Veterynarna biotehnologija – Veterinary biotechnology, 2021, 39, 44-52 [in Ukrainian]. https://doi.org/10.31073/vet_biotech39-04
Institute of Veterinary Medicine NAAS
USE OF POLIMERASE CHAIN REACTION FOR DETECTION OF TOXINPRODUCING FUNGI OF FUSARIUM GENUS
Introduction. The mycotoxicosis issues due to the wide area of disseminationof toxinogenic fungi is relevant because of the spread of poisoning animals with fusariotoxins. According to the published data, 80% of cereals are contaminated with fusarium toxins. As a result of long-term monitoring of micotoxins containmination of seed, food and grain, it was found that 60% of them were containminated, and 12% contained fusariotoxins.
The goal of the work was to detect of toxin-producing species of the genus Fusarium isolated in Ukraine by polymerase chain reaction (PCR).
Materials and methods. It was used 24 isolates of Fusarium genus fungi from museum of the Institute of veterinary medicine NAAS. Studies of morphological and cultural properties were performed using conventional microbiological methods.
The fusariotoxin produsing isolates were determined by multiplex PCR test (conventional method with electrophoretic reading of results) using three sets of primers recommended by Bluhm et al. (2004). The assay parameters were optimized by us.
The results of experimental studies are processed by conventional methods of statistics.
Results of research and discussion. According to the analysis of the results of microbiological studies, it was found that all the studied isolates had the main typical properties characteristic of microscopic fungi of the genus Fusarium spp.
In our study, the multiplex PCR we detected among the 24 isolates isolated from corn grain obtained from Kyiv, Cherkasy, Zhytomyr and Chernihiv regions, 5 isolates were carrying the gene associated with the production of fumonisin (20.8%) and 2 (8.3%) isolates were associated with trichothecene production.
Conclusions and prospects for further research:
1. As a result of research, it was found that the use of multiplex PCR allows to determine the presence of toxin-producing microscopic fungi of the genus Fusarium.
2. Analysis of research results showed that among 24 isolates isolated from corn grain obtained from Kyiv, Cherkasy and Kherson regions, 5 isolates produced fumonisin (20.8%) and 2 (8.3%) isolates – trichothecene.
The use of PCR to detect toxin-producing microscopic fungi of the genus Fusarium in Ukraine is important for improving measures to prevent poisoning of animals with mycotoxins and improve the quality of livestock products.
Keywords: Fusarium, fumonisine, PCR, isolate, toxigenic strain.
- Vasjanovich, O.M., Korzunenko, O.F., & Obrazhej, A.F. (2003). Monіtoringovі doslіdzhennja mіkobіoti kormіv z rіznih regіonіv Ukrainy [Monitoring researches of mycobiota feeds from different regions of Ukraine]. Veterinarna bіotechnologіia – Veterinary Bіotechnology, 4, 27-30 [in Ukrainian].
- Boltjanskaja, Je.V., Kuvaeva, M.B., & Krojakova, E.A. (1989). Izuchenie rasprostranjonnosti toksigennyh shtammov gribov roda Fusarium i faktorov, vlijajushhih na ihtoksinoobrazovanie [Study of the prevalence of toxigenic strains of the fungus of the genus Fusarium and factors affecting their toxin formation]. Sbornik nauchnyh trudov – Collection of scientific papers, 9, 256-262 [in Russian].
- Burjak, V.N. (2007). Mikotoksikozy svinej i ih profilaktika [Mycotoxicoses of pigs and their prevention]. Zootehnija – Zootechnics, 9, 29-31 [in Russian].
- Apatenko, V. (2011). Nebezpechnі mіkotoksini [Dangerous mycotoxins]. Agrobіznes sogodni – Agribusiness today, 1-2, 18-20 [in Ukrainian].
- Satton, D., Fotergil, A., & Rinaldi, M. (2001). Opredelitel patogennykh i uslovno patogennykh gribov [Determinant of pathogenic and conditionally pathogenic fungy]. Moscow: Mir [in Russian].
- Malіnіn, O., Kutsan, O., & Shevcova, G. (2003). Mіkotoksikologіchnij monіtoring koncentrovanih kormіv Lіsostepu Ukraїni [Mycotoxicology monitoring of concentrated feeds of the forest steppe of Ukraine]. Tvarinnictvo Ukraini – Animal husbandry of Ukraine, 12, 26-28 [in Ukrainian].
- Dragan Y.P., Bidlack W.R., Cohen S.M., Goldsworthy T.L., Hard G.C., Howard P.C., et al. (2001). Implications of apoptosis for toxicity, carcinogenicity and risk assessment: fumonisin B1 as an example. Toxicol. Sci.,61, 6-17. https://doi.org/10.1093/toxsci/61.1.6.
- Grenier B., & Oswald I. (2011). Mycotoxin co-contamination of food and feed: meta-analysis of publications describing toxicological interactions. World Mycotoxin Journal, 4(3), 285-313. https://doi.org/10.3920/WMJ2011.1281.
- Henry, M.H., Wyatt, R.D., & Fletchert, O.J. (2000). The toxicity of purified fumonisin B1 to broiler chicks. Poultry Sci.,79, 1378-1384. https://doi.org/ 10.1093/ps/79.10.1378.
- Howard, P.C., Eppley, R.M., Stack, M.E., Warbritton, A., Voss, K.A., Lorentzen, R.J., Kovach, R.M., & Bucci, T.J. (2001). Fumonisin B1 carcinogenicity in a two-year feeding study using F344 rats and B6C3F1 mice. Environ. Health Persp., 109 (Suppl. 2), 277-282. https://doi.org/10.1289/ehp.01109s2277.
- Rahmani, A., Jinap, S., & Soleimany, F. (2009). Qualitative and Quantitative Analysis of Mycotoxins. Food Science And Food Safety., 8, 202-252. https://doi.org/10.1111/j.1541-4337.2009.00079.x.
- Ross, P.F., Rice, L.G., Osweiler, G.D., Nelson, P.E., Richard, J.L., & Wilson, T.M. (1992). A review and update of animal toxicoses associated with fumonisin-contaminated feeds and production of fumonisins by Fusarium isolates. Mycopathologia, 117, 109-114. https://doi.org/10.1007/BF00497286.
- Nelson, P.E., Desjardins A.E., & R.D. Plattner. (1993). Fumonisins, mycotoxins produced by Fusarium species; biology, chemistry, and signicance. Annu. Rev. Phytopathol., 31, 233-252. https://doi.org/10.1146/annurev.py.31.090193.001313.
- Bluhm, B.H., Flaherty, J.E., Cousin, M.A., & Woloshuk, C.P. (2002). Multiplex polymerase chain reaction assay for the differential detection of trichothecene- and fumonisin-producing species of Fusarium in cornmeal. J Food Prot.,65(12), 1955-61. doi: 10.4315/0362-028x-65.12.1955. https://doi.org/10.4315/0362-028x-65.12.1955.
Download full text in PDF