Main Article Content



egg quality, Haugh units, micromycetes, PCR, storage, table eggs


The aim of this study was to monitor the quality and mycobiota composition of table eggs during storage period. The most significant changes in the egg weight and water activity were observed on Day 7. To identify the mycobiota present on the eggshell by PCR method, a newly designed procedure for the extraction of fungal DNA based on a combination of commercial isolation kit, proteinase K and ultrasound was implemented. Identified mold genera included Penicillium spp., Cladosporium spp., Fusarium spp. and Alternaria alternata group. Their ratio varied considerably during storage with the dominance of Penicillium spp. on Day 14.

Abstract 109 | pdf Downloads 102


Abd-Elsalam K.A., Aly I.N., Abdel-Satar M.A., Khalil, M.S. and Verreet J.A. 2003. PCR identification of Fusariumgenus based on nuclear ribosomal-DNA sequence data. African J. Biotechnol. 2(4):82.

Adams R.P., Miller J.S., Golenberg E.M. and Adams J.E.(Ed.). 1994.“Conservation of Plant Genes II: Utilization of Ancient and Modern DNA“ 2nd ed. Missouri Botanical Garden, Missouri, USA.

Akyurek H. and Okur A.A. 2009. Effect of storage time, temperature and hen age on egg quality in free-range layer hens. J. Anim. Vet. Advances. 8(10):1953.

Alexander C., Shankland G., Carman W. and Williams C. 2011. Introduction of a dermatophyte polymerase chain reaction assay to the diagnostic mycology service in Scotland. Br. J. Dermatol. 164(5):966.

Alleoni A.C.C. and Antunes A.J. 2004. Internal quality of eggs coated with whey protein concentrate. Scientia Agricola. 61(3):276.

Bahobail A.A.S., Hassan S.A. and El-Deeb B.A. 2012. Microbial quality and content aflatoxins of commercially available eggs in Taif, Saudi Arabia. Afr. J. Microbiol. Res. 6(13):3337.

Beifuss B., Bezold G., Gottlöber P., Borelli C., Wagener J., Schaller M. and Korting H.C. 2011. Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer. Mycoses. 54(2):137.

Bergmans A., van der Ent M., Klaassen N., Böhm G., Andriesse I. and Wintermans, R.G. 2010. Evaluation of a single-tube real-time PCR for detection and identification of 11 dermatophyte species in clinical material. Clin. Microbiol. Infect. 16(6):704.

Brillowska-Dabrowska A., Saunte D.M. and Arendrup M.C. 2007. Five-hour diagnosis of dermatophyte nail infections with specific detection of Trichophyton rubrum. J. Clin. Microbiol. 45(4):1200.

Brillowska-Dabrowska A., Nielsen S.S., Nielsen H.V. and Arendrup M.C. 2010a. Optimized 5-hour multiplex PCR test for the detection of tinea unguium: performance in a routine PCR laboratory. Med. Mycol. 48(6):828.

Brillowska-Dabrowska A., Swierkowska A., Lindhardt Saunte D.M. and Arendrup M.C. 2010b. Diagnostic PCR tests for Microsporum audouinii, M. Trichophyton infections. Med. Mycol. 48(3):486.

Caner C. 2005. The effect of edible eggshell coatings on egg quality and consumer perception. J. Sci. Food Agric. 85:1897.

Caner C. and Yüceer M. 2015. Efficacy of various protein-based coating on enhancing the shelf life of fresh eggs during storage. Poult. Sci. 94(7):1665.

Chelkowski J. and Visconti A. (Ed.).1992.“Alternaria: Biology, Plant Diseases and Metabolites“ 1sted. Elsevier, Amsterdam, The Netherlands.

Cupáková Š., Karpíšková R. and Necidová L. (Ed.). 2010. “Mikrobiologie potravin“ 1st ed. University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.

Cmoková A., Hamal P., Svobodová L. and Hubka V. 2014. Detection, identification and typing of dermatophytes by molecular genetic methods. Cesk. Dermatol. 89(4):175.

De Leo R., Quartieri A., Haghighi H., Gigliano S., Bedin E. and Pulvirenti A. 2018. Application of pectin-alginate and pectin-alginate-laurolyl arginate ethyl coatings to eliminate Salmonella enteritidis cross contamination in egg shells. J. Food Saf. 38(6):e12567.

