Siti Chotiah
Balai Besar Penelitian Veteriner, Bogor

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Pengaruh Proses Freeze-Drying dan Penyimpanan pada Suhu Kamar terhadap Viabilitas dan Patogenisitas Plasma Nutfah Mikroba Pasteurella Multocida Siti Chotiah
Buletin Plasma Nutfah Vol 12, No 1 (2006)
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/blpn.v12n1.2006.p40-44

Abstract

The effect of freeze-drying process and preserving in a vacuum at room temperature against viability and pathogenicity of veterinary microbe germ plasma of Pasteuerella multocida BCC 2331 was investigated at Balitvet. The aim of this study was to find out the most effective and efficient conservation method. As much as 5,2 x 1011 colony forming unit (CFU)/ml of bacteria suspension in 7.5% glucose serum as the preservation medium being pathogenic in mice with LD50 of 9,8 CFU/ml was freeze dried then stored at room temperature (±27oC) until the study was completed. Viability and pathogenicity test were done immediately after the process, 1 and 2 months after storage. The results showed that there were viability decreases amounted 1,3 x 101 CFU/ml, 102 CFU/ml and 8,2 x l02 CFU/m1 due to the effects of the process, one month and two-month storage respectively. The decreases of pathogenicity on mice were shown by the increases of LD50 amounting log 1, log 2, and log 3 a day after the process, one month and two-month storage respectively. AbstrakPengaruh proses kering beku dan penyimpanan hasil proses pada suhu kamar 27oC terhadap viabilitas dan patogenisitas plasma nutfah mikroba veteriner telah dipelajari di Balitvet untuk menentukan cara pelestarian yang efektif dan efisien. Dalam kegiatan ini dipakai bakteri Pasteurella multocida koleksi Balitvet Culture Collection nomor koleksi B2331. Suspensi bakteri sebanyak 5,2 x l011 coloni forming unit (CFU)/ml dalam medium preservan 7,5% glukosa, serum dan bersifat patogen pada mencit dengan LD50 9,8 CFU/ml diproses kering beku, kemudian disimpan pada suhu kamar (+27oC) sampai penelitian selesai. Uji viabilitas dan patogenisitas dilakukan langsung setelah proses dan pada 1 serta 2 bulan setelah penyimpanan. Hasil penelitian menunjukkan terjadi penurunan viabilitas sebanyak 1,3 x 101 CFU, dan 8,2 x 102 CFU/ml masingmasing karena pengaruh proses, pengaruh penyimpanan selama 1 dan 2 bulan. Patogenisitas pada mencit menurun yang ditandai oleh adanya peningkatan LD50 sebanyak log 1, log 2, dan log 3 masing-masing 1 hari setelah proses, 1 dan 2 bulan setelah penyimpanan.
Pemanfaatan Plasma Nutfah Mikroba Bordetella bronchiseptica sebagai Perangkat Deteksi Antibodi Siti Chotiah
Buletin Plasma Nutfah Vol 14, No 2 (2008): December
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/blpn.v14n2.2008.p90-96

Abstract

Microbe germplasm of Bordetella bronchiseptica has been found in pigs in Indonesia and recognized as the causal agent of atrophic rhinitis and pneumonia, and also one of the agents involved in porcine respiratory disease complex. Those microbes have been characterized and conserved ex situ at Balitvet Culture Collection. The aim of this research was to find out seroepidemiological of B. bronchiseptica infections at four pig herds in two regencies in Central Java, by utilizing the germ plasma as antibody detection kit. The ELISA technique used lypopolysaccharide and sera hypperimmune of B. bronchiseptica local isolate as an antigen and positive sera respectively. Referring to cut off level of 0.404, the survey results showed that of 25.4% of 63 pig serum samples examine by ELISA were positively infected by B. bronchiseptica; 14.29% and 11, 11% from Karanganyar regencies and Sragen regencies respectively. Base on pig ages it showed that 10.35%, 24%, and 77.8% to be seropositive from less than three-month age group, three-month until five-month age group, and more than five-month age group respectively. Base on pig farm it was showed that 28.60; 30; 13.64; and 40% were seropositive collected from Farm 1, Farm 2, Farm 3 and Farm 4 respectively. The results indicated that B. bronchiseptica infection was spread on four pig herds in two regencies in Central Java with the higest seropositive (77,8%) in more than five-month age group. AbstrakPlasma nutfah Bordetella bronchiseptica telah ditemukan pada babi di Indonesia, selain merupakan agen penyebab atrophic rhinitis dan pneumoni juga salah satu agen porcine respiratory disease complex. Mikroba tersebut telah dikarakterisasi dan dikonservasi ex situ di Balitvet Culture Collection (BCC). Tujuan penelitian adalah untuk mengetahui seroepidemiologi infeksi B. bronchiseptica pada empat peternakan babi di dua kabupaten di Jawa Tengah, dengan memanfaatkan plasma nutfah tersebut sebagai bahan baku perangkat deteksi antibodi. Teknik Enzyme-linked Immunosorbent Assay (ELISA) menggunakan lypopolysaccharide dan serum imun dari isolat lokal B. bronchiseptica BS(BCC 2455) masing-masing sebagai antigen dan serum kontrol positif. Mengacu pada nilai cut off 0,404, hasil survei menunjukkan bahwa 25,4% dari 63 sampel serum yang diperiksa positif terinfeksi oleh B. bronchiseptica, 14,3% di antaranya berasal dari Kabupaten Karanganyar dan 11,1% dari Kabupaten Sragen. Berdasarkan pengelompokan umur diketahui bahwa seropositif sebanyak 10,35; 24; dan 77,8% masing-masing berasal dari kelompok umur di bawah 3 bulan, kelompok umur 3 sampai 5 bulan, dan kelompok umur di atas 5 bulan. Berdasarkan peternak sebaran seropositif sebanyak 28,60; 30; 13,64; dan 40% masing-masing pada peternakan 1, 2, 3, dan 4. Hal ini menunjukkan infeksi B. bronchiseptica telah tersebar di empat peternakan babi di dua kabupaten di Jawa Tengah dengan seropositif tertinggi (77,8%) pada kelompok babi umur di atas 5 bulan.