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N];Kfa+g9s$["B;͒>vW^Z>T}f/;cg4{:4{Qbp{U?p Y?Zn?sn?>2!_sj?4~~x]?|E1qഞ3xc3&@Ke6JȬW=s营rGt<7Ou>̧2rd:3Og3si͑7rLwEџ$ym6Uֶft?k_)D?#5Ǹ7\=$[dS| _E&T=%OiYENU3p߀>Ä:83g|7^k8 X+9͌7aO0egSt&?1]NfiiFi%{LVEyM}E1}VdttrNC1O˘O8|\\=38U}l*d5+?B#rWO0_[Nm[T;V4޹{ 8 1/gY2X^lL(   $Paintbrush Picture  PBrush0$Paintbrush Picture$Paintbrush Picture  PBrush0$Paintbrush Picturev Chart Excel.Chart.50 Chart/ 0DArialNew <(0(z[ 0 "DMonotype Sorts(0(z[ 0  DTimes New Roman(0(z[ 0  B . @n?" dd@  @@`` x4<      ?b$qԨ16=Jb$p+#8ABHe\b=J2$P,ЎܰO fAA1?0 aCP@3 ʚ;ʚ;g4KdKd@z[ 0ppp@ <4ddddl 0x<80___PPT10 ppf[COMPARATIVE STUDY OF DIFFERENT CRYOPRESERVATION METHODS FOR DEEP FREEZING OF CANINE SEMEN. \\$ \1Szsz F, 2Gbor G, 1Solti L. 1Dept. Obstet. and Reprod. Univ. Vet. Sci. Budapest, Hungary, 2Dept. Cattle Breeding, Res. Inst. Anim. Breed. Herceghalom, Hungary.    ?E   INTRODUCTION   In the process of fertilization, following capacitation, spermatozoa undergo the acrosome reaction (AR) during penetration of the zona pellucida. AR involves the breakdown and fusion of the plasma membrane and outer acrosomal membrane. During the freezing/thawing procedures the acrosomal membrane can be damaged and it can cause lower fertilizing ability of sperm. Lectins (FITC-PNA) have been widely used to visualize the acrosomal status and membrane damage of spermatozoa.   OBJECTIVES   yComparison of different extenders different freezing rates on the post-thaw motility and acrosome integrity of sperm.*/<z z MATERIAL and METHODS I. Second fraction ejaculates were collected from 3 bull-mastiff dogs. Samples with good sperm motility (80%) were mixed, diluted upto a sperm concentration (100x106/ml) then were frozen, used three different freezing rates and extenders. Regular bull semen freezing rate (1) - Triladyl extender (MINITB, Germany)(A) Canine protocol (CP) by Farstad (2)  tris-fructose-citric acid extender (B) CP by University of Utrecht (3)- tris-fructose-citric acid extender with glycin (C).  , %$'7  MATERIAL and METHODS II.  Each freezing protocols and extenders were used and compared with each other. Ten straws of each combinations were thawed (70 C, 8 sec). Post-thawing motility was estimated by light microscopy. Acrosome integrity was examined by FITC-PNA fluorescence staining.   MATERIAL and METHODS III.+ FITC-PNA fluorescence staining: After thawing, the samples were washed, fixed (2% paraformaldehyde), and labeled with FITC-PNA. Of each slide, 200 sperm cells were assessed under an epifluorescent microscope (x1,250). 0 ~\  PICTURE 1.     ?(1) acrosome intact(AInt)cells: there is no green fluorescence at the acrosomal region, only a very faint green fluorescence surrounding the plasma membrane. (2) acrosome damaged (AD) cells: green fluorescence at the acrosomal region. (3) acrosome reacted (AR) cells: green fluorescence only at the equatorial segment.d@PP2P5 @RESULTS AND DISCUSSION NThe post-thawing motility rate were between 30-50 %, depending on the freezing protocol and the extender. Significant differences were found between the percentage of spermatozoa with intact acrosome in the different semen samples (P<0.001). The best motility and integrity rates (P>0.05) were found in 2/A, 2/B and 3/C combinations.OO OaAverage rate of acrosome intact sperm and motility in each of freezing rate/extender combinationsbb  b  CONCLUSION    It was concluded that the freezing rates have a higher impact on the post-thawing semen quality than the extender. The regular bull semen freezing rate is not useful for the canine semen that may due to the slower temperature changes during the freezing protocol. 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