Abstrak. Kaiin EM, Gunawan M, Octaviana S, Nuswantara S. 2017. Verifikasi molekuler metode sexing sperma sapi dengan kolom BSA (Bovine Serum Albumin). Pros Sem Nas Masy Biodiv Indon 3: [241][242][243][244][245]. Penentuan jenis kelamin anak sapi merupakan salah satu langkah strategis dalam perkembangan teknologi inseminasi buatan. Metoda pemisahan spermatozoa pembawa kromosom jenis kelamin betina (X) atau jantan (Y) dengan kolom BSA telah menghasilkan keberhasilan kesesuaian jenis kelamin anak sapi yang di lapangan sebesar 76% sampai 89%. Tujuan dari penelitian ini ialah memverifikasi hasil pemisahan sperma pembawa kromosom X dan kromosom Y dengan metode kolom BSA (Bovine Serum Albumine) secara molekuler. Sperma sexing sapi Simmental yang telah dipisahkan kromosom X dan Y dengan kolom BSA 5% (sperma X) dan 10% (sperma Y), diuji kualitas sperma secara mikroskopis meliputi parameter motilitas, viabilitas dan abnormalitas sperma. DNA sperma pada masing-masing kolom (5% atau 10%) diekstrasi menggunakan metoda spin-column dan kemudian diamplifikasi menggunakan Polymerase Chain Reaction (PCR) dengan design primer gen Sex-determining Region Y (SRY) yang berada pada daerah kromosom Y dan gen autosomal GADPH (Glyceraldehyde 3-phosphate dehydrogenase) pada daerah HMG Box (High Mobility Group Box). Hasil penelitian menunjukkan bahwa motilitas sperma X dan Y sebesar 60-70%, viabilitas sperma X sebesar 76,9-80,1% dan sperma Y sebesar 75,5-77,7%, sedangkan abnormalitas sperma X sebesar 6,4-7,6% dan sperma Y sebesar 5,2-5,5%. Hasil pemisahan sperma pada kolom BSA konsentrasi 10% dan sperma yang tidak disexing (kontrol), terverifikasi adanya 2 pita yaitu gen SRY (318 bp) dan GAPDH (415 bp). Hal ini menunjukkan bahwa pada kolom BSA 10% terdapat lebih banyak sperma Y. Hasil pada kolom BSA 5%, hanya terdapat 1 pita GAPDH (415 bp), menunjukkan sperma pada kolom tersebut adalah sperma X. Hasil ini menunjukkan bahwa sexing sperma sapi dengan metoda kolom BSA 5% dan 10%, dapat terverifikasi secara molekuler memisahkan sperma sapi pembawa kromosom X dan Y yang diuji dengan menggunakan metode duplex PCR.Kata kunci: Sexing, sperma, BSA, PCR, SRY Abstract. Kaiin EM, Gunawan M, Octaviana S, Nuswantara S. 2017. Molecular verification of sperm sexing method with BSA (Bovine Serum Albumin) column. Pros Sem Nas Masy Biodiv Indon 3: 241-245.Determining the sex of cattle was a strategic step in the development of artificial insemination technology. Separation of sperm carriers female sex chromosome (X) or male sex chromosome (Y ) methods with a BSA column has produced successful sex-matched calves after Artificial Insemination (AI) by 76% to 89%. The aim of this study was to verify the results of sperm separation with a BSA column method based on molecular technique. Simmental sperm was separated with 5% BSA column (X sperm) and 10% BSA column (Y sperm). Sperm quality was tested microscopically with parameters: motility, viability, sperm abnormalities and intact membrane plasma (IMP). DNA of sperm from each column was extracted using spin-column...

