GAMICBANK: Pendekatan Inovatif dalam Pelestarian Isolat Mikroba
Mulyani Mulyani(1*), Kamla Awaludin(2), Ayu Rahayu(3), Titik Nuryastuti(4)
(1) Departemen Mikrobiologi, Fakultas Kedokteran, Kesehatan Masyarakat, dan Keperawatan, Universitas Gadjah Mada, Yogyakarta, Indonesia
(2) Departemen Mikrobiologi, Fakultas Kedokteran, Kesehatan Masyarakat, dan Keperawatan, Universitas Gadjah Mada, Yogyakarta, Indonesia
(3) Departemen Mikrobiologi, Fakultas Kedokteran, Kesehatan Masyarakat, dan Keperawatan, Universitas Gadjah Mada, Yogyakarta, Indonesia
(4) Departemen Mikrobiologi, Fakultas Kedokteran, Kesehatan Masyarakat, dan Keperawatan, Universitas Gadjah Mada, Yogyakarta, Indonesia
(*) Corresponding Author
Abstract
ABSTRAK. Pelestarian isolat mikroba sangat penting untuk penelitian, diagnostik, dan bioteknologi. Metode konvensional seperti stok gliserol atau liofilisasi efektif namun seringkali mahal dan rentan terhadap risiko kontaminasi. Strategi alternatif penyimpanan berbasis butiran, memungkinkan pengambilan mikroorganisme yang masih hidup dengan mudah sambil meminimalkan siklus pembekuan-pencairan berulang. Studi ini bertujuan untuk mengevaluasi kinerja awal sistem GAMICBANK, metode penyimpanan mikroba berbasis butiran menggunakan butiran mikrobank plastik yang dimasukkan dalam larutan susu skim atau Skim Milk–Tryptone–Glucose–Glycerol (STGG), untuk menyimpan bakteri dan ragi pada suhu –80 °C. Isolat Gram-positif, Gram-negatif, dan ragi yang representatif (Staphylococcus aureus, Streptococcus spp., Listeria monocytogenes, Salmonella typhi, Shigella flexneri, Escherichia coli, Neisseria gonorrhoeae, Chromobacterium violaceum, dan Candida albicans) disimpan dalam butiran dengan susu skim atau STGG. Butiran tersebut diambil kembali setelah 1 dan 3 bulan, ditanam pada media selektif, dan dievaluasi pertumbuhan koloni dan viabilitasnya. Setelah 1 bulan, semua isolat tetap memiliki viabilitas yang sangat baik di kedua media. Pada 3 bulan, sebagian besar spesies tetap sangat viabel (>99%) di kedua media. Namun, Listeria monocytogenes dan Neisseria gonorrhoeae menunjukkan penurunan viabilitas yang signifikan dalam susu skim (30%) tetapi tetap stabil dalam STGG (99%). Demikian pula, Candida albicans menunjukkan penurunan viabilitas dalam susu skim (50%) tetapi mempertahankan 99% kelangsungan hidup dalam STGG. Sistem bead GAMICBANK secara efektif melestarikan mikroorganisme yang tangguh dan sensitif pada suhu –80 °C. STGG memberikan perlindungan beku yang lebih unggul dibandingkan susu skim, terutama untuk bakteri sensitif dan ragi. Kesimpulannya bahwa sistem GAMICBANK dapat dijadikan sebagai metode alternatif untuk penyimpanan isolat bakteri dan ragi.
ABSTRACK.The preservation of microbial isolates is crucial for research, diagnostics, and biotechnology. Conventional methods, such as glycerol stocks or lyophilization, are effective but frequently expensive and susceptible to contamination risks. An alternative strategy, bead-based storage, facilitates the easy retrieval of viable microorganisms while minimizing repeated freeze-thaw cycles. This study aims to evaluate the initial performance of the GAMICBANK system, a bead-based microbial storage method utilizing plastic microbank beads submerged in either skim milk or Skim Milk–Tryptone–Glucose–Glycerol (STGG) solutions, for the cryopreservation of bacteria and yeast at –80 °C. Representative Gram-positive, Gram-negative, and yeast isolates (Staphylococcus aureus, Streptococcus spp., Listeria monocytogenes, Salmonella typhi, Shigella flexneri, Escherichia coli, Neisseria gonorrhoeae, Chromobacterium violaceum, and Candida albicans) were preserved in beads using either skim milk or STGG. The beads were retrieved after 1 and 3 months of storage, cultured on selective media, and evaluated for colony growth and viability. After 1 month, all isolates maintained excellent viability in both preservation media. At 3 months, the majority of the species remained highly viable (>99%) in both media. However, Listeria monocytogenes and Neisseria gonorrhoeae exhibited a significant decline in viability when stored in skim milk (30%) but remained stable in STGG (99%). Similarly, Candida albicans demonstrated a reduction in viability in skim milk (50%) while maintaining a 99% survival rate in STGG. The GAMICBANK bead system effectively preserves both robust and sensitive microorganisms at –80 °C. STGG provides superior cryoprotection compared to skim milk, particularly for sensitive bacteria and yeast. In conclusion, the GAMICBANK system serves as a viable alternative method for the preservation of bacterial and yeast isolates.
Keywords
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