• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Effects of long term storage of semen in liquid nitrogen on the viability, motility and abnormality of frozen thawed Frisian Holstein bull spermatozoa

    2015-12-22 12:09:34AbdulMALIKMuhammadLAILYMuhammadIrwanZAKIR
    Asian Pacific Journal of Reproduction 2015年1期

    Abdul MALIK, Muhammad LAILY, Muhammad Irwan ZAKIR

    1Department of Animal Science, Faculty of Agriculture. Islamic University of Kalimantan, Banjarmasin, Indonesia

    2Office Animal husbandry. District of Tanah Laut, Province South Kalimantan, Indonesia

    Effects of long term storage of semen in liquid nitrogen on the viability, motility and abnormality of frozen thawed Frisian Holstein bull spermatozoa

    Abdul MALIK1*, Muhammad LAILY2, Muhammad Irwan ZAKIR1

    1Department of Animal Science, Faculty of Agriculture. Islamic University of Kalimantan, Banjarmasin, Indonesia

    2Office Animal husbandry. District of Tanah Laut, Province South Kalimantan, Indonesia

    ARTICLE INFO

    Article history:

    Received 16 October 2014

    Received in revised form 18 Novermber 2014

    Accepted 25 November 2014

    Available online 20 March 2015

    Spermatozoa

    Cryopreservation

    Storage

    liquid nitrogen

    Sperm quality

    Objective: To evaluate effects of long term storage of semen in liquid nitrogen on the motility, concentration, viability, and abnormality of frozen-thawed. Methods: A total of four Friesian Holstein bulls were used for this study. One hundred forty semen straws with produced during period from 2008 to 2013 and stored in the liquid nitrogen at the AI center were used in the research. The sample straw was divided into six groups; each group consist 20 semen straws. For group one all straw semen was produced on the 2013 with storage in liquid nitrogen as long as one year, the group 2, 3, 4, 5, and 6 were produced on the 2012, 2011, 2010, 2009 and 2008 with storage in liquid nitrogen as long as 2, 3, 4, 5, and 6 year, respectively. Results: The viability of thawed sperm was not significantly different decreased (P>0.05) between storage on the 1 year and storage on the 2 years. Whereas, the viability was significantly different (P<0.05) with storage on the 3, 4, 5 and 6 years. The motility of thawed sperm was not significantly different decreased (P>0.05) on the storage 1, 2 and 3 years. Whereas, the motility was significantly different (P<0.05) with storage on the 4, 5 and 6 years. The abnormality of thawed sperm was not significantly different increased (P>0.05) on the storage 1, 2 and 3 years. Whereas, the abnormality was significantly increased (P<0.05) with storage on the 4, 5 and 6 years. On the other hand the concentration of thawed sperm was not significantly different decreased (P>0.05) during storage in liquid nitrogen as long as six years. Conclusions: Based on the results in these experiments, it may be concluded that concentration sperm during one year storage in liquid nitrogen resulted in similar concentration storage as long as six years. However, the viability and motility sperm thawed storage in liquid nitrogen during six years was lower than storage on the 1 and 2 years.

    1. Introduction

    Dairy cows in Indonesia can be found in 19 of the 33 provinces. However, ninety seven percent of all dairy cows are located on the Java Island in the three provinces including east Java (47 %), central Java (25 %) and west Java (25 %). One of strategy for development Indonesian’s dairy industry is increasing the populations and productivity of dairy livestock. Artificial insemination (AI) is the good available tool for genetic improvement of dairy cattle in the minimum possible time. One of the factors for success AI is semen quality after thawed as well as relation to biophysical and biochemical characteristics of sperm[1]. Furthermore, Hayashi and Ishobe[2] reported that semen quality frozen thawed including viability, motility, and abnormality is crucial factor for successful AI. High viability, motility and abnormality of spermatozoa frozen thawed are significant factor because the relationship between the post-thawing sperm viability and the subsequent conception rate has been reported[3, 4].

    Cryopreservation in -196 ℃ (liquid nitrogen) is a method that makes long-term storage of spermatozoa[5]. However, for the success of that, it is necessary that spermatozoaare preserved for long periods without damaging their fertilizing ability. Hammerstedt et al[6] and Watson[7] revealed that storage period per time is not having an effect on the sperm viability. However, researches designed to identify a decrease on performance of cryopreserved semen as a function of storage time are deficient. In several cases, the assumption of no damage to spermatozoa during cryopreservation is made subject to the semen being stored unaffected in liquid nitrogen[5].

    The objective of this study was to evaluate effects of long term storage of semen in liquid nitrogen on the motility, concentration, viability and abnormality of frozen-thawed Frisian Holstein bull spermatozoa.

