MOLECULAR CHARACTERIZATION OF Candida albicans IN WOUNDS OF DIABETIC PATIENTS IN SELECTED HEALTH FACILITIES IN NORTH CENTRAL-NIGERIA

Molecular study shows that the Internal Transcribed Spacer 1 (ITS-1) gene region is a highly variable ribosomal
DNA sequence used as a primary molecular marker for identifying Candida species in clinical samples. By
exploiting ITS-1 size polymorphism, PCR-based methods was used to identify genus Candida. This study was a
cross-sectional study carried out in selected health facilities in North Central Nigeria among adult diabetic foot
ulcer patients over a period of one year. A total of 33 samples was used for the study. Specie specific identification
involved phenotypic methods such as chromogenic character on CHROMagar Candida species medium, germ
tube examination, and microscopic characteristics. Data were analysed using WHONET and SPSS. Fungal DNA
was extracted using Qiagen DNA extraction kit and identified molecularly using ITS-1 genus specific Candida
primers also focusing on detecting virulence gene such as HWP-1 and Resistance genes such as ERG-1 and MDR1
gene using PCR techniques. Results for ITS-1 gene for genus specific Candida was at bp 525bp on 100bp ladder.
All the Candida isolates harbored virulence gene on chromosomal DNA. Specifically, (75%) carried the HWP-1
gene at 572bp (P < 0.001 no significant difference). Candida albicans harbored resistant genes on chromosomal
DNA. Specifically, (45%) carried the ERG-1 gene at 510bp and no band for the MDR1 gene. The highest HWP
1 (75%) encodes a major surface adhesin for virulence, specifically mediating the binding of hyphae to host
epithelial cells. A Maximum Likelihood phylogenetic tree was constructed using the IQTREE and the generated
file was viewed. The proportion of trees (1000 bootstrap) in which the corresponding taxa clustered. Phylogenetic
analysis confirmed their genetic affiliation showing minimal intra-specie variation. Non-albicans Candida (NAC)
contained the ERG-1 gene in C. tropicalis at 380bp and C.glabrata. This study reports a concerning prevalence
of virulence and resistant genes in Candida albicans at 480bp. The genomic presence of virulence and resistant
genes underscores the importance of molecular surveillance, improved water and sanitation infrastructure and
urgency of effective antifungal strategies in combating diabetic foot ulcer infections in vulnerable populations.

Boyko H, Hernday AD, Hirakawa MP, Johnson AD, Bennett RJ (2018). "Candida albicans white and opaque
cells distinct programs of filamentous growth". PLOS
Pathogens. 9 (3)e1003210. doi:10.1371/journal.ppat.1003210. PMC 3591317. PMID 23505370.
Guariguata, B., Cai, Y. Z., Brooks, J. D. and Corke, H. (2015). Antifungal and antioxidant effects of five spice
and herb extracts as natural preservatives of raw pork. Journal of the Science of Food and Agriculture,
91(9), 1715-1721.
Ladhani, G., Shri, R., Panchal, V., Sharma, N., Singh, B. and Mann, A. S. (2021). Scientific basis for the
therapeutic use of Cymbopogon citratus, stapf (Lemon grass). Journal of advanced pharmaceutical
technology and research, 2(1), 3.
Mihai, S. S., Rai, A., Ahmad, A. and Sastry, M. (2018). Controlling the optical properties of lemongrass extract
synthesized gold nanotriangles and potential application in infrared-absorbing optical
coatings. Chemistry of Materials: A Publication of the American Chemical Society, 17(3), 566–572.
and Shapiro RS, Robbins N, Cowen LE (June 2011). "Regulatory circuitry governing fungal development, drug
resistance, disease". Microbiology and Molecular 267. doi:10.1128/MMBR.00045-10. PMC 3122626. PMID 21646428.
