The effect of the breeding season, cryopreservation and physiological extender on selected sperm and semen parameters of four ferret species: implications for captive breeding in the endangered black-footed ferret

2009 ◽  
Vol 21 (2) ◽  
pp. 351 ◽  
Author(s):  
G. van der Horst ◽  
R. M. Kitchin ◽  
M. van der Horst ◽  
R. W. Atherton

In the present investigation, comparative baseline information on selected sperm characteristics of ejaculate spermatozoa of the domestic (Mustela putorius furo), fitch (Mustela sp.) and black-footed ferrets (Mustela nigripes) and the Siberian polecat (Mustela eversmanni) are presented. The main emphasis was to establish differences and similarities among these species in relation to semen and sperm quality during the breeding season, in cryopreservation success and in supporting sperm motility in different extenders or physiological media. The results confirm that most sperm morphology abnormalities were evident during the beginning of the breeding cycle in all four species. No significant interspecies differences were apparent in the sperm attributes examined, for all sampling months during the breeding season. Moreover, all species exhibited comparable patterns of reproductive seasonality. Cryopreservation suppressed sperm characteristics equally in all species studied. Ejaculate spermatozoa of closely related ferret species shared many similar motion characteristics using computer-aided sperm motility analysis. These results suggest that the basic sperm physiology of the ferret species under examination is very similar. Disparate to the interspecies comparisons, there were significant differences for most sperm motion parameters when spermatozoa of any of the ferrets were compared in different extenders. Assisted reproductive technologies developed for use in domestic ferret, fitch ferret or Siberian polecat may be successfully applied to captive breeding of the black-footed ferret using semen during any of the functional breeding months.

Biology ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 33
Author(s):  
Christine Green ◽  
Jessica P. Rickard ◽  
Simon P. de Graaf ◽  
Angela J. Crean

Males can adjust sperm motility instantaneously in response to the perceived risk of sperm competition. The speed of this response suggests that sperm motility is regulated by changes in seminal plasma rather than changes in the sperm cells themselves. Hence, here we test whether inter-ejaculate variation in seminal plasma can be used to alter sperm quality prior to use in assisted reproductive technologies. We supplemented fresh ejaculates of Merino rams with seminal plasma collected from previous ‘donor’ ejaculates to test whether changes in sperm kinetics were related to the relative quality of donor to focal ejaculates. We found a positive relationship between the change in sperm traits before and after supplementation, and the difference in sperm traits between the donor and focal ejaculate. Hence, sperm motility can be either increased or decreased through the addition of seminal plasma from a superior or inferior ejaculate, respectively. This positive relationship held true even when seminal plasma was added from a previous ejaculate of the same ram, although the slope of the relationship depended on the identity of both the donor and receiver ram. These findings indicate that seminal plasma plays a key role in the control and regulation of sperm kinetics, and that sperm kinetic traits can be transferred from one ejaculate to another through seminal plasma supplementation.


2006 ◽  
Vol 18 (2) ◽  
pp. 166
Author(s):  
G. Wirtu ◽  
C. E. Pope ◽  
A. Cole ◽  
R. A. Godke ◽  
D. L. Paccamonti ◽  
...  