    2. Materials and methods

    2.1. Collection of semen

    This study was conducted at the artificial insemination center Singosari, Malang in Indonesia. A total of four Holstein frisian bulls of at least 3-4 years of age and average weight of 800-850 kg were used this study. The bulls were housed under controlled condition at the AI Center. The semen was collected during period from 2008 to 2013 and stored in the liquid nitrogen. The semen from several bulls was stored together in large containers containing liquid nitrogen. These containers were routinely filled with liquid nitrogen to protect that the AI straws were constantly covered by liquid nitrogen. Semen was routinely collected using artificial vagina once or twice weekly. Immediately, after the semen ejaculate was evaluated for volume, color, pH, viability, motility, abnormality and sperm concentration. The volume of each ejaculate was measured in a graduated tube. The sperm concentration was calculated with a hemocytometer.

    2.2. Semen cryopreservation

    Freshly collected semen was mixed with Tris- egg yolk extender according to Hong et al.[8]. Prior to cryopreservation, semen was diluted to obtain a final concentration of 25 x 106sperm /straw. Extended semen were loaded in 0.25 mL straws (Biovet, France) and maintained at 4 ℃ for 2 hours before freezing. Then they were then frozen at 4 cm above liquid nitrogen to achieve approximately -120 ℃ for 10 min before being immersed into liquid nitrogen, and stored for one until six year before thawing.

    2.3. Evaluation of frozen-thawed semen

    Straws were thawed in a water bath at 37 ℃ for 50 to 60 second. Thawing was done by placing the straws in a water bath at the proper temperature. Immediately after thawing, the content of each straw was emptied in a 5 mL Falcon tube at 37 ℃. The sperm suspension was kept at 37 ℃ during post-thaw incubation. A total of 140 semen straw from Friesian-Holstein bull produced during period from 2008 to 2013 and stored in the liquid nitrogen at the AI center was used to the study. Sample straw was divided into the six groups, each group consist 20 semen straws. The group one, all straw semen was produced on 2013 with storage in liquid nitrogen as long as one year, the group 2, 3, 4, 5, and 6 were produced on the 2012, 2011, 2010, 2009 and 2008 with storage in liquid nitrogen as long as 2, 3, 4, 5, and 6 year, respectively.

    2.4. Assessment motility

    The motile sperm estimate by mixing the semen gently and placing a 10 μL drop of diluted semen on a warm slide and covered with a glass cover slip (18 × 18 mm) from five selected representative fields. The mean of the five estimations was recorded as final motility score. Sperm viability was assessed using nigrosin-eosin stain[9]. Placing 10 μL drop of diluted semen on a slide and adds with 40 μL drop of nigrosin-eosin, and smears on a slide and drying quickly in heating stage (37 ℃). Microscopes were selected randomly from ten fields, with total of 200 cells. Individual sperm were recorded as being viable (unstained) or dead (stained).

    2.5. Sperm viability

    Eosin-nigrosin staining was used to evaluate sperm viability adopted from Felipe-Pérez et al [10]. After thawing, one drop of the semen was placed on a tempered glass slide, which was mixed with one drop of eosin-nigrosin solution (0.2 g of eosin and 2 g of nigrosin were dissolved in a buffered saline solution (153 mM NaCl and 9.65 mM NaH2PO4, pH=7.4), mixed for 2 hours at room temperature and filtered to obtain the staining media). The mixture was smeared on the glass slide and allowed to air dry. One hundred spermatozoa were evaluated in at least five different fields in each smear under a light microscope. Eosin penetrates in non-viable cells, which appear red, nigrosin offers a dark background facilitating the detection of viable, non-stained cells.

    2.6. Abnormalities of spermatozoa

    The abnormalities of sperm were evaluated based on classified morphological abnormalities into the categories such as loose spermatozoa head, abnormal spermatozoa head and tail formation, presence of proximal cytoplasmic droplet, or distal cytoplasmic droplet adopted from Hafez and Hafez[11].

    2.7. Statistical analysis

    Percentage of mean values (± SEM) for various parameters of semen quality during six years experimental were calculated. The statistical significances of the effects of Viability, motility and concentration after storage in liquid nitrogen were determined by ANOVA (S-PLUS Statistical Program, Insightful Corporation Seattle, WA, USA).

    3. Results

    The effect of long term storage of semen in liquid nitrogen on the motility, concentration, viability and abnormality of frozen-thawed Frisian Holstein bull Spermatozoa have been presented in Table 1 and 2. Based on the evaluation of fresh ejaculation are shown in Table 1, overall parameters of semen characteristics were considered as standard. The viability of thawed sperm (Figure 2) was not significantly different decreased (P>0.05) between storage on the 1 year and storage on the 2 years. Whereas, the viability was significantly different (P<0.05) with storage on the 3, 4, 5 and 6 years. The motility of thawed sperm was not significantly different decreased (P>0.05) on the storage 1, 2 and 3 years. Whereas, the motility was significantly different (P<0.05) with storage on the 4, 5 and 6 years. The abnormality of thawed sperm was not significantly different increased (P>0.05) on the storage 1, 2 and 3 years. Whereas, the abnormality was significantly increased (P<0.05) with storage on the 4, 5 and 6 years. On the other hand the concentration of thawed sperm was not significantly different decreased (P>0.05) during storage in liquid nitrogen as long as six years.

    Table 1 Evarage of volume, viability, motility, abnormality and concentration semen evaluation just after collected using artificial vagina on the Frisian Holstein bull.

