Yaprağı Yenen Sebzelerdeki Mikroorganizma Türlerinin MALDI-TOF MS (Matris Destekli Lazer Desorpsiyon/İyonizasyon Uçuş Süresi Kütle Spektrometresi) Tekniği Kullanılarak Tanılanması

Aysun Uysul; Şener Kurt; Soner Soylu; Emine Mine Soylu; Merve Kara

doi:10.29133/yyutbd.627850

Araştırma Makalesi

Yaprağı Yenen Sebzelerdeki Mikroorganizma Türlerinin MALDI-TOF MS (Matris Destekli Lazer Desorpsiyon/İyonizasyon Uçuş Süresi Kütle Spektrometresi) Tekniği Kullanılarak Tanılanması

Yıl 2019, Cilt: 29 Sayı: 4, 595 - 603, 31.12.2019

Aysun Uysul Şener Kurt Soner Soylu Emine Mine Soylu Merve Kara

https://doi.org/10.29133/yyutbd.627850

Cited By: 9

Öz

Gıda
kaynaklı fungal ve bakteriyel patojenler giderek taze meyve ve sebzelerle
ilişkilendirilmektedir. Taze üretilen bitkilerin bakteriyel kolonizasyonu, dış
dokuda yüksek seviyelerde oluşabilir, ancak bitki dokusu içerisinde bakteriler
de tespit edilmektedir. Hatay ilinin Defne ve Antakya ilçelerinde farklı sebze
pazarlarından marul, ıspanak, lahana, roka, nane, maydanoz, dereotu, pırasa
gibi yapraklı sebzeler toplanmıştır. Mikrobiyolojik analizler için patates
dekstroz agar, nutrient agar ve kanlı agar besi yerleri kullanılmıştır. Bu
mikroorganizmaların tanılanması, fenotipik karakterler ve matris destekli lazer
desorpsiyon/iyonizasyon uçuş süresi kütle spektrometresi (MALDI-TOF MS)
tekniğine dayalı olarak gerçekleştirilmiştir. MALDI-TOF MS analizi sonucuna
göre mikroorganizmaların dağılımı incelendiğinde, tür düzeyinde 60 adet ile en
fazla bakteri türleri tanılanırken, bunu 4 adet fungus ve 1 adet maya
izlemiştir. En fazla mikroorganizma türü marul örneklerinden elde edilmiştir. Cins
düzeyinde dağılımları incelendiğinde; 9 tür ile Pseudomonas başta gelirken, bunu 5 tür ile Lactobacillus, 4 tür ile Acinetobacter
ve 2 tür ile Klebsiella, Citrobacter,
Enterobacter, Lysinibacillus, Pantoea 1 tür Bacillus, 1 tür Escherichia
coli, 1 tür Exiguobacterium
cinsleri izlemiştir. Mikroorganizma türlerinin koloni yoğunlukları
incelendiğinde, en yoğun koloni Pseudomonas
ve Acinetobacter cinsine ait türlerde
belirlenmiştir. En az koloni yoğunluğu funguslarda gözlenmiştir. Çalışma
sonucunda Salmonella sp, Klebsiella pneumoniae, Escherichia coli, Bacillus cereus, Klebsiella oxytoca türleri insan sağlığı için en
tehlikeli türler olarak bulunmuştur.

Anahtar Kelimeler

MALDI-TOF MS, Mikroorganizma, Taze sebze, Tanı

Destekleyen Kurum

Hatay Mustafa Kemal Üniversitesi Bilimsel Araştırma Projeleri Komisyon Başkanlığı

Proje Numarası

18.M.026

Teşekkür

Bu çalışma, Hatay Mustafa Kemal Üniversitesi Bilimsel Araştırma Projeleri Komisyon Başkanlığına tarafından 18.M.026 nolu proje kapsamında desteklenmiş ve Hatay Mustafa Kemal Üniversitesi Bitki Sağlığı Kliniği Uygulama ve Araştırma Merkezi laboratuvarlarında yürütülmüştür.

