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All Journal Medicina Buletin Veteriner Udayana E-Jurnal Medika Udayana Jurnal Veteriner Indonesia Medicus Veterinus ISM (Intisari Sains Medis) : Jurnal Kedokteran Jurnal Kedokteran: Media Informasi Ilmu Kedokteran dan Kesehatan Journal of Health Sciences and Medicine Journal of Global Pharma Technology Journal of Clinical Microbiology and Infectious Diseases

Komang Januartha Putra Pinatih

Departemen Mikrobiologi, Fakultas Kedokteran, Universitas Udayana, Denpasar, Bali, Indonesia

Author-ID : 2726193

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemistry Health Professions Immunology & microbiology Medicine & Pharmacology Public Health Veterinary

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Penentuan Marka Genetik Escherichia coli O157:H7 Asal Hewan dan Manusia dengan Metode Random Amplified Polymorphic DNA (GENETIC MARKERS IDENTIFICATION OF ESCHERICHIA COLI O157:H7 ORIGINATED FROM ANIMALS AND HUMAN BY USING RANDOM AMPLIFIED POLYMORPHIC DNA I Wayan Suardana; Dyah Ayu Widiasih; Komang Januartha Putra Pinatih
Jurnal Veteriner Vol 15 No 3 (2014)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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The use of random amplified polymorphic DNA (RAPD) as a method to identify a genetic markerof bacteria is widely used by researcher. This method is known as a simple, faster, and reliabletechnicque. This study is to find out the aplication of RAPD method in order to identify specific markersof E. coli O157:H7 as a zoonotic agent. The study began by cultivating of 20 isolates of E. coli O157:H7colected by previous study that consist of 2 isolates originated from cattle feces, 2 isolates originatedfrom beef, 2 isolates originated from chicken feces, 2 isolates originated from healthy human and 11isolates originated from unhealthy human (human with kidney failure). All isolates were confirmed byculturing on selective medium sorbitol MacConkey agar (SMAC). Confirmation were followed by testingon O157 latex aglutination, and finally by testing on H7 antiserum. RAPD method as molecularanalysis was performed using decamer primers mixture OPA-01, OPA-02, OPA-03, and OPA-04.Results of study showed both bands 1721 and 700 bp are specifically to differentiate of isolatesoriginated from cases of healthy and unhealthy human. On the other hand, bands with position 1721 bp,300 bp, and 250 bp indicate the isolates originated from unhealthy human, healthy human and chicken,respectively. Isolates from beef are characterized by both bands 1400 and 429 bp, and the isolates fromcattle feces are identified by band with position 342 bp. The specific bands are considered as markers inorder to know the source of E. coli O157:H7 fastly.

