• <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 ?

    Experimental infectivity of Theilerialuwenshuni and Theileria uilenbergi in Chinese Kunming mice

    2018-02-05 07:11:01LIYouquanGUOPengfeiLIUJunlongLIUZhijieHANYuanLIXuanLIUAihongGUANGuiquanLIUGuangyuanLUOJianxunYINHong
    Journal of Integrative Agriculture 2018年2期

    LI You-quan, GUO Peng-fei LIU Jun-long LIU Zhi-jie HAN Yuan LI Xuan LIU Ai-hong GUAN Gui-quan, LIU Guang-yuan, LUO Jian-xun, YIN Hong

    1 State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, P.R.China

    2 Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, P.R.China

    1. Introduction

    Theileriaparasites are transmitted by tick species of the familyIxodidaeand cause significant losses to the cattle and sheep industry in endemic areas (Luo and Lu 1997; Aktaset al. 2004; Simuunzaet al. 2011). SomeTheileriaspecies that infect sheep and goats are pathogenic, includingTheileria lestoquardi,Theileria uilenbergi, andTheileria luwenshuni(Luo and Yin 1997; Schnitteret al. 2000; Yinet al. 2007).T.luwenshuniandT.uilenbergiare widely distributed in northern China (Luo and Yin 1997; Yin 2000; Liet al. 2014c) and are transmitted by the ticksHaemaphysalis qinghaiensisandHaemaphysalis longicornisin China(Yin 2000; Yinet al. 2002; Liet al. 2007, 2009, 2014c).T.luwenshuniinfects sheep from Great Britain (Phippset al.2016).T.luwenshuniandT.ulenbergiwere believed to infect only sheep and goats (Luo and Yin 1997; Yin 2000;Yinet al. 2007).

    However, a recent study has reported thatT.luwenshuniinfects several animal species, including cervids (Hanet al.2009; Liet al. 2014a, 2015; Leeet al. 2016), Mongolian gazelle (Liet al. 2014b),hedgehog (Chenet al. 2014), cattle, and yak (Qinet al. 2016).T.uilenbergican also infect several cervid species, including sika deer, red deer, and roe deer (Liet al. 2014a, 2015), as well as cattle, yak, and cattle-yak (unpublished data). In the 1990s, some studies reported that mice were used for the isolation ofBabesiaparasites from grazing calves in Japan (Tsujiet al. 1995),Theileria annulataschizonts infected severe combined immunodeficiency (SCID) mice (Fellet al. 1990), andTheileria sergentiinfected SCID mice with bovine erythrocyte transfusion (Tsujiet al. 1992). The Chinese Kunming mouse strain (Mus musculusKunming, KM), an outbreed mouse strain originating from the Swiss Albino mouse,is widely used in pharmacological and gene-related studies throughout China (Yanet al. 2017). Kunming mice are widely used as experimental animals in China because it exhibits many advantages such as high disease resistance, large and frequent litters and rapid growth rate. However, no previous studies have evaluated the infectivity ofT.luwenshuniandT.uilenbergiin mice. Therefore, the present study aimed at determining the experimental infectivity ofT.luwenshuniandT.uilenbergiin Kunming mice by microscopic examination (ME) and PCR analysis.

    2. Materials and methods

    This study was approved by the Animal Research Ethics Committee of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (No.LVRIAEC2012-003). All sheep and mice were handled in accordance with good animal practices found in the Animal Ethics Procedures and Guidelines of the People’s Republic of China.

    2.1. Parasites

    T.luwenshuniwas isolated from goats from Weiyuan County, Gansu Province,China in 2005 and identified as a singleT.luwenshunispecies based on morphological, transmission and molecular methods.T.uilenbergiwas isolated from sheep from Yongjing County, Gansu Province, China in 2012 and identified as a singleT.uilenbergispecies based on morphological, transmission, and molecular method. The twoTheileriaspp. isolates were cryopreserved in liquid nitrogen at OIE Reference Laboratory for Ovine Theileriosis of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences.

    2.2. Sheep

    Two 5-month-old sheep from Jingtai County, Gansu Province were identified to beTheileria-free andBabesia-free using the methods described by Yin (2000),and utilized in this study.

    2.3. Inoculation of mice with T. luwenshuni and T. uilenbergi

    ?

    Sheep No. 11 was inoculated withT.luwenshuniisolate, and sheep No. 22 was inoculated withT.uilenbergiisolate. When the parasitemia of sheep No. 11 reached 5% and the parasitemia of sheep No. 22 reached 4%, the blood samples of the two sheep were individually collected into anticoagulant tubes and utilized in inoculation. Five mice were used as the control group. In theT.luwenshuniexperimental group, 20 Chinese Kunming mice divided into four groups (Table 1):(1) five splenectomized mice were inoculated intraperitoneally with 0.4 mL ofTheileria-infected blood, (2) five unsplenectomized mice were inoculated intraperitoneally with 0.4 mL ofTheileria-infected blood, (3) five splenectomized mice were inoculated intravenously with 0.2 mL ofTheileria-infected blood, and (4)five unsplenectomized mice were inoculated intravenously with 0.2 mL ofTheileria-infected blood. In theT.uilenbergiexperimental group, the arrangement is similar to theT.luwenshuniexperimental group. Total of 100 μL whole blood was collected into 4 mL blood collection tube containing K3EDTA from the tails of 45 Chinese Kunming mice each 4 days, respectively. Genomic DNA of 45 mice whole blood samples was extracted individually using DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany)according to the manufacturer’s instructions. The concentration of DNA was determined using a NanoDrop ND-2000 Spectrophotometer (Nanodrop Technologies,Wilmington, DE, USA). In addition, the blood smears of 45 mice were prepared using mouse caudal vein blood each 2 days, respectively. Ten days after inoculation, the blood smears were analyzed by microscopic examination every 2 days and genomic DNA was screened by PCR analysis every 4 days. After that, all mice were monitored every 2 days for 3 months by measuring two physiological indexes:red blood cells (RBC) and hemoglobin (Hb).