As an integral aspect of our program to develop assisted reproductive technologies in tragelaphine antelopes, we have been evaluating the effects of various extenders on cryosurvival of spermatozoa. Semen was collected by electroejaculation (EEJ) from an eland (Taurotragus oryx, n = 14 EEJs) and a bongo (Tragelaphus euryceros, n = 7 EEJs) male during sedation and standing restraint in a hydraulic chute. Epididymal spermatozoa were collected from the proximal vas deferens, and distal epididymides recovered from four common eland bulls after elective castration at the Hattiesburg Zoo, Mississippi (age = 46 months) and the San Diego Wild Animal Park, California (ages = 18, 20, and 23 months). Testes from a bongo (age = 6.5 yr) and a greater kudu bull (Tragelaphus strepsiceros, age = 7.5 yr) at the Audubon Nature Institute were recovered and processed immediately postmortem. Kudu, bongo, and the Hattiesburg eland testes were transported at ambient temperature whereas the testes from San Diego elands were shipped overnight in a cold-pack container. Sperm processing was done at room temperature (RT = 23°C) in HEPES-buffered Tyrode's solution. Four extenders containing 7% glycerol were evaluated: Biladyl®, TEST Yolk buffer (TYB), Beltsville extender (BF5F), and skim milk. For freezing, sperm samples were initially extended and gradually cooled to 4°C before sequential addition of extender containing glycerol at 4°C; the procedure was then modified by addition of the glycerol fraction at RT before the sample was cooled to 4°C. Spermatozoa were vapor-frozen in 0.5 mL straws placed 5 cm above liquid nitrogen before storage at −196°C. Straws were thawed in a water bath (38°C, 30 s) to evaluate cryosurvival. All eland and bongo electroejaculation procedures produced spermatozoa, although sperm quality varied. Ejaculate volume averaged 5.4 ± 1.2 mL and 3.7 ± 1.1 mL in the bongo and the eland, respectively. Sperm motility at collection, after cooling, and after freezing in the different extenders is presented in Table 1. No immediate decline in sperm motility was observed after adding glycerol to samples, whether at 23°C or 4°C. Bongo spermatozoa lost more motility during freezing than during cooling (15.0 ± 4.4 vs. 5.8 ± 2.0%; P = 0.006); whereas, in eland, motility loss during cooling and freezing was similar (25.3 ± 16.2 vs. 21.1 ± 7.7; P = 0.44). The present results indicate that cooling and freezing tolerance of spermatozoa in tragelaphine antelopes is influenced by species, extender type, and temperature at which a cryoprotectant is added. Table 1. Cryosurvival of ejaculated or epididymal spermatozoa of three antelope species in different extenders


2021 ◽  
Vol 63 (1) ◽  
Author(s):  
Marta Neila-Montero ◽  
Marta F. Riesco ◽  
Mercedes Alvarez ◽  
Rafael Montes-Garrido ◽  
Juan Carlos Boixo ◽  
...  

Abstract Background Centrifugation is routinely employed in handling the ejaculates of some species, but it is not part of the commonly used protocols in ram. However, the development and implementation of new assisted reproductive technologies, alternative preservation models based on washing sperm from a cellular ageing-accelerating substance such as the seminal plasma, and basic studies in spermatology is associated with the use of centrifugation. This requires a specific evaluation of the centrifugation protocols considering the species-specific relationship with the potential damage produced by this procedure. No previous studies have determined the effect of different centrifugation forces on ram sperm. Therefore, we aimed to assess the performance of three centrifugal forces (600×g, 3000×g, and 6000×g for 10 min at room temperature) and their effects on ram sperm motility and functionality. Results Sperm motility and functionality parameters were assessed at 0 h and after 2 h of incubation at 37 °C. As expected, a higher cell packaging degree was obtained at high centrifugation forces (P ≤ 0.0001). Cell packaging was unstable at all centrifugal forces. Thus, there was a high cell resuspension rate after less than 2 min. Regarding sperm quality, there was a change in movement pattern of 3000×g and 6000×g centrifuged sperm after 2 h of incubation at 37 °C, characterized by an increase in rapid progressive motility, linearity, straightness, and beat frequency, and a decrease in medium progressive motility, curvilinear velocity, path velocity, and head lateral amplitude. Non-significant differences were obtained among the different treatments concerning the total viability. However, we observed a significant increase (P ≤ 0.05) in the percentage of viable apoptotic sperm in the samples centrifuged at 6000×g at 0 h. Conclusions Centrifugal forces equal to or greater than 3000×g induced some deleterious effects in ram sperm quality, and lower forces did not provide a successful cell packaging degree.


