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Detection of Soybean mosaic virus by Reverse Transcription Loop-mediated Isothermal Amplification (Reverse transcription Loop-mediated isothermal amplification을 이용한 Soybean mosaic virus의 진단)

  • Lee, Yeong-Hoon;Bae, Dae-Hyeon;Kim, Bong-Sub;Yoon, Young-Nam;Bae, Soon-Do;Kim, Hyun-Joo;Mainali, Bishwo P.;Park, In-Hee;Lee, Su-Heon;Kang, Hang-Won
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.315-320
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    • 2015
  • Soybean mosaic virus (SMV) is a prevalent pathogen that causes significant yield reduction in soybean production worldwide. SMV belongs to potyvirus and causes typical symptoms such as mild mosaic, mosaic and necrosis. SMV is seed-borne and also transmitted by aphid. Eleven SMV strains, G1 to G7, G5H, G6H, G7H, and G7a were reported in soybean varieties in Korea. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method allowed one-step detection of gene amplification by simple procedure and needed only a simple incubator for isothermal template. This RT-LAMP method allowed direct detection of RNA from virus-infected plants without thermal cycling and gel electrophoresis. In this study, we designed RT-LAMP primers named SML-F3/B3/FIP/BIP from coat protein gene sequence of SMV. After the reaction of RT-LAMP, products were identified by electrophoresis and with the detective fluorescent dye, SYBR Green I under daylight and UV light. Optimal reaction condition was at $58^{\circ}C$ for 60 min and the primers of RT-LAMP showed the specificity for nine SMV strains tested in this study.

Application of Chemiluminescence Enzyme Immunoassay Method to Collect in vivo Matured Oocyte in Dog Cloning (개 복제 시 체내 성숙 난자 회수를 위한 화학발광효소면역분석기법의 적용)

  • Kim, Min-Jung;Oh, Hyun-Ju;Kim, Geon-A;Jo, Young-Kwang;Choi, Jin;Lee, Byeong-Chun
    • Journal of Veterinary Clinics
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    • v.31 no.4
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    • pp.267-271
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    • 2014
  • Accurate determination of in vivo oocyte maturation is particularly critical for dog cloning compared to other assisted reproductive technologies because oocytes in metaphase II stage have to be recovered in order to undergo somatic cell nuclear transfer right after its recovery. The aim of present study was to evaluate the reliability and to set a reference range of a chemiluminescence enzyme immunoassay (CLEIA) compared to radioimmunoassay (RIA) method to retrieve in vivo matured oocytes. Serum progesterone concentration during proestrus and estrus was analyzed by RIA and CLEIA to determine ovulation day (Day 0). On Day 3, in vivo oocytes were recovered surgically and evaluated microscopically maturation status after staining nucleus with bisbenzimidazole dye. Mean progesterone concentration by CLEIA ($7.64{\pm}0.06ng/ml$) was significantly higher than by RIA ($6.46{\pm}0.04ng/ml$, P < 0.0001). It was not different between CLEIA ($10.01{\pm}0.34ng/ml$) and RIA values ($7.91{\pm}0.14ng/ml$, P < 0.05) on Day 0, but significantly higher CLEIA level on Day -1 and Day 1 ($6.41{\pm}0.15$ and $14.25{\pm}0.44ng/ml$) was assessed compared to RIA ($4.95{\pm}0.10$ and $11.29{\pm}0.34ng/ml$). However, with both methods, progesterone level was significantly increased from Day -1 to Day 2. To determine oocyte maturation with CLEIA method, a wider and higher reference range has to be considered.

The Effect of Glide Path on Canal Centering Ability in Reciprocating File System (Reciprocating 파일 시스템에서 Glide Path가 근관만곡도 유지에 미치는 영향)