    2.4. Microscopic analysis of blood smears

    The blood smears were air-dried, fixed in methanol, stained with a 10% solution of Giemsa in deionized water (pH 7.2),and analyzed microscopically.

    2.5. Molecular detection of blood parasites

    PCR was used to detectTheileriaspp. in Chinese Kunming mice using genus-specific primers (Allosopet al. 1993) and species-specific primers (Yin 2000;Yinet al. 2008). The specific primers ofT.luwenshuniare Tl-f: GGTAGGGTATTGGCCTACTGA and Tl-r:TCATCCGGATAATACAAGT, with a 340-bp product (Yin 2000; Yinet al. 2008); the specific primers ofT.uilenbergiare Tu-f: GGTAGGGTATTGGCCTACCGG and Tu-r:ACACTCGGAAAATGCAAGCA, with a 340-bp products(Yin 2000; Yinet al. 2008). The specific primers ofTheileriaspp. are 989-f: AGTTTCTGACCTATCAG and 990-r: TTGCCTTAAACTTCCTTG, with a 1 100-bp product(Allosopet al. 1993). The PCRs were performed in an automatic DNA thermocycler (Bio-Rad, Hercules, CA, USA)according to the previously described methods (Allosopet al. 1993; Yin 2000; Yinet al. 2008), and the PCR products were separated by 1.5% agarose gel electrophoresis to assess the presence of specific fragments indicative of the presence ofTheileria. After that, the amplification products were sequenced by GenScript Corporation (Piscataway,NJ, USA), and representative sequences of the 18S rDNA genes ofT.luwenshuniandT.uilenbergiwere analyzed using Basic Local Alignment Search TooL (BLAST) from the National Center for Biotechnology Information (NCBI)(http://www.ncbi.nlm.nih.gov/Blast.cgi). The 18S rRNA gene sequences ofTheileriaspp. obtained from Chinese Kunming mice in this study were deposited in GenBank.

    3. Results and discussion

    On days 18 to 20,Theileriainfection was detected microscopically in 40 mice from the experimental groups,and the levels of parasitemia in all infected mice were very low, ranging between 0.01 and 0.03% (Fig. 1). NoTheileriaparasites were detected in the control group during the study period. On days 16 to 18,T.luwenshuni-specific DNA fragments were detected in the fourT.luwenshuniinoculated mice groups, andT.uilenbergi-specific DNA fragments were detected in the fourT.ulenbergi-inoculated mice groups. However, noTheileriaparasites were detected in blood samples from the control group by ME and PCR.BLAST analysis indicated that theT.luwenshuni-specific DNA sequences in the experimental groups shared 100%similarity with the sequences fromT.luwenshuniisolate Weiyuan; meanwhile,T.uilenbergi-specific DNA sequences in the experimental groups also shared 100% similarity with the sequences fromT.uilenbergiisolate Yongjing.The 18S rRNA gene sequences ofTheileriaspp. obtained from Chinese Kunming mice in this study were deposited in GenBank under the following accession numbers:KY508396-KY508399 forT.luwenshuni, and KY508400-KY508403 forT.uilenbergi. The RBC count and Hb levels of the 40 infected mice had no obvious changes in the course of infection. In this study, there is not obviously observed changes of the parasitemia rate of the infected Kunming mice between the species ofT. luwenshuniandT. uilenbergi, or injectionviacaodal wein and enterocoelia.Two reasons are for the low parastemia in mice: One is that the mice are not suitable hosts forT.luwenshuniandT.uilenbergi; another is thatT.luwenshuniandT.uilenbergiare not pathogenic to mice so that all inoculated mice have no hemolytic anemia phenomenon.

    Fig. 1 Theileria luwenshuni parasite in red blood cells of a Chinese Kunming mouse (see arrow).