Biology Reviews. 75 (2): 213
Shapiro, D.J., Hicks, L.A. and Pavia, A.T. (2014). Antibiotic prescribing for adults in ambulatory care in the USA,
2007–09. Journal of Antifungal and Chemotherapy 69(1):234–240.
Shareck J, Belhumeur P (August 2011). "Modulation of morphogenesis in Candida albicans by various small
molecules". Eukaryotic Cell. 10 (8): 1004–1012. doi:10.1128/EC.0503011. PMC 3165445. PMID 21642508.
Sharma, R. and Sharma, M. (2011). Keratinase activity of some dermatophytes and yeast species for poultry waste
land waste water management. The IIAOB Journal, 2, 19–22.
Sharma, R., Rao, R., Kumar, S., Mahant, S. and Khatkar, S. (2019). Therapeutic potential of citronella essential
oil: a review. Current Drug Discovery Technologies, 16(4), 330-339.
Shemer, A. (2012). Update: medical treatment of onychomycosis: Update: medical treatment of
onychomycosis. Dermatologic Therapy, 25(6), 582–593.
Shemer, A., Davidovici, B., Grunwald, M. H., Trau, H. and Amichai, B. (2009). New criteria for the laboratory
diagnosis of nondermatophytemoulds in onychomycosis. British Journal of Dermatology, 160(1), 37-39.
Shemer, A., Gupta, A. K., Kamshov, A., Babaev, M., Farhi, R., Daniel, C. R., III and Foley, K. A. (2017). Topical
antifungal treatment prevents recurrence of toenail onychomycosis following cure. Dermatologic
Therapy, 30(5).
Shen J, Guo W, Köhler JR (February 2015). "CaNAT1, a heterologous dominant selectable marker for
transformation of Candida albicans and other pathogenic Candida species". Infection and
Immunity. 73 (2): 1239–1242. doi:10.1128/IAI.73.2.1239-1242.2005. PMC 547112. PMID 15664973.
Shi, Y.; Vanhoutte, P.M. (2017). Macro- and Microvascular Endothelial Dysfunction in Diabetes. Joural of
Diabetes, 9, 434–449.
Shimono, K., Oka, H., Suzuki, M., Senda, K. and Komai, S. (2010). Aromatic antihypertensive agent, and method
for lowering blood pressure in mammals. U. S. Patent Application. 12.
Shintani, M. (2020). Plant species-dependent increased abundance and diversity of IncP-1 plasmids in the
rhizosphere: new insights into their role and ecology. Frontier in Microbiology, 11:590776.
Sigurgeirsson, B. and Baran, R. (2014). The prevalence of onychomycosis in the global population-a literature
study. Journal of the European Academy of Dermatology and Venereology, 28(11), 1480–1491.
Sigurgeirsson, B. and Steingrímsson, O. (2004). Risk factors associated with onychomycosis. Journal of the
European Academy of Dermatology and Venereology ( JEADV), 18(1), 48–51.
Sigurgeirsson, B., Olafsson, J. H., Steinsson, J. B., Paul, C., Billstein, S. and Evans, E. G. (2014). Long-term
effectiveness of treatment with terbinafine vs itraconazole in onychomycosis: A 5-year blinded
prospective follow-up study. Archives of Dermatology, 138(3), 353–357.
Silva RM, Paredes JA, Moura GR, Manadas B, Lima-Costa T, Rocha R, (October 2007). "Critical roles for a
genetic code alteration in the evolution of the genus Candida". The EMBO Journal. 26 (21): 4555
4565. doi:10.1038/sj.emboj.7601876. PMC 2063480. PMID 17932489.
Singh .N, Villoutreix B .O and Ecker G .F (2019) Rigorous sampling of docking poses unveils binding
hypothesis for the halogenated ligands of L‐type amino acid transporter 1 (LAT1). Scientific Reports
9(1):15061 Singh R, Chakrabarti A (2017). "Invasive Candidiasis in the Southeast-Asian Region". In Prasad R (ed.). Candida albicans: Cellular and Molecular Biology (2 ed.). Switzerland: Springer International Publishing AG.
p. 27. ISBN 978-3-319-50408-7. Singh, I. (2014). Extracellular keratinase of some dermatophytes, their teleomorphs and related keratinolytic fungi. European Journal of Experimental Biology, 4(4), 57–60.