  • Zang, Ki-Choul;Kim, Jin-Woo;Cho, Kyung-Mo;Park, Se-Hee
    • Journal of Dental Rehabilitation and Applied Science
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    • v.28 no.3
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    • pp.245-252
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    • 2012
  • The purpose of this study was to evaluate the influence of glide path on canal centering ratio after instrumentation with different single file systems; WaveOne and Reciproc. Reciproc R25 (VDW), WaveOne Primary (Dentsply Maillefer) and PathFile #13, 16, 19 (Dentsply Maillefer) were used in this study. In no glide path groups, Reciproc files and WaveOne files used for canal preparation without glide path. In glide path groups, the PathFile were used before canal preparation. Methylene blue dye was introduced into the canal to obtain a clear pre-instrumentation image. Pre-instrumentation images and post-instrumentation images were scanned using Epson Perfection V700 Photo scanner (Epson, Nagano, Japan). Transparencies of post-instrumentation images were changed and superimposed on pre-instrumentation images using Adobe Photoshop CS 3 (Adobe Systems Incorporated, San Jose, CA, USA). The centering ratio was calculated for each instrumented canal using the following formula: CR=|X1-X2|/Y. It was statistically analyzed using two-way ANOVA at 95% confidential level. The centering ratio in glide path groups were significant less than it in no glide path groups at 3, 4, 5 and 6 mm level. Except 1 and 6 mm level, WaveOne groups had significant less centering ration than Reciproc groups. At 6 mm level, there was no significant difference between WaveOne and Reciproc. In the limitation of this study, creation of a previous glide path before reciprocating motion instrumentation in curved canal appears to be appropriate and WaveOne system can be used for preparation of curved canal without severe aberrations.

Anti-obesity effect of Polygala tenuifolia (원지(Polygala tenuifolia)의 항비만 효과)

  • Hwang, Ju-Young;Wu, Yong-Xiang;Hwang, Dae-Il;Bae, Suk-Jae;Kim, Taewan
    • Korean Journal of Food Preservation
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    • v.21 no.1
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    • pp.97-106
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    • 2014
  • This study was performed in order to investigate the anti-obesity effect of Polygala tenuifolia on lipid mechanism in 3T3-L1 adipocytes. The chemical composition of the P. tenuifolia was analyzed in order to assess its nutritional value. Total dietary fiber was the highest among the proximate component of the P. tenuifolia. These results showed that the P. tenuifolia may be used as a potential functional ingredient for anti-obesity effect. Intracellular lipid droplets in the adipocyte were stained with oil-red O dye and quantified. In comparison to the control, lipid accumulation was significantly decreased by 40.1% and 22.4% when treated with the water extract and 70% EtOH extract of the P. tenuifolia at the concentration of $10{\mu}g/mL$, respectively. The anti-adipogenic effect of the water extract was stronger than that of the 70% EtOH extract. The gene expression levels were measured via Western blot and real-time PCR. As a result, the water extract was found to have decrease the gene expression of SREBP-1c, PPAR, $C/EBP{\alpha}$, FAS, ACC in a dose-dependent manner. These indicate that the water extract inhibits pre-adipocyte differentiation and adipogenesis by blocking the SREBP-1c gene expression in 3T3-L1 cells. Therefore, P. tenuifolia can be used as an effective anti-obesity agent.

Analysis of Research on Non-Timber Forest Plants - Based on the Articles Published in the Journal of Korean Forest Society from 1962 to 2013 - (산림과학분야의 산림특용자원식물의 연구 - 한국임학회지에 게재된 논문을 중심으로 -)

  • Lee, Hyunseok;Yi, Jaeseon;An, Chanhoon;Lee, Jeonghoon
    • Journal of Korean Society of Forest Science
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    • v.104 no.3
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    • pp.337-351
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    • 2015
  • The articles, published in the Journal of Korean Forest Society from Volume 1 (1962) to Volume 102 (2013), were investigated for the research trend analysis about forest plants for special purposes, i.e., edible plants, medicinal plants, feed resources, landscape plants, fiber plants, industrial usage, bee plants, bioenergy/phytoremediation uses, dye materials, and rare/endangered/endemic plants. These research articles were classified again based on the contents of research into following categories - habitat environment, ecology, physiology, propagation, silviculture (including planting and tending), genetics and breeding, identification, pest and disease control, animal-related research, components analysis and extracts, vegetation survey, biotechnology, management, recreation and forest healing, and research review. Among the total 2,433 articles published, 611 (25.1%) were related to plants for special usage or purposes. The highest frequency (14.9%) in publications was found in the field of silviculture followed by physiology, propagation, identification, and genetics and breeding, respectively. On the bases of usage, edible plants showed higher frequency (26.5%) than others, followed by industrial purpose, bioenergy/phytoremediation usage, landscape plants, medicinal plants, and rare/endangered/endemic plants. Populus plant species was the most popular in research, showing 62 articles; and Castanea crenata 36; Pinus koraiensis 35; Robinia pseudoacacia 20; Ginko biloba 17, etc. Based on the survey and analysis, the following points are suggested: 1) improved evaluation of forest plants as non-wood resources, 2) expanding research topics on the basis of production, management, and utilization of non-wood forest resources, 3) management of database of forest plant information and encouragement needed to strengthen cooperative researches satisfying the needs of other industrial and scientific areas, and 4) encouraging to promote traditional knowledge based research on forest plants.

