Kumalaningrum, Dwi Retna and Dr.Uun Yanuhar, S.Pi.,M.Si and Prof.Dr.Ir Muhammad Musa, MS (2021) Respon Imunologis Indigenous Sitokin (Interleukin-6, Interleukin-10 dan INOS) Pada Ikan Kerapu Cantang (Epinhepelus sp.) Yang Diinfeksi Viral Nervous Necrosis (VNN) Dengan Treatmen Brachionus sp. Doctor thesis, Universitas Brawijaya.
Abstract
Ikan kerapu cantang merupakan komuditas andalan untuk dibudidayakan karena memiliki harga jual yang tinggi. Namun tingginya harga jual ini menyebabkan eksploitasi secara berlebihan dan peningkatan budidaya. Upaya peningkatan budidaya tidak berarti tanpa adanya masalah yang menyebab penurunan produksi. Salah satu masalah yang dihadapi yaitu serangan penyakit yang disebabkan oleh virus. Virus yang sering menginfeksi ikan kerapu yaitu Viral Nervous Necrosis. Serangan virus ini menyebabkan kematian masal pada berbagai tahapan perkembangan spesies ikan air laut terutama pada stadia larva dan benih, bahkan mampu menyebabkan kematian hingga 100 %. Upaya yang dilakukan untuk meminimalisir serangan VNN ini dengan pemberian immunomodulator. Salah satu kandidat yaitu Brachionus sp. Brachionus sp. memiliki kandungan lemak kasar sebesar 17,17 %, protein kasar sebesar 63,53 %, karbohidrat 7.74 %, phospholipid 44 %, EPA 41 gr/kg, DHA 66 gr/kg serta asam amino yang berlimpah meliputi asam glutamate (Glu) 62 gr/kg berat kering; lisina (Lys) 41,2 gr/kg berat kering; asam aspartat (Asp) 39,2 gr/kg berat kering; dan Leusina (Leu) 38,8 gr/kg berat kering. Tingginya protein yang terkandung pada brachionus dapat dijadikan sebagai kandidat imunomodulator. Hal ini dikarena protein dapat memacu pengerahan sel inflamasi sehingga meningkatkan respon sistem kekebalan tubuh ikan yang tahan terhadap penyakit. Hal ini disebabkan bila terjadi malnutrisi akibat kekurangan protein mampu menyebabkan penurunan imun dalam fagositosis. Tujuan dari penelitian ini yaitu untuk mengetahui respon indigenous sitokin IL-6, IL-10 dan iNOS pada ikan kerapu di infeksi virus Viral Nervous Necrosis dengan treatmen Brachionus sp. Penelitian ini dilaksanakan pada bulan Januari- September 2021 di Laboratorium Biokimia dan patologi Fakultas Kedokteran Universitas Brawijaya, Laboratorium Biosains Universitas Brawijaya, Laboratorium Reproduksi ikan fakultas perikanan dan ilmu kelautan Universitas Brawijaya, laboratorium Anatomi Universitas Airlangga PSDKU Banyuwangi dan CV. SAA Banyuwangi. Metode yang digunakan dalam penelitian ini adalah eksperimental. Tahapan yang dilakuakn untuk pelaksaan penelitian meliputi kultur Brachionus sp, ekstraksi protein Brachionus sp. dan identifikasi protein Brachionus sp. kemudian dilakuakn pemeliharaan ikan secara invivo, penyuntikan protein Brachionus sp. dan pengambilan organ target (otak). setelah dilakukan pengambilan organ target (otak), maka dilakukan analisa flowcytometri IL-10 dan IL-6 dan Imunohistokimia IL-10, IL-6 dan INOS. Selama pemeliharaan in vivo, juga dilakukan pengamatan kualitas air. Hasil dari penelitian adalah hasil kuantifikasi protein dari Brachionus sp. didapatkan hasil kandungan protein supernatant sebesar 92,9 mg/ml dengan nilai absorbansi 1.67 dan kuantifikasi protein untuk pellet Brachionus sp. sebesar 61.5 mg/ml dengan nilai absorbansi 1.12. Hasil identifikasi protein dengan SDS-page didapatkan 3 pita dengan berat molekul sebesar 13,77kDa, 75,49 kDa dan 122,73kDa dengan indikasi jenis protein yang terkandung yaitu peridinin, apyrase dan threonine. Pada dosis pemberian protein Brachionus sp. 35μl, 105μl,107μl menunjukkan bahwa protein yang terkandung pada Brachionus dapat menurunkan sitokin pro-inflamasi IL-6 dan INOS dan anti-inflamasi IL-10. Hasil kualitas air selama pengamatan termasuk optimal untuk kehidupan ikan kerapu pada setiap perlakuan yaitu suhu rata-rata 29,17±0,16 0C, pH rata-rata sebesar 7,69±0,16, DO rata-rata sebesar 5, 36±0,08 dan salinitas rata-rata sebesar 33,53±0,09. Penelitian ini dapat disimpulkan bahwa pemberian protein pada ikan kerapu cantang mampu menunjang pertumbuhan ikan kerapu cantang yang meliputi pertambahan panjang dan berat tubuh ikan serta pembentukan imun. Selain itu, pemberian protein Brachionus mampu menurunkan kadar ekspresi IL-6, IL-10 dan INOS, dimana penurunan ekspresi ini menandakan bahwa proses inflamasi didalam tubuh mengalami penurunan. Oleh karena itu, saran untuk penelitian selanjutnya dengan melakukan purifikasi protein Brachionus sp dengan diuji ditantang virus VNN pada ikan kerapu cantang untuk melihat pengaruhnya terhadap sitokin interleukin-6, interleukin-10 dan INOS. Selain itu, perlu dilakukan penelitian lanjutan tentang dosis yang mampu menyebabkan autoimun pada ikan kerapu yang ditreatment dengan protein Brachionus sp.