Kaynakça

Alam, MS. Feroz, F. Rahman, H. Das, K.K. Noor, R. (2015). Microbiological contamination sources of freshly cultivated vegetables, Nutrition & Food Science Vol. 45 No. 4, pp. 646-658.
Ayhan, T., Ufuk, B., and Sine, O.T. (2011). Efficacy of Different Decontamination Treatment on Microbial Population of Leafy Vegetables. Journal of Geographic Information and Decision Analysis. 36(1): 9-13.
Barak, J.D., Schroeder, B.K., 2012. Interrelationships of Food Safety and Plant Pathology: The Life Cycle of Human Pathogens on Plants. Annual Review of Phytopathology 50: 241-266.
Beuchat, LR. (2002). Ecological factors influencing survival and growth of human pathogens on raw fruits and vegetables. Microbes and Infection. 4: 413-423.
Carolis, E. D., Posteraro, B., Lass-Flo, C., Vella, A., Florio, A. R., Torelli, R., Girmenia, C., Colozza, C., Tortorano, A. M., Sanguinetti M., Fadda, G., (2012). Species identification of Aspergillus, Fusarium and Mucorales with direct surface analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clinic Microbiology Infect. 18: 475–484.
Chalupová, J., Raus, M., Sedlářová, M., Šebela, M., (2014). Identification of fungal microorganisms by MALDI-TOF mass spectrometry. Biotechnology Advances 32:230–241.
Gauthier, G.M., Keller, N.P., (2013). Crossover fungal pathogens: The biology and pathogenesis of fungi capable of crossing kingdoms to infect plants and humans. Fungal Genetics and Biology 61: 146–157.
Hasnain S, Rehman Y, Mahmood S (2015) Cross-Kingdom Pathogenicity across Plants and Human Beings. J Bacteriol Parasitol 6:e124. doi:10.4172/2155-9597.1000e124
Hausdorf L., Mundt K., Winzer, M., Cordes, C., Fröhling, A., Schlüter, O., Klocke, M., (2013). Characterization of the cultivable microbial community in a spinach-processing plant using MALDI-TOF MS. Food Microbiology. Food Microbiology 34 (2013) 406-411.
Kara, M., Uysal, A.,Soylu, S., Kurt, Ş., Soylu, E.M., (2017). Identification of Plant-Associated Microorganisms Employing MALDI TOF Mass Spectrometry as a Rapid Detection Technique. International Conference on Agriculture, Forest, Food Sciences and Technologies (ICAFOF), 15-17 May, 2017. Nevşehir, p. 1111.
Kirzinger, MW., Nadarasah, G., Stavrinides, J. (2011). Insights into crosskingdom plant pathogenic bacteria. Genes 2: 980-997.
Lund, B.M., Baird-Parker, T.C., and Gould, G.W. 2000. “The Microbiological Safety And Quality of Food.”Volume I. An Aspen Publictions, Maryland,
Mahmood, S., Rehman, Y., Hasnain, S., (2015). Cross-Kingdom Pathogenicity across Plants and Human Beings. J Bacteriol Parasitology 6: 4. doi:10.4172/2155-9597.1000-124.
Mellmann, A., Harmsen, D., Cummings, CA., Zentz, EB., Leopold, SR., (2011). Prospective Genomic Characterization of the German Enterohemorrhagic Escherichia coli O104:H4 Outbreak by Rapid Next Generation Sequencing Technology. Microbiology 105 (6), 1729-1743.
Roberts, T.A., Pitt, J.I., Farkas, J., and Graw, F.H. (1998). “Microorganisms in food. Microbial ecology of food commodities.” Blackie Academic & Professional, UK.
Sengun, I.Y., Karapinar, M.,( 2004). Effectiveness of lemon juice, vinegar and their mixture in the elimination of Salmonella typhimurium on carrots (Daucus carota L.). International Journal of Food Microbiology., 96: 301-305.
Sharma, K., Goss, E.M., Dickstein, E.R., Smith, M.E., Johnson, J.A., (2014). Exserohilum rostratum: characterization of a cross-kingdom pathogen of plants and humans. Plos One 9: 108691.
Sivapalasingam, S., Friedman, C. R., Cohen, L., Tauxe, R. V., 2004. Fresh produce: a growing cause of outbreaks of foodborne illness in the United States, 1973 through 1997. J. Food Prot. 67(10):2342-2353.
Uysal, A., Kurt, Ş., Soylu E.M., Kara M., Soylu S., (2018). Evaluation of the Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight (MALDI-TOF) Mass Spectrometry for Identification of Some Plant Fungal Pathogenic Species. International Agricultural Science Congress. 09-12 May Van/Turkey.
Vandekinderen, I., Van Camp, J., Devlieghere, F., Veramme, K., Denon, Q., Ragaert, P., De Meulenaer, B. (2008). Effect of decontamination agents on the microbial population, sensorial quality, and nutrient content of grated carrots (Daucus carota L.). Journal of Agricultural Food Chemistry.56: 5723-5731.
Wright, K. M., Chapman, S., McGeachy, K., Humphris, S., Campbell, E., Toth, I. K., and Holden, N. J. (2013). The endophytic lifestyle of Escherichia coli O157:H7: Quantification and internal localization in roots. Phytopathology 103:333-340.

