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Metagenomic Sequencing of P30-gene-positive Using MinION Sequencer for Apicomplexan Species Identification Dela Ria Nesti; Ahmad Baidlowi; Lucky Ronald Runtuwene; Ari Indrawati; Wayan Tunas Artama; Yutaka Suzuki
Proceeding of International Conference on Science, Health, And Technology 2021: Proceeding of the 2nd International Conference Health, Science And Technology (ICOHETECH)
Publisher : LPPM Universitas Duta Bangsa Surakarta

Oxford Nanopore Technologies MinION sequencer is a recent technology of portable, real-time, single molecule long-read DNA/RNA sequencing which has the potential to analyze clinical and environmental samples in the field. Here, we describe the result of metagenomic sequencing from samples with P30-gene-positive using MinION sequencer. One hundred cattle blood samples which were confirmed positive of blood parasites using blood smear method were collected from Diseases Investigation Centre (DIC), Yogyakarta province. After DNA extraction, each sample was amplified by conventional PCR using primers targeting the P30 gene. PCR products were visualized under UV illumination after electrophoresis on 1.5% TBE/agarose gels and stained with SYBR Safe™ DNA gel stain. Twenty-five samples showed the desired 400-bp DNA fragment, which was inferred as P30 gene positive. Four whole cattle blood samples with the highest DNA concentration were sequenced separately using MinION sequence for 24 hours. Sequencing data was then analyzed using bioinformatics tools. Three samples resulted in low sequencing yield and could not be analyzed, but we were able to identify Theileria orientalis as the major pathogen from one sample. We were also able to construct 95.8% of its genome. To our knowledge, this is the first publication of a parasite genome reconstruction from metagenomic sample using MinION sequencer in Indonesia.

KAJIAN LEPTOSPIROSIS PADA SAPI POTONG DI DAERAH ALIRAN SUNGAI PROGO DAERAH ISTIMEWA YOGYAKARTA (The Study of Bovine Leptospirosis in Progo Watershed, Yogyakarta) Guntari Titik Mulyani; Bambang Sumiarto; Wayan Tunas Artama; Sri Hartati; Juwari J; Sugiwinarsih S; Henricus Roby Cahya Putra; Estu Widodo
Jurnal Kedokteran Hewan Vol 10, No 1 (2016): March
Publisher : Universitas Syiah Kuala

The purpose of this study was to determine the prevalence, serovar, and risk factors of leptospirosis on cattle in Progo watershed. A total of 330 cattle samples were clinically examined and blood was collected for Leptospira examination using microscopic agglutination test (MAT). Results of MAT were used as dependent variable (Y), while breeders, cattle, shed, and feed factors were used as the independent variable (X). Univariate analysis, bivariate analysis with chi square, odds ratio, and multivariate with logistic regression were implemented to analyze the data. The results showed that all cattle were cinically health, 13.03% (43/330) cattle were positively leptospirosis which 37 out of 193 (9.17%) cattle belongs to the farmer. Mostly, leptospirosis cases were caused by hardjo serovar (38.0%). Risk factors influencing the prevalence of leptospirosis were caused by some factors such as the presence of rats in cage (OR 2.7), the distance of waste disposal which should be less than approximately 5 meters (OR 2.1), feed concentrates, grass and rice straw (OR 0.4), grazing (OR 0.2), and concentrate feed and rice straw (OR 0.1). It can be concluded that leptospirosis infection on cattle in Progo watershed is 13.03% (at livestock level) and 19.17% (at farm levels). Factors of rat in the cage and the distance of waste disposal (5 m) increase the risk of leptospirosis cases. Factors of feed combine with concentrate and rice straw as well as grazing reduced the risk of leptospirosis in cattle, but the addition of grass in concentrate feed and rice straw provides a greater risk of leptospirosis infection compare to concentrate feed and rice straw alone.Key words: beef cattle, leptospirosis, prevalence, risk factors

MOLECULAR EXPRESSION OF WINGLESS-TYPE MMTV INTEGRATION SITE FAMILY MEMBER 4 GENE USING Escherichia coli BL21 Agung Janika Sitasiwi; Wayan Tunas Artama; Agung Budiyanto; Edy Dharmana
Jurnal Kedokteran Hewan Vol 11, No 1 (2017): March
Publisher : Universitas Syiah Kuala

This research was conducted to find out the Wnt4 recombinant proteins which expressed by Escherichia coli (E. coli) BL21 carrying the recombinant DNA wnt4 (E. coli transformation). Research materials were E. coli BL21 transformation and E. coli BL21 non-transformation (negative control). The expression of recombinant protein was conducted by culturing E. coli for 24 hours in Luria-Bertani (LB) media with isopropyl β-D-1-thiogalactopyranoside (IPTG) induction. Recombinant protein was isolated by sonication of pellet bacteria. Protein analysis performed by 15% sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that recombinant protein with a molecular weight of 33 kDa has been expressed by E. coli BL21 transformation successfully.

