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Jogayle Howard - One of the best experts on this subject based on the ideXlab platform.

  • Recovery of gene diversity using long‐term cryopreserved spermatozoa and artificial insemination in the endangered black‐footed Ferret
    Animal Conservation, 2015
    Co-Authors: Jogayle Howard, Paul E Marinari, Rachel M Santymire, C. Lynch, David E. Wildt
    Abstract:

    One of the most significant challenges in the recovery of threatened species is the ability to maintain genetic diversity, avoid inbreeding and sustain population health and reproduction. Assisted reproductive techniques, including artificial insemination (AI), have been touted for decades as approaches that could contribute to the demographic and genetic management of rare species. Here, we report the first successful integration of AI with frozen semen into a formal recovery program and the positive impact on genetic diversity for the critically endangered Black-Footed Ferret Mustela nigripes. Techniques developed in the taxonomically related domestic Ferret Mustela putorius furo and Siberian polecat Mustela eversmannii were applied over time to selected Black-Footed Ferrets, including semen banking from six of the last 18 survivors. After evaluation, processing and storage in liquid nitrogen (−196°C/−321°F), for as long as 20 years, sperm samples were thawed and transabdominally inseminated into the uterine horns of female conspecifics. Eight Black-Footed Ferret offspring were produced using thawed sperm samples (including after two decades of cryopreservation) with inseminates containing as few as 3.4 × 106 motile spermatozoa. The incorporation of these offspring and/or their descendants into the ex situ breeding program prevented heterozygosity loss in the population and actually enhanced gene diversity (GD) significantly by 0.2% and lowered measures of inbreeding by 5.8%. This study demonstrates the utility and genetic diversity benefits of applying AI with cryopreserved spermatozoa 20 generations removed from the contemporary population for a wild animal revival program.

  • Genotypic and phenotypic consequences of reintroduction history in the Black-Footed Ferret (Mustela nigripes)
    Conservation Genetics, 2008
    Co-Authors: Samantha M. Wisely, Travis M Livieri, Sara Ann Mueting, Rachel M Santymire, Jogayle Howard
    Abstract:

    Population augmentation with translocated individuals has been shown to alleviate the effects of bottlenecks and drift. The first step to determine whether restoration for genetic considerations is warranted is to genetically monitor reintroduced populations and compare results to those from the source. To assess the need for genetic restoration, we evaluated genetic diversity and structure of reintroduced ( n  = 3) and captive populations of the endangered Black-Footed Ferret ( Mustela nigripes ). We measured genotypic changes among populations using seven microsatellite markers and compared phenotypic changes with eight morphometric characters. Results indicated that for the population which rapidly grew post-reintroduction, genetic diversity was equivalent to the captive, source population. When growth languished, only the population that was augmented yearly maintained diversity. Without augmentation, allelic diversity declined precipitously and phenotypic changes were apparent. Ferrets from the genetically depaupertate population had smaller limbs and smaller overall body size than Ferrets from the two populations with greater diversity. Population divergence ( F _ST = 0.10 ± 0.01) was surprisingly high given the common source of populations. Thus, it appears that 5–10 years of isolation resulted in both genotypic divergence and phenotypic changes to populations. We recommend translocation of 30–40 captive individuals per annum to reintroduction sites which have not become established quickly. This approach will maximize the retention of genetic diversity, yet maintain the beneficial effects of local adaptation without being swamped by immigration.

