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Abstrak :
Latar Belakang: Candida albicans merupakan flora komensal yang dapat berubah menjadi virulen pada keadaan tertentu yang dipengaruhi oleh faktor predisposisi dan virulensi. Salah satu faktor virulensi C. albicans  adalah kemampuan membentuk biofilm dengan gambaran morfologi yang berubah pada setiap fasenya. Pembentukan biofilm dapat meningkatkan resistensi terhadap agen antijamur. Temulawak merupakan tanaman obat unggulan Indonesia yang diketahui memiliki khasiat antijamur. Tujuan: Mengetahui perkembangan berbagai fase biofilm C. albicans ATCC 10231 setelah paparan ekstrak etanol temulawak (Curcuma xanthorrhiza Roxb.) Metode: Uji MTT-assay digunakan untuk mengetahui konsentrasi minimum ekstrak etanol temulawak dalam menghambat pembentukan biofilm C. albicans (KHBM₅₀). Gambaran mikroskopis perkembangan biofilm C. albicans diobservasi dengan menggunakan  Scanning Electron Microscope (SEM). Hasil: Nilai Konsentrasi Inhibisi Biofim Minimal (KHBM₅₀) ekstrak etanol temulawak terhadap biofilm C. albicans ATCC 10231 pada fase awal (adhesi dan proliferasi), fase menengah, dan fase maturasi berturut turut adalah 25%, 35%, dan 40%. Kemampuan ekstrak etanol temulawak dalam menghambat perkembangan biofilm C. albicans  menurun seiring dengan peningkatan fase biofilm.  Pada fase adhesi, morfologi C. albicans ATCC 10231 yang dipaparkan ekstrak etanol temulawak dan nystatin masih berbentuk blastospora, berbeda dengan kontrol negatif yang sudah menunjukkan germinasi. Pada fase proliferasi, menengah, dan maturasi C. albicans ATCC 10231 yang dipaparkan temulawak maupun nystatin menunjukkan adanya pertumbuhan hifa yang lebih pendek namun dengan jumlah dan densitas yang jauh lebih sedikit jika dibanding dengan kontrol negatif. Kesimpulan: Ekstrak etanol temulawak mempengaruhi viabilitas C. albicans ATCC 10231 dan menghambat perkembangan biofilm C. albicans ATCC 10231 dengan cara menghambat pertumbuhan hypha serta menurunkan densitas biofilm. Semakin meningkat fase perkembangan biofilm, dibutuhkan konsentrasi ekstrak etanol temulawak yang lebih tinggi. ...... Background: Candida albicans is a commensal flora that can turn into virulent in certain circumstances that are influenced by predisposing and virulence factors. One of the virulence factors of C. albicans is the ability to form biofilm with morphologic changes in every phase. Biofilm formation can increase resistance towards antifungal agents. Javanese turmeric is an Indonesian medical plant that is reported to have antifungal effect which can inhibit the development of C. albicans biofilm. Objective: To observe the development of Candida albicans ATCC 10231 biofilm formation after exposed to Javanese turmeric ethanol extract (Curcuma xanthorrhiza Roxb.) Method: MTT-assay was used to measure the minimum inhibitory concentration of Javanese turmeric ethanol extract in inhibiting C. albicans ATCC 10231 biofilm formation (MBIC₅₀). The morphological changes of the various stages of C. albicans biofilm were observed using Scanning Electron Microscope (SEM). Results: The Minimum Biofilm Inhibitory Concentration (MBIC₅₀) of Javanese turmeric ethanol extract towards formation of C. albicans biofilm ATCC 10231 in the early phase (adhesion and proliferation), intermediate phase, and maturation phase as follows; were 25%,  35%, and 40% respectively. In the adhesion phase, the morphology of C. albicans ATCC 10231 exposed javanese turmeric ethanol extract and nystatin is still in the form of blastospores, unlike negative controls that have shown germination. In the proliferation, intermediate, and maturation phase C. albicans ATCC 10231 exposed to Javanese turmeric ethanol extract and nystatin showed the growth of shorther hyphae and slightly lesser amounts and densities compared negative controls. The ability of javanese turmeric ethanol extract in inhibiting the development of C. albicans biofilm decreased along with the increased of biofilm phase. Conclusion:  Javanese turmeric ethanol extract affected the viability of C. albicans cells and inhibit the development of C. albicans biofilm by inhibiting the hyphal formation and decreasing the biofilm density.