Identification of Microorganism Species in Leafy Vegetables Using MALDI-TOF MS (Matrix Assisted Laser Desorption / Ionization Mass Spectrometry) Technique

Yıl 2019, Cilt: 29 Sayı: 4, 595 - 603, 31.12.2019

Aysun Uysul Şener Kurt Soner Soylu Emine Mine Soylu Merve Kara

https://doi.org/10.29133/yyutbd.627850

Cited By: 9

Öz

The
foodborne fungal and bacterial pathogens are increasingly associated with fresh
fruit and vegetables. Bacterial colonization of fresh produce plants can occur
to high levels on the external tissue but bacteria have also been detected
within plant tissue. Leafy vegetables such as lettuce, spinach, cabbage,
rocket, mint, parsley, dill, and leek were collected from different vegetable
markets in the counties of Defne and Antakya of Hatay province. Growing media potato
dextrose agar, nutrient agar and blood agar media were used for microbiological
analysis. Their identification were performed based on phenotypic characters
and matrix-assisted laser desorption/ionization time-of-flight mass
spectrometry (MALDI-TOF MS) technique. According to the results of MALDI-TOF MS
analysis, when the distribution of microorganisms were examined, the most
bacterial species were identified with 60 species at the species level,
followed by 4 fungi and 1 yeast. The
most microorganism species were obtained from lettuce samples. When the distribution of species level is
examined; firstly, 9 species with Pseudomonas,
5 species with Lactobacillus, 4
species with Acinetobacter and 2
species with Klebsiella, Citrobacter,
Enterobacter, Lysinibacillus were obtained and Pantoea one species Bacillus,
one species Escherichia coli, one
species followed by Exiguobacterium genera.
When the colony density of microorganism species were examined, the most
intensive colony was determined in the species belonging to the genus Pseudomonas and Acinetobacter. At least colony density was observed in fungi. In
result, Salmonella sp, Klebsiella pneumoniae, Escherichia coli,
Bacillus cereus, Klebsiella oxytoca species were found to be very dangerous
species for human health.