De Reu K., Gryspeerdt K., Messens W., Heyndrickx M., Uyttendaele M., Debevere J. and Herman L. 2006. Eggshell factors influencing eggshell penetration and whole egg contamination by different bacteria, including Salmonella enteritidis. Int. J. Food Microbiol. 112(3):253.

Ebihara M., Makimura K., Sato K., Abe S. and Tsuboi R. 2009. Molecular detection of dermatophytes and nondermatophytes in onychomycosis by nested polymerase chain reaction based on 28S ribosomal RNA gene sequences. Br. J. Dermatol. 161(5):1038.

EC (European Communities). 2003. Commission Regulation (EC) No 2295/2003, 23 December, 2003.Council. Off. J. Eur. Union, L340, 16-34.

EC (European Communities). 2008. Commission Regulation (EC) No 589/2008, June 23, 2008. Council. Off. J. Eur. Union, L163, 6-23.

El Malt L.M. 2015. Assesment of the microbial quality and aflatoxins content in poultry farms eggs sold in Qena city-Upper Egypt. Assiut. Vet. Med. J. 145(61):144.

Erkme O. and Bozoglu T.F. (Ed). 2016.“Food Microbiology: Principles into Practice. Spoilage of Eggs and Egg Products“ 1st ed. John Wiley & Sons, Ltd., Manhattane, USA.

Faggi E., Pini G. and Campisi E. 2005. Use of magnetic beads to extract fungal DNA. Mycoses. 48(1):3.

Figueiredo T.C., Assis D.C.S., Menezes L.D.M., Oliveira D.D., Lima A.L., Souza M.R. and Cançado S.V. 2014. Effects of packaging, mineral oil coating, and storage time on biogenic amine levels and internal quality of eggs. Poult. Sci. 93(12):3171.

Fredricks D.N., Smith C. and Meier A. 2005. Comparison of six DNA extraction methods for recovery of fungal DNA as assessed by quantitative PCR. J. Clin. Microbiol. 43(10):5122.

Garg J., Tilak R., Garg A., Prakash P., Gulati A.K. and Nath G. 2009. Rapid detection of dermatophytes from skin and hair. BMC Res. Notes. 2(1):60.

Gilardi G., Demarchi S., Ortu G., Gullino M.L. and Garibaldi A. 2015. Occurrence of Alternaria japonica on seeds of wild and cultivated rocket. J. Phytophatol. 163(5):419.

Greco M.V., Franchi M.L., Golba S.L.R., Pardo A.G. and Pose G.N. 2014. Mycotoxins and mycotoxigenic fungi in poultry feed for food-producing animals. Scient World J. 2014:9.

Griffin D.W., Kellogg C.A., Peak K.K. and Shinn E.A. 2002. A rapid and efficient assay for extracting DNA from fungi. Lett. Appl. Microbiol. 34(3):210.

Hidalgo A., Rossi M. and Pompei C. 2006. Estimation of equivalent egg age through furosine analysis. Food Chem. 94:608.

Jin Y.H., Lee K.T. Lee W.I. and Han Y.K. 2011. Effects of storage temperature and time on the quality of eggs from laying hens at peak production. Asian-Australas. J. Anim. Sci. 24(2):279.

Jones D.R., Cox N.A., Guard J., Fedorka-Cray P.J., Buhr R.J., Gast R.K., Abdo Z., Rigsby L.L., Plumblee J.R., Karcher D.M., Robison C.I., Blatchford R.A. and Makagon M.M. 2015. Microbiological impact of three commercial laying hen housing systems. Poult. Sci. 94(3):544.

Kano R., Hirai A., Muramatsu M., Watari T. and Hasegawa A. 2003. Direct detection of dermatophytes in skin samples based on sequences of the chitin synthase 1 (CHS1) gene. J. Vet. Med. Sci. 65(2):267.

Karakousis A., Tan L., Ellis D., Alexiou H. and Wormald P.J. 2006. An assessment of the efficiency of fungal DNA extraction methods for maximizing the detection of medically important fungi using PCR. J. Microbiol. Methods. 65(1):38.

Karimiazar F., Soltanpour M.S., Aminzare M. and Hassanzadazar H. 2019. Prevalence, genotyping, serotyping, and antibiotic resistance of isolated Salmonella strains from industrial and local eggs in Iran. J. Food Saf. 39(1):e12585.

Kassambara A. and Mundt, F. 2007. Factoextra: Extract and Visualize the Results of Multivariate Data Analyses. R package version 1.0.5.

