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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).
Acinetobacter baumannii Is an opportunistic pathogen as an MDRO especially on intensive ward Suranadi, I Wayan; Dwi Fatmawati, Ni Nengah; Aryabiantara, I Wayan; Sinardja, Cynthia Dewi; Saputra, Darmawan Jaya
Bali Journal of Anesthesiology Vol 3, No 2 (2019)
Publisher : DiscoverSys Inc.

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (169.797 KB) | DOI: 10.15562/bjoa.v3i2.199

Abstract

Acinetobacter baumannii is an opportunistic bacterial pathogen that is associated with hospital acquired infections and is a major cause of nosocomial infections especially in intensive spaces; this is becoming increasingly a widespread concern in various hospitals around the world. Acinetobacter baumannii, which is resistant to many antibiotics, is now recognized as clinically very important. Reports suggest that the spread of A. baumannii in the hospital environment led to an increase in nosocomial outbreaks associated with high mortality rates. However, many other Acinetobacter spp. can also cause nosocomial infections. This review focuses on the role of Acinetobacter spp. as nosocomial pathogens, resistance patterns and epidemiology.
IMMUNE RESPONSE AND COST ANALYSIS OF INTRADERMAL RABIES VACCINATION FOR POST-EXPOSURE PROPHYLAXIS REGIMEN IN HUMAN Budayanti, N. S.; Susilawathi, N. M.; Darwinata, A. E.; Dwija, I. B. P.; Fatmawati, N. D.; Wirasandhi, K.; Subrata, K.; Susilarini, N. K.; Wignall, F. S.; Sudewi, A. A. R.; Mahardika, and G. N. K.
BALI MEDICAL JOURNAL Vol 3 No 1 (2014)
Publisher : BALI MEDICAL JOURNAL

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (229.703 KB)

Abstract

Background The outbreak of rabies in human in Bali-Indonesia is causing an extraordinary pressure for the government in providing adequate doses of anti-rabies vaccine for post-exposure prophylaxis (PEP). Here, we directly compare the immune response and benefit of the intradermal (ID) protocol for rabies vaccine delivery with the intramuscular (IM) route. Methods: Sixty health workers who were willing to participate in this study have been randomly selected and grouped into ID, IM, and control groups, each with 20 volunteers. The Thai Red Cross ID- and Zangreb IM-protocols have been applied to the respective group. The sera of the volunteers were collected at day 0, week 1, week 3, week 4, month 3, month 6, month 9, and month 12 after the first vaccination. Anti-rabies virus IgG was detected using PlateliaTM Rabies II Kit (Bio-Rad). Results: Anti-rabies IgG could be detected in the ID-group at one week. The ID-vaccine delivery induced a slightly higher maximum antibody titer compared to IM, though not statistically significant (p>0.05). ID vaccination caused less adverse reactions and produces longer lasting protective immune response. Cost minimization analysis (CMA) on the provincial and national PEP data in 2009-2011 shows that the ID-delivery will reduce the total cost for a completed regimen by USD 28.5, and would have saved the Indonesian government budget approximately USD 3.6 and 4.3 million for complete regimens in Bali and Indonesia, respectively. Conclusion: The ID administration of anti-rabies vaccine induces a similar immune response compared to that of intramuscular injection. It also produces longer lasting protective immune response. It offers additional advantages of potential net cost savings as well as decreasing the pressure on vaccine availability due to the high number of dog bite cases.
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.
THE EXISTENCE OF VIBRIO CHOLERAE IN INDONESIA: FROM ENVIRONMENTAL TO CLINICAL ASPECTS (A CONCISE REVIEW) Rian Ka Praja; Anggita Ratri Pusporini; Reny Rosalina; I Wayan Muda Suta Arta; I Dewa Made Sukrama; Ni Nengah Dwi Fatmawati
Jurnal PPI Dunia Vol 4 No 1 (2021)
Publisher : OISAA