Abstrak :
Latar Belakang: Temulawak (Curcuma xanthorrhiza Roxb.) merupakan salah satu tanaman obat unggul Indonesia yang memiliki potensi untuk menghambat pembentukan biofilm C. albicans. Faktor virulensi yang dapat menyebabkan C. albicans menjadi fungi patogen diantaranya adalah pembentukan biofilm dan sekresi enzim hidrolitik. Fosfolipase merupakan salah satu enzim hidrolitik yang dapat merusak membran sel inang. Tujuan: Menganalisis aktivitas fosfolipase pada biofilm C. albicans ATCC 10231 fase awal, menengah, dan maturasi yang terhambat ekstrak etanol temulawak (Curcuma xanthorrhiza Roxb.). Metode: Nilai Kadar Hambat Biofilm Minimal (KHBM50) C. albicans ditentukan dengan uji MTT-assay. Ekstrak etanol temulawak dengan konsentrasi sesuai KHBM50 dipaparkan pada biofilm fase awal, menengah, dan maturase. Kontrol negative tidak dipaparkan apapun, kontrol positif dipaparkan Nystatin 100.000 IU. Aktivitas fosfolipase biofilm C. albicans dianalisis dengan mengukur proporsi antara diameterzona presipitasi dengan diameter koloni C. albicans pada medium Egg Yolk Agar (EYA). Hasil: Nilai KHBM50 ekstrak etanol temulawak terhadap biofilm C. albicans ATCC 10231 pada fase awal, fase menengah, dan fase maturasi berturut-turut adalah 25%, 30%, dan 35%. Pada kontrol positif, aktivitas fosfolipase biofilm C. albicans fase awal, fase menengah, dan fase maturasi bernilai 1. Aktivitas fosfolipase biofilm C. albicansfase awal, fase menengah, dan fase maturasi yang terhambat ekstrak etanol temulawak berturut-turut 0.84, 0.80, dan 0.83. Pada kontrol negatif, aktivitas enzim fosfolipase biofilm C. albicans fase awal, fase menengah, dan fase maturasi berturut-turut 0.59, 0.57, dan 0.57. Kesimpulan: Terdapat kecenderungan penurunan aktivitas enzim fosfolipase pada biofilm C. albicans yang terhambat > 50% ekstrak etanol temulawak.

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Background: Javanese turmeric (Curcuma xanthorrhiza Roxb.) is one of medical plant from Indonesia that has potency to inhibit biofilm formation of C. albicans. Biofilm formation and hydrolyticenzymes are two among manyvirulence factors of C. albicans. Phospholipaseisone of hydrolyticenzymesthat could degrade the hostcell membrane. Objective: To observe the activities ofphospholipase in early phase, intermediate phase, and maturation phase of biofilm C. albicans ATCC 10231 that has been inhibited by Javanese turmeric ethanolic extract. Method: MTT-assay wasused to measure the minimum biofilm inhibitory concentration (MBIC50) of C. albicans ATCC 10231in three phases of C. albicans biofilm. Those concentrations were used to observe phospholipase activities of biofilm in the relevant phases. The negative control were not exposed to anything, while the positive control were exposed to Nystatin 100.000 IU. Phospholipase activities were determined bymeasuring the proportion of precipitation zone diameter and C. albicans colony diameter onan egg yolk-agar medium. Results: The MBIC50of Javanese turmeric ethanolic extract towards formation of C. albicans biofilm ATCC 10231 in early phase, intermediate phase, and maturation phase were 25%, 30%, and 35%, respectively. Phospholipase activities value in early phase, intermediate phase, and maturation phase of C. albicans biofilm exposed by Nystatin were 1. Phospholipase activities value in early phase, intermediate phase, and maturation phase of C. albicans biofilms exposed by Javanese turmeric ethanolic extract were 0.84, 0.80, and 0.83, respectively. Phospholipase activities value in early phase, intermediate phase, and maturation phase of unexposed C. albicans biofilm were 0.59, 0.57, and 0.57, respectively. Conclusion: There istendency of decreased phospholipase activity in early phase, intermediate phase, and maturation phase of biofilm C. albicans that has been inhibited by Javanese turmericethanolic extract.