Anahtar Kelimeler

MALDI-TOF MS, Microorganism, fresh vegetables, identification

Proje Numarası

18.M.026

Kaynakça

Alam, MS. Feroz, F. Rahman, H. Das, K.K. Noor, R. (2015). Microbiological contamination sources of freshly cultivated vegetables, Nutrition & Food Science Vol. 45 No. 4, pp. 646-658.
Ayhan, T., Ufuk, B., and Sine, O.T. (2011). Efficacy of Different Decontamination Treatment on Microbial Population of Leafy Vegetables. Journal of Geographic Information and Decision Analysis. 36(1): 9-13.
Barak, J.D., Schroeder, B.K., 2012. Interrelationships of Food Safety and Plant Pathology: The Life Cycle of Human Pathogens on Plants. Annual Review of Phytopathology 50: 241-266.
Beuchat, LR. (2002). Ecological factors influencing survival and growth of human pathogens on raw fruits and vegetables. Microbes and Infection. 4: 413-423.
Carolis, E. D., Posteraro, B., Lass-Flo, C., Vella, A., Florio, A. R., Torelli, R., Girmenia, C., Colozza, C., Tortorano, A. M., Sanguinetti M., Fadda, G., (2012). Species identification of Aspergillus, Fusarium and Mucorales with direct surface analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clinic Microbiology Infect. 18: 475–484.
Chalupová, J., Raus, M., Sedlářová, M., Šebela, M., (2014). Identification of fungal microorganisms by MALDI-TOF mass spectrometry. Biotechnology Advances 32:230–241.
Gauthier, G.M., Keller, N.P., (2013). Crossover fungal pathogens: The biology and pathogenesis of fungi capable of crossing kingdoms to infect plants and humans. Fungal Genetics and Biology 61: 146–157.
Hasnain S, Rehman Y, Mahmood S (2015) Cross-Kingdom Pathogenicity across Plants and Human Beings. J Bacteriol Parasitol 6:e124. doi:10.4172/2155-9597.1000e124
Hausdorf L., Mundt K., Winzer, M., Cordes, C., Fröhling, A., Schlüter, O., Klocke, M., (2013). Characterization of the cultivable microbial community in a spinach-processing plant using MALDI-TOF MS. Food Microbiology. Food Microbiology 34 (2013) 406-411.
Kara, M., Uysal, A.,Soylu, S., Kurt, Ş., Soylu, E.M., (2017). Identification of Plant-Associated Microorganisms Employing MALDI TOF Mass Spectrometry as a Rapid Detection Technique. International Conference on Agriculture, Forest, Food Sciences and Technologies (ICAFOF), 15-17 May, 2017. Nevşehir, p. 1111.
Kirzinger, MW., Nadarasah, G., Stavrinides, J. (2011). Insights into crosskingdom plant pathogenic bacteria. Genes 2: 980-997.
Lund, B.M., Baird-Parker, T.C., and Gould, G.W. 2000. “The Microbiological Safety And Quality of Food.”Volume I. An Aspen Publictions, Maryland,
Mahmood, S., Rehman, Y., Hasnain, S., (2015). Cross-Kingdom Pathogenicity across Plants and Human Beings. J Bacteriol Parasitology 6: 4. doi:10.4172/2155-9597.1000-124.
Mellmann, A., Harmsen, D., Cummings, CA., Zentz, EB., Leopold, SR., (2011). Prospective Genomic Characterization of the German Enterohemorrhagic Escherichia coli O104:H4 Outbreak by Rapid Next Generation Sequencing Technology. Microbiology 105 (6), 1729-1743.
Roberts, T.A., Pitt, J.I., Farkas, J., and Graw, F.H. (1998). “Microorganisms in food. Microbial ecology of food commodities.” Blackie Academic & Professional, UK.
Sengun, I.Y., Karapinar, M.,( 2004). Effectiveness of lemon juice, vinegar and their mixture in the elimination of Salmonella typhimurium on carrots (Daucus carota L.). International Journal of Food Microbiology., 96: 301-305.
Sharma, K., Goss, E.M., Dickstein, E.R., Smith, M.E., Johnson, J.A., (2014). Exserohilum rostratum: characterization of a cross-kingdom pathogen of plants and humans. Plos One 9: 108691.
Sivapalasingam, S., Friedman, C. R., Cohen, L., Tauxe, R. V., 2004. Fresh produce: a growing cause of outbreaks of foodborne illness in the United States, 1973 through 1997. J. Food Prot. 67(10):2342-2353.
Uysal, A., Kurt, Ş., Soylu E.M., Kara M., Soylu S., (2018). Evaluation of the Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight (MALDI-TOF) Mass Spectrometry for Identification of Some Plant Fungal Pathogenic Species. International Agricultural Science Congress. 09-12 May Van/Turkey.
Vandekinderen, I., Van Camp, J., Devlieghere, F., Veramme, K., Denon, Q., Ragaert, P., De Meulenaer, B. (2008). Effect of decontamination agents on the microbial population, sensorial quality, and nutrient content of grated carrots (Daucus carota L.). Journal of Agricultural Food Chemistry.56: 5723-5731.
Wright, K. M., Chapman, S., McGeachy, K., Humphris, S., Campbell, E., Toth, I. K., and Holden, N. J. (2013). The endophytic lifestyle of Escherichia coli O157:H7: Quantification and internal localization in roots. Phytopathology 103:333-340.

Toplam 21 adet kaynakça vardır.

Ayrıntılar

Birincil Dil	Türkçe
Konular	Ziraat, Veterinerlik ve Gıda Bilimleri
Bölüm	Makaleler
Yazarlar	Aysun Uysul Bu kişi benim Şener Kurt Bu kişi benim 0000-0003-4545-5968 Soner Soylu 0000-0003-1002-8958 Emine Mine Soylu Bu kişi benim 0000-0001-5961-0848 Merve Kara Bu kişi benim 0000-0001-7320-3376
Proje Numarası	18.M.026
Yayımlanma Tarihi	31 Aralık 2019
Kabul Tarihi	15 Kasım 2019
Yayımlandığı Sayı	Yıl 2019 Cilt: 29 Sayı: 4

Kaynak Göster

APA	Uysul, A., Kurt, Ş., Soylu, S., Soylu, E. M., vd. (2019). Yaprağı Yenen Sebzelerdeki Mikroorganizma Türlerinin MALDI-TOF MS (Matris Destekli Lazer Desorpsiyon/İyonizasyon Uçuş Süresi Kütle Spektrometresi) Tekniği Kullanılarak Tanılanması. Yuzuncu Yıl University Journal of Agricultural Sciences, 29(4), 595-603. https://doi.org/10.29133/yyutbd.627850