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Comparative Study on the Viability of Frozen-thawed Primordial Germ Cells using Vitrification in Chicken Breed (초자화 동결법을 이용한 닭 품종간의 원시생식세포 동결성적의 비교)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Choi, Sung Bok;Ko, Yeoung Gyu;Do, Yoon Jung;Seong, Hwan Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.207-216
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    • 2013
  • This study was conducted to establish the method for preserving PGCs that enables long-term storage in liquid nitrogen for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of freeze-thaw treatment on viability of PGCs in chickens. PGCs were collected separately from a germinal gonad of an early embryo of 5.5~6 day (stage 28) of Isa brown, Korean Oge (KO), White Leghorn and Commercial breeds. PGCs separated from a germinal gonad of an early embryo of 5.5~6 day (stage 28) are suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and glycerol). The PGCs were then purified using magnetic activated cell sorting (MACS) method. The viability of PGCs after thawing was $87.4{\pm}0.4%$ and $89.4{\pm}0.2%$ with the 10% EG treatments with no significant difference between the Isa brown and Commercial breeds. The viability of PGCs after freeze- thawing was significantly higher for Isa brown ($87.4{\pm}0.4%$) and Commercial breeds ($89.4{\pm}0.2%$) than Korean Oge (KO) ($77.6{\pm}1.1%$) and White Leghorn ($76.2{\pm}0.9%$)(p<0.05) using 10% EG cryoprotectant. This study established a method for pre- serving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at agermplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

In Vitro Fertilization of Pig Oocytes Matured In­Vitro by liquid Boar Spermatozoa (체외성숙 돼지 난포란의 액상정액을 이용한 체외수정)

  • 박창식;이영주
    • Korean Journal of Animal Reproduction
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    • v.26 no.1
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    • pp.17-23
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    • 2002
  • The present study was carried out to investigate the effects of the maturation media such as a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium on penetrability of pig oocytes by liquid boar sperm. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. When immature pig oocytes were cultured in mTCM-199, mWaymouth MB 752/1 and NCSU-23 maturation media for 44 h in 5% $CO_2$, in air at 38.5$^{\circ}C$, the germinal vesicle breakdown (CVBD) rates of the oocytes were 95.6, 94.1 and 94.9%, respectively, and the maturation rates (metaphase II) of oocytes were 92.5, 90.1 and 91.1%, respectively. No differences were observed among the maturation media. The spermrich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ for 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes, cumulus cells were removed and oocytes (30~40) coincubated far 6 h in 0.5 $m\ell$ mTCM-199 and mTBM fertilization media with 2$\times$1061$m\ell$ sperm concentration. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ mTCM-199 and NCSU-23 culture media for further culture 6 or 42 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF, and developmental ability of oocytes at 48 h after IVF were evaluated. The oocytes in combination with NCSU-23 medium for maturation and mTBM medium for IVF increased male pronuclear formation (48.0%) compared to those in combination with mTCM-199 media for maturation and IVF, and mWaymouth MB 752il medium for maturation and mTCM-199 medium far IVF. The rates of cleaved embryos (2~4 cell stage) at 48 h after IVF were 24.1% in combination with mTCM-199 media for maturation, IVF and culture, 43.6% in combination with mWaymouth MB 75211 medium fur maturation and mTCM-199 media for IVF and culture, and 71.2% in combination with NCSU-23 medium for maturation, mTBM medium for IVF and NCSU-23 medium for culture. In conclusion, we found out the oocytes matured in vitro were fertilized by liquid boar sperm stored in BTS extender at 17$^{\circ}C$ for 5 days. We recommend the simple defined NCSU-23 medium for nuclear maturation, mTBM medium and liquid boar sperm for IVF, and NCSU-23 medium for embryo culture.

Derivation of the Synthetic Unit Hydrograph Based on the Watershed Characteristics (유역특성에 의한 합성단위도의 유도에 관한 연구)

