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      • KCI등재

        Development of RAPD-PCR assay for identifying Holstein, Angus, and Taiwan Yellow Cattle for meat adulteration detection

        Chin-Cheng Lin,Pin-Chi Tang,Hsin-I Chiang 한국식품과학회 2019 Food Science and Biotechnology Vol.28 No.6

        Incidents of food fraud have occurred worldwide,particularly in the form of meat adulteration. In thisstudy, molecular probes were developed using the Randomamplification of polymorphic DNA (RAPD) polymerasechain reaction (PCR) technique in order to identify threebeef subspecies–Holstein, Angus, and Taiwan YellowCattle. Four RAPD-PCR 10-nucleotide primers were chosenout of a total of 60 primers. The selection was based onthe reproducibility of species-specific amplicons able todetect various origins of cattle breeds. The resultsdemonstrated that primer OPK12 produced three uniqueamplicons (1100 bp, 1000 bp and 480 bp) in Holstein;primer OPK14 generated one amplicon that only appearedin Holstein and Angus (200 bp); primer OPK19 amplifiedtwo species-specific amplicons in Holstein measuring550 bp and 650 bp, respectively. However, due to therelatively lower repeatability of RAPD-PCR, higher andmore specific testing repeats were required to increase theaccuracy of the conclusion.

      • SCIESCOPUSKCI등재

        Biochemical Compositions of Follicular Fluid and the Effects of Culture Conditions on the In Vitro Development of Pig Oocytes

        Huang, Wei-Tung,Lu, She-Ghi,Tang, Pin-Chi,Wu, Shinn-Chih,Cheng, San-Pao,Ju, Jyh-Cherng Asian Australasian Association of Animal Productio 2002 Animal Bioscience Vol.15 No.10

        The aims of this study were, firstly, to analyze the biochemical compositions of serum and follicular fluid (FF) from prepubertal gilts after PMSG (1,000 IU) treatment. The concentrations of total proteins, lipids, cholesterol, glucose and sex hormones (progesterone, $P_4$; estradiol-$17{\beta}$, $E_2$; testosterone, T) were measured. Secondary, the effects of porcine FF (pFF) addition (40% and 100%) in IVM media and different culture conditions [Exp. 1: mBMOC-2+20% porcine serum (PS), fresh IVM medium, filtered IVMconditioned medium, or rabbit oviducts; Exp. 2: mBMOC-2+20%PS or stepwise medium replacement procedures (SMRP) cocultured with or without cumulus cells] on the in vitro development (IVD) of porcine oocytes were also examined. Results showed that no significant differences were found in total protein levels between serum and pFF from different sizes (large, >7 mm; medium, ~5-7 mm; small, <3-5 mm) of follicles (75-85 and 49-90 mg/dl; p>0.05). Total lipid concentrations remained constant in serum (395-472 mg/dl), and reduced significantly in the pFF from large follicles (287 mg/dl) at 132 h after PMSG treatment when compared to those at other time points (441-480 mg/dl). Basal cholesterol levels in serum and pFF at 12 h were similar (153-161 mg/dl), but increased at 36 h (186-197 mg/dl). Basal P4 and E2 levels in serum (0.1 ng/ml and 5.5 pg/ml) were low, but increased from 0.34 ng/ml and 12.13 pg/ml at 24 h to 0.81 ng/ml and 61.70 pg/ml at 98 h, respectively, after PMSG treatment (p<0.05). P4 levels increased linearly in pFF from large follicles during 12 through 132 h (138-1,288 ng/ml). A similar increase was also observed in $E_2$ levels (22-730 pg/ml) before 60 h post PMSG treatment, and then dropped afterwards (730-121 pg/ml). The development of the oocytes fertilized in 40% pFF-medium was greater than that in 100% pFF-medium group without gonaodtropin addition (31% vs 10%, p<0.05). However, both were lower than those in mBMOC-2+20%PS and in rabbit oviducts (p<0.05). When cocultured with cumulus cell monolayers, a greater cleavage rate was observed in the group cultured in filtered IVM-conditioned medium than the SMRP group (36% vs 18%, p<0.05). A similar phenomenon was also observed in the culture without cumulus cell monolayers (33% vs 19%, p<0.05). It is concluded that neither the fresh IVM nor filtered IVM-conditioned medium has positive effect on the IVD of oocytes. Coculture with cumulus cell monolayers and the SMRP were not beneficial to the development of IVF pig oocytes.

      • SCIESCOPUSKCI등재

        The Relative Centrifugation Force Permits Visualization of the Germinal Vesicle in Pig Oocytes

        Hsieh, Chang-Hsing,Lee, Stone,Jaw, Si-Ning,Tseng, Jung-Kai,Tang, Pin-Chi,Chang, Lan-Hwa,Ju, Jyh-Cherng Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.9

        Pig oocytes contain high levels of lipids in the ooplasm, which reduces the visibility of the germinal vesicle (GV) under microscopic examination. Therefore, the purposes of this study were to investigate the effects of relative centrifugation force (RCF) on the visibility and maturation rates of the GV stage oocytes after centrifugation. In Experiment 1, cumulus-oocyte-complexes (COCs) were collected from slaughterhouse ovaries and randomly allocated to different RCFs (3,000 rpm: 970 g; 6,000 rpm: 3,900 g; or 10,000 rpm: 10,840 g) for 10 or 20 min. Percentages of visible GV were 76-79% in the oocytes centrifuged with 10,000 rpm, which were significantly higher (p<0.01) than those with 3,000 and 6,000 rpm. No significant differences in GV visibility were observed among oocytes with different lengths of centrifugation (p<0.05) regardless of the RCFs. In esperiment 2, the maturation rate of the oocyte was found significantly lower in the 20 min than in the 10 min group received 10,840 g of RCF (30 vs. 75%, p<0.05). In conclusion, the GV of porcine oocytes can be clearly visible by centrifugation at 10,840 g for 10 min without compromising their subsequent maturation rates and a longer centrifugation time (20 min) had no beneficial influence on the visibility of GV stage pig oocytes.

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