Kim J.Y., Choe Y.B., Ahn K.J. and Lee Y.W. 2011. Identification of dermatophytes using multiplex polymerase chain reaction. Ann. Dermatol. 23(3):304.

Kokkonen M., Ojala L., Parikka P. and Jestoi M. 2010. Mycotoxin production of selected Fusarium species at different culture conditions. Int.J. Food Microbiol. 143(1-2):17.

Kondori N., Abrahamsson A.L., Ataollahy N. and Wenneras C. 2010. Comparison of a new commercial test, Dermatophyte-PCR kit, with conventional methods for rapid detection and identification of Trichophyton rubrum in nail specimens. Med. Mycol. 48(7):1005.

Kurtzman C.P., Fell J.W. and Boekhout. T. (Ed.). 2011.“The Yeasts, a Taxonomic Study“ 5th ed. Elsevier, Amsterdam, Netherlands.

Lazar V. (Ed.). 1990.“Chov drubeže“ 1st ed. Mendel University, Brno, Czech Republic.

Lee S.B., Milgroom M.G. and Taylor J.W. 1988. A rapid, high yield mini-prep method for isolation of total genomic DNA from fungi. Fungal Genet. Rep. 35(1):23.

Lemfack M., Nickel J., Dunkel M., Preissner R. and Piechulla B. 2014. m VOC: a database of microbial volatiles. Nucleic Acids Res. 46(D1): D1261.

Li S. L., Zhou B., Yang L.Y. Li, Z.Y., Zhang Q. and Chen Y.W. 2002. An improved method for extracting fungal DNA. Journal of Yunnan University. 24(6):471.

Litz . and Cavagnolo R. 2010. Polymerase chain reaction in the diagnosis of onychomycosis: a large, single-institute study. Br. J. Dermatol. 163(3):511.

Lucisano M., Hidalgo A., Comelli E.M. and Rossi M. 1996. Evolution of chemical and physical albumen characteristics during the storage of shell eggs. J. Agric. Food Chem. 44:1235.

Martín I., García T., Fajardo V., Rojas M., Pegels N., Hernández P.E. and Martín I.G. 2009. SYBR-Green real-time PCR approach for the detection and quantification of pig DNA in feedstuffs. Meat Sci.82(2):252.

Matušovicová E., Žillová M. and Dekastellová L. (Ed.) 1986. “Technológia hydinárskeho priemyslu“ 1st ed. Príroda, Bratislava, Slovak Republic.

Mehlig L., Garve C., Ritschel A., Zeiler A., Brabetz W., Weber C. and Bauer A. 2014. Clinical evaluation of a novel commercial multiplex-based PCR diagnostic test for differential diagnosis of dermatomycoses. Mycoses 57(1):27.

Messens W., Grijspeerdt K., De Reu K., De Ketelaere B., Mertens K., Bamelis F., Kemps B., De Baerdemaeker J., Decuypere E. and Herman L. 2007. Eggshell penetration of various types of hens’ eggs by Salmonella entericaserovar Enteritidis. J. Food Prot. 70(3):623.

Míková K. and Davídek J. 2000. Criteria of quality and freshness of hen eggs. Czech J. Food Sci.18(6):250.

Morsy M.K., Sharoba A.M., Khalaf H.H., El-Tanahy H.H. and Cutter C.N. 2015. Efficacy of antimicrobial pullulan-based coating to improve internal quality and shelf-life of chicken eggs during storage. J. Food Sci. 80(5): M1066.

Mottet A. and Tempio G., 2017. Global poultry production: current state and future outlook and challenges. Worlds Poult. Sci. J. 73(2):245.

Nagy J., Jevinová P., Korenéková B., Kožárová I., Pipová M., Popelka P. and Turek P. (Ed.) 2011.“Technológia výroby a kvalita produktov hydiny, rýb, zveriny amedu“ 1st ed. Publishing Center of the University of Veterinary Medicine, Košice, Slovak Republic.

Neamatallah A.A., El-Leboudy A., Amer A.A. and El-Shenawy N.M. 2009. Biosafety against fungal contamination of hen's eggs and mycotoxins producing species. JKAU: Met., Env. & Arid Land Agric. Sci. 20(2):63.

Nedomová Š. 2012. Stanovení jakostních znaku skorápkových slepicích vajec. Thesis, Mendel University, Brno, Czech Republic.

Nedomová Š. and Simeonovová J. 2010. Effect of length and temperature of storage on the egg quality parameters. Slovak J. Food Sci. 4:196.