Infertility rates in India becoming increased in last decade principally due to the urbanization conditions and the lifestyle habits. It is giving alarm by continuously reporting the progress in incident cases of infertility amongst the young Indian adults of both male and female population. Among the various Assisted Reproductive Technologies (ART) available today in the treatment of infertility, In Vitro Fertilization (IVF) is found to be the most applicable treatment method of choice. This involves the administration of different hormones and drugs to treat infertility. In the present scenario technically IVF treatment process is tedious, laborious, high cost and most importantly success rates reported to be very low (20-30%). The prediction of IVF success rates is becoming an important scientific knowledge and practice, which helps both the doctor and the candidate couple to know about the conditions hence to take the right decision. The accurate prediction of the IVF success rate is really a challenging task in obstetrics and gynecology medicine. The success rates of the IVF depends on the various factors such as Intrinsic factors i.e, Genetic predisposition, Age, Body mass Index, Hormonal balance, Embryo viability, Sperm quality, Endometriosis and overall patient’s response level of the candidate couple and the Extrinsic factors such as Medical equipment technology, Treatment methods, Personal experiences of clinicians and embryologists, Process time, Stress due to the lifestyle etc.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
O Somova ◽  
H Ivanova ◽  
N Sotnyk ◽  
K Kovalenko ◽  
I Feskova

Abstract Study question To evaluate the effect of platelet-rich plasma (PRP) testicular injections on spermogram parameters of men with severe oligoasthenoteratozoospermia (OAT). Summary answer The PRP testicular injections have beneficial effects on spermatogenesis and enhance sperm concentration and motility in infertile men with OAT. What is known already The use of PRP therapy in assisted reproductive technologies is debatable. Despite the recent evidence of its positive effects in promoting endometrial and follicular growth, data from clinical studies are limited. There are only a few papers on the effectiveness of PRP therapy in the treatment of male infertility and sexual dysfunction. In more detail, the influence of PRP on spermatogenesis was carried out only on experimental animals. Although the mechanisms of its action have not yet been clarified, it is assumed that PRP, containing many biologically active molecules, realizes its effect through the tissue regeneration and cell proliferation. Study design, size, duration This prospective study included 68 men (34.6±5.2) years old with severe OAT (≤4 million/ml, motility ≤30%, normal sperm morphology ≤1%) receiving hormonal and antioxidant (AO) therapy during 6 months before in vitro fertilization cycles. 33 of them were injected once with autologous PRP (0.5 ml in each testicle). Spermogram and testosterone level were analyzed before the treatment and in 3, 4 and 6 months after it. Participants/materials, setting, methods: Sperm concentration, motility and morphology in ejaculate of 33 men of PRP group were compared with those in the group of 35 men without PRP within 6 months of starting the treatment. Total and free testosterone level were measured in blood serum. PRP was prepared by centrifuging the patient’s own blood in the anticoagulant-containing tubes. The final concentration of platelets in the obtained sample was 950.000 – 1.250 000 cells in 1 ml. Main results and the role of chance 4 months after the PRP injection, sperm concentration and motility increased in 18 of 33 men of the PRP group compared with the baseline (before the treatment) – 4.2 (1.0; 6.9) vs 1.4 (0.1; 3.4) mln/ml (p < 0.05) and 36.7 (30.6; 45.8) vs 17.7 (6.7; 28.2)% respectively (p < 0.05).The maximum increase in sperm motility (but not in sperm concentration!) was observed in 24 men in 6 months – 49.6 (39.6; 56.4)% (p < 0.05). Percent of morphologically normal spermatozoa in ejaculate slightly increased only in 12 men in that time period from 0–1% to 1–2%. The total testosterone level was 2.4 times higher than the baseline (31.6±7.2 vs 13.2±4.3 nmol/l, p < 0.05), the free testosterone level was 1.8 times higher (14.5±3.5 vs 7.9±3.0 pgl/ml, p < 0.05). Unlike the PRP group, in the group of men without PRP treatment, the sperm parameters did not changed compared with the baseline in 4 months after the starting hormonal and AO treatment. A significant increase of sperin concentration was observed only in 17 of 35 patients in 6 months. Sperm motility and percent of morphologically normal spermatozoa after the treatment did not differ from the baseline. Changes in the testosterone levels were similar to changes in PRP group. Limitations, reasons for caution Only young and middle-aged men were considered in the study. Large randomized controlled studies are required to confirm the PRP therapy efficacy and safety of f various fertility disorders. There are also no standardized protocols for PRP preparation. Wider implications of the findings: PRP therapy may have great potential for the treatment of male infertility and improving spermatogenesis. Optimization of methods of PRP preparation and dosage of testicular injections can enhance reproductive outcomes in assisted reproductive technologies. Trial registration number Not applicable