    Tsujiet al. (1995) used a Bo-RBC-SCID mouse model to isolate theBabesiaparasites from grazing calves in Japan,and Bo-RBC-SCID mice infected withBabesiaparasites developed a high parasitemia and presented with severe clinical symptoms, including hemoglobinuria, jaundice,and hemolytic anemia. Fellet al. (1990) reported that mice injected subcutaneously withT. annulataschizonts developed solid tumors at the injection site, whereas those injected intraperitoneally with these parasites developed ascites. Tsujiet al. (1992) reported the first successful propagation ofT.sergentiin SCID mice supplied periodically with uninfected bovine erythrocytes. Hagiwaraet al. (1993)identified parasite infection in mice inoculated intravenously,intraperitoneally, and subcutaneously. However, the animals inoculated intravenously developed parasitemia earlier than those inoculated intraperitoneally or subcutaneously. Herein we found that the level of parasitemia in the normal Chinese Kunming mice inoculated intravenously was not significantly different from that in mice inoculated intraperitoneally, the possible reason is that the low parasitemia of the infected normal mice influences the difference.

    AlthoughT.luwenshuniandT.uilenbergiinfected mice,the infected mice had no obviously pathological changes(e.g., hemolytic anemia). In addition, the parasitemia of these two parasite species was low in mice. Therefore,the Kunming mouse is not a good model of infection with the twoTheileria. However, further studies are needed to evaluate whetherT.luwenshuniandT.uilenbergican efficiently infect SCID mice with jaundice and hemolytic anemia. Furthermore, the elucidation of the role of normal mice as potential reservoir hosts forT.luwenshuniandT.uilenbergiis critical for determining whether they help spread theileriosis to wild ruminants.

    4. Conclusion

    Our results demonstrated the occurrence of multi-homing parasitism inT.luwenshuniandT.uilenbergiin lab, which were believed to infect only sheep and goats. AlthoughT.luwenshuniandT.uilenbergican infect normal Kunming mice, the Kunming mouse is not suitable for a good model of infection withTheileriaparasites because they did not develop high parasitemia and showed no severe clinical symptoms such as jaundice and hemolytic anemia.

    Acknowledgements

    This study was financially supported by the National Key Research and Development Program of China(2017YFD0501200, 2016YFC1202000, 2016YFC1202002),the earmarked fund for China Agriculture Research System(CARS-37), the National Natural Science Foundation of China (31272556, 31402189, 31372432), the Agricultural Science and Technology Innovation Program, China(2014ZL010), the National Basic Research Program of China (2015CB150300), the Special Funds for Agroscientific Research in the Public Research, China(201303035), the Gansu International Collaboration Special Project, China (1504WKCA056), the Jiangsu Co-Innovation Center Programme for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, State Key Laboratory of Veterinary Etiological Biology Project, China.We are grateful to International Science Editing (Ireland) for editing the manuscript.

    Aktas M, Dumanli N, Angin M. 2004. Cattle infestation byHyalommaticks and prevalence ofTheileriainHyalommaspecies in the east of Turkey.Veterinary Parasitology,119, 1–8.

    Allsopp B A, Baylis H A, Allsopp M T, Cavalier-Smith T, Bishop R P, Carrington D M, Sohanpal B, Spooner P. 1993.Discrimination between six species ofTheileriausing oligonucleotide probes which detect small subunit ribosomal RNA sequences.Parasitology, 107, 157–165.

    Chen Z, Liu Q, Jiao F C, Xu B L, Zhou X N. 2014. Detection of piroplasms infection in sheep, dogs and hedgehogs in Central China.Infectious Diseases of Poverty, 3, doi:10.1186/2049-9957-3-18

    Fell A H, Preston P M, Ansell J D. 1990. Establishment ofTheileria-infected bovine cell lines inscidmice.Parasite Immunology, 12, 335–359.

    Hagiwara K, Tsuji M, Ishihara C, Tajima M, Kurosawa T, Iwai H, Takahashi K. 1993.Theileria sergentiinfection in the Bo-RBC-SCID mouse model.Parasitology Research, 79,466–470.

    Han J, Jang H, Lee S, Na K. 2009. High prevalence ofTheileriasp. in wild Chinese water deer (Hydropotes inermis argyropus) in South Korea.Veterinary Parasitology, 164,311–314.

    Lee S H, Kim K T, Kwon O D, Ock Y, Kim T, Choi D, Kwak D.2016. Novel detection ofCoxiellaspp.,Theileria luwenshuni,andT.ovisendosymbionts in deer keds(Lipoptena fortisetosa).PLoS ONE, 11, e0156727.

    Li Y, Chen Z, Liu Z, Liu J, Yang J, Li Q, Li Y, Cen S, Guan G, Ren Q, Luo J, Yin H. 2014a. Molecular identification ofTheileriaparasites of northwestern Chinese Cervidae.Parasites & Vectors, 14, 225.

    Li Y, Chen Z, Liu Z, Liu J, Yang J, Li Q, Li Y, Ren Q, Niu Q,Guan G, Luo J, Yin H. 2014b. First report ofTheileriaandAnaplasmain the Mongolian gazelle,Procapra gutturosa.Parasites & Vectors, 7, 614.

    Li Y, Liu J, Liu Z, Yang J, Li Y, Li Q, Qin G, Chen Z, Guan G, Luo J, Yin H. 2015. Report ofTheileria luwenshuniandTheileriasp. RSR from cervids in Gansu, China.Parasitology Research, 114, 2023–2029.

    Li Y, Luo J, Guan G, Ma M, Liu A, Liu J, Ren Q, Niu Q, Lu B,Gao J, Liu Z, Dang Z, Tian Z, Zhang B, He Z, Bai Q, Yin H. 2009. Experimental transmission ofTheileria uilenbergiinfective for small ruminants byHaemaphysalis longicornisandHaemaphysalis qinghaiensis.Parasitology Research,104, 1227–1231.

    Li Y, Luo J, Liu Z, Guan G, Gao J, Ma M, Dang Z, Liu A, Ren Q, Lu B, Liu J, Zhao H, Li J, Liu G, Bai Q, Yin H. 2007.Experimental transmission ofTheileriasp. (China 1)infective for small ruminants byHaemaphysalis longicornisandHaemaphysalis qinghaiensis.Parasitology Research,101, 533–538.

    Li Y, Zhang X, Liu Z, Chen Z, Yang J, He H, Guan G, Liu A, Ren Q, Niu Q, Liu J, Luo J, Yin H. 2014c. An epidemiological survey ofTheileriainfections in small ruminants in central China.Veterinary Parasitology, 200, 198–202.

    Luo J, Lu W. 1997. Cattle theileriosis in China.Tropical Animal Health and Production, 29(Suppl. 4), 4S–7S.

    Luo J, Yin H. 1997. Theileriosis of sheep and goats in China.Tropical Animal Health and Production, 29, 8s–9s.

    Phipps L P, Hernández-Triana L M, Goharriz H, Welchman D, Johnson N. 2016. Detection ofTheileria luwenshuniin sheep from Great Britain.Parasites & Vectors, 9, 203.

    Qin G, Li Y, Liu J, Liu Z, Yang J, Zhang L, Liu G, Guan G, Luo J, Yin H. 2016. Molecular detection and characterization ofTheileriainfection in cattle and yaks from Tibet Plateau Region, China.Parasitology Research, 115, 2647–2652.

    Schnitter L, Yin H, Jianxun L, Ludwig W, Shayan P, Rahbari S, Voss-Holtmann A, Ahmed J S. 2000. Ribosomal small-subunit RNA gene-sequence analysis ofTheileria lestoquardiand aTheileriaspecies highly pathogenic forTheileria buffeli(synT.orientalis) in cattle.Research in Veterinary Science, 86, 352–358.

    Simuunza M, Weir W, Courcier E, Tait A, Shiels B. 2011.Epidemiological analysis of tick-borne diseases in Zambia.Veterinary Parasitology, 175, 331–342.

    Tsuji M, Hagiwara K, Takahashi K, Ishihara C, Azuma I, Siddiqui W A. 1992.Theileria sergentiproliferates in SCID mice with bovine erythrocyte transfusion.Journal of Parasitology,78, 750–752.

    Tsuji M, Terada Y, Arai S, Okada H, Ishihara C. 1995. Use of the Bo-RBC-SCID mouse model for isolation of aBabesiaparasite from grazing calves in Japan.Experimental Parasitology, 81, 512–518.

    Yan W, Yao L, Liu W, Sun K, Zhang Z, Zhang L. 2017. A kind of rd1 mouse in C57BL/6J mice from crossing with a mutated Kunming mouse.Gene, 607, 9–15.

    Yin H. 2000. Biological and molecular characterization ofTheileriasp. infective to sheep and goats. Ph D thesis,Graduate School of Chinese Academy of Agricultural Sciences, China. pp. 6–122. (in Chinese)

    Yin H, Guan G Q, Ma M L, Luo J X, Lu B Y, Yuan G, Bai Q, Lu C, Yuan Z, Preston P. 2002.Haemaphysalis qinghaiensisticks transmit at least two differentTheileriaspecies: One is infective to yaks, one is infective to sheep.Veterinary Parasitology, 107, 29–35.

    Yin H, Liu Z, Guan G, Liu A, Ma M, Ren Q, Luo J. 2008. Detection and differentiation ofTheileria luwenshuniandT.uilenbergiinfection in small ruminants by PCR.Transboundary and Emerging Diseases, 55, 233–237.

    Yin H, Schnittger L, Luo J X, Ulrike S, Ahmed J. 2007. Ovine theileriosis in China: A new look at an old story.Parasitology Research, 101(Suppl. 2), S191–S195.

    成年人黄色毛片网站| 侵犯人妻中文字幕一二三四区| 欧美丝袜亚洲另类 | 看片在线看免费视频| 午夜亚洲福利在线播放| 日本黄色日本黄色录像| 宅男免费午夜| 亚洲成人国产一区在线观看| 亚洲视频免费观看视频| 夜夜爽天天搞| 国内久久婷婷六月综合欲色啪| netflix在线观看网站| 亚洲色图av天堂| 欧美一区二区精品小视频在线| 午夜免费观看网址| 国产成人影院久久av| e午夜精品久久久久久久| av国产精品久久久久影院| 亚洲人成网站在线播放欧美日韩| avwww免费| 国内毛片毛片毛片毛片毛片| 在线av久久热| 日韩国内少妇激情av| 亚洲 欧美 日韩 在线 免费| 精品乱码久久久久久99久播| 高清黄色对白视频在线免费看| 在线观看日韩欧美| 免费观看精品视频网站| 老汉色∧v一级毛片| 精品人妻在线不人妻| 黄频高清免费视频| 欧美成人免费av一区二区三区| 精品国产国语对白av| 麻豆久久精品国产亚洲av | 日韩大尺度精品在线看网址 | 村上凉子中文字幕在线| 中文字幕另类日韩欧美亚洲嫩草| 亚洲人成伊人成综合网2020| 国产亚洲精品一区二区www| 好男人电影高清在线观看| 免费高清视频大片| 精品熟女少妇八av免费久了| 久久 成人 亚洲| 亚洲 欧美 日韩 在线 免费| 黄色视频不卡| 欧美日韩一级在线毛片| 一级毛片高清免费大全| 午夜影院日韩av| 美女高潮喷水抽搐中文字幕| 制服诱惑二区| 亚洲精品久久午夜乱码| 亚洲精品在线美女| 久久久水蜜桃国产精品网| 在线观看免费日韩欧美大片| av国产精品久久久久影院| 999精品在线视频| 亚洲男人天堂网一区| 日韩高清综合在线| 黑人猛操日本美女一级片| 757午夜福利合集在线观看| 神马国产精品三级电影在线观看 | 国产精品野战在线观看 | 少妇的丰满在线观看| 韩国av一区二区三区四区| 999久久久国产精品视频| 久久精品影院6| 成人黄色视频免费在线看| 久久人人爽av亚洲精品天堂| 亚洲成人久久性| 欧美黄色片欧美黄色片| 中文欧美无线码| 亚洲国产中文字幕在线视频| 精品福利观看| 宅男免费午夜| 国产欧美日韩一区二区精品| 免费观看精品视频网站| 婷婷精品国产亚洲av在线| 午夜福利在线免费观看网站| 一进一出抽搐动态| 国产亚洲精品一区二区www| 国产男靠女视频免费网站| 91成年电影在线观看| 午夜老司机福利片| 咕卡用的链子| 在线观看一区二区三区激情| 亚洲九九香蕉| 午夜成年电影在线免费观看| 精品国产超薄肉色丝袜足j| 亚洲欧美日韩无卡精品| 久久欧美精品欧美久久欧美| 国产99白浆流出| 丰满饥渴人妻一区二区三| 国产99白浆流出| 亚洲av成人不卡在线观看播放网| 亚洲成人免费av在线播放| 波多野结衣av一区二区av| 老熟妇仑乱视频hdxx| 人人妻,人人澡人人爽秒播| 国产主播在线观看一区二区| 亚洲,欧美精品.| 女生性感内裤真人,穿戴方法视频| 国内毛片毛片毛片毛片毛片| 精品福利永久在线观看| 一个人免费在线观看的高清视频| 国产一区二区三区综合在线观看| 亚洲性夜色夜夜综合| 国产精品 国内视频| 后天国语完整版免费观看| 国产欧美日韩一区二区精品| 欧美精品亚洲一区二区| 老汉色av国产亚洲站长工具| 精品无人区乱码1区二区| 韩国av一区二区三区四区| 亚洲国产精品sss在线观看 | 免费在线观看日本一区| 欧美av亚洲av综合av国产av| 人成视频在线观看免费观看| 亚洲精品久久午夜乱码| 一级a爱片免费观看的视频| 69av精品久久久久久| 欧美中文综合在线视频| 国产视频一区二区在线看| 国产精品二区激情视频| 97超级碰碰碰精品色视频在线观看| 国产亚洲精品一区二区www| 国产亚洲精品第一综合不卡| 交换朋友夫妻互换小说| 人人澡人人妻人| 女人被躁到高潮嗷嗷叫费观| 满18在线观看网站| 看免费av毛片| 天天影视国产精品| 亚洲五月天丁香| 91大片在线观看| 超碰97精品在线观看| 国产精品九九99| 亚洲成a人片在线一区二区| 欧美性长视频在线观看| 天堂动漫精品| 精品无人区乱码1区二区| 欧美性长视频在线观看| 老汉色av国产亚洲站长工具| 欧美成人免费av一区二区三区| 欧美精品亚洲一区二区| 女性生殖器流出的白浆| 久久久久国产一级毛片高清牌| 老熟妇乱子伦视频在线观看| 久久久水蜜桃国产精品网| 免费不卡黄色视频| 欧美日韩瑟瑟在线播放| 精品乱码久久久久久99久播| 欧美日韩福利视频一区二区| 欧美日韩国产mv在线观看视频| 