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All Journal Jurnal Sain Veteriner Jurnal Veteriner BERITA BIOLOGI Jurnal Kedokteran Hewan The International Journal of Tropical Veterinary and Biomedical Research Prosiding Seminar Nasional Unimus

Wisnu Nurcahyo

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Author-ID : 2760486

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

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Title

Penggunaan Protein Membran Stadium Bradizoit Toxoplasma gondii untuk Mendiagnosis Toksoplasmosis dengan Metode Intradermal Test Muhammad Hanafiah; Wisnu Nurcahyo; Dwinna Aliza; Teuku Fadrial Karmil
Jurnal Veteriner Vol 14 No 2 (2013)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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A research was conducted to find out an alternative diagnose in detecting toxoplasmosis in livestock/animal using intradermal test from protein membrane of T. gondii bradizoite stage. Local isolate ofmembrane T. gondii bradizoite stage was used in the research. Ten of domestic sheep with the age of ± 1year and 10 mice strain Balb/c with the age ± 2 month were used in this research. The reaction ofhypersensitivity on the skin post protein membrane bradyzoite injection was indicated by the process ofskin thickening. The diameter skin thickening was measured using cutimeter, in which diameter e” 10 mmindicated positive diagnose. The result showed that optimal dosage of membran protein bradyzoite thatcould be applied to detect toxoplasmosis in livestock and animal using intradermal test were 0,6 ml and0,2 ml for sheep and mice respectively. The sensitivity and specificity level of antigen use (protein membrane)of T. gondii bradizoite stage from local isolate to diagnose toxoplasmosis in mice using intradermal testwere: 85.0 % and 66.6 % respectively, while in sheep the sensitivity and specificity level were 85.0 % and66.6 % respectively. It can be concluded that intradermal test was appropriate to be implemented fordetecting toxoplasmosis in sheep and mice induced with tachyzoite T. gondii.

Molecular Detection of Toxoplasmosis Using Specific Primers P30, B1, and rDNA Wisnu Nurcahyo; Joko Prastowo; Ana Sahara
Jurnal Veteriner Vol 13 No 1 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Study in order to develop molecular techniques using specific primers for the early diagnosis oftoxoplasmosis have been conducted. Detection of Toxoplasma gondii genome was performed usingpolymerase chain reaction (PCR) technique. The primers used in this study were rDNA, P30, and B1. ThePCR products were further run using gel electrophoresis (gel 1.5% – 2.0%) and the band was documented.Toxoplasma was detected at 500 bp and 600 bp using primer P30 and B1, respectively. Whereas usingprimer rDNA no band was observed. It was assumed that primer rDNA was not sensitive since the targetamplification was 88 bp.

Daya Larvasida Ekstrak Daun Muda Kedondong Hutan Terhadap Haemonchus contortus Secara In-vitro (LARVICIDAL EFFECT OF SPONDIAS PINNATA LEAF EXTRACT AGAINST HAEMONCHUS CONTORTUS IN VITRO) I Gusti Komang Oka Wirawan; Wisnu Nurcahyo; Joko Prastowo; Kurniasih Kurniasih
Jurnal Veteriner Vol 18 No 2 (2017)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

