Tokom 2003. god. izvršena su mikološka i mikotoksikološka istraživanja uzoraka lucerke (neprovenuli, provenuli polazni materijal i silaža od istih). Identifikovano je ukupno 14 vrsta, odnosno 11 rodova gljiva od kojih su sa najvećom učestalošću zabeleženi Chaetomium piluliferum i njegov anamorf Bolryotrichum piluliferum (83.3 odnosno 66.7%). Potencijalno toksigene Fusarium spp. (F. culmorum, F. semilectum i F. sporotrichioides) uočene su znatno ređe (od 16.7 do 33.0%). Iz uzorka provenule lucerke kontaminirane sa zearalenonom (ZEA) i diacetoksiscirpenolom (DAS) izolovan je soj F. semitectum SL-B čija je sposobnost za biosintezu fuzariotoksina ispitana u in vitro uslovima. Najveći prinos DAS-a (64.0 mg/l) je postignut pri subverznoj kultivaciji u tečnoj podlozi sa glukozom. Slaba proizvodnja T-2 toksina (0.08 mg/kg) je dobijena samo u tečnoj podlozi sa saharozom. ZEA je nađen isključivo pri kultivaciji na vlažnom sterilnom zrnu kukuruza (6.4 mg/kg)

Investigations of the capability of Fusarium isolates from corn for biosynthesis of fusariotoxins

The aim of this paper was to investigate the potential of zearalenone (ZEN) and type A trichothecenes (T-2 toxin and diacetoxyscirpenole - DAS) for biosynthesis by Fusarium spp. isolated from corn kernels contaminated by mycotoxins. The samples of corn kernels (2004 and 2005 harvest) originating in different regions of Bačka (Vojvodina, Serbia) were tested. Mycotoxicological investigations showed in most cases a significant contamination with deoxynivalenol (DON 200 - 2,460 μg/kg) and ZEA (520 1,680 μg/kg). Isolations and identifications of fusaria established only the presence of species F. verticillioides, after one month storage in freezer conditions, in fusariotoxin positive samples. The control cultures known as ZEA producers - F. graminearum GZ-LES, i.e. T-2 toxin and DAS producer - F. sporotrichioides KF-38/1/R were also tested. In vitro toxicological investigations of isolated fusaria were performed in liquid semisynthetic media (GPK or SPK), and on wet sterilized corn kernels, respectively. Under testing conditions, analyzed F. verticillioide and F. sporotrishioides isolates were not ZEA producers. Contrary to them, F. graminearum GZ-LES pure culture was very good producer of fusariotoxins; it biosynthesized max. 465,900 μg/kg DON, and 4,416 μg/kg ZEA, respectively. Cultivation conditions influenced a great deal of T-2 toxin production under laboratory conditions. In most cases, higher yields were obtained during the cultivation of F. verticillioides in liquid glucose medium (80-240 μg/L). Contrary to the control strain F. sporotrichioides KF-38/1/R that under the same conditions synthesized, besides T-2 toxin (4.000 μg/L) and DAS (240 μg/L), isolates of F. verticillioides from corn grain did not show that ability

Mycopopulations of alfalfa silage with particular review on toxigenic Fusarium spp.

Mycological and mycotoxicological investigations of alfalfa samples (initial not fermented plant material, as well as silage obtained from unfaded and faded state of the same) were performed during the year 2003. Total of 14 fungal species, included in 11 genera, were identified during the present study. The most frequent moulds were Chaetomium piluliferum and their anamorph Botryotrichum piluliferum (83.3% and 66.7% respectively). Potentially toxigenic Fusarium spp. (F. culmorum, F. semitectum and F. sporotrichioides) were observed less frequently (from 16.7% to 33.0%) and only on initial not fermented alfalfa. From the sample of alfalfa faded state contaminated with 0.25 mg/kg of diacetoxyscirpenol (DAS) and 1.28 mg/kg of zearalenone (ZEA) F. semitectum SL-B was isolated. The production of fusariotoxins by this strain was later on tested in vitro conditions. Different aeration treatments in semisynthetic medium with glucose (GPK) or sucrose (SPK), as well as cultivation on sterilized corn kernels (moisture content 47%) were used. The highest yield of DAS (64.0 mg/l) was obtained during submerged cultivation of F. semitectum SL-B in GPK (210 rpms, 3 days room temperature). Production of T-2 toxin, but rather poor (0.08 mg/1), was detected only in SPK (150 rpms, 3 days, room temperature). ZEA was found exclusively after 25 days of cultivation on corn kernels at room temperature (21-25°C)

Mikotoksini u silaži - uzroci stvaranja, posledice i zaštita od delovanja

The causes of appearance of mold in the silage, genus and species of fungi which biosynthesize mycotoxins, acting consequences of micotoxins, prevention of fungi growth and possibilities to prevent their negative effects are shown in this paper. Also, the results of mycotoxins presence in the silage (corn and lucerne) in Vojvodina in the period 2000-2004 are presented. The most commonly found mycotoxins were zearale-none in 60.6% of analyzed samples and DAS in 30.3% of samples. Silage contamination with ochratoxin A, aflatoxin B1 and T-2 toxin was between 15.2 and 21.2%. The content of mycotoxins DAS and T2 toxin was above the values allowed by regulative. The solutions which contribute to the prevention of development of the molds and elimination of negative effects of mycotoxins in silage were analyzed and suggested.U radu se ukazuje na uzroke pojave plesni u silaži, rodove/vrste plesni koje biosintetišu mikotoksine, posledice njihovog delovanja, mere sprečavanja razvoja i mogućnosti otklanjanja štetnog uticaja. Istaknuto je da usled propusta u tehnologiji siliranja (nedovoljno gaženje, prisustvo vazduha) pri vlažnosti mase za siliranje, postoje povoljni uslovi za razvoj plesni koje biosintetišu toksine štetne po zdravlje životinja i ljudi. Oni mogu da izazovu zdravstvene poremećaje uključujući i patohistološke promene na organima i tkivima deponuju se u proizvodima (mleku, mesu i jajima) i na taj način ugroze zdravlje ljudi. U radu su izloženi rezultati prisustva mikotoksina u silaži (kukuruz i lucerka) u Vojvodini u periodu 2000-2004. Najčešće prisutan mikotoksin bio je zearalenon u 60,6% analiziranih uzoraka, a potom DAS u 30,3% uzoraka. Kontaminiranost silaže ohratoksinom A, aflatoksinom B1 i T-2 toksinom bila je između 15,2 i 21,2%. Sadržaj mikotoksina DAS i T-2 toksina bio je na granici ili iznad vrednosti koje dozvoljava Pravilnik (Službeni list SRJ, br. 2/90). Vrednosti za ostale mikotoksine bile su ispod maksimalno dozvoljenih količina. Analizirana su i predložena rešenja koja doprinose suzbijanju razvoja plesni i mikotoksina