  • 서승덕
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.17 no.1
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    • pp.3642-3654
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    • 1975
  • The purpose of this thesis is to derive a unit hydrograph which may be applied to the ungaged watershed area from the relations between directly measurable unitgraph properties such as peak discharge(qp), time to peak discharge (Tp), and lag time (Lg) and watershed characteristics such as river length(L) from the given station to the upstream limits of the watershed area in km, river length from station to centroid of gravity of the watershed area in km (Lca), and main stream slope in meter per km (S). Other procedure based on routing a time-area diagram through catchment storage named Instantaneous Unit Hydrograph(IUH). Dimensionless unitgraph also analysed in brief. The basic data (1969 to 1973) used in these studies are 9 recording level gages and rating curves, 41 rain gages and pluviographs, and 40 observed unitgraphs through the 9 sub watersheds in Nak Oong River basin. The results summarized in these studies are as follows; 1. Time in hour from start of rise to peak rate (Tp) generally occured at the position of 0.3Tb (time base of hydrograph) with some indication of higher values for larger watershed. The base flow is comparelatively higher than the other small watershed area. 2. Te losses from rainfall were divided into initial loss and continuing loss. Initial loss may be defined as that portion of storm rainfall which is intercepted by vegetation, held in deppression storage or infiltrated at a high rate early in the storm and continuing loss is defined as the loss which continues at a constant rate throughout the duration of the storm after the initial loss has been satisfied. Tis continuing loss approximates the nearly constant rate of infiltration (${\Phi}$-index method). The loss rate from this analysis was estimated 50 Per cent to the rainfall excess approximately during the surface runoff occured. 3. Stream slope seems approximate, as is usual, to consider the mainstreamonly, not giving any specific consideration to tributary. It is desirable to develop a single measure of slope that is representative of the who1e stream. The mean slope of channel increment in 1 meter per 200 meters and 1 meter per 1400 meters were defined at Gazang and Jindong respectively. It is considered that the slopes are low slightly in the light of other river studies. Flood concentration rate might slightly be low in the Nak Dong river basin. 4. It found that the watershed lag (Lg, hrs) could be expressed by Lg=0.253 (L.Lca)0.4171 The product L.Lca is a measure of the size and shape of the watershed. For the logarithms, the correlation coefficient for Lg was 0.97 which defined that Lg is closely related with the watershed characteristics, L and Lca. 5. Expression for basin might be expected to take form containing theslope as {{{{ { L}_{g }=0.545 {( { L. { L}_{ca } } over { SQRT {s} } ) }^{0.346 } }}}} For the logarithms, the correlation coefficient for Lg was 0.97 which defined that Lg is closely related with the basin characteristics too. It should be needed to take care of analysis which relating to the mean slopes 6. Peak discharge per unit area of unitgraph for standard duration tr, ㎥/sec/$\textrm{km}^2$, was given by qp=10-0.52-0.0184Lg with a indication of lower values for watershed contrary to the higher lag time. For the logarithms, the correlation coefficient qp was 0.998 which defined high sign ificance. The peak discharge of the unitgraph for an area could therefore be expected to take the from Qp=qp. A(㎥/sec). 7. Using the unitgraph parameter Lg, the base length of the unitgraph, in days, was adopted as {{{{ {T}_{b } =0.73+2.073( { { L}_{g } } over {24 } )}}}} with high significant correlation coefficient, 0.92. The constant of the above equation are fixed by the procedure used to separate base flow from direct runoff. 8. The width W75 of the unitgraph at discharge equal to 75 per cent of the peak discharge, in hours and the width W50 at discharge equal to 50 Per cent of the peak discharge in hours, can be estimated from {{{{ { W}_{75 }= { 1.61} over { { q}_{b } ^{1.05 } } }}}} and {{{{ { W}_{50 }= { 2.5} over { { q}_{b } ^{1.05 } } }}}} respectively. This provides supplementary guide for sketching the unitgraph. 9. Above equations define the three factors necessary to construct the unitgraph for duration tr. For the duration tR, the lag is LgR=Lg+0.2(tR-tr) and this modified lag, LgRis used in qp and Tb It the tr happens to be equal to or close to tR, further assume qpR=qp. 10. Triangular hydrograph is a dimensionless unitgraph prepared from the 40 unitgraphs. The equation is shown as {{{{ { q}_{p } = { K.A.Q} over { { T}_{p } } }}}} or {{{{ { q}_{p } = { 0.21A.Q} over { { T}_{p } } }}}} The constant 0.21 is defined to Nak Dong River basin. 11. The base length of the time-area diagram for the IUH routing is {{{{C=0.9 {( { L. { L}_{ca } } over { SQRT { s} } ) }^{1/3 } }}}}. Correlation coefficient for C was 0.983 which defined a high significance. The base length of the T-AD was set to equal the time from the midpoint of rain fall excess to the point of contraflexure. The constant K, derived in this studies is K=8.32+0.0213 {{{{ { L} over { SQRT { s} } }}}} with correlation coefficient, 0.964. 12. In the light of the results analysed in these studies, average errors in the peak discharge of the Synthetic unitgraph, Triangular unitgraph, and IUH were estimated as 2.2, 7.7 and 6.4 per cent respectively to the peak of observed average unitgraph. Each ordinate of the Synthetic unitgraph was approached closely to the observed one.