Studies of the Effects of Acupuncture Stimulation at Huatuo Jiaji(EX B2) Points on Axonal Regeneration of Injured Sciatic Nerve in the Rats (화타협척혈 침자극에 의한 손상 말초신경의 재생효과에 관한 연구)

  • Kim, Dae-Feel;Park, Young-Hoi;Keum, Dong-Ho
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.4
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    • pp.39-61
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    • 2008
  • Objectives : The present study was performed to investigate whether acupuncture stimulation in the rats affected regeneration properties of the injured sciatic nerve. A differential effect of acupuncture stimulation on the one point near the spinal nerve root controlling sciatic nerve activity and the other point in the peripheral area subordinated by injured nerve was compared. Materials and Methods: Rat sciatic nerves were injured by crush, and the effects on axonal regeneration on injured sciatic nerves were evaluated by acupuncture stimulation at two different regions. In proximal acupuncture stimulation group, acupuncture stimulation was performed on Huatuo Jiaji(EX B2) points located from L5 to S1 vertebral levels to stimulate the nearest spinal nerve root that innervates sciatic nerves. In distal acupuncture stimulation group, acupuncture stimulation was performed on Zusanli(ST 36) and Weizhong(BL 40) points to stimulate at peripheral area dominated by injured sciatic nerves. Acupuncture stimulation was given every other days for 1 or 2 weeks. Sciatic nerve tissues collected from acupuncture stimulation experimental groups, injury control group, and intact animal group were used for protein analysis by Western blotting or Hoechst nuclear staining. To determine axonal regeneration, Dil fluorescence dye was injected into the sciatic nerve 0.5 cm distal to the injury site in individual animal groups and Dil-labeled cells by retrograde tracing were measured in the DRG at lumbar 5 or in the spinal cord. DRG sensory neurons prepared from individual animal groups were used to measure the extent of neurite outgrowth and for immunofluorescence staining with anti-GAP-43 antibody. Results : Animal groups given proximal or distal acupuncture stimulation showed upregulation of GAP-43 and Cdc2 protein levels in the sciatic nerve at 7 days after injury. Cdk2 protein levels were strongly induced by nerve injury, but did not show changes by acupuncture stimulation. Phospho-Erk1/2 protein levels were elevated by acupuncture stimulation above those present in the injury control animals. These increase in regeneration-associated protein levels appeared to be related with increase cell proliferation in the injured sciatic nerves. Hoechst 33258 staining of sciatic nerve tissue to visualize nuclei of individual cells showed increased Schwann cell number in the distal portion of the injured nerve 7 and 14 days after injury and further increases by acupuncture stimulation particularly at the proximal position. Measurement of axonal regeneration by retrograde tracing showed significantly increased Dil-labeled cells in proximal acupuncture stimulation group compared to distal acupuncture stimulation group and injury control group. Finally, an evaluation of axonal regeneration by retrograde tracing showed increased number of Dil labeled cells in the DRG at lumbar 5 or in the ventral horn of the spinal cord at lower thoracic level at 7 days after nerve injury. Conclusions : The present data show that the proximal acupuncture stimulation at Huatuo Jiaji(EX B2) points governing injured sciatic nerves was more effective for axonal regeneration than the distal acupuncture stimulation. Further studies on functional recovery or associated molecular mechanisms should be critical for developing animal models and clinical applications.

THE EFFECT OF IRRADIATION MODES ON POLYMERIZATION AND MICROLEAKAGE OF COMPOSITE RESIN (광조사 방식이 복합레진의 중합과 누출에 미치는 영향)

  • Park, Jong-Jin;Park, Jeong-Won;Park, Sung-Ho;Park, Ju-Myong;Kwon, Tae-Kyung;Kim, Sung-Kyo
    • Restorative Dentistry and Endodontics
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    • v.27 no.2
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    • pp.158-174
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    • 2002
  • The aim of this study was to investigate the effect of light irradiation modes on polymerization shrinkage, degree of cure and microleakage of a composite resin. VIP$^{TM}$ (Bisco Dental Products, Schaumburg, IL, USA) and Optilux 501$^{TM}$ (Demetron/Kerr, Danbury, CT, USA) were used for curing Filtek$^{TM}$ Z-250 (3M Dental Products, St. Paul., MN, USA) composite resin using following irradiation modes: VIP$^{TM}$ (Bisco) 200mW/$\textrm{cm}^2$ (V2), 400mW/$\textrm{cm}^2$ (V4), 600mW/$\textrm{cm}^2$ (V6), Pulse-delay (200 mW/$\textrm{cm}^2$ 3 seconds, 5 minutes wait, 600mW/$\textrm{cm}^2$ 30seconds, VPD) and Optilux 501$^{TM}$ (Demetron/Kerr) C-mode (OC), R-mode (OR). Linear polymerization shrinkage of the composite specimens were measured using Linometer (R&B, Daejeon, Korea) for 90 seconds for V2, V4, V6, OC, OR groups and for up to 363 seconds for VPD group (n=10, each). Degree of conversion was measured using FTIR spectrometer (IFS 120 HR, Bruker Karlsruhe, Germany) at the bottom surface of 2 mm thick composite specimens V2, Y4, V6, OC groups were measured separately at five irradiation times (5, 10, 20, 40, 60 seconds) and OR, VPD groups were measured in the above mentioned irradiation modes (n=5 each). Microhardness was measured using Digital microhardness tester (FM7, Future-Tech Co., Tokyo, Japan) at the top and bottom surfaces of 2mm thick composite specimens after exposure to the same irradiation modes as the test of degree of conversion(n=3, each). For the microleakage test, class V cavities were prepared on the distal surface of the ninety extracted human third molars. The cavities were restored with one of the following irradiation modes : V2/60 seconds, V4/40 seconds, V6/30 seconds, VPD , OC and OR. Microleakage was assessed by dye penetration along enamel and dentin margins of cavities. Mean polymerization shrinkage, mean degree of conversion and mean microhardness values for all groups at each time were analyzed using one-way ANOVA and Duncan's multiple range test, and using chi-square test far microleakage values. The results were as follows : . Polymerization shrinkage was increased with higher light intensity in groups using VIP$^{TM}$ (Bisco) : the highest with 600mW/$\textrm{cm}^2$, followed by Pulse-delay, 400mW/$\textrm{cm}^2$ and 200mW/$\textrm{cm}^2$ groups, The degree of polymerization shrinkage was higher with Continuous mode than with Ramp mode in groups using Optilux 501$^{TM}$ (Demetron/Kerr). . Degree of conversion and microhardness values were higher with higher light intensity. The final degree of conversion was in the range of 44.7 to 54.98% and the final microhardness value in the range of 34.10 to 56.30. . Microleakage was greater in dentin margin than in enamel margin. Higher light intensity showed more microleakage in dentin margin in groups using VIP$^{TM}$ (Bisco). The microleakage was the lowest with Continuous mode in enamel margin and with Ramp mode in dentin margin when Optilux 501$^{TM}$ (Demetron/Kerr) was used.