  • An unidentified filarial species and its impact on fitness in wild populations of the Black-Footed Ferret (Mustela nigripes).
    Journal of wildlife diseases, 2008
    Co-Authors: Samantha M. Wisely, Jogayle Howard, Rachel M Santymire, Steven A. Williams, Odile Bain, Katherine D. Bardsley, Elizabeth S Williams
    Abstract:

    Disease can threaten the restoration of endangered species directly by substantially decreasing host survival or indirectly via incremental decreases in survival and reproduction. During a biomedical survey of reintroduced populations of the highly endangered Black-Footed Ferret from 2002 to 2005, microfilariae discovered in the blood were putatively identified as Dirofilaria immitis, and widespread screening was initiated using a commercially available antigen-based ELISA test. A subset of animals (n = 16) was screened for D. immitis using a highly sensitive PCR-based assay. Microfilariae were also molecularly and morphologically characterized. Of 198 animals at six reintroduction sites, 12% had positive results using the ELISA test. No antigen-positive animals which were screened via PCR (n = 11) had positive PCR results, and all antigen-positive animals (n = 24) were asymptomatic. No significant differences were found in body mass of antigen-positive (male: 1223 +/- 82 g [mean +/- SD], female: 726 +/- 75 g) vs. antigen-negative (male: 1,198 +/- 119 g, female: 710 +/- 53 g) individuals (P = 0.4). Antigen prevalence was lower in juveniles (3%) than adults (12%; P = 0.03), and higher in in situ, captive-reared individuals (33%) than wild-born individuals (10%; P = 0.005). Morphologic analysis of microfilariae revealed they were neither D. immitis nor any other previously characterized North American species. PCR amplification of the 5S spacer region of rDNA revealed that the filarial sequence shared only 76% identity with D. immitis. This previously unidentified filarial sequence was present in all antigen positive animals (11 of 11 tested). It appears that Black-Footed Ferrets were infected with a previously undescribed species of filaria whose antigen cross-reacted with the ELISA assay, although further analysis is needed to make a conclusive statement. Nonetheless, this previously undescribed filaria does not appear to threaten recovery for this highly endangered mammal.

  • slow cooling prevents cold induced damage to sperm motility and acrosomal integrity in the black footed Ferret mustela nigripes
    Reproduction Fertility and Development, 2007
    Co-Authors: Paul E Marinari, J S Kreeger, Rachel M Santymire, David E. Wildt, Jogayle Howard
    Abstract:

    The endangered Black-Footed Ferret (Mustela nigripes) has benefited from artificial insemination; however, improved sperm cryopreservation protocols are still needed. The present study focused on identifying factors influencing gamete survival during processing before cryopreservation, including: (1) the presence or absence of seminal plasma; (2) temperature (25°C v. 37°C); (3) type of medium (Ham’s F10 medium v. TEST yolk buffer [TYB]); (4) cooling rate (slow, rapid and ultra-rapid); and (5) the presence or absence of glycerol. Seminal plasma did not compromise (P > 0.05) sperm motility or acrosomal integrity. Sperm motility traits were maintained longer (P 0.05) acrosomal integrity. Overall, TYB maintained optimal (P 0.05), indicating that 4% glycerol was non-toxic to Ferret sperm; however, glycerol failed to ameliorate the detrimental effects of either rapid or ultra-rapid cooling. The results of the present study demonstrate that the damage observed to Black-Footed Ferret spermatozoa is derived largely from the rate of cooling.

  • Slow cooling prevents cold-induced damage to sperm motility and acrosomal integrity in the Black-Footed Ferret (Mustela nigripes).
    Reproduction fertility and development, 2007
    Co-Authors: Rachel M Santymire, Paul E Marinari, J S Kreeger, David E. Wildt, Jogayle Howard
    Abstract:

    The endangered Black-Footed Ferret (Mustela nigripes) has benefited from artificial insemination; however, improved sperm cryopreservation protocols are still needed. The present study focused on identifying factors influencing gamete survival during processing before cryopreservation, including: (1) the presence or absence of seminal plasma; (2) temperature (25 degrees C v. 37 degrees C); (3) type of medium (Ham's F10 medium v. TEST yolk buffer [TYB]); (4) cooling rate (slow, rapid and ultra-rapid); and (5) the presence or absence of glycerol. Seminal plasma did not compromise (P > 0.05) sperm motility or acrosomal integrity. Sperm motility traits were maintained longer (P < 0.05) at 25 degrees C than at 37 degrees C in Ham's or TYB, but temperature did not affect (P > 0.05) acrosomal integrity. Overall, TYB maintained optimal (P < 0.05) sperm motility compared with Ham's medium, but Ham's medium maintained more (P < 0.05) intact acrosomes than TYB. Slow cooling (0.2 degrees C min(-1)) was optimal (P < 0.05) compared to rapid cooling (1 degrees C min(-1)), and ultra-rapid cooling (9 degrees C min(-1)) was found to be highly detrimental (P < 0.05). Results obtained in TYB with 0% or 4% glycerol were comparable (P > 0.05), indicating that 4% glycerol was non-toxic to Ferret sperm; however, glycerol failed to ameliorate the detrimental effects of either rapid or ultra-rapid cooling. The results of the present study demonstrate that the damage observed to Black-Footed Ferret spermatozoa is derived largely from the rate of cooling.

Rachel M Santymire - One of the best experts on this subject based on the ideXlab platform.

  • influence of vitamin e and carcass feeding supplementation on fecal glucocorticoid and androgen metabolites in male black footed Ferrets mustela nigripes
    PLOS ONE, 2020
    Co-Authors: Rachel M Santymire, Paul E Marinari, J S Kreeger, Shana R. Lavin, Michelle L. Rafacz, Heather Branvoldfaber, Judy P Checastaldo
    Abstract:

    In recent years, the ex situ population of the endangered Black-Footed Ferret (Mustela nigripes; Ferret) has experienced a decline in normal sperm morphology (from 50% to 20%), which may be linked to inbreeding depression and/or a dietary change. We examined the effects of adding carcass and vitamin E to the diet on stress and reproductive biomarkers in male Ferrets (n = 42 males including 16 juveniles and 26 adults) housed at the U.S. Fish and Wildlife National Black-Footed Ferret Conservation Center (Carr, CO, USA). Fecal samples (3x/week) were collected from November and December (pre-breeding season, no diet change), February through May (breeding season, diet change) and June (post-breeding season, diet change) and analyzed for fecal glucocorticoid metabolites (FGM) via a corticosterone enzyme immunoassay (EIA). A subset of samples from adult males (n = 15) were analyzed for fecal androgen metabolites (FAM) via a testosterone EIA. We first used a linear mixed effects model to identify the important fixed effects among meat treatment, vitamin E treatment, age class (juvenile or adult), and all possible interactions on each hormone. We then examined the important factor's effects across seasons using the non-parametric Friedman test. We found that age did not influence (p = 0.33) FGMs; however there was a significant effect of meat treatment on FGM (p = 0.04) and an effect of vitamin E on FAMs (p 0.05) between during and post-breeding season periods. Males that were not fed carcass had higher (p 0.05). Males supplemented with vitamin E had higher (p<0.001) FAM than non-supplemented males during the breeding season only. For both groups, FAM was highest (p<0.05) during the breeding season. In conclusion, adding carcass to the diet can reduce glucocorticoid production and adding vitamin E can increase testosterone during the breeding season, which may influence reproductive success.

  • Influence of vitamin E and carcass feeding supplementation on fecal glucocorticoid and androgen metabolites in male Black-Footed Ferrets (Mustela nigripes).
    PloS one, 2020
    Co-Authors: Rachel M Santymire, J S Kreeger, Heather Branvold-faber, Shana R. Lavin, Judy P. Che-castaldo, Michelle L. Rafacz, Paul E Marinari
    Abstract:

    In recent years, the ex situ population of the endangered Black-Footed Ferret (Mustela nigripes; Ferret) has experienced a decline in normal sperm morphology (from 50% to 20%), which may be linked to inbreeding depression and/or a dietary change. We examined the effects of adding carcass and vitamin E to the diet on stress and reproductive biomarkers in male Ferrets (n = 42 males including 16 juveniles and 26 adults) housed at the U.S. Fish and Wildlife National Black-Footed Ferret Conservation Center (Carr, CO, USA). Fecal samples (3x/week) were collected from November and December (pre-breeding season, no diet change), February through May (breeding season, diet change) and June (post-breeding season, diet change) and analyzed for fecal glucocorticoid metabolites (FGM) via a corticosterone enzyme immunoassay (EIA). A subset of samples from adult males (n = 15) were analyzed for fecal androgen metabolites (FAM) via a testosterone EIA. We first used a linear mixed effects model to identify the important fixed effects among meat treatment, vitamin E treatment, age class (juvenile or adult), and all possible interactions on each hormone. We then examined the important factor's effects across seasons using the non-parametric Friedman test. We found that age did not influence (p = 0.33) FGMs; however there was a significant effect of meat treatment on FGM (p = 0.04) and an effect of vitamin E on FAMs (p 0.05) between during and post-breeding season periods. Males that were not fed carcass had higher (p 0.05). Males supplemented with vitamin E had higher (p

  • implementing the use of a biobank in the endangered black footed Ferret mustela nigripes
    Reproduction Fertility and Development, 2016
    Co-Authors: Rachel M Santymire
    Abstract:

    In the current global health climate, many conservation biologists are managing crisis situations, including increased species extinction rates. One strategy for securing wildlife populations into the future is to preserve biomaterials in genome resource banks (GRB; or ‘biobanks’). However, for GRBs to be successful we must understand the fundamental reproductive biology of species, along with developing assisted reproductive techniques (ARTs), including AI and semen cryopreservation. ART has been successfully used for several taxa, from amphibians to mammals, including ungulates, carnivores and primates. Not all these success stories implemented the use of a biobank, but one example that discussed herein is the Black-Footed Ferret (Mustela nigripes) GRB. From a founder population of seven individuals, this species has been breeding in a managed setting for nearly 30 years. The goal of the breeding program is to maintain genetic integrity by ensuring each individual has the opportunity to pass his/her genes onto the next generation, while simultaneously providing animals for release into the wild. Scientists have used ART (e.g. AI) in the recovery program. Recently, semen from an individual of the founder population that was cryopreserved for up to 20 years was used successfully for AI, which improved the genetic diversity of the population. The Black-Footed Ferret recovery program can serve as a model for other endangered species and demonstrates the usefulness of ART and GRBs to maintain highly endangered species into the future.

  • A Road Map for 21st Century Genetic Restoration: Gene Pool Enrichment of the Black-Footed Ferret
    The Journal of heredity, 2015
    Co-Authors: Samantha M. Wisely, Rachel M Santymire, Oliver A. Ryder, John F. Engelhardt, Ben J. Novak
    Abstract:

    Interspecies somatic cell nuclear transfer (iSCNT) could benefit recovery programs of critically endangered species but must be weighed with the risks of failure. To weigh the risks and benefits, a decision-making process that evaluates progress is needed. Experiments that evaluate the efficiency and efficacy of blastocyst, fetal, and post-parturition development are necessary to determine the success or failure or species-specific iSCNT programs. Here, we use the Black-Footed Ferret (Mustela nigripes) as a case study for evaluating this emerging biomedical technology as a tool for genetic restoration. The Black-Footed Ferret has depleted genetic variation yet genome resource banks contain genetic material of individuals not currently represented in the extant lineage. Thus, genetic restoration of the species is in theory possible and could help reduce the persistent erosion of genetic diversity from drift. Extensive genetic, genomic, and reproductive science tools have previously been developed in Black-Footed Ferrets and would aid in the process of developing an iSCNT protocol for this species. Nonetheless, developing reproductive cloning will require years of experiments and a coordinated effort among recovery partners. The information gained from a well-planned research effort with the goal of genetic restoration via reproductive cloning could establish a 21st century model for evaluating and implementing conservation breeding that would be applicable to other genetically impoverished species.