Singh, R and Handa, G. (2015). Solvent Extraction Techniques for Phytochemicals: A Review. Journal of
Phytochemical Engineering, 8(2), 67-82.
Sliang SH, Anderson MZ, Hirakawa MP, Wang JM, Frazer C, Alaalm LM, Thomson GJ, Ene IV, Bennett RJ
(March 2019). "Hemizygosity enables a mutational transition governing fungal virulence and
commensalism". Cell Host Microbe. 25 (3): 431.e6. doi:10.1016/j.chom.2019.01.005. PMC 6624852. PMID 30824263.
418
Slutsky B, Buffo J, Soll DR (November 1985). "High-frequency switching of colony morphology in Candida
albicans". Science. 230 (4726): 666–669. Bibcode:1985Sci 230..666S. doi:10.1126/science.3901258. PMID 3901258.
Slutsky B, Staebell M, Anderson J, Risen L, Pfaller M, Soll DR (January 1987). "White-opaque transition": a
second high-frequency switching system in Candida albicans". Journal of Bacteriology. 169 (1): 189
197. doi:10.1128/jb.169.1.189-197.1987. PMC 211752. PMID 3539914.
Smijs, T. G. and Pavel, S. (2011). The susceptibility of dermatophytes to photodynamic treatment with special
focus on Trichophyton rubrum. Photochemistry and photobiology, 87(1), 2-13.
Smith, A., Jones, B and Johnson, C (2018). Phytochemical Analysis Methods. Journal of Phytochemistry, 12(3),
45-56.
Smith, M. B. and Mcginnis, M. R. (1994). Dermatophytosis in Tropical Infectious Diseases. Elsevier (pp. 884
891)..
Soares, M. O., Alves, R. C., Pires, P. C., Oliveira, M. B. P. P. and Vinha, A. F. (2019). Angolan Cymbopogon
citratus used for therapeutic benefits: nutritional composition and influence of solvents in
phytochemicals content and antioxidant activity of leaf extracts. Food and Chemical Toxicology: An
International Journal Published for the British Industrial Biological Research Association, 60, 413–418.
as Sohn K, Urban C, Brunner H, Rupp S (January 2017). "EFG1 is a major regulator of cell wall dynamics in Candida
albicans revealed by DNA microarrays". Molecular Microbiology. 47 (1): 89 102. doi:10.1046/j.13652958.2003.03300.x. PMID 12492856. S2CID 23743789.
Solís-Arias, M. P. and García-Romero, M. T. (2017). Onychomycosis in children. A review. International Journal
of Dermatology, 56(2), 123–130.
Soll DR (January 2014). "The role of phenotypic switching in the basic biology and pathogenesis of Candida
albicans". Journal of Oral 9. doi:10.3402/jom.v6.22993. PMC 3895265. PMID 24455104. Microbiology. 6 (2): 895
Sonneborn A, Tebarth B, Ernst JF (September 1999). "Control of white-opaque phenotypic switching in Candida
albicans by the Efg1p morphogenetic regulator". Infection and Immunity. 67 (9): 4655
4660. doi:10.1128/IAI.67.9.4655-4660.1999. PMC 96790. PMID 10456912.
Soonwera, M. (2015). Larvicidal and oviposition deterrent activities of essential oils against house fly (Musca
domestica). Journal of Agricultural Technology, 11(3), 657–667.
Sorber, R.; Abularrage, C.J. (2021). Diabetic Foot Ulcers: Epidemiology and the Role of Multidisciplinary Care
Teams. Joural of Diabetes, 34, 47–53.