Abstrak :
Pendahuluan: Salah satu faktor virulensi utama yang mempengaruhi perubahan Candida albicans(C. albicans) dari flora komensal menjadi patogen adalah pembentukan biofilm. Pada biofilm fase maturasi, C. albicansmenjadi lebih resisten terhadap agen antifungal. Telah dibuktikan efek inhibisi ekstrak etanol temulawak (Curcuma xanthorrhiza Roxb.) terhadap pertumbuhan biofilm C. albicans. Tujuan: Menganalisis gambaran TEM sel C. albicans ATCC 10231pada biofilm fase maturasi yang terinhibisi ekstrak etanol temulawak (EET). Metode: Penelitian ini dibagi dua kelompok yaitu kelompok perlakuan dan tanpa perlakuan. Sebanyak 100μL suspensi C. albicans ATCC 10231dalam 96-well-plate diinkubasi selama 1.5 jam pada 370C,kemudian diaspirasi dan dibilas menggunakan PBS. Pada kelompok perlakuan dipapar 100μLEET dengan konsentrasi KHBM50(35%), sedangkan kelompok tanpa perlakuan tidak dipapar EET. Kedua kelompok di inkubasi kembali hingga 48 jam, kemudian dipindahkan ke Eppendorf tube untuk difiksasi dalam 2.5% glutaraldehyde dan dilakukan pemeriksaan TEM. Kontrol positif dipapar nystatin oral suspension. Hasil: Pemeriksaan TEM pada kelompok perlakuan, sel C. albicansATCC 10231 pada biofilm fase maturasi yang terpapar ekstrak etanol temulawak terlihat perubahan gambaran ultrastruktur yang berupa perubahan bentuk sel, penebalan dinding sel, pembesaran vakuola, dan iregularitas sitoplasma berikut organel-organel di dalamnya seperti disorganisasi pada nukleus, mitokondria, dan retikulum endoplasma. Sedangkan pada kelompok tanpa perlakuan menunjukan gambaran sel C. albicans ATCC 10231 normal. Kesimpulan: Pemeriksaan TEM dapat menunjukkan perubahan sel C. albicans ATCC 10231 pada biofilm fase maturasi yang terinhibisi ekstrak etanol temulawak. ...... Introduction: One of main virulence factor that influence the alteration of Candida albicans (C. albicans) from commensal flora into pathogenic flora is biofilm formation. At the maturation phase, C. albicans ismore resistant to antifungal agent. It has been proven that there is an inhibition effect of Javanese turmeric (Curcuma xanthorrhiza Roxb) on the growth of C. albicans biofilm. Objective: To analyze the TEM image of C. albicans cell on the maturation phase of biofilm inhibit by Javanese turmeric ethanol extract. Method: This research divided into two groups, there were treated group and untreated group. A 100μLC. albicans ATCC 10231 suspension on a 96-well-plate incubated for 1.5 hour at 370C, and then aspirated and washed by phosphate buffer saline (PBS). The treated group was exposed to 100μL Javanese turmeric ethanol extract as MBIC50 concentration (35%), while the untreated group was not exposed to Javanese turmeric ethanol extract. Both groups were incubated until 48 h, and then moved into the Eppendorf tube and fixed with glutaraldehyde 2.5% to examine using TEM. The positive control was exposed to nystatin oral suspension. Result: Compared to the negative control, C. albicans cell on biofilm maturation phase treated by Javanese turmeric extract ethanol extract shows the alteration on the its ultra structure. The alteration showed in shape, the thickness of cell wall, the enlargement of vacuole, and irregularity of cytoplasm include the organelles in it such as disorganization on nucleus, cytoplasm, and endoplasmic reticulum. Conclusion: TEM examination showed the alteration of C. albicans ATCC 10231 cell on maturation phase biofilm inhibited by Javanese turmeric ethanol extract.

Abstrak :
Latar Belakang: Temulawak (Curcuma xanthorrhiza Roxb.) merupakan tanaman obat asli Indonesia yang mengandung zat antijamur. Faktor virulensi berperan penting dalam proses infeksi Candida albicans, salah satunya adalah sekresi enzim fosfolipase yang dapat merusak membran sel inang. Tujuan: Penelitian ini bertujuan untuk menganalisis pengaruh penghambatan dan pemberantasan aktivitas enzim fosfolipase pada fase awal, intermediet, dan pematangan biofilm C. albicans ATCC 10231. Metode: Efek penghambatan ekstrak etanol temulawak diamati dengan menginkubasi C. albicans selama 1,5 jam, kemudian dipapar ekstrak KHBM50. etanol temulawak kemudian diinkubasi selama 6, 24, dan 48 jam untuk mencapai fase awal, intermediet, dan pematangan biofilm C. albicans. Efek eradikasi diamati dengan menginkubasi C. albicans selama 6, 24, dan 48 jam, kemudian dipapar KEBM50 EET dan diinkubasi pada suhu 37ºC selama 24 jam. KHBM50 dan KEBM50 EET (kelompok perlakuan), nistatin 100.000 IU (kontrol positif), dan SDB (kontrol negatif). Aktivitas enzim fosfolipase dianalisis berdasarkan luas zona pengendapan yang terbentuk pada agar kuning telur. Hasil: KHBM50 EET pada fase awal 15%, intermediate 15%, dan pematangan 25%. Nilai KEBM50 EET untuk ketiga fase biofilm C. albicans adalah 35%. Pada kontrol positif baik inhibisi maupun eradikasi, tidak terlihat adanya zona presipitasi pada ketiga fase biofilm C. albicans. Sementara itu, kelompok penghambatan dan pemberantasan menunjukkan ukuran zona pengendapan yang lebih kecil jika dibandingkan dengan kelompok kontrol negatif pada ketiga fase biofilm C. albicans. Kesimpulan: Aktivitas enzim fosfolipase cenderung menurun pada fase awal, menengah, dan pematangan biofilm C. albicans setelah penghambatan dan eradikasi ekstrak etanol temulawak.
