• Journal of Animal Science and Technology
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• v.44 no.4
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• pp.483-490
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• 2002

The aim of this study was to determine if a low-molecular weight casein hydrolysate has an anti- hypertensive effect in spontaneously hypertensive rats (SHR). Prior to the in vivo experiment, the casein hydrolysate was confirmed to be resistant to gastrointestinal digestion by confirming the retention of its potency as an inhibitor of angiotensin I-concerting enzyme after incubation with pepsin, trypsin, or chymotrypsin. The in vivo anti-hypertensive effect of the hydrolysate was determined by the tail cuff method. Following an oral administration of the hydrolysate solution, the systolic blood pressure (SBP) decreased by 12.9% (－28.9mmHg; P<0.05) at 3 h after the administration at a dose of 500mg/kg body weight. When the hydrolysate was administered as an emulsion with 30% egg yolk, its anti-hypertensive effect was even more greater at the same dose(－30.8mmHg or －15.9%; P<0.01). In a 50-day long-term trial where the casein hydrolysate was administered once a day, the SBP-lowering effect of the hydrolysate was apparent (P<0.05) from day 35 through the end. Moreover, organ weights and plasma glutamate oxaloacetate transaminase and glutamate pyruvate transaminase activities of the administered SHR were not significantly different from those of controls at the end of the long-term trial.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.16 no.4
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• pp.389-394
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• 1983

The right eye flounders, Limanda yokohamae Gunther widely distributed in the coastal waters of Korea and Japan. On Feburuary 3, 1983, the authors obtained a number of artificial fertilized eggs from the adult fishes(male: 285mm in total length; female: 297mm) caught by a trawl. The eggs of this species is demersal and adhesive, and the diameter of these eggs was varied in $0.71{\sim}0.80mm$. The egg capsule is colorless and transparent, and the eggs do not contain any oil globules, The hatching took place in 120 hour after fertilization at the water temperature $5.5{\sim}17.0^{\circ}C$. The newly hatched larvae were $2.64{\sim}2.72mm$ in total length with long trunk. Myotome number was $9{\sim}10+30{\sim}32=39{sim}42$ and yellowish brown melanophores were appeared. In 3 days after hatching out. The larvae attained 3.3mm in total length, and the mouth began to move. Xanthophore appeared also on the opereulum at this time. After 7 days the larvae attained 3.70mm in total length, and became the postlarvae absorbing the yolk completely.

• Applied Biological Chemistry
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• v.38 no.4
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• pp.320-324
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• 1995

Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.

• Journal of Veterinary Clinics
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• v.16 no.2
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• pp.375-380
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• 1999

This study was conducted to develop an intrauterine inseminator (IUI) to deposit of frozen semen into uterus and to evaluate the results obtained after artificial insemination by IUI. Two Japanese spitzs (2 to 4 years of age) were used as semen donors. Semen was collected by manual masturbation into sterile glass collection tubes and separated into 3 fractions with only the sperm-rich fractions retained for further examination. Sperm motility >70%, sperm concentration of 200 to $400{\times}10^6 cells/ml$$\times$g for 5 min and poured out the suspended solution, and then diluted with 2 ml Tris-buffer which was consisted of 2.4 g Tris, 1.4 g citric acid, 0.8 g glucose, 0.1 $\mu\textrm{g}$/ml streptomycin, 100 IU/ml penicillin, 20 ml egg yolk to 100 ml mili-Q water (Ext I) or supplemented with 8 ml glycerol and 1 ml Equex STM paste to 100 rnl (Ext II). The diluted semen was cooled to 5$^{\circ}C$ in cold room, where the temperature in the sample reached 5$^{\circ}C$. Two h after beginning the cooling procedure, 2 ml of Ext II, also at 5$^{\circ}C$, was added and mixed by gently reversing the tubes several times during 1 h. The final sperm concentration for freezing was approximately $50{\times}10^6 cells/ml$. After equilibration, the semen was loaded into 0.5 ml straw and frozen on the liquid nitrogen vapour in styrofoam box. The straws were thawed at 7$0^{\circ}C$ for precisely 6 sec. After thawing of each straw, the frozen semen can survived over 50% motility. All the females were inseminated twice with 1 ml of $25{\times}10^6 cells/ml$