    Table 2 Viability, motility, abnormality and concentration of sperm after thawing on storage periode in the liqiud nitrogen on the Frisian Holstein bull.

    4. Discussion

    The result of this study indicated that evarage volume of fresh ejaculate was ranged between 5.8 and 6.2 mL. This result strengthened findings reported by Shaha et al. [26] found the ejaculate volume 4.1 to 7.6 mL for Holstein Friesian cross Zebu cattle. Liquid nitrogen is used widely for the cryopreservation and long term storage of human and animal semen. This study reaffirmed previous study conducted by Hayashi and Ishobe[2] in which a similar concentration sperm of fresh ejaculate (455×106/mL) was reported in Frisian Holstein bull spermatozoa.

    Liquid nitrogen is used widely for the cryopreservation and long term storage of human and animal semen. The present of this study was to investigate the effects of sperm viability after the long storage in the liquid nitrogen. The viability and motility of thawed sperm in liquid nitrogen were gradually decreased. Decreasing viability and motility after period of storage in liquid nitrogen in the present study were caused effect of genetic and cryopreservation. Chatterjee and Gagnon[12] reported that possible damage to sperm after a number of long storage at liquid nitrogen that have been suggested genetic damage, e.g. abnormalities of chromatin structure and DNA integrity. Meanwhile, reactive oxygen species (ROS) produced from cryopreservation can also induce of damage sperm thawed[13]. Cryopreservation also is a major cause of damage to the sperm thawed[6, 7, 14]. On the other hand Mazur[15]; Mazur and Kashimoto[16] revealed that in general, storage of sperm at the liquid nitrogen is better because it prevents thermal reactions and only slow background ionizing radiations could probably have a damaging effect on sperm function over a long period of time. Fraser et al. [17] demonstrated that the prolonged storage has effects to sperm motility, mitochondrial function and plasma membrane integrity. Perdesen and Lebech, [18]; Wolley and Richardson[19] found as long as freezing and thawing, mitochondria of spermatozoa undergo damages. Mitochondria are the source of sperm energy, and damage to their structure during the cryopreservation process is associated with reduced post-thaw including sperm viability and motility[20-22].

    Another objective of this study was to determine the abnormality thawed sperm. Based on the data in Table 2 in this study, the abnormality was increase gradually during long term storage of semen in liquid nitrogen. These are probably due to deleterious effect of cryopreservation process, including cooling, freezing and thawing. Cryopreservation, freezing and thawing processes mayinduce spermatozoa damages especially to the plasma membrane and organelles[23, 24].

    Accuracy of sperm concentration in the artificial insemination (AI) has impact to production efficiency of breeding stations, product quality, and fertility[25]. The results obtained in this study show that sperm concentration after thawed was not significantly different during six years storage in the liquid nitrogen.

    Based on the results in these experiments, it may be concluded that concentration sperm during one year storage in liquid nitrogen resulted in similar concentration storage as long as six years. However, the viability and motility sperm thawed storage in liquid nitrogen during six years was lower than storage on the 1 and 2 years.

    Conflict of interest statement

    We declare that we have no conflict of interest.

    Acknowledgments

    This study was supported financially by fund of routine from Islamic University of Kalimantan. Banjarmasin, Indonesia with contract number: 041a/uniska-PP/X/2013. Authors are also grateful to Mr. Sarip Djaya and Mrs. Siti Darmawati.

    [1] Medeiros CMO, Forell F, Oliveira ATD, Rodrigues JL. Current status of sperm cryopreservation: why isn’t it better? Theriogenology 2002; 57:327-344.

    [2] Hayasi I, Isobe N. Characteristics of cryopreserved spermatozoa from a holstein-friesian bull thawed at different temperature. J Interl Develop & Cooperation 2005; 12 (1):107-110.

    [3] Correa JR. Relationships among frozen-thawed sperm characteristics assessed via the routine semen analysis, sperm functional tests and fertility of bulls in an artificial insemination program. Theriogenology 1997; 48; 721-731.

    [4] Linford E. The relationship between semen evaluation methods and fertility in the bull. J Reprod & Fertil 1976; 47; 283-291.

    [5] Hammerstedt RH, Graham JK, Nolan JP. Cryopreservation of mammalian sperm: what we ask them to survive. J Androl 1990; 11; 73-88.

    [6] Hong JHU, Wang QLI, Chen YL, Jlang ZL, Jia YH, Wang LQ, et al. Effects of addition of vitamin B12 to the extender on post- thaw motility, acrosome morphology, and plasma membrane integrity in bull semen. Turk J Veterinary & Anim Sci 2009; 35; 379-384.

    [7] Haugana TYT, Gr¨ohn E, Kommisrud E, Ropstad O. Reksen. Effects of sperm concentration at semen collection and storage period of frozen semen on dairy cow conception. Anim Reprod Sci 2007; 97:1-11.

    [8] Watson PF. Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their postthawing function. Reprod Fertil Dev 1995; 7: 871-891.

    [9] Barth AD, Oko RJ. Abnormal morphology of bovine spermatozoa. Iowa: Iowa State University Press; 1989.

