• 환경위생공학
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• 제15권4호
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• pp.105-113
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• 2000

We have isolated 65 strains(2.0%) of Y. enterocolitica among 3,219 water samples from 380 spring water sites in Seoul from 1994 to 1999. The biochemical characteristics of isolated strains revealed that TSI was A/A, urea, M.R.($37^{\circ}C$), nitrate, motility($37^{\circ}C$), sorbitol, maltose, manitol, arabinose, mannose, trehalose, xylose were positive(100%) and H$_2$S, arginine, lysine, oxidase, citrate, V.P.($37^{\circ}C$), DNase, motility($37^{\circ}C$), dulcitol, adonitol, lactose and raffinose were negative(100%). In in vitro virulence test, positive rate of AAG and CRMOX were 9.2% and 4.6%, respectively. However in the virulence gene detectable gene detectable test by multiplex-PCR using ail, yst, virF genes, 65 strains were all negative, meaning that Y.enterocolitica strains from domestic spring water were not detected for the virulence. Otherwise, mutiplex-PCR which using ail, yst and subgenus-specific primer pair was the best for identifying the virulence of Y. enterocolitica.

• 대한치과보철학회지
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• 제49권2호
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• pp.138-144
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• 2011

연구 목적: 본 연구는 최근 개발된 알칼리 이온수 e-WASH의 의치에 부착된 미생물 의치세정 효과에 대해 위상차현미경을 이용한 미생물 관찰법을 통하여 알아보고자 하였다. 연구 재료 및 방법: 실험군 41개, 대조군 26개의 의치를 무작위로 선정하여 실험군에서는 알칼리 이온수e-WASH로, 대조군에서는 수돗물에 5분간 침지 전, 후 의치 내면의 치면세균막을 채취하였다. 위상차현미경으로 구강미생물의 형태적, 종류별 양과 운동성에 대하여 세정 전, 후별 구강미생물의 상태를 관찰하였다. 결과: 1. 알칼리 이온수 e-WASH로 의치를 세정한 실험군에서는 세정 전보다 세정 후 구균의 양, 간균의 양, 간균의 운동성, 실사균의 양, 실사균의 운동성, 콤마/나선균의 양, 콤마/나선균의 운동성 등 모든 조사항목에서 감소하였다 (P<.05). 한편 대조군에서는 구균의 양은 감소하였으나 (P<.05), 다른 조사항목에서는 모두 세정 전, 후 간에 차이가 없었다. 2. 알칼리 이온수e-WASH로 의치를 세정한 실험군에서는 구균의 양, 간균의 양, 간균의 운동성, 실사균의 양, 실사균의 운동성, 콤마/나선균의 양, 콤마/나선균의 운동성 등 모든 조사항목에서 수돗물로 의치를 세정한 대조군보다 각 구강미생물의 양과 운동성이 적게 나타났다 (P<.05). 결론: 위의 결과를 통해 알칼리 이온수 e-WASH가 의치표면의 각종 미생물의 양과 운동성을 효과적으로 감소시키므로 의치세정제로 추천된다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권12호
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• pp.1716-1721
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• 2006

The present study was conducted to observe the effect of osmolality of glycerolated TEST-yolk glycerol extenders on post-thawing sperm kinematics of ram spermatozoa of the native Malpura breed maintained in a semi-arid tropical environment. Good quality semen obtained from adult rams was pooled, split and diluted to 1,000 million spermatozoa per ml in complete TEST-yolk-glycerol extenders of 900, 1,200, 1,500 and 1,800 mOsm/kg osmolality. Diluted semen samples were loaded in 0.25 ml straws and cooled down to $-125^{\circ}C$ freezing temperature at the rate of $-25^{\circ}C$ per minute under controlled conditions before plunging into liquid nitrogen for storage. The thawing of straws was performed at $50^{\circ}C$ in a water bath for 10 seconds and sperm kinematics of the frozen-thawed spermatozoa were assessed by a computer-assisted sperm analysis technique. Osmolality of diluent had no significant effect on post-thawing % motility, % rapid, % medium and % slow moving frozen-thawed spermatozoa but significantly (p< 0.05) affected the % linearity and % straightness. The post-thawing % motility and % rapid motile spermatozoa were highest in samples extended in diluent of 1,500 mOsm/kg osmolality and lowest in 900 mOsm/kg. The curvilinear velocity of spermatozoa was significantly (p<0.05) higher for samples extended in 1,800 mOsm/kg, compared to those in 900 and 1,200 mOsm/kg, but the effect was not significantly different to those extended in diluent of 1,500 mOsm/kg osmolality. The study indicated that ram spermatozoa could tolerate a wide osmolality range for dilution in the complete TEST-yolk-glycerol extender for their cryosurvival. The highest recovery of motile spermatozoa following thawing was achieved in samples extended in the TEST-yolk-glycerol diluent of 1,500 mOsm/kg osmolality.

