STUDI Indigofera tinctoria SEBAGAI ANTHELMINTIK PADA DOMBA YANG DIINFEKSI DENGAN Haemonchus contortus

ISKANDARMUDA, Dr. drh. Joko Prastowo, M.Si; Dr. drh. R. Wisnu Nurcahyo; Dr. drh. Sarmin, M.P

2021 | Disertasi | DOKTOR SAINS VETERINER

Haemonchosis mengakibatkan kerugian ekonomi yang besar terhadap peternak domba. Metabolit sekunder dari Indigofera tinctoria dapat digunakan sebagai anthelmintik. Penelitian ini bertujuan mengetahui angka prevalensi Haemonchus contortus pada domba, mengetahui pengaruh infusa I. tinctoria sebagai anthelmintik terhadap H. contortus domba secara in vitro dan secara in vivo. Penentuan prevalensi, sampel abomasum (200) diambil secara acak dari beberapa tempat pemotongan hewan di Sleman. Penentuan hewan positif didasarkan pada ditemukannya cacing dewasa H. contortus di abomasum. Uji in vitro, pada uji hambatan penetasan telur, suspensi telur (1000) di tempatkan dalam sumuran plat mikrotiter yang berisi 1 ml infusa I. tinctoria dosis 100, 120, 140, 160, 180, 200 dan 220 mg/ml, albendaole 1% dan Nacl fisiologis 0.62% dengan masing-masing 3 ulangan. Uji motilitas cacing dewasa, 10 cacing dewasa aktif di tempatkan dalam cawan petri yang berisi 25 ml infusa I. tinctoria dengan dosis 100, 120, 140, 160, 180, 200 dan 220 mg/ml, albendaole 1% dan Nacl fisiologis 0.62% dengan masing-masing 3 ulangan. Uji ultrastruktur permukaan tubuh cacing mati dengan scanning electron microscopy (SEM) dan profil protein dengan sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Uji in vivo, sebanyak 15 ekor domba di bagi rata menjadi 5 kelompok, semua kelompok diinfeksi 1000 L3 per oral setiap minggu selama 6 minggu. Pada minggu ke 9, kelompok 1 tidak diberi obat. Kelompok 2,3, dan 4 diberikan infusa I. tinctoria dengan dosis masing-masing 21 mg/ml, 41 mg/ml dan 62 mg/ml selama 9 hari. Kelompok 5 diberi albendazole 100 mg/ml. Pemeriksaan klinis, hematologi dan EPG dilakukan setiap minggu. Domba di autopsi dan dianalisa patologi. Uji SEM dilakukan pada cacing yang ditemukan pada abomasum. Prevalensi pada domba (68%). Data kuantitatif dianalisis statistik, hasil SEM, SDS-PAGE dan histopatolgi dianalisa deskriptif. Hasil in vitro, dosis infusa I. tinctoria 220 mg/ml menghambat penetasan paling tinggi (86%). Dosis infusa I. tinctoria 220 mg/ml membunuh cacing dewasa 93.33% setelah 8 jam perendaman. Hasil elektroforesis menunjukkan band protein pada dosis 22% lebih sedikit daripada kontrol. Hasil in vivo, rata-rata berat badan, BCS, FAMACHA, Hb, PCV dan RBC mengalami kenaikan. Infusa I. tinctoria menurunkan nilai FEC. Hasil SEM menunjukkan bahwa infusa I. tinctoria menyebabkan bagian anterior cacing menjadi mengkerut dengan lipatan yang tebal dan abrasi kutikula (in vitro), dan bagian posterior menyusut dengan adanya agregat pada kutikula (in vivo). Penelitian ini menunjukkan bahwa prevalensi H. contortus pada domba sebesar 68%, infusa I. tinctoria menghambat penetasan telur, menurunkan nilai FEC, meningkatkan berat badan, BCS, FAMACHA, Hb, PCV dan RBC, dan menurunkan profil protein cacing dewasa H. contortus. Kata Kunci: anthelmintik, Haemonchus contortus, Indigofera tinctoria, domba

Hemonchosis has resulted in huge economic losses for sheep farmers. Secondary metabolite compounds from Indigofera tinctoria L. can be used as anthelmintics. This study aimed to determine the prevalence of Haemonchus contortus on sheep and to evaluate the in vitro and in vivo effects of I. tinctoria L. aqueous extract (IAE) as an anthelmintic against Haemonchus contortus in sheep. Prevalence, abomasum samples (200) were collected randomly from the some local slaughterhouse in Sleman Regency, Yogyakarta. Determination of positif animals is based on the discovery of adult worms H. contortus in abomasum. In vitro test, in the egg hatch test, eggs suspension (1000 eggs) were placed in mikrotiter plat containing 1 ml of IAE at concentrations 100, 120, 140, 160, 180, 200, and 220 mg/mL, albendazole 1%, and 0.62% saline water each experiment was repeated. In the adult worm motility test, ten active adult worms were placed in each Petri dish containing 25 mL of IAE, each having a different concentration of IAE at concentrations 100, 120, 140, 160, 180, 200, and 220 mg/mL albendazole 1%, and 0.62% saline water each experiment was repeated. The ultrastructure of the egg was observed using dengan Scanning Electrone Microscopy (SEM) and protein profiles using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In the in vivo study, 15 sheep were used and divided into 5 groups. All groups were infected by 1000 infective larvaes per oral every week for 6 weeks. At 9 weeks after infection, group 1 untreated as a control. Groups 2,3,4 were given infusion doses 21 mg/ml, 41 mg/ml dan 62 mg/ml which given per day for 9 days. Group 5 was given albendazole. Clinical examination, hematology test and FECR were conducted every weeks. Sheeps were euthanized for post mortem examination. SEM was performed using worms found from the abomasum of slaughtered sheep. The prevalence of H. contortus was observed in sheep (68%). All quantitative data were analyzed statistically, data from SEM, SDS PAGE and histopathology were used for descriptive analysis. The results of in vitro, the results of egg hatch test, the 20% dose has greatest inhibitory hatchability (86%). The most effective concentration that killed the adult H. contortus worms was 220 mg/mL (93.33% mortality) after 8 h of treatment. The electrophoresis results showed that the protein band at a dose of 22% was less than that of the control. The results of in vivo, the average body weight, mean BCS, FAMACHA, Hb, PCV, TEC of post-treatment were increased. The aqueous extract of I. tinctoria was decresed FEC. The SEM results showed that IAE treatment caused the worms� anterior parts to become wrinkled with thick creases and cuticle abrasion (in vitro) and the anterior part to shrink along with the presence of aggregates in the worm cuticle (in vivo). This study showed that the prevalence H. contortus in sheep is 68%. The aqueous extract of I. tinctoria has an anthelmintic effect with inhibited hatchability, decreased FEC, increased weight gain, BCS, FAMACHA, Hb, PCV, and TEC, and reducing the protein profile of adult H. contortus worms. Keywords: anthelmintic, Haemonchus contortus, Indigofera tinctoria, scanning electron microscopy, sheep.

Kata Kunci : anthelmintik, Haemonchus contortus, Indigofera tinctoria, domba