Sekuen Nukleotida Gene Shiga like toxin-2 dari Isolat Lokal Escherichia coli O157:H7 asal Hewan dan Manusia (NUCLEOTIDES SQUENCES OF SHIGA-LIKE TOXIN 2 GENES OF ESCHERICHIA COLI O157:H7 LOCAL ISOLATES ORIGINATED FROM ANIMALS AND HUMAN) I Wayan Suardana; Dyah Ayu Widiasih; Komang Januartha Putra Pinatih
Jurnal Veteriner Vol 18 No 1 (2017)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Animals/livestock, especially cattle, are known as the main reservoir of Escherichia coli O157: H7. As the only one of zoonotic E. coli, the pathogenicity of these bacteria is determined by its ability to produce one or more very potent cytotoxin known as Shiga-like toxin (Stx) or verocytotoxin, particularly of the Stx2 type that is closely related to the incidence of hemolytic uremic syndrome (HUS) in humans. This study analyzed the nucleotide sequences of stx2 gene between isolates from animals and humans in an effort to assess the potential zoonoses of the agent. The research activity was initiated by cultivating 20 isolates of E. coli O157:H7 collection based on result in the previous study i.e. 2 isolates originated from cattle feces, 2 isolates originated from beef, 2 isolates originated from chicken feces, 2 isolates originated from human feces, and 12 non-clinical isolates originated from human fecal who were suffering with renal failure. All isolates were confirmed on selective medium Sorbitol MacConkey Agar (SMAC) followed by testing on aglutination O157 latex test, and H7 antisera. Molecular analysis of stx2 gene covering open reading frame (ORF) of the stx2 gene was performed using the primer which was designed by researcher i.e. Stx2 (F)/Stx2 (R). The results showed, there were 2 isolates i.e. KL-48 (2) originated from human feces and SM-25 (1) originated from cattle feces were positive for carrying a stx2 gene, which was marked by the 1587 bp PCR product. Analysis of sequencing showed both isolates had identical to stx2 nucleotide squences with E. phaga 933 as well as E. coli ATCC 933. These results indicate the both local isolates are potential as zoonotic agents with clinical effects similar to E. phaga 933 and E. coli ATCC 43894. ABSTRAK Hewan ternak khususnya sapi, dikenal sebagai reservoir utama Escherichia coli O157:H7. Sebagai satu-satunya serotipe E. coli yang bersifat zoonosis, patogenitas bakteri ini ditentukan oleh kemampuannya untuk menghasilkan satu atau lebih cytotoxin yang sangat potensial yang dikenal dengan nama Shiga-like toxin (Stx) atau verocytotoxin, khususnya dari jenis Stx2 yang terkait erat dengan kejadian hemolytic uremic syndrome (HUS) pada manusia. Studi ini bertujuan menganalisis susunan nukleotida dari gen stx2 antara isolat asal hewan dan manusia dalam upaya mengkaji potensi zoonosis yang ditimbulkannya. Kegiatan penelitian diawali dengan kultivasi 20 isolat E. coli O157:H7 koleksi hasil penelitian sebelumnya dengan rincian dua isolat asal tinja sapi, dua isolat asal daging sapi, dua isolat asal tinja ayam, dua isolat asal tinja manusia non-klinis, dan 12 isolat asal tinja manusia klinis (asal penderita gagal ginjal). Isolat sebanyak 20 tersebut dikonfirmasi pada media selektif sorbitol MacConkey agar (SMAC) yang dilanjutkan dengan uji latex O157 aglutination test serta uji antiserum H7. Analisis molekuler komplit gen stx2 yang meliputi open reading frame (ORF) dari gen stx2 dilakukan menggunakan primer rancangan peneliti yaitu Stx2(F)/Stx2(R). Hasil penelitian menunjukkan bahwa ada dua isolat yaitu KL-48 (2) asal tinja manusia dan SM-25 (1) asal tinja sapi positif membawa gene stx2 yang ditandai dengan produk PCR 1587 bp. Analisis hasil sekuensing menunjukkan kedua isolat memiliki susunan gene stx2 yang identik dengan E. phaga 933 dan E. coli ATCC 43894. Hasil ini mengindikasikan kedua isolat lokal berpotensi sebagai agen zoonosis dengan efek klinis yang serupa dengan E. phaga 933 dan E. coli ATCC 43894.

Shiga Like Toxin 1 (STX-1) Detection From Escherichia coli O157:H7 Local Isolates I W. Suardana; K.J. P. Pinatih; D. A. Widiasih
Journal of Health Sciences and Medicine Vol 1 No 2 (2017): JHSM (September 2017)
Publisher : Institute for Research and Community Services Udayana University

Shiga-like toxin (Stx) otherwise known as verotoxin and verocytotoxin is a toxin produced by some strains of Escherichia coli particularly by strain O157: H7. This toxin is an AB5 toxin type which is known to have similarities with the toxins produced by Shigella dysenteriae . Stx from E. coli O157: H7 can be distinguished into stx 1 and stx 2. Stx1is usually associated with most outbreaks and detrimental sporadic cases of illness in humans. In this research, we observed the titer of Shiga toxin (Stx 1 / VT1) from local isolates isolated from cattle and human feces by using vero toxin Escherichia coli -reverse passive latex agglutination test (VTEC-RPLA) method. The results showed local isolates KL-48 (2) of human origin, SM-25 (1) of cattle feces origin and DS 21 (4) of beef origin positively produced VT1 2 units of titer, meanwhile the isolate SM 7 (1) was negative. Titer of toxin Stx1 produced from local isolates was known to be lower when compared to the control isolate ATCC 43894 with 8 units of titer.