• 한국가축번식학회지
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• 제25권4호
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• pp.409-419
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• 2001

본 실험은 PF과 FPP가 소와 사람 동결 정자에 첨가되었을 때 응해 후 정자의 체외 생존성, 운동성 그리고 intact acrosome 향상에 기여할 수 있는지의 여부를 조사하고자 실시하였다. 사람의 정액은 TYB 동결 배양액을 사용하여 초 급속 동결하였다 PF과 FPP 첨가 효과는 각각의 시약이 동결-응해된 소나 사람 정자의 현미경적 조사에 의한 운동성에 미치는 영향과 Coomassie brilliant blue 염색방법에 의한 intact acrosome의 비율에 미치는 영향으로 조사하였다. Bovine 동결 응해 정자에 PF을 첨가하여 운동성을 조사하였던 바, 5 mM 처리군 (50.0%)이 대조군 (34.0%) 보다 유의하게 높은 운동성을 보여주었다 (F＜0.05). 동결 응해된 소 정자에 FPP를 농도에 따라 처리하여 intact acrosome을 조사하였던 결과, 50 nM 치리춘 (49%)이 대조군과 25 nH 처리군 (30.0, 38.0%) 보다 유의하게 많은 intact한 acrosome을 유지하였다 (P＜0.01). 사람 정자에서 동결에 앞서 PF을 농도에 따라 첨가하여 동결 응해 후 운동성을 조사한 결과, 5.0 mM 처리군 (51.0%)이 대조군과 2.5 mM (39.0, 40.0%) 처리군의 운동성보다 높았다 (P＜0.01). 사람 정액의 모든 동결 처리과정 (동결전, 동결, 응해후)에서 50 nM (75.5%) FPP 첨가가 intact acrosome percentage 유지하는데 유의한 효과가 있었다 (대조군: 45.0; 25 nM: 53.0; 100 nM: 68.0%) (P＜0.01). PF와 FPP 첨가하여 사람 정자의 동결융해 후 운동성과 intact acrosome에 미치는 영향을 동시에 비교해본 결과, 운동성에서는 PF 처리군이 약간 높지만 intact acrosome rate는 FPP 처리군의 결과 (65.0%)가 PF 처리군 (43.0%)보다 유의하게 높았다 (P＜0.05). 따라서 본 실험은 동결-융해된 소 정자에 PF이나 FPP 첨가는 정자의 운동성이나 intact acrosome 비율을 좀더 개선시킬 수 있고, 특히 사람 정자는 동결 전 과정에 FPP를 첨가하는 것이 정자의 체외 생존성, 운동성 그리고 intact acrosome을 유의하게 향상시킬 수 있다는 것을 시사한다.

• Toxicological Research
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• 제25권4호
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• pp.203-207
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• 2009

Present study was conducted to investigate potential effects of epichlorohydrin on testicular and epididymal function in male rats. The test chemical was administered to adult male rats by gavage at dose levels of 0, 3.125, 12.5, and 50 mg/kg/day for 7 days. Testicular and epididymal function were assessed by measurement of reproductive organ weight, testicular spermatid count, epididymal sperm count, motility and morphology, and histopathology in rats. At 50 mg/kg, a decrease in the sperm motility and an increase in the incidence of sperm abnormalities were observed. Histopathological examinations revealed an increase in the incidence of histopathological changes including cell debris in the ducts, vacuolization of the epithelial cells, oligospermia, and epithelial disruption in the proximal caput epididymidis. At 12.5 mg/kg, an increase in the incidence of histopathological changes of the epididymidis was found. There were no treatment-related effects at 3.125 mg/kg. These results show that 7-day repeated oral administration of epichlorohydrin to male rats results in adverse effects on sperm motility, sperm morphology, and epididymal histology at $\geq$ 12.5 mg/kg/day.