This study aimed to asses effective concentration of condensed tannins in the extract of Lannea grandis leaves (EDMKH) as larvacide anti-helminths on Haemonchus contortus. Experimental design used in this study was completely randomised design. The treatments group were administrated with 2.5%, 3.5% , and 4.5% EDMKH in the 0.2 g/mL solution, respectively. Moreover aqueduct was used for the negative control group, while 0.055% albendazole was applied to the positive control group. Variables measured were percentage of developed and undeveloped L1 in the incubation media. The data were the statistically analysed following General Linear Model format for completely randomised design. The software used in the analysis was SPSS v. 23.0 (IBM-SPSS 2015). Results showed that among the EDMKH treatments, the lowest larvacidal effect was recorded for 2.5% EDMKH treatment compared with 3.5% and 4.5% EDMKH. However, the value achieved by 2.5% EDMKH treatment was significantly higher (P<0.05) than that of negative control treatment. The highest larvacidal effect (100%) was shown by 4.5% EDMKH treatment which was not significantly different (P>0.05) from the positive control treatment. ABSTRAK Penelitian ini bertujuan untuk mengetahui konsentrasi efektif tanin kondensasi di dalam Ekstrak Daun Muda Kedondong Hutan (EDMKH) sebagai antelmintik yang bersifat larvasida terhadap Haemonchus contortus (H. contortus) secara in-vitro. Rancangan penelitian yang digunakan dalam penelitiam ini adalah Rancangan Acak Lengkap (RAL), dengan lima perlakuan yaitu tiga perlakuan EDMKH konsentrasi: 2,5%, 3,5%, 4,5% dari 0,2 g/mL larutan tersedia, satu kelompok kontrol (-) menggunakan air suling, satu kelompok kontrol (+) menggunakan albendazole konsentrasi 0,055%. Variabel yang diukur dan dianalisis: jumlah persentase L1 yang berkembang dan tidak berkembang dalam kelompok rendaman. Data persentase daya larvasida H. contortus dari konsentrasi tanin kondensasi EDMKH secara in-vitro dianalisis menggunakan analisis varian mengikuti prosedur Rancangan Acak Lengkap (RAL) menggunakan SPSS Versi 23(IBM-SPSS 2015). Hasil analisis statistika menunjukkan bahwa persentase larvasida EDMKH 2,5% nyata lebih rendah dibandingkan 3,5%, 4,5% dan albendazole 0,055% (P<0,05) tetapi lebih tinggi dari kontrol negatif. Konsentrasi efektif EDMKH terhadap larva H. contortus adalah konsentrasi 4,5%, memberikan efektivitas larvasida 100%, dan tidak berbeda nyata dengan kontrol positif (P>0,05).

Pengembangan Antibodi Poliklonal dari Stadium Oosista, Sporosista, dan Sporozoit Eimeria tenella (THE DEVELOPMENT OF POLYCLONAL ANTOBODY FROM EIMERIA TENELLA OOCYST, SPOROCYST, AND SPOROZOITE STADIUM) Galuh Tresnani; Joko Prastowo; Wisnu Nurcahyo; Budi Setiadi Daryono
Jurnal Veteriner Vol 14 No 2 (2013)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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The research on developing diagnostic method, vaccine, and drugs for coccidiosis has been focused onthe finding of the immunogenic molecule in Eimeria. The identification of this agent will need the antibodywhich can recognize the biomolecule in the antigen. Antibody that has been developed for this purposeshould be analyzed first, and one of the simple methods for analyzing this antibody is through dot blotanalysis. The objective of this research was to analyze the polyclonal antibody which developed from theoocyst, sporocyst, and sporozoite of E. tenella using dot blot analysis. The antigen for this polyclonalantibody was made from each of the E. tenella stadium by sonication. Fifteen mice, divided into 3 groups,were then injected subcutaneously with each antigen. The sera from these mice were then collected, analyzedby using ELISA and then it will be used for the dot blot analysis. The research result showed that thepolyclonal antibody which has been developed in mice from each antigen can react with the antigen itself.From this result it can be concluded that the developing of this antibody is successful and it can be used forfurther research in immunoproteomic.

Kekerabatan Genetik Caplak Rhiphicephalus (Boophilus) microplus Asal IndonesiaBerdasarkan Sekuen Internal Transcribed Spacer-2 (GENETIC RELATIONSHIP INDONESIAN RHIPHICEPHALUS (BOOPHILUS) MICROPLUS TICK BASED ON INTERNAL TRANSCRIBED SPACER-2 SEQUENSE ) Ana Sahara; Joko Prastowo; Rini Widayanti; Kurniasih .; Wisnu Nurcahyo
Jurnal Veteriner Vol 16 No 3 (2015)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Rhiphicephalus (Boophilus) microplus is important obligatory blood feeding ectoparasites transmittingmany different viral, bacterial and protozoan and plays a role as a vector of Babesia sp., The leria sp. andAnaplasma sp. in cattle. The accuracy in identifying and distinguishing interspecies and intraspeciesdiversity among parasites is needed to understand the epidemiology, biology and capacity as a vector.Variations in the DNA base sequence of the internal transcribed spacer region2 (ITS 2) has been used asa molecular marker for identification in an effort to determine phylogenetic relationships. The aim of thisstudy was to determine the ITS 2 gene nucleotide sequence of R. microplus, which was expected to beuseful for accurate identification the parasite diversity and phylogenetic relationship among many differentspecies. DNA amplification was conducted using BOO2 forward dan BOO2 reverse primers. The DNAsamples containing ITS2 region fragment of 1099 nt were derived from the nucleotide sequence multiplealignments of R.microplus and other ticks genes obtained from Gene bank using Clustal W software, andthen analyzed using the MEGA program version 6. Genetic distances based on nucleotide sequence weredetermined with Kimura 2-parameter method producing the smallest genetic distance of 0 % and 1.2 %.Construction of phylogenetic trees using the Neighbor joining method showed that ticks from variousregions in Indonesia was species complex which have a closer with R.microplus isolates from India, Laos,South Africa, China and Australia R.australis origin.

