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Karakteristik Fisikokimia dan Uji Aktivitas Antimikroba Bakteriosin dari Isolat Bakteri Asam Laktat 15B hasil Isolasi Kolon Sapi Bali Lestari, Ni Kadek Lyming; Suardana, I Wayan; Sukrama, I Dewa Made
Buletin Veteriner Udayana Vol. 11 No. 1 Pebruari 2019
Publisher : The Faculty of Veterinary Medicine, Udayana University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (166.044 KB) | DOI: 10.24843/bulvet.2019.v11.i01.p11

Abstract

Lactic acid bacteria (LAB) is a group of Gram-positive bacteria, in shape of coccus or bacilli and catalase-negative and also have the ability to produce lactic acid. LAB is also a microorganism which is not toxic and has the ability to produce antimicrobial compounds, known as bacteriocins. This research aimed to find out the physicochemical characteristic and the antimicrobial activities of bacteriocin isolated from isolates 15B originated from the bali cattle’colon. This study began by reculturing of isolates 15B and followed by cultivating the isolates, isolating, producing, and precipitating of bacteriocins. The physical and chemical properties of the bacteriocins were also tested before, antimicrobial activity test was conducted. The result of the cultivation showed that isolates 15B is a lactic acid bacteria characterized by the result of Gram staining as a Gram positive, catalase negative and grown on anaerobic condition. The result of chemical test, showed that bacteriocins producing isolates 15B were positive on Ninhydrin test, negative on Molisch test, and positive on Lowry test with 0,11 µg/ml concentration. Physical properties test using SDS PAGE showed negative result and antimicrobial activity test showed that the bacteriocin from isolates 15B has 35,82% effectiveness of inhibition.
Identifikasi Bakteri Asam Laktat Isolat 9A dari Kolon Sapi Bali sebagai Probiotik melalui Analisis Gen 16S rRNA Widyadnyana, Dewa Gede Agung; Sukrama, I Dewa Made; Suardana, I Wayan
Jurnal Sain Veteriner Vol 33, No 2 (2015): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (532.142 KB) | DOI: 10.22146/jsv.17923

Abstract

Bakteri asam laktat (BAL) merupakan bakteri yang mampu memfermentasi laktosa dan menghasilkan asam laktat sebagai produk utamanya. BAL berpotensi dalam memproduksi bakteriosin dan bersifat probiotik. Disisi lain, sapi bali diketahui memiliki banyak keunggulan salah satunya mampu memanfaatkan hijauan yang kurang bergizi, dan mempunyai daya cerna yang baik terhadap pakan, sehingga diharapkan dapat ditemukan BAL yang memiliki keunggulan spesifik. Penelitian ini bertujuan untuk dapat mengetahui aktivitas antimikroba BAL isolat9A asal kolon sapi bali sehingga bisa di manfaatkan sebagai kandidat probiotik, serta untuk menganalisis susunan nukleotida dan gambaran pohon filogenetiknya. Penelitian diawali dengan kultivasi isolat pada media MRS broth, dilanjutkan dengan uji katalase dan pewarnaan Gram. Isolat BAL yang sudah terkonfirmasi diuji aktivitas antimikrobanya dengan bakteri patogen (Escherichia coli dan Staphylococcus aureus). Analisis molekuler dilakukan dengan amplifikasi gen 16S rRNA menggunakan primer B27F dan U1492R. Hasil penelitian menunjukkan aktivitas antimikroba bakteri asam laktat isolat 9A mampu menghambat pertumbuhan S. aureus antara 37,56 - 61,47%, dengan rata-rata 50.35% jika dibandingkan dengan kontrol antibiotika. Dalam penelitian ini BAL tidak menunjukkan terjadinya penghambatan pada pertumbuhan E. Coli. BAL isolat 9A teridentifikasi sebagai Streptococcus bovis dengan tingkat similaritas 99% dan berada satu cluster denganspesies Streptococcus bovis dengan nilai bootstrap sebesar 100.
Optimasi Duplex PCR untuk Deteksi Simultan Gen Penyandi Faktor Virulensi ompW dan ctxA Vibrio cholerae Praja, Rian Ka; Sukrama, I Dewa Made; Fatmawati, Ni Nengah Dwi; Hidayati, Wahyu
Indonesia Medicus Veterinus Vol 5 (5) 2016
Publisher : Faculty of Veterinary Medicine, Udayana University

