백개자, 나복자 및 두 배합 약물의 천식 동물 모델에 대한 효과

김창민 ( Chang Min Kim ),이영철 ( Young Cheol Lee ),이장천 ( Jang Cheon Lee ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.6

Objectives: To clarify the possible effects of Sinapis Semen and Raphani Semen on the development of pulmonary eosinophilic inflammation in a asthmatic mouse model. Methods : BALBav/c mice were sensitized to OVA followed intratracheally and by aerosol allergene challenges. We investigated the effect of Sinapis Semen and Raphani Semen on airway hyperresponsiveness, eosinophiic infitratio, immune cell phenotype, The2 cytokine product, and OVA-spedific IgE production. Results : Total lung cells, eosinophils, and lung leukocytes, OVA specific IgE levels, and Th 2cytokine levels such as IL-5, IL-13, IL-17, TNF-alpha, and eotaxin in BALF were reduced compared with those of OVA sensitized asthma mice (control). The absolute numbers of CD3+, CD3+/CD69+, CD3-/CCR3+, CD4+, CD8+, Gr-1+/CD11b+, B220+/CD22+, B220+/IgE+ cells in lung tissiues significantly reduced compared to those of control. Specially total lung cells in BALF and the absolute number of CD3+/CD69+ and, B220+/IgE+ cells in lung tissiue effectively reduced in Sinapis Semen plus Raphani Semen compared to those of Sinapis Semen and Raphani Semen. Conclusions : These results indicate that Sinapis Semen plus Raphani Semen has deep inhibitory effects on airway inflammation and hyperresponsiveness in asmatic mouse model and also has effect of suppression of IL-5, IL-13, IL-17, OVA specific IgE production in BALF. The results verified that Sinapis Semen, Raphani Semen, and Sinapis Semen plus Raphani Semen could act as a immunomodulator which possess anti-inflammatory and anti-asthmatic property by modulating the relationship of Th1/Th2 cytokine imbalance.

A Study on the Characterization of Boar Semen and the Infectious Aspects of Pathogenic Viruses

Yong-Sang Park,Min-Hee KO,Moon-Suck Ko,Tae-Yong Kang 한국동물생명공학회(구 한국동물번식학회) 2011 발생공학 국제심포지엄 및 학술대회 Vol.2011 No.1

Sperm examination is an important tool in estimating the fertilizing capacity of an ejaculate. The number of spermatozoa in a semen dose, morphology and motility are important for the fertilization process. By evaluation of semen, artificial insemination (AI) using high quality of semen can increase fertilization rate. Boar semen is subject to contamination by various pathogens that can result in fertility disorders in sows. Among these pathogens, porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), porcine circovirus-2 (PCV-2) are of particular importance and accurate monitoring prior to and during the presence of boars in AI stations is essential. Because of the high risk of dissemination of disease via AI, The absolute goal is to provide pathogen-free semen and this is feasible with the adequate measures. The disease affects boars semen causes a significant reduction quality. In this study we investigated the characterization boar semen in Jeju, interaction of pathogenic virus infection with characterization of boar semen. Forty-two boar semen from 13 farms were investigated. The semen were stored during 5 days at 17℃ and the sperm qualities in the stored semen were analysed. Visual-motility assessment is a tool (Computer- Assisted Semen Analysis) used to determine the quality of boar semen. Percentage of morphologically normal spermatozoa were assessed. PRRS ,PPV and PCV-2 were detected in boar semen using PCR. The motion characteristics in boar semen was showed 68.4±9.1% for motility, 48.6±7.1 μm/s for VAP, 45.3±7.0 μm/s for VSL, 79.1±8.7 μm/s for VCL, 1.3±0.2 μm/s for ALH, 8.3±0.4 Hz for BCF, 93.6±3.5% for STR, 57.9±6.4 % for LIN. The percentage of sperm with abnormal head, midepeice and tail were 0.3±0.7%, 14.4±12.5%, 4.9±6.6%, respectively. Based on the PCR method, PPV was detected in 20 samples (48%). However, PCV-2 and PRRSV were not detected in any cases. Marked differences in motility and morphology between PPV negative and PPV positive semen were not observed. Sperm cell production was not affected by PPV infection. However, slight increases in detached head, coiled tail after infection were observed (p<0.05). The motility of semen in Jeju is similar to case comparing with other regions in Korea. Although PPV in semen was not affected in semen quality, there is the high risk of virus excretion in the semen of Jeju boars. Therefore continuous screening tests for some particular pathogens in boar semen would be warranted.

