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Identifikasi Daging Babi Menggunakan Metode PCR-RFLP Gen Cytochrome b dan PCR Primer Spesifik Gen Amelogenin Yuny Erwanto; Sugiyono Sugiyono; Abdul Rohman; Mohammad Zainal Abidin; Dwi Ariyani
agriTECH Vol 32, No 4 (2012)
Publisher : Faculty of Agricultural Technology, Universitas Gadjah Mada, Yogyakarta, Indonesia

A polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) and species specific PCR methods had been applied for identifying pork in mixture of meat. Pork sample in various levels (1, 3, 5 and 10%) was prepared in mixture with beef, chicken and mutton. The primary CYTb1 and CYTb2 were designed in the mitochondrial cytochrome b b (cytochrome b) gene and PCR successfully amplified fragments of 359 bp. To distinguish pig species existence, the amplified PCR products of mitochondrial DNA were cut by BseDI restriction enzyme. The result showed that pig mitochondrial DNA was cut into 131 and 228 bp fragments. A polymerase chain reaction (PCR) method based on the nucleotide sequence variation in the amelogenin gene has been chosen for the specific identification of pork DNAs in mixture meat. The primers designed generated specific fragments of 353 and 312 bp length for pork. The specificity of the primary designed was tested on 4 animal species including pig, cattle, chicken and goat species. Analysis of experimental mixture meat demonstrated that 1% of raw pork tissues could be detected using PCR-RFLP with BseDI restriction enzyme but detection using species-specific PCR showed the cross reactivity to beef, chicken and mutton. The cytochrome b PCR-RFLP species identification assay yielded excellent results for identification of pig species. PCR-RFLP is a potentially reliable technique for detection of the existence of pork in animal food product for Halal authentication. ABSTRAKPenelitian ini dilakukan untuk mengaplikasikan metode deteksi daging babi dalam campuan daging dengan sapi, kambing dan ayam melalui PCR-RFLP dan PCR dengan primer spesifik untuk babi. Level kontaminasi daging babi dibuat sebesar 1, 3, 5 dan 10% dari total daging dalam campuran. Metode PCR-RFLP menggunakan sepasang primer yaitu gen cytochrome b dari mitokondria yang menghasilkan fragmen DNA sebesar 359 bp. Untuk mengetahui ada tidaknya kontaminasi babi dalam adonan daging tersebut diaplikasikan enzim restriksi BseDI yang dapat memotong DNA dari gen cytochrome b babi. Hasil penelitian menunjukkan bahwa gen cytochrome b dari babi dapat terpotong menjadi dua fragmen yaitu sebesar 228 bp dan 131 bp. Untuk desain primer spesifik digunakan gen amelogenin yang mempunyai sekuen yang berbeda diantara ke empat spesies uji yaitu babi, sapi, ayam dan kambing. Primer spesifik didesain pada panjang fragmen sebesar 353 dan 312 bp. Hasil penelitian menunjukkan bahwa kontaminasi daging babi sebesar 1% masih dapat terdeteksi dengan metode PCR-RFLP tetapi pengujian dengan primer spesifik yang ditujukan hanya untuk deteksi DNA babi masih menunjukkan reaksi silang dengan spesies hewan lain yaitu sapi, kambing dan ayam. Pengujian dengan PCR-RLP pada gen cytochrome b menghasilkan hasil yang lebih baik dan jelas untuk pengujian kontaminasi babi dibandingkan dengan PCR dengan primer spesifik. Metode PCR-RFLP merupakan metode yang potensial untuk analisis deteksi keberadaan unsur babi pada produk olahan pangan khususnya untuk deteksi status kehalalan.