Potential combinations of Platelet Rich Plasma (PRP) and chitosan in burn wound management: a review Gede Wara Samsarga; I Gusti Putu Hendra Sanjaya; Agus Roy Rusly Hariantana Hamid; Komang Januartha Putra Pinatih; Ni Made Ratih Purnama Dewi
Intisari Sains Medis Vol. 11 No. 2 (2020): (Available online: 1 August 2020)
Publisher : DiscoverSys Inc.

Background:Â The development of new modality for burn wound healing is necessary to reduce morbidity and mortality of burn injury. PRP (Platelet Rich Plasma) is one of potential modality for burn injury because of its ability to deliver various growth factors to initiate and accelerate tissue healing and regeneration. Clinical efficacy of PRP is frequently disturbed because inconsistency and inefficiency of growth factor release in PRP. To increase the effectiveness of PRP in wound healing and increase the bioavailability of growth factor, PRP can be combined with a carrier that can control continuous release of growth factors. Chitosan is one of biomaterial that potential to use because of its biodegradability characteristic and biological activity that important for burn wound healing and prevent the formation of scar tissue in a burn injury. Researches have shown that chitosan is an effective carrier to facilitate growth factor release continuously to burn wound.Aims: This review aims to review potential Combinations of PRP (Platelet Rich Plasma) & Chitosan in Burn WoundConclusion:Â With chitosan ability to facilitate burn wound healing and as an effective carrier, combination of PRP and chitosan is a promising modality for burn wound healing.

Prevalence of SCCmec Types I, II, III, and pvl gene among Methicillin-Resistant Staphylococcus aureus (MRSA) isolated from clinical specimens in Sanglah General Hospital I Gede Sathya Agastya; Agus Eka Darwinata; Komang Januartha Putra Pinatih; Ni Nengah Dwi Fatmawati
Intisari Sains Medis Vol. 13 No. 1 (2022): (Available Online : 1 April 2022)
Publisher : DiscoverSys Inc.

Background: Methicillin-Resistant Staphylococcus aureus (MRSA) is a big challenge for health services worldwide which causes infections both in healthcare and community. Healthcare-associated MRSA (HA-MRSA) strains are shown to be resistant to beta-lactam antibiotics and several non-beta lactam antibiotics. At the same time, the community-associated MRSA (CA-MRSA) tends to be resistant to beta-lactam antibiotics. MRSA carried staphylococcal cassette chromosome (SCCmec) types I, II, III, IV, and V. SCCmec types I, II, and III were predominantly found in HA-MRSA strain while SCCmec types IV and V predominantly found in CA-MRSA strains. Furthermore, the panton valentine leukocidine (pvl) gene is commonly found in CA-MRSA strains. Therefore, this study aimed to determine the prevalence of SCCmec types I, II, III, and pvl gene in MRSA isolated from clinical specimens in Sanglah General Hospital. Methods: This study was a cross-sectional descriptive study. MRSA was isolated from clinical specimens (sputum, wounds, tissue, blood, etc.) from January 2020 to July 2021 and identified by the Vitek 2 Compact (Biomerieux, France) at the Clinical Microbiology Laboratory of Sanglah Hospital. Prevalence of SCCmec and pvl gene using PCR. Data were analyzed using Microsoft Excel version 2010 for Windows. Results: Most of the specimens (69.56%) were wound. Seventeen (73.91%) out of 23 MRSA isolates were positive for the SCCmec III and pvl gene, while none was positive for the SCCmec I and SCCmec II. About 19 (82.60%) isolates were resistant to two or more non-beta-lactam antibiotics. Conclusions: The isolates of MRSA in this study were predominantly isolated from wound specimens, with the most prevalent genetic element being SCCmec III. In this study, although most MRSA isolates carried SCCmec III that suggested as HA-MRSA, however, most of the strains harbored the pvl gene. This interesting phenomenon needs to be further elucidated.