2012 ◽  
Vol 24 (1) ◽  
pp. 170 ◽  
Author(s):  
A. Kouba ◽  
E. Willis ◽  
C. Vance ◽  
S. Hasenstab ◽  
S. Reichling ◽  
...  

Species-specific differences in breeding strategies and physiology have limited the application of assisted reproductive technologies (ART) for critically endangered amphibians in captive assurance colonies. In 2006, the Memphis Zoo (MZ) initiated a program to develop ART for the critically endangered Mississippi gopher frog after natural breeding failed. Standard gamete collection and IVF developed by MZ for reproducing endangered toads such as the Wyoming or boreal toad were applied to the gopher frog with little success, especially hormonal therapy for sperm production. Using the leopard frog as a model species for Ranids, we tested the time and dose dependence of a luteinizing hormone releasing hormone analogue (LHRHa) and hCG on sperm quantity and quality. Initial findings from the leopard frog study were critical in designing the study on gopher frogs. Our objectives were to (1) compare 2 different hormones administered intraperitoneal (500 IU hCG vs 15 μg LHRHa) or their combination on spermiation in gopher frogs; (2) develop in vivo oocyte maturation and ovulation protocols using LHRHa (15 μg) and hCG (500 IU); and (3) transfer this technology to another institution as proof of principle. In gopher frogs, 100 and 83% of the males produced sperm in response to the LHRHa and the combination treatment, respectively, whereas only 16% responded to hCG alone. Sperm concentration peaked at 1 h post-administration for all treatments, with the LHRH/hCG cocktail treatment producing the highest concentration of sperm (mean = 4.6 × 106 ± 1.2 × 106 sperm mL–1, n = 6). No differences in motility were observed between treatments (P > 0.05). For females, a series of priming hormones of hCG and LHRHa were given several months before an ovulatory hormone regimen resulting in ovulation by 100% of the females (n = 6), whereas animals not primed failed to ovulate (n = 4). These 3 separate priming and IVF trials conducted between 2008 and 2010 resulted in each female laying ∼2000 eggs, with an average fertilization rate of 76% for inseminated eggs and hundreds of tadpoles produced. These IVF tadpoles represent the first captive reproduction of gopher frogs and highlight how ART can be applied to conservation and genetic management of threatened species. Subsequently, we tested our IVF protocols on gopher frogs at Omaha's Henry Doorly Zoo using fresh (collected on site) and chilled, shipped sperm from MZ. We collected 6169 eggs from 9 hormone-primed females with all animals ovulating. A portion of the total eggs ovulated were inseminated, resulting in 2401 fertilized eggs (38.9% of total eggs collected) across 18 different male–female pairings leading to viable tadpoles. In addition, sperm transferred overnight from the MZ produced 202/441 fertilized eggs (46%). The transfer of this technology and production of endangered amphibians using chilled, shipped sperm from live animals is a conservation milestone that can be applied to other captive breeding programs.


2020 ◽  
Vol 32 (2) ◽  
pp. 203
Author(s):  
N. Buzzell ◽  
S. Blash ◽  
K. Miner ◽  
M. Schofield ◽  
J. Pollock ◽  
...  