Obi C.N. and Igbokwe A.J. 2009. Microbiological analyses of freshly laid and stored domestic poultry eggs in selected poultry farms in Umuahia, Abia State, Nigeria. Res. J. Biol. Sci. 4(12):1297.

Pavón M.Á., González I., Pegels N., Martín R. and García T. 2010. PCR detection and identification of Alternariaspecies-groups in processed foods based on the genetic marker Alt a 1. Food Control. 21(12):1745.

Pedersen L.H., Skouboe P., Boysen M., Souleb J. and Rossen L.1997. Detection of Penicillium species in complex food samples using the polymerase chain reaction. Int. J. Food Microbiol. 35(2):169.

Pereira V.J., Fernandes D., Carvalho G., Benoliel M.J., San Romão M.V. and Barreto Crespo M.T. 2010. Assessment of the presence and dynamics of fungi in drinking water sources using cultural and molecular methods. Water Res. 44(17):4850.

R Core Team. 2018. R (Version 3.5.1.). Vienna, Austria: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing. Retrieved from www.r-project.org/

Rainieri S., Zambonelli C. and Kaneko Y. 2003. Saccharomyces sensu stricto: systematics, genetic diversity and evolution. J. Biosci. Bioeng. 96(1):1.

Rohweder D., Valenta H., Sondermann S., Schollenberger M., Drochner W., Pahlow G., Döll S and Dänicke S. 2011. Effect of different storage conditions on the mycotoxin contamination of Fusarium culmorum-infected and non-infected wheat straw. Mycotoxin Res. 27(2):145

Salihu M.D., Garba B. and Isah Y. 2015. Evaluation of microbial contents of table eggs at retail outlets in Sokoto metropolis, Nigeria. Sokoto J. Vet. Sci. 13(1):22.

Samli H.E., Agma A. and Senkoylu N. 2005. Effects of storage time and temperature on egg quality in old laying hens. J. Appl. Poult. Res.14(3):548.

Samson R.A. and Pitt J.I. (Ed.). 2000.“Integration of modern taxonomic methods for Penicillium and Aspergillus classification“ 1st ed. Harwood Academic Publishers, Reading, UK.

Sato T., Takayanagi A., Nagao K., Tomatsu N., Fukui T., Kawaguchi M., Kudoh J., Amagai M., Yamamoto N. and Shimizu N. 2010. Simple PCR-based DNA microarray system to identify human pathogenic fungi in skin. J. Clin. Microbiol. 48(7):2357.

Sebastien L., Josse J. and Husson F. 2008. Facto Mine R: An R package for multivariate analysis. J. Stat. Softw. 25(1):1.

Semjon B., Král M., Pospiech M., Reitznerová A., Malová J., Tremlová B. and Dudriková E. 2018. Application of multiple factor analysis for the descriptive sensory evaluation and instrumental measurements of bryndza cheese as affected by vacuum packaging. Int. J. Food Prop. 21(1):1508.

Simeonovová J. and Míková K. (Ed.). 2003.“Technologie drubeže,vajec a minoritních živo?išných produktu“ 1st ed. Mendel University, Brno, Czech Republic.

Simeonovová J., Míková K., Kubišová S. and Ingr I. (Ed.). 1999.“Technologie drubeže, vajec a minoritních živocišných produktu“ 1st ed. Mendel University, Brno, Czech Republic.

Simmons E.G. (Ed.). 2007.“Alternaria: An identification manual“ 6th ed. CBS Fungal Biodiversity Centre, Utrecht, The Netherlands.

Skóra J., Gutarowska B., Pielech-Przybylska K., Stepien L., Pietrzak K., Piotrowska M. and Pietrowski P. 2015 Assessment of microbiological contamination in the work environments of museums, archives and libraries. Aerobiologia. 31(3):389.

Steinhauserová I., Simeonovová J., Nápravníková E. and Tremlová B. (Ed.). 2003.“Produkce a zpracování drubeže, vajec amedu“ 1st ed. University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.

STN ISO 21527-1. 2010. Microbiology of food and animal feeding stuffs.Horizontal method for the enumeration of yeasts and molds.Part 1:Colony count technique in products with water activity greater than 0,95 (ISO 21527-1:2008). Slovak Standards Institute, Bratislava, Slovak Republic.

