Effect of Growth Conditions on Hemolysin Production from Clinical Isolates of Escherichia Coli
DOI:
https://doi.org/10.23851/mjs.v35i3.1442Keywords:
E. coli, Hemolysin, Growth conditions, Clinical specimens, Normal floraAbstract
Background: Escherichia coli belongs to the Enterobacteriaceae family and is found in the digestive systems of humans and various animals as normal flora. Objective: This study aims to identify the optimal conditions necessary for the synthesis of hemolysin by E. coli isolated from a variety of clinical contexts. Methods: A total of 120 specimens were collected from different clinical sources in Baghdad Hospitals. Screening for hemolysin production was done by plate and Spectrophotometric methods. Furthermore, many conditions for producing hemolysin from clinical E. coli isolates were investigated including temperature, pH, incubation time, inoculum size, culture medium, and aeration. Results: Six isolates of E. coli exhibited hemolysin production. Optimal conditions for hemolysin production were determined: the temperature was 37 °C with an optical density of 0.931, the pH was 7 with an optical density of 0.976, the incubation period was 24 hours with an optical density of 0.885, 1% inoculum resulting in an optical density of 0.558, brain heart infusion as the best medium with an optical density of 1.697, and a shaker method showing superior results (optical density of 1.235) compared to static aeration (optical density of 0.988). Conclusions: This study identified the optimal conditions necessary for the synthesis of hemolysin by E. coli. By understanding these optimal conditions, researchers may be better equipped to develop strategies for managing and controlling the spread of these pathogenic bacteria.
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References
I. Naqid, A. Balatay, N. Hussein, K. Saeed, H. Ahmed, and S. Yousif, "Antibiotic susceptibility pattern of escherichia coli isolated from various clinical specimens in duhok city, kurdistan region of iraq," International Journal of Infection, vol. 7, no. 3, pp. 1-6, 2020.
C. Schindel, A. Zitzer, B. Schulte, A. Gerhards, P. Stanley, C. Hughes, V. Koronakis, S. Bhakdi, and M. Palmer, "Interaction of e. coli hemolysin with biological membrane," Journal of Biochemistry, vol. 268, no. 3, pp. 800-808, 2004.
J. Chester and D. Kyung, "Bacterial pore-forming hemolysins and their use in the cytosolic delivery of macromolecules," Advanced Drug Delivery Reviews, vol. 41, pp. 209-221, 2000.
K. Murase, T. Ooka, A. Iguchi, Y. Ogura, K. Nakayama, M. Asadulghani, M. Islam, H. Hiyoshi, T. Kodama, L. Beutin, and T. Hayashi, "Haemolysin e- and enterohaemolysin-derived haemolytic activity of 055/0157 strains and other escherichia coli lineages," Microbiology, vol. 158, no. 3, pp. 746-758, 2012.
Y. Burgos and L. Beutin, "Common origin of plasmid-encoded alpha-hemolysin genes in escherichia coli," BMC Microbiology, vol. 10, no. 1, pp. 1-13, 2010.
M. Bielaszewska, C. Rüter, L. Kunsmann, L. Greune, A. Bauwens, W. Zhang, T. Kuczius, K. S. Kim, A. Mellmann, M. A. Schmidt, and H. Karch, "Enterohemorrhagic escherichia coli hemolysin employs outer membrane vesicles to target mitochondria and cause endothelial and epithelial apoptosis," PLoS Pathogens, vol. 9, no. 12, e1003797, 2013.
K. Kanonenberg, O. Spitz, I. Erenburg, T. Beer, and L. Schmitt, "Type i secretion system-it takes three and a substrate," FEMS Microbiology Letters, vol. 365, no. 11, fny094, 2018.
B. D. L. Caetano, M. d. O. Domingos, M. A. da Silva, J. C. A. da Silva, J. M. Polatto, F. Montoni, L. K. Iwai, D. C. Pimenta, H. Vigerelli, P. C. G. Vieira, R. d. C. Ruiz, J. S. Patané, and R. M. F. Piazza, "In silico prediction and design of uropathogenic escherichia coli alpha-hemolysin generate a soluble and hemolytic recombinant toxin," Microorganisms, vol. 10, no. 1, p. 172, 2022.
P. Tille, Bailey & Scott's Diagnostic Microbiology, 13th. St. Louis, Missouri: Mosby Elsevier Publications, 2014, pp. 484-512.
A. Brown and H. Smith, Benson's Microbiological Applications Laboratory Manual in General Microbiology, 13th. USA: McGraw-Hill Education, 2015.
