scholarly journals The effect of injection and topical application of hCG and GnRH agonist to induce sperm-release in the roseate frog, Geocrinia rosea

2020 ◽  
Vol 8 (1) ◽  
Author(s):  
Aimee J Silla ◽  
J Dale Roberts ◽  
Phillip G Byrne
Keyword(s):  
Topical Application ◽  
Disease Transmission ◽  
Sperm Concentration ◽  
Reproductive Technologies ◽  
Amphibian Conservation ◽  
Genetic Management ◽  
Breeding Programs ◽  
Hormone Administration ◽  
Applied Dose ◽  
Conservation Breeding

Abstract Reproductive technologies may assist amphibian conservation breeding programs (CBPs) to achieve propagation targets and genetic management goals. However, a trial-and-error approach to protocol refinement has led to few amphibian CBPs routinely employing reproductive technologies with predictable outcomes. Additionally, while injections can be safely administered to amphibians, perceived animal welfare risks, such as injury and disease transmission, warrant the development of alternative hormone administration protocols. The present study investigated the spermiation response of roseate frogs, Geocrinia rosea, administered various doses of human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone agonist (GnRH-a) via subcutaneous injection. This study also quantified the spermiation response of frogs administered both hormones via topical application. Total sperm, sperm concentration and sperm viability were assessed over a 12-h period post hormone administration. Males released sperm in response to the injection of hCG (88–100% response; 5, 10 or 20 IU), but all samples collected from males administered hCG topically (100, 100 + DMSO or 200 IU hCG) were aspermic. In contrast, males consistently released sperm in response to both the injection (100% response; 1, 5 or 10 μg), or topical application (80–100% response; 50, 50 + DMSO or 100 μg) of GnRH-a. Overall, the administration of GnRH-a was more effective at inducing spermiation than hCG. Mean total sperm and sperm concentration were highest in response to the optimal topically applied dose of 100 μg GnRH-a (mean total sperm = 2.44 × 103, sperm concentration = 1.48 × 105 sperm/ml). We provide novel evidence that topical application provides a viable alternative to injection for the administration of GnRH-a to induce spermiation in amphibians.

The Role of Reproductive Technologies in Amphibian Conservation Breeding Programs

2019 ◽  
Vol 7 (1) ◽  
pp. 499-519 ◽  
Author(s):  
Aimee J. Silla ◽  
Phillip G. Byrne
Keyword(s):  
Reproductive Technologies ◽  
Ex Situ ◽  
Amphibian Conservation ◽  
Genetic Management ◽  
Species Extinction ◽  
Amphibian Species ◽  
Breeding Programs ◽  
Biodiversity Crisis ◽  
Conservation Breeding

Anthropogenic environmental change has led to unprecedented rates of species extinction, presenting a major threat to global biodiversity. Among vertebrates, amphibians have been most severely impacted, with an estimated 41% of species now threatened with extinction. In response to this biodiversity crisis, a moral and ethical obligation exists to implement proactive interventionist conservation actions to assist species recovery and decelerate declines. Conservation breeding programs have been successfully established for several threatened amphibian species globally, aiming to prevent species’ extinction by maintaining genetically representative assurance colonies ex situ while providing individuals for population augmentation, translocation, and reestablishment in situ. Reproductive technologies have enormous potential to enhance the propagation and genetic management of threatened species. In this review, we discuss the role of reproductive technologies in amphibian conservation breeding programs and summarize technological advancements in amphibian hormone therapies, gamete storage, and artificial fertilization.

116 DEVELOPMENT OF ASSISTED REPRODUCTION TECHNOLOGIES FOR THE ENDANGERED MISSISSIPPI GOPHER FROG (RANA SEVOSA) AND SPERM TRANSFER FOR IN VITRO FERTILIZATION

2012 ◽  
Vol 24 (1) ◽  
pp. 170 ◽  
Author(s):  
A. Kouba ◽  
E. Willis ◽  
C. Vance ◽  
S. Hasenstab ◽  
S. Reichling ◽  
...  
Keyword(s):  
Captive Breeding ◽  
Assisted Reproductive Technologies ◽  
Sperm Concentration ◽  
Reproductive Technologies ◽  
Critically Endangered ◽  
Leopard Frog ◽  
Genetic Management ◽  
Mississippi Gopher Frog

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.

scholarly journals The Current and Future Uses of Biotechnology in Animal Agriculture

Ceiba ◽  
2016 ◽  
Vol 54 (1) ◽  
pp. 72-81
Author(s):  
Alison L. Van Eenennaam
Keyword(s):  
Genetic Engineering ◽  
Genetic Improvement ◽  
Production Systems ◽  
Reproductive Technologies ◽  
Past Century ◽  
Breeding Programs ◽  
Animal Products ◽  
Genetic Modifications ◽  
Genomic Tools