Gambaran Histopatologi Otak Tikus Akibat Injeksi Trimetyltin sebagai Model Penyakit Alzheimer Yuli Purwandari Kristianingrum; Sitarina Widyarini Sitarina Widyarini; Kurniasih Kurniasih; Bambang Sutrisno Bambang Sutrisno; Charles Rangga Tabbu Charles Rangga Tabbu Charles Rangga Tabbu; Sugiyono Sugiyono
Jurnal Sain Veteriner Vol 34, No 1 (2016): Juni
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Trimethyltin chloride (TMT) is an organotin compound which neurotoxic at limbus system and hippocampus in human and animal. Pathology changes that caused by the induction of TMT is a neurodegenerative disorder such as nerve cell death and cognitive impairment. This study was aimed to observe brain pathology induced by TMT with multiple doses for 14, 21 and 28 days after treatment. Twenty seven of Wistar rats, at 2 months of age with weight ranging between 200-300 grams were used and divided randomly into 3 groups (n=9). Group I were injected by trimetyiltin with a dose of 6 mg / kg, group II were injected bytrimetyltin with a dose of 8 mg / kg and group III as control without injection. Observation of brain pathology was done by euthanasia on day 14, 21 and 28 after treatment, three rats each. Cortex and hippocampus of the brainwere observed using Hematoxilin and Eosin staining (HE). All of the research procedure was done with the approval and supervision of Animal Ethics Committee LPPT UGM No. 300/KEC-LPPT/VII/2015. The observation of histopathology of the brain's neuron cells injected by trimetyltin dose of 6 mg/kg and 8 mg/kg body weight was showed increasing cell death of brain neurons in the cortex and hippocampus compared to the control group. The highest cell death was on day 14 in the hippocampus and cortex cerebral on day 21after TMT injection. The neuron cell death characterized by the shrink of brain neurons as well as colored eosinophilic cytoplasm. One way ANOVA statistical analysis showed a significant difference number of neurons cell deathbetween control and treatment groups. Based on this research, it can be concluded that the trimetyltin injection dose of 6 mg / kg and 8 mg / kg of body weight caused neuron cell death in the brain rats from fourteen day aftertreatment, especially in the hippocampus and cortex.

Gambaran Histopatologi Otak Tikus Akibat Injeksi Trimetyltin sebagai Model Penyakit Alzheimer Yuli Purwandari Kristianingrum; Sitarina Widyarini Sitarina Widyarini; Kurniasih Kurniasih; Bambang Sutrisno Bambang Sutrisno; Charles Rangga Tabbu Charles Rangga Tabbu Charles Rangga Tabbu; Sugiyono Sugiyono
Jurnal Sain Veteriner Vol 34, No 1 (2016): Juni
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Trimethyltin chloride (TMT) is an organotin compound which neurotoxic at limbus system and hippocampus in human and animal. Pathology changes that caused by the induction of TMT is a neurodegenerative disorder such as nerve cell death and cognitive impairment. This study was aimed to observe brain pathology induced by TMT with multiple doses for 14, 21 and 28 days after treatment. Twenty seven of Wistar rats, at 2 months of age with weight ranging between 200-300 grams were used and divided randomly into 3 groups (n=9). Group I were injected by trimetyiltin with a dose of 6 mg / kg, group II were injected bytrimetyltin with a dose of 8 mg / kg and group III as control without injection. Observation of brain pathology was done by euthanasia on day 14, 21 and 28 after treatment, three rats each. Cortex and hippocampus of the brainwere observed using Hematoxilin and Eosin staining (HE). All of the research procedure was done with the approval and supervision of Animal Ethics Committee LPPT UGM No. 300/KEC-LPPT/VII/2015. The observation of histopathology of the brain's neuron cells injected by trimetyltin dose of 6 mg/kg and 8 mg/kg body weight was showed increasing cell death of brain neurons in the cortex and hippocampus compared to the control group. The highest cell death was on day 14 in the hippocampus and cortex cerebral on day 21after TMT injection. The neuron cell death characterized by the shrink of brain neurons as well as colored eosinophilic cytoplasm. One way ANOVA statistical analysis showed a significant difference number of neurons cell deathbetween control and treatment groups. Based on this research, it can be concluded that the trimetyltin injection dose of 6 mg / kg and 8 mg / kg of body weight caused neuron cell death in the brain rats from fourteen day aftertreatment, especially in the hippocampus and cortex.

