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An improvement of real-time polymerase chain reaction system based on probe modification is required for accurate detection of African swine fever virus in clinical samples in Vietnam

  • Tran, Ha Thi Thanh;Dang, Anh Kieu;Ly, Duc Viet;Vu, Hao Thi;Hoang, Tuan Van;Nguyen, Chinh Thi;Chu, Nhu Thi;Nguyen, Vinh The;Nguyen, Huyen Thi;Truong, Anh Duc;Pham, Ngoc Thi;Dang, Hoang Vu
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.10
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    • pp.1683-1690
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    • 2020
  • Objective: The rapid and reliable detection of the African swine fever virus (ASFV) plays an important role in emergency control and preventive measures of ASF. Some methods have been recommended by FAO/OIE to detect ASFV in clinical samples, including realtime polymerase chain reaction (PCR). However, mismatches in primer and probe binding regions may cause a false-negative result. Here, a slight modification in probe sequence has been conducted to improve the qualification of real-time PCR based on World Organization for Animal Health (OIE) protocol for accurate detection of ASFV in field samples in Vietnam. Methods: Seven positive confirmed samples (four samples have no mismatch, and three samples contained one mutation in probe binding sites) were used to establish novel real-time PCR with slightly modified probe (Y = C or T) in comparison with original probe recommended by OIE. Results: Both real-time PCRs using the OIE-recommended probe and novel modified probe can detect ASFV in clinical samples without mismatch in probe binding site. A high correlation of cycle quantification (Cq) values was observed in which Cq values obtained from both probes arranged from 22 to 25, suggesting that modified probe sequence does not impede the qualification of real-time PCR to detect ASFV in clinical samples. However, the samples with one mutation in probe binding sites were ASFV negative with OIE recommended probe but positive with our modified probe (Cq value ranked between 33.12-35.78). Conclusion: We demonstrated for the first time that a mismatch in probe binding regions caused a false negative result by OIE recommended real-time PCR, and a slightly modified probe is required to enhance the sensitivity and obtain an ASF accurate diagnosis in field samples in Vietnam.

UV-B Effects on Growth and Nitrate Dynamics in Antarctic Marine Diatoms Chaetoceros neogracile and Stellarima microtrias (중파 자외선에 노출된 남극 규조 Chaetoceros neogracile와 Stellarima microtrias의 성장과 질산염 흡수량의 변화)

  • Gang, Jae Sin;Gang, Seong Ho;Lee, Yun Ho;Sim, Jeong Hui;Lee, Sang Hun
    • ALGAE
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    • v.18 no.1
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    • pp.13-20
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    • 2003
  • Two isolated Antarctic marine diatoms, Chaetoceros neogracile VanLandingham and Stellarima microtrias (Ehrenberg) Hasle and Sims were examined to show changes of growth and uptake rate of nitrate due to UV-B irradiance. Chlorophyll (chl) a concentration was regarded as the growth index of diatom. The diatoms were treated with UV-B radiation and cultured for 4 days under cool-white fluorescent light without UV-B radiation. Two levels of UV-B exposures were applies: 1 and 6 W $m^{-2}$. Durations of UV-B treatment were 20, 40 and 60 minutes under 6 W $m^{-2}$ and 1, 2, 3, 4 and 5 hrs under 1 W $m^{-2}$. The control groups were cultured at the same time without UV-B radiation. The growth rates of two diatoms decreased under 1 and 6 W $m^{-2}$ UV-B irradiances than that of control group. After 4 days, chl a concentrations of C. neogracile were increased more than 4 times from 133 μgo$l^{-1}$ to 632 μgo$l^{-1}$ in control group. However, the concentration of experimental groups under 1 W $m^{-2}$ UV-B were only increased from 139 μgo$l^{-1}$ to 421 μgo$l^{-1}$ during one hour and the chl a concentrations were decreased from 144 μgo$l^{-1}$ to 108 μgo$l^{-1}$ during five hour. Growth of diatom dramatically more decreased under 6 W $m^{-2}$ UV-B than 1 W $m^{-2}$ UV-B. The chl a concentration of experimental groups under 6 W $m^{-2}$ UV-B for one hour was only increased from 111 μgo$l^{-1}$ to 122 μgo$l^{-1}$. In the case of S. microtrias showed also similar pattern to C. neogracile by UV-B radiation. The uptake rates of nitrate by the two strains were decreased abruptly under 6 W $m^{-2}$ UV-B irradiances. When two strains were treated under 1 and 6 W $m^{-2}$ UV-B during one hour, the strains were only continued growth and uptake of nitrate under 1 W $m^{-2}$ UV-B. This experimental evidence shows that exposure to UV-B radiation especially to high irradiance of UV-B decreases diatom survival and causes lower decrease of nutrient concentrations by microalgae in Antarctic water. Furthermore, evidence suggests that microalgal communities confined to near-surface waters in Antarctica will be harmed by increased UV-B radiation, thereby altering the dynamics of Antarctic marine ecosystems.