The objective of this study was to investigate a method of oviducal semen deposition as a strategy for producing offspring from poor-quality cryopreserved goat sperm. Invitro fertilisation (IVF) and AI are common assisted reproductive technologies used in small ruminants, but they have varied results in the goat. The use of poor-quality cryopreserved-thawed sperm (<50% live/dead ratio at post-thaw) can decrease the rate of success. These procedures were performed in the month of November in Central Massachusetts in the United States (42° N). Seven 10-year-old dairy goats (Saanen, Toggenburg, and Alpine breeds) were synchronised and superovulated using a progesterone implant on Day 0, a prostaglandin injection at Day 7, two daily injections of 36mg of FSH ~12h apart on Days 12-15, and progesterone implant removal on Day 14 followed by an injection of 50µg of gonadotrophin-releasing hormone. Sperm deposition was performed on Day 17 (72 h after implant removal). The animals were anaesthetised using a standardised protocol, intubated, and maintained using isoflurane, and sterile prep was performed before a midline laparotomy procedure. Straws from a single ejaculate from a transgenic founder that was cryopreserved using a commercial two-step glycerol-egg yolk-based extender were used. A straw from this collection was post-thawed 30 days after collection and, using a commercial live/dead stain, 67% live sperm was determined. The optimal type of sperm prep and sperm concentration is unknown and may be dependent on sperm quality. Therefore, different gradient preps using Vitrolife SpermGrad at three volumes (1.5 (used on two animals), 1.0, and 0.5mL) as well as two volumes of IVF Bioscience Bovine BO-SemenPrep (4.0mL (used on two animals) and 2.0mL) were used. All five pellets were diluted in 1.0mL of IVF Bioscience Bovine BO-IVF media. Sperm concentrations ranging from 75×106 to 27×106 spermmL−1 were deposited into one oviduct; then, a 10:1 dilution was performed and 7.5×106 to 2.7×10 spermmL−1 were deposited into the contralateral oviduct. The depositions were performed just proximal to the uterotubal junction in a volume of 0.1mL of diluent via a tuberculin syringe attached to a 20-gauge needle. Two days following the procedure, oviducts were flushed postmortem from three of the seven randomly selected goats. All three had fertilised embryos, and nineteen 8-cell embryos were retrieved. Three of these embryos were surgically transferred to the distal uterine horn of a suitable recipient. The recipient became pregnant and produced a single offspring. The remaining four of seven goats were killed 41 days post-surgery. Two of the four goats were pregnant, with one carrying one fetus and the other carrying five fetuses. Further studies are needed to optimise this method, but these initial results indicate that oviducal semen deposition directly into the oviduct proximal to the uterotubal junction may be a suitable alternative for producing offspring from suboptimal cryopreserved-thawed goat sperm.


2017 ◽  
Vol 29 (1) ◽  
pp. 190
Author(s):  
A. M. Raseona ◽  
O. A. Ajao ◽  
L. D. Nethengwe ◽  
L. R. Madzhie ◽  
T. L. Nedambale ◽  
...  

Preservation of semen is an important process to ensure that semen quality is sufficient for assisted reproductive technologies. The aim of this study was to evaluate the viability of bull semen collected by electro-ejaculation using commercial semen extender and 2 modified culture media stored at controlled RT (24°C) for 72 h. Two Nguni bulls were used for semen collection; after collection, the semen was evaluated macroscopically for volume, pH, and colour, and microscopically for sperm motility, viability, and morphology. Uncontaminated semen samples with progressive motility >70% and morphological defects <20% were pooled after collection before being aliquoted into 3 extenders, namely Triladyl, modified Ham’s F10, and TCM-199 culture media, at a dilution ratio of 1:4 and then stored at controlled RT (24°C). Sperm motility rate was analysed using the computer-aided sperm analyser after 0, 24, 48, and 72 h of storage. Sperm morphology and viability was performed after staining the sperm cells with spermac and nigrosine-eosin stain, respectively. The study was replicated 4 times and data were analysed using ANOVA. Triladyl had a higher sperm viability rate (41.3%) and total motility rate (96.3%) for 72 h (P < 0.01) compared with the 2 modified culture media, Ham’s F10 (26.5 and 86.8%) and TCM-199 (25.0 and 86.7%), respectively. However, Ham’s F10 had higher progressive motility rate (37.8%) as compared with the other extenders, TCM-199 (31.7%) and Triladyl (23.4). There was no significant difference (P > 0.05), in viability rate between Ham’s F10 (26.5%) and TCM-199 (25.0%). No significant difference (P > 0.05) in straight line velocity was observed for the three extenders. Furthermore, no significant difference was observed in total sperm abnormalities, except for reacted acrosomes and absent tails (P > 0.05), between the 2 Nguni bulls. Nguni semen can be preserved in Triladyl or modified Ham’s F10 and TCM-199 culture media stored at 24°C and stay viable for 72 h. Triladyl proved to be the best suitable extender of the 3 extenders, showing higher sperm viability and total motility rate as compared with Ham’s F10 and TCM-199 modified culture media.