Contamination of Escherichia coli, Salmonella sp. and Vibrio sp. on ice cubes at food stalls in Karangasem Regency, Bali Province in 2021 I Gede Wikania Wira Wiguna; Agus Eka Darwinata; Komang Januartha Putra Pinatih; Ni Nengah Dwi Fatmawati
Intisari Sains Medis Vol. 13 No. 1 (2022): (Available Online : 1 April 2022)
Publisher : DiscoverSys Inc.

Background: Waterborne diseases spread through drinking water and food products made from drinking water, including cooking utensils that are washed using contaminated water. Ice cube is widely used by the people in Indonesia. There was no study regarding bacteria contamination on an ice cube in Karangasem Regency. This study aimed to determine the contamination of Escherichia coli, Salmonella sp., and Vibrio sp. on ice cubes at food stalls in Karangasem Regency in 2021. Methods: This study used a cross-sectional research design and simple purposive sampling as the sampling technique. Ice cubes were taken from 42 food stalls from eight sub-districts. Samples were cultured on Lactose Broth (LB) media for the Most Probable Number (MPN) test procedures and Eosin Methylene Blue Agar (EMBA) to identify the presence of E. coli, Salmonella Shigella Agar (SSA) for Salmonella sp., and Thiosulfate Citrate Bile Sucrose (TCBS) for Vibrio sp. and followed by gram staining. Data processing was carried out descriptively to determine the percentage of samples with positive contamination. Data were analyzed using Microsoft Excel version 2010 for Windows. Results: Based on the study results, it was found that all samples contained coliform bacteria through the MPN test (100.0%). In addition, 5 samples (11.9%) were found positive for E. coli. While the identification of Salmonella sp. and Vibrio sp was found no positive results. Conclusions: There was coliform bacteria contamination in the tested ice cube samples. In addition, E. coli was found in the sample; however, there were no Salmonella sp. and Vibrio sp. on ice cube samples.

Prevalensi cemaran patogen Salmonella pada daging babi yang dijual di Pasar Tradisional di Kota Denpasar I Gusti Agung Istri Gladys Elsyaningrat; Komang Januartha Putra Pinatih; Ni Nengah Dwi Fatmawati; Agus Eka Darwinata
Intisari Sains Medis Vol. 13 No. 2 (2022): (In Press : 1 August 2022)
Publisher : DiscoverSys Inc.

Background: Food contamination is a serious problem. Salmonella sp is one of the main bacteria causing gastrointestinal disease, especially in raw pork, due to the production and distribution process. Traditional markets have high risk of pathogenic microbial contamination. The purpose of this study was to detect contamination of Salmonella sp. on pork sold in traditional markets, particularly in Denpasar, Bali. Method: This research is a cross-sectional descriptive, with the samples coming from the porks sold by traditional market traders in every sub-district in Denpasar, Bali. The sampling technique used is the cluster purposive sampling. The procedure for identification of Salmonella sp. using culture and subculture examination on Salmonella Shigella Agar (SSA), simmon citrate agar media, and identification of the microscope. Results: Based on SSA culture results from 38 samples, 26 samples of pork showed colony growth, but only 10 samples with typical morphology of Salmonella sp. The 16 samples with atypical morphology were examined for SSA subculture, with 9 samples having clear black colonies. In 19 research samples that showed the results were suspected to be contaminated with Salmonella sp. continued to the Simmons citrate test, but only 8 samples were positive. The 19 samples were also examined under a microscope and all samples showed a rod image and red color indicating that the colonies in the sample were gram-negative bacilli which are typical of the morphology of Salmonella sp. Conclusion: Contamination of Salmonella was found in pork sold at Traditional Markets in Denpasar with a prevalence of 21%. Latar belakang: Kontaminasi makanan merupakan suatu masalah serius. Salmonella sp merupakan salah satu bakteri penyebab utama penyakit gastrointestinal, terutama pada daging babi mentah, dikarenakan pada proses produksi dan distribusi. Pasar tradisional merupakan tempat yang sangat rawan dan beresiko cukup tinggi terhadap cemaran mikroba patogen. Tujuan penelitian ini adalah mendeteksi ada tidaknya cemaran bakteri Salmonella sp. pada daging babi yang dijual di Pasar Tradisional, khususnya di Denpasar, Bali. Metode: Penelitian ini merupakan deskriptif potong lintang, dengan sampel penelitian berupa daging babi yang dijual oleh pedagang pasar tradisional di setiap kecamatan di Kota Denpasar, Bali. Teknik penentuan sampel yang digunakan yaitu teknik cluster purposive sampling. Prosedur identifikasi kontaminasi Salmonella sp. menggunakan pemeriksaan kultur serta subkultur pada Salmonella Shigella Agar (SSA), Kultur pada media simmon citrate agar, dan identifikasi mikroskop. Hasil: Berdasarkan hasil kultur SSA dari 38 sampel daging babi, 26 sampel daging babi menunjukan adanya pertumbuhan koloni, namun hanya 10 sampel dengan morfologi khas Salmonella sp. Pada 16 sampel dengan morfologi tidak khas dilakukan pemeriksaan subkultur SSA, dengan 9 sampel memiliki warna koloni hitam bening. Pada 19 sampel penelitian yang menunjukan hasil terduga terkontaminasi Salmonella sp. dilanjutkan ke uji Simmons citrate, namun hanya 8 sampel dengan hasil positif. Pada 19 sampel juga dilakukan pemeriksaan mikroskop dan seluruh sampel menunjukkan gambaran batang serta berwarna merah yang menandakan bahwa koloni pada sampel adalah basil gram negatif yang khas pada morfologi bakteri Salmonella sp. Kesimpulan: Terdapat cemaran patogen Salmonella sp. pada daging babi yang dijual di Pasar Tradisional di Denpasar dengan prevalensi sebesar 21%.