巨乳人妻的诱惑在线观看| 中亚洲国语对白在线视频| 高潮久久久久久久久久久不卡| 大型黄色视频在线免费观看| 啪啪无遮挡十八禁网站| 精品高清国产在线一区| 午夜福利一区二区在线看| 香蕉国产在线看| 在线国产一区二区在线| 1024香蕉在线观看| 十分钟在线观看高清视频www| 97人妻天天添夜夜摸| 男女床上黄色一级片免费看| 国产单亲对白刺激| 麻豆久久精品国产亚洲av | 亚洲专区国产一区二区| 久久久国产精品麻豆| 91麻豆av在线| 999久久久精品免费观看国产| 国产亚洲欧美在线一区二区| 侵犯人妻中文字幕一二三四区| 午夜福利一区二区在线看| 麻豆国产av国片精品| 欧美乱码精品一区二区三区| 少妇粗大呻吟视频| 日本五十路高清| 国产一区二区激情短视频| 两性夫妻黄色片| 男女高潮啪啪啪动态图| 国产精品国产高清国产av| 韩国精品一区二区三区| 成年人免费黄色播放视频| 久久99一区二区三区| 美女 人体艺术 gogo| 制服人妻中文乱码| 国产伦一二天堂av在线观看| 不卡一级毛片| 日韩欧美一区二区三区在线观看| 中文字幕人妻丝袜制服| 在线观看免费午夜福利视频| 久久精品亚洲熟妇少妇任你| 又黄又爽又免费观看的视频| 亚洲精品一区av在线观看| 欧美黑人欧美精品刺激| 巨乳人妻的诱惑在线观看| 亚洲五月婷婷丁香| www日本在线高清视频| 精品乱码久久久久久99久播| 精品久久蜜臀av无| 老熟妇仑乱视频hdxx| 黄频高清免费视频| 国产99久久九九免费精品| 涩涩av久久男人的天堂| 久久精品成人免费网站| 中文字幕av电影在线播放| 啦啦啦免费观看视频1| 又大又爽又粗| 色婷婷久久久亚洲欧美| 欧美日韩中文字幕国产精品一区二区三区 | 黄色 视频免费看| 亚洲九九香蕉| 国产精品一区二区免费欧美| 亚洲精品久久成人aⅴ小说| 成人黄色视频免费在线看| 欧美性长视频在线观看| 午夜福利免费观看在线| 正在播放国产对白刺激| 国产精品久久久av美女十八| 99热国产这里只有精品6| 亚洲一区中文字幕在线| 国产免费现黄频在线看| 亚洲在线自拍视频| 男女做爰动态图高潮gif福利片 | 久久久久精品国产欧美久久久| 大型黄色视频在线免费观看| 欧美精品一区二区免费开放| 在线十欧美十亚洲十日本专区| 欧美黑人欧美精品刺激| 男女高潮啪啪啪动态图| 啦啦啦免费观看视频1| 69精品国产乱码久久久| 亚洲五月色婷婷综合| 国产成人精品在线电影| 国产高清国产精品国产三级| 色在线成人网| 这个男人来自地球电影免费观看| 亚洲专区字幕在线| 女人爽到高潮嗷嗷叫在线视频| 国产一区在线观看成人免费| 亚洲国产欧美一区二区综合| 狂野欧美激情性xxxx| 免费一级毛片在线播放高清视频 | 999久久久精品免费观看国产| 国产真人三级小视频在线观看| 久久香蕉国产精品| 成年人黄色毛片网站| 亚洲欧美日韩高清在线视频| 日韩中文字幕欧美一区二区| 一进一出好大好爽视频| 免费搜索国产男女视频| 久久草成人影院| 香蕉久久夜色| 亚洲少妇的诱惑av| 俄罗斯特黄特色一大片| 女人被狂操c到高潮| 一本大道久久a久久精品| 亚洲国产精品999在线| 免费女性裸体啪啪无遮挡网站| 少妇的丰满在线观看| 成年版毛片免费区| 欧美激情高清一区二区三区| 看片在线看免费视频| 91麻豆精品激情在线观看国产 | 欧美午夜高清在线| 色综合欧美亚洲国产小说| 又紧又爽又黄一区二区| 男女下面插进去视频免费观看| 99国产精品99久久久久| 在线免费观看的www视频| 中出人妻视频一区二区| 久久人人97超碰香蕉20202| 日韩高清综合在线| 在线观看66精品国产| 免费观看人在逋| 日日爽夜夜爽网站| 亚洲成av片中文字幕在线观看| 美女扒开内裤让男人捅视频| 在线视频色国产色| 老司机午夜十八禁免费视频| 国产高清视频在线播放一区| 波多野结衣一区麻豆| 一二三四在线观看免费中文在| 99国产综合亚洲精品| 精品国内亚洲2022精品成人| 久久天堂一区二区三区四区| 国产精品久久电影中文字幕| 国产又色又爽无遮挡免费看| 成年女人毛片免费观看观看9| 好男人电影高清在线观看| 在线观看免费日韩欧美大片| 黑人巨大精品欧美一区二区mp4| 51午夜福利影视在线观看| 国产成人精品久久二区二区免费| 亚洲熟妇熟女久久| www.熟女人妻精品国产| 少妇被粗大的猛进出69影院| 免费久久久久久久精品成人欧美视频| 久久久国产欧美日韩av| 一区在线观看完整版| 亚洲人成网站在线播放欧美日韩| 91av网站免费观看| 一本综合久久免费| 久久人妻福利社区极品人妻图片| 国产亚洲精品综合一区在线观看 | 久久精品亚洲熟妇少妇任你| 国产伦人伦偷精品视频| 又黄又粗又硬又大视频| 久久中文字幕一级| 亚洲欧美一区二区三区久久| 色综合站精品国产| 看片在线看免费视频| 国产精品二区激情视频| 50天的宝宝边吃奶边哭怎么回事| 国产av一区二区精品久久| 精品午夜福利视频在线观看一区| 天天影视国产精品| 桃色一区二区三区在线观看| 亚洲精品在线观看二区| aaaaa片日本免费| 亚洲国产欧美日韩在线播放| 成人黄色视频免费在线看| 国产视频一区二区在线看| 啪啪无遮挡十八禁网站| 国产精品99久久99久久久不卡| 亚洲精品中文字幕一二三四区| 91在线观看av| 久久中文字幕一级| 国产免费男女视频| 一夜夜www| 国产熟女xx| 淫秽高清视频在线观看| 亚洲av成人一区二区三| 国产成人精品在线电影| 国产成人精品久久二区二区免费| 亚洲男人天堂网一区| 少妇的丰满在线观看| av免费在线观看网站| 欧美日本中文国产一区发布| 操出白浆在线播放| 老司机亚洲免费影院| 国产av一区二区精品久久| 国产极品粉嫩免费观看在线| netflix在线观看网站| 一区福利在线观看| 成人手机av| 91在线观看av| 他把我摸到了高潮在线观看| 午夜老司机福利片| 一级黄色大片毛片| 国产一区二区激情短视频| 成人国语在线视频| 欧美成人午夜精品| 十八禁网站免费在线| 国产成人欧美在线观看| 国产亚洲av高清不卡| 欧美黑人欧美精品刺激| 国产成人av教育| 亚洲午夜精品一区,二区,三区| 村上凉子中文字幕在线| 亚洲欧美一区二区三区黑人| 国产成人啪精品午夜网站| 欧美成人午夜精品| 亚洲av成人av| 制服诱惑二区| 91大片在线观看| 自线自在国产av| 亚洲av熟女| 丰满人妻熟妇乱又伦精品不卡| 欧美日韩瑟瑟在线播放| 欧美黑人欧美精品刺激| 日韩一卡2卡3卡4卡2021年| 国产免费男女视频| 精品国产一区二区三区四区第35| 十八禁人妻一区二区| 韩国av一区二区三区四区| 欧美av亚洲av综合av国产av| 视频在线观看一区二区三区| 老司机亚洲免费影院| 黄频高清免费视频| 欧美日韩av久久| 亚洲国产欧美一区二区综合| 中文亚洲av片在线观看爽| 一边摸一边抽搐一进一小说| 中文亚洲av片在线观看爽| 一区二区三区精品91| 在线观看免费视频日本深夜| 国产色视频综合| 国产成人欧美| 国产麻豆69| 亚洲av成人不卡在线观看播放网| 在线av久久热| 精品一品国产午夜福利视频| 高清在线国产一区| 岛国视频午夜一区免费看| 淫妇啪啪啪对白视频| 欧美黄色片欧美黄色片| 国产精品野战在线观看 | 在线av久久热| 丁香欧美五月| 国产97色在线日韩免费| 一级a爱片免费观看的视频| 久久久国产一区二区| 国产一区在线观看成人免费| 在线播放国产精品三级| 99国产精品免费福利视频| 高潮久久久久久久久久久不卡| 在线天堂中文资源库| 精品国内亚洲2022精品成人| 嫩草影院精品99| 香蕉久久夜色| 免费人成视频x8x8入口观看| 亚洲三区欧美一区| 在线观看免费午夜福利视频| 亚洲成人免费av在线播放| 久久久国产成人精品二区 | 人人澡人人妻人| 日韩三级视频一区二区三区| 女人爽到高潮嗷嗷叫在线视频| 精品免费久久久久久久清纯| 久久国产精品影院| 男人操女人黄网站| 99国产极品粉嫩在线观看| 狂野欧美激情性xxxx| 在线观看免费视频网站a站| 中出人妻视频一区二区| 丰满的人妻完整版| 亚洲欧美日韩高清在线视频| 亚洲人成77777在线视频| 搡老熟女国产l中国老女人| 精品免费久久久久久久清纯| 久久国产精品影院| 在线观看日韩欧美| 在线视频色国产色| 正在播放国产对白刺激| 精品一区二区三区视频在线观看免费 | 久久久国产成人精品二区 | 夜夜躁狠狠躁天天躁| 日韩三级视频一区二区三区| 亚洲国产欧美网| 亚洲欧美一区二区三区黑人| 国产精品98久久久久久宅男小说| 亚洲色图 男人天堂 中文字幕| 精品国产亚洲在线| 亚洲人成电影观看| 亚洲精品一卡2卡三卡4卡5卡| 欧美日韩乱码在线| 亚洲精品av麻豆狂野| 天天影视国产精品| 国产成人一区二区三区免费视频网站| 欧美日韩瑟瑟在线播放| 黄色视频不卡| 老司机福利观看| 999久久久精品免费观看国产| 国产成人精品无人区| 日日摸夜夜添夜夜添小说| 国产精品久久久人人做人人爽| 桃色一区二区三区在线观看| 