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Abstract

Vibrio cholerae merupakan salah satu agen foodborne disease yang dapat ditularkan melalui seafood. Penelitian ini bertujuan untuk optimasi gen penyandi faktor virulensi outer membrane protein W (ompW) dan cholerae toxin subunit A (ctxA) menggunakan teknik Duplex Polymerase Chain Reaction (dPCR). Dua bakteri V. cholerae O1 serotipe Ogawa dan Inaba digunakan pada penelitian ini. Proses isolasi DNA dilakukan menggunakan metode Boil Cell Extraction (BCE). dPCR dilakukan menggunakan dua pasang primer (forward dan reverse) ompW-F, ompW-R dan ctxA-F, ctxA-R dengan panjang produk masing-masing 588 bp dan 302 bp. Tahap optimasi yang dilakukan dalam proses dPCR ini meliputi variasi suhu annealing, variasi konsentrasi primer serta variasi volume DNA template kemudian deteksi produk dPCR dilakukan dengan elektroforesis pada gel agarosa 1,5% dan divisualisasi menggunakan alat Gel DocTM XR (Bio-Rad). Hasil penelitian menunjukkan komposisi reaksi dPCR yang terbaik untuk mendeteksi gen ompW dan ctxA secara simultan terdiri dari PCR mix (Promega) 12,5 ?L, primer ompW-F, ompW-R 0,5 ?M, primer ctxA-F, ctxA-R 0,3 ?M, nuclease free water 6,5 ?L dan DNA template 2 ?L sehingga volume total menjadi 25 ?L. Kondisi mesin PCR terdiri dari pre-denaturasi 95oC selama 2 menit (1 siklus) diikuti oleh denaturasi 95oC selama 1 menit, annealing 53oC selama 1 menit, extension 72oC selama 1 menit (35 siklus), dan post-extension 72oC selama 5 menit (1 siklus). Dapat disimpulkan bahwa dPCR dapat digunakan untuk deteksi simultan gen penyandi faktor virulensi ompW dan ctxA V. cholerae.
POLA KEPEKAAN BAKTERI Salmonella typhi TERISOLASI DARI DARAH TERHADAP SIPROFLOKSASIN DAN SEFTRIAKSON DI RSUP SANGLAH PERIODE JANUARI 2015 - MARET 2017 Yogita, Saraswati P.; Hendrayana, Made Agus; Sukrama, I Dewa Made
E-Jurnal Medika Udayana Vol 7 No 12 (2018): Vol 7 No 12 (2018): E-Jurnal Medika Udayana
Publisher : Universitas Udayana

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Abstract

Salmonella typhi merupakan bakteri gram negatif yang dapat menyebabkan demam tifoid, septicemia (infeksi bakteri di dalam aliran darah) dan gastroenteritis. Penyebaran strain resisten S. typhi terhadap antibiotika lini pertama dan kedua yakni Siprofloksasin dan Seftriakson menjadi perhatian yang sangat serius di seluruh dunia, khususnya di Indonesia. Untuk itu, perlu dilakukan penilitan lebih lanjut untuk mendapatkan data agar penggunaan antibiotika secara rasional, efektif serta untuk mencegah terjadinya multidrug-resistant S. typhi (MDR-ST) di RSUP Sanglah Denpasar. Penelitian ini merupakan deskriptif observasional dengan rancangan cross sectional. Populasi pada penelitian ini adalah pasien dengan isolat S. typhi yang telah di kultur darah dan dilakukan uji kepekaan kuman di RSUP Sanglah Denpasar pada Januari 2015 hinggga Maret 2017. Pemilihan sampel menggunakan total sampling dengan total sampel 30. Hasil yang diperoleh dari penelitian ini yaitu S. typhi memiliki tingkat sensitivitas sebesar 100% terhadap antibiotika Siprofloksasin maupun Seftriakson sehingga dapat digunakan sebagai antibiotika pilihan untuk therapy pada infeksi S. typhi di RSUP Sanglah Denpasar. Kata Kunci: Salmonella typhi, sensitivitas, Siprofloksasin, Seftriakson
Detection of genes encoding ompW and ctxA of Vibrio cholerae isolated from shrimp and shellfish at Kedonganan fish market, Bali-Indonesia Kusuma Praja, Rian Arinta; Sukrama, I Dewa Made; Fatmawati, Ni Nengah Dwi
Oceana Biomedicina Journal Vol 2, No 1 (2019): Oceana Biomedicina Journal
Publisher : Universitas Hang Tuah