MOLECULAR ANALYSIS OF LACTIC ACID BACTERIA ISOLATE SR2 FROM BALI CATTLE RUMEN I Dewa Made Sukrama; Komang Januartha Putra Pinatih; I Wayan Suardana
Journal of Global Pharma Technology Volume 09 Issue 05
Publisher : Journal of Global Pharma Technology

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Bali cattle with its distinctively genetic characteristics, namely simple life or easy to adapt on unfavorable environmental condition was known as cattle pioneer. These characteristics were due to specific type of lactic acid bacteria (LAB). These facts indicate that the LAB identification with specific advantages was absolutely necessary for further research in order to prevent incorrect interpretation. Generally, molecular approach is the accurate method of identifying microorganism, and in this case, we used the analysis of the 16S rRNA gene. Research activities began with cultivation of SR2 isolates on specific media deMann Rogosa Sharpe (MRS) followed by Gram stain, catalase test, and formation of CO2 gas. Molecular analysis was performed by PCR of the 16S rRNA gene using primers 27F and 1492R. The PCR products were sequenced and nucleotides data were analyzed by Mega 5.2 Software tool. The results showed that phenotypical isolates of SR2 showed characteristics as LAB isolates, and the results of the 16S rRNA gene analysis showed that isolates SR2 included into Lactococcus lactis strain with a bootstrap value of 100.Keywords - Bali cattle, lactic acid bacteria, molecular analysis, rumen, SR2 isolate

IDENTIFICATION OF VIBRIO CHOLERAE, VIBRIO PARAHAEMOLYTICUS, AND VIBRIO VULNIFICUS BACTERIA IN BATIK SHELLS (PAPHIA UNDULATA) CAUGHT AT PENGAMBENGAN BEACH, JEMBRANA REGENCY Maharani, Ida Ayu Komang Ananta; Pinatih, Komang Januartha Putra; Darwinata, Agus Eka; Tarini, Ni Made Adi
E-Jurnal Medika Udayana Vol 13 No 9 (2024): E-Jurnal Medika Udayana
Publisher : Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/MU.2024.V13.i09.P12