Background: Temulawak (Curcuma xanthorrhiza Roxb.) is a medicinal plant native to Indonesia that contains antifungal substances. Virulence factors play an important role in the process of Candida albicans infection, one of which is the secretion of phospholipase enzymes that can damage host cell membranes. Objective: This study aimed to analyze the effect of inhibition and eradication of phospholipase enzyme activity in the early, intermediate, and maturation phases of C. albicans ATCC 10231 biofilm. Methods: The inhibitory effect of temulawak ethanol extract was observed by incubating C. albicans for 1.5 hours, then exposed to KHBM50 extract. Temulawak ethanol was then incubated for 6, 24, and 48 hours to reach the initial, intermediate, and maturation phases of the C. albicans biofilm. The eradication effect was observed by incubating C. albicans for 6, 24, and 48 hours, then exposed to KEBM50 EET and incubated at 37ºC for 24 hours. KHBM50 and KEBM50 EET (treatment group), nystatin 100,000 IU (positive control), and SDB (negative control). The activity of the phospholipase enzyme was analyzed based on the area of ​​the deposition zone formed on egg yolk agar. Results: KHBM50 EET in early phase 15%, intermediate 15%, and maturation 25%. The KEBM50 EET value for the three phases of the C. albicans biofilm was 35%. In the positive control, both inhibition and eradication, no precipitation zones were seen in the three phases of the C. albicans biofilm. Meanwhile, the inhibition and eradication groups showed a smaller deposition zone size when compared to the negative control group in all three phases of the C. albicans biofilm. Conclusion: The activity of the phospholipase enzyme tends to decrease in the early, intermediate, and maturation phases of C. albicans biofilm after inhibition and eradication of ethanol extract of temulawak.

Abstrak :
Penyakit infeksi fungi merupakan penyakit yang sering ditemukan di Indonesia. Pencarian antibiotik yang tepat untuk penyakit infeksi fungi masih terus dilakukan. Pseudomonas azotoformans UICC B-91 berpotensi sebagai antimikroba. Tujuan penelitian untuk mengetahui mekanisme penghambatan metabolit P. azotoformans UICC B-91 dari medium terhadap fungi patogen secara morfologi. Hasil penelitian menunjukkan senyawa metabolit P. azotoformans UICC B-91 mampu menghambat C. albicans ATCC 10231 dan T. mentagrophytes. Zona inhibisi larutan medium P. azotoformans UICC B-91 terhadap C. albicans ATCC 10231 konsentrasi 100 mg/mL dan 80 mg/mL memiliki daya hambat kuat, konsentrasi 60 mg/mL memiliki daya hambat sedang. Zona inhibisi terhadap T. mentagrophytes konsentrasi 100 mg/mL dan 80 mg/mL memiliki daya hambat kuat, konsentrasi 60 mg/mL memiliki daya hambat sedang. Pengamatan mikroskopis C. albicans ATCC 10231 setelah penambahan medium P. azotoformans UICC B-91 di bawah Scanning Electron Microscope (SEM) mengalami perubahan permukaan sel yeast menjadi tidak rata. Pengamatan mikroskopis T. mentagrophytes setelah penambahan medium P. azotoformans UICC B-91 mengalami konstriksi mikrokonidia. Mekanisme penghambatan metabolit dari medium P. azotoformans UICC B-91 terhadap C. albicans ATCC 10231 diduga melalui difusi yang mengganggu fungsi membran sel atau menghambat transisi bentuk yeast ke bentuk hifa, sedangkan terhadap T. mentagrophytes diduga mengganggu dinding sel dan mengganggu fungsi membran sel. ......Fungal infection is a disease that is often found in Indonesia. The search for appropriate antibiotics for fungal infections is still ongoing. Pseudomonas azotoformans UICC B-91 has potential as an antimicrobial. The purpose of this study was to determine the mechanism of inhibition of the metabolite of P. azotoformans UICC B-91 from the medium against