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (251.1 KB) | DOI: 10.30649/obj.v2i1.23

Abstract

Contamination of pathogenic bacteria in food can lead to the emergence of foodborne disease. One of foodborne disease which often occurs in some developing countries such as Africa, Southeast Asia, and Latin America is cholera which is caused by Vibrio cholerae. The disease is transmitted through beverages and food, especially contaminated seafood. V. cholerae has several virulence factors including the outer membrane protein W (ompW) and cholerae toxin (ctx).The ompW acts as a protective barrier and can also be used as a marker specific species of V. cholerae and cholerae toxin is an enterotoxin responsible for the incidence of diarrhea in a cholera outbreak produced by pathogenic V. cholerae. This study was an observational study to determine the level of contamination of V. cholerae by detecting the outer membrane protein W (ompW) and cholerae toxin subunit A (ctxA) gene of V. cholerae in shrimp and shellfish sold at Kedonganan fish market. Samples were taken using total sampling technique and obtained 24 samples consisting of 14 shrimp samples and 10 shellfish samples. Samples were examined using culture methods and biochemical tests, and then further tested using Duplex Polymerase Chain Reaction (dPCR) to detect ompW and ctxA gene. The dPCR assay results showed 8 out of 14 (57.1%) samples from shrimp and 1 out of 10 (10%) samples from the shellfish positive carried ompW gene, and found no positive samples carrying the ctxA gene in samples derived from shrimp and shellfish. Chi square test analysis results indicated contamination of V. cholerae in shrimp was higher than shellfish based on ompW gene (p<0.05). It can be concluded that the shrimp and shellfish at Kedonganan fish market are contaminated by V. cholerae. Further research is needed to detect the virulence factors besides ompW and ctxA of V. cholerae in seafood. Keywords: Foodborne disease, Vibrio cholerae, ompW gene, ctxA gene, and Duplex Polymerase Chain Reaction (dPCR).
KUALITAS MIKROBIOLOGIS SAMPEL LAWAR MERAH BABI MENGGUNAKAN METODE TOTAL PLATE COUNT Veronica, Sieny; Hendrayana, Made Agus; Sukrama, I Dewa Made
E-Jurnal Medika Udayana Vol 8 No 9 (2019): Vol 8 No 9 (2019): E-Jurnal Medika Udayana
Publisher : Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (419.567 KB) | DOI: 10.24843/MU.2019.V8.i9.P15