DETECTION OF SAG1 AND BAG1 Toxoplasma gondii DNA PROBES LABELLED WITH DIGOXIGENIN-11-dUTP Ida Ayu Pasti Apsari; Wayan Tunas Artama; Sumartono s; I Made Damriyasa
Jurnal Kedokteran Hewan Vol 7, No 1 (2013): March
Publisher : Universitas Syiah Kuala

The objective of this research was to detect a minimum concentration of the probes that could be used for dot blot hybridization analysis. The method required labeled DNA probes. In this study a non-radioactive label of Digoxigenin-11-dUTP was used for labeling the Sag1 and the Bag1 of Toxoplasma gondii DNA probe. Labeling method for the probes was done according to the random primed labeling technique. The result showed that 0.67 pg/µl Sag1 probe and 0.58 pg/µl Bag1 probe could be detected by anti-Dig-antibody. It could be concluded that 0.67 pg/µl Sag1 probe and 0.58 pg/µl Bag1 probe could be used to diagnose toxoplasmosis by dot blot hybridization method.

KAJIAN LEPTOSPIROSIS PADA SAPI POTONG DI DAERAH ALIRAN SUNGAI PROGO DAERAH ISTIMEWA YOGYAKARTA (The Study of Bovine Leptospirosis in Progo Watershed, Yogyakarta) Guntari Titik Mulyani; Bambang Sumiarto; Wayan Tunas Artama; Sri Hartati; Juwari J; Sugiwinarsih S; Henricus Roby Cahya Putra; Estu Widodo
Jurnal Kedokteran Hewan Vol 10, No 1 (2016): March
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v10i1.3374

The purpose of this study was to determine the prevalence, serovar, and risk factors of leptospirosis on cattle in Progo watershed. A total of 330 cattle samples were clinically examined and blood was collected for Leptospira examination using microscopic agglutination test (MAT). Results of MAT were used as dependent variable (Y), while breeders, cattle, shed, and feed factors were used as the independent variable (X). Univariate analysis, bivariate analysis with chi square, odds ratio, and multivariate with logistic regression were implemented to analyze the data. The results showed that all cattle were cinically health, 13.03% (43/330) cattle were positively leptospirosis which 37 out of 193 (9.17%) cattle belongs to the farmer. Mostly, leptospirosis cases were caused by hardjo serovar (38.0%). Risk factors influencing the prevalence of leptospirosis were caused by some factors such as the presence of rats in cage (OR 2.7), the distance of waste disposal which should be less than approximately 5 meters (OR 2.1), feed concentrates, grass and rice straw (OR 0.4), grazing (OR 0.2), and concentrate feed and rice straw (OR 0.1). It can be concluded that leptospirosis infection on cattle in Progo watershed is 13.03% (at livestock level) and 19.17% (at farm levels). Factors of rat in the cage and the distance of waste disposal (5 m) increase the risk of leptospirosis cases. Factors of feed combine with concentrate and rice straw as well as grazing reduced the risk of leptospirosis in cattle, but the addition of grass in concentrate feed and rice straw provides a greater risk of leptospirosis infection compare to concentrate feed and rice straw alone.Key words: beef cattle, leptospirosis, prevalence, risk factors

MOLECULAR EXPRESSION OF WINGLESS-TYPE MMTV INTEGRATION SITE FAMILY MEMBER 4 GENE USING Escherichia coli BL21 Agung Janika Sitasiwi; Wayan Tunas Artama; Agung Budiyanto; Edy Dharmana
Jurnal Kedokteran Hewan Vol 11, No 1 (2017): March
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v11i1.5891

This research was conducted to find out the Wnt4 recombinant proteins which expressed by Escherichia coli (E. coli) BL21 carrying the recombinant DNA wnt4 (E. coli transformation). Research materials were E. coli BL21 transformation and E. coli BL21 non-transformation (negative control). The expression of recombinant protein was conducted by culturing E. coli for 24 hours in Luria-Bertani (LB) media with isopropyl β-D-1-thiogalactopyranoside (IPTG) induction. Recombinant protein was isolated by sonication of pellet bacteria. Protein analysis performed by 15% sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that recombinant protein with a molecular weight of 33 kDa has been expressed by E. coli BL21 transformation successfully.

