Perubahan Kandungan Asam Lemak Rantai Pendek dan Bakteri Enterobacteriaceae Pada Feses Remaja Usia 15-19 Tahun yang Mengonsumsi Probiotik Multistrain

Mariyatun, Prof. Dr. Ir. Donny Widianto; Dr. Dian Anggraini Suroto, STP, MP, M.Eng.

2025 | Tesis | S2 Bioteknologi

Probiotik multistrain Lactiplantibacillus plantarum subs. plantarum (strain Dad-13 dan FNCC-0250) dan Lacticaseibacillus paracasei GMRMP-001 terbukti mampu bersinergi, memiliki sifat antibakteri dan menghasilkan asam lemak rantai pendek (ALRP) pada penelitian menggunakan hewan coba. Tujuan penelitian ini adalah untuk mengetahui pengaruh pemberian probiotik multistrain L. plantarum (strain Dad-13 dan FNCC-0250) dan L. paracasei GMRMP-001 terhadap ALRP dan Enterobacteriaceae feses remaja sehat. Metode yang digunakan adalah double-blind placebo-controlled randomized controlled trial (RCT) yang melibatkan 60 partisipan remaja sehat usia 17-25 tahun. Partisipan diacak menjadi 2 kelompok yaitu kelompok probiotik dan kelompok placebo. Penelitian dilakukan selama 30 hari, dimana kelompok probiotik mengonsumsi probiotik multistrain 10¹⁰ CFU dalam 0,25 g susu skim, kelompok placebo mengonsumsi 0,25 g susu skim. Partisipan melakukan pencatatan konsumsi produk penelitian dan pola makan selama penelitian. Pada hari ke-1 ± 1 dan hari ke-30 ± 1 partisipan diminta untuk mengumpulkan sampel feses. Jumlah koloni bakteri dihitung menggunakan metode total plate count (TPC), bakteri asam laktat dihitung dengan menumbuhkan pada media De Man Ragosa and Sharpe Agar (MRSA) dan Lactitol LBS vancomycin (LLV). Jumlah L. plantarum dihitung menggunakan media Lactobacillus plantarum selective medium (LPSM) dan violet red bile Agar (VRBGA) untuk Enterobacteriaceae. Jumlah L. plantarum dan L. paracasei juga dianalisa menggunakan metode quantitative polymerase chain reaction (qPCR). Konsentrasi ALRP dianalisa menggunakan Gas Chromatography (GC). Hasil penelitian menunjukkan bahwa jumlah koloni hidup kelompok bakteri asam laktat yang ditumbuhkan pada media LLV meningkat signifikan setelah mengonsumsi probiotik. Hal ini juga diikuti dengan peningkatan signifikan L. plantarum, dimana peningkatan keduanya berbeda nyata dibandingkan plasebo. Lebih lanjut, Enterobacteriaceae mengalami penurunan signifikan pada kelompok probiotik. Dengan metode PCR, jumlah L. plantarum dan L. paracasei mengalami peningkatan secara signifikan dan berbeda nyata dibandingkan kelompok plasebo. Konsumsi probiotik multistrain juga meningkatkan konsentrasi asam asetat secara signifikan dibandingkan kelompok plasebo. Hasil penelitian menunjukkan bahwa konsumsi probiotik multistrain L. plantarum subs. plantarum (strain Dad-13 dan FNCC-0250) dan L. paracasei GMRMP-001 selama 30 hari mampu menurunkan Enterobacteriaceae dan meningkatkan asam asetat.

The multistrain probiotics Lactiplantibacillus plantarum subsp. plantarum (strains Dad-13 and FNCC-0250) and Lacticaseibacillus paracasei GMRMP-001 have demonstrated synergistic effects, exhibit antibacterial properties, and the ability to produce short-chain fatty acids (SCFAs) in animal study. The aim of this study was to determine the effects of administering the multistrain probiotic, consist of L. plantarum (strains Dad-13 and FNCC-0250) and L. paracasei GMRMP-001 on the fecal Enterobacteriaceae and SCFA concentrations in healthy adolescents. A double-blind placebo-controlled randomized controlled trial (RCT) was conducted involving 60 healthy adolescent participants aged 17-25 years. Participants were randomly assigned to either a probiotic or placebo group. Over a 30-day intervention period, the probiotic group consumed multistrain probiotics (10¹? CFU) suspended in 0.25 g of skim milk, while the placebo group consumed 0.25 g of skim milk without the addition of probiotics strain. Participants recorded their daily product intake and dietary pattern throughout the study period. Fecal samples were collected at day 1 ± 1 and day 30 ± 1. Total bacterial colony counts were assessed using the total plate count (TPC) method. Lactic acid bacteria (LAB) were cultured on de Man, Rogosa and Sharpe Agar (MRSA) and Lactitol LBS Vancomycin (LLV) media. L. plantarum was enumerated using L. plantarum selective medium (LPSM), and Enterobacteriaceae were quantified using violet red bile agar (VRBGA). Quantitative polymerase chain reaction (qPCR) was also used to determine the levels of L. plantarum and L. paracasei. SCFA concentrations were analyzed using gas chromatography (GC). The results showed a significant increase in the viable LAB group, particularly those grown on LLV media, following probiotic consumption. This was accompanied by a significant increase in L. plantarum, with both showing statistically significant differences compared to the placebo group. Furthermore, fecal Enterobacteriaceae showed a significant decrease in the probiotic group. qPCR analysis confirmed a significant increase in both L. plantarum and L. paracasei compared to the placebo. In addition, acetic acid concentrations were significantly higher in the probiotic group. In conclusion, 30 days of supplementation with a multistrain probiotic L. plantarum subs. plantarum (strain Dad-13 and FNCC-0250) and L. paracasei GMRMP-001 significantly reduced fecal Enterobacteriaceae and increased acetic acid concentration in healthy adolescents.

Kata Kunci : Probiotik Multistrain, Asam Lemak Rantai Pendek, Enterobacteriaceae, Lactobacillus plantarum, Lactobacillus paracasei