2020 ◽  
Vol 32 (2) ◽  
pp. 137
Author(s):  
Y. Pirosanto ◽  
M. Valera ◽  
A. Molina ◽  
J. Dorado ◽  
S. Demyda-Peyrás

Inbreeding depression, a genetic condition produced by the mating of close-related individuals, has been associated with a reduction of fertility in several species. However, a loss in sperm quality was also associated with age. In horses, the few existing reports have described a tendency of both parameters to produce a negative effect on sperm quality. However, those reports were performed using a subjective evaluation of sperm motility. In the present study, a total of 692 ejaculates from 86 Pure Spanish stallions (PRE), aged between 3 and 22 years, were evaluated using a computer-assisted methodology to determine the effect of inbreeding in four semen parameters: free-gel volume (V), sperm concentration (C, by haemocytometer), and total (TM) and progressive (PM) sperm motility (by Spermvision sperm class analyser; Minitube). The inbreeding coefficient (F) was estimated using 300 000 PRE pedigree records approximately (minimum pedigree depth, eight equivalent complete generations; range, between 1 and 30.1%). Stallion, age, ejaculate, and season of semen collection were the variables included in the statistical model (general linear model), with ejaculate and season being the variables with a major effect (by variance components analysis). Our results showed that sperm concentration (r=−0.18; P&lt;0.0001) and volume (to a lesser extent) were reduced with advancing age, both showing a major decline after 15 years of age. To the contrary, sperm motility was not affected by age of the stallion. We also found a negative correlation between the inbreeding coefficient and ejaculate volume (r=−0.14; P&lt;0.001), with a marked decrease seen when F was between 7 and 20%. Also, a negative correlation was observed in PM (r=−0.08; P&lt;0.05), although to a lower extent. Conversely, C and TM were not affected by inbreeding depression (P&gt;0.05). In conclusion, our results demonstrated that high levels of inbreeding can compromise severely the sperm quality of the PRE stallion, which, subsequently, may have a negative influence on fertility. Ongoing studies using genomic data will help to detect genetic variants associated with stallion semen quality and how it is influenced by inbreeding in specific genomic regions.


2019 ◽  
Vol 7 ◽  
pp. 2050313X1983415
Author(s):  
José Arturo Mora Rodríguez ◽  
Leonardo M Porchia ◽  
Felipe Camargo ◽  
Esther López-Bayghen

Male patients suffering from oligoasthenoteratozoospermia typically failed to achieve pregnancy, even with assisted reproductive technologies. Growth hormone and insulin-like growth factor 1 have been shown to regulate sperm quality parameters; therefore, the insulin-like growth factor 1 supplement could improve sperm parameters. Here, we determine the effect insulin-like growth factor 1 has on sperm parameters in a patient suffering from oligoasthenoteratozoospermia. A 47-year-old male was administered once a day 1.5 IU of insulin-like growth factor 1 by intradermal injection for 2 months. Seminogram analysis was performed before and after. Treatment with insulin-like growth factor 1 resulted in a 15.5-fold improvement in sperm concentration (1.1 × 106 vs 18.3 × 106 per mL), 71.4% change in volume (0.7 vs 1.2 mL), increased progressive motility (2% vs 43%), and the total volume of sperm with progressive motility (0% vs 23.6%). Here, we show that administering a daily dose of insulin-like growth factor 1 can improve sperm quality parameters.


Sign in / Sign up

Export Citation Format

Share Document