Penyimpanan Bakteri Escherichia coli Dan Streptococcus pneumonie Pada Media Cryoprotective Dengan Metode Freeze Drying
Abstract
Proses perbanyakan dan penyimpanan bakteri menjadi hal yang harus diperhatikan karena cara penyimpanan sangat terkait dengan perubahan sifat bakteri yang disimpan. Di Laboratorium, kultur bakteri murni dipindahkan dengan metode subkultur. Kekurangan subkultur berulang adalah memakan waktu, beresiko kontaminasi dan mutasi genetik. Penelitian ini bertujuan untuk mengembangkan penyimpanan bakteri jangka panjang yang aplikatif di Laboratorium Mikrobiologi serta menentukan jenis Cryoprotectan Agent yang dapat mempertahankan viabilitas sel Escherichia coli dan Streptococcus pneumonie dengan metode Freeze Drying. Metode penelitian yang digunakan adalah eksperimen dengan menambahkan Skim Milk 10% dan gliserol 10% sebagai Cryoprotectan Agent pada pelet sel Escherichia coli dan Streptococcus pneumonie dengan metode Freeze Drying. Digunakan kontrol tanpa penambahan Cryoprotectan Agent dengan metode Freeze Drying dan kontrol tanpa penambahan Cryoprotectan Agent yang disimpan dalam refrigator (non Freeze Drying) kemudian jumlah koloni bakteri dihitung setelah masa simpan 2,4,6 minggu.Selanjutnya data yang diperoleh diolah secara statistik menggunakan metode statistik non parametrik kruskal wallis . Hasil penelitian menunjukkan bahwa Skim Milk 10% mampu mempertahankan viabilitas Sel Escherichia coli dengan persentase penurunan viabilitas paling rendah yaitu 4,2%, namun jenis Cryoprotectan agent metode Freeze Dry belum mampu mengawetkan dan mempertahankan viabilitas sel Streptococcus pneumonie.
Kata Kunci: Cryoprotectan Agent; Cell viability; Escherichia coli;, Streptococcus pneumonie; Freeze DryingReferences
A.M. Mitchell, T.J. Mitchell. 2010.Streptococcus pneumoniae: virulence factors and variation, Clinical Microbiology and Infection, Volume 16, Issue 5, Pages 411-418
Abadias et al. 2001. Effect of freeze-drying and protectants on viability of the biocontrol yeast Candida sake. International Journal of Food Microbiology. (65): 173–182.
Acharya, Tankeshwar. 2010. Maintenance and Preservation of Pure Cultures of BacteriaI. [Online].Tersedia:http://microbeonline.com/maintenance-andpreservation-of-pure-cultures-of-bacteria. [15/06/2017].
Alegre, Santiago V, Isabel Fernández-Natal, Eduardo López-Fidalgo, Octavio Miguel Rivero-Lezcano. 2015. Preparation of inocula for experimental infection of blood with Streptococcus pneumoniae, MethodsX, Volume 2, Pages 463-468.
Beatriz Quintero Moreno, María Araque, and Evelyn Mendoza. 2017. “Evaluation of Two Supplemented Culture Media for Long-Term, Room-Temperature Preservation of Streptococcus pneumoniae Strains,” BioMed Research International, vol. 2017, Article ID 1218798, 9 pages. https://doi.org/10.1155/2017/1218798.
Cappuccino & Sherman. 2014. Manual Labortaorium Mikrobiologi. Jakarta : EGC.
Carvalho et al. 2004. Relevant Factors For The Preparation of Freeze-Dried Lactic Acid Bacteria. International Journal. (14) : 835 – 847.
Cody et al. 2008. Skim Milk Enhances the Preservation of Thawed -80°C Bacterial Stocks. J Microbiol Methods Vol. 75(1) : 135-138.
Criste A. et al. 2014. Research Concerning Use of Long-Term Preservation Technique for Microorganism. Scientific Papers: Animal Science and Biotecnologies Vol. 47(2) : 400372.
Jawetz, Melnick & Adelberg. 2014. Mikrobiologi Kedokteran Edisi ke 25. Jakarta : EGC.
Leslie S. et al. 1995. Trehalose and Sucrose Protect Both Membranes and Proteins in Intact Bacteria during Drying. Applied and Environtmental, Microbiology Vol. 61(10) : 0099-2240.
Martin-Dejardin et al. 2013. A Way To Follow The Viability On Encapsulated Bifidobacterium Bifidium Subjected To A Freeze – Drying Process In Target The Colon : Interest Of Flow Cytometry. Microbiology Review Vol.13(3): 470-511.
Parveen Jamal et al. 2016. Bacterial Preservation for Short and Long-term Stroage. Experimental Methods in Modern Biotechnology. Vol. 2 (12):103.
Ranjan, S., Bakthavathsalam, G., Raghavan, V. V., & Sindhu, R. K. 2014. Growth of Streptococcus pneumoniae on Macconkey Agar: Possibility of Impossible?. Journal of clinical and diagnostic research : JCDR, 8(12), DL01.doi:10.7860/JCDR/2014/10882.5328
S. Kandil & M. El Soda. 2015. Influence of Frezee Drying on Intracellular Enzymatic Activity and Autolytic Properties of Some Lactic Acid Bacterial Strain. Advances in Microbiology (5) : 371-382.
Selwal, Krishan & K. Selwal, Manjit & Gandhi, D.N.. (2011). Effect of freeze drying process on some properties of Streptococcus thermophilus isolated from dairy products. Brazilian Journal of Microbiology. 42. 10.1590/S1517-83822011000400037.
Shukla, S. 2011. Freeze Drying Process : A Review Article. International Journal of Pharmaceutical Sciences and Research Vol. 2 (12) : 3061-3068.
Siberry, G., Brahmadathan, K. N., Pandian, R., Lalitha, M. K., Steinhoff, M. C., & John, T. J. (2001). Comparison of different culture media and storage temperatures for the long-term preservation of Streptococcus pneumoniae in the tropics. Bulletin of the World Health Organization, 79(1), 43–47.
Susilawati & Purnomo. 2016. Viabilitas Sel Bakteri Dengan Cryoprotectant Agent Berbeda (Sebagai Acuan Dalam Preservasi Culture Collection di Laboratorium Mikrobiologi). Biogenesis Jurnal Ilmiah Biologi Vol. 4 (1) : 2302-1616.
Wasas, Avril D. , Robin E. Huebner, Mignon De Blanche, Keith P. Klugman. 1998. Long-Term Survival of Streptococcus pneumoniae at Room Temperature on Dorset Egg Medium Journal of Clinical Microbiology, 36 (4) 1139-1140.
Copyright (c) 2020 Quagga: Jurnal Pendidikan dan Biologi
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
