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THE APPLICATION OF UTY AND SRY MOLECULAR MARKERS FOR DETERMINATION OF UNKNOWN SEX SAMPLES IN BALI CATTLE Indriawati, Indriawati; Volkandari, Slamet Diah; Margawati, Endang Tri
Jurnal ILMU DASAR Vol 21 No 1 (2020)
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (86.781 KB) | DOI: 10.19184/jid.v21i1.9333

Abstract

An investigation involving large number of animals is often resulting incomplete or in accurate information such as animal parentage, or misidentify on sex due to unlabeled sex samples. A PCR method by applying Y chromosome markers (UTY and SRY) facilitates in determination of unknown sex problem. This study was intended to determine sex from unlabelled sex of blood samples by applying PCR method using a pooled-DNA template. Twenty five of unknown sex blood samples from Nusa Penida, Bali were used in this study. The samples were plotted into 5 pooled-DNA whith each pool DNA consisted of 5 individuals DNA. Two pairs of sex primers, UTY (58oC) and SRY (60oC) with 35 cycles were applied to amplify the samples. The result showed there was only one pooled-DNA (P4) amplified by UTY (484bp). Whereas re-PCR of the positive pooled-DNA (P4) using SRY primer, only one out of 25 samples determined as male Bali cattle (325bp). This finding suggests that UTY and SRY primers are suitable for sex determination and the pooled-DNA could be used as an efficient PCR method both in consumables and PCR process for sex determination. Keywords: Determination, sex, unknown sample, pooled DNA, Bali cattle.
Genetic Diversity and Relationship among Bali Cattle from Several Locations in Indonesia Based on ETH10 Microsatellite Marker Margawati, Endang Tri; Volkandari, Slamet Diah; Indriawati, Indriawati; Ridwan, Muhamad
Jurnal Ilmu Ternak dan Veteriner Vol 23, No 4 (2018): DECEMBER 2018
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (172.619 KB) | DOI: 10.14334/jitv.v23i4.1915

Abstract

Bali cattle is one of local beef cattle in Indonesia, up to present its performance indicated an inbreeding occurrence. This study was aimed to analyze the genetic diversity and relationship among Bali cattle from several locations in Indonesia based on ETH10 microsatellite marker. Ninety-four (94) DNA samples (89 Bali cattle; 5 Banteng) were analyzed. The Bali cattle samples were from 6 locations in Indonesia (15 Pulukan; 15 Nusa Penida; 14 Bima West Nusa Tenggara/WNT; 10 Mataram, WNT; 20 Riau; 15 South Borneo). DNA Banteng samples were collected from Prigen Malang of East Java. Microsatellite marker of ETH10 labelled HEX was used for amplification. Alleles were analyzed by using Cervus 3.0.7 and GenAlex 6.5. Result showed that there were five (5) alleles found in ETH10 marker i.e., 209; 213; 215; 217; and 219 bp. Average of observed (Ho) and expected (He) heterozygosity were 0.46±0.05 and 0.60±0.03, respectively. Five (5) out of 6 locations were in breeding occurrence except Bali cattle from Mataram was not inbreeding. The longest genetic relationship was between Bali cattle from Mataram and Riau whereas the closest distance was Bali cattle from South Borneo with Mataram. Banteng was closest to Bali cattle from Nusa Penida and the longest was to Bali cattle from South Borneo. This finding indicates there is inbreeding in Bali cattle, therefore it needs to be concerned in bull rotation and semen distribution for increasing the Bali cattle performance.
Effect of maturation periods and leukaemia inhibitory factor on in vitro bovine embryo development Margawati, Endang Tri
Indonesian Journal of Animal and Veterinary Sciences Vol 1, No 3 (1995)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (612.166 KB) | DOI: 10.14334/jitv.v1i3.26