English Abstract
Cantang grouper is a superior commodity to be cultivated because it has a high selling price. However, this high selling price has led to overexploitation and increased cultivation. Efforts to increase cultivation are meaningless without problems that cause production declines. One of the problems faced is the attack of diseases caused by viruses. The virus that often infects groupers is Viral Nervous Necrosis. This virus attack causes mass death at various stages of development of marine fish species, especially in the larval and seed stages, even being able to cause death up to 100%. Efforts are being made to minimize this VNN attack by administering an immunomodulator. One of the candidates is Brachionus sp. Brachionus sp. contains crude fat of 17.17%, crude protein of 63.53%, carbohydrates 7.74%, phospholipids 44%, EPA 41 g/kg, DHA 66 g/kg and abundant amino acids including glutamic acid (Glu) 62 g /kg dry weight; lysine (Lys) 41.2 g/kg dry weight; aspartic acid (Asp) 39.2 g/kg dry weight; and Leusina (Leu) 38.8 g/kg dry weight. The high protein contained in brachionus can be used as an immunomodulator candidate. This is because the protein can stimulate the mobilization of inflammatory cells thereby increasing the response of the fish's immune system that is resistant to disease. This is because if there is malnutrition due to protein deficiency, it can cause a decrease in immunity in phagocytosis. The purpose of this study was to determine the response of indigenous cytokines IL-6, IL-10 and iNOS in grouper fish to Viral Nervous Necrosis virus infection with Brachionus sp. This research was conducted in January-September 2021 at the Laboratory of Biochemistry and Pathology, Faculty of Medicine, Universitas Brawijaya, Laboratory of Biosciences, Universitas Brawijaya, Laboratory of Fish Reproduction, Faculty of Fisheries and Marine Sciences, Universitas Brawijaya, Laboratory of Anatomy Universitas Airlangga PSDKU Banyuwangi and CV. SAA Banyuwangi. The method used in this research is experimental. The steps for carrying out the research include Brachionus sp culture, Brachionus sp protein extraction. and identification of Brachionus sp. then rearing the fish in vivo, by injecting Brachionus sp. and target organ (brain) harvesting. After the target organ (brain) was removed, a flowcytometry analysis of IL-10 and IL-6 and Immunohistochemistry of IL-10, IL-6 and INOS was performed. During in vivo maintenance, water quality observations were also carried out. The results of the study were the results of protein quantification from Brachionus sp. The results showed that the supernatant protein content was 92.9 mg/ml with an absorbance value of 1.67 and protein quantification for pellets of Brachionus sp. of 61.5 mg/ml with an absorbance value of 1.12. The results of protein identification with SDS-page obtained 3 bands with molecular weights of 13.77 kDa, 75.49 kDa and 122.73 kDa with an indication of the type of protein contained, namely peridinin, apyrase and threonine. At the dose of protein Brachionus sp. 35μl, 105μl,107μl showed that the protein contained in Brachionus can reduce pro-inflammatory cytokines IL-6 and INOS and anti- inflammatory IL-10. The results of water quality during the observation were optimal for grouper life in each treatment, namely the average temperature of 29.17±0.16 0C, the average pH of 7.69±0.16, the average DO of 5.36± 0.08 and the average salinity was 33.53±0.09. This research can be concluded that the provision of protein in cantang grouper can support the growth of cantang grouper which includes the increase in length and weight of the fish as well as the formation of immunity. In addition, the administration of Brachionus protein was able to reduce the expression levels of IL-6, IL-10 and INOS, where this decrease in expression indicated that the inflammatory process in the body had decreased. Therefore, the suggestion for further research is to purify Brachionus sp protein by being challenged with the VNN virus in cantang grouper to see its effect on the cytokines interleukin-6, interleukin-10 and INOS. In addition, further research is needed on doses capable of causing autoimmune in groupers treated with Brachionus sp protein.
| Item Type: | Thesis (Doctor) |
|---|---|
| Identification Number: | 0621080008 |
| Subjects: | 600 Technology (Applied sciences) > 639 Hunting, fishing & conservation > 639.3 Culture of cold-blooded vertebrates |
| Divisions: | S2/S3 > Magister Budidaya Perairan, Fakultas Perikanan dan Ilmu Kelautan |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 11 Jan 2022 07:45 |
| Last Modified: | 24 Feb 2022 03:46 |
| URI: | http://repository.ub.ac.id/id/eprint/188172 |
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