Abstract

Lawar merah babi merupakan makanan tradisional Bali yang mudah terkontaminasioleh Staphylococcus aureus karena pengolahannya yang sederhana dan cenderungmenggunakan tangan kosong. Penelitian cross-sectional deskriptif ini bertujuanuntuk mengetahui kualitas mikrobiologis sampel lawar merah babi denganmenggunakan metode Total Plate Count. Terdapat total 12 sampel lawar merahbabi yang diperoleh dimana sampel tersebut kemudian dibuat menjadi tiga variasipengenceran dan dikultur pada media Mannitol Salt Agar. Koloni Staphylococcusaureus dihitung menggunakan metode Total Plate Count sehingga diperolehjumlah koloni dalam satuan CFU/g untuk setiap sampel lawar merah babi. Hasilpenelitian menunjukkan bahwa seluruh sampel lawar merah babi (100%)terkontaminasi oleh Staphylococcus aureus dengan jumlah koloni melebihi batasmaksimum kontaminasi yang diijinkan oleh BPOM pada makanan, yakni 102 CFU/g. Sepuluh dari 12 sampel (83%) bahkan memiliki jumlah hitung kolonimelebihi 105 CFU/g yang dapat menyebabkan produksi enterotoksin. Dapatdisimpulkan bahwa sampel lawar merah babi yang diperoleh memiliki kualitasmikrobiologis yang kurang baik. Oleh karena itu, pemilihan bahan baku yang baikdan pengolahan yang hiegenis sebaiknya dilakukan oleh penjual dan penyajimakanan untuk meningkatkan kualitas lawar merah babi. Kata kunci: Kualitas mikrobiologis makanan, Lawar merah babi, Staphylococcusaureus, Total Plate Count
EFEK KONSUMSI BIO-YOGHURT DARI LACTOBACILLUS RHAMNOSUS SKG34 TERHADAP LIPID PROFILE PADA MANUSIA Pratiwi, I Desak Putu Kartika; Nocianitri, Komang Ayu; Aryanta, Wayan Redi; Ramona, Yan; Sukrama, I Dewa Made; Sujaya, I Nengah
ARCHIVE OF COMMUNITY HEALTH Vol 2 No 1 (2013): Juni (2013)
Publisher : Program Studi Kesehatan Masyarakat Universitas Udayana

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Abstract

This research was carried out to elucidate the eff ect of consumption bio-yoghurt produced byLactobacillus rhamnosus SKG34 on plasma lipid profi les in human. The study was performed usingpre and post analysis. Eleven healthy volunteers aged between 30-48 yeras old were participatedin this study. Each volunteers consumed 100g of bio-yoghurt containing L.rhamnosus SKG34 for 21 days. Blood samples were taken before the start of experiment and after 21 days of comsumption. Total cholesterol was measured using CHOP-POD method, while triglyceride determinedusing glycerol method, LDL and HD determined using homogenous method. Results of thestudy revealed that consumption of the bio-yoghurt lower values of plasma lipid concentrationsin hypercholesterolemia subjects. Total cholesterol, triglyceride and LDL was reduced within4.91%, 13.84%, and 8.24%, respectively, when compare to its concentrations before treatment.However, the plasma lipid concentrations in normal subjects were almost stable during thetreatment periods. Consumption of bio-yoghurt for 21 days had no eff ect on HDL-cholesterollevel. These results indicate that consumption of bio-yoghurt containing L.rhamnosus SKG34specifi cally lowered the concentrations of serum total cholesterol, LDL, and triglyceride onhypercholesterolemia subjects.
XANTHINE OXYDASE INHIBITION OF KOMBUCHA TEA IN HYPERURICEMIA INDUCED WISTAR RAT: decrease of uric acid, malondialdehyde, and 8-hydroxy-2'-deoxyguanosine Sukrama, I Dewa Made
BALI MEDICAL JOURNAL Vol 4 No 1 (2015)
Publisher : BALI MEDICAL JOURNAL

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (150.23 KB)