Abstract

The period of in vitro maturation,~20 vs 24 hours) with of without supplementation of :,~uk_ni;ia inhib toryy factor (LIF) (0, 500, 1000 or 2000 U/ml) was studied on bovine embryo development in vitro in a 2 x 4 factotial experiment w id oestgnoo i, a randomized block design . A total of 870 bovine oocytes were used . Besides embryo development, cell numbers of blastocysts v. : ;re also co , mted in order to study the quality of the embryos . Oocytes were matured in a modified TCM199 medium containing 10 ug/nil of FS-14 and LP., I 1~ t1L rstradiol, fertilized in TALP and cultured in SOF/AABSA medium. There was no interaction between maturation periods and LIF doses on embryo development (P>0 .05). Maturation periods, however, affected (P<0.05) blastocyst rates but did not for cleavage o, oocytes and the percentage of oocytes that developed into blastocysts . LIF doses during in vitro maturation did not affect embryo development (P>0 .05) . Cell numbers of blastocysts were also not affected by maturation periods and LIF doses (P<0 .05), however 20 h in vitro maturation and supplementation of LIF doses tended to increase the cell numbers. This study suggests that 20 h maturation increases blastocyst rates and that supplementation with LIF during maturation does not affect the quality of embryos produced in vitro. Key words : in vitro fertilization, oocyte, cleavage, blastocyst, cell numbers of blastocyst
GENETIC VARIATION OF CALPASTATION GENE OF INDIGENOUS BALI CATTLE (BOS JAVANICUS) IN INDONESIA Margawati, Endang Tri; Volkandari, Slamet Diah; Indriawati, Indriawati; Svensson, Emma M.
HAYATI Journal of Biosciences Vol. 26 No. 1 (2019): January 2019
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (508.086 KB) | DOI: 10.4308/hjb.26.1.44

Abstract

Calpastatin is one of gene markers affecting meat tenderness. The study aimed to evaluate genetic variation of calpastatin (CAST) gene of Bali cattle (Bos javanicus) in lndonesia. A total of 61 samples consisting of 21 Bali cattle, 22 Ongole cattle (Bos indicus), and 18 Friesian Holstein (FH) cattle (Bos taurus) were applied. The Ongole and FH cattle were involved for breed comparison. DNA was extracted from fresh blood using a High Salt method and measured their quality by a Spectrophotometer. A 523 bp of Calpastatin gene fragment was amplified by Polymerase Chain Reaction and Restriction Fragment Polymorphism (PCR-RFLP) technique with RsaI restriction enzyme for genotyping. Result showed that two variants alleles (C and G) and three genotypes (CC, GC, GG) were found in those Bali, Ongole and FH samples. Allele G was dominant allele with the highest G allele was in Bali cattle population (0.88). The higher percentage of allele C was found in Ongole and Friesian Holstein compared to that in Bali cattle. The Ongole breed tends to have a potential source of lean meat quality. This finding identified that genetic variation of CAST gene was exist in Bali cattle and adapted cattle of Ongole and FH in Indonesian.
PENGUJIAN PENCEMARAN DAGING BABI PADA BEBERAPA PRODUK BAKSO DENGAN TEKNOLOGI PCR: PENCARIAN SISTEM PENGUJIAN EFEKTIF Margawati, Endang Tri; Ridwan, Muhamad
BERITA BIOLOGI Vol 10, No 1 (2010)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v10i1.2055

Abstract

Entering globalization market, Indonesian government could not reject any import of food products from overseas. To anticipate the possibility of porcine contaminants into processed meat products of imported food such as meat or chicken ball, sausage, tin meat etc., it is important to apply laboratory research on such particular matter in regard to ethical and certain religious concern. This study was intended to identify the possibility of porcine contaminants into either processed meat products or fresh meat.A technique of polymerase chain reaction (PCR) was applied and PCR optimizing was conducted in advanced to obtain the right annealing temperature.Positive control of fresh pork meat was amplified to get porcine Leptin size which is 152bp fragment. Five samples of 4 meat balls and one fresh beef meat were individually collected for their DNA by either from minced or mashed after liquid nitrogen exposure then followed with a series of DNA extraction steps. PCR was assigned by using a specific primer of Leptin gene for porcine identification.Visualization of Leptin fragment was applied either on 1%, 2% of agarose gel or 10-20% gradient acrylamide gel.The result showed that all sample applied were not identified for containing porcine contaminants while positive control was on the right size of 152bp of Leptin gene. Specific primer used in this study was proved that there was not identified porcine Leptin gene on the negative control (fresh beef meat). This study suggests that a method of PCR is a simple analytical method for identification of porcine contaminants and visualization on 2% agarose gel is a cheaper and quicker method while by gradient acrylamide gel showing more clear band however this method is time consuming and expensive.
IN VITRO DEVELOPMENT OF OVINE EMBRYOS FOLLOWING MATURATION UNDER LIMITED CO2 MARGAWATI, ENDANG TRI
HAYATI Journal of Biosciences Vol. 12 No. 3 (2005): September 2005
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (58.347 KB) | DOI: 10.4308/hjb.12.3.112

