ABSTRACT: 

This research aimed to explore the identification of the Phenotype and genotype of Debaryomyces species isolated from patients with diabetic foot in Al-Najaf City. Samples were collected from the depths of wounds in diabetic foot patients, ensuring aseptic precautions were taken, and then cultured on Sabouraud’s dextrose agar, followed by incubation at 30-35 °C for a duration of 5-7 days to promote growth. The identification of colonies was based on their microscopic characteristics. Subsequently, these yeasts were grown in chrome agar to isolate and identify Debaryomyces spp. A PCR technique was conducted to magnify the ITS1 and ITS4 sequences for the diagnosis of Debaryomyces spp., and the PCR products were exposed to sequencing using the Sanger method. All samples isolated were grown on SDA, and when utilizing CHROMagar differential agar. The findings indicated various molecular sizes of the ITS region for Debaryomyces spp., with the primer pair (ITS1-ITS4) targeting these ITS regions for the same sample. The products of PCR were dispatched to Macrogen Lab in the USA, where one sample of replicated products of PCR (both reverse and forward strands) was analyzed. The sequences in the current study were then matched with reference global sequences. It can be concluded that Debaryomyces species emerge as the most prevalent fungal pathogens associated with life-threatening invasive infections in highly immunocompromised individuals or those who have had major trauma and prolonged stay in the intensive care units. They occupy the fourth place as the most common agents of nosocomial-acquired bloodstream infections in hospitals.

REFERENCES :

1) Carmona, E.M. and Limper, A.H. (2017). Overview of treatment approaches for fungal infections. Clin Chest Med. 38(3):393-402.2.
2) Prista, C.; Michan, C.; Miranda, I.M.; Ramos, J. The halo tolerant Debaryomyceshansenii, the Cinderella of non-conventional yeasts. Yeast 2016, 33, 523–533.
3) Breuer, U.; Harms, H. Debaryomyceshansenii-extremophilic yeast with biotechnological potential. Yeast 2006, 23, 415–437.
4) Sánchez, N.S.; Calahorra, M.; González-Hernández, J.C.; Peña, A. Glycolytic sequence and respiration of Debaryomyceshansenii compared to Saccharomyces cerevisiae. Yeast 2006, 23, 361–374.
5) Navarrete, C.; Frost, A.T.; Ramos-Moreno, L.; Krum, M.R.; Martínez, J.L. A physiological characterization in controlled bioreactors reveals a novel survival strategy for Debaryomyceshansenii at high salinity. Yeast 2021, 38, 302–315.
6) Carvalheiro, F.; Duarte, L.C.; Lopes, S.; Parajó, J.C.; Pereira, H.; Gírio, F.M. Supplementation requirements of brewery’s spent grain hydrolysate for biomass and xylitol production by Debaryomyceshansenii CCMI 941. J. Ind. Microbiol. Biotechnol. 2006, 33, 646–654.
7) Axelson-Fisk, M.; Sunnerhagen, P. Comparative genomics and gene finding in fungi. In Methods in Molecular Biology; Springer: Berlin/Heidelberg, Germany, 2006; Volume 15, pp. 1–28.
8) Corredor, M.; Davila, A.-M.; Casarégola, S.; Gaillardin, C. Chromosomal polymorphism in the yeast species Debaryomyceshansenii. Antonie van Leeuwenhoek 2003, 84, 81–88.
9) Ramos, J.; Melero, Y.; Ramos-Moreno, L.; Michán, C.; Cabezas, L. Debaryomyceshansenii Strains from Valle De Los Pedroches Iberian Dry Meat Products: Isolation, Identification, Characterization, and Selection for Starter Cultures. J. Microbiol. Biotechnol. 2017, 27, 1576–1585.
10) Pisa D, Ramos M, Garcia P, et al. Fungal infection in patients with serpiginous choroiditis or acute zonal occult outer retinopathy, J ClinMicrobiol2008, vol. 46(pg. 130-5).
11) White, T.J.; Burns, T.D.; Lee, S.B. and Taylor, J.W. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, In PCR Protocols: A Guide to Methods and Applications, pp. 315–322. Edited by M. A. Innis, D. H. Gelfand, J. J. Sninsky, and T. J. White. San Diego, CA: Academic Press.
12) Katarzyna, G. (2011). Characteristics of growth of yeasts and yeast-like fungi on Chromogenic medium CHROMagar® Candida (GRASO). Department of Mycology, University of Warmia and Mazury, 1A Oczapowskiego Street, 10-957 Olsztyn, Poland; Wiadomooeci Parazytologiczne 2011, 57(3), 143–149.
13) Iyampillia T, Micheal JS, Mathai, E, and Mathews, MS. (2004) .Use of CHROMagar medium in the differentiation of Candida species: is it cost effective in developing countries. Trop.Med.Parasitol.98:279282.
14) Bellemain, E, Tor, C, Christian, B, Eric, C, Pierre, T, Havard, H. (2010). IT Searcas artainle environmental DNA barcode for fungi: aninsilico approach reveals potential PCR biases, BMC Microbiology, 10: 189,
http://www.biomedcentral.com/1471-2180/10/189.