G. Di Venanzio, T. M. Stepanenko, and E. Garcia Vescovi, "Serratia marcescens shla pore-forming toxin is responsible for early induction of autophagy in host cells and is transcriptionally regulated by rcsb," Infection and Immunity, vol. 82, no. 9, pp. 3542-3554, 2014.
M. Akram, M. Shahid, and A. Khan, "Etiology and antibiotic resistance patterns of community-acquired urinary tract infections in jnmc hospital aligarh, india," Annals of Clinical Microbiology and Antimicrobials, vol. 6, no. 1, pp. 1-7, 2007.
H. Ghaffoori and A. Suleiman, "Isolation, identification, and antibiotic resistance profile distribution of clinical e. coli in iraqi patients," Eurasian Medical Research Periodical, vol. 8, pp. 116-121, 2022.
R. Kadhim Mohammed, "Investigation of the role of virulence gene in biofilm formation of escherichia coli obtained from clinical specimens in baghdad," Archives of Razi Institute, vol. 77, no. 2, pp. 915-921, 2022.
A. Al-Sabawi and D. Jwher, "A comparative study of escherichia coli isolates from open and closed sheep breeding systems in nineveh, iraq," Basrah Journal of Veterinary Research, vol. 21, no. 3, pp. 12-22, 2022.
A. Rhumaid, A. Al-Tabtabai, J. Al-Buhilal, and N. Al-Rubaey, "Detection and characterization of fimh gene in escherichia coli isolated from urine specimens of diabetic adult females with urinary tract infection," Annals of the Romanian Society for Cell Biology, pp. 13 948-13 953, 2021.
A. Mahe, B. Sabiu, A. Adam, and U. Abdullahi, "Effect of citric acid at different ph on the survival of escherichia coli," Bayero Journal of Pure and Applied Sciences, vol. 14, no. 1, pp. 79-84, 2021.
M. Bagatini, C. Assmann, and M. Blumenberg, Glutathione System and Oxidative Stress in Health and Disease. BoD-Books on Demand. IntechOpen, 2020.
M. Madigan and J. Martinko, Brock Biology of Microorganisms, 12th. Upper Saddle River, NJ: Benjamin Cum mings, 2008.
A. Hussein and A. Al-Hasany, "Isolation and diagnosis of providencia rettgeri from different clinical cases and study antibiotics resistant," Al-Qadisiyah Journal of Pure Science, vol. 22, no. 3, pp. 109-120, 2017.
A. Mare, A. Man, F. Toma, C. N. Ciurea, R. L. Coseriu, C. Vintilă, and A. C. Maier, "Hemolysin-producing strains among diarrheagenic escherichia coli isolated from children under 2 years old with diarrheal disease," Pathogens, vol. 9, no. 12, pp. 1-12, 2020.
E. Bı́rošová, L. Siegfried, M. Kmet’ova, A. Makara, A. Ostró, A. Grešová, P. Urdzı́k, A. Liptáková, M. Molokáčová, R. Bártl, and L. Valansky, "Detection of virulence factors in I+--haemolytic escherichia coli strains isolated from various clinical materials," Clinical Microbiology and Infection, vol. 10, no. 6, pp. 569-573, 2004.
M. Rizvi, S. Kumar, F. Khan, A. Sultan, A. Malik, and F. Tahira, "Killing of alpha-hemolytic and non-hemolytic escherichia coli strains from paediatric patients in human serum and polymorphonuclear leucocytes," International Journal of Current Microbiology and Applied Sciences, vol. 2, no. 11, pp. 636-646, 2013.
R. Vaish, M. Pradeep, C. Setty, and V. Kandi, "Evaluation of virulence factors and antibiotic sensitivity pattern of escherichia coli isolated from extraintestinal infections," Cureus, vol. 8, no. 5, pp. 1-13, 2016.
N. Srikanth and R. Macaden, "Uropathogenic escherichia coli: A preliminary study," Indian Journal of Pathology and Microbiology, vol. 46, pp. 6-145, 2003.
A. Valeva, I. Walev, H. Kemmer, S. Weis, I. Siegel, F. Boukhallouk, T. M. Wassenaar, T. Chavakis, and S. Bhakdi, "Binding of e. coli hemolysin and activation of target cell is not receptor dependent," Journal of Biological Chemistry, vol. 280, no. 44, pp. 36 657-36 663, 2005.