  • Recovery of gene diversity using long‐term cryopreserved spermatozoa and artificial insemination in the endangered black‐footed Ferret
    Animal Conservation, 2015
    Co-Authors: Jogayle Howard, Paul E Marinari, Rachel M Santymire, C. Lynch, David E. Wildt
    Abstract:

    One of the most significant challenges in the recovery of threatened species is the ability to maintain genetic diversity, avoid inbreeding and sustain population health and reproduction. Assisted reproductive techniques, including artificial insemination (AI), have been touted for decades as approaches that could contribute to the demographic and genetic management of rare species. Here, we report the first successful integration of AI with frozen semen into a formal recovery program and the positive impact on genetic diversity for the critically endangered Black-Footed Ferret Mustela nigripes. Techniques developed in the taxonomically related domestic Ferret Mustela putorius furo and Siberian polecat Mustela eversmannii were applied over time to selected Black-Footed Ferrets, including semen banking from six of the last 18 survivors. After evaluation, processing and storage in liquid nitrogen (−196°C/−321°F), for as long as 20 years, sperm samples were thawed and transabdominally inseminated into the uterine horns of female conspecifics. Eight Black-Footed Ferret offspring were produced using thawed sperm samples (including after two decades of cryopreservation) with inseminates containing as few as 3.4 × 106 motile spermatozoa. The incorporation of these offspring and/or their descendants into the ex situ breeding program prevented heterozygosity loss in the population and actually enhanced gene diversity (GD) significantly by 0.2% and lowered measures of inbreeding by 5.8%. This study demonstrates the utility and genetic diversity benefits of applying AI with cryopreserved spermatozoa 20 generations removed from the contemporary population for a wild animal revival program.

Paul E Marinari - One of the best experts on this subject based on the ideXlab platform.

  • influence of vitamin e and carcass feeding supplementation on fecal glucocorticoid and androgen metabolites in male black footed Ferrets mustela nigripes
    PLOS ONE, 2020
    Co-Authors: Rachel M Santymire, Paul E Marinari, J S Kreeger, Shana R. Lavin, Michelle L. Rafacz, Heather Branvoldfaber, Judy P Checastaldo
    Abstract:

    In recent years, the ex situ population of the endangered Black-Footed Ferret (Mustela nigripes; Ferret) has experienced a decline in normal sperm morphology (from 50% to 20%), which may be linked to inbreeding depression and/or a dietary change. We examined the effects of adding carcass and vitamin E to the diet on stress and reproductive biomarkers in male Ferrets (n = 42 males including 16 juveniles and 26 adults) housed at the U.S. Fish and Wildlife National Black-Footed Ferret Conservation Center (Carr, CO, USA). Fecal samples (3x/week) were collected from November and December (pre-breeding season, no diet change), February through May (breeding season, diet change) and June (post-breeding season, diet change) and analyzed for fecal glucocorticoid metabolites (FGM) via a corticosterone enzyme immunoassay (EIA). A subset of samples from adult males (n = 15) were analyzed for fecal androgen metabolites (FAM) via a testosterone EIA. We first used a linear mixed effects model to identify the important fixed effects among meat treatment, vitamin E treatment, age class (juvenile or adult), and all possible interactions on each hormone. We then examined the important factor's effects across seasons using the non-parametric Friedman test. We found that age did not influence (p = 0.33) FGMs; however there was a significant effect of meat treatment on FGM (p = 0.04) and an effect of vitamin E on FAMs (p 0.05) between during and post-breeding season periods. Males that were not fed carcass had higher (p 0.05). Males supplemented with vitamin E had higher (p<0.001) FAM than non-supplemented males during the breeding season only. For both groups, FAM was highest (p<0.05) during the breeding season. In conclusion, adding carcass to the diet can reduce glucocorticoid production and adding vitamin E can increase testosterone during the breeding season, which may influence reproductive success.

  • Influence of vitamin E and carcass feeding supplementation on fecal glucocorticoid and androgen metabolites in male Black-Footed Ferrets (Mustela nigripes).
    PloS one, 2020
    Co-Authors: Rachel M Santymