Određivanje aflatoksina, okratoksina A, fumonizina i zearalenona u žitaricama i krmivu primjenom kompetetivnoga direktnog imunoenzimatskog testa (CD-ELISA) i tankoslojne kromatografije (TLC)

Aspergillus, Penicillium, and Fusarium species frequently contaminate crops. For this reason mycotoxins such as afl atoxins (AFs), ochratoxin A (OTA), fumonisins (FBs), and zearalenone (ZEA) are found in food and feed in a wide range of concentrations, depending on environmental and storage conditions. Consumption of mycotoxin-contaminated food and feed has been associated with acute and chronic poisoning and carcinoma. The aim of this study was to determine the incidence and co-occurrence of AFs (B1+B2+G1+G2), OTA, FBs (B1+B2+B3), and ZEA in 37 samples of cereals and feed randomly collected in 2007 from households of an endemic nephropathy (EN) area in Croatia. The mycotoxins were determined using the competitive direct ELISA test (CD-ELISA) in combination with thin-layer chromatography (TLC). The most frequent mycotoxin was ZEA (92 %, mean 318.3 μg kg-1), followed by FBs (27 %, 3690 μg kg-1), AFs (24.3 %, 4.6 μg kg-1), and OTA (16.2 %, 9.8 μg kg-1). Levels of AFs, ZEA, and FBs detected by CD-ELISA signifi cantly correlated with the TLC results. However, only one OTA-positive sample was confi rmed by TLC due to its high limit of detection. The levels of these mycotoxins were below the permissible limit for animal feed. Twenty-nine percent of cereals were contaminated with FBs, OTA, or ZEA in mass fractions above the permissible limit for humans. Co-occurrence of two toxins varied between 4.2 % and 54 % and of three between 4.2 % and 7.6 %. Prolonged co-exposure to AFs, OTA, FBs, and ZEA might increase the risk of various chronic diseases.Vrste plijesni iz rodova Aspergillus, Penicillium i Fusarium česti su kontaminanti usjeva te na takvim supstratima tvore mikotoksine. Stoga su žitarice i krmiva često kontaminirana afl atoksinima (AFs), okratoksinom A (OTA), fumonizinima (FBs) i zearalenonom (ZEA) u različitim koncentracijama ovisno o mikroklimatskim uvjetima na polju i u skladištu. Konzumiranje hrane kontaminirane mikotoksinima često je povezano s akutnim ili kroničnim trovanjima, ali i s razvojem karcinoma. Cilj ovog rada bio je odrediti istodobnu pojavnost AFs (B1+B2+G1+G2), OTA, FBs (B1+B2+B3) i ZEA u uzorcima žitarica i krme (N=37) koji su nasumično skupljeni u individualnim domaćinstvima na području endemske nefropatije (EN) u Hrvatskoj (2007). Za određivanje navedenih mikotoksina korišten je kompetitivni direktni ELISA-test (CD-ELISA) u kombinaciji s tankoslojnom kromatografi jom (TLC). Najzastupljeniji mikotoksin bio je ZEA (92 %, srednja koncentracija 318.3 μg kg-1), nakon čega slijede FBs (27 %, 3690 μg kg-1), AFs (24.3 %, 4.6 μg kg-1) te OTA (16.2 %, 9.8 μg kg-1). Koncentracije AFs, FBs i ZEA određene CD-ELISA-testom statistički značajno koreliraju s rezultatima dobivenim s TLC. OTA je potvrđen metodom TLC samo u jednom uzorku zbog visokog limita detekcije. Dokazane koncentracije su ispod razina dopuštenih za krmiva, dok je 29 % uzoraka žitarica sadržavalo FBs, OTA ili ZEA u koncentracijama iznad dopuštenih u hrani za ljude. Kokontaminacija s dvama odnosno trima toksinima varirala je između 4.2 % i 54 % odnosno između 4.2 % i 7.6 %. Dugotrajni unos AFs, OTA, FBs i ZEA putem hrane može povećati rizik od razvoja različitih kroničnih bolesti zbog njihova mogućega sinergističkog djelovanja