The effects of TRIS and TES based semen diluents on the viability of frozen dog spermatozoa

Sung Woo Kim,Si Eun Kim,Jae-Yeong Lee,Chan-Lan Kim,Yeong Gyu Ko,Sung Soo Lee 한국실험동물학회 2021 한국실험동물학회 학술발표대회 논문집 Vol.2021 No.7

The quality of frozen dog semen has been lower than those of livestock like bulls because the semen diluents were not optimized for separate species. The lowered number of spermatozoa in the canine semen has been bothered the new research on the cryobiology of dog semen. The various semen diluents have been reported to increase the viability of frozen semen and also developed with separate animal species. The dog semen is a necessary genetic material for experimental animals, but the dog semen banking system is still insufficient in Korea. In this study, we developed a simple medium for dog semen freezing. The named RDA-dog semen (RDS) diluent was prepared with TRIS 165.1 mM, TES 43.94 mM, Citric acid monohydrate 51.48 mM, Glucose 36.59 mM. Streptomycin sulfate 0.1114 g and Benzylpenicillin Sodium salt 0.0479g were supplemented in 100ml RDS diluents containing 20% egg yolk to inhibit contamination. The dog semen was collected from the Sapsal dog with globed hand massage methods. The effective utilization of RDS diluent was compared with the motility of frozen spermatozoa was compared with two different dog semen diluent, Tris/Citrate/Glucose diluent and BES-lactose diluent. The fresh dog semen with at least 90% motility from 4 different male dogs were pooled for experiments. The final concentration of sperm was adjusted 80ⅹ106/ml after semen plasma removal with centrifugation. The first diluent without glycerol was used for centrifugation and cooled to 17°C for 1 h and added with the same volume of 8% glycerol containing diluents. After cooling the semen to 5°C for 5~6 h, the semen in 0.5ml straws was exposed to the vaper of LN2 at a 5 cm distance from the surface. After thawing at 37°C for 40 sec, the motility of spermatozoa was significantly different with 53.5±5.5% (Tris/Citrate/Glucose diluent), 38.4±5.3% (BES-lactose diluent) and 68.3±3.6% (RDS diluent) (p<0.05). These results indicated that lactose-based diluent is not the choice of dog breeder, and TRIS and TES concentration is essential for a successful dog semen banking system.

개 분획채취 동결정액의 생존성과 인공수정에 의한 임신율에 관한 연구

김영희,김상근 충남대학교 수의과대학 동물의과학연구소 2004 動物醫科學硏究誌 Vol.12 No.-

This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, removed seminal plasma by removed seminal plasma with saline and tris buffer semen and fractional semen of dogs, and the effect of temperature and preservation time and cryoproservation on motility of whole and removed seminal plasma with salin and tris buffer semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. The volume per ejaculate semen, sperm of concentration and motility and abnormal sperm rate of lst fractional semen were 0.65±0.09ml, 4.52±0.35×10^(6)cells/ml, 15,64±3.85% and 5.50±0.62%. Also, 2nd fractional semen were 1.25±0.20ml, 3.35±0.48×10^(6)cells/ml, 96.25±4.65% and 4.24-0.46%. And 3rd fractional semen were 1.45±0.21ml, 3.85±0.52×10^(6)cells/ml, 92.82±4.24% and 4.66±0.58%, respectively. The sperm of concentration and motility and abnormal sperm rates of whole, removed seminal plasma with saline and tris buffer semen were 5.45±0.82×10^(6) cells/ml. 95.55±4.65%, 4.58-0.45% and 4.82±0.36×10^(6)cells/ml, 90.10± 3.42%, 6.48±0.68% and 4.55±0.45×10^(6)ce11s/ml, 93.25±3.85%, 4.82±0.58%, respectively. The motility of whole, removed seminal plasma with saline and tris buffer semen were higher at 4℃ than at 38℃. When preservation temperature was at 4℃, survival rates of removed seminal plasma with saline and tris buffer sperm were 97.54%~6.25% at 1~72 hrs, 97.40%~5.62% at 1~100 hrs, respectively. The survival rates of slow and rapid frozen 2nd fraction, removed seminal plasma with salin and tris buffer semen were 67.3±4.45%, 88.8±4.46% and 46.4±3.84%, 74.4±4.20%, respectively. Survival rates was significantly higher in frozen removed seminal plasma with saline and tris buffer semen than that in control group (8.5±2.12%).

구자, 총실 및 양총실의 형태 감별에 관한 연구

진주환 ( Ju Hwan Jin ),길기정 ( Ki Jung Kil ),이영종 ( Young Jong Lee ) 대한본초학회 2004 大韓本草學會誌 Vol.19 No.2

N/A Objective : Allii Tuberosi Semen and Allii Fistulosi Semen have been sold mixed with fabricated forms of them in Korea. hn such a situation, seeds of Allium tuberosum Rottler, A. fistulosum Linne, and A. cepa Linne were collected and their external, internal, and flour states were observed through microscopic examination. Method : The slice of the tested material made by paraffin section technique was colored with Safranine Malachite Green contrast methods, and the flour of it was mounted by the liquid made by the same ratio of each of glycerin, acetic acid, and water, and then observed and photographed by olymphus-BHT. Result 1. Allii Tuberosi Semen showed the salient network-like wrinkles on its upper side, but Allii Fistulosi Semen and Allii Cepae Semen had 1 -2 lines on their upper sides. 2. The length of Allii Cepae Semen was 2.5?3.5mm, and the width of it was 1.5--2.5mm, which was a bit small; but the length of Allii Fistulosi Semen was 3-4mm, and the width of it was 2?3mm, which was a bit large. 3. In the flour forms, Allii Tuberosi Semen has network-like pattern in the surface of the seed coat epidermal cell wall, but Allii Fistulosi Semen and Allii Cepae Semen have litte or no such pattern. Conclusion : Because there are some clear differences in external and flour states of Allii Tuberosi Semen. Allii Fistulosi Semen, and Allii Cepae Semen, microscopic examination can be used to distinghish them and their fake forms in flour states.

Antibiotic Resistant Microbial Contamination (Enterobacter cloacae) Derived from Egg Yolk and Frozen Semen Extender in Porcine In Vitro Fertilized Embryos

곽성성,Se-Heon Jeong,Seung-Hoon Jang,Yubyeol Jeon,Young-Hee Nam,Dibyendu Biswas,Wan-Kyu Lee,현상환 사단법인 한국동물생명공학회 2010 한국동물생명공학회지 Vol.25 No.4

The present study was to investigate the source of contamination during semen processing for in vitro uses. In the present study, frozen semen was prepared from liquid semen in our laboratory for in vitro fertilization (IVF) experiments due to lack of fresh semen. Antibiotics were added in the frozen semen extender (kanamycin and gentamicin)and in vitro culture (IVC) medium (gentamicin) for further inhibiting growth of microorganisms. Nevertheless, proliferations of microorganisms were observed in IVC culture drop during culturing of IVF embryos using frozen semen. Randomly 3 samples were taken from the liquid semen, frozen semen and egg yolk. Contaminated IVC medium,frozen-thawed semen, liquid semen and egg yolk were cultured in de Man, Rogosa and Sharpe (MRS) agar medium. Whitish colonies were detected in contaminated IVC drop, frozen-thawed semen samples and egg yolk but no colonies were formed in liquid semen samples. Gram-negative and rod-shaped identical bacteria were found in both frozen-thawed semen sample and contaminated IVC drop and egg yolk samples. Enterobacter cloacae were confirmed by API 20E kit according to manufacturer’s instruction with identification value (% ID) 94.3% and T index 0.88. Antibiotic susceptibility tests were done according to Clinical and Laboratory Standards Institute (CLSI) by using ampicillin, amikacin,cephalothin, gentamicin, kanamycin, tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin test. Among them Enterobacter cloacae were resistant to ampicillin, amikacin, cephalothin, gentamicin,kanamycin but susceptible to tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin. From these findings it could be suggested that this contamination sources might be from egg yolk.

