Isolasi, Identifikasi, dan Gambaran Makroskopis dan Mikroskopis pada Berbagai Organ Ayam Layer dengan Gejala Avian Influenza

Shakhra Zaha Fairuza, Prof. Dr. drh. Michael Haryadi Wibowo, M.P.

2025 | Skripsi | KEDOKTERAN HEWAN

Avian Influenza (AI) merupakan penyakit yang menyerang unggas, disebabkan oleh virus Influenza tipe A dan merupakan penyakit menular. Penyakit ini merugikan secara ekonomi, menyebabkan penurunan produksi telur, bahkan kematian massal. Penelitian ini bertujuan untuk isolasi dan identifikasi virus AI sebagai penyebab kematian massal pada sebuah peternakan di Desa Tirtomartani, Sleman, Yogyakarta menggunakan Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), telur ayam berembrio (TAB), serta mengetahui perubahan patologi anatomi secara makroskopis dan mikroskopis. Dua ekor ayam dengan gejala anoreksia, letargi, kesulitan bernafas dinekropsi. Trakea, limpa, dan paru-paru diambil untuk dilakukan uji molekuler menggunakan RT-PCR dengan primer target gen H5 (583 bp) dan H9 (712 bp). Jengger, hepar, jantung, trakea, ginjal, usus, paru, limpa, ovarium, dan kloaka diambil untuk dilakukan analisis secara histopatologis. Isolasi pada TAB non-SPF dilakukan dengan menginokulasikan organ trakea dan pool paru-paru lien. Cairan alantois dipanen, diuji hemaglutinasi (HA), hemaglutinasi inhibisi (HI), serta RT-PCR dengan primer target gen H5 (583 bp) dan H9 (712 bp). Hasil pengamatan makroskopis menunjukkan lesi perdarahan subkutan otot dada, kekeruhan mesenterium, diskolorisasi dan pembengkakan hepar, perdarahan lemak abdominal, deformitas calon telur, dan perdarahan ovarium. Perubahan mikroskopis menunjukkan kongesti jengger, kongesti vena sentralis dan sinusoid disertai nekrosis hepatosit, nekrosis otot jantung dengan beberapa serabut otot mengalami vakuolisasi, erosi epitel mukosa disertai infiltrasi sel radang pada usus, deplesi limfosit limpa, dan perdarahan ovarium. Hasil uji molekuler two step RT-PCR sampel organ trakea dan pool paru-paru lien menunjukkan hasil negatif. Hasil uji hemaglutinasi alantois sampel trakea menunjukkan hasil positif. Hasil hemaglutinasi inhibisi menggunakan antiserum AI dan Newcastle Disease (ND) menunjukkan hasil negatif, dan uji molekuler two step RT-PCR menggunakan primer target gen H5 (583 bp) dan H9 (712 bp) sampel alantois hasil pasase menunjukkan hasil negatif. Berdasarkan pengamatan patologi anatomi, perubahan berupa kongesti jengger, perdarahan otot dada, nekrosis hepar, nekrosis jantung, radang usus, dan perdarahan ovarium menunjukkan bahwa ayam terindikasi terinfeksi AI meskipun virus AI tidak terisolasi dan teridentifikasi.

Avian Influenza (AI) is a contagious disease affecting poultry, caused by the Influenza A virus. This disease leads to economic losses due to decreased egg production and mass mortality. This study aimed to isolate and identify the AI virus as the cause of mass mortality on a farm in Tirtomartani Village, Sleman, Yogyakarta, using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and embryonated chicken eggs (ECEs), as well as to determine the gross and microscopic anatomical pathology changes. Two chickens presenting with anorexia, lethargy, and respiratory distress were necropsied. The trachea, spleen, and lungs were sampled for molecular testing using RT-PCR with primers targeting the H5 (583 bp) and H9 (712 bp) genes. The comb, liver, heart, trachea, kidneys, intestines, lungs, spleen, ovary, and cloaca were collected for histopathological analysis. Virus isolation was conducted in non-SPF ECEs by inoculating tracheal organs and a lung-spleen pool. The harvested allantoic fluid was tested using Hemagglutination (HA), Hemagglutination Inhibition (HI), and RT-PCR with primers for the H5 (583 bp) and H9 (712 bp) genes. Gross examination revealed subcutaneous hemorrhages in the pectoral muscle, mesenteric cloudiness, liver discoloration and swelling, abdominal fat hemorrhages, misshapen ova, and ovarian hemorrhages. Microscopic changes included comb congestion; central venous and sinusoidal congestion with hepatocyte necrosis; cardiac muscle necrosis with vacuolization of some muscle fibers; mucosal epithelial erosion with inflammatory cell infiltration in the intestine; splenic lymphocyte depletion; and ovarian hemorrhages. The two-step RT-PCR results from the tracheal and pooled lung-spleen organ samples were negative. The Hemagglutination (HA) test on the allantoic fluid from the tracheal sample was positive. However, the Hemagglutination Inhibition (HI) test using AI and Newcastle Disease (ND) antisera yielded a negative result. Furthermore, the two-step RT-PCR on the passaged allantoic fluid, using H5 and H9 primers, was also negative. Based on the anatomical pathology observations, changes such as comb congestion, pectoral muscle hemorrhages, hepatic and cardiac necrosis, enteritis, and ovarian hemorrhages indicated that the chickens were infected with AI, even though the AI virus was not isolated or identified.

Kata Kunci : Avian Influenza, ayam layer, RT-PCR, hemaglutinasi (HA), hemaglutinasi-inhibisi (HI)/Avian Influenza, layer chicken, RT-PCR, hemagglutination (HA), hemagglutination inhibition (HI)