Biotechnologies have been an integral part of improvements in animal genetics, nutrition and health over the past century. Many biotechnologies have become fundamental components of efficient livestock production systems. The genetic improvements that have been enabled by biotechnologies have dramatically decreased the environmental footprint of animal protein production in many parts of the world, and continued innovation is required to address the projected increase in demand for animal products in the future. Breeding programs increasingly utilize a combination of advanced reproductive technologies and genomic tools to accelerate the rate of genetic gain by manipulating components of the breeder’s equation. The use of these biotechnologies and breeding methods has met with little public opposition. In contrast, the use of modern biotechnologies, defined as those that employ the use of in vitro nucleic acid techniques, have been highly controversial, especially those involving the use of genetic engineering. This modern biotechnology distinction is somewhat arbitrary as there are a number of biotechnologies that involve the use of in vitro processes, and many result in genetic modifications that are indistinguishable from the naturally-occurring variation that is the driver of both traditional breeding programs and evolution. A number of useful traits including disease resistance and animal welfare traits have been successfully introduced into various livestock species using both genetic engineering and gene editing techniques. Ultimately these techniques complement the genetic improvement that can be accomplished using traditional selection techniques and, if judged acceptable, offer an opportunity to synergistically accelerate genetic improvement in food animal species.

P–050 The effectiveness of the platelet-rich plasma treatment of men with severe oligoasthenoteratozoospermia

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
O Somova ◽  
H Ivanova ◽  
N Sotnyk ◽  
K Kovalenko ◽  
I Feskova
Keyword(s):  
Male Infertility ◽  
Sperm Motility ◽  
Testosterone Level ◽  
Assisted Reproductive Technologies ◽  
Platelet Rich Plasma ◽  
Sperm Concentration ◽  
Free Testosterone ◽  
Reproductive Technologies ◽  
Positive Effects ◽  
Free Testosterone Level

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

scholarly journals Deciphering genetic mate choice: Not so simple in group‐housed conservation breeding programs

2020 ◽  
Vol 13 (9) ◽  
pp. 2179-2189
Author(s):  
Katherine A. Farquharson ◽  
Carolyn J. Hogg ◽  
Katherine Belov ◽  
Catherine E. Grueber
Keyword(s):  
Mate Choice ◽  
Breeding Programs ◽  
Conservation Breeding

246 IN VITRO PRODUCTION OF SPRINGBOK (ANTIDORCAS MARSUPIALIS) EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 239 ◽  
Author(s):  
R. Krisher ◽  
A. Auer ◽  
K. Clark ◽  
K. Emsweller ◽  
S. Rogers ◽  
...  
Keyword(s):  
South Africa ◽  
Oocyte Maturation ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Genetic Management ◽  
Blastocyst Development ◽  
Antidorcas Marsupialis

The objective of this experiment was to develop in vitro embryo production (IVP) technologies in springbok (Antidorcas marsupialis), a southern African antelope. Springbok, a fairly common species on game farms in parts of South Africa, may be used as a model species for gamete rescue and IVP techniques to be applied to the conservation of other threatened antelope species. Springbok belong to the family bovidae, subfamily antilopinae, tribe antilopini, which comprises about twenty species in genera Gazella, Antilope, Procapra, Antidorcas, Litocranius, and Ammodorcas. In this tribe alone, there are 4 species or subspecies that are critically endangered, 3 that are endangered, and 10 that are considered vulnerable, demonstrating the need for antelope conservation efforts. In addition, our studies contributed to the South African biological resource bank, so that banked springbok semen and embryos might be used in the future for managed genetic contribution to isolated captive or wild populations via assisted reproductive technologies. Oocytes were recovered (3 replicates) from ovaries obtained at supervised culls for management purposes in South Africa, and cultured in defined Gmat or undefined TCM-199 with FCS maturation medium for 28-30 h (Brad et al. 2004 Reprod. Fertil. Dev. 16, 223). Oocytes were fertilized with frozen-thawed springbok epididymal spermatozoa in modified SOF fertilization medium with caffeine (Herrick et al. 2004 Biol. Reprod. 71, 948–958). Eighteen hours after insemination, a randomly selected subset of the zygotes were fixed to determine fertilization success. The remaining zygotes were cultured in G1/G2 media. On Day 7 of culture, embryos were analyzed for development to the morula or blastocyst stage. A total of 259 selected oocytes were collected from 50 females (5.2 selected oocytes/female on average). There was no difference in the percentage of oocytes normally fertilized (2 pronuclei, PN) between oocytes matured in Gmat (n= 43; 12%) and those matured in TCM-199 (n= 42; 10%). There were significantly (P < 0.05) more oocytes penetrated (e2 PN) when matured in TCM (50%) compared to Gmat (23%). There were no differences in embryonic cleavage or morula/blastocyst development (of total oocytes inseminated) between treatments (Gmat,n= 89, 54%, 9.0%; TCM-199, n= 85, 68%, 9.4%, respectively). In both treatments, the average blastocyst grade was 2.125 using the standard bovine grading system (Curtis, Cattle Embryo Transfer Procedure, 1991). In conclusion, in vitro oocyte maturation, fertilization, and embryo culture to the blastocyst stage is possible in springbok. Importantly, blastocysts can be produced in vitro under semi-defined conditions, demonstrating that oocyte maturation without serum does support developmental competence. This is important for the potential international movement of IVP embryos to be used for genetic management in the conservation of antelope species.