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.52162/jie.2021.004.01.1

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.
DETECTION METALLO-BETA-LACTAMASE GENE IMP-1 AND IMP-2 OF Pseudomonas aeruginosa CLINICAL ISOLATES IN SANGLAH HOSPITAL BALI Ni Made Adi Tarini; Ni Nengah Dwi Fatmawati; I Putu Bayu Mayura
International Journal of Biosciences and Biotechnology Vol 3 No 1 (2015)
Publisher : Central Laboratory for Genetic Resource and Molecular Biology, Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (412.06 KB)

Abstract

Pseudomonas aeruginosa is a pathogen frequently found as an agent of Hospital Acquired infections. This bacterium is very easy to be resistant to several types of antibiotics through various mechanisms. Carbapenem such as Imipenem and Meropenem is a potential option for the therapy of this bacterium, but unfortunately P. aeruginosa has ability in hydrolyzing these antibiotics through enzyme metallo-?-lactamases (MBLs). Recently, IMP and VIM, MBLs enzyme group are reported common from various countries, but no data is reported for these enzymes in Indonesia especially in Bali. In fact, the resistant data of P. aeruginosa against carbapenem group antibiotics such as meropenem and imipenem is quite high in Sanglah General Hospital in 2014 was 35% and 45% respectively. Therefore, the aim of this study was to detect IMP-1 and IMP-2 genes of MDR P. aeruginosa, which are phenotypically resistant to the antibiotic Imipenem and Meropenem disks based on CLSI standards in Clinical Microbiology Laboratory, Sanglah General Hospital, Denpasar, Bali. Eighty-six isolates were isolated from sputum (25 / 29.1%), wound (25 / 29.1%), urine (15 / 17.4%),endotracheal Tube (11 / 12.8), pus (6/7% ), blood (3 / 3.5%) and tissue (1 / 1.1%). In this study, all isolates were subjected to PCR for detection of IMP-1 and IMP-2. The result showed that 9 isolates were positive IMP-1 gene (10.5%), but there was no isolate positive for IMP-2 gene. The result was similar with that of the other countries, especially for the gene IMP-1. Detection and molecular characterization of MBL-producing P. aeruginosa strains are very important for infection control purposes. Currently, this study is still continued for detection of another MBL genes.
MULTIPLEX PCR FOR DETECTION OF CAPSULAR POLYSACCHARIDES TYPES OF Streptococcus pneumoniae CLINICAL ISOLATES IN BALI Ni Nengah Dwi Fatmawati; Ni Made Adi Tarini; I Putu Bayu Mayura
International Journal of Biosciences and Biotechnology Vol 2 No 2 (2015)
Publisher : Central Laboratory for Genetic Resource and Molecular Biology, Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

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Abstract

Streptococcus pneumoniae is causative agent of non-invasive and Invasive Pneumococcal Diseases(IPD). One of the major virulence factors is capsular polysaccharides (CPS). The CPS is known as thepneumococcal vaccine component. Several types of S. pneumoniae CPS are dominant in Indonesiasuch as types 6, 23, 15, 33 and 12 in West Nusa Tenggara, type 7F in Jakarta, and types 6A/B dan15B/C in Central Java. No data is reported from Bali related to S.pneumoniae CPS typing. Therefore,the aim of this study was to determine CPS types of S. pneumoniae isolates in Clinical MicrobiologyLaboratory, Sanglah General Hospital, Denpasar, Bali by using Multiplex PCR. Twenty-one isolatesthat were isolated from blood (11/52.4%), sputum (5/23.8%), and other clinical specimens (5/23.8%)were included in this study. Identification of S. pneumoniae was based on optochin test and presenceof pneumolysin gene (ply). Uniplex PCR was conducted to determine capsular type of each isolates,and then continued with Multiplex PCR 1 and 2, which used in-house positive controls. All isolateswere positive for the presence of ply, confirming the isolates were S. pneumoniae. Moreover, thisstudy showed that type 19F was the predominant type (7 isolates (66.7%)); 2 isolates (9.5%) werepositive for each type 23F and also for type 6A/B; and, there was only 1 isolate (4.8%) for each type7F and 15B/C. Total of 8 isolates (38.1%) were found to be nontypeable isolates. Multiplex PCR wassuccessfully identified different types of CPS. Development of Multiplex PCR could help indiagnosing and identifying capsular type of S. pneumoniae simultaneously.
Isolasi dan Idetifikasi Bakteri aam Laktat Penghasil Bakteriosin dari Urutan, Sosis Fermentasi Tradisional Bali Ni Made Sri Dwijastuti; I Nengah Sujaya; Ni Nengah Dwi Fatmawati
Metamorfosa: Journal of Biological Sciences Vol 8 No 1 (2021)
Publisher : Prodi Magister Ilmu Biologi, Fakultas MIPA, Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/metamorfosa.2021.v08.i01.p08

