Viability of Gram-Positive and Gram-Negative Bacteria in Liquid Storage Media

Tacik Idayanti Idayanti; Musholli Himmatun Nabilah Nabilah; Retno Werdiningsih

Penulis

Tacik Idayanti Idayanti
Poltekkes Kemenkes Surabaya, Surabaya, Indonesia
Musholli Himmatun Nabilah Nabilah
Poltekkes Kemenkes Surabaya, Surabaya, Indonesia
Retno Werdiningsih
Poltekkes Kemenkes Surabaya, Surabaya, Indonesia

Kata Kunci:

Bouillon, Gram-positive, Gram-negative, Tryptic Soy Broth

Abstrak

Bacteria can be cultivated and stored in media. In educational laboratories, liquid media are used to maintain stock cultures due to simplicity and cost-effectiveness. Maintaining bacterial viability is essential to ensure bacterial stability. Several factors influence bacterial survival, including temperature storage, duration, nutrient composition, osmotic pressure, pH, and sterility. Previous studies have applied glycerol supplementation in Tryptic Soy Broth as a preservative to improve bacterial viability. This study to observe bacterial survival in two types of liquid Bouillon and Tryptic Soy Broth (TSB)—with storage durations of one week, two weeks, and one month. The spread plate method used "L" glass rod on Nutrient Agar, with four treatment groups and replications. The results showed a significant increase numbers of bacteria in TSB after two weeks of storage, followed by a sharp decrease after one month. Showed that TSB was better at maintaining the viability of Gram-positive and Gram-negative bacteria.

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Biografi Penulis

Tacik Idayanti Idayanti, Poltekkes Kemenkes Surabaya, Surabaya, Indonesia

Department of Medical Laboratory Technology, Poltekkes Kemenkes Surabaya
Musholli Himmatun Nabilah Nabilah, Poltekkes Kemenkes Surabaya, Surabaya, Indonesia

Musholli Himmatun Nabilah, S.Tr. Kes is a medical laboratory technician at the Department of Medical Laboratory Technology, Poltekkes Kemenkes Surabaya
Retno Werdiningsih, Poltekkes Kemenkes Surabaya, Surabaya, Indonesia

Retno Werdiningsih, S.ST. is a medical laboratory technician at the Department of Medical Laboratory Technology, Poltekkes Kemenkes Surabaya

Referensi

[1] D. T. Afriani, A. Rofieq, A. M. Huda, and R. Wahyu, “Analysis Of Microbial Culture Learning : Understanding The Relationship Between Technical Competence And Students ’ Perspectives At Dharmawangsa University , Medan,” vol. 10, no. March, pp. 207–218, 2024.

[2] A. M. Arista, E. A. Rizquita, and R. Y. Oxi, “Effectiveness of Bacterial Culture Preservative Techniques in the Microbiology Laboratory,” vol. 1, no. 1, pp. 1–6, 2023.

[3] G. H. Peralta et al., “Impact of media culture, freeze-drying and storage conditions on preservation of Lacticaseibacillus paracasei 90: Viability and metabolic potential as a secondary culture in semi-hard cheese,” Int. Dairy J., vol. 147, p. 105763, 2023, doi: https://doi.org/10.1016/j.idairyj.2023.105763.

[4] O. Dimciu, “The temperatures influence on the development of Staphylococcus genus bacteria.” 2007.

[5] A. Wanger, V. Chavez, R. S. P. Huang, A. Wahed, J. K. Actor, and A. Dasgupta, “Media for the Clinical Microbiology Laboratory,” Microbiol. Mol. Diagnosis Pathol., pp. 51–60, 2017, doi: 10.1016/b978-0-12-805351-5.00004-1.

[6] K. Rahma, J. C. Warella, S. Artikel, and K. Kunci, “Pertumbuhan Biofilm Escherichia Coli Pada Media Tryptic Soy Broth,” J. Ilm. Sain dan Teknol., vol. 2, no. 4, pp. 164–169, 2024.

[7] L. Walker, S. Sun, and H. Thippareddi, “The Minimum Growth Temperatures of Shiga Toxin–Producing Escherichia coli,” Foodborne Pathog. Dis., Mar. 2025, doi: 10.1089/fpd.2024.0108.

[8] L.-M. Sittek, J. Hunold, F. E. Runkel, and P. Schlupp, “Comprehensive analysis of Staphylococcus aureus supernatants in different culture media to mimic chronic wound conditions in vitro,” The Microbe, vol. 4, no. September, p. 100161, 2024, doi: 10.1016/j.microb.2024.100161.