154 A method of oviductal semen deposition for use in the goat

2020 ◽  
Vol 32 (2) ◽  
pp. 203
Author(s):  
N. Buzzell ◽  
S. Blash ◽  
K. Miner ◽  
M. Schofield ◽  
J. Pollock ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
The United States ◽  
Reproductive Technologies ◽  
Poor Quality ◽  
Implant Removal ◽  
Suitable Alternative ◽  
Midline Laparotomy ◽  
Post Surgery

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.

scholarly journals Sexual Dimorphism for Coping Styles Complements Traditional Methods for Sex Determination in a Multivariety Endangered Hen Breed

Animals ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 1165
Author(s):  
Carlos Iglesias Pastrana ◽  
Francisco Javier Navas González ◽  
Carmen Marín Navas ◽  
Ander Arando Arbulu ◽  
Antonio González Ariza ◽  
...  
Keyword(s):  
Sex Determination ◽  
Coping Styles ◽  
Explanatory Power ◽  
Sex Distribution ◽  
Genetic Management ◽  
Breeding Programs ◽  
Sex Assignment ◽  
Population Conservation ◽  
Hardy Weinberg Equilibrium ◽  
Combining Methods

Sex determination is key to designing endangered poultry population conservation and breeding programs when sex distribution departs from Hardy–Weinberg equilibrium. A total of 112 Utrerana chickens (28 per variety, partridge, black, white, and franciscan) were selected for hatching day sexing. Sex assignation was performed through 10 methods. Three sex assignment criteria comprised criteria found in literature, opposite criteria to that in the literature, and composite criteria combining methods reporting the highest predictive success from the previous ones. This study aims to determine which method combinations may more successfully determine sex across the four varieties of Utrerana endangered hen breed to tailor noninvasive early specific models to determine sex in local chicken populations. Although the explanatory power of the three assignation criteria is equal (75%), assignation criteria 2 resulted to be the most efficient as it correctly assigns males more frequently. Only methods 3 (English method), 5 (general down feathers coloration), 7 (wing fan), and 10 (behavior/coping styles) reported significant differences regardless of the variety, hence, are appropriate for early sexing. Sex confirmation was performed at 1.5 months old. Identifying sex proportions enhances genetic management tasks in endangered populations, complementing more standardized techniques, which may result inefficient given the implicit diversity found in local populations.

scholarly journals Genetic management strategies for controlling infectious diseases in livestock populations

2002 ◽  
Vol 2002 ◽  
pp. 55-55 ◽  
Author(s):  
S. C. Bishop ◽  
K. Mackenzie
Keyword(s):  
Disease Resistance ◽  
Resistance Genes ◽  
Disease Transmission ◽  
Management Strategies ◽  
Disease Resistance Genes ◽  
Breeding Strategies ◽  
Genetic Management ◽  
Control Disease ◽  
The Impact ◽  
Prp Gene

Disease resistance is often cited as the major challenge facing animal geneticists, with much effort directed towards finding disease-resistance genes. The PrP gene controlling resistance of sheep to scrapie is such an example. To design effective breeding strategies utilising such genes, it is critical to understand the impact that these genes have upon disease transmission. For example, it has been shown that it is not necessary to make all animals genetically resistant in order to protect the population as a whole from epidemics (MacKenzie and Bishop, 1999). Additionally, concern is often voiced over the possibility of the pathogen co-evolving with the host, reducing the utility of the genes. By combining animal breeding and epidemiology theory, this study derives strategies for using disease resistance genes to control disease transmission, and considers the co-evolution risks with such strategies.

scholarly journals The use of insulin-like growth factor 1 improved the parameters of the seminogram in a patient with severe oligoasthenoteratozoospermia

2019 ◽  
Vol 7 ◽  
pp. 2050313X1983415
Author(s):  
José Arturo Mora Rodríguez ◽  
Leonardo M Porchia ◽  
Felipe Camargo ◽  
Esther López-Bayghen
Keyword(s):  
Growth Factor ◽  
Assisted Reproductive Technologies ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Reproductive Technologies ◽  
Quality Parameters ◽  
Intradermal Injection ◽  
Sperm Parameters ◽  
Insulin Like Growth Factor ◽  
Progressive Motility

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.

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