The Potential Use of EDTA as an Alternative to Defibrination in Preparing Blood Agar Plates with Human AB Blood Type on Staphylococcus aureus Culture
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https://orcid.org/0000-0002-8560-4510
Eka Puspitasari Fanny Kurnanda Razvi
Abstract
Blood Agar Plates (BAP) are composed of blood as one of the compositions. Sheep’s blood is usually used, but since it is difficult to be obtained, human AB blood type was used as an alternative. In preparing BAP, blood is defibrinated to lyse the blood clotting factors. Blood clots can also be prevented by adding anticoagulants, such as ethylenediaminetetraacetic acid (EDTA). This study aims to investigate the potential use of EDTA as a substitute for defibrination in preparing BAP with human AB blood type. This study employed a completely randomized design with true experimental method using Staphylococcus aureus as the sample. The parameters were the number of colonies, types of hemolysis, and hemolysis zone. The results showed that the S. aureus grown on BAP with EDTA-human AB blood type was 64 colonies (mean), produced β-hemolytic pattern, and 6 mm hemolytic zone. In contrast, the S. aureus grown on BAP with defibrinated human AB blood type showed 82 colonies (mean), β-hemolytic pattern, and 5 mm hemolytic zone. There were significant differences in the number of colonies (0.000 < α) and hemolytic zones (0.02 < α). However, there was no difference in the hemolysis type (both treatments produced β-hemolysis). EDTA was possible to be used as a substitute for defibrination in preparing BAP to assess the hemolysis type of S. aureus, but it might not be able to be used as a benchmark for counting the number of colonies and determining the hemolysis zone of S. aureus.
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Copyright (c) 2021 Dora Dayu Rahma Turista, Eka Puspitasari, Fanny Kurnanda Razvi
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References
Turista DDR., Puspitasari E. The Growth of Staphylococcus aureus in the blood agar plate media of sheep blood and human blood groups A, B, AB, and O. J Teknol Lab. 2019; 8(1): 1-7.
Russell FM, Biribo SSN, Selvaraj G, Oppedisano F, Warren S, Seduadua A, et al. As a bacterial culture medium, citrated sheep blood agar is a practical alternative to citrated human blood agar in laboratories of developing countries. J Clin Microbiol. 2006; 44(9): 3346-51.
Shabnam I, S CD, C JB. Ethylenediaminetetraacetic acid (EDTA) - Dependent pseudothrombocytopenia: a case report. J Clin Diagnostic Res. 2014; 8(10):
Yeh E, Pinsky BA, Banaei N, Baron EJ. Hair sheep blood, citrated or defibrinated, fulfills all requirements of blood agar for diagnostic microbiology laboratory tests. PLoS One. 2009; 4(7).
Moraveji Z, Tabatabaei M, Shirzad Aski H, Khoshbakht R. Characterization of hemolysins of Staphylococcus strains isolated from human and bovine, southern Iran. Iran J Vet Res. 2014; 15(4): 326-30.
Silva ER da, Boechat JUD, Martins JCD, Ferreira WPB, Siqueira AP, Silva N da. Hemolysin production by Staphylococcus aureus species isolated from mastitic goat milk in Brazilian dairy herds. Small Rumin Res [Internet]. 2005; 56(1-3): 271-5.
Bonnet M, Lagier JC, Raoult D, Khelaifia S. Bacterial culture through selective and non-selective conditions: the evolution of culture media in clinical microbiology. New Microbes New Infect. 2020; 34.
Melani D, Radiati LE, Thohari I. The addition Of EDTA (ethylenediaminetetraacetic acid) with egg white lysozyme extracts as the antimicrobial activity on Salmonella sp and Staphylococcus. 2013; 1-8.
Khan A, Vu KD, Riedl B, Lacroix M. Optimization of the antimicrobial activity of nisin, Na-EDTA and pH against gram-negative and gram-positive bacteria. LWT - Food Sci Technol [Internet]. 2015; 61(1): 124-9.
Farca AM, Nebbia P, Re G. Potentiation of antibiotic activity by EDTA-tromethamine against three clinically isolated Gram-positive resistant bacteria. Anin vitro investigation. Vet Res Commun [Internet]. 1994; 18(1): 1-6.
Sohlenkamp C, Geiger O. Bacterial membrane lipids: diversity in structures and pathways. Narberhaus F, editor. FEMS Microbiol Rev [Internet]. 2016; 40(1): 133-59.
Colobert L. [Inactivation by lysozyme of the somatic antigen of heated pathogenic Salmonella]. C R Hebd Seances Acad Sci. 1957;244(23): 2863-5.
Noller EC, Hartsell SE. Bacteriolysis of Enterobacteriaceae. I. Lysis by four lytic systems utilizing lysozyme. J Bacteriol [Internet]. 1961; 81(3): 482-91.
Noller EC, Hartsell SE. Bacteriolysis of Enterobacteriaceae. II. Pre- and co-lytic treatments potentiating the action of lysozyme. J Bacteriol [Internet]. 1961; 81(3): 492-9.
Gray GW, Wilkinson SG. The effect of ethylenediaminetetra-acetic acid on the cell walls of some gram-negative bacteria. J Gen Microbiol [Internet]. 1965; 39(3): 385-99.
Oktari A, Vanawati N, Kurniawati UI. The optimization of Human Blood Agar (HBA) for Streptococcus pneumonia growth. J Phys Conf Ser. 2019; 1280(2): 3-9.
Buxton R. Blood Agar Plates and Hemolysis Protocols. In: American Society for Microbiology. 2016. p. 1-9.
Makhro A, Huisjes R, Verhagen LP, Mañú-Pereira M del M, Llaudet-Planas E, Petkova-Kirova P, et al. Red cell properties after different modes of blood transportation. Front Physiol. 2016; 7(JUL): 1-21.
Huseby M, Shi K, Kent Brown C, Digre J, Mengistu F, Keun SS, et al. Structure and biological activities of beta toxin from Staphylococcus aureus. J Bacteriol. 2007; 189(23): 8719-26.
Dinges MM, Orwin PM, Schlievert PM. Exotoxins of Staphylococcus aureus. Clin Microbiol Rev. 2000; 13(1): 16-34.
Löffler B, Hussain M, Grundmeier M, Brück M, Holzinger D, Varga G, et al. Staphylococcus aureus panton-valentine leukocidin is a very potent cytotoxic factor for human neutrophils. PLoS Pathog. 2010; 6(1): 1-12.
Laham N Al, Mediavilla JR, Chen L, Abdelateef N, Elamreen FA, Ginocchio CC, et al. MRSA clonal complex 22 strains harboring toxic shock syndrome toxin (TSST-1) are endemic in the primary hospital in Gaza, Palestine. PLoS One. 2015; 10(3): 1-17.