    [10] Felipe-Pérez YE, Juárez-Mosqueda ML, Hernández- González EO, Valencia JJ. Viability of fersh and frozen bull sperm compared by two staining techniques. Acta Veterinaria Brasilica 2008; 2(4); 123-130.

    [11] Hafez B, Hafez ESE. Reproductive behavior. In: Reproduction in farm animals. 7ed. New York: Lippincott Williams and Wilkens;2000, p. 293-306.

    [12] Chatterjee S, Gagnon C. Production of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol Reprod Dev 2001; 59; 451-458.

    [13] Mammoto A, Masumoto N, Tahara M, Ikebuchi Y, Ohmichi M, Tasaka K, et al. Reactive oxygen species block sperm-egg fusion via oxidation of sperm sulfhydryl proteins in mice. Biol of Reprod 1996; 55: 1063-1068.

    [14] Parks JE, Graham JK. Effects of cryopreservation procedures on sperm membranes. Theriogenology 1992 ; 38:209-222.

    [15] Mazur P. Freezing of living cells: mechanisms and implications. Am J Physiol 1984; 247:C125-C142.

    [16] Mazur P, Koshimoto C. Is intracellular ice formation the cause ofdeath of mouse sperm frozen at high cooling rates. Biol Reprod 2002; 66: 1485-1490.

    [17] Fraser LJ, Strze˙zek W. Kordan. Post-thaw sperm characteristics following long-term storageof boar semen in liquid nitrogen. Anim Reprod Sci 2014 ; 147: 119 -127.

    [18] Perdesen HS, Lebech PE. Ultrastructural changes in the human spermatozoa after freezing for artificialinsemination. Fertil Steril 1971; 22: 125-133.

    [19] Wolley DM, Richardson DW. Ultrastructural injury to human spermatozoa after freezing and thawing. J Reprod Fert 1978; 53:389-394.

    [20] Gadea J. Sperm factors related to in vitro and in vivo porcine fertility. Theriogenology 2005; 63: 431- 444.

    [21] Fraser L, Strze˙zek J. Effect of different procedures of ejaculate col-lection, extenders and packages on DNA integrity of boar spermatozoafollowing freezing–thawing. Anim Reprod Sci 2007; 99: 317-329.

    [22] Ortega Ferrusola C, Sotillo-Galán Y, Varela-Fernández E, Gallardo-Bola?nos JM, González Fernández L, Tapia JA, et al. Detection of apoptosis like changes during the cryopreservation pro-cess in equine sperm. J Androl 2008 ; 29: 213-221.

    [23] Pena FJ, Johannisson A, Wallgren M, Rodriguez-Martínez H. Assessment of fresh and frozen-thawed boar semen using an annexin-v assay: a new method of evaluating sperm membrane integrity. Theriogenology 2003; 60: 677-689.

    [24] Munoz OVL, Amirat-Briand T, Diaz L, Va′squez E, Schmidt S, Desherces M, et al. Effect of semen dilution to low-sperm number per dose on motility and functionality of cryopreserved bovine spermatozoa using low-density lipoproteins (LDL) extender: Comparison to Triladyl and Bioxcell. Theriogenology 2009; 71: 895-900.

    [25] Anzar AT Kroetsch, Buhr MM. Comparison of different methods for assessment of sperm concentration and membrane integrity with bull semen. J Androl 2009; 30: 661-668.

    [26] Shaha SP, Alam MGS, Khatun M, Ahmed JU. Breeding soundness of study bulls. Bang. Veterinarian 2008; 25:51-61.

    *Corresponding author: Abdul MALIK, Department of Animal Science, Faculty of Agriculture. Islamic University of Kalimantan. Banjarmasin. Indonesia.

    E-mail: sidol_99@yahoo.com

    Foundation project: This study was supported financially by fund of routine from Islamic University of Kalimantan. Banjarmasin, Indonesia with contract number: 041a/ uniska-PP/X/2013.