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Development of an Unmanned Conveyor Belt Recovery Skimmer for Floating Marine Debris and High Viscosity Oil (무인 컨베이어 벨트식 부유쓰레기 및 고점도유 회수장비 개발 연구)

  • Han, Sang-goo;Lee, Won-ju;Jang, Se-hyun;Choi, Jae-hyuk
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.23 no.2
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    • pp.208-215
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    • 2017
  • When persistent oil, such as crude oil or Bunker C oil, is spilled at sea, viscosity increases through the weathering process. Equipment that can collect this oil when mixed with floating marine debris is very limited. In this study, devices that can be attached to the outside of existing oil skimmers have been applied to the inside of the main body, to develop an unmanned conveyor belt type floating marine debris and high viscosity oil recovery skimmer, which is composed of a conveyor belt, a sweeper with a forced inflow device, and a collection tank equipped with a buoyant body. The resulting skimmer was operated at a speed of 1.2 knots at a distance of 30 m in a sea area test. It was stable when moving laterally in any direction. An oil recovery performance test was conducted using a portable storage tank, and oil was recovered from a minimum of $7.8k{\ell}/h$ to a maximum of $23.3k{\ell}/h$. Moreover, recovery of $7.7k{\ell}/h$ was obtained in a wave water tank test with floating marine debris such as PET bottles and oil mixed. If the equipment developed in this study was used in the field for oil pollution accidents, it could be expected to contribute to improved response capability. We believe our equipment could be used in further studies to improvement the performance of existing portable oil skimmers.

Development of a Biofungicide Using a Mycoparasitic Fungus Simplicillium lamellicola BCP and Its Control Efficacy against Gray Mold Diseases of Tomato and Ginseng

  • Shin, Teak Soo;Yu, Nan Hee;Lee, Jaeho;Choi, Gyung Ja;Kim, Jin-Cheol;Shin, Chul Soo
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.337-344
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    • 2017
  • To develop a commercial product using the mycoparasitic fungus Simplicillium lamellicola BCP, the scale-up of conidia production from a 5-l jar to a 5,000-l pilot bioreactor, optimization of the freeze-drying of the fermentation broth, and preparation of a wettable powder-type formulation were performed. Then, its disease control efficacy was evaluated against gray mold diseases of tomato and ginseng plants in field conditions. The final conidial yields of S. lamellicola BCP were $3.3{\times}10^9conidia/ml$ for a 5-l jar, $3.5{\times}10^9conidia/ml$ for a 500-l pilot vessel, and $3.1{\times}10^9conidia/ml$ for a 5,000-l pilot bioreactor. The conidial yield in the 5,000-l pilot bioreactor was comparable to that in the 5-l jar and 500-l pilot vessel. On the other hand, the highest conidial viability of 86% was obtained by the freeze-drying method using an additive combination of lactose, trehalose, soybean meal, and glycerin. Using the freeze-dried sample, a wettable powder-type formulation (active ingredient 10%; BCP-WP10) was prepared. A conidial viability of more than 50% was maintained in BCP-WP10 until 22 weeks for storage at $40^{\circ}C$. BCP-WP10 effectively suppressed the development of gray mold disease on tomato with control efficacies of 64.7% and 82.6% at 500- and 250-fold dilutions, respectively. It also reduced the incidence of gray mold on ginseng by 65.6% and 81.3% at 500- and 250-fold dilutions, respectively. The results indicated that the new microbial fungicide BCP-WP10 can be used widely to control gray mold diseases of various crops including tomato and ginseng.

Studies on Characteristics of Sprouting and Occurrence on paddy field of Water Chestnut(Eleocharis Kuroguwai Ohwi) (올방개 괴경(塊莖)의 맹아(萌芽) 특성(特性)과 본답(本畓)에서의 발생(發生)에 관한 연구(硏究))