H. Al-Shammary, S. Al-Hasani, and M. Nadir, "Extraction and studying the effect of pH and temperature on hemolysin production by a local isolate of staphylococcus aureus," Kerbala Journal of Medicine, vol. 5, no. 11, pp. 1283-1294, 2012.
M. Razzaq, I. Bnyan, and H. Al-Hasson, "Extraction and immobilization of hemolysin a produced by uropathogenic escherichia coli," Medical Journal of Babylon, vol. 8, no. 2, pp. 157-163, 2011.
M. Naskar, V. P. Parekh, M. A. Abraham, Z. Alibasic, M. J. Kim, G. Suk, J. H. Noh, K. Y. Ko, J. Lee, C. Kim, H. Yoon, S. N. Abraham, and H. W. Choi, "α-hemolysin promotes uropathogenic e. coli persistence in bladder epithelial cells via abrogating bacteria-harboring lysosome acidification," PLoS Pathogens, vol. 19, no. 5, pp. 1-29, 2023.
Y. Qu, R. Zhang, F. Ma, J. Zhou, and B. Yan, "Bioaugmentation with a novel alkali-tolerant pseudomonas strain for alkaline phenol wastewater treatment in sequencing batch reactor," World Journal of Microbiology and Biotechnology, vol. 27, no. 8, pp. 1919-1926, 2011.
Z. Yang and F. Wu, "Catalytic properties of tyrosinase from potato and edible fungi," Biotechnology, vol. 5, no. 3, pp. 344-348, 2006.
B. Turner, "Variation in ph optima of hydrolytic enzyme activities in tropical rainforest soils," Applied and Environmental Microbiology, vol. 76, no. 19, pp. 6485-6493, 2010.
A. Abdulwahhab and K. Khalaf, "Effect of cultivation conditions on hemolysin production from clinical isolates of serratia marcescens," Al-Mustansiriyah Journal of Science, vol. 33, no. 1, pp. 6-14, 2022.
N. Maarib, H. Mohaisen, and J. AL-K., "Isolation, molecular identification, and influence of incubation period on hemolysin gene expression in serratia marcescens local isolates," International Journal of Natural and Engineering Sciences, vol. 14, no. 1, pp. 3-7, 2020.
S. Stonsaovapak, "Effects of temperature and cell number on the production of hemolysin by vibrio parahaemolyti cus," Agriculture and Natural Resources, vol. 34, no. 2, pp. 248-253, 2000.
S. Mabrouk, A. Hashem, N. El-Shayeb, A. Ismail, and A. Abdel-Fattah, "Optimization of alkaline protease pro ductivity by bacillus licheniformis atcc 21415," Bioresource Technology, vol. 69, no. 2, pp. 155-159, 1999.
R. Abd Rahman, L. Geok, M. Basri, and A. Salleh, "Physical factors affecting the production of organic solvent tolerant protease by pseudomonas aeruginosa strain k," Bioresource Technology, vol. 96, no. 4, pp. 429-436, 2005.
Z. Rahim and K. Aziz, "Factors affecting production of haemolysin by strains of vibrio fluvialis," Journal of Diarrhoeal Diseases Research, pp. 113-116, 1996.
F. Mantzouridou, T. Roukas, and P. Kotzekidou, "Effect of the aeration rate and agitation speed on β-carotene production and morphology of blakeslea trispora in a stirred tank reactor: Mathematical modeling," Biochemical Engineering Journal, vol. 10, no. 2, pp. 123-135, 2002.
A. Kamble, V. Meena, and U. Banerjee, "Effect of agitation and aeration on the production of nitrile hydratase by rhodococcus erythropolis mtcc 1526 in a stirred tank reactor," Letters in Applied Microbiology, vol. 51, no. 4, pp. 413-420, 2010.
T. Zotta, E. Parente, and A. Ricciardi, "Aerobic metabolism in the genus lactobacillus: Impact on stress response and potential applications in the food industry," Journal of Applied Microbiology, vol. 122, no. 4, pp. 857-869,2017.
E. Elmansy, M. Asker, E. El-Kady, S. Hassanein, and F. El-Beih, "Production and optimization of α-amylase from thermo-halophilic bacteria isolated from different local marine environments," Bulletin of the National Research Centre, vol. 42, no. 1, pp. 1-9, 2018.
R. Benedict and F. Schultz, "An effect of iron level and food type on production of hemolysin and catalase of listeria monocytogenes strain scott a," Journal of Food Protection, vol. 54, no. 7, pp. 528-531, 1991.
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