    欧美精品人与动牲交sv欧美| 999精品在线视频| 啦啦啦视频在线资源免费观看| 亚洲国产中文字幕在线视频| av福利片在线| 国产黄色免费在线视频| 免费女性裸体啪啪无遮挡网站| 亚洲中文av在线| 国产精品久久久人人做人人爽| 女人高潮潮喷娇喘18禁视频| 女人被躁到高潮嗷嗷叫费观| 热99国产精品久久久久久7| 看黄色毛片网站| 多毛熟女@视频| 热99re8久久精品国产| 亚洲一卡2卡3卡4卡5卡精品中文| 精品福利永久在线观看| 90打野战视频偷拍视频| 日韩免费av在线播放| 国产激情欧美一区二区| 怎么达到女性高潮| xxxhd国产人妻xxx| svipshipincom国产片| 99香蕉大伊视频| 亚洲av第一区精品v没综合| 亚洲精品一二三| 伊人久久大香线蕉亚洲五| 欧美成人午夜精品| 国产精品乱码一区二三区的特点 | 两个人看的免费小视频| 亚洲第一欧美日韩一区二区三区| 亚洲人成电影免费在线| 大型av网站在线播放| 国产av精品麻豆| 国产男女超爽视频在线观看| 俄罗斯特黄特色一大片| 19禁男女啪啪无遮挡网站| 久久狼人影院| 咕卡用的链子| 淫妇啪啪啪对白视频| 亚洲在线自拍视频| 欧美一级毛片孕妇| 久久香蕉激情| tocl精华| 国产免费现黄频在线看| 亚洲中文av在线| 美女高潮到喷水免费观看| 两个人免费观看高清视频| 日韩 欧美 亚洲 中文字幕| 久久久精品区二区三区| 欧美不卡视频在线免费观看 | 中文字幕av电影在线播放| 亚洲欧洲精品一区二区精品久久久| 欧美成人免费av一区二区三区 | 欧美精品av麻豆av| 十分钟在线观看高清视频www| 视频区图区小说| 精品熟女少妇八av免费久了| 后天国语完整版免费观看| 久9热在线精品视频| 69精品国产乱码久久久| 亚洲九九香蕉| 久久精品人人爽人人爽视色| 国产精品香港三级国产av潘金莲| 色在线成人网| 国产亚洲av高清不卡| 两性午夜刺激爽爽歪歪视频在线观看 | 亚洲专区国产一区二区| 99久久99久久久精品蜜桃| 我的亚洲天堂| 免费av中文字幕在线| 欧美中文综合在线视频| 夜夜夜夜夜久久久久| 中文字幕色久视频| 国产精品秋霞免费鲁丝片| 国产成人欧美在线观看 | 久久国产精品影院| 成年人午夜在线观看视频| 久久精品亚洲熟妇少妇任你| 欧美性长视频在线观看| 国产亚洲精品一区二区www | 亚洲视频免费观看视频| 1024香蕉在线观看| 日韩欧美一区二区三区在线观看 | 精品国产美女av久久久久小说| 欧美av亚洲av综合av国产av| 在线视频色国产色| 国产欧美日韩一区二区三区在线| 午夜两性在线视频| 日韩中文字幕欧美一区二区| 两性午夜刺激爽爽歪歪视频在线观看 | 午夜日韩欧美国产| 精品国内亚洲2022精品成人 | 亚洲美女黄片视频| av视频免费观看在线观看| 中文字幕人妻丝袜一区二区| 久久久久久久午夜电影 | www.999成人在线观看| 国产不卡一卡二| 久久久精品免费免费高清| 黄色视频不卡| 性色av乱码一区二区三区2| 亚洲一码二码三码区别大吗| 午夜福利在线观看吧| 这个男人来自地球电影免费观看| 久久香蕉激情| 久9热在线精品视频| 午夜精品久久久久久毛片777| 欧美精品人与动牲交sv欧美| 亚洲精品中文字幕一二三四区| 精品国产乱码久久久久久男人| 亚洲精品国产精品久久久不卡| 午夜福利乱码中文字幕| 18禁观看日本| 麻豆成人av在线观看| 成年版毛片免费区| 色综合婷婷激情| 久久久久久久久免费视频了| 精品久久蜜臀av无| 国产欧美日韩综合在线一区二区| 欧美精品人与动牲交sv欧美| 国产精品av久久久久免费| 91精品三级在线观看| 巨乳人妻的诱惑在线观看| 久久久久久久国产电影| 亚洲精品成人av观看孕妇| 国产精品国产高清国产av | av线在线观看网站| 纯流量卡能插随身wifi吗| 久久久精品区二区三区| 在线观看免费视频日本深夜| 岛国在线观看网站| 国产精品乱码一区二三区的特点 | 91麻豆精品激情在线观看国产 | 亚洲五月色婷婷综合| 动漫黄色视频在线观看| 日韩免费av在线播放| 欧美在线黄色| 国产一区二区三区综合在线观看| 少妇裸体淫交视频免费看高清 | 国产精品乱码一区二三区的特点 | 丝袜美腿诱惑在线| 满18在线观看网站| 男人的好看免费观看在线视频 | 午夜两性在线视频| 国产在线一区二区三区精| 三上悠亚av全集在线观看| 91成人精品电影| 日韩制服丝袜自拍偷拍| 嫩草影视91久久| 