[9] G. Wijesinghe, A. Dilhari, B. Gayani, N. Kottegoda, L. Samaranayake, and M. Weerasekera, “Influence of Laboratory Culture Media on in vitro Growth, Adhesion, and Biofilm Formation of Pseudomonas aeruginosa and Staphylococcus aureus,” Med. Princ. Pract., vol. 28, no. 1, pp. 28–35, 2019, doi: 10.1159/000494757.

[10] M. Ishii, Y. Matsumoto, and K. Sekimizu, “Compounds in a particular production lot of tryptic soy broth inhibit Staphylococcus aureus cell growth,” Drug Discov. Ther., vol. 9, no. 3, pp. 178–183, 2015, doi: 10.5582/ddt.2015.01030.

[11] N. P. I. Lestari and A. A. A. P. Permatasari, “Pengaruh Suhu Dan Waktu Simpan Terhadap Populasi Total Bakteri, Coliform Dan Escherichia coli Pada Ikan Nila (Oreochromis niloticus),” J. Media Sains, vol. 2, no. 2, pp. 96–103, 2018, doi: 10.36002/jms.v2i2.429.

[12] V. Agustiarini and D. Permata Wijaya, “Pengaruh waktu penyimpanan inokulum Escherichia coli dan Staphilococcus aureus pada suhu dingin terhadap jumlah sel bakteri di Laboratorium Mikrobiologi,” J. Penelit. Sains, vol. 21, no. 3, pp. 117–124, 2021, [Online]. Available: http://ejurnal.mipa.unsri.ac.id/index.php/jps/index.

[13] E. Vasilakou, M. C. M. Van Loosdrecht, and S. A. Wahl, “Escherichia coli metabolism under short-Term repetitive substrate dynamics: Adaptation and trade-offs,” Microb. Cell Fact., vol. 19, no. 1, pp. 1–19, 2020, doi: 10.1186/s12934-020-01379-0.

[14] Sunarno et al., “Long-term Storage of Bacterial Isolates by Using Tryptic Soy Broth with 15% Glycerol in the Deep Freezer (-70 to -80 C),” IOP Conf. Ser. Earth Environ. Sci., vol. 913, no. 1, 2021, doi: 10.1088/1755-1315/913/1/012070.

[15] C. A. Bardsley, M. J. Young, M. Sharma, C. Kessler, C. B. Appolon, and K. R. Schneider, “Growth Media of Escherichia coli Does Not Affect Its Survival in Soil under Static Conditions,” J. Food Prot., vol. 85, no. 12, pp. 1842–1847, 2022, doi: 10.4315/JFP-22-082.

[16] A. Hasan and P. R. Wikandari, “Penentuan waktu produksi optimum bakteriosin asal Lactobacillus plantarum B1765 berdasarkan aktivitas penghambatannya terhadap Staphylococcus aureus,” J. Chem., vol. 7, no. 1, pp. 15–20, 2018.

[17] F. Petersen and J. A. Hubbart, “Physical factors impacting the survival and occurrence of escherichia coli in secondary habitats,” Water (Switzerland), vol. 12, no. 6, pp. 1–15, 2020, doi: 10.3390/w12061796.

[18] A. R. Tuttle, N. D. Trahan, and M. S. Son, “Growth and Maintenance of Escherichia coli Laboratory Strains,” Curr. Protoc., vol. 1, no. 1, pp. 1–13, 2021, doi: 10.1002/cpz1.20.

[19] C. O. Gill, M. Badoni, and T. H. Jones, “Behaviours of log phase cultures of eight strains of Escherichia coli incubated at temperatures of 2, 6, 8 and 10 °C,” Int. J. Food Microbiol., vol. 119, no. 3, pp. 200–206, 2007, doi: 10.1016/j.ijfoodmicro.2007.07.043.

[20] Madigan, Bender, Buckley, Sattley, and Stahl, Brock Biology of Microorgisme, no. May. 2018.

[21] J. M. Willey, L. M. Prescott, K. M. Sandman, and D. H. (Dorothy H. Wood, Cap 3 Prescott’s microbiology. 2023.

[22] A. Visciglia et al., “Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies,” Front. Microbiol., vol. 13, no. November, pp. 1–8, 2022, doi: 10.3389/fmicb.2022.989563.

[23] J. E. Doyle, W. H. Mehrhof, and R. R. Ernst, “Limitations of thioglycolate broth as a sterility test medium for materials exposed to gaseous ethylene oxide.,” Appl. Microbiol., vol. 16, no. 11, pp. 1742–1744, 1968, doi: 10.1128/aem.16.11.1742-1744.1968.

[24] E. N. Mahmmoud, “Comparison of preservation methods of staphylococcus aureus and escherichia coli bacteria,” J. Pure Appl. Microbiol., vol. 14, no. 3, pp. 2173–2180, 2020, doi: 10.22207/JPAM.14.3.58.