• 한국수정란이식학회지
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• 제28권2호
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• pp.113-119
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• 2013

The study focuses on the quality assessment of Black Bengal buck semen preserved at chilled condition. In this in vitro trial, collected semen from Black Bengal bucks was preserved at chilling temperature ($4{\sim}5^{\circ}C$) in tris-glucosecitrate yolk medium of 1:5 ratios for four days. Artificial Vagina (AV) method was utilized to collect semen from buck. General evaluation of semen includes the color, mass activity and density were measured by direct visual examination. However, computer-assisted sperm analysis (CASA) and phase contrast microscopy were used to figure out the motility (%), hyper-activated (HYP) motility (%) and number of abnormal spermatozoa (%) initially, and at every 24 h intervals. The result revealed that spermatozoa preserved at chilling temperature showed significantly (P<0.05) lower motility and HYP motility with the progression of preservation. The number of phenotypically abnormal spermatozoa significantly (P<0.05) increased following preservation. Although significant positive correlation (r=0.945; P<0.05) was existed between % motile and % HYP motile spermatozoa however, the % of morphologically abnormal spermatozoa was negatively correlated with % motile (r=-0.997; P<0.05) and % HYP motile spermatozoa (r=-0.946; P<0.01). Therefore, we concluded that the quality of chilled semen progressively losses its viability and doesn't remain useable after certain period of preservation with respect to its motility and morphology.

• 동의생리병리학회지
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• 제17권1호
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• pp.113-117
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• 2003

This study was performed to investigate the anti-depression effect of herbal extracts (Rehmanniae Radix Preparat, Corni Fructus, Lycii Fructus, Pinelliae Rhizoma, Nelumbinis Semen) in the chronic mild stressed rat. After adaptation period for 2 weeks, each herbal extract was administered to the rats divided 5 groups. For comparison, St. John's wart was administered to another group in the same environment. The chronic mild stress was induced to rats by overnight illumination for 2 days before the forced swimming test. Struggling time, first latency and rest duration were calculated during the forced swimming test to evaluate the anti-depressant effect. The results were as follows: 1. In the result of evaluation of the struggling time, there was no significance in Pinelliae Rhizoma (-1.1%), Corni Fructus (-3.9%), Rehmanniae Radix Preparat (15.2%), Lycii Fructus (4.9%), St. John's wart (25.2%). There was a significant increase only in Nelumbinis Semen (43.9%) (P=0.0037). 2. In the result of evaluation of the first latency time, there was no significance in Lycii Fructus (-21.37%), Rehmanniae Radix Preparat (38.4%), Pinelliae Rhizoma (65.5%), Corni Fructus (29.2%), St. John's wart (75.8%). There was a significant increase only in Nelumbinis Semen (90.2%) (P=0.0116). 3. In the result of evaluation of the first rest duration, there was no change in St. John's wart, and Decrease was shown in Pinelliae Rhizoma (62.4%), Corni Fructus (31.6%) Rehmanniae Radix Preparat (63.1 %), Lycii Fructus (12.4%), Nelumbinis Semen (59.0%), but there was no significance in all. 4. Nelumbinis Semen is considered to have a superior anti-depression effect than the other herbal extracts, and to have a superior and-depression effect than St. John's wart being used commonly.

• Toxicological Research
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• 제25권3호
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• pp.125-131
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• 2009

Korea is representative of a country that consumes herbal medicines; most of the herbal medicines circulating in South Korea have been imported from developing countries in Southeast Asia, such as China and Indonesia. Recently, domestic hygiene and safety are issues that have come to the forefront, because herbal medicines currently in circulation could possibly contain contaminants or residues. Furthermore, the appearance or discovery of harmful new species due to environmental and industrial developments is becoming a social problem. Therefore, it may be necessary to consider and investigate these matters on a continual basis. Recently, mycotoxin contaminations in such foods as cereals, nuts, and powdered red pepper have been reported. They have become a problematic issue; the possibility of contamination in herbal medicines has also been considered. Nevertheless, recognition of and research into mycotoxin contamination in herbal medicines has been scarce because herbal medicine is used in only a few nations. In this research, we identified contamination by aflatoxin which is known to be the most potent mutagenic, carcinogenic, and teratogenic mycotoxin in Thujae Semen, a herbal medicine. We also found co-contaminations involving other mycotoxins, including ochratoxin A and zeraleanone.

• Asian-Australasian Journal of Animal Sciences
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• 제21권4호
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• pp.494-498
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• 2008

MTT (3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) reduction assay is a method that validates the viability of an active cell. Dehydrogenase in mitochondria converts yellow colored insoluble tetrazolium salt to purple colored water-soluble formazan. Sperm also have mitochondria in the midpiece, therefore sperm viability could be evaluated by MTT reduction assay. Several studies have already demonstrated the capability of application of the MTT reduction assay to sperm of several species in Hepes-BSA buffer. Because most liquid semen was diluted in extender like BTS (Beltsville Thawing Solution), Modena or Androhep when it is used or transferred, semen needed another dilution in Hepes-BSA buffer to assess sperm viability. In this study, we evaluated boar sperm viability especially in BTS extended semen and compared the efficiency of this test with eosin-nigrosin staining. We used the fresh BTS extended semen from a local A.I center. Semen sample was diluted to $3.0{\times}10^7$ sperms/ml in BTS. The rates of formazan production were measured in 96-well microtiter plates immediately and 1h after incubation at $17^{\circ}C$ using a spectrophotometer at wave length 560 nm. Simultaneously, split samples of the same semen were tested, using eosin-nigrosin staining to compare the efficiency of the MTT assay of sperm viability in BTS. The correlation between the results of these tests was calculated using Student-t test and ANOVA. The results revealed a strong correlation between the results of MTT reduction rate and the results that were simultaneously determined by eosin-nigrosin staining at 1 h. In conclusion, the MTT reduction test was an effective and simple method to validate sperm viability and it could be used as a simple tool to evaluate sperm viability in the local A.I center and laboratory.

• 한국수정란이식학회지
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• 제27권1호
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• pp.21-28
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• 2012

A study was conducted on four crossbred bulls, used as artificial insemination (AI) sires, to correlate their semen quality with their non return rate (NRR). Semen was collected once a week via an artificial vagina, diluted in egg yolk-citrate and maintained at $+7^{\circ}C$ for three days. It was evaluated for sperm motility, viability, morphology immediately after collection and was examined daily for sperm motility, viability and morphology of acrosome, mid piece and tail for a total of three days. A total of 2016 cows were inseminated by two AI technicians. The proportions of sperm with normal heads were 83.4% (63.7~91.7%), the proportion of spermatozoa exhibiting normal morphology (acrosome, mid piece and tail), motility and viability were 89.2% (82.3~92.0%), 71.3% (61.7~75.0%) and 76.7% (65.7~85.0%), respectively in fresh ejaculates. Sperm motility and sperm viability was significantly ($p$ <0.05) lower in Holstein-Friesian ${\times}$ Local bull than in other bulls during all three days of storage. The overall NRR for four bulls was 82.7% (72.9-87.5%). Bulls with higher sperm motility, viability and normal morphology of spermatozoa of individual bull had significantly (each $p$ <0.05) higher NRR. The highest ($p$ <0.01) NRR (87.5%) was observed in a Red Chittagong bull whose semen qualities were significantly ($p$ <0.05) higher than Holstein-Friesian ${\times}$ Local bull (NNR 72.9%). The results of the present study concluded that NRR at 56 days post AI is related to parameters of semen quality. Therefore, semen evaluation may allow the discarding of bulls with poor fertility in an AI program.

• Journal of Animal Science and Technology
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• 제48권1호
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• pp.15-20
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• 2006

국내에서 사용한 젖소 정액을 네 가지(한국형 보증종모우 정액, 농협 수입 보증종모우 정액, 미국산 수입 정액, 카나다산 수입 정액)로 분류하여 표현형 능력 및 유전능력을 비교하였다. 유전능력의 경우 농촌진흥청 축산연구소에 수행하고 있는 국가단위 젖소유전능력평가 결과를 이용하였다. 유량, 유지량 및 유단백량의 표현형 능력의 경우 미국산 수입 정액이 가장 우수하였고, 한국형 보증종모우 정액은 미국산 수입 정액과 유의한 차이를 보였다. 1991년 이후 태어난 보증종모우의 경우 유지량은 네 종류의 정액 사이에 유의한 차이가 없었다. 유량, 유지량, 유단백량, 유단백율 유전능력의 경우 네 종류 사이에 유의한 차이가 없었으며, 유지율의 경우 한국형 보증종모우 정액이 가장 우수하였다. 선형심사 최종점수 및 유방종합지수의 경우 카나다산 수입정액이 가장 우수하였으며, 한국형 보증종모우 정액과 유의한 차이를 보였다. MFP 및 KTPI의 경우 네 종류 사이에 유의한 차이가 없었다. 1991년 이후에 태어난 보증종모우의 유량 및 유단백량의 경우 네 종류 사이에 유의한 차이가 없었으나, 유지량, 유지율 및 유단백율의 경우 한국형 보증종모우 정액이 가장 우수하였고, 나머지 정액과 유의한 차이를 보였다. 선형심사 최종점수의 경우 한국형 보증종모우 정액, 미국산 수입정액 및 카나다산 수입정액 사이에 통계적으로 유의한 차이가 없었다. 유방종합지수의 경우 카나다산 수입정액이 가장 우수하였고, 한국형 보증종모우 정액과 유의한 차이가 났다. MFP의 경우 한국형 보증종모우 정액이 가장 우수하였고, 나머지 정액과 유의한 차이를 보였다. KTPI의 경우 한국형 보증종모우 정액, 미국산 수입정액 및 카나다산 수입정액 사이에 통계적으로 유의한 차이가 없었다.

• Reproductive and Developmental Biology
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• 제31권2호
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• pp.121-126
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• 2007

The objective of this study was to establish the optimal conditions for hypoosmotic swelling (HOS) test to assess the functional integrity of the membranes of boar fresh or frozen/thawed spermatozoa. When pooled semen sample was incubated for 30 min at $37^{\circ}C$ with different test solution of varied osmolarity, the highest percentage of HOS positive spermatozoa was observed in a 150 mOsmol fructose/Na-citrate solution (33.6%). Incubation time did not affect significantly the score of HOS positive spermatozoa observed in a 150 mOsmol fructose/Na-citrate solution at $37^{\circ}C$, but the osmolarity affected the score of HOS positive spermatozoa under the same condition above. Fresh semen was significantly better than frozen/thawed semen in semen parameters evaluated such as motility, viability, membrane integrity and lipid peroxidation (p<005). In the relationships of sperm parameters, motility vs viability, motility vs membrane integrity and viability vs membrane integrity were positively correlated ($0.82{\sim}0.94$) but lipid peroxidation vs other estimated factors was negatively correlated ($- 0.90{\sim}- 0.98$). Among the evaluation methods, motility vs Viability, motility vs membrane integrity and lipid peroxidation vs other estimated factors were significantly correlated (p<0.05). These results of this. study indicate that the optimal condition of HOST in boar spermatozoa is a 150 mOsmol fructose/Na-citrate solution for 30 min incubation at $37^{\circ}C$ and HOST can substitute the examination of motility, viability and lipid peroxidation.

• Clinical and Experimental Reproductive Medicine
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• 제42권3호
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• pp.101-105
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• 2015

Objective: Several publications have established a relationship between sperm DNA damage and male factor infertility, based on data from America, Europe, and Asia. This study aimed to compare the extent of sperm DNA damage in sperm samples from Nigerian men with unexplained infertility and in sperm samples from a fertile group composed of sperm donors who had successfully impregnated a female partner naturally or through assisted conception. Methods: A total of 404 men underwent male fertility evaluation at Androcare Laboratories and Cryobank participated in this study. Semen analysis and a sperm chromatin structure assay (SCSA) were performed on all subjects. Results: The men in the unexplained infertility group were slightly older than the men in the fertile sperm group ($36{\pm}10$ years vs. $32{\pm}6$ years, p=0.051). No significant difference was observed between the two groups in semen analysis parameters ($p{\geq}0.05$). Men in the unexplained infertility group with normal semen parameters had a significantly higher DNA fragmentation index (DFI) than men in the fertile sperm group ($27.5%{\pm}7.0%$ vs. $14.1%{\pm}5.3%$, p<0.05). In the unexplained infertility group, 63% of the men had a DFI greater than 20%, compared to 4% in the fertile sperm group. In the unexplained infertility group, 15.2% of the subjects had a DFI greater than 30%, compared to 1% in the fertile sperm group. Conclusion: Our study showed that the SCSA may be a more reliable predictor of fertility potential than traditional semen analysis in cases of unexplained infertility.

• Asian-Australasian Journal of Animal Sciences
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• 제23권10호
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• pp.1276-1281
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• 2010

The present study was conducted to investigate the effects of butylated hydroxytoluene (BHT) supplementation on diluted, cooled and frozen-thawed ram spermatozoa. After primary evaluation of collected ejaculates, only semen samples with motility of more than 70% and sperm concentration higher than $3{\times}10^3$ sperm/ml were used for cryopreservation. The selected semen samples were then pooled and diluted 1:4 with Tris Citrate Fructose Yolk (TCFY) extender supplemented with different concentrations of BHT (0.5, 10, 2.0 and 3.0 mM). As the control, semen was diluted and frozen in the diluent without BHT. Motility, progressive motility, viability, membranes and acrosome integrity were evaluated after dilution (part 1), cooling (part 2) and freezing and thawing (part 3). The results of the first part of the experiment showed that there were no significant difference between treatments in the motility, progressive motility, viability, membranes and acrosome integrity of spermatozoa, but the results with 2.0 mM BHT were slightly better than obtained with other levels of BHT and control extender. Significantly better results (p<0.05) were observed in the second part of the experiment for cooled spermatozoa characteristics, when extender was supplemented with 2.0 and 3.0 mM BHT. Furthermore, the results obtained in the third part of the experiment indicated that, after freezing and thawing, all evaluated semen characteristics were improved significantly (p<0.05) by increasing BHT levels, with the best results obtained for extender containing 2 mM BHT. Comparison of these results with those of control diluent, the effects of supplementation were significantly (p<0.01) better. However, the higher concentration of BHT (3.0 mM) reduced the motility, acrosomal integrity, viability and hypo-osmotic swelling response of spermatozoa compared to extender containing 2.0 mM BHT. In conclusion, the results obtained in this study showed that the semen quality of rams was improved when BHT was added to extender used before the freezing process.

• 한국축산식품학회지
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• 제37권4호
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• pp.502-510
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• 2017

This study investigated bacterial growth-inhibitory effect of 69 therapeutic herbal plants extracts on 9 bacterial strains using a disc diffusion assay. Especially, the antimicrobial activity of Psoraleae semen, which showed different activity on pathogenic Gram-positive and Gram-negative bacteria, was evaluated by MIC (minimal inhibition concentration) and biofilm formation assay. The effect of Psoraleae semen extract on bacterial cell membranes was examined by measurement of protein leakage (optical density at 280 nm) and scanning electron microscope (SEM). No clear zone was formed on discs containing Gram-negative bacteria, but Gram-positive bacteria exhibited clear zones. The MICs of Psoraleae semen extract were $8{\mu}g/mL$ for Streptococcus mutans, and $16{\mu}g/mL$ for Enterococci and Staphylococcus aureus. In addition, biofilm formation was inhibited at concentration $8-16{\mu}g/mL$. Protein leakage values and SEM images revealed that cell membranes of Gram-positive bacteria were impaired following exposure to the extract. Further, the extract inhibited the growth of Listeria monocytogenes in sausages. These results indicate that Psoraleae semen extract could be utilized as a natural antimicrobial agent against Gram-positive bacteria.