A Study on the Childbirth Experience in Primiparas (출산경험에 대한 연구)

  • Kim, Hyun-Kyung
    • Women's Health Nursing
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    • v.1 no.1
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    • pp.81-96
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    • 1995
  • This study was intended to provide women who are supposed to experience childbirth with effective nursing care exploring the childbirth experience. The purpose of this paper is to give an answer to the question of what the childbirth experience is, and phenomenological method has been used for that purpose. 17 primiparous mothers right after delivery in one university hospital of J city were served as subjects for this research. Data were collected June to October, 1993 through in depth interview with subjects using unstructured and open questions about the childbirth experience. Van Kaam's phenomenological analysis method was used for the analysis of the data. The results of the study are summarized as follows : The contents of the experience which primiparous mothers had undergone through childbirth were pain, fear, worry, relief, lightness, thankfulness, unsatisfactory, unreality, holiness for a new life, identifying the meaning of life, becoming a mother. 1. Experience before delivery Mothers experienced pain, fear, worry, unsatisfactory, relief, and thankfulness before delivery. Subjects' descriptions about the pain appeared to be various ; unbearable pain, bearable pain, anticipated pain. And their reactions were also various ours ; some endured pain, sought alternative method i.e., surgery, didn't want to reexperience it, or accepted it as women's fate. Subjects experienced fear for anticipated pain, suffering pain, and possible delayed delivery progress and were worried about delivery progress and baby wellbeing. Also, Subjects were unsatisfied with professionals' unfaithful attitude, their spouses' absence, and the ignorance of their pain. But subjects became relieved at the accustomed surroundings, good progress of delivery, support of family, care of professionals and support of other family. And they expressed their thanks to the professionals and family members for doing their best and also to their mothers who gave birth to them. 2. Experience after delivery Subjects experienced lightness, thankfulness, unsatisfactory, worry, unreality, holiness for a new life, identifying the meaning of life, becoming a mother after delivery. Mothers experienced lightness from the fact that it is over, and freshness right after delivery. They were thankful that they delivered vaginally and had an easy delivery. Subjects who delivered female baby were unsatisfied and some were worried about rearing the baby. By confessing, "I have no specific feeling, I am very confused, It is unrealistic, I feel strange", they expressed the unrealistic aspect of childbirth experience. Subjects felt holiness for a new life for novelty and birth of life. They identified the meaning of life from the fact that they felt worthy, fulfilled their duty, had their own baby, accomplished an important affair. After they experienced what it is to be a mother, they realized with it is being a mother and had a rearing expectation for the baby. The results of the study will provide basic data for caring the childbearing women.

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Nitrogen Balance in Goats Fed Flemingia (Flemingia Macrophylla) and Jackfruit (Artocarpus Heterophyllus) Foliage Based Diets and Effect of a Daily Supplementation of Polyethylene Glycol (PEG) on Intake and Digestion

  • Mui, Nguyen Thi;Ledin, Inger;Uden, Peter;Binh, Dinh Van
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.5
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    • pp.699-707
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    • 2002
  • Diets with foliage of Flemingia (Flemingia macrophylla) or Jackfruit (Artocapus heterophyllus were fed to goats with the objective to study nitrogen (N) balance and effect of a daily supplementation of polyethylene glycol (PEG) on intake and digestion. In experiment 1, three male Alpine${\times}$Jamnapary goats with initial weights varying from 26.9 to 27.7 kg were used in a $3{\times}3$ Latin square design in the dry season. Three Alpine${\times}$Bachthao crosses, 15.3-16.7 kg, were used in the same design in the wet season. The three diets were based on chopped whole sugar cane complemented with the two green foliages, Jackfruit and Flemingia, or soybean meal (SBM). The level of dry matter (DM) offered was 4% of body weight (BW), 2.7% as foliage and 1.3% as chopped whole sugar cane. The amount of SBM offered was calculated to give the same amount of crude protein (CP) as the foliages. Each experimental period lasted 32 days (14 days for adaptation, 7 days for collection and 10 days for rest). Feed intake, apparent digestibility of DM, organic matter (OM), CP, neutral detergent fiber (NDF) and acid detergent fiber (ADF) and retained nitrogen (N) were measured by total faecal and urine collection. In experiment 2, four male goats (Alpine${\times}$Jamnapary) with initial weights from 17.1 to 23.1 kg were used in a $4{\times}4$ Latin square design. The four treatments were Jackfruit or Flemingia with or without addition of PEG, which was fed at a level of 5 g/goat and day by mixing with a small amount of rice bran. Each experimental period lasted 15 days (8 days for adaptation, 7 days for collection). Measurements were done as in experiment 1. The DM digestibility was highest (65.9-74.3%) for goats fed the SBM diet in both the dry and wet season. The DM digestibility of goats fed the Jackfruit and the Flemingia diets was similar in both the dry (58.6-59.2% respectively) and the wet season (53.9-56.1% respectively). The CP digestibility was highest (73.0-73.6%) for the SBM diet followed by the Jackfruit diet (47.0-38.5%) and was lowest (36.8-30.0%) for the Flemingia diet in both dry and wet seasons, respectively. The NDF digestibility was low for both the Jackfruit (36.4%) and Flemingia (38.0%) diets in the wet season. All diets resulted in a positive N balance. The N retention was highest (0.465-0.604 g/kg $W^{0.75}$) in the SBM diets and lowest (0.012-0.250 g/kg $W^{0.75}$) in the Flemingia diet. Addition of PEG had no effect on feed intake for any of the diets. PEG added in the Flemingia diet had a positive effect only on NDF digestibility, but the digestibility of the Jackfruit diet was significantly increased. Supplementation with PEG reduced digestibility and N retention of Flemingia, possibly because of the low tannin level, but increased digestibility and N retention for Jackfruit foliage.

Determination of Carnitine Renal Threshold and Effect of Medium-Chain Triglycerides on Carnitine Profiles in Newborn Pigs