  • Kim, H.D.;Park, J.S.;Park, K.Y.;Choi, Y.J.;Yu, C.J.;Rho, Y.D.;Kwon, Y.W.
    • Korean Journal of Weed Science
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    • v.16 no.4
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    • pp.264-281
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    • 1996
  • As a consequence of wide use of herbicides, Eleocharis kuroguwai Ohwi became a dorminant problem weed for rice cultivation in Korea. To understand the establishment of the weed, experiments on physio-ecological characteristics were carried out sprouting and occurrence, the results could be summarized as follows: Sprouting percentage remained 68 to 73% until the time of field emergence, indicating many of the them are still dormant. The proportion of the dormant tubers were greater for the smaller than the bigger tubers. Apical dominance was apparent in sprouting, with 84% of tuber sprouted from only one of the apical buds. Tubers sprouted from 2 or 3 buds were less than 20%, and were mostly from the bigger tubers. When the shoot growth was compared, by controlling the others, ones from apical and the next 3 buds showed similar vigorous growth, but the later ones showed poorer growth. For the longevity of tubers, deep soil storage appeared to be better than storage in temperature controlled room to 2~$3^{\circ}C$. Emergence of E. kuroguwai was better in clay soil than in sand, and the possible depth for emergence in clay soil appeared to be up to 21cm, but was 15cm in sand. When tubers were exposed to salt solutions before emergence tests, E. kuroguwai appeared to be much sensitive to salts than S.planiculmis. Among the tubers formed in previous year, 12.7% remained still viable until the end of next crop season, but with relatively strong dormancy. The first emergence was about 10 days after planting at ordinary cropping seasons, and the days to the maximum shoot number stage were 60~90 from planting. The duration was extended at early transplanting, and shoot number, leaves per shoot, and tubers developed per plant were also greater at early plantings. The 6th order offshoots were developing when E. kuroguwai was planted at early season. When planted at later seasons, generation and the number of offshoots was reduced planted at early season. When planted at later seasons, generation and the number of offshoots was reduced and the number of tubers, runner and rhizome lengths was also reduced.

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Application of Adsorption Sampling and Thermal Desorption with GC/MS Analysis for the Measurement of Low-Molecular Weight PAHs in Ambient Air (환경대기 중 저분자 PAHs 측정을 위한 흡착-열탈착-GC/MS 방법의 적용)

  • Seo, Seok-Jun;Seo, Young-Kyo;Hwang, Yoon-Jung;Jung, Dong-Hee;Baek, Sung-Ok
    • Journal of Korean Society for Atmospheric Environment
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    • v.30 no.4
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    • pp.362-377
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    • 2014
  • Polycyclic aromatic hydrocarbons (PAHs) have been of particular concern since they are present both in the vapor and particulate phases in ambient air. In this study, a simple method was applied to determine the vapor phase PAHs, and the performance of the new method was evaluated with a conventional method. The simple method was based on adsorption sampling and thermal desorption with GC/MS analysis, which is generally applied to the determination of volatile organic compounds (VOCs) in the air. A combination of Carbotrap (300 mg) and Carbotrap-C (100 mg) sorbents was used as the adsorbent. Target compounds included two rings PAHs such as naphthalene, acenaphthylene, and acenaphthene. Among them, naphthalene was listed as one of the main HAPs together with a number of VOCs in petroleum refining industries in the USA. For comparison purposes, a method based on adsorption sampling and solvent extraction with GC/MS analysis was adopted, which is in principle same as the NIOSH 5515 method. The performance of the adsorption sampling and thermal desorption method was evaluated with respect to repeatabilities, detection limits, linearities, and storage stabilities for target compounds. The analytical repeatabilities of standard samples are all within 20%. Lower detection limits was estimated to be less than 0.1 ppbv. In the results from comparison studies between two methods for real air samples. Although the correlation coefficients were more than 0.9, a systematic difference between the two groups was revealed by the paired t-test (${\alpha}$=0.05). Concentrations of two-rings PAHs determined by adsorption and thermal desorption method consistently higher than those by solvent extraction method. The difference was caused by not only the poor sampling efficiencies of XAD-2 for target PAHs and but also sample losses during the solvent extraction and concentration procedure. This implies that the levels of lower molecular PAHs tend to be underestimated when determined by a conventional PAH method utilizing XAD-2 (and/or PUF) sampling and solvent extraction method. The adsorption sampling and thermal desorption with GC analysis is very simple, rapid, and reliable for lower-molecular weight PAHs. In addition, the method can be used for the measurement of VOCs in the air simultaneously. Therefore, we recommend that the determination of naphthalene, the most volatile PAH, will be better when it is measured by a VOC method instead of a conventional PAH method from a viewpoint of accuracy.