Abstract

Background: Hyperuricemia is a condition of high level of uric acid in the body due to distortion of purine nucleoside metabolism through hipoxanthin, xanthin, and guanin of basic purine. Objective: to find a cure of hyperuricemia base on the utilization of kombucha tea. Methods: This is a true experimental study by applying posttest only control group design to determine whether kombucha tea inhibit xanthine oxidase in hyperuricemic induced rat reveales by decrease of uric acid, malondialdehyde (MDA), and 8-hydroxy-2’-deoxyguanosine (8-OHdG). In this study, hyperuricemia rat was achieved by intake of high purine diet. Rats were fed with a mixture of 4 g/kg BW of Gnetum gnemon with 50 mL/kg BW of chicken liver ad libitum for 9 days. Treatments in this research are combination of fermentation time of Kombucha tea and volume of this tea, i.e fermentation time 4, 8, and 12 days and the volume are 1 mL and 4 mL. Therefore, there would be seven groups of treatment including control group. ANOVA was then applied to determine the treatment effect with p < 0.05 was concidered significant. Results: This study indicates that kombucha tea has an ability to inhibit xanthine oxidase in hyperuricemic induced rat and decrease uric acid, MDA, and 8-OHdG. This ability was achieved with combination treatment of 12 days fermentation and 4 mL of kombucha intake. Xanthine oxidase, uric acid, MDA, and 8-OHdG levels by this treatment were obtained significantly lower compare to control group. Conclusion: This study proved that kombucha tea was potent to cure hyperuricemia of wistar rat via inhibition of xanthine oxidase produced.
THE EXISTENCE OF VIBRIO CHOLERAE IN INDONESIA: FROM ENVIRONMENTAL TO CLINICAL ASPECTS (A CONCISE REVIEW) Ka Praja, Rian; Pusporini, Anggita Ratri; Rosalina, Reny; Arta, I Wayan Muda Suta; Sukrama, I Dewa Made; Fatmawati, Ni Nengah Dwi
Jurnal PPI Dunia Vol 4 No 1 (2021)
Publisher : OISAA

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Abstract

Vibrio cholerae is an infectious agent causing cholera disease with a high prevalence in various developing countries. V. cholerae is a pathogen with broad spectrum host that can infect humans and animals, especially aquaculture. The existence of this disease in Indonesia has long been identified in several outbreaks. Various reports in Indonesia have succeeded in finding the existence of V. cholerae in the environment, aquaculture, food and beverage, as well as in clinical cases of V. cholerae infection. The presence of V. cholerae in environment such as water source is commonly related with contamination. However, V. cholerae can be found in aquatic environment as this environment is natural habitat for V. cholerae. Thus, aquaculture is prone to be infected with V. cholerae because the presence of this pathogen is abundant in aquatic environment. Contaminated food and beverage are associated with hygiene and sanitation and human is commonly infected after consuming contaminated food or beverage. This brief review has the main focus to discuss the existence of V. cholerae from environmental to clinical aspects found in Indonesia.
Identifikasi Spesies Streptokokus ?-Hemolisis Hasil Isolasi dari Nasal dan Tonsil Babi dengan Uji Basitrasin Suardana, I Wayan; Dinarini, Ni Made Ayu Aryati; Sukrama, I Dewa Made
Buletin Veteriner Udayana Vol. 13 No. 1 Pebruari 2021
Publisher : The Faculty of Veterinary Medicine, Udayana University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2021.v13.i01.p05

Abstract

Streptococcosis is a disease caused by Streptococcus sp., with clinical manifestations such as polyarthritis, septicemia, and meningitis. ?-hemolytic Streptococcus strains are known as one of the important pathogenic bacteria that cause serious illness. The bacteria enter their host through wounds, abrasions, food, or the immune system's decrease. This study aimed to determine the presence of ?-hemolytic Streptococcus spp in pigs raised on the people's farms in Punggul and Bongkasa Villages Abiansemal, Badung, Bali. As many as 47 isolates isolated from nasal mucosa swabs and tonsils of pigs were cultured on blood agar media, continued by planted them on Brain Heart Infussion (BHI) agar. Gram staining, catalase, and oxidaase tests were performed as biochemical tests to complete the identification of isolates. Finally, the bacitracin test was conducted on several isolates characterized with ?- hemolytic Streptococcus to differentiate Streptococcus pyogenes strains from others. The results show, 8 out of 47 (17%) samples tested are presumptive ?-hemolytic Streptococcus. Furthermore, the bacitracin test clarifies 3 out of 8 (37.5%) ?-hemolytic isolates are S. pyogenes, and the remaining 5 out of 8 (62.5%) are non-pyogenes ?-hemolytic Streptococcus.