Immunodiagnosis Infeksi Aeromonas hydrophila pada Ikan Yuli Purwandari Kristianingrum; Sitarina Widyarini; Kurniasih Kurniasih; Bambang Sutrisno; Charles Rangga Tabbu; Sugiyono Sugiyono
Jurnal Sain Veteriner Vol 36, No 1 (2018): Juni
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Aeromonas hydrophila causes a disease that often infects fish and is known as Motile Aeromonas Septicaemia (MAS), Hemorrhagi Septisemia, Ulcer disease or Red-Sore disease. The aims of this study were to develop polyclonal antibody of Aeromonas hydrophila in the rabbits to confirm the diagnosis of Aeromonas hydrophila in the fish by immunohistochemistry staining method. Preparation of polyclonal antibodies was performed on the rabbits used to Aeromonas hydrophila bacteria that have been tested biochemically by intravenous and intraperitoneal injection. Doses of Aeromonas hydrophila bacteria were 109 CPU/ml of 0.5 ml at first injection, 1 ml at second injection, 2 ml at thirth injection and 3 ml at fourth injection. Blood serum collection was performed at week 5 after injection from an ear and intracardial vein. The result of antibody titer was 28 = 1024 which measured by tube test. Furthermore, polyclonal antibody was used to immunohistochemistry staining with 400x dilution. The results of the staining showed that an immunopositive reaction in the liver, skin,lien, gill, kidney, and heart of fish to Aeromonas hydrophila antibody. The research conclution was polyclonal antibody from rabbit can be used to accurately confirm the diagnosis of Aeromonas hydrophila based on antigen and antibody reaction.

Studi In-Vivo Ekstrak Daun Teh Hijau (Camellia Sinensis) sebagai Alternatif anti Bakteri Eschericia Coli pada Ayam Broiler Bambang Sutrisno; R. Wasito; Kurniasih Kurniasih; Sitarina Widyarini; Yuli Purwandari Kristianingrum; Sugiyono Sugiyono
Jurnal Sain Veteriner Vol 37, No 2 (2019): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

The prevalence of colibasillosis in chicken farms in Indonesia is very high, treatment using antibiotics is experiencing resistance, so it is necessary to look for alternatives to antibacterial. The study was aimed to determine the antibacterial effect of green tea leaf extract on broiler chickens infected with Eschericia coli by looking at the score of macroscopic lesions strengthened by histopathological examination, heterophile examination, plasma protein and fibrinogen. The research used 20 day old broilers (DOC) which were randomly divided into 4 groups, group A group B, group C and group D, each consisting of 5 DOC broilers. While maintaining ND and Gumboro vaccines on schedule like maintenance in general. At the age of 21 days all broilers in each group began to be treated as controls (Group A) without infecting E. coli and were not given 0,1g/ml water extract of green tea leaves (Camillia sinensis). Group B, intratracheal-infected broilers with local strains of E.coli were 108 cells / ml according to 0,5 Mc Farland standard, and were not given green tea leaf extract. Group C, broilers infected by intratracheal with local strains of E. coli 108 cells / ml by 0,5 Mc Farland standard, and given to drink green tea leaf extract (Camillia sinensis) 0,1 g/ml and group D, broilers were given drinking green tea leaf extract (Camillia sinensis) 0,1g/ml. During the treatment all of chickens were given food and drink ad libitum. Fourteen days after infection of E.coli, 5 chickens in each group were collected to collect blood for heterophyll, total plasma protein (TPP) and fibrinogen. And then were euthanasied with Mg SO4 saturated solution intravenously injection and necropsied for gross and histpathological examination. Analysis of blood tests results were used one way of anova (SPSS version 22 program), whereas for gross and histopathological examination with descriptive analysis. The results showed that the gross examination and histopathological organs of brolier infected with E. coli without being given a green tea extract experienced airsacculitis, pericarditis, perihepatitis and peritonitis, whereas broilers infected with E. coli and given green tea extract does not indicate the presence of inflammation. Examination of heterophile counts and blood fibrinogen levels had shown a difference (P <0.05), in broilers infected with E. coli and given green tea extracts had lower amounts of hetrophils and fibrinogen levels. While blood TPP levels were not significantly different (P> 0.05). The conclusion can be drawn, that the study of in vivo green tea extract (Camelia sinensis) 0,1g/ml has the potential to inhibit the infection of Eschericia coli bacteria in broiler chickens.