  • Heo, K.N.;Odle, J.;Lin, X.;van Kempen, T.A.T.G.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.2
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    • pp.237-242
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    • 2001
  • Colostrum deprived, newborn pigs (N=12, $1.64{\pm}0.05kg$) were used to study the renal threshold of carnitine, and effects of emulsified medium-chain triglyceride (MCT, tri-8:0) feeding on kinetics of plasma carnitine and urinary carnitine excretion. An arterial catheter was inserted through an umbilical artery, and a bladder catheter was inserted via the urachus. Piglets were oro-gastrically gavaged with one of six carnitine levels (0, 60, 120, 180, 240, $480{\mu}mol/kg\;W^{0.75}$) with (+MCT) or without medium-chain triglycerides (-MCT) in 0.9% NaCl solution. Blood was sampled into heparinized tubes at 0, 1, 2, 4, 6, 8, 14, and 20 h after gavage, and urine was collected and pooled into 1 h or 2 h composite samples to determine free- and short-chain carnitine concentrations. Plasma from the 12 newborn piglets before gavage contained $10.6{\pm}1.2{\mu}mol/L$ free carnitine and $7.2{\pm}0.6{\mu}mol/L$ acid-soluble acyl carnitine. The renal threshold for carnitine was similar between the MCT and the +MCT group (42.6 13.1 and $46.4{\pm}2.0{\mu}mol/L$, respectively), but the correlation between plasma free carnitine and urinary excretion was altered. Plasma free carnitine linearly increased with increasing carnitine dosage (-MCT group, $R^2=0.95$, p<0.001; +MCT group, $R^2=0.91$, p<0.001), but was decreased by 50% when medium-chain triglycerides were fed. The peak in plasma free carnitine concentration was depressed by medium-chain triglycerides feeding also. Therefore, the plasma and urinary short-chain/free carnitine ratio of the +MCT group was increased by 100% and 40%, respectively (p<0.01). Feeding of medium-chain triglycerides may delay plasma carnitine elevation via altering the kinetics of absorption. Similarly, the plasma and urinary short-chain/free carnitine ratio were affected by interaction between medium-chain triglycerides and time (p<0.01). The present study suggests that an oral carnitine dose over $480{\mu}mol/kg\;W^{0.75}$ may be needed to reach the free carnitine renal threshold within a short period, especially when provided together with medium-chain triglyceride.

Genetic parameters for milk yield in imported Jersey and Jersey-Friesian cows using daily milk records in Sri Lanka

  • Samaraweera, Amali Malshani;Boerner, Vinzent;Cyril, Hewa Waduge;Werf, Julius van der;Hermesch, Susanne
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.11
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    • pp.1741-1754
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    • 2020
  • Objective: This study was conducted to estimate genetic parameters for milk yield traits using daily milk yield records from parlour data generated in an intensively managed commercial dairy farm with Jersey and Jersey-Friesian cows in Sri Lanka. Methods: Genetic parameters were estimated for first and second lactation predicted and realized 305-day milk yield using univariate animal models. Genetic parameters were also estimated for total milk yield for each 30-day intervals of the first lactation using univariate animal models and for daily milk yield using random regression models fitting second-order Legendre polynomials and assuming heterogeneous residual variances. Breeding values for predicted 305-day milk yield were estimated using an animal model. Results: For the first lactation, the heritability of predicted 305-day milk yield in Jersey cows (0.08±0.03) was higher than that of Jersey-Friesian cows (0.02±0.01). The second lactation heritability estimates were similar to that of first lactation. The repeatability of the daily milk records was 0.28±0.01 and the heritability ranged from 0.002±0.05 to 0.19±0.02 depending on day of milk. Pearson product-moment correlations between the bull estimated breeding values (EBVs) in Australia and bull EBVs in Sri Lanka for 305-day milk yield were 0.39 in Jersey cows and -0.35 in Jersey-Friesian cows. Conclusion: The heritabilities estimated for milk yield in Jersey and Jersey-Friesian cows in Sri Lanka were low, and were associated with low additive genetic variances for the traits. Sire differences in Australia were not expressed in the tropical low-country of Sri Lanka. Therefore, genetic progress achieved by importing genetic material from Australia can be expected to be slow. This emphasizes the need for a within-country evaluation of bulls to produce locally adapted dairy cows.

Health-Care Providers' and Parents' Perspectives on Complementary Alternative Medicine in Children with Cancer in Indonesia

  • Susilawati, Dwi;Sitaresmi, Mei;Handayani, Krisna;Ven, Peter van de;Sutaryo, Sutaryo;Kaspers, Gertjan;Mostert, Saskia
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3235-3242
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    • 2016
  • Background: Complementary alternative medicine (CAM) use in children with cancer is widespread. Health-care providers (HCP) need to acknowledge and address this need. This study explored and compared perspectives on CAM of HCP and parents of young patients with cancer in Indonesia. Materials and Methods: We conducted a cross-sectional study using semi-structured questionnaires in HCP and parents of childhood cancer patients at an Indonesian academic hospital. Results: A total of 351 respondents participated: 175 HCP (response rate 80%) and 176 parents (response rate 80%). Parents were more likely than HCP to think that chemotherapy can cure cancer (80% compared to 69%, P=0.013). Nearly half of all parents (46%) and HCP (45%) doubted whether CAM can cure cancer. Parents were more likely than HCP to think that CAM can be helpful in childhood cancer treatment (54% compared to 35%, P=0.003). The most recommended CAM by HCP was self-prayer (93%). Reasons for recommending CAM were: hope for improvement of the child's condition (48%), patient wants to stop treatment (42%). Most discouraged CAM by HCP was by old-smart people (70%), the reasons being: lack of evidence for usefulness (77%), lack of CAM knowledge (75%). The proportion thinking that patients were unlikely to raise the CAM topic if they perceived that doctors were skeptical was higher in parents than in HCP (52% versus 1%) (P<0.001). Most HCP (71%) and parents (77%) acknowledged that their knowledge about safety and efficacy of CAM was inadequate (P=ns). The proportion that wanted to learn or read more about CAM was higher among parents than HCP (48% compared to 31%, P=0.002). Conclusions: HCP and parents have different perspectives on CAM use in children with cancer. HCP should enhance their CAM knowledge and encourage open communication about CAM with parents. If doctors' skepticism is perceived, parents are unlikely to raise CAM as a topic.

The Influences of Addition of Sugar with or without L. buchneri on Fermentation and Aerobic Stability of Whole Crop Maize Silage Ensiled under Anaerobic Silos

  • Guan, Wu-tai;Driehuis, F.;van. Wikselaar, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.8
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    • pp.1128-1133
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    • 2002
  • The whole plant of crop maize was chopped and ensiled in airtight 1-L capacity glass jars to determine the influence of residual sugar on anaerobic yeast growth and on the fermentation of lactic acid by L. buchneri in whole crop maize silage. There were a total of six treatments used in this experiment as follow: added 25 g de-mineralised water per kg chopped maize serving as control (con), 37.5 g glucose solution containing 12.5 g glucose ($g_1$), 75 g glucose solution containing 25 g glucose ($g_2$), 25 g L. buchneri suspension intended for $10^6$ cfu $g^{-1}$ (L.b.), $g_1+L.b.$ and $g_2+L.b.$ All silos were stored in the dark at $20^{\circ}C$ until end of experiment. Jars were opened on duplicates at day 2, 7, 14, 28, 56 or triplicates at day 91 after ensiling for measuring the pH, microbiological enumeration and fermentative products. Results indicated that acidification rates for all silages were very fast, no difference occurred among treatments before day 28. After day 28 the pH values for silages inoculated by L. buchneri. with or without sugar tended to increase especially for treated only with L. buchneri, resulting in higher (p<0.01) finial pH than uninoculated silages. Compared with control silage, the added sugar significantly (p<0.01) increased dry matter (DM) loss, L. buchneri enhanced (p<0.01) DM loss further at different sugar existence. Silages inoculated by L. buchneri only or in combination with sugar addition contained less (p<0.01) lactic acid than the correspondent silages without inoculation with L. buchneri. In comparison with control, ethanol production is about 3 or 6 fold higher due to addition 12.5 or 25 g glucose per kg chopped maize at ensiling. The added sugar resulted in less acetic acid concentration (p<0.01) than control, but inoculation with L. buchneri increased (p<0.01) acetic acid than correspondent uninoculated silages at different sugar levels. No butyric acid and propionic acid were found in uninoculted silages, silages inoculated with L. buchneri. produced more propionic acid, 1-propanol and butyric acid. Lactobacilli counts were not influenced by added sugar, but increased (p<0.01) with inoculation of L. buchneri. The added sugar increased significantly (p<0.01) the yeast count, whereas L. buchneri showed the contrary effect. No differences were found in the aerobic stability among all treatments. In conclusions, 1) the added sugars encourage the growth of yeast and yeasts convert extra sugar into ethanol in maize silages. 2) The added sugars and L. buchneri do not influence the aerobic stability of silages stored in anaerobic silos.