Interakcije nekih plijesni i aflatoksinogenog soja Asspergillus flavus NRRL 3251

The objective of this study was to evaluate biotic interaction between some mould species and active producer of aflatoxin B1 Aspergillus flavus NRRL 3251, co-cultured in yeast-extract sucrose (YES) broth. Twenty-five mould strains of Alternaria spp., Cladosporium spp., Mucor spp., A. flavus and A. niger, used as biocompetitive agents, were isolated from outdoor and indoor airborne fungi, scrapings of mouldy household walls, and from stored and post-harvest maize. Aflatoxin B1 was extracted from mould biomasses with chloroform and detected using the multitoxin TLC method. The results confirm antagonistic interaction between all strains tested. With Alternaria spp. and Cladosporium spp., aflatoxin B1 production decreased 100 %, compared to detection in a single culture of A. flavus NRRL 3251 (Cmean=18.7 µg mL-1). In mixed cultures with Mucor spp., aflatoxin B1 levels dropped to (5.6-9.3) µg mL-1, and the inhibition was from 50 % to 70 %. Four of five aflatoxin non-producing strains of A. flavus interfered with aflatoxin production in mixed culture, and reduced AFB1 productivity by 100 %. One strain showed a lower efficacy in inhibiting AFB1 production (80 %) with a detectable amount of AFB1 3.7 µg mL-1 when compared to control. A decrease in toxin production was also observed in dual cultivation with A. niger strains. It resulted in 100 % reduction in three strains), 90 % reduction in one strain (Cmean=1.9 µg mL-1) and 80 % reduction in one strain (Cmean=3.7 µg mL-1) inhibition.Cilj rada bio je procijeniti biotske interakcije između sojeva različitih vrsta plijesni i kontrolnog soja Aspergillus flavus NRRL 3251, producenta aflatoksina B1 (AFB1). Inhibitorno djelovanje u miješanim kulturama na tvorbu AFB1 ispitano je na dvadeset pet sojeva Alternaria, Cladosporium, Mucor i Aspergillus vrsta izoliranih iz zraka, strugotina pljesnivih zidova te uskladištenog i prezimljenog kukuruza. Biosinteze su provedene u tekućoj hranjivoj podlozi s kvaščevim ekstraktom (YESbujon). Ekstrakcije AFB1 iz biomase izvršene su multitoksinskom metodom tankoslojne kromatografije. Rezultati biotskih interakcija pokazali su antagonistički odnos svih testiranih sojeva. Alternaria i Cladosporium vrste simultano inokulirane sporama A. flavus NRRL 3251 inhibirale su tvorbu AFB1 100 % u odnosu na dokazani toksin u kontrolnoj biosintezi (konc. 18,7 µg mL-1). U miješanim kulturama vrstama roda Mucor dokazane su padajuće koncentracije AFB1 (9,3 µg mL-1, 7,5 µg mL-1 i 5,6 µg mL-1), odnosno inhibicija tvorbe toksina 50 % do 70 %. Atoksinogeni sojevi A. flavus inhibirali su tvorbu AFB1 80 % (1 soj, konc. 3,7 µg mL-1) i 100 % (4 soja). Antagonističko djelovanje prema toksinogenom soju, smanjujući tvorbu AFB1 u rasponu 80 % do 100 % (konc. 1,9 µg mL-1 i 3,7 µg mL-1), dokazano je u uzgojnim biosintezama s A. niger

T-2 toksin - pojavnost i toksičnost u peradi

T-2 toxin is the most toxic type A trichothecene mycotoxin. It is the secondary metabolite of the Fusarium fungi, and is common in grain and animal feed. Toxic effects have been shown both in experimental animals and in livestock. It has been implicated in several outbreaks of human mycotoxicoses. Toxic effects in poultry include inhibition of protein, DNA, and RNA synthesis, cytotoxicity, immunomodulation, cell lesions in the digestive tract, organs and skin, neural disturbances and low performance in poultry production (decreased weight gain, egg production, and hatchability). Concentrations of T-2 toxin in feed are usually low, and its immunosuppressive effects and secondary infections often make diagnosis difficult. If at the onset of the disease, a change in diet leads to health and performance improvements in animals, this may point to mycotoxin poisoning. Regular control of grain and feed samples is a valuable preventive measure, and it is accurate only if representative samples are tested. This article reviews the incidence and toxic effects of T-2 toxin in poultry.T-2 toksin je najtoksičniji predstavnik trikotecenskih mikotoksina tipa A. On je sekundarni produkt metabolizma plijesni roda Fusarium i često je prisutan u žitaricama i hrani za životinje. Štetni učinci uočeni su u eksperimentalnih životinja i životinja u uzgoju. On se povezuje s pojavom bolesti ljudi od mikotoksikoza. Učinci toksina u peradi su višestruki: inhibicija sinteze proteina, DNA i RNA, citotoksični učinak, imunomodulatorni učinak, oštećenje stanica probavnog sustava, organa i kože, živčani poremećaji te pad proizvodnih karakteristika u uzgoju peradi (slabiji prirast, pad nesivosti i valivosti). Koncentracije T-2 toksina u hrani redovito su vrlo malene, a zbog imunosupresivnog djelovanja toksina te istodobne sekundarne infekcije bolest se često teško dijagnosticira. Pri pojavi bolesti promjenom hrane može doći do poboljšanja zdravstvenog stanja, što tako|er upućuje na moguće trovanje mikotoksinima. Redovita kontrola uzoraka žitarica i hrane za životinje jedna je od preventivnih mjera, a detekcija mikotoksina u žitaricama i hrani pouzdana je samo ako se ispituje reprezentativan uzorak. U radu su opisani učestalost i toksični učinci T-2 toksina u peradi