日韩制服丝袜自拍偷拍| 午夜福利一区二区在线看| videosex国产| 人人妻人人澡人人爽人人夜夜| 久久香蕉激情| 99精国产麻豆久久婷婷| 美女高潮到喷水免费观看| 久久中文看片网| 成年动漫av网址| 日韩欧美三级三区| 一二三四社区在线视频社区8| 午夜免费观看网址| 天堂动漫精品| 电影成人av| 亚洲国产精品一区二区三区在线| 精品福利永久在线观看| 一进一出抽搐动态| 可以免费在线观看a视频的电影网站| 久久国产精品人妻蜜桃| 欧美最黄视频在线播放免费 | 欧美日韩福利视频一区二区| 亚洲五月婷婷丁香| 中国美女看黄片| 中亚洲国语对白在线视频| 丝袜人妻中文字幕| 亚洲人成电影观看| 99riav亚洲国产免费| 久久久久久久国产电影| 韩国精品一区二区三区| 国产精品香港三级国产av潘金莲| 精品乱码久久久久久99久播| 中文字幕人妻丝袜一区二区| а√天堂www在线а√下载 | 99久久99久久久精品蜜桃| 女人爽到高潮嗷嗷叫在线视频| 女性被躁到高潮视频| 建设人人有责人人尽责人人享有的| 人人妻,人人澡人人爽秒播| 99久久人妻综合| 国产一区有黄有色的免费视频| 亚洲av美国av| 日本a在线网址| 亚洲aⅴ乱码一区二区在线播放 | 久久精品国产99精品国产亚洲性色 | 99在线人妻在线中文字幕 | 性少妇av在线| 精品午夜福利视频在线观看一区| 精品国内亚洲2022精品成人 | videos熟女内射| 一区福利在线观看| 视频区欧美日本亚洲| 日本vs欧美在线观看视频| 精品高清国产在线一区| 国产精品98久久久久久宅男小说| 乱人伦中国视频| 男女之事视频高清在线观看| 久久草成人影院| 亚洲一区二区三区不卡视频| 99久久人妻综合| 国产一区二区三区在线臀色熟女 | 国产男靠女视频免费网站| 国产蜜桃级精品一区二区三区 | netflix在线观看网站| 少妇粗大呻吟视频| 国产精品国产av在线观看| 最近最新中文字幕大全免费视频| 51午夜福利影视在线观看| 久久久久国内视频| 精品欧美一区二区三区在线| 亚洲 欧美一区二区三区| 国产在线观看jvid| 欧美黑人精品巨大| 国产有黄有色有爽视频| 欧美色视频一区免费| 伦理电影免费视频| 欧美最黄视频在线播放免费 | 嫁个100分男人电影在线观看| 一区在线观看完整版| 18禁裸乳无遮挡动漫免费视频| 色尼玛亚洲综合影院| 在线观看免费高清a一片| 欧美 亚洲 国产 日韩一| 国产视频一区二区在线看| 欧美日韩亚洲高清精品| 成人国产一区最新在线观看| 午夜福利欧美成人| 国产高清激情床上av| 99在线人妻在线中文字幕 | 精品一区二区三区av网在线观看| 亚洲欧美一区二区三区久久| 欧美精品啪啪一区二区三区| 动漫黄色视频在线观看| 波多野结衣av一区二区av| 国产精品免费一区二区三区在线 | 中文欧美无线码| 日韩一卡2卡3卡4卡2021年| 亚洲第一欧美日韩一区二区三区| 怎么达到女性高潮| 又黄又粗又硬又大视频| 亚洲精品在线美女| av中文乱码字幕在线| 久久天堂一区二区三区四区| 久久久国产精品麻豆| 久久亚洲真实| 午夜老司机福利片| 亚洲综合色网址| 精品一区二区三区av网在线观看| 国产免费男女视频| 久久婷婷成人综合色麻豆| 国产xxxxx性猛交| 亚洲第一青青草原| 性色av乱码一区二区三区2| 在线观看免费日韩欧美大片| 最新在线观看一区二区三区| 看免费av毛片| 精品福利观看| 极品少妇高潮喷水抽搐| 一本一本久久a久久精品综合妖精| 成在线人永久免费视频| 最近最新免费中文字幕在线| 天堂√8在线中文| 99国产极品粉嫩在线观看| 精品国产美女av久久久久小说| 90打野战视频偷拍视频| 亚洲国产精品一区二区三区在线| 欧美日韩一级在线毛片| 人妻久久中文字幕网| 久久热在线av| 色老头精品视频在线观看| 女人被躁到高潮嗷嗷叫费观| 亚洲国产看品久久| 日韩熟女老妇一区二区性免费视频| 电影成人av| 久久ye,这里只有精品| 日本wwww免费看| 国产成人一区二区三区免费视频网站| 国产精品欧美亚洲77777| 午夜福利影视在线免费观看| 国产高清视频在线播放一区| 亚洲片人在线观看| 国产精品.久久久| 又大又爽又粗| 首页视频小说图片口味搜索| 国产激情欧美一区二区| 亚洲五月色婷婷综合| 国产一区有黄有色的免费视频| 国产精品九九99| 麻豆国产av国片精品| 搡老乐熟女国产| 又紧又爽又黄一区二区| 在线观看一区二区三区激情| 国产成人精品在线电影| 18禁黄网站禁片午夜丰满| 国产精品电影一区二区三区 | 欧美黄色片欧美黄色片| 国产精品久久视频播放| www.