Four novel strains were isolated: PWU4T and PWU20T were both from soil in Germany, PWU5T was isolated from soil in India and PWU37T was obtained from sheep faeces collected on the Island of Crete. Cells of each were observed to be Gram-negative, strictly aerobic, rod shaped, and to grow optimally between 28 and 34 °C, between pH 7.0 and 8.0 and without the addition of NaCl. The strains were found to be catalase and oxidase-negative and able to grow on most mono- and disaccharides, a few polysaccharides and organic acids. Their predominant menaquinone was identified as MK-7. Their major fatty acids were identified as C16:1ω7c (PWU4T and PWU20T) and C16:1ω5c (PWU5T and PWU37T). The DNA G + C contents of strains PWU4T, PWU20T, PWU5T and PWU37T were determined to be 50.2 mol%, 51.6 mol %, 39.8 mol% and 53.8 mol%, respectively. The 16S rRNA gene sequence analysis revealed that the close relatives Ohtaekwangia koreensis 3B-2T and Ohtaekwangia kribbensis 10AOT share less than 93.8% sequence similarity. The strains were classified in two groups, where PWU4T and PWU20T share 93.0% sequence similarity, and PWU5T and PWU37T share 97.5% sequence similarity. However, the members of each group were concluded to represent different species based on the low average nucleotide identity (ANI) of their genomes, 69.7% and 83.8%, respectively. We propose that the four strains represent four novel species of two new genera in the family Cytophagaceae. The type species of the novel genus Chryseosolibacter is Chryseosolibacter histidini gen. nov., sp. nov. with the type strain PWU4T (= DSM 111594T = NCCB 100798T), whilst strain PWU20T (= DSM 111597T = NCCB 100800T) is the type strain of a second species, Chryseosolibacter indicus sp. nov. The type species of the novel genus Dawidia is Dawidia cretensis gen. nov., sp. nov. with the type strain PWU5T (= DSM 111596T = NCCB 100799T), whilst strain PWU37T (= DSM 111595T = NCCB 100801T) is the type stain of a second species, Dawidia soli sp. nov.

Mangroves are unique intertidal ecosystems that provide ecological niches to different microbes, which play various roles in nutrient recycling and diverse environmental activities. The association between myxobacteria and mangroves are hitherto poorly understood. The aim of our study was to evaluate the myxobacterial community composition as well as isolate myxobacteria and to characterize the antimicrobial activity of myxobacteria isolates from Indonesian mangroves. Twenty-five cultivable myxobacteria were affiliated in six genera: Myxococcus, Corallococcus, Archangium, Chondromyces, Racemicystis and Nannocystis of the order Myxococcales based on partial 16S rRNA gene sequences. Thirteen crude extracts showed moderate activities against at least one of human pathogenic microorganisms. The crude extract of Racemicystis sp. strain 503MSO indicated a novel compound, which has not been reported in the database yet and the identification of this compound needs further study. The myxobacterial communities of three different sampling sites were analyzed using primers adapted for the myxobacteria group identification. The results showed that myxobacterial communities are more diverse than assumed. Therefore, our study has highlighted the importance of the mangrove habitat as promising harbor of myxobacteria as well as novel antimicrobial compounds with activity against pathogenic microorganisms.

The area within Pusbindiklat Peneliti-LIPI (The National Training and Education Center for Researchers Development-Indonesian Institute of Sciences) has been planted with many trees as an effort for reforestration and increasing aesthetic value. Unfortunately, the management of Pusbindiklat Peneliti-LIPI paid less attention to the trees potency and its characteristic as well as inventorizing all trees. This study aim to inventorize tree species based on their morphological characteristic in order to know the potency concerned with safety. Research was conducted through inventorizing and scoring the suitability factors of all trees, determining the tree plantation points, which is converted to the map of Pusbindiklat Peneliti-LIPI. In addition, direct interviews were also conducted to the civitas of Pusbindiklat Peneliti-LIPI. Trees that have been found in the area of Pusbindiklat Peneliti-LIPI consist of 20 family, 42 species, and 217 individuals. From the inventory of the trees, there were 11 species which were unsuitable with the criteria of urban forest vegetation parameter. Moreover, based on the map of trees species at Pusbindiklat Peneliti-LIPI area, there are 85 individual from 20 species of trees that are not suitable to be planted in Pusbindiklat Peneliti, since they were planted near the parapet wall and buildings of Pusbindiklat Peneliti-LIPI.

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