Fertility of undiluted ram epididymal spermatozoa stored for several days at 4°C

Abstract

In vitro preservation of the male gamete is a challenge in the development of artificial insemination techniques for domestic animals. Specific strategies and diluents have been developed for the preservation of the fertilizing ability of the semen for each species. However, the epididymal medium has been demonstrated to be the best sperm environment to maintain sperm viability over several days and weeks for mammals. The aims of this study were to evaluate the motility and in vivo fertility of ram epididymal spermatozoa when the semen was stored for up to 4 days at 4°C undiluted in epididymal plasma. The study was undertaken with two ovine breeds (Ile de France and Corriedale). The motility of epididymal spermatozoa was better preserved in the undiluted epididymal fluid than when epididymal spermatozoa were diluted in classic ovine extender such as skim milk. During storage, the decrease in the percentage of motile sperm was lower if the epididymal spermatozoa were collected immediately after epididymal sampling than 24 h after castration or animal death. The fertility obtained after cryopreservation of the stored sperm and subsequent intrauterine insemination ranged from 55% to 24% following 24 to 96-h sperm storage. There was a linear regression relationship between fertility and the number of motile sperm inseminated for both breeds. These results show that it is possible to keep epididymal sperm motile and fertile for several days without dilution. Such a method of sperm preservation could be a final possibility for animals of high genetic value or for endangered species when the collection of semen before death of the animal is not possible.

Development of Porcine Accessory Sex Glands

“…2 Sections were incubated with the indicated dilution of the primary antibody in PBS, PBS containing 0.05% Tween (PBST), Tris-buffered saline (TBS), or TBS containing 20% normal rabbit serum and 50 mg BSA/mL (TBST). 3 NovaRed ® (SK4800), AEC (3-amino-9-ethylcarabazole; SK4200), and ImmPACT Vector Red (SK5105) were all obtained from Vector Laboratories, Burlingame, CA, USA, and DAB (3,3-diaminobenzidine; K3468) was obtained from DAKO Corporation, Carpinteria, CA, USA.…”

“…Accessory sex glands have long been recognized as steroid-responsive tissues that can become neoplastic in later life, causing significant mortality in men [1,2]. Secretions from these glands are not essential to fertilization since epididymal sperm are capable of fertilization in vivo following artificial insemination in multiple species [3][4][5][6][7][8][9][10][11][12]. However, the infertility in vivo of some rodent ejaculates following the removal of individual accessory sex glands indicates a functional role for these glands [13][14][15].…”

Effect of Toxoplasma gondii on Ram Sperm Quality after Experimental Infection

“…A possible increased susceptibility of infected animals' spermatozoa membranes under handling and thermal processes led to lower viability compared to motility values. Semen morphology and the relationship with fertility has also been confirmed as an important indicator of semen fertilizing capacity in different species [16,47,48]. Despite sulphadimidine treatment, the infected groups showed higher rates of head, tail, and total morphological abnormalities compared to the control group.…”

“…BCF is the number of times the sperm head crosses the direction of movement, developing another flagellar wave. This result, alone, cannot be appreciated as high value, because among CASA semen evaluated parameters, assessment in combination with progressive motility, ALH, BCF, and VSL are much more correlated with in vivo fertility than the estimation of single kinetic parameter [47]. Additionally, according to Herrara et al [55] in vitro fertilization (IVF) is significantly correlated to progressive motility but not to BCF.…”

“…Both testes and epididymides contained in scrotal sacs were immediately removed after slaughtering, placed into an isothermal box (15 • C), and transferred to the laboratory in about 2 h. At room temperature, about 20 • C, each epididymal cauda was revealed from the scrotum, detached from the respective testis, and weighed on a high precision scale for the evaluation of sperm concentration. The epididymal sperm was collected through the proximal end of the severed epididymis by infusion of 5 mL Phosphate Buffered Saline (PBS) under pressure from the aperture of the deferent duct [47,68]. The 10 mL blend of each pair of epididymides were centrifuged (1500 rpm; 10 min; 20 • C), and the sediment was diluted to a 1:2 ratio with Ovixcell ® ram semen extender (IMV Technologies, L'Aigle, France) and preserved in a water bath (37 • C) until the end of the evaluation.…”