morphologically pathogenic fungi. The results showed that the metabolite compound P. azotoformans UICC B-91 was able to inhibit C. albicans ATCC 10231 and T. mentagrophytes. Inhibition zone of medium solution of P. azotoformans UICC B-91 against C. albicans ATCC 10231 with a concentration of 100 mg/mL and 80 mg/mL had a strong inhibitory effect, a concentration of 60 mg/mL had a moderate inhibitory effect. The zone of inhibition for T. mentagrophytes at concentrations of 100 mg/mL and 80 mg/mL had a strong inhibitory effect, a concentration of 60 mg/mL had moderate inhibition. Microscopic observation of C. albicans ATCC 10231 after the addition of P. azotoformans UICC B-91 medium under a Scanning Electron Microscope (SEM) changed the yeast cell surface to become uneven. Microscopic observation of T. mentagrophytes after the addition of P. azotoformans UICC B-91 medium experienced constriction of microconidia. The mechanism of inhibition of metabolites from P. azotoformans UICC B-91 medium against C. albicans ATCC 10231 is thought to be through diffusion which disrupts cell membrane function or inhibits the transition from yeast to hyphal form, while against T. mentagrophytes it is thought to disrupt cell walls and disrupt cell membrane function.

Abstrak :
Pendahuluan: Perubahan sistem pertahanan pejamu atau kondisi rongga mulut dapat menyebabkan infeksi Candida albicans (C. albicans) yang disebut kandidiasis oral. Xanthorrhizol adalah komponen aktif Temulawak (Curcuma xanthorrhiza Roxb.) yang memiliki efek antijamur. Gambar Scanning Electron Microscope (SEM) sel C. albicans yang terpapar xanthorrhizol menunjukkan penonjolan sitoplasma dan adhesi antar sel. Tidak diketahui bagaimana perubahan ultrastruktural pada sel C. albicans. Tujuan: Menggunakan TEM untuk menganalisis perubahan ultrastruktur sel C. albicans pada biofilm fase pematangan pasca-inhibisi dengan Ekstrak Etanol Temulawak (EET). Metode: Biofilm C. albicans ATCC 10231 yang ditumbuhkan pada kawat selama 1,5 jam dihambat dengan 25% EET selama 48 jam. Kelompok kontrol positif tidak diberi nistatin sedangkan kelompok kontrol negatif tidak diberi apa-apa. Sampel difiksasi dengan 2,5% glutaraldehid, didehidrasi dengan etanol, dan dibenamkan dalam resin spurr sebelum diamati dengan TEM. Hasil: Terlihat distorsi dinding sel, membran plasma, dan organel serta invaginasi membran plasma. Kesimpulan: Penghambatan biofilm C. albicans ATCC 10231 oleh 25% EET menyebabkan kerusakan pada ultrastruktur sel C. albicans ATCC 10231.

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Changes of hosts defense system or oral condition causing infection of Candida albicans (C. albicans) is called oral candidiasis. Xanthorrhizol is an active component of javanese turmeric (Curcuma xanthorrhiza Roxb.) which has antifungal effect. C. albicans cells Scanning Electron Microscope (SEM) image exposed to xanthorrhizol showed cytoplasm protrusion and clumping. The ultrastructure changes inside the C. albicans cell is not known yet. Objective: Using TEM to analyse the ultrastructure image of C. albicans ATCC 10231 cells in maturation phase of biofilm inhibited by 25% EET. Methods: C. albicans ATCC 10231 biofilm which had been cultured on wire for 1.5 hours was inhibited by 25% EET. Group of positive control was exposed by nystatin whereas group of negative control was exposed to nothing. Sampel was being fixated with glutaraldehyde 2.5%, dehydrathed by ethanol, and embedded inside spurrs resin before being observed with TEM. Results: Cell wall, plasma membrane, and organelles distortion, along with plasma membrane invagination. Conclusion: C. albicans ATCC 10231 biofilm inhibition with 25% EET caused damages in C. albicans ATCC 10231 cells ultrastructure.