Indonesia is a maritime country with vast seas and beaches equipped with natural riches stored in them. Global warming causes several changes in the ocean, including the increase in bacteria commonly found in the ocean, such as Vibrio sp. bacteria which can infect various animals in the ocean. Several species of bacteria Vibrio sp. that can infect humans are Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus which can cause significant complaints in the human digestive tract, wound infections, and even sepsis. This research aims to determine the bacterial contamination of Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus in batik shell (Paphia undulata) caught at Pengambengan Beach, Jembrana Regency. This research is a quantitative observational study with a cross sectional research approach and uses convenience sampling as a sampling technique. Based on several research stages starting from cultivating bacteria in APW liquid media to conducting tests on TSIA media, the results showed that from 70 samples, 16 samples were found to be positive for Vibrio sp. consisting of 11 positive samples for Vibrio vulnificus and five positive samples for Vibrio parahaemolyticus. The results of this study showed that 16 positive samples of Vibrio sp. bacteria were found from batik shell (Paphia undulata) caught at Pengambengan Beach, Jembrana Regency. Keywords: Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus

PREVALENSI KONTAMINASI BAKTERI Salmonella sp. DAN Shigella sp. PADA DAGING AYAM POTONG YANG DIJUAL OLEH PEDAGANG PASAR TRADISIONAL DI KOTA DENPASAR Surya Putra, I Gusti Agung Utama; Darwinata, Agus Eka; Putra Pinatih, Komang Januartha; Fatmawati, Ni Nengah Dwi
E-Jurnal Medika Udayana Vol 11 No 6 (2022): E-Jurnal Medika Udayana
Publisher : Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/MU.2022.V11.i06.P11

Salmonella sp. and Shigella sp. are bacterias that have become the most frequent cause of foodborne illness in developing countries. One of its contamination is chicken meat which is a daily staple food. The contamination occurs if the processing, storage, and distribution of chicken meat are improper. The aim of this study was to find the prevalence of contamination by Salmonella sp. and Shigella sp. on chicken meat sold by traditional market traders in Denpasar City. The type of research is cross-sectional research with descriptive observational. This study used the cluster sampling method. The total of samples is 24 samples and were taken from six traditional markets in each district of Denpasar City. Identification of Salmonella sp. and Shigella sp.Bacteria was done at the Integrated Biomedical Laboratory of Medical Faculty of Udayana with the culture method on Salmonella-Shigella agar media and Simmons Citrate culture. The total percentage contamination of Salmonella sp. was 58.33% (14/24), while Shigella sp. 20.83% (5/24). This bacterial contamination is caused by the lack of attention to hygiene in every chicken meat processing, storage, and distribution. The researcher suggests that further similar research be carried out using advanced identification methods to distinguish bacterial colonies in more detail and conduct a quantitative assessment of environmental contamination where chicken traders live.

Co-Authors Agus Eka Darwinata Agus Roy Rusly Hariantana Hamid Amy Yelly D. A. Widiasih Dhyana Ratmaja, I Gusti Agung Ngurah Dwi Lestari Dyah Ayu Widiasih Gede Wara Samsarga Samsarga I Dewa Ayu Rayna Nareswari Wikananda I Dewa Made Sukrama I G A Aristi Dhika Pranani I Gede Sathya Agastya I Gede Wikania Wira Wiguna I Gusti Agung Istri Gladys Elsyaningrat I Gusti Putu Hendra Sanjaya I Kadek Jaya Santika I Made Gustama Heryawan I Nyoman Arnatha I Putu Gede Septiawan Saputra I Wayan Suardana Ida Ayu Putu Putri Andari Ida Ayu Santhi Pertiwi Manuaba Ida Bagus Aditya Bhaskara Ida Bagus Putu Putrawan Ida Sri Iswari Kadek Adit Wiryadana Luh Putu Dea Sasmita Pralambari Made Agus Hendrayana Maharani, Ida Ayu Komang Ananta Mita Ekamelinda Nara Dhany, Henri Ni Luh Ranthi Kurniawathi Ni Made Adi Tarini Ni Made Ratih Purnama Dewi Ni Nengah Dwi Fatmawati Ni Nyoman Sri Budayanti Ni Putu Eka Umarista Apriliani Oktivia Chandra Mustika, Oktivia Chandra Putu Indri Widiani Rachmy Hamdiyati Rian Ka Praja Saraswati, I Gusti Ayu Agung Putri Indria Surya Putra, I Gusti Agung Utama Yosi Kusuma

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