Ecophysiological Interpretations on the Water Relations Parameters of Trees(VII) - Measurement of Water Flow by the Heat Pulse Method in a Larix leptolepis Stand - (수목(樹木)의 수분특성(水分特性)에 관(關)한 생리(生理)·생태학적(生態學的) 해석(解析)(VII) - Heat pulse법(法)에 의한 낙엽송임분(林分)의 수액류속(樹液流速) 계측(計測) -)

  • Han, Sang Sup;Kim, Sun Hee
    • Journal of Korean Society of Forest Science
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    • v.82 no.2
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    • pp.152-165
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    • 1993
  • This is the basic study in order to know the amount of transpirational water loss in a Larix leptorepis stand by a heat pulse method. Especially this study has been measured and discussed the diurnal and seasonal trends of heat pulse velocity by changes of radiation, temperature and humidity, differences of heat pulse velocity by direction and depth in stem, differences of heat pulse velocity by dominant, codominant and suppressed trees, diurnal change of heat pulse velocity by change of leaf water potential, sap flow path way in sapwood by dye penetration and amount of daily and annual transpiration in a tree and stand. The results obtained as follows : 1. Relation between heat pulse velocity(V) and sap flow rate(SFR) was established as a equation of SFR=1.37V($r=0.96^{**}$). 2. The sap flow rate presented in the order of dominant, codominant and suppressed tree, respectively. The daily heat pulse velocity was changed by radiation, temperature and vapor pressure deficit. 3. The heat pulse velocity in individual trees did not differ in early morning and in late night, but had some differed from 12 to 16 hours when radiation was relatively high. 4. The heat pulse velocity and leaf water potential showed similar diurnal variation. 5. The seasonal variation of heat pulse velocity was highest in August, but lowest in October and similar value of heat pulse velocity in the other months. 6. The heat pulse velocity in stem by direction was highest in eastern, but lowest in southern and similar velocity in western and northern. 7. The difference of heat pulse velocity in according to depths was highest in 2.0cm depth, medium in 1.0cm depth, and lowest in 3.0cm depth from surface of stem. 8. The sap flow path way in stem showed spiral ascent turning right pattern in five sample trees, especially showed little spiral ascent turning right in lower part than 3m hight above ground, but very speedy in higher than 3m hight. 9. The amount of sap flow(SF) was presented as a equation of SF=1.37AV and especially SF in dominant tree was larger than in codominant or suppressed tree. 10. The amount of daily transpiration was 30.8ton/ha/day and its composition ratio was 83% at day and 17% at night. 11. The amount of stand transpiration per month was largest in August(1,194ton/ha/month), lowest in May (386ton/ha/month). The amount of stand transpiration per year was 3,983ton/ha/year.

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Fracture load and marginal fitness of zirconia ceramic coping by design and coloration (유색 및 백색 지르코니아 세라믹 코핑의 코핑 디자인에 따른 파절 하중과 변연 적합성)

  • Shin, Mee-Ran;Kim, Min-Jeong;Oh, Sang-Chun
    • The Journal of Korean Academy of Prosthodontics
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    • v.47 no.4
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    • pp.406-415
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    • 2009
  • Purpose: The purpose of this study was to compare the marginal fitness and fracture load of the zirconia copings according to the design with different thickness and coloration. Material and methods: The evaluation was based on 80 zirconia copings. Zirconia copings were fabricated in design with different thicknesses using CAD/CAM system (Everset, KAVO dental GmbH, Biberach, Germany). The designs of copings were divided into four groups. The first group consisted of copings with uniform thickness of 0.3 mm. The thickness in the second group was 0.3 mm on the buccal surface and 0.6 mm on the lingual surface. The third group consisted of coping with uniform thickness of 0.6 mm. The thickness in the fourth group was 0.6 mm on the buccal surface and 1mm on the lingual surface. Each group consisted of 10 colored and 10 uncolored copings. Half of the copings (40) processed with a milling system according to the specific design were sent to be given a color (A3) through saturation in special dye by a manufacturing company. Just after sintering, the marginal discrepancies of copings were measured on the buccal, lingual, mesial and distal surfaces of metal die, under a Video Microscope System (sv-35, Sometech, Seoul, Korea) at a magnification of $\times$ 100. It was remeasured after the adjusting of the inner surface. Next, all copings were luted to the metal dies using reinforced cement {GC FujiCEM (GC Corp. Tokyo, Japan)} and mounted on the testing jig in a Universal Testing Machine (Instron 4467, Norwood, MA, USA). The results were analyzed statistically using the one-way ANOVA test. Results: The obtained results were as follow: 1. The measured value of marginal discrepancy right after sintering was the greatest in the contraction of the buccal area in all groups, except for group I2. 2. There was no significant difference of marginal fitness among the groups in the colored zirconia group (P<.05). 3. When the marginal fitness among the groups in the uncolored zirconia group was considered, group II2 had the smallest marginal discrepancy. 4. When the colored and uncolored groups with the same design were compared, there was a significant difference between I1 and II1 groups. In group 2, 3, and 4, the uncolored zirconia had the greatest marginal fitness (P<.05). 5. After adjustment of inner surface, there was no significant difference in the marginal fitness in all groups when color and design of the zirconia coping were compared. 6. The fracture load of CAD/CAM zirconia copings showed significant difference in group 1, 2, 3, and 4. I4 and II4 had the strongest fracture load. 7. When groups with different color and same design were compared, all colored groups showed greater fracture load (P>.05), with no significance. Conclusion: There was difference in the marginal fitness according to the design and coloration of zirconia copings right after sintering, but it was decided that the copings may well be used clinically if the inner surface are adjusted. The copings should be thick enough for the reinforcement of fracture strength. But considering the esthetics of the visible surfaces (labial and buccal surface), the thickness of copings may be a little thin, without giving any significant effect on the fracture strength. This type of design may be considered when giving priority to preservation of tooth or esthetics.

