Determination of Zearalenone and their metabolites in sheep urine by ELISA Method.

PDF
Published: Apr 20, 2024
DOI: https://doi.org/10.12681/jhvms.34009
Keywords:
mycotoxins zearalenone zeranol sheep urine ELISA
S Teqja
Laboratory of Toxicology and Veterinary Drug Residues, Department of Toxicology, Food Safety and Veterinary Institute Tirana, Albania
J Mavromati
Department of Veterinary Public Health, Faculty of Veterinary Medicine, Agricultural University of Tirana, Albania
Abstract

The Fusarium metabolite mycotoxin zearalenone (ZEA) is of concern because of its lifelong estrogenic effects in animals. Zeranol as its metabolite was a widely used in animals as growth promoters to increase body weight. But, the use of zeranol as a growth promotion in animals that produce products for human consumption has been banned in the European Union (EU). As no data exist on the occurrence of zearalenone and zeranol in sheep urine in Albania we have analyzed samples of them by ELISA method. The method is based on a competitive enzyme immunoassay, ELISA for the quantitative analysis of zearalenone and its metabolites in urine. The overall recoveries of the analysis were in range of 80% -103%. Eighty nine (n=89) sheep urine samples were taken in the study during the last 2 years until now. Eighty (n= 80) samples were found to be compliant below the Screening Target Concentration (Cut-Off) value (0.404 - 0.839 ng/ml). Nine (n=9) samples were found to be suspect with the ELISA method above the Cut-Off value. After analyzing the suspect samples with the UPLC-MS/MS confirmatory method, two (n=2) sheep urine samples were noncompliant above decision limit for confirmation (CCα). In the first sample concentration found are 1 and 1.5 in the second sample 3 and 4.5 µg/kg for α-Zeralanol and β-Zeralanol respectively. The high levels of α-Zeralanol and β-Zeralanol concentrations found in urine indicate a contamination of animal feed with Zearalenone. This comes as a consequence of poor animal feed storage and leads to the accumulation of ZEA before harvest time or abuse has occurred. Further studies should be conducted to contribute to the knowledge disposition of zearalenone and zeranol in animals to give an accurate answer about possible contamination or illegal use.


 

Article Details
  • Section
  • Research Articles
Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Authors who publish with this journal agree to the following terms:

· Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution Non-Commercial License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.

· Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g. post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.

· Authors are permitted and encouraged to post their work online (preferably in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.