Abstract

An experiment was conducted to examine the influence of CO2 during in vitro oocyte maturation on the in vitro ovine embryo development. Three treatments of CO2 were subjected to the oocyte development. Those were 2h gasses prior to maturation in incubator (T1); without CO2 either prior to or over maturation (T2) and CO2 exposure both prior to and over 22h maturation (T3). A total of 324 oocytes were used. Putative zygotes were cultured for seven days and evaluated for their developmental stage. Presence of CO2 (T3) increased the proportion of oocytes reaching Metaphase II ( 66.50 + 3.5%; p
Effect of maturation periods and leukaemia inhibitory factor on in vitro bovine embryo development Margawati, Endang Tri
Jurnal Ilmu Ternak dan Veteriner Vol 1, No 3 (1995)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (612.166 KB) | DOI: 10.14334/jitv.v1i3.26

Abstract

The period of in vitro maturation,~20 vs 24 hours) with of without supplementation of :,~uk_ni;ia inhib toryy factor (LIF) (0, 500, 1000 or 2000 U/ml) was studied on bovine embryo development in vitro in a 2 x 4 factotial experiment w id oestgnoo i, a randomized block design . A total of 870 bovine oocytes were used . Besides embryo development, cell numbers of blastocysts v. : ;re also co , mted in order to study the quality of the embryos . Oocytes were matured in a modified TCM199 medium containing 10 ug/nil of FS-14 and LP., I 1~ t1L rstradiol, fertilized in TALP and cultured in SOF/AABSA medium. There was no interaction between maturation periods and LIF doses on embryo development (P>0 .05). Maturation periods, however, affected (P<0.05) blastocyst rates but did not for cleavage o, oocytes and the percentage of oocytes that developed into blastocysts . LIF doses during in vitro maturation did not affect embryo development (P>0 .05) . Cell numbers of blastocysts were also not affected by maturation periods and LIF doses (P<0 .05), however 20 h in vitro maturation and supplementation of LIF doses tended to increase the cell numbers. This study suggests that 20 h maturation increases blastocyst rates and that supplementation with LIF during maturation does not affect the quality of embryos produced in vitro.
KOLEKSI DNA DAN KONFIRMASI MARKA ETH10 PENGKODE SIFAT PERTUMBUHAN PADA SAPI PASUNDAN Ningsih, Heni Utami; Praakarsa, Tatag Bagus Putra; Margawati, Endang Tri
BIOTROPIC The Journal of Tropical Biology Vol 1 No 1 (2017): Biotropic, Volume 1, Nomor 1, 2017
Publisher : Program Studi Biologi, Fakultas Sains dan Teknologi, Universitas Islam Negeri Sunan Ampel Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Sapi Pasundan secara genetik merupakan generasi dari tetua yang mengalami tekanan inbreeding dari program grading up sapi Ongole dan program grading up sapi Jawa dengan sapi Madura dan sapi Bali. Sifat pertumbuhan merupakan salah satu sifat penting bernilai ekonomis. Salah satu kandidat marka genetik pengkode sifat pertumbuhan yaitu marka mikrosatelit ETH10. Tujuan dari penelitian ini yaitu memahami prosedur konfirmasi marka ETH10 pengkode sifat pertumbuhan pada sapi Pasundan. Sebanyak tiga (3) sampel darah sapi Pasundan (619, 626, dan 602) diambil dari vena caudalis yang digunakan dalam isolasi DNA dengan metode garam pekat. DNA yang dikoleksi kemudian dikuantifikasi menggunakan spektrofotometer untuk mengetahui konsentrasi DNA (λ260) dan kemurnian DNA (λ260/λ280). Amplifikasi marka ETH10 menggunakan metode Polymerase Chain Reaction (PCR) dengan suhu annealing 60oC dan 35 siklus. Produk PCR gen (fragment) ETH10 divisualisasi pada Gel Agarose 1,5%. Hasil isolasi DNA diperoleh dengan rata-rata konsentrasi DNA sebesar 116,37± 63,37 ng/μl dan kemurnian DNA sebesar 1,72±0,035. Selanjutnya hasil Amplifikasi marka DNA ETH10 pada sapi Pasundan menunjukkan ukuran fragment sebesar 212-224 pb pada ke tiga sampel. Kesimpulan dari penelitian ini yaitu telah terkonfirmasinya marka ETH10 yang berhubungan dengan sifat pertumbuhan pada sampel sapi Pasundan. Hasil tersebut dapat digunakan sebagai informasi awal dalam analisis genetik sapi Pasundan
IDENTIFIKASI VIRUS PENYAKIT JEMBRANA PADA SAPI BALI MENGGUNAKAN PENANDA MOLEKULER GEN env SU [Identification of Jembrana Disease Virus by Using a Molecular Marker of env SU Gene in Bali Cattle] Indriawati, Indriawati; Margawati, Endang Tri; Ridwan, Muhammad
BERITA BIOLOGI Vol 12, No 2 (2013)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (182.055 KB) | DOI: 10.14203/beritabiologi.v12i2.534