Produksi dan Isolasi Protein Membran Stadium Bradizoit Toxoplasma gondii : Suatu Usaha untuk Mendapatkan Material Diagnostik dalam Mendiagnosa Toksoplasmo Muhammad Hanafiah; Wisnu Nurcahyo; Mufti Kamaruddin; Fadrial Karmil
Jurnal Veteriner Vol 10 No 3 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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A study was conducted to isolate the membrane protein of Toxoplasma gondii at bradyzoid stage forthe development of intradermal diagnostic test in livestock infected by the parasite. Toxoplasma wasinitially collected from meat of goat serologically positive to the parasite. The infected meat was then fedinto uninfected cat to obtain oocyst. The oocyst was inoculated into the stomach of mice to produce tachyzoitwhich in turn produce cyst in tissue known as bradyzoit . The membrane protein was then isolated from thebradyzoit. The protein was analysed by sodium dodecyl sulfate electrophoresis (SDS-PAGE). The dataobtained were presented descriptively. The protein concentration isolated from each mouse infected at thedose of 1x107 oocysts was 11.91 mg. Two protein bands specific for bradyzoit were identified at 97.72 kDaand 67.60 kDa.

Profil Protein Stadium Sporozoit Eimeria tenella Isolat Yogyakarta Melalui Analisis Protein SDS-PAGE (PROTEIN PROFILE OF THE SPOROZOITE OF Eimeria tenella ISOLATES FROM YOGYAKARTA USING SDS-PAGE PROTEIN ANALYSIS) Galuh Tresnani; Joko Prastowo; Wisnu Nurcahyo; Budi Setiadi Daryono
Jurnal Veteriner Vol 13 No 2 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Coccidiosis is one of the important diseases in poultry industry. In Indonesia the morbidity of thedisease is between 80 to 90%. A rapid and prompt diagnosis would be one of the essential steps ineradication and control of the disease. The objective of this study is to determine the protein profile ofsporozoite of Eimeria tenella isolated in Yogyakarta using sodium dodecyl sulphate polyacrilamide gelelectrophoresis (SDS-PAGE) protein analysis. Protein analysis was performed in 12% polyacrilamide geland further electrophoresis at 100 volts and over-staining with Coomasie brilliant blue. The resultsshowed that the sporozoite of E. tenella isolated in Yogyakarta contained five proteins with molecularweights of 15, 26, 32, 80, and 91 kDa, respectively.

Antigen Ekskretori-Sekretori Cacing Jantung (Dirofilaria immitis) Jantan dan Betina yang Berpotensi Sebagai Marka Diagnosis (EXCRETORY-SECRETORY ANTIGENS OF MALE AND FEMALE HEART WORMS (DIROFILARIA IMMITIS) WHICH POTENTIALLY AS A DIAGNOSTIC MARKER) I Gusti Made Krisna Erawan; Ida Tjahajati; Wisnu Nurcahyo; Widya Asmara
Jurnal Veteriner Vol 16 No 4 (2015)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Heart worm (Dirofilaria immitis) is the causative agent of a serious parasitic disease in dogs.Dirofilariasis is generally diagnosed by microfilariae examination and specific antigen testing. Microfilariaeexamination has low sensitivity due to occult infections. The available antigen test at this time is able todetect circulating antigens secreted by adult female worms only. The aim of the present study was toidentify male (MES) and female (FES) heart worms excretory-secretory antigens which have the potentialas a diagnostic targets. Identification of antigen was done by sodium dodecyl sulphate polyacrylamide gelelectrophoresis (SDS-PAGE) and Western Blotting analysis. The results of this study indicated that therewere differences between the MES and the FES profiles. The results showed 12 bands in MES (14–118kDa) and 18 bands in FES (10–205 kDa). Protein with a molecular weight of 59 kDa has the potential asdiagnostic markers of dirofilariasis.