Abstract

This study aimed to isolate and identify lactic acid bacteria (LAB) that could produce bacteriocins from urutan. Urutan is a Balinese sausage that is traditionally produced and naturally fermented. Urutans used in this study were obtained from several traders in the Baturiti, Mengwi, and Tegallalang region, Bali. More than 300 LAB colonies were isolated and underwent inhibition test against the growth of the indicator bacteria. Crude bacteriocins from two isolates, namely J2 and J6, showed inhibition zones against the growth of Lactobacillus plantarum E12.1 and Escherichia coli ATCC 8739. The J6 isolate also inhibited the growth of Enterococcus faecalis ATCC 29212. Both J2 and J6 were gram-positive, bacilli shaped, and showed negative catalase test. Based on the results of the 16S rDNA gene amplification, the two isolates were identified as Lactobacillus plantarum. Keywords: Lactic Acid Bacteria, Bacteriocin, Lactobacillus plantarum, Urutan
Metode Deteksi Carbapenem Resistant Enterobacteriaceae Ni Wayan Eka Putri Gayatri Kastawa; Yan Ramona; N.N. Dwi Fatmawati.
Metamorfosa: Journal of Biological Sciences Vol 7 No 1 (2020)
Publisher : Prodi Magister Ilmu Biologi, Fakultas MIPA, Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/metamorfosa.2020.v07.i01.p07

Abstract

Carbapenem Resistant Enterobacteriaceae (CRE) is a group of Enterobacteriaceae resistant to Carbapenem antibiotics (Imipenem, Ertapenem, Meropenem, Doripenem). One species of Enterobacteriaceae frequently showing resistant characteristic to Carbapenem is Klebsiella pneumoniae. Early detection methods on the CRE existence (phenotypically or genetically) are urgently needed. The methods should be able to detect carbapenemases or genes encoding these types of enzymes. In the recent years, phenotypic characteristics can be done by applying methods, such as Modified Hodge Test (MHT), Carba Nordmann-Poirel (Carba NP), or Modified Carbapenem Inactivation Method (MCIM). To detect the presence of genes encoding production of carbapenemases, Polymerase Chain Reaction (PCR) is the most reliable method available in the recent years. Keywords: Carbapenem Resistant Enterobacteriaceae (CRE), Phenotype, Genothype, Modified Carbapenem Inactivation Method (MCIM), Modified Hodge Test (MHT), Carba Nordmann-Poirel, Polymerase Chain Reaction (PCR).
KARAKTERISTIK FENOTIF ISOLAT KLINIK Escherichia coli O157:H7 PADA MEDIA SORBITOL MAC CONKEY AGAR (SMAC) Wahyu Hidayati; I Gede Rai Maya Temaja; Ni Nengah Dwi Fatmawati
Journal of Agricultural Science and Biotechnology Vol 7 No 1 (2018)
Publisher : Journal of Agricultural Science and Biotechnology