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.2
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• pp.164-167
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• 2004

This study was carried out to investigate in vitro fertilization and development of in vitro matured pig oocytes inseminated with the Duroc boar sperm by different sperm washing media after thawing of the 5 ml frozen straws. Immature follicular oocytes (30-40) were transferred into each well of a Nunc 4-well multidish containing $500{\mu}l$ mTCM199 maturation medium. The sperm rich portion of ejaculates was collected into a 250 ml insulated vacuum bottle and gradually cooled 22 to $24^{\circ}C$ over a 2 h period. Semen was centrifuged at 800 g for 10 min and the seminal plasma discarded. Sperm were esuspended in a lactose-egg yolk and N-acetyl-Dglucosamine (LEN) diluent to contain $1{\times}10^{9}$ sperm/ml and cooled to $5^{\circ}C$ over a 2 h period. Immediately before freezing, semen was rediluted with an equal volume of LEN+4% glycerol and packed into 5 ml straws. After thawing of the 5 ml straw, the 5 ml semen was diluted with 20 ml Beltsville thawing solution (BTS) at room temperature. Oocytes were inseminated with untreated (unwashed and nonpreincubated) or treated sperm (washed two times in BTS, mTLP-PVA and mTBM media, respectively and nonpreincubated) with $2{\times}10^{7}$ sperm concentration. Oocytes were coincubated for 6 h in $500{\mu}l$ mTBM fertilization. At 6 h after IVF, oocytes were transferred into $500{\mu}l$ NCSU-23 culture medium for further culture of 6 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. Sperm penetration rate, male pronuclear formation and rate of cleaved embryos were higher in the BTS, mTLP-PVA and mTBM treatments than the unwashed treatment (p<0.05). The rate of blastocysts from the cleaved oocytes (2-4 cell stage) were higher in the mTLP-PVA treatment than in the unwashed, BTS and mTBM treatments. In conclusion, we recommend the washing of frozen-thawed sperm with mTLP-PVA medium before in vitro fertilization of oocytes in mTBM medium.

• Korean Journal of Food Science and Technology
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• v.45 no.6
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• pp.791-795
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• 2013

The aim of this study was to compare the performance of a standard culture method and polymerase chain reaction (PCR) for the detection of Staphylococcus aureus (S. aureus) in milk and meat products. Milk, dried infant formula, sausage and ground beef that had been artificially inoculated with S. aureus were enriched in tryptic soy broth. After the enrichment, a loopful was inoculated onto Baird-Parker agar with egg-yolk-tellurite. In parallel, 23S rRNA was amplified by PCR from samples of the enriched broth. Suspected S. aureus colonies grown on selective agars were finally confirmed by a coagulase test and colony PCR. No significant statistical differences were observed between the incidence of S. aureus detected by the culture method and the incidence detected by PCR, in milk or dried infant formula. However, in sausage and ground beef, the number of positives detected by PCR was significantly higher than by the culture method (p<0.05). Our findings suggest that PCR could be an effective screening tool for the detection of S. aureus compared to the standard culture method.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.90-90
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• 2002