The Effect of Seminal Plasma on Chilling and Freezing of Canine Spermatozoa (개 정액의 정장이 개정자의 냉각과 동결에 미치는 영향)

  • You, Myung-Jo;Lee, John-Hwa;Kim, In-Shik;Park, Jin-Ho;Kwon, Jung-Kee;Kim, Jong-Hoon;Kim, Bum-Seok;Yu, Il-Jeoung
    • Journal of Veterinary Clinics
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    • v.24 no.4
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    • pp.486-492
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    • 2007
  • Seminal plasma(SP) is usually removed from semen that is to be cryopreserved. However, some reports indicate that SP has beneficial effects on spermatozoa during chilling and freezing. The purpose of this study was to determine the effect of SP on sperm survival by adding SP to the extender before cooling and freezing canine spermatozoa. In replicate experiments, ejaculates obtained from four healthy dogs(1-4 years old) of various breeds were pooled, centrifuged at $300{\times}g$ for 10 min at $25^{\circ}C$, and the supernatant of seminal plasma was decanted. Spermatozoa were suspended in egg yolk-Tris(EYT) buffer. The study comprised two experiments: [Exp 1] Sperm were suspended in EYT extender containing either 0, 20, 40, 80 or 100% SP and were slowly cooled to $4^{\circ}C$ for 2h or held at $25^{\circ}C$ as controls. Sperm concentration was adjusted to $2{\times}10^8/ml$. [Exp II] Sperm samples, each of which contained $1{\times}10^8/ml$, were assigned to nine groups to be frozen. In the first four groups, sperm in EYT containing either 20, 40, 80 or 100% SP were cooled to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol and then were frozen. The final concentrations of SP were 10, 20, 40 or 50%. In the other four groups, sperm in EYT alone were first cooled slowly to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol plus 10, 20, 40 or 50% SP and then were frozen. Spermatozoa, which chilled in EYT alone and diluted to contain final concentrations of EYT+0.6M glycerol without seminal plasma, and then frozen, was regarded as control. Spermatozoa were frozen at $25^{\circ}C/min$ of cooling rate in plastic straws that were suspended above liquid nitrogen and thawed in water at $38^{\circ}C$ for 1 min. Sperm survival was assayed by determining progressive motility and integrity of plasma and acrosome membranes. Progressive motility was determined by microscopic examination at $200{\times}$ magnification. Membrane integrity was assessed by use of a double fluorescent dye, and acrosome integrity by staining sperm with Pisum sativum agglutinin. The results of the first experiment showed that adding SP did not improve motility of spermatozoa compared to those incubated without SP regardless of temperature. The results of the second experiment showed that spermatozoa suspended in EYT+0.6M glycerol containing SP exhibited the higher progressive motility before being frozen(P<0.05). However, frozen-thawed spermatozoa that had suspended in EYT+0.6M glycerol containing SP showed the similar or lower viability(P<0.05). In summary, although seminal plasma did not affect spermatozoa that were chilled in EYT without cryoprotectant(CPA), addition of seminal plasma to EYT containing CPA did significantly improved progressive motility of canine spermatozoa that were chilled.