• Clinical and Experimental Reproductive Medicine
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• 제29권2호
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• pp.149-157
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• 2002

Objectives: To assess the scavenging effect of carnitine derivatives on oxidative damage to sperm during sperm processing, cryopreservation and thawing. Materials and Methods: Fresh semen samples from 20 normal healthy volunteers were collected by masturbation after at least 48 hours abstinence. After liquefaction of semen samples at room temperature, the specimens were diluted with sperm wash media (Ham's F-10, Life technologics) to a uniform density of $20{\times}10^6/ml$. L-carnitine or acetylcarnitine were added with various concentration of $0{\mu}M$, $10{\mu}M$, $30{\mu}M$ in semen sample or cryoprotectant. All specimens were cryopreservated at $-196^{circ}C$ $LN_2$ for 3 days. Sperm motility, vitality, fertilizing capacity, reactive oxygen species formation and the level of lipid peroxidation were analyzed by computer assisted semen analyzer, eosin-nigrosin stain, hypoosmotic swelling test, chemiluminescence and thiobarbituric acid method, respectively, during sperm processing, cryopreservation and thawing. Results: The sperm motility was only increased in proportion to the concentration of acetylcarnitine with no statistical significance (p>0.05). The sperm vitality was also significantly improved in proportion to the concentration of acetylcarnitine with statistical significance (p<0.05). The sperm fertilizing capacity was significantly increased in proportion to the concentration of L-carnitine and acetylcarnitine and reactive oxygen species generation and lipid peroxidation were significantly decreased with same fashion (p<0.05). On comparison of effects between L-carnitine and acetylcarnitine, acetylcarnitine was superior to L-carnitine on the improvement of sperm motility and vitality as well as the suppression of reactive oxygen species generation and lipid peroxidation. Conclusions: These results suggest that carnitine derivatives have a scavenging effect against oxidative damages during sperm processing, cryopreservation and thawing. Therefore, carnitine derivatives may be useful as an oral antioxidant in patients with male infertility due to increased ROS generation.

• Asian-Australasian Journal of Animal Sciences
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• 제17권11호
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• pp.1501-1508
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• 2004

In this study the effect of extenders and temperatures on sperm viability and fertilizing capacity of boar sperm during long-term storage was investigated. Acrosomal integrity, membrane integrity, motility and hypo-osmotic resistance were evaluated by fluorescence and light microscopy. An in vitro fertilization test was performed to assess the fertilizing capacity of stored spermatozoa. The five diluents tested were ranked according to their ability to maintain sperm functional parameters and Zorlesco (ZO) extender with BSA or with PVA instead of BSA produced the best results. Zorlesco extender substituted with PVA (ZO+PVA) was found to maintain motility both at 15 and 20$^{\circ}C$. within 5 days of storage, but the quality of semen stored at 15$^{\circ}C$ decreased thereafter as compared to semen stored at 20$^{\circ}C$ Semen stored at 5$^{\circ}C$ demonstrated rapid loss of motility already within 24 h. Both fertilization and cleavage of semen stored at 20$^{\circ}C$ in ZO substituted with PVA instead of BSA did not change significantly until day 8 of storage. It is therefore concluded that PVA can be used to substitute for BSA and 20$^{\circ}C$ was more suitable than 15$^{\circ}C$ for boar semen storage, and in vitro fertilizing capacity of spermatozoa was maintained for at least 8 days in ZO+PVA at 20$^{\circ}C$.

• 한국가축번식학회지
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• 제10권1호
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• pp.91-99
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• 1986

This experiment was undertaken to examine the effects of HIS treatment on the motility and acrosome reaction of frozen bovine spermatozoa and to test their abilities to interact with zona-free hamster eggs in vitro. Also, in vitro results were compared with those of bull's fertility in AI. The frozen semen from four Holstein bulls were exposed to HIS-DM for 5 minutes after thawing and then preincubated for 60 minutes in DM prior to insemination. The hamster eggs were mounted, fixed and stained 6 hours after exposure to boving spermatozoa and examined under a phase-contrast microscope. 1. The sperm motility expressed as a mobility index dro, pp.d significantly from 60-75 to 12-24 after exposure to HIS-DM, but increased in 32 to 41 at insemination. Bull C showed a low motility index than those of the orher bulls. The percentage of acrosome reaction by staining procedure were increased by HIS-DM treatment but did not change during 7 hours incubation period in DM. 2. The overall percentage of hamster eggs interacting with bull spermatozoa was 56.3%, 58.3%, 66.6% and 70.0%, respectively. Although there was no significant difference among bulls in the penetration rate of spermatozoa into hamster eggs, high proportions of eggs interacted with spermatozoa from Bull C and D than those from Bull A and B. 3. The conception rates (60-90 day RP) resulting from AI were 62.5%, 67.5% and 70.9% for Bull A, B and C, respectively. These results were in good agreement with the invitro results that the proportions of bull sperm-egg interction were greater for Bull C than for Bull A and B.

• 한국수정란이식학회지
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• 제24권4호
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• pp.275-279
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• 2009

The beneficial effect of glycerol as a cryoprotectant, especially for sperm cryopreservation, has been shown in many studies. However, glycerol is toxic to living cells, and boar sperm in particular show greater sensitivity to glycerol than sperm from other domestic animals. Amides have been studied as alternative cryoprotectants for freezing stallion sperm. Sperm frozen in methylformamide or dimethylformamide as cryoprotectants show similar motility when thawed compared with sperm frozen in glycerol. We evaluated the cryoprotective effects of dimethylformamide on boar sperm freezing. To test the effect of amides, the concentration of boar semen was adjusted to $10^9sperm/mL$, and seminal plasma was removed using Hulsen solution. After centrifugation, the pellet was diluted in modified-Modena B extender. Lactose-egg yolk (LEY) extender was used as the cooling extender. The freezing extender was madeed aaddition of the optimal amount of glycerol and amides to LEY-Glycerol-Orvus ES Paste extender, and this extender was used for the second dilution. Diluted sperm were frozen in liquid nitrogen using the 0.5 mL straw method. Sperm frozen in extender with glycerol as a cderol were compared with those frozen in extender including the different amides. Sperm were tested for motility, viability, the sperm chromatin structure assay, and normal apical ridge after thawing. The percent of motile sperm diluted in glycerol was as high as that in the stallion study (61%). Dimethylformamide showed positive effects on sperm quality and was better than glycerol. Methylformamide provided similar sperm quality as glycerol. Therefore, dimethylformamide is useful for reducing cryoinjury in boar sperm and is expected to be useful as an alternative cryoprotectant.

• Animal Bioscience
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• 제36권10호
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• pp.1530-1535
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• 2023

Objective: Semen cryopreservation result in decreased sperm parameters and fertilization ability. Sericin exhibits antioxidant activity by reducing lipid peroxidation resulting from free radicals, which can potentially improve cryopreservation outcomes. The present study aimed to examine the efficacy of various sericin concentrations supplemented with a rooster semen-freezing extender on post-thaw semen quality and fertilizing ability of sperm after cryopreservation. Methods: Semen samples were collected from 40 roosters (5 reps), then were pooled, and divided into four groups by the levels of sericin supplementation (0%, 0.25%, 0.50%, and 0.75%) in a freezing extender. Semen suspensions were loaded in medium straw (0.5 mL) and cryopreserved with the traditional liquid nitrogen vapor method. Post-thawed semen was evaluated for sperm motility, sperm viability, and lipid peroxidation. Also, the fertility test was determined. Results: The results showed that supplementation of the freezing extender with 0.50% to 0.75% sericin resulted in greater total motility and progressive motility and lower malondialdehyde levels than the other groups after cryopreservation (p<0.05). However, the viability of 0.75% decreased compared with the value of 0.50% sericin supplementation (p<0.05). Moreover, the fertility and hatchability of total eggs were significantly higher in the 0.50% sericin group than in the other groups (p<0.05). Conclusion: In conclusion, 0.50% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved rooster semen.