RHEOLOGICAL CHARACTERIZATION OF COMPOSITES USING A VERTICAL OSCILLATION RHEOMETER (수직 진동형 Rheometer를 이용한 복합레진의 유변학적 성질의 측정)

  • Lee, In-Bog;Cho, Byung-Hoon;Son, Ho-Hyun;Lee, Sang-Tag;Um, Chung-Moon
    • Restorative Dentistry and Endodontics
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    • v.29 no.6
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    • pp.489-497
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    • 2004
  • Objective: The purpose of this study was to investigate the viscoelastic properties related to handling characteristics of composite resins, Methods: A custom designed vertical oscillation rheometer (VOR) was used for rheological measurements of composites. The VOR consists of three parts: (1) a measuring unit, (2) a deformation induction unit and (3) a force detecting unit, Two medium viscous composites, Z100 and Z250 and two packable composites, P60 and SureFil were tested. The viscoelastic material function, including complex modulus $E^{*}$ and phase angle ${\delta}$, were measured. A dynamic oscillatory test was used to evaluate the storage modulus (E'), loss modulus (E") and loss tangent ($tan{\delta}$) of the composites as a function of frequency ($\omega$) from 0.1 to 20 Hz at $23^{\circ}C$. Results: The E' and E" increased with increasing frequency and showed differences in magnitude between brands. The $E^{*}s$ of composites at ${\omega}{\;}={\;}2{\;}Hz$, normalized to that of Z100, were 2.16 (Z250), 4,80 (P60) and 25.21 (SureFil). The magnitudes and patterns of the change of $tan{\delta}$ of composites with increasing frequency were significantly different between brands. The relationships between the complex modulus $E^{*}$, the phase angle ${\delta}$ and the frequency \omega were represented by frequency domain phasor form, $E^{*}{\;}(\omega){\;}={\;}E^{*}e^{i{\delta}}{\;}={\;}E^{*}{\angle}{\delta}$. Conclusions: The viscoelasticity of composites that influences handling characteristics is significant different between brands, The VOR is a relatively simple device for dynamic, mechanical analysis of high viscous dental composites. The locus of frequency domain phasor plots in a complex plane is a valuable method of representing the viscoelastic properties of composites.

Development of the Automatic Feeder for Growing-finishing Pigs (육성비육돈용 자동급이기 사료공급장치 개발에 관한 연구)

  • Yoo, Y.H.;Song, J.I.;Choi, H.C.;Kim, J.H.;Park, K.H.;Kang, H.S.;Chang, D.I.
    • Journal of Animal Environmental Science
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    • v.15 no.3
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    • pp.241-250
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    • 2009
  • This study was conducted to develop an prototype automatic feeder (AF) for growing-finishing pigs. The main components of AF were a feed storage hopper, a feeding motor, a feed agitator, a control box and a programmable IC, which were controlled by a personal computer. The powder type feed transfer rate of AF was average $9.83{\pm}0.4\;g\;s^{-1}$. In feeding test, growing pigs (Landrace) of about 43 kg live weight were used in the study, and was fed over a 6 weeks in pens with solid concrete floors. For feeding trials with AF, the operation time of the feeding motor was set to 2, 3, 4, 5, and 6 seconds per feeding. Pigs frequently used AF from 05:00 to 11:00 and from 11:00 to 17:00 without relationship to the operation time of the feeding motor. The AF operation time of the feeding motor to minimize feed loss was between 2 and 4 seconds. Pigs fed with AF had same or slightly higher average daily gam (0.8~0.9 kg) than that with a commercial feeder, and average daily feed intake (2.76~2.93 kg) and feed conversion ratio (3.10~3.66) of pigs fed with AF were same or lower than those with the commercial feeder except the operation time of the feeding motor set to 6 seconds. As a result, AF would help to use and improve the productivity of growing-finishing pigs.

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Field Survey on Liquid Manure Utilization in the Agricultural Farms (경종농가에서의 액비이용 실태조사)

  • Choi D. Y.;Kwag J. H.;Park C. H.;Jeong K. H.;Jeon B. S.;Choi H. C.;Kang H. S.;Yang C. B.;Choi H. L.
    • Journal of Animal Environmental Science
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    • v.10 no.3
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    • pp.155-162
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    • 2004
  • The livestock liquid manure is one of important source for production of friendly environmental crops and have been used widespreadly in recent years. This survey is to eventually investigate the actual conditions of liquid manure utilization for cultivation of crops in the agricultural farm, based on the survey for 61 selected farms in 8 provinces(except Jeju province) included 22 counties in Korea. The results obtained in this survey were summarized as follow; $72.1\%$ of liquid manure storage tank(44) was located in the farmland and $27.9\%$(17) was in the fm. Most of liquid manure tank volume and material were 200 M/T($67.2\%$) and Polyethylene Double Frame panel($44.3\%$). The pro-portions of liquid manure application land were $45.9\%$ for rice paddy, $36.1\%$ for dry field, $16.4\%$ for orchard and $1.6\%$ for other, respectively. The controversial points of liquid manure utilization were malodor($60.7\%$), equipment possession($22.9\%$), no problem($13.1\%$) and farmland possession ($2.3\%$), respectively.

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