The purpose of this study was to investigate the effect of kind and combination of sugars on the viability and acrosome damage of post-thaw spermatozoa in canine. The extender used was Tris-citric acid extender (Tris-buffer) supplemented with 20% Egg-yolk, 8% glycerol, 1% Equex STM paste, and 70 mM sugars such as monosaccharide (fructose and xylose) and disaccharide(trehalose). To evaluate of sugar combination, the sugars supplemented in Tris-buffer were combined such as single (fructose, xylose, trehalose), two combinations (Fruc+Tre, Fruc+xyl, Tre+xyl) and three combinations (Fruc+Tre+Xyl). The concentration of sperm collected were adjusted of 50${\times}$10$\^$6/ per straw for freezing. The frozen spermatozoa were thawed at 37$^{\circ}C$ for 1 min and then analysis for CASA program in Livestock Improvement Main Center, NACF. The motility of post-thaw spermatozoa in Fruc+Tre was higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (79％ vs. 63, 66, 70, 71, 74 and 75%). The progressive motility after CASA analysis in Fuc+Tre group was also higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (67% vs. 53, 57, 60, 61, 62 and 64%). The acrosome damage of post-thaw spermatozoa stained was not significantly different among treatment groups such as fructose, trehalose, xylose, Fru+Tre, Fru+xyl, Tre+xyl and Freu+tre+xyl (17.7, 18.3, 28.0, 17.0, 19.7, 20.0 and 19.0%). The results indicated that the motility and progressive motility of post-thaw spermatozoa in Fru+Tre group was better, and acrosome normality was not different among all groups. The use of Tris-buffer supplemented with Fru+Tre as sugar for frezing of canine spermatosoa could be better and apply to semen banking and artificial insemination.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.193-198
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• 2013

This study was designed to evaluate the effect of L-cysteine on sperm characteristics and oocyte cleavage in vitro in Korean native cattle. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 5, 10 and 20 mM L-cysteine before cryopreservation. The viability in frozen-thawed sperm were estimated by SYBR14/PI double stain, acrosome damage with FITC-PNA, mitochondria intact with Rhodamin123 and hydrogen peroxide($H_2O_2$) level with carboxy-DCFDA by flow-cytometry. The developmental capacity was also assessed with cleavage rates in oocytes fertilized in vitro by frozen-thawed sperm. In results, the sperm viability was significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). In addition, acrosome damage was significantly decreased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). The mitochondria intact was also significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). On the other hand, the cleavage rates were significantly increased in 0 mM, 5 mM and 10 mM groups than 20 mM concentration of L-cysteine (p<0.05). The oocyte degeneration of oocytes were significantly decreased in 0 mM, 5 mM and 10 mM groups than in 20 mM L-cysteine group (P<0.05). However, there are no significantly differences among the L-cysteine treatment groups. We suggest that concentration of 10 mM L-cysteine have beneficial impact for sperm cryopreserved in Korean native cattle. This result also could be recommended for artificial insemination program if supported by an improvement in the fertility results and required further study.

• Korean Journal of Ichthyology
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• v.13 no.1
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• pp.74-84
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• 2001

The early developmental stages, growth and morphological changes of the Korean bullhead, Pseudobagrus fulvidraco, were studied from a series of reared specimens. Details of the early developmental stages are illustrated with special reference to morphological transformations. Egg and sperm of Korean bullhead were obtained from mature adults under hormonal treatment, fertilized artificially, and incubated in the aquarium. The incubation period of fertilized eggs was 55 to 66 hours at a temperature of 24.9${\pm}$0.34$^{\circ}$. Larvae were fed successively with Artemia salina and Daphnia magna for 2 to 15 days and artificial food after 20 days. Fertilized eggs were adhesive and spherical with a diameter of 2.04mm(n = 100). The mean total length of newly hatched larvae was about 4.92${\pm}$0.33 mm. Mouth opening occurred on one-day-old yolk-sac larvae, and initial feeding was observed on the third day after hatching. The morphological transitions from larvae to juvenile and juvenile to young stages occurred when the fish reached about 17 mm in total length (about 13days after hatching) and about 32 mm in total length (about 30 days after hatching), respectively. Many changes in proportion of body parts to total length were observed at about 7~8 mm and 30~32 mm, corresponding to the transformations from larvae to juvenile and from juvenile to young, respectively. In comparing relative growth of each body part against total length, those characteristics related to head parts showed positive growth in the pre-larval stages, while those concerning mobile abilities showed positive growth in the post-larval stage.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.9
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• pp.1247-1255
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• 2016

Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.