熟女人妻精品国产| 国产精品国产高清国产av | bbb黄色大片| 亚洲精品久久午夜乱码| 成人特级黄色片久久久久久久| 久久精品aⅴ一区二区三区四区| 欧美日韩亚洲综合一区二区三区_| 久久香蕉激情| 搡老乐熟女国产| 777久久人妻少妇嫩草av网站| 色在线成人网| videosex国产| 香蕉国产在线看| 精品久久久久久久久久免费视频 | 免费av中文字幕在线| 欧美日韩瑟瑟在线播放| 久久久国产欧美日韩av| 精品视频人人做人人爽| 亚洲中文av在线| 麻豆国产av国片精品| 久久热在线av| 动漫黄色视频在线观看| 欧美午夜高清在线| 久久香蕉精品热| 久热这里只有精品99| 一级片免费观看大全| 91麻豆av在线| 欧美性长视频在线观看| 国产xxxxx性猛交| 亚洲aⅴ乱码一区二区在线播放 | 少妇被粗大的猛进出69影院| 午夜亚洲福利在线播放| 久久久国产成人精品二区 | 91九色精品人成在线观看| 十八禁高潮呻吟视频| 性色av乱码一区二区三区2| 中文字幕色久视频| 国产亚洲精品一区二区www | 日本撒尿小便嘘嘘汇集6| 色播在线永久视频| 亚洲精品在线观看二区| 亚洲人成77777在线视频| 69精品国产乱码久久久| 在线观看免费高清a一片| 嫩草影视91久久| 黄频高清免费视频| 精品欧美一区二区三区在线| 在线观看66精品国产| 18禁国产床啪视频网站| 久久天躁狠狠躁夜夜2o2o| 成人18禁在线播放| 久久国产精品影院| 国产不卡av网站在线观看| 一区在线观看完整版| 久久人妻av系列| 欧美日韩亚洲高清精品| 91成年电影在线观看| 日韩一卡2卡3卡4卡2021年| 高清在线国产一区| 国产精品99久久99久久久不卡| 国产男靠女视频免费网站| 精品人妻熟女毛片av久久网站| 中文字幕人妻熟女乱码| 啦啦啦在线免费观看视频4| 涩涩av久久男人的天堂| 黄频高清免费视频| 国产aⅴ精品一区二区三区波| 婷婷成人精品国产| 啦啦啦 在线观看视频| 国产成人av教育| 咕卡用的链子| 国产又色又爽无遮挡免费看| 中文欧美无线码| 热re99久久国产66热| 国产精品香港三级国产av潘金莲| 久久天躁狠狠躁夜夜2o2o| 国产成人av激情在线播放| 纯流量卡能插随身wifi吗| 精品久久久久久久毛片微露脸| 777米奇影视久久| av免费在线观看网站| 18禁国产床啪视频网站| 免费在线观看黄色视频的| 欧美日韩瑟瑟在线播放| 精品乱码久久久久久99久播| 久99久视频精品免费| 一本综合久久免费| 国产淫语在线视频| 大香蕉久久成人网| 色播在线永久视频| 亚洲国产看品久久| 午夜精品在线福利| 色播在线永久视频| 国产淫语在线视频| 欧美亚洲 丝袜 人妻 在线| 99精品在免费线老司机午夜| 久久精品国产a三级三级三级| 亚洲欧美一区二区三区黑人| www.熟女人妻精品国产| 久久婷婷成人综合色麻豆| 亚洲国产精品一区二区三区在线| 多毛熟女@视频| 激情视频va一区二区三区| 国产三级黄色录像| 老司机午夜十八禁免费视频| 免费日韩欧美在线观看| 在线永久观看黄色视频| 欧美精品一区二区免费开放| 国产99久久九九免费精品| 在线观看一区二区三区激情| 亚洲五月婷婷丁香| 王馨瑶露胸无遮挡在线观看| 免费少妇av软件| 亚洲五月婷婷丁香| 精品卡一卡二卡四卡免费| 80岁老熟妇乱子伦牲交| 日本vs欧美在线观看视频| 一级毛片高清免费大全| 久久狼人影院| 99香蕉大伊视频| 亚洲午夜精品一区,二区,三区| 91精品三级在线观看| 精品久久蜜臀av无| 中文亚洲av片在线观看爽 | www.熟女人妻精品国产| 少妇粗大呻吟视频| 国产伦人伦偷精品视频| 亚洲精品国产色婷婷电影| 国产欧美日韩一区二区三区在线| 国产成人精品久久二区二区91| 久久影院123| 新久久久久国产一级毛片| 在线观看午夜福利视频| 免费一级毛片在线播放高清视频 | 三级毛片av免费| 成年版毛片免费区| 国产精品免费一区二区三区在线 | 看免费av毛片| 欧美在线黄色| 一区在线观看完整版| 在线十欧美十亚洲十日本专区| a级片在线免费高清观看视频| 中文欧美无线码| 亚洲av片天天在线观看| 一区福利在线观看| 精品熟女少妇八av免费久了| 两性午夜刺激爽爽歪歪视频在线观看 | 免费看a级黄色片| 欧美日韩亚洲高清精品| 一二三四社区在线视频社区8| 天堂中文最新版在线下载| 91大片在线观看| 欧美亚洲日本最大视频资源| 国产av精品麻豆| www.