Judul: Genomic DNA analysis of the Heat Shock Protein 70 as a  molecular marker candidate of Temperature-Shock Resilience in Osphronemus  goramy
Abstrak: Temperature stress often causes high mortality in giant gourami. One strategy to prevent this threat is to use giant gourami that are adaptive to temperature stress. Adaptive fish can be produced by molecular marker-based selection that utilizes genes associated with giant gourami resilience traits. However, the availability of gene sequence information and molecular markers for selection has not been widely done. This research was conducted as an initial stage for the development of molecular marker-based selection of gourami adaptive to temperature stress through gene expression analysis and gene identification. This research was carried out in three stages. The first stage aims to analyze gene expression levels related to gourami resilience. The second stage aims to identify markers in the gourami genome related to resilience. The third stage aims to select gourami using the Tetra primer amplification refractory mutation system-polymerase chain (Tetra-ARM PCR) method and evaluate its correlation with fish resilience. The study began with raising 200 fish to be conditioned in culture containers. For gene expression analysis, 20 giant gourami were transferred to a 55 cm × 45 cm × 45 cm aquarium with a water level of 35 cm. The fish were kept for fourteen days at 28 - 30°C and fed commercial feed with a protein content of 39% by mouth. Water changes of 30% of the aquarium volume were done once a day. The temperature stress test was conducted in two aquarium containers measuring 55 cm × 45 cm × 45 cm with a water height of 35 cm. Each aquarium had its water temperature regulated with the first aquarium at a hot temperature of 32-34 °C and the second aquarium at a cold temperature of 18-20 °C. The fish were transferred directly from the rearing container to the first aquarium for one hour, then the fish were transferred to the second aquarium for one hour. Furthermore, fish behavior was observed and differentiated into two groups of fish, namely adaptive fish and susceptible fish. Adaptive fish are fish that can still swim well, while vulnerable fish are fish that have collapsed at the bottom of the aquarium. A total of five adaptive fish and five vulnerable fish were taken for total RNA extraction. Expression analysis was performed on three resilience-related genes, namely heat shock protein 70 (HSP70), Cu-superoxide dismutase (CuSOD), dan Mn-superoxide dismutase (MnSOD) using the qPCR method. The results showed that adaptive fish had a higher gene expression level than the susceptible fish for all the genes tested. The results indicate that there is an important role of these genes in the response of giant gourami to temperature stress. Therefore, these genes can be used as a potential gene as a molecular marker for giant gourami selection. The second phase of the study aims to identify single nucleotide polymorphisms (SNPs) in the HSP70 genome sequence and evaluate fish survival after temperature stress testing. The preparation of adaptive and susceptible fish giant gourami samples for HSP70 genome sequence analysis was carried out in stages, with three replicates. A total of 40 giant gourami were used at each stage. Temperature
Keyword: genotype, HSP70, molecular marker, Osphronemus goramy, SNPs

Judul: Gene Expression Analysis of Heat Shock Protein-70 in Giant Gourami after Transportation Stress
Abstrak: Respons fisiologis pada tubuh ikan yang terpapar stresor salah satunya adalah sintesis heat shock protein-70 (HSP70). Sintesis HSP70 merupakan respons dari sel yang berfungsi menjaga kelengkapan protein sel, pemindahan protein, menjaga dari pelipatan prematur pada sel dan mencegah penumpukan protein pada sel. Penelitian ini bertujuan untuk menganalisis tingkat ekspresi gen HSP70 pada benih ikan gurami yang terpapar stres transportasi. Penelitian dilakukan dengan mentransportasikan ikan perlakuan (3 ulangan) dengan plastik kemas berukuran 40 × 60 cm2 berisi ikan dengan panjang 6,02±0,62 cm dan bobot 8,28±0,49 g dengan kepadatan 40 ikan per-plastik. Ikan kontrol (1 ulangan) dikemas dengan spesifikasi yang sama seperti ikan perlakuan dan diletakkan dalam akuarium tanpa ditransportasikan. Ikan kemudian langsung dibedah pascatransportasi dan diambil sampel hati (liver) untuk ekstraksi RNA dan kemudian disintesis menjadi cDNA. Sampel cDNA lalu diamplifikasi dengan metode PCR menggunakan primer HSP70 dan β-aktin. Tingkat ekspresi gen ditentukan menggunakan metode PCR semikuantitatif. Hasil elektroforesis menunjukkan bahwa ukuran pita pada sampel primer HSP70 sebesar 267 bp dan primer β-aktin sebesar 192 bp. Analisis ekspresi gen HSP70 pada ikan pasca paparan stres transportasi menunjukkan bahwa tidak ada perbedaan nyata antara ikan kontrol, rentan dan tahan stres setelah dilakukan uji statistik (P>0,05). Stres transportasi tidak memberikan pengaruh yang signifikan terhadap ekspresi gen HSP70 pada hati ikan gurami., One of the physiological responses of fish exposed to stress transportation is the synthesis of heat shock protein-70 (HSP70). Synthesis of HSP70 is a cell response to stressors that function to maintain the completeness of cell proteins, transfer proteins, prevent premature folding of cells, and prevent protein accumulation in cells. This research aimed to analyze the expression level of the HSP70 gene in giant gourami fry exposed to transportation stress. The research was conducted by transporting treated fish (3 replicates) with plastic containers measuring 40 × 60 cm2 containing fish with a length of 6.02±0.62 cm and a weight of 8.28±0.49 g and a density of 40 fish per plastic. Control fish (1 replicate) were packed with the same specifications as the treatment fish and placed in the aquarium without being transported. The fish were then dissected after transport and liver samples were taken for RNA extraction and then synthesized into cDNA. The cDNA samples were then amplified by the PCR method using HSP70 and β-actin primers. Gene expression level was determined by using the semi-quantitative PCR method. Electrophoresis results showed that the band size in the HSP70 primer sample was 267 bp and the β-actin primer was 192 bp. HSP70 gene expression analysis in giant gourami after being exposed to transportation stress showed that there was no significant difference between control, prone to-, and stress-resistant fish after statistical tests (P>0.05). Transport stress did not have a significant effect on the expression of the HSP70 gene in giant gourami liver.
Keyword: Giant Gourami, HSP70, Primer, Stress, Transportation

Judul: Peranan Pemimpin Informal dalam Menggerakan Partisipasi Masyarakat  untuk Pembangunan Desa
Abstrak: Pendekatan pembangunan yang banyak diterima para ahli adalah pendekatan dari bawah ke atas (bottom up) yang berarti bahwa dalam pembangunan diperlukan keikutsertaan rakyat. Keikutsertaan tidak hanya pada tahap merencanakan pembangunan, akan tetapi juga ikut dalam melaksanakan, bahkan sampai pada menilai hasil-hasil pembangunan (Cohen dan Uphoe 1977; Dusseldorp, 1981; Slamet, 1993). Agar rakyat yang berhimpun dalam kelompok-kelompok dapat ikut serta, mereka harus memiliki sejumlah kualifikasi yang diperlukan untuk itu.
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