Downloads
Download data is not yet available.
References
Ali N, Degen G H, (2019). Biomonitoring of zearalenone and its main metabolites in urines of Bangladeshi adults. Food and Chemical Toxicology.130: 276–283.
Arts CJM, van Baak MJ, van de Velde-Fase K, Witkamp RF, Elliot CJ, Armstrong HS, Cooper J (1998). Comparison of conventional immunoassays and the oestrogen radioreceptor assay for screening for the presence of oestrogenic anabolic compounds in urine samples. The Analyst. 123(12): 2579-2583.
Bagnati R, Oriundi M P, Russo V, Danese M, Berti F, Fanelli R (1991). Determination of zeranol and β-zearalanol in calf urine by immunoaffinity extraction and gas chromatography—mass spectrometry after repeated administration of zeranol. Journal of ChromatographyB: Biomedical Sciences and Applications. 564(2): 493–502.
Bennekom EO, Brouwer L, Laurant EHM, Hooijerink H, Nielen MWF (2002). Confirmatory analysis method for zeranol, its metabolites and related mycotoxins in urine by liquid chromatography-negative ion electrospray tandem mass spectrometry.AnalyticaChimicaActa. 473(1-2): 151–160.
’’COMMUNITY REFERENCE LABORATORIES RESIDUES (CRLs).”https://food.ec.europa.eu/system/files/2016-10/cs_vet-med-residues_guideline_validation_screening_en.pdf[accessed 20 January 2010].
Commission Decision 2002/657/EC - BVLhttps://eurl-residues.eu/wp-content/uploads/2020/12/EURL_MMPR_guidance-paper_final.pdf[accessed September 2020]. Commission Implementing Regulation (EU) 2021/808 of 22 https://eur-lex.
europa.eu/legal-content/EN/TXT/PDF/?uri=CELEX:32021R0808&-from=EN[accessed 22 March 2021].
Council Directive 96/22/EChttps://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?ri=CELEX:31996L0022&from=EN[accessed 29 April 1996].
Cooper KM, Ribeiro L, Alves P, Vozikis V, Tsitsamis S, Alfredsson G, Lövgren T, Tuomola M, Takalo H, Iitia A, Sterk SS, Blokland M, Kennedy DG. (2003). Interlaboratory ring test of time-resolved fluoroimmunoassays for zeranol and α-zearalenol and comparison with zeranol test kits. Food Additives and Contaminants. 20(9):804-812.
Cooper K M, Tuomola M, Lahdenperä S, Lövgren T, Elliott C T, Kennedy DG. (2002). Development and validation of dry reagent time-resolved fluoroimmunoassays for zeranol and α -zearalenol to assist in distinguishing zeranol abuse from Fusarium spp. toxin contamination in bovine urine. Food Additives and Contaminants. 19(12):1130-1137.
Dänicke S, Winkler J, (2015). Invited review: Diagnosis of zearalenone (ZEN) exposure of farm animals and transfer of its residues into edible tissues (carry over). Food and Chemical Toxicology. 84: 225-249.
EURL GUIDANCE ON MINIMUM METHOD PERFORMANCE REQUIREMENTS (MMPRs) FOR SPECIFIC PHARMACOLOGICALLY
ACTIVE SUBSTANCES IN SPECIFIC ANIMAL MATRICEShttps://eurl-residues.eu/wp-content/uploads/2020/12/EURL_MMPR_guidance-paper_final.pdf[accessed September 2020].
Jana N, Nanda S, (2018). Resorcylic acid lactones (RALs) and their structural congeners: recent advances in their biosynthesis, chemical synthesis and biology. New Journal of Chemistry.
Krout-Greenberg ND, Puschner B, Davidson MG, DePeters EJ. (2013). Preliminary study to assess mycotoxin oncentrations in whole corn in the California feed supply. Journal of Dairy Science. 96(4):2705-2712.
Knutsen H‐K, Alexander J, Barregård L,Bignami M, Brüschweiler B, Ceccatelli S, Cottrill B, Dinovi M, Edler L, Grasl‐Kraupp B, Hogstrand C, Hoogenboom LR, Nebbia CS, Petersen A, Rose M, Roudot AC, Schwerdtle T, Vleminckx C, Vollmer G, Wallace H, Dall’AstaC, Dänicke S, Eriksen GS, Altieri A, Roldán‐Torres R, Oswald IP (2017). Risks for animal health related to the presence of zearalenone and its modified forms in feed. EFSA Journal, 15(7).
Lafayette, The Presence, Effect, and Diagnosis of Zearalenone in Dairy Cattle West. “Winter 2003 Newsletter.”West Lafayette:406 S. University West Lafayette, IN 47907.
Launay FM, Ribeiro L, Alves P, Vozikis V, Tsitsamis S, Alfredsson G, Sterk SS, Blokland M, Iitia A, Lövgren T, Tuomola M, Gordo, Kennedy DG (2004). Prevalence of zeranol, taleranol and Fusarium spp. toxins in urine: implications for the control of zeranol abuse in the European Union. Food Additives and Contaminants. 21(9): 833-839.
Launay FM, Ribeiro L, Alves P, Vozikis V, Tsitsamis S, Alfredsson G, Sterk SS, Blokland M, Iitia A, Lövgren T, Tuomola M, Gordon A, Kennedy DG (2004). Prevalence of zeranol, taleranol and Fusarium spp. toxins in urine: implications for the control of zeranol abuse in the European Union. Food Additives and Contaminants. 21(9):833-839
Launay FM, Young PB, Sterk SS, Blokland MH, Kennedy DG (2004) Confirmatory assay for zeranol, taleranol and the Fusarium spp. toxins in bovine urine using liquid chromatography-tandem mass spectrometry. Food Additives and Contaminants. 21(1):52-62.
Liu Y, Zhang C, Yu X, Zhang Zh, Zhang X, Liu R, Liu X, Gong Zh (2007) Development and evaluation of immunoassay for zeranol in bovine urine. Journal of Zhejiang University SCIENCE B. 8(12): 900-905.
Miles CO, Erasmuson, A F, Wilkins AL, Towers NR, Smith B L, Garthwaite I, Scahill G, Bryan G, Hansen RP (1996). Ovine Metabolism of Zearalenone to α-Zearalanol (Zeranol). Journal of Agricultural and Food Chemistry. 44(10): 3244-3250.
Rai A, Das M, Tripathi A (2019). Occurrence and toxicity of a fusarium mycotoxin, zearalenone. Critical Reviews in Food Science and Nutrition. 1-20.
Scientific Opinion on the risks for public health related to the presence of zearalenone in food. (2011) EFSA Journal, 9(6): 2197.
Takagi M, Uno S, Kokushi E, Shiga S, Mukai S, Kuriyagawa T, Takagaki K, Hasunuma H, Matsumoto D, Okamoto K, Shahada F, Chenga T, Deguchi E, Fink-Gremmels J (2011). Measurement of urinary zearalenone concentrations for monitoring natural feed contamination in cattle herds: On-farm trials. Journal of Animal Science. 89(1): 287-296.
Wang S, Wang XH (2007). Analytical methods for the determination of zeranol residues in animal products. Food Additives and Contaminants. 24(6): 573–582.
Yang Ch, Song G, Lim W (2020). Effects of Mycotoxin-Contaminated Feed on Farm Animals. Journal of Hazardous Materials. 122087.
Yin M, Hu Xi, Sun Y, Xing Y, Chai Sh, Xing G, Yang Y, Teng M, Li Q, Wang Y, Deng R, Zhang G (2020). The broad-spectrum and ultra-sensitive detection of zeranol and its analogues by an enzyme-linked immunosorbent
assay in cattle origin samples. RSC Advances. 10(35):20809-20816.
Liu Y, Zhang C, Yu X, Zhang Z, Zhang X, Liu R, Liu X,, Gong Zh (2007). Development and evaluation of immunoassay for zeranol in bovine urine. Journal of Zhejiang University SCIENCE B. 8(12):900-905.
Zhang G-L, Feng Y-L, Song J-L, Zhou X-Sh (2018). Zearalenone: A Mycotoxin With Different Toxic Effect in Domestic and Laboratory Animals’ Granulosa Cells. Frontiers in Genetics. Volume
Most read articles by the same author(s)