熟女人妻精品国产| 纯流量卡能插随身wifi吗| av网站免费在线观看视频| 黄色怎么调成土黄色| 亚洲欧美色中文字幕在线| a级毛片在线看网站| 亚洲午夜理论影院| 国产淫语在线视频| 亚洲欧美一区二区三区黑人| 成人三级做爰电影| 精品少妇一区二区三区视频日本电影| 天天影视国产精品| 国产色视频综合| 国产在视频线精品| 日韩欧美三级三区| 色综合欧美亚洲国产小说| 亚洲精品中文字幕在线视频| 亚洲一码二码三码区别大吗| 一级毛片高清免费大全| 亚洲国产毛片av蜜桃av| 嫁个100分男人电影在线观看| 中文字幕精品免费在线观看视频| 啦啦啦 在线观看视频| 久久ye,这里只有精品| 久久久国产欧美日韩av| 后天国语完整版免费观看| 手机成人av网站| 黑丝袜美女国产一区| 精品第一国产精品| 欧美在线黄色| 久久这里只有精品19| 亚洲三区欧美一区| 满18在线观看网站| 看黄色毛片网站| 天堂√8在线中文| 丰满人妻熟妇乱又伦精品不卡| 村上凉子中文字幕在线| 国产国语露脸激情在线看| 麻豆乱淫一区二区| 国产精品偷伦视频观看了| 少妇 在线观看| cao死你这个sao货| 人人妻人人添人人爽欧美一区卜| 色在线成人网| 久久香蕉激情| 久久国产精品大桥未久av| 中文字幕最新亚洲高清| 日本欧美视频一区| 国产成人免费观看mmmm| 成人国产一区最新在线观看| tocl精华| 欧美成人午夜精品| 欧美精品人与动牲交sv欧美| 国产精品久久久久成人av| 久久人妻福利社区极品人妻图片| 男男h啪啪无遮挡| 亚洲成人国产一区在线观看| 免费久久久久久久精品成人欧美视频| 老司机靠b影院| 在线看a的网站| 不卡av一区二区三区| 在线天堂中文资源库| 最近最新中文字幕大全电影3 | 妹子高潮喷水视频| 青草久久国产| 精品国产超薄肉色丝袜足j| 久久性视频一级片| 亚洲欧美激情在线| 两性夫妻黄色片| 热99国产精品久久久久久7| 18禁美女被吸乳视频| av不卡在线播放| 精品国产乱码久久久久久男人| 亚洲午夜理论影院| 日韩视频一区二区在线观看| 黑人欧美特级aaaaaa片| av欧美777| 免费av中文字幕在线| 国产xxxxx性猛交| 欧美日韩亚洲国产一区二区在线观看 | 天天躁日日躁夜夜躁夜夜| 黄色视频,在线免费观看| 美国免费a级毛片| 国内毛片毛片毛片毛片毛片| 美国免费a级毛片| 91av网站免费观看| 美国免费a级毛片| 国产有黄有色有爽视频| 中文字幕精品免费在线观看视频| 51午夜福利影视在线观看| 日韩免费高清中文字幕av| 大型黄色视频在线免费观看| 在线观看午夜福利视频| 国产精品1区2区在线观看. | 中文欧美无线码| 亚洲国产欧美日韩在线播放| 亚洲五月婷婷丁香| 成人特级黄色片久久久久久久| 国产精品久久久久久人妻精品电影| 亚洲午夜精品一区,二区,三区| 免费人成视频x8x8入口观看| 成年版毛片免费区| 国产精品久久久久久人妻精品电影| 手机成人av网站| 精品福利永久在线观看| 手机成人av网站| 亚洲av欧美aⅴ国产| 曰老女人黄片| 亚洲av欧美aⅴ国产| a级毛片黄视频| 性色av乱码一区二区三区2| 久久精品国产综合久久久| 精品久久久久久久毛片微露脸| 高清欧美精品videossex| 亚洲精品中文字幕一二三四区| 亚洲国产欧美网| 午夜福利免费观看在线| 99精品在免费线老司机午夜| 国产三级黄色录像| 亚洲av成人一区二区三| 久久人妻熟女aⅴ| 国产精品国产高清国产av | 曰老女人黄片| 欧美日韩成人在线一区二区| 婷婷成人精品国产| 精品久久久久久电影网| 午夜影院日韩av| 夜夜夜夜夜久久久久| 久久狼人影院| 两个人看的免费小视频| 19禁男女啪啪无遮挡网站| 久久精品国产综合久久久| 在线看a的网站| 午夜激情av网站| 十八禁人妻一区二区| 丝袜在线中文字幕| 精品久久久久久电影网| tube8黄色片| 人妻久久中文字幕网| 一区二区日韩欧美中文字幕| 亚洲精品国产色婷婷电影| 亚洲成人国产一区在线观看| 怎么达到女性高潮| 人人妻人人澡人人爽人人夜夜| 亚洲专区字幕在线| 两个人看的免费小视频| 黄色视频不卡| 咕卡用的链子| 夜夜爽天天搞| 不卡一级毛片| 精品无人区乱码1区二区| 久久国产乱子伦精品免费另类| 国产精品亚洲av一区麻豆| 在线视频色国产色| 亚洲精品国产一区二区精华液|