Gambaran Histopatologi Toksoplasmosis pada Kucing Peliharaan (HISTOPATHOLOGICAL FEATURES OF TOXOPLASMOSIS IN DOMESTIC CAT) Muhammad Hanafiah; Wisnu Nurcahyo; Joko Prastowo; Sri Hartati
Jurnal Veteriner Vol 18 No 1 (2017)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Study of histopathological changes of domestic cat organs which were serologically positive toxoplasmosis and laboratory infected which Toxoplasma have been undertaken. Histological section is prepared from organs including brain, liver, lung, kidney, duodenum, jejunum, ileum, and spleen then stained using hematoxylin and eosin (HE) and observed under microscope for histopathological changes. The results showed that in the serologically positive animals cell proliferation, infiltration of leucocyte and macrophage cells were observed in the ileum, whilst infiltration of eosinophil and leucocyte was seen in the kidney and liver. However, in other organ such as duodenum, jejunum, and spleen there were no changes observed. In cat experimentally infected with Toxoplasma, the infiltration of eosinophil cells were observed in the ileum and lung, while other organs such as kidney, liver, brain, jejunum, duodenum, and spleen showed no infiltration of inflammation cells. In conclusion, based on the results seropositive cat, showed proliferation of epithelial cells, leucocyte cells, and macrophage cells in the ileum, while in the lung, kidney, and liver showed infiltration of eosinophil and leucocyte. No infiltration of inflammation cells were observed in the brain, jejunum, duodenum, and spleen. ABSTRAK Penelitian mengenai histopatologi beberapa organ kucing peliharaan yang positif Toxoplasma baik secara serologi maupun yang diinfeksikan telah dilakukan. Tujuan penelitian adalah untuk mengetahui perubahan histopatologi pada organ kucing yang positif Toxoplasma. Data hasil pemeriksaan histopatologi yang terdapat pada preparat jaringan masing-masing organ dianalisis secara deskriptif dengan melihat gambaran perubahan histopatologi pada organ otak, hati, paru, ginjal, duodenum, jejenum, ileum, dan limpa. Hasil penelitian menunjukkan bahwa dengan menggunakan metode histopatologi organ kucing yang positif toksoplasmosis secara serologi teramati adanya proliferasi sel epitel, infiltrasi sel-sel leukosit dan makrofag pada ileum, ginjal, dan hati terlihat adanya infiltrasi eosinofil dan juga infiltrasi leukosit, sedangkan organ yang lain seperti jejenum, duodenum, dan limpa tidak teramati perubahan pada jaringan yang diperiksa. Sementara pada kucing yang dinfeksikan Toxoplasma, ileum dan paru teramati adanya infiltrasi sel-sel eosinofil, sedangkan organ lainnya seperti ginjal, hati, otak, jejenum, duodenum, dan limpa tidak teramati adanya infiltrasi sel-sel radang. Simpulan yang dapat ditarik adalah pada organ kucing yang positif toksoplasmosis teramati adanya proliferasi sel epitel, infiltrasi sel-sel leukosit, dan makrofag pada ileum, paru, ginjal dan hati teramati adanya infiltrasi eosinofil dan juga infiltrasi leukosit, sedangkan organ-organ lainnya seperti otak, jejenum, duodenum dan limpa tidak terlihat adanya infiltrasi sel-sel radang.

Co-Authors Ana Sahara Ana Sahara BUDI SETIADI DARYONO Dikry N Shatrie Dwi Priyo Widodo Dwinna Aliza Erdiansyah Rahmi Erdiansyah Rahmi Fadrial Karmil Galuh Tresnani I Gusti Komang Oka Wirawan I Gusti Made Krisna Erawan Ida Tjahajati Joko Prastowo Kurniasih . Kurniasih Imamudin Kurniasih Kurniasih Luluk Ria Rakhma M. Hanafiah M.Hanafiah . Muchlison Anis Mufti Kamaruddin Muhammad Hanafiah Muhammad Hanafiah R. Wasito . Rini Widayanti Sri Hartati Sumartono . Teuku Fadrial Karmil Triyanto Triyanto Widya Asmara

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