Abstract

Up to present, detection of Jembrana disease virus has been identified through serological test. Advances in molecular biology has enabled to detect Jembrana disease virus earlier, quicker and more accurate by application of molecular markers.The aim of this study was to identify Jembrana disease by using molecular marker of env SU gene in Bali cattle.Total RNA of Jembrana disease virus (7732bp) was collected from spleen of Bali cattle suspected Jembrana disease by using RNEasy Protect Mini Kit (QIAGEN). A pair of specific primers was designed from Jembrana viral genome (env SU) that accessed through a GenBank with Accession Number of U21603.A kit of Access Quick RT-PCR System (PROMEGA) was used for Reverse-Transcriptase-PCR (RT-PCR). The RT-PCR products were visualized on 2% agarose gel.The result showed a single band with the size of ± 900bp in all samples. This size indicated that env SU gene was existed in the examined spleen samples. This finding suggests that a molecular marker could be used accurately to identify the env SU gene in JDV of Bali cattle.
GENETIC POLYMORPHISM OF CALPASTATIN (CAST) GENE IN PASUNDAN CATTLE Volkandari, Slamet Diah; Nadila, Aina; Radiastuti, Nani; Margawati, Endang Tri
Buletin Peternakan Vol 42, No 4 (2018): BULETIN PETERNAKAN VOL. 42 (4) NOVEMBER 2018
Publisher : Faculty of Animal Science, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (450.456 KB) | DOI: 10.21059/buletinpeternak.v42i4.35338

Abstract

The aim of this study was to determine genetic diversity of Calpastatin gene (CAST) in Pasundan cattle. Forty-four fresh blood samples were collected from UPTD BPPT Beef Cattle Ciamis West Java. Genomic DNA samples were extracted using High Salt method. A 523 bp fragment of Calpastatin gene was successfully amplified using PCR method. Genotyping of CAST gene was conducted by PCR-RFLP method using RsaI restriction enzyme (GT^AC). Genotypes and alleles were analyzed using software Cervus 3.0.7. Parameters were observed i.e genotypes and alleles frequencies, heterozygosity observed (Ho) and expected (He), Hardy Weinberg Equilibrium (HWE), and Polymorphic Information Content (PIC). Result showed that three variant genotypes of GG, GC and CC were found and two variant alleles of G and C. Allele G was found higher (0.77) than allele C (0.23). Population of Pasundan cattle was found polymorphism and in the Hardy Weinberg Equilibrium. Polymorphic Information Content (PIC) value showed in a moderate (0.290) condition. Values of Heterozygosity observed and expected were 0.409 and 0.355 respectively. This research concludes that there is polymorphism of CAST gene in Pasundan cattle population and has genetic diversity. This result could be used as early genetic information in exploration of Pasundan cattle.