"The Effect of Temperature for Producing Thayer-Martin Agar Media Using" by Ika Ningsih and Kayla Adyapratista Sumadiyo
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Indonesian Journal of Medical Chemistry and Bioinformatics

Abstract

Neisseria gonorrhoeae is a Gram-negative bacterium in the form of diplococci that infects mucous membranes through direct inoculation, is fastidious, and requires special media for growth because it is susceptible to heating and dryness, so it cannot survive long outside its host. Thayer-Martin agar media is a selective culture medium for the diagnosis of Neisseria gonorrhoeae bacterial infections enriched with sheep blood. The purpose of this study was to determine the optimum temperature for various temperature variations of making Thayer-Martin agar when mixing with sheep blood, compared to the temperature according to the manufacturer's procedure. Determining this ideal temperature may help improve the accuracy of the diagnosis of Neisseria gonorrhoeae infection. This study used an experimental laboratory design. Sheep blood was mixed into Thayer-Martin agar at several different temperatures: 56°C, 65°C, 75°C, and 85°C. After agar preparation, Neisseria gonorrhoeae ATCC 43069 was inoculated onto the agar with McFarland standards of 0.5, 1.0, 2.0, and 3.0. The plates were then incubated under appropriate conditions to observe bacterial growth. Neisseria gonorrhoeae showed varying growth patterns at different Thayer-Martin agar preparation temperatures, which was confirmed using ANOVA analysis with a p-value of 0.002. At 56°C, full growth was observed consistently across all McFarland turbidity standards (0.5, 1.0, 2.0, and 3.0). At 65°C, growth varied with a combination of full and half-full growth in all standards. However, temperatures of 75°C and 85°C resulted in significantly reduced growth, showing only half-full growth in all standards. Post hoc tests revealed that agar preparation at 56°C provided the most favorable conditions for Neisseria gonorrhoeae growth compared to the other temperatures tested. This study found that 56°C provided optimal conditions for the growth of Neisseria gonorrhoeae because it resulted in maximum bacterial growth. Higher temperatures, such as 65°C, 75°C, and 85°C, resulted in reduced bacterial growth, possibly due to protein denaturation or bacterial cell damage at these temperatures.

Bahasa Abstract

Neisseria gonorrhoeae adalah bakteri Gram-Negative berbentuk diplokokus yang menginfeksi selaput lendir melalui inokulasi langsung, sulit untuk dikultur, dan memerlukan media khusus untuk pertumbuhannya, seperti media agar Thayer-Martin, yang merupakan media kultur selektif untuk diagnosis infeksi bakteri Neisseria gonorrhoeae yang diperkaya dengan darah domba. Variasi suhu digunakan untuk mencampur media Thayer-Martin dengan darah domba \, sehingga standar suhu optimum untuk pertumbuhan Neisseria gonorrhoeae dapat ditemukan. Tujuan dari penelitian ini adalah untuk menentukan suhu optimum untuk berbagai variasi suhu dalam pembuatan media agar Thayer-Martin yang dicampur dengan darah domba, dibandingkan dengan menggunakan suhu sesuai prosedur pabrikan, sehingga diharapkan suhu ideal ini dapat membantu meningkatkan akurasi diagnosis Neisseria gonorrhoeae. Studi ini menggunakan desain laboratorium eksperimental. Darah domba dicampurkan ke dalam agar Thayer-Martin menggunakan beberapa suhu yang berbeda, yaitu 56°C, 65°C, 75°C, dan 85°C. Kemudian Neisseria gonorrhoeae ATCC 43069 diinokulasi ke dalam media agar menggunakan standar kekeruhan McFarland 0,5, 1,0, 2,0, dan 3,0. Kemudian, diinokulasi dalam inkubator untuk melihat pertumbuhan bakteri. Bakteri Neisseria gonorrhoeae menunjukkan pola pertumbuhan yang bervariasi pada suhu yang berbeda. Pada suhu 56°C, pertumbuhan koloni bakteri meningkat pesat pada semua standar kekeruhan yang diuji (McFarland 0.5, 1.0, 2.0, dan 3.0). Namun, pada suhu 65 °C, pertumbuhan koloni Neisseria gonorrhoeae menunjukkan variabilitas; beberapa standar kekeruhan menunjukkan pertumbuhan koloni yang signifikan, sementara yang lain menunjukkan pertumbuhan yang terbatas atau tidak ada pertumbuhan sama sekali. Suhu 75°C dan 85°C menghasilkan pertumbuhan koloni bakteri yang tidak terlalu banyak pada semua standar kekeruhan McFarland. Uji post hoc menunjukkan bahwa pembuatan media agar pada suhu 56°C memberikan kondisi yang paling menguntungkan untuk pertumbuhan Neisseria gonorrhoeae dibandingkan dengan suhu lainnya yang diuji. Studi ini menemukan bahwa 56°C menyediakan kondisi optimal untuk pertumbuhan Neisseria gonorrhoeae karena menghasilkan pertumbuhan bakteri maksimum. Suhu yang lebih tinggi, seperti 65°C, 75°C, dan 85°C, menghasilkan pertumbuhan bakteri yang lebih sedikit. Hal ini kemungkinan disebabkan oleh denaturasi protein atau kerusakan sel bakteri pada suhu-suhu ini.

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