女同久久另类99精品国产91| 精品久久久久久久毛片微露脸| 国产高清国产精品国产三级| 亚洲国产精品999在线| 99香蕉大伊视频| 欧美国产精品va在线观看不卡| 国产一区二区三区综合在线观看| 精品久久久久久成人av| 99国产极品粉嫩在线观看| 69精品国产乱码久久久| 国产亚洲精品久久久久久毛片| 久久九九热精品免费| 97碰自拍视频| 亚洲国产中文字幕在线视频| 免费高清视频大片| 中文字幕精品免费在线观看视频| 视频在线观看一区二区三区| 亚洲专区中文字幕在线| 中出人妻视频一区二区| 亚洲av美国av| 丝袜美足系列| 亚洲国产精品一区二区三区在线| 丰满饥渴人妻一区二区三| 人人妻人人澡人人看| 久久国产精品男人的天堂亚洲| 国产aⅴ精品一区二区三区波| 日韩三级视频一区二区三区| 一本大道久久a久久精品| 日韩人妻精品一区2区三区| 悠悠久久av| 操美女的视频在线观看| 免费av毛片视频| 国产99久久九九免费精品| 少妇裸体淫交视频免费看高清 | 狠狠狠狠99中文字幕| 一级a爱视频在线免费观看| 国产成人影院久久av| 嫩草影视91久久| 成人亚洲精品av一区二区 | 久久精品亚洲精品国产色婷小说| 一级,二级,三级黄色视频| 女人被躁到高潮嗷嗷叫费观| 亚洲第一欧美日韩一区二区三区| 午夜福利,免费看| 咕卡用的链子| 国产精品久久视频播放| 欧美+亚洲+日韩+国产| 欧美日韩亚洲国产一区二区在线观看| 国产精品免费一区二区三区在线| 在线国产一区二区在线| 老鸭窝网址在线观看| 国产成人欧美| 老司机福利观看| 免费高清视频大片| 老司机亚洲免费影院| 欧美精品一区二区免费开放| 亚洲成人免费av在线播放| 最新美女视频免费是黄的| 精品久久久久久久毛片微露脸| av免费在线观看网站| 一进一出抽搐动态| 男女做爰动态图高潮gif福利片 | 欧美日韩av久久| 精品熟女少妇八av免费久了| 男女下面进入的视频免费午夜 | 国产成人av教育| 精品一品国产午夜福利视频| 久久 成人 亚洲| av网站免费在线观看视频| 欧美一区二区精品小视频在线| 欧美成人免费av一区二区三区| 一二三四社区在线视频社区8| 午夜福利影视在线免费观看| www.www免费av| 亚洲精品在线观看二区| 国产精品一区二区在线不卡| 大陆偷拍与自拍| 国产亚洲精品久久久久久毛片| 操美女的视频在线观看| 91麻豆精品激情在线观看国产 | 亚洲一区二区三区欧美精品| 亚洲全国av大片| 亚洲成国产人片在线观看| 人人妻人人爽人人添夜夜欢视频| 免费在线观看视频国产中文字幕亚洲| 香蕉国产在线看| 丁香欧美五月| 99精品在免费线老司机午夜| av网站免费在线观看视频| 自线自在国产av| 伦理电影免费视频| 女生性感内裤真人,穿戴方法视频| 色老头精品视频在线观看| 欧美日韩福利视频一区二区| 一区福利在线观看| 午夜日韩欧美国产| 久久精品成人免费网站| 色在线成人网| 怎么达到女性高潮| 最近最新中文字幕大全免费视频| 三上悠亚av全集在线观看| 精品久久久久久成人av| 午夜福利免费观看在线| 国产亚洲精品久久久久久毛片| 欧美黄色淫秽网站| 长腿黑丝高跟| 又黄又爽又免费观看的视频| 黄色片一级片一级黄色片| 两个人免费观看高清视频| 看黄色毛片网站| 嫁个100分男人电影在线观看| 久久欧美精品欧美久久欧美| 久久国产精品人妻蜜桃| 亚洲精品中文字幕一二三四区| 亚洲成人久久性| 多毛熟女@视频| 大陆偷拍与自拍| 超碰97精品在线观看| 欧美另类亚洲清纯唯美| 亚洲av电影在线进入| 精品国产亚洲在线| 老司机福利观看| 久久久久久大精品| 中文亚洲av片在线观看爽| 中文字幕色久视频| 国产三级黄色录像| 一边摸一边做爽爽视频免费| 欧美丝袜亚洲另类 | 夜夜夜夜夜久久久久| 国产精品二区激情视频| 9热在线视频观看99| 色婷婷av一区二区三区视频| 日本精品一区二区三区蜜桃| 手机成人av网站| 日本撒尿小便嘘嘘汇集6| 天堂动漫精品| 亚洲专区字幕在线| 国产成人影院久久av| 亚洲精品在线美女| 精品久久久久久电影网| 在线观看一区二区三区| xxx96com| 国产精品日韩av在线免费观看 | 久久久久久久久免费视频了| 色综合站精品国产| 老司机深夜福利视频在线观看| 国产精品 国内视频| 丝袜在线中文字幕| 免费不卡黄色视频| 国产av一区二区精品久久| 国产激情欧美一区二区| 嫩草影院精品99| 一级a爱视频在线免费观看| 久久九九热精品免费| www.自偷自拍.com|