Stereoscope를 이용한 미세종자류 한약재 외부형태 감별연구(제2보(報)) -구자(?子)와 총자(蔥子), 동규자(冬葵子)와 경마자(?麻子), 차전자(車前子)와 형개자(荊芥子)-

김영식,주영승,Kim, Young-Sik,Ju, Young-Sung 대한본초학회 2016 大韓本草學會誌 Vol.31 No.4

Objectives : This study is to propose discriminative keys with stereoscope for minute seed herbs easily mixed up, but hard to discriminate by naked eyes: Alli Tuberosi Semen(ATS)?Alli Fistulosi Semen(AFS), Malvae Semen(MS)? Abutili Semen(AS), Plantaginis Semen(PS)?Schizonepetae Semen(SS).Objectives : We reviewed the description of original plants and medicinal herbs from literature. The specimen of original plant were collected, determinated and compared to samples in the market. Primary classification was performed with naked eyes. and we found out other discrimination keys for non-distinctive herbs with stereoscope. Discrimination keys were set as the morphological criteria of authentic herbs, percentage of adulteration, and standards for discriminating genuine herbs from adulteration.Results : 1) ATS is bigger, has reticulate pattern on protuberant side and unique garlic chives taste. AFS which is usually mixed up, is smaller, has 1~2 ridges and unique welsh onion taste. 2) MS is smaller, has no villus, but its length of bumps are similar. AS easily mixed up, is bigger and has white villus. Its upper side bump is longer and more sharp. 3) PS has dent at middle of the back side. It becomes very sticky when it is put or boiled in water. SS which can be mixed up has no dent and low viscosity compared to PS.Conclusions : With this result, we propose discriminative keys which can identify the original plants and processed herbal state of six herbs. Because minute medicinal herbs are hardly distinguishable by sensory test, It is essential to differentiate by using stereoscope.

Characterization and cryopreservation of Amur leopard cats (<i>Prionailurus bengalensis euptilurus</i>) semen collected by urethral catheterization

Jeong, Dong-Hyuk,Kim, Jeong-Ho,Na, Ki-Jeong Elsevier 2018 Theriogenology Vol.119 No.-

<P><B>Abstract</B></P> <P>The Amur leopard cat (<I>Prionailurus bengalensis euptilurus</I>) is a globally endangered species, and there is thus an urgent need to increase its population. The objectives of this study were to: (1) confirm the utility of urethral catheterization (UC) for semen collection from Amur leopard cats; (2) investigate proper dilution media for fresh semen; and (3) investigate the effectiveness of sperm cryopreservation, including examining the effect of glycerol concentration. Six adult males in captivity (mean weight 5.03 ± 0.44 kg, aged 2–6 years) were included. All study procedures were performed during the breeding season (February to April) over two consecutive years. Semen samples (n = 28) were collected four or five times from each animal (four times for two animals and five times for four animals) by UC under general anaesthesia, and their characteristics (including sperm motility) were evaluated. First, the sperm motility in semen diluted in Ham's F-10 or phosphate buffer saline (PBS) was compared. Next, semen diluted with TEST-yolk buffer containing 2%, 4%, or 6% glycerol was frozen in a liquid nitrogen tank, and sperm motility and acrosome integrity were evaluated after thawing. No difference in motility was observed between sperm diluted in Ham's F-10 and PBS. The percentages of sperm motility and kinetic values in semen frozen in 2% or 4% glycerol were higher than those in semen stored in 6% glycerol. In conclusion, the UC method for semen collection is recommendable for Amur leopard cats and should be useful for artificial insemination. Although sperm motility decreased after thawing, samples thus preserved may be usable for advanced reproductive techniques, such as intracytoplasmic sperm injection or in-vitro fertilization.</P> <P><B>Highlights</B></P> <P> <UL> <LI> This is the first study on semen collection by urethral catheterization (UC) in Amur leopard cats (ALC). </LI> <LI> We confirmed multiple benefits of UC for semen collection in ALC. </LI> <LI> There was no difference of semen quality in Ham’s F-10 or phosphate buffer saline for 3 h during cool storage. </LI> <LI> Better results were obtained with a TEST-Yolk buffer with glycerol 2% and 4% than 6% after freezing of the semen. </LI> <LI> Thawed semen may be used for advanced reproductive techniques. </LI> </UL> </P>