DETECTION OF SAG1 AND BAG1 Toxoplasma gondii DNA PROBES LABELLED WITH DIGOXIGENIN-11-dUTP Ida Ayu Pasti Apsari; Wayan Tunas Artama; Sumartono s; I Made Damriyasa
Jurnal Kedokteran Hewan Vol 7, No 1 (2013): March
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v7i1.579

The objective of this research was to detect a minimum concentration of the probes that could be used for dot blot hybridization analysis. The method required labeled DNA probes. In this study a non-radioactive label of Digoxigenin-11-dUTP was used for labeling the Sag1 and the Bag1 of Toxoplasma gondii DNA probe. Labeling method for the probes was done according to the random primed labeling technique. The result showed that 0.67 pg/µl Sag1 probe and 0.58 pg/µl Bag1 probe could be detected by anti-Dig-antibody. It could be concluded that 0.67 pg/µl Sag1 probe and 0.58 pg/µl Bag1 probe could be used to diagnose toxoplasmosis by dot blot hybridization method.

AN EVALUATION STUDY OF ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) USING RECOMBINANT PROTEIN GRA1 FOR DETECTION OF IgG ANTIBODIES AGAINTS TOXOPLASMA GONDII INFECTIONS Muflikhah, Nina Difla; Artama, Wayan Tunas
Indonesian Journal of Tropical and Infectious Disease Vol. 6 No. 5 (2017)
Publisher : Institute of Topical Disease Universitas Airlangga

Toxoplasmosis is an infectious disease caused by Toxoplasma gondii, an intracellular protozoan parasite that live inside the cells of the reticulo endothelial and parenchymal cells of human and animals (mammals and birds). Some cases of toxoplasmosis usually have no symptoms, but in any cases caused severe symptoms, such as hydrocephalus, microcephalus, intracranial calcification, retinal damage, brain abscess, mental retardation, lymphadenopathy, and others. Its severe symptoms usually showed a long time after first exposure, except symptoms showed by congenital transmission caused by infected mother. Early diagnosis is important to prevent the illness but methods for toxoplasmosis screening are still too expensive for developing country. Enzyme-linked immunosorbent assay (ELISA) allow the testing of a large number samples within short time frame and based on antibody or antigen detection. This study aimed to know the sensitivity and specificity of recombinat protein GRA1 as antigen using ELISA methods. We tested the sensitivity and spesificity of GRA1 protein as antigen in ELISA methods to diagnose toxoplasmosis and compared with ELISA Kit Commercial. Reliable laboratory testing is important to detect Toxoplasma gondii infection, and focused to improving the low cost and easy-to-use diagnostic instrument. Seventy sera collected and tested using both indirect ELISA, commercial ELISA kit and GRA1 protein coated as antigen. Fourty eight and fifty one samples showed positive IgG antibody result of ELISA-GRA1 and ELISA kit. Negative sample tested by ELISA-GRA1 was 22 samples and 19 sample tested by ELISA Kit. The sensitivity and specificity of GRA1-based on ELISA were 100% and 86.36%, positive prediction value (ppv) was 94.11%. These data indicate that the recombinant protein GRA1 is a highly immunogenic protein in human toxoplasmosis and become a promising marker for the screening of toxoplasmosis.

Sacrificers' Preferences on Selection and Procurement of Sacrificial Animals for Eid al-Adha Celebration Ibrahim, Alek; Budisatria, I Gede Suparta; Artama, Wayan Tunas; Widayanti, Rini; Atmoko, Bayu Andri
ANIMAL PRODUCTION Vol. 24 No. 1 (2022)
Publisher : Faculty of Animal Science, Jenderal Soedirman University in associate with the Animal Scientist Society of Indonesia (ISPI) and the Indonesian Association of Nutrition and Feed Science (AINI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20884/1.jap.2022.24.1.99

Eid al-Adha is one of the biggest religious celebrations in Indonesia. This study was conducted to determine the profile and activities of sacrificers (shohibul qurban) in selecting and procuring sacrificial animals for the Eid al-Adha celebration. This study was conducted by in-depth interviews with a total of 54 sacrificers in Sleman Regency, Yogyakarta, during the Eid al-Adha celebration in 2019. The data were analyzed using descriptive analysis (number and percentage). This study indicated that most sacrificers were classified as productive age (45.35 years), high school graduates (46.30%), and self-employed workers (50.00%) with an income of IDR 3,357,896 per month. The sacrificial animals were cattle, sheep, and goats with an average body weight of 253.75 kg, 27.80 kg, and 32.80 kg, and the prices of IDR 20,939,962; IDR 2,906,250; and IDR 2,420,000, respectively. The sacrificial animals were mainly purchased from the livestock seller or intermediary seller (38.89%). There were livestock-price differences in the different marketplaces (44.44%) and between Eid al-Adha and the daily period (61.11%). This study concluded that the cattle preferred animal types for group sacrifices, and the sheep had preferred animal types for individual sacrifice. Cattle are usually sacrificed with a joint purchase between sacrificers. Each reason for selecting and procuring the sacrificial animal types was influenced by different motivations, such as financial, animal handling, and local wisdom factors.

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