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Abstract

Phenotypic Characteristic of Escherichia coli O157: H7 Clinical Isolates on MacConkey Agar with Sorbitol (SMAC)Escherichia coli is a Gram-negative, rod-shaped bacterium, and commonly has flagella. Most of E. coli strains are normal flora in digestive tract of human, but some serotypes are pathogen for human and animal. One of pathogenic E. coli strain that causes a severe infection in humans (hemorrhagic colitis and hemolytic uremic syndrome) is known as Enterohemorrhagic Escherichia coli (EHEC) with cattle as their natural reservoir. E. coli O157:H7 is the most important and pathogenic serotype of EHEC which responsible for outbreak of Hemorrhagic Colitis and Hemolytic Uremia Syndrome. Accurate and cheap method detection of E. coli O157:H7 is needed to help early detection diagnosis and therapy. Therefore, the aim of this study is investigate phenotypic characteristics of E. coli O157:H7 isolated from clinical specimens using MacConkey Agar with Sorbitol (SMAC) media. Three E. coli clinical isolates from Sanglah General Hospital showed colorless non-sorbitol fermenting colony in SMAC media as phenotypic characteristic of E. coli O17:H7, therefore SMAC may beused on are of confirmation methods for E. coli O157:H7 detection from clinical isolats.
Co-Authors Adhy Candra, I Kadek Bayu Agus Eka Darwinata Andi Yasmon Anggita Ratri Pusporini Aviana, Felicia Bagus Ngurah Putu Arhana Bryan Setiawan Budiman Bela Cynthia Dewi Sinardja Dave Gerald Oenarta Dea Antariksa Dharmika, Ida Ayu Gde Wahyudevi Dhyana Ratmaja, I Gusti Agung Ngurah Dwijastuti, Ni Made Sri F. S. Wignall Fera Ibrahim Gema Zakharian GNR Suwardana I Dewa Agung Gede Meisha Dhanam I Dewa Ayu Made Dian Lestari I Dewa Made Sukrama I G. A. Ngurah Aswin Panji Sanjaya I G. R. M. TEMAJA I Gde Raka Widiana I Gede Agus Darsana Palgunadi I Gede Gita Sastrawan I Gede Pradnya Wisnu Murthi I Gede Raka Adhyatma I Gede Sathya Agastya I Gede Wikania Wira Wiguna I Gusti Agung Dyah Ambarawati I Gusti Agung Istri Gladys Elsyaningrat I Gusti Ayu Agung Praharsini I Gusti Ayu Janadewi I Gusti Ngurah Kade Mahardika I Gusti Ngurah Krishna Priyaka I Kadek Jaya Santika I Ketut Suastika I Made Sathya Vijayananda I Made Sutha Saskara I Nengah Sujaya I Putu Bayu Mayura I Putu Bayu Mayura I Wayan Agus Gede Manik Saputra I Wayan Aryabiantara, I Wayan I Wayan Gustawan I Wayan Muda Suta Arta I Wayan Suranadi I. B. P. Dwija I.D.P.K. Pratiwi Ichlazul Ma’ruf Ida Ayu Andhira Dewi Suarisavitra Ida Bagus Gede Adiguna Wibawa Ida Sri Iswari Indramawan Setyojatmiko K. Subrata K. Wirasandhi Kadek Anggie Wigundwipayana Kadek Tresna Yuwana Ketut Tuti Parwati Komang Aditya Arya Prayoga KOMANG AYU NOCIANITRI Komang Januartha Putra Pinatih Kusuma, Anak Agung Ngurah Jaya Luh Gede Melia Puspita Sari Made Agus Hendrayana Made Gede Dwi Lingga Utama Made Widianantara Marvin Giantoro N. K. Susilarini N.W.A. Utami Ni Kadek Seri Mahayanti Ni Luh Made Rasmawati Ni Made Adi Tarini Ni Made Mertaniasih Ni Made Susilawathi Ni Nyoman Sri Budayanti Ni Putu Wirantari Ni Wayan Eka Putri Gayatri Kastawa NMRP Dewi Prisela Zharaswati Prisillia Brigitta Putu Agung Satvika Pradnyadevi Putu Ayu Utami Prajawaty Putu Yoska Arya Harindana Rachmy Hamdiyati Raka-Sudewi A. A. Reny Rosalina Rian Ka Praja Saputra, Darmawan Jaya Saraswati, I Gusti Ayu Agung Putri Indria Sonia Elvira Salim Suastika, Arresta Vitasatria Surya Putra, I Gusti Agung Utama Wahyu Hidayati Wahyu Hidayati Wahyu Hidayati Yan Ramona