STN ISO 21527-2. 2010. Microbiology of food and animal feeding stuffs.Horizontal method for the enumeration of yeasts and molds. Part 2: Colony count technique in products with water activity less than or equal to 0,95 (ISO 21527-2:2008). Slovak Standards Institute, Bratislava, Slovak Republic.

STN EN ISO 6887-1. 2017. Microbiology of the food chain -Preparation of test samples, initial suspension and decimal dilutions for microbiological examination. Part 1: General rules for the preparation of the initial suspension and decimal dilutions (ISO 6887-1:2017). Slovak Standards Institute, Bratislava, Slovak Republic.

Sypecka Z., Kelly M. and Brereton P. 2004. Deoxynivalenol and zearalenone residues in eggs of laying hens fed with a naturally contaminated diet, effects on egg production and estimation of transmission rates from feed to eggs. J. Agric. Food Chem. 52(17):5463.

Thrane U., Adler A., Clase P.E., Galvano, F., Langseth W., Lew H., Logrieco A., Nielsen K.F. and Ritieni A. 2004. Diversity in metabolite production by Fusarium langsethiae, Fusarium poae and Fusarium sporotrichioides. Int. J. Food Microbiol. 95(3):257.

Tolik D., Polawska E., Charuta A., Nowaczewski S. and Cooper R. 2014. Characteristics of egg parts, chemical composition and nutritive value of Japanese quail eggs-a review. Folia Biol. (Krakow). 62(4):287.

Tomczyk l., Stepien l., Urbaniak M., Szablewski T., Cegielska-Radziejewska R. and Stuper-Szablewska K. 2018. Characterisation of the mycobiota on the shell surface of table eggs acquired from different egg-laying hen breeding systems. Toxins. 10(7):293.

Tomczyk L., Szablewski T., Stuper-Szablewska K., Nowaczewski S. and Cegielska-Radziejewska R. 2019. The influence of the conditions of acquisition and storage of table eggs on changes in their quality and the presence of mycobiota and Fusarium mycotoxins. Poult. Sci. 98(7):2964.

Uchida T., Makimura K., Ishihara K., Goto H., Tajiri Y., Okuma M., Fujisaki R., Uchida K., Abe S. and Iijima M. 2009. Comparative study of direct polymerase chain reaction, microscopic examination and culture-based morphological methods for detection and identification of dermatophytes in nail and skin samples. J. Dermatol. 36(4):202.

U.S. Department of Agriculture (USDA). U.S. Standards, Grades, and Weight Classes for Shell Eggs, 56.200 et seq. USDA Agricultural Marketing Service: Washington, DC, 1995.

Weidenborner M., Wieczorek C. and Kunz B. 1995. Mold spectra of various foods in relation to plating medium. J. Food Prot. 58(6):661.

Wisselink G., van Zanten E. and Kooistra-Smid A. 2011. Trapped in keratin; a comparison of dermatophyte detection in nail, skin and hair samples directly from clinical samples using culture and real-time PCR. J. Microbiol. Methods. 85(1):62.

Wong S.F., Mak J.W. and Pook P.C. 2007. New mechanical disruption method for extraction of whole cell protein from Candida albicans. Southeast Asian J. Trop. Med. Public Health 38(3):512.

Wu Z.H., Wang T.H., Huang W. and Qu Y.B. 2001. A simplified method for chromosome DNA preparation from filamentous fungi. Mycosystema. 20(4):575.

Xue S.J., Yue T.L., Guan J., Yuan Y.H. and Gao Z.P. 2006. An improved method for extracting fungal DNA. Food Res. Develop. 4:39.

Yano S., Koyabashi K. and Kato K. 2003. Intrabronchial lesion due to Cladosporium sphaerospermumin a healthy, non-asthmatic woman. Mycoses. 46(8):348.

Yüceer M. and Caner C. 2014. Antimicrobial lysozyme?chitosan coatings affect functional properties and shelf life of chicken eggs during storage. J. Sci. Food Agric. 94(1):153.

Zambrowicz A., Eckert E., Pokora M., Dabrowska A., Szoltysik M., Bobak L., Trziszka T. and Chrzanowska J. 2015. Biological activity of egg-yolk protein by-product hydrolysates obtained with the use of non-commercial plant protease. Ital. J. Food Sci. 27(4):450.

Zeng Q.Y., Westermark S.O., Lestander R. and Wang X.R. 2006. Detection and quantificaion of Cladosporium in aerosols by real-time PCR. J. Environ. Monit. 8(1):153.