Insemination with frozen dog semen based on a new insemination technique
ABSTRACT Inhalt: (Insemination mit Gefriersperma beim Hund mit Hilfe einer neuen Besamungstechnik).In diesem Versuch sind 11 Hündinnen mit tiefgefrorenem Sperma besamt worden.Die spermienreiche Ejakulatsfraktion wurde unmittelbar nach der Samenentnahme mit einer Tris-Fructose-Zitronensüure-Lösung, die 8 Vol. % Glyzerin und 20 Vol. % Eidotter enthielt, etwa 1: 4 verdünnt. Wührend einer 3stündigen Equilibrierungszeit wurde der Samen auf + 5 °C heruntergekühlt, and in PVC-Röhrchen im Stickstoffdampf eingefroren.Die Inseminationsdosen, die ungefähr 150 × 106 Spermien enthielten und 3 Wochen bis 1 1/2 Jahre gelagert waren, wurden unmittelbar nach dem Auftauen für 6,5 Sek. im Wasserbad von + 75 °C, durch den Cervicalkanal intrauterin deponiert. Es wurde meistens zweimal mit etwa 48stündigen Intervallen inseminiert. Von den 11 Hündinnen konzipierten 10 und brachten von ein bis sieben Junge. Die höchste Zahl der lebendigen Welpen in einem Wurf war sechs.Contents:Insemination with frozen dog semen was performed in a small trial including 11 bitches. The semen was diluted about 1: 4 with Tris-fructose-citric acid extender containing 8 % (v/v) glycerol and 20 % (v/v) egg yolk, equilibrated for 3 hrs and frozen in P.V.C.-straws by use of N2-vapour. The insemination doses being stored in liquid N2 from 3 weeks to 1 1/2 year and containing about 150 × 106 spermätozoa, was deposited in the uterus via the cervical canal immediately after thawing at 75 °C for 6,5 sec. By this procedure conception was obtained in 10 of the 11 bitches. The litter size ranged from one to seven puppies, the highest number of living puppies being six.
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- "The spermatozoa were loaded into a 0.25 mL-straw and then frozen by placing the straw horizontally 4 cm above liquid nitrogen vapors for 10 min. The straw was finally immersed in the liquid nitrogen (Andersen, 1975).. Thawing procedure was performed by immersing the straw in warm water (37C) for 15 s. The frozen-thawed semen was then released into a pre-warmed thawing medium (37C). "
ABSTRACT: Cryopreservation of epididymal sperm is an effective technique to preserve genetic materials of domestic cats and wild felids when they unexpectedly die. However, this technique inevitably causes detrimental changes of cryopreserved-thawed spermatozoa, for example, by physical damage and excessive oxidative stress. L-carnitine is an antioxidant that has been used to improve sperm motility in humans and domestic animals. This study aimed to investigate the effects of L-carnitine on cat epididymal sperm quality following cryopreservation and thawing. After routine castration, cauda epididymides were collected from 60 cat testes. The epididymal spermatozoa from 3 cauda epididymides were pooled as 1 replicate. Spermatozoa samples (16 replicates) were examined for spermatozoa quality and then randomly divided into 4 groups: 0 mM L-carnitine (control), 12.5 mM, 25 mM and 50 mM L-carnitine. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, plasma membrane integrity, DNA integrity and acrosome integrity were evaluated. The 25 mM L-carnitine significantly improved sperm motility compared with a control group (p<0.05), although this was not significantly different among other concentrations. In conclusion, supplementation of 25 mM L-carnitine in freezing extender improves cauda epididymal spermatozoa motility. The effects of L-carnitine on the levels of oxidative stress during freezing and thawing remains to be examined.Asian Australasian Journal of Animal Sciences 06/2014; 27(6):791-6. DOI:10.5713/ajas.2013.13565 · 0.56 Impact Factor
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- "In the last 25 years, a lot of research has been conducted on different aspects of canine reproduction, and AI technology has been well developed. Especially the technique of intrauterine insemination using the Scandinavian catheter (Andersen, 1975; Linde-Forsberg, 2001) without any surgical intervention increased the success rate of AI in the dog (Linde-Forsberg and Forsberg, 1989; Rota et al., 1999, Linde-Forsberg 1995; 2001). Today AI in the dog is a common procedure in veterinary clinics. "
ABSTRACT: This study investigated whether the immotility induced by the CLONE chilled semen extender prolongs the lifespan of dog spermatozoa stored at 5 degrees C, compared with a Tris-egg yolk-glucose (TG) extender, which maintains motility. Pooled semen was split in four aliquots, centrifuged, and the four sperm pellets mixed with TG extender; with the CLONE chilled semen (CL) extender; with TG extender mixed with an activator (TG+A(TG)); or with the CLONE extender mixed with the CLONE activator (CL+A(CL)). Samples were stored at 5 degrees C for 23 days and examined 12 times for sperm motility, plasma membrane and acrosome integrity, glucose consumption, and DNA fragmentation index (DFI). The experiment was performed in triplicate. Glucose consumption was not significantly different between extenders until the period 15-23 days, when it was higher in CL and CL+A(CL) than in TG (P=0.0055) and TG+A(TG) (P=0.0010). No breakdown of DNA chromatin (P>0.05) occurred until day 14. Spermatozoa preserved in TG or TG+A(TG) showed better values for all the different parameters throughout the experiment compared with sperm subjected to CL or CL+A(CL). In conclusion, the immotility induced by the CLONE chilled semen extender during long-term cold storage at 5 degrees C did not prolong the lifespan of spermatozoa compared with the lifespan following storage in Tris-egg yolk-glucose. In addition, our results indicate that good quality dog semen may possibly be stored for up to 14 days in TG extender at 5 degrees C, with retained fertilizing capacity. In vivo studies should, however, be performed to further support this conclusion.Theriogenology 11/2007; 68(6):920-33. DOI:10.1016/j.theriogenology.2007.07.006 · 1.85 Impact Factor
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- "Since the work of Anderson in 1975 , it was accepted that frozen-thawed dog sperm should be inseminated into the uterus in order to obtain good fertility. Uterine insemination poses practical problems because it either requires one to pass a pipette through the cervix, which is sometimes impossible to achieve  and , or expensive fiber-optic equipment with which to catheterize the cervix under visual control , or surgical or laparoscopic exposure of the uterus  and . "
ABSTRACT: Although dog prostatic fluid decreases the longevity of ejaculated dog spermatozoa, it also increases their rate of motility and their fertility after vaginal insemination, as well as the fertility of epididymal spermatozoa after uterine insemination. These findings indicate a need to further characterize the effects of prostatic fluid on dog spermatozoa. This study was done to determine the effects (P<0.05) of homologous prostatic fluid added prior to cooling, after thawing, or at both times to epididymal spermatozoa from 21 dogs. The effects of two extenders were also determined. The one extender was Biladyl(*) with Equex STM paste(**) (BilEq) and the other Andromed(*) (Minitüb, Tiefenbach, Germany (*); Nova Chemical Sales, Scituate, MA, USA (**)). The response variables were percentage progressively motile spermatozoa (Prog) and morphology after thawing. Prog was measured at various times until 8h after extension (unfrozen spermatozoa) or until 2h after thawing. Prog after thawing was higher with BilEq than Andromed, when no prostatic fluid was added prior to cooling, and when prostatic fluid was added after thawing. BilEq resulted in a higher mean percentage of spermatozoa with bent principle pieces than Andromed and the addition of prostatic fluid prior to cooling resulted in lower mean percentages of cytoplasmic droplets and bent principle pieces than when none was added. The optimal combination was BilEq with prostatic fluid added prior to cooling (in order to inhibit the development of bent principle pieces) and after thawing (to achieve higher motility until 1h after thawing). This study shows that BilEq is more suitable for the freezing of epididymal spermatozoa than Andromed and that prostatic fluid improves the freezability and post-thaw longevity of epididymal spermatozoa frozen in BilEq.Theriogenology 02/2007; 67(2):264-75. DOI:10.1016/j.theriogenology.2006.07.018 · 1.85 Impact Factor