Ita Djuwita

mtDNA Variation and Human-Mediated Introgression of Indigenous Sus Populations on Several Indonesian Islands

Abstract. To examine the genetic origin of the domestic pig, the distribution of wild boar, and human-mediated translocation of the domestic pig, we collected 223 samples from domestic pigs and wild boars from eight Indonesian islands, sequenced the control region of mitochondrial DNA (mtDNA) from each sample, and compared these sequences with previously determined sequences from East and Southeast Asian domestic pigs and wild boars. Three Sus species (S. scrofa, S. verrucosus, and S. celebensis) were identified on the Indonesian islands. The mtDNA sequences of three Indonesian Sus species were diverse, and they clustered into three lineages with low bootstrap values (an S. scrofa group including East and Southeast Asian domestic pigs and wild boars, a group including indigenous S. scrofa together with S. verrucosus from Sumatra and Java Islands, and an S. celebensis group from Sulawesi Island). The mtDNA haplotypes of S. scrofa wild boars from three (Sumbawa, Flores and New Guinea) islands and domestic pigs from two (Lombok and Timor) islands east of the Wallace Line, and some S. scrofa wild boars from Sumatra and Java Islands were related to Vietnamese pig mtDNA sequences in the East and Southeast Asian domestic pig and wild boar clade, supporting that ancient immigrants likely introduced domestic pigs from the Asian continent to east Indonesian islands. The mtDNA haplotypes of S. celebensis were broadly divided into three groups, which were distributed in the north and southwest areas, central area and southeast area of Sulawesi Island.

In Vitro Fertilization and Embryo Development of Vitrified Ovine Oocytes Stressed in Sucrose

Experiments were conducted on the morphology, fertilization and embryo development rate of vitrified ovine oocytes matured in vitro. Three vitrification solutions were used for vitrification. PBS supplemented with 1% BSA, 30% ethylene glycol was added by one of three different sucrose concentrations, 1.00 M (VS 1 ), 0.50 M (VS 2 ), and 0.25 M (VS 3 ). The results showed that the percentages of normal vitrified oocytes after warming were 78 and 63% in VS 1 and VS 2 , respectively, which was significantly higher as compared for VS 3 . The fertilization rates were 59 and 66% in VS 1 and VS 2 , respectively, which were also significantly higher as compared with VS 3 (35%). Zygote viability after 18 h was 57; 43; and 40%, for VS 1 ,VS 2 , and VS 3 , respectively, which was not significantly different. The incidence of polyspermic penetration increased with increasing sucrose concentration, i.e 23, 11, and 9% in VS 1 , VS 2 , and VS 3 , respectively, as compared with unvitrified oocytes (4%). The cleavage rate of vitrified oocytes in VS 1 was 13.2% which was significantly lower (p<0.05) compared to those of unvitrified control oocytes (70.0%). Hence, a high sucrose concentration is beneficial for maintaining the oocyte structure during the processes of vitrification and thawing, which ultimately results in increased in vitro fertilization rates.

RESPONS SEL EPITEL USUS (CMT-93) TERHADAP NUTRASETIKAL GALOHGOR

Penelitian ini bertujuan membandingkan pengaruh nutrasetikal galohgor dalam bentuk serbuk (GS) dan ekstrak (GE) terhadap proliferasi, morfologi, dan ekspresi gen Aldh1a2 pada sel epitel usus (CMT-93). Galohgor serbuk (GS) dibuat dengan menghancurkan semua bahan dan dikeringkan menggunakan drum-dryer sedangkan GE dibuat dengan mengeringkan semua bahan dengan oven, digiling, dan dimaserasi dengan etanol selama 3x24 jam. Perlakuan didasarkan pada konsentrasi akhir β-karoten yang berasal dari GS dan GE masing-masing sebesar 0,5; 1,5; dan 5,0 µM dalam larutan medium Dulbeccos Modified Eagles medium (DMEM) yang dilengkapi serum (10%). Analisis proliferasi menggunakan asai 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dan ekspresi gen dianalisis dengan reverse transcriptase polymerase chain reaction (RT-PCR). Hasil penelitian meunjukkan bahwa GE pada konsentrasi tinggi (5,0 μM) secara signifikan (P<0,05) dapat menekan proliferasi dan memengaruhi morfologi sel CMT-93. Beta-karoten dalam GE dan GS memengaruhi ekspresi gen Aldh1a2 pada sel epitel usus CMT-93

In Vitro Fertilization and Embryo Development

Sperm collected from cauda epididymis is a source of male gametes. The purposes of this study was to evaluate an quality of frozen-thawed sperm of garut ram which was collected from cauda epididymis and cryopreserved with modified Tris extender, i.e: Tris extender (control, KT), Tris extender + 60 mM lactose (LS), and Tris extender + 60 mM lactose + 0.05% glutathione (GL). Quality of collected sperm including concentration, motility, live sperm, abnormality, cytoplasmic droplet, intact acrosomal cap (IAC), and intact plasma membrane (IPM) were evaluated. Results showed that mean of sperm concentration, percentages of motility, live sperm, abnormality, cytoplasmic droplet, IAC, and IPM of fresh epididymal sperm were 13,993.33 million/ml, 70.83, 82.83, 10.83, 8.5, 85.83, and 81.33%, respectively. Sperm quality after equilibration for LS and GL were significantly (PExperiments were conducted on the morphology, fertilization and embryo development rate of vitrified ovine oocytes matured in vitro. Three vitrification solutions were used for vitrification. PBS supplemented with 1% BSA, 30% ethylene glycol was added by one of three different sucrose concentrations, 1.00 M (VS1), 0.50 M (VS2), and 0.25 M (VS3). The results showed that the percentages of normal vitrified oocytes after warming were 78 and 63% in VS1 and VS2, respectively, which was significantly higher as compared for VS3. The fertilization rates were 59 and 66% in VS1 and VS2, respectively, which were also significantly higher as compared with VS3 (35%). Zygote viability after 18 h was 57; 43; and 40%, for VS1,VS2, and VS3, respectively, which was not significantly different. The incidence of polyspermic penetration increased with increasing sucrose concentration, i.e 23, 11, and 9% in VS1, VS2, and VS3, respectively, as compared with unvitrified oocytes (4%). The cleavage rate of vitrified oocytes in VS1 was 13.2% which was significantly lower (p