Gangguan Pertumbuhan Organ Limfoid Ayam Broiler yang Menderita Omfalitis Bambang Sutrisno; R Wasito; Sitarina Widyarini; Yuli Purwandari Kristianingrum; Sugiyono .
Jurnal Sain Veteriner Vol 39, No 3 (2021): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.60465

Penelitian ini bertujuan untuk melihat gangguan pertumbuhan jaringan limfoid primer dan sekunder yang menderita omphalitis dengan pemeriksaan histopatologi diwarnai dengan pewarnaan hematoksilin-eosin rutin dan pewarnaan imunohistokimia streptavidin biotin terhadap interleukin-10 (IL-10) pada ayam muda. Ayam umur 24 hari (DOC) broiler digunakan dan dikumpulkan dari tempat penetasan yang sama di Jawa Tengah di Indonesia. Semua 24 DOC dibagi menjadi dua kelompok yang masing-masing terdiri dari 12 DOC (Grup A) dan 12 DOC omphalitic (Grup B). Semua DOC dirawat di kandang yang berbeda, diberi makan dan diminum di libitum. Pada hari ke 3, 6 dan 9, empat ekor ayam dari masing-masing kelompok ditimbang untuk kemudian dinekropsi. Timus, bursa fabricius dan limpa dikumpulkan dan ditimbang. Semua jaringan diproses secara histopatologi dengan pewarnaan hematoksilin-eosin rutin dan pewarnaan biotin streptavidin imunohistokimia imunopatologi. Data indeks berat limpa, bursa Fabricius dan tymus dianalisis menggunakan program statistik IBM SPSS versi 22. Hasil penelitian menunjukkan bahwa indeks bobot limpa, bursa fabricius dan timus ayam omphalitic (kelompok B) lebih rendah dibandingkan dengan indeks bobot ayam sehat (kelompok A). Indeks berat timus berbeda nyata (P <0, 05). Lesi histopatologis pada organ limfoid diamati pada semua ayam di Grup B. Lesi ditandai dengan penipisan dan nekrosis limfosit. Ayam dari Grup A tidak mengalami perubahan pada organ limfoid. Biotin streptavidin immunostaining dengan ekspresi antibodi policlonal anti IL-10 pada bursa Fabricius pada ayam omphalitic (Grup B) memiliki IL-10 yang sangat sedikit jika dibandingkan dengan ayam sehat (Grup A). Kesimpulan, omfalitis menyebabkan penurunan indeks berat badan yang signifikan dan gangguan pertumbuhan organ limfoid yang ditandai dengan deplesi dan nekrosis limfosit.

Gambaran Mikroskopis Berbagai Organ Merpati yang Menunjukkan Gejala Tortikolis dan Identifikasi Virus Penyakit Tetelo Berdasarkan Uji Agar Gel Presipitasi Risang Aji Dewandaru; Ferdinand Prayogo Cahyo Santoso; Sugiyono Sugiyono; Michael Haryadi Wibowo
Jurnal Veteriner Vol 21 No 4 (2020)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

This study aimed to determine the microscopic conditions of organs of pigeons that suffered from torticollis and identify the cause of disease in torticollis pigeons. Three pigeons showed torticollis symptoms were obtained as sample and marked as pigeon A, B and C, respectively. Isolation of pathogen with inoculation into embryonated pigeon eggs obtained from parents with no history of vaccination and not indicated ND. Identification of ND with agar gel precipitation (AGP) test. Observation of microscopical changes with histopathologic preparation using hematoxylin and eosin staining. Histopathological examination showed that pigeons was done severe neuritis vagus, trakheitis, pneumonia, air sacculitis, hepatitis, pankreatitis, nefritis, jejunoileitis, ileocolitis and orchitis. Perivascular cuffing found in brain. Degenerative changes found in the hepar and ren. Cardiac severe necrotic lesion, and depletion in white pulp area of spleen. Proventricular tissue showed flattening of mucosal epithelium, congestion lesions found in pulmonary tissue. The results of slow hemagglutination test of pigeon egg allantoic fluid, which tested for hemagglutination (HA) showed positive HA result with titers varying between 2, 32, and 64. Serological identification carried out with the AGP test on all culture samples against ND antiserum showing positive results of ND virus. Based on these dara, it can be concluded that the pigeons withsymptoms of torticollis is caused by ND virus.

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