Effect of Work and Urea-Molasses Cake Supplementation on Live Weight and Milk Yield of Murrah Buffalo Cows

  • Van Thu, Nguyen;Uden, Peter
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.9
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    • pp.1329-1336
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    • 2000
  • Two experiments were carried out to investigate the effect of Murrah buffalo cows pulling sledges under field conditions on milk production and milk composition. In Exp. 1, 24 buffaloes in the fourth month of lactation were used. They were allotted to four treatments according to a $2{\times}2$ factorial arrangement: work or no work, and with or without urea-molasses cake supplementation (700 g/animal/day). Feeds consisted of 20 kg fresh elephant grass (18% DM), 2 kg rice bran per day and rice straw ad lib. The animals worked in pairs three hours per day (work done: $3464{\pm}786kJ/d$) five days a week for three months. Three teams worked in the morning and the others worked in the afternoon in the same day. The following day the working times were switched. In Exp. 2, 16 lactating Murrah buffalo cows in the sixth month of lactation were allotted to two groups (work and no work). They were fed with fresh ruzi grass (Brachiaria ruziziensis) ad lib. supplemented with 2 kg rice bran and 700 g urea-molasses cake. The working regime was similar to that of the first experiment (work done: $3753{\pm}879kJ/d$) and they worked for two months. In the first experiment, there was a small but significant drop (p<0.05) in milk yield from 3.5 to 3.0 kg/day due to work, but there was no supplementation effect. The working buffaloes lost 5.2 kg whereas the non-working animals gained 9.7 kg during the three months (p<0.05). Supplementation increased live weight by 9.9 kg as compared to -5.4 kg for those not supplemented (p<0.05). Milk composition was not affected by the treatments. In the second experiment, daily milk production was similar for both treatments and approximately 3 kg. No significant differences were found in milk composition or in live weight changes for working and non-working groups, respectively. It was concluded that work may cause a reduction in milk yield and a loss of live weight on a poor rice straw diet but that an appropriate supplementation can alleviate this situation.

Chromosomal Localization of Korean Cattle (Hanwoo) BAC Clones via BAC end Sequence Analysis

  • Chae, Sung-Hwa;Kim, Jae-Woo;Choi, Jae Min;Larkin, Denis M.;Everts-van der Wind, Annelie;Park, Hong-Seog;Yeo, Jung-Sou;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.3
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    • pp.316-327
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    • 2007
  • In this study, a Korean native cattle strain (Hanwoo) evidencing high performance in terms of both meat quality and quantity was employed in the generation of 150,000 BAC clones with an average insert size of 140 kb, and corresponding to about a 6X coverage of bovine chromosomal DNA. The BAC clones were pooled in a mini-scale via three rounds of a pooling protocol, and the efficiency of this pooling protocol was evaluated by testing the accuracy of accessibility to the positive clones, via a PCR-based screening method. Two sets of primers designed from each of two known genes were tested, and each yielded 2 or 3 positive clones for each gene, thereby indicating that the BAC library pooling system was appropriate with regard to the accession of the target BAC clones. Analyses of $3.3{\times}10^6$ base pairs obtained from the 7,090 BAC end sequence (BES) showed that 34.88% of the DNA sequence harbored the repetition sequence. Analysis of the 7,090 BES to the $1^{st}$ and $2^{nd}$ generation radiation hybrid map of the cattle genome, using the COMPASS program designed for the construction of a cattle-human comparative mapping, resulted in the localization of a total of 1,374 clones proximal to 339 $1^{st}$ generation markers, and 1,721 clones proximal to 664 $2^{nd}$ generation markers. Collectively, the BAC library and pooling system of the BAC clones from the Korean cattle, coupled with the chromosome-localized BAC clones, will provide us with novel tools for the excavation of desired clones for genome mapping and sequencing, and will also furnish us with additional information regarding breed differences in cattle.