Kontaminacija zrna pšenice, kukuruza, soje i graška vrstama Fusariuma u Hrvatskoj

From 2002 to 2008, 203 samples of wheat, maize, soybean, and pea were analysed for the presence of Fusarium species. Contamination with Fusarium spp., expressed as the percentage of seeds with Fusarium colonies, ranged from 5 % to 69 % for wheat, from 25 % to 100 % for maize, from 4 % to 17 % for soybean, and from 3 % to 17 % for pea. 187 isolates were collected and the following 19 species determined: F. graminearum, F. poae, F. avenaceum, F. verticillioides, F. sporotrichioides, F. heterosporum, F. crookwellense, F. tricinctum, F. semitectum, F. oxysporum, F. proliferatum, F. solani, F. equiseti, F. pseudograminearum, F. chlamydosporum, F. sambucinum, F. compactum, F. scirpi, and F. culmorum. Dominant species were F. graminearum on wheat (27 % of isolates), F. verticillioides on maize (83 % of isolates), F. sporotrichioides on soybean (34 % of isolates), and F. proliferatum on pea (29 % of isolates). Among species identifi ed, F. heterosporum, F. crookwellense, F. pseudograminearum, F. sambucinum, and F. compactum have been reported for the fi rst time in Croatia.U periodu od 2002. do 2008. g. analizirana je prisutnost vrsta Fusariuma na 208 uzoraka zrna pšenice, kukuruza, soje i graška. Kontaminacija vrstama Fusariuma, izražena kao postotak sjemenki s kolonijama Fusarium spp., kretala se od 5 % do 69 % na pšenici, od 25 % do 100 % na kukuruzu, od 4 % do 17 % na soji te od 3 % do 17 % na grašku. Prosječna kontaminacija vrstama Fusariuma u različitim godinama varirala je od 10 % do 46 % na pšenici, od 50 % do 91 % na kukuruzu, od 5 % do 9 % na soji te od 7 % do 10 % na grašku. Vrste Fusariuma koje se javljaju na zrnu izolirane su i determinirane s odabranih uzoraka pšenice, kukuruza, soje i graška. Skupljeno je 187 izolata, a utvrđeno je 19 vrsta: F. graminearum, F. poae, F. avenaceum, F. verticillioides, F. sporotrichioides, F. heterosporum, F. crookwellense, F. tricinctum, F. semitectum, F. oxysporum, F. proliferatum, F. solani, F. equiseti, F. pseudograminearum, F. chlamydosporum, F. sambucinum, F. compactum, F. scirpi i F. culmorum. Dominantne vrste bile su F. graminearum na pšenici (27 % izolata), F. verticillioides na kukuruzu (83 % izolata), F. sporotrichioides na soji (34 % izolata) te F. proliferatum na grašku (29 % izolata). U Hrvatskoj su prvi put utvrđene vrste F. heterosporum, F. crookwellense, F. pseudograminearum, F. sambucinum i F. compactum

The role of open-air inhalatoria in the air quality improvement in spa towns

Objectives: The present study was aimed at evaluating microbiological contamination of air in Ciechocinek and Ino­wro­cław – Polish lowland spa towns. Additionally, the impact of open-air inhalatoria on the quality of air was evaluated. Material and Methods: Air samples were collected seasonally in the urban areas, in the recreation areas and in the vicinity of inhalatoria in both towns using impaction. The numbers of mesophilic bacteria, staphylococci, hemolytic bacteria and actinomycetes were determined on media according to the Polish Standard PN-86/Z-04111/02. The number of moulds was determined on media according to the Polish Standard PN-86/Z-04111/03. Results: While the highest numbers of microorganisms were noted at the sites located in the urban areas, the lowest numbers were noted in the vicinity of the open-air inhalatoria. In all the investigated air samples the values of bioaerosol concentrations were below the recommended TLVs (≤ 5000 CFU×m–3 for both bacteria and fungi in outdoor environments). Location of the sampling site was invariably a decisive factor in determining the number of microorganisms in the air. Conclusions: The aerosol which is formed in the open-air inhalatoria has a positive influence on microbiological air quality. Owing to a unique microclimate and low air contamination, Ciechocinek and Inowrocław comply with all necessary requirements set for health resorts specializing in treating upper respiratory tract infections