Main Article Content
The current study included isolation of yeasts from the leaves of different types of citrus trees, and it was diagnosed based on phenotypic, culture and biochemical tests. The results showed that the yeasts belong to the species: Saccharomyces cerevisiae and Debaromyces hansenii. Also, Escherichia coli bacteria were obtained from the exit of infants in Ibn Al-Atheer Teaching Hospital in Mosul/ Iraq and was diagnosed based on biochemical tests. The resistance of isolates was studied for 20 antibiotics and 8 heavy metal ions. The results showed that S. cerevisiae is resistant to antibiotics (Amoxicillin, Ampicillin, Cephalexin monohydrate, Chloramphenicol, Clindamycin, Clotrimazol, Erythromycin, Getamicin, Newmycin, Penicillin, Tetracycline, and Vancomycin) and sensitive to Candizol, Fluconazole, Ketoconazol, Lamisil, Nystatin, Rifampin, Streptomycin, and Trimethoprim. While D. hansenii showed resistance to all antibiotics except Ampicillin, Candizol, Clindamycin, Clotrimazol, Erythromycin, Newmycin, Penicillin, and Tetracycline. The resistance of the two insulators differed towards heavy metal ions, as the two insulators were resistant to Nickel Chloride and Cobalt Chloride ions and Copper Sulfate sensitive. The ability of bacteria to resist antibiotics and heavy metal ions has been studied. The results showed that it showed sensitivity to Amoxicillin, Cephalexin monohydrate, Chloramphenicol, Getamicin, Newmycin, Nystatin and Streptomycin and resistance to the rest of the antibiotics used, while it was resistant to all heavy metals except for Cadmium Chloride, Mercury Chloride, and Zinc Sulfate.
Curing of plasmid DNA content of S. cerevisiae was succeed. The curred colonies showed sensitivity to the studied antibiotics at 15-84%, except for Ampicillin, Erythromycin, Lamisil and Vancomycin. The results of curing E. coli also showed a loss of antibiotic resistance in the range of 16-50%.
The conjugation between D. hansenii and S. cervisiae was successful as a acceptor with a conjugation frequency of 0.68×10-8. This study demonstrated that the transported plasmid DNA from D. hansenii carries antibiotic resistance Clotrimazol genes. In addition, an attempt was made to cross-kingdom conjugation between the yeast D. hansenii as a donor and E. coli as receptor. The results showed that the plasmid DNA carrying the Nystatin resistance genes has the ability to move and move from yeast to bacteria with a conjugation frequency of 2.05 × 10-8. The results also showed that the cured bacteria have the ability to receive and localize the plasmid from the yeast through the conjugation process.
Obaeda AEA, Mahmood AA. Fundamentals of biotechnology, modern knowledge library, Alexandria, Egypt. 2012;215.
Phillip EP. Biology, 2nd edition. Library of Congress. 2001;136.
Talaro K, Talaro A. Foundations in microbiology. Wm. C. Brown Publishers, Dubuque, U.S.A.; 1996.
Al-Faisal Abdul Hussein. Genetic engineering (1st ed.). Sunrise House for Publishing and Distribution, Amman, Jordan; 1999.
Hanson PK, Malone L, Birchmore JL, Nichols JW. Lem 3p is essential for the uptake and potency of alkylphosphocholine drugs, edelfosine and milte fosine. J. Biol. Chem. 2003;278:36041-50.
Mickelsen PA, Plorde JJ, Gordon KP, Hargiss C, Muclure J, Schoenknecht FD, Condie F, Tenover FC, Tompkins LS. Instability of antibiotics resistance in a strain of Staphylococcus epidermidis isolated from an outbreak of prosthetic valve endocarditis. J. Infect. Dis., Microbiology. Mosby publishing company, U.S.A. 1985;209(152):50-58.
Cramer R, Julian ED, Ralf H. Plasmid curing and generation of mutations induced with ethidium bromide in Stryptomycetes. Printed in Britain, J. of General Microbiology. 1986;132:819-824.
Pozieitner E, Zechner EL, Renner W, Fratte R, Jauk B, Hogenauer G, Koraiman G. Tra M of plasmid R1 controls trans fergene expression as an integrated control elementin a complex regulatory network. Mol. Microbiol. 1997;25(3):495-507.
Fritz HK, Kurt AB, Johannes E, Rolf MZ. Medical Microbiology, Thieme. 2005;175-180.
Atlas RM. "Principles of Microbiology". 1st ed., Mos by-year Book, Inc., St. Louis, U.S.A.; 1995.
Jeremy WD, Simon FP. Molecular genetics of bacteria 4th ed. John Wiley and Sons Ltd. 2004;137-174.
Strohmair H, Noiges R, Kotschhan S, Swers G, Hogenaner G, Zechner EL, Koraiman G. Sgnal transduction and bacterial conjugation: Characterization of arch A in regulating conjugative transfer of the resistene plasmid R1. J. Mol. Biol. 1998;277:309-316.
Willetts NS, Maule J, Mclntire S. The genetic location of trao pand Tra–4 on the E. coli K12 sex factor F. Genet. Res. 1975; 26(3):255-263.
Brasch MA, Meyer RJ. Genetic orgnization of plasmid R1162 DNA Involved in conjugation mobilization. J. Bacteriol. 1986;167(2):703-710.
Chinedum Ch. S, Abdullah N, Siang TW, Wan HYJ. Plasmid profiling and curing of Lactobacillus strains isolated from gastrointestinal tract of chicken. J. Microbiol. 2005;43(3):251-256.
Hardy K. Bacterial plasmids. Second edition, American Society of Microbiology; 1989.
Kelly AC, Shewmaker FP, Kryndushkin D, Winkner RB. Sex, prions, and plasmids in yeast. PNAS Early Edition. 2012;78:177-207.
Soofy BYN. Diagnosis and Molecular genetic study for yeast of genus Saccharomyces isolated from different sources in Mosul city. Ph.D. Thesis. College of Education for Pure Sciences. University of Mosul. Iraq; 2013.
Malla-Obaida BA, Sultan RH, Jirjees RQ. Detection of positions of genes of β-carotene production by R. mucilaginosa BA61. Rafidain Journal of science. The Third Scientific Conference for Biological Science. 2018;27(5):42-52.
Pan LX, Yang DF, Shao L, Li W, Chen GG, Liang ZQ. Isolation of the oleaginous yeasts from the soil and studies of their lipid-producing capacities. Food Technol. Biotechnol. 2009;47(2):215-220.
Kurtzman CP, Fell JW. The yeasts, A taxonomic study. 4th ed., Elsevier Science B.V., The Netherland; 1998.
Pitt JI, Hocking AD. Fungi and food spoilage. Springer Dordrecht Heidelbery London New York; 2009.
Macfaddin JFM. Biochemical test for identification of medical bacteria. Williams and Willkins. Baltimore. USA; 1985.
Atlas RM, Brown AE, Parks LC. Laboratory Manual of Experimental Microbiology, Mosby Comp; 1995.
Koneman EW, Allen SD, Janda WM, Schreckeberger PC, Win WC. Color atlas and text book of diagnostic microbiology. 5th ed., J. B. Lippincott Raven Publishers, Philadelphia; 1997.
Ernst JF, Chan RK. Characterization of Saccharomyces cerevisiae mutant’s supersensitive to aminoglycoside antibiotics. J. of Bacteriol. 1985;8-14.
Dar N. Isolation and characterization of heavy metals resistant yeast from industrial effluents and their use in environmental cleanup. P h D. Thesis. Punjab. Univ. Lahore. Pkistan; 2004.
Toh-e A, Wickner B. Curing of the 2- μm DNA plasmids from Saccharomyces cerevisiae. J. Bcteriol. 1980;145(3):1421-1424.
Tomoeda M, Inuzka M, Kubo N, Nakamura S. Effective elimination of drug resistance and sex factors in Escherichia coli by sodium dodecyl sulfate. J. Bacteriol. 1968; 95:1078-1089.
Heinemann JA, Sprague JR. Bacterial conjugative plasmids mobilize DNA transfer between bacteria and yeast. Nature. 1989;340.
Heinemann JA, Sprague JR. Transmission of plasmid DNA to yeast by conjugation with bacteria. Methods in Enzymology. 1991;2:187-195.
Delgado JN, Remers WA. Text book of organic medicinal and pharmaceutical chemistry. 10th.ed., a Wolters Kluwer Company. New York; 1998.
Milanezi ACM, Witusk JPD, Vander Sander S. Antifungal susceptibility of yeasts isolated from anthropogenic watershed. Annals of the Brazilian Academy of Sciences. 2019;91(1):2-12.
Wiederhold NP. Antifungal resistance: Current trends and future strategies to combat. Infection and Drug Resistance. 2017;10:249-259.
Hokken MWJ, Zwaan BJ, Melchers WJG, Verweij PE. Facilitators of adaptation and antifungal resistance mechanisms in clinically relevant fungi. Fungal Genetics and Biology. 2019;132 (103254):1-13.
Malla Obaida BAR. Investigate some species of Candida contaminated with yogurt and tested its sensitivity to some antibiotics. Rafidian Journal of Science. 2020;29(2):20-29.
Kawane RS. Studies on antibiotics and heavy metal resistance profiling of Escherichia coli from drinking water and clinical specimens. Bioscience Discovery. 2012;3(3):292-295.
Malla Obaida BAJ. Isolation and identification and study of some Rhodotorula species from genetic and molecular aspects and their efficiency in β-carotene production. ph.D. thesis, College of Education for Pure Sciences, University of Mosul; 2017.
Kawane RS, Tambekar DH. Studies on multidrug and heavy metal resistance in Escherichia coli. 45th Annual conference of Association of Microbiologist of India, NDRL (Karnal). 2004;23-25.
Lucious S, Rrddy ES, Anuradha V, Vijaya PP, Ali MS, Yogananth N, Rajan R, Parveen PK. Heavy metal tolerance and antibiotic sensitivity of bacterial strains isolated from tannery effeluent. Asian J. Exp. Biol. Sci. 2013;4(4):597-606.
Pahwa S, Kaur J, Cameotra SS, Nandanwar H, Kaur P. Curing of multiple plasmids by Et Br in Acinetobacter baumanir. Journal of Advances in Developmental Research. 2012;3(1):82-84.
Jamuna M, Kolanchiammal R, Jeevaratnam K. Plasmid- associated bacteriocin production in Lactobacillus strains isolated from some traditional fermented foods. Global J. Biotech. Biochemi. 2010;5(3): 175-181.
Chigor VN, Umoh VJ, Smith SI, Igbinosa EO, Okoh AI. Multidrug resistance and plasmid patterns of Escherichia coli O157 and Escherichia coli isolated from diarrhoeal stools and surface waters from some selected sources in Zaria, Nigeria. Int. J. Environ. Res. Public Health. 2010;7: 3831-3841.
Daini OA, Adegboyega HO, Odufuwa T, Ogbolu DO. Incidence of multidrug resistance R-plasmids among Escherichia coli causing urinary tract infections: A case study from Nigeria, British Journal of Applied Science and Technology. 2011; 1(4):204-210.
Haslett ND. Evaluating transmission barriers to Escherichia coli x Saccharomyces cerevisiae inter-kingdom conjugation. M. Sc. Thesis. (Hons) in the school of biological Science, Canterbury University; 2006.