Stereoscope를 이용한 미세종자류 한약재 외부형태 감별연구 제2보(제2報) -구자(구子)와 총자(蔥子), 동규자(冬葵子)와 경마자(경麻子), 차전자(車前子)와 형개자(荊芥子)-

김영식 ( Young-sik Kim ),주영승 ( Young-sung Ju ) 대한본초학회 2016 大韓本草學會誌 Vol.31 No.4

Objectives: This study is to propose discriminative keys with stereoscope for minute seed herbs easily mixed up, but hard to discriminate by naked eyes: Alli Tuberosi Semen(ATS)*Alli Fistulosi Semen(AFS), Malvae Semen(MS). Abutili Semen(AS), Plantaginis Semen(PS)*Schizonepetae Semen(SS). Methods : We reviewed the description of original plants and medicinal herbs from literature. The specimen of original plant were collected, determinated and compared to samples in the market. Primary classification was performed with naked eyes. and we found out other discrimination keys for non-distinctive herbs with stereoscope. Discrimination keys were set as the morphological criteria of authentic herbs, percentage of adulteration, and standards for discriminating genuine herbs from adulteration. Results : 1) ATS is bigger, has reticulate pattern on protuberant side and unique garlic chives taste. AFS which is usually mixed up, is smaller, has 1~2 ridges and unique welsh onion taste. 2) MS is smaller, has no villus, but its length of bumps are similar. AS easily mixed up, is bigger and has white villus. Its upper side bump is longer and more sharp. 3) PS has dent at middle of the back side. It becomes very sticky when it is put or boiled in water. SS which can be mixed up has no dent and low viscosity compared to PS. Conclusions : With this result, we propose discriminative keys which can identify the original plants and processed herbal state of six herbs. Because minute medicinal herbs are hardly distinguishable by sensory test, It is essential to differentiate by using stereoscope.

Analysis of Semen Parameters in α1,3-Galactosyltransferase-/- Boars

황인설,이승찬,김성우,권대진,박미령,양현,오건봉,옥선아,우재석,임기선,황성수 사단법인 한국동물생명공학회 2017 한국동물생명공학회지 Vol.32 No.2

It is very difficult to get the information about semen quality analysis in transgenic pigs because of limited numbers and research facilities. Therefore, in the present study, we analyzed the semen quality of transgenic boars generated for xenotransplantation research. Briefly, the semen samples were collected from 5 homozygous α1,3-Galactosyltransferase knock-out (GalT-/-) transgenic boars and immediately transported to the laboratory. These semen samples were decupled with DPBS and conducted to analyze semen parameters by a computer-assisted semen analysis (CASA) system. The boar semen were examined all 12 parameters such as total motility (TM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), and hyperactivated (HYP), etc. In results, among the 5 GalT-/- boars, three boars (#134, 144, and 170) showed normal range of semen parameters, but #199 and 171 boars showed abnormal ranges of semen parameters according to standard ranges of semen parameters. Unfortunately, #171 boar showed azoospermia symptom with rare sperm counts in the original semen. Conclusively, assessment of semen parameters by CASA system is useful to pre-screening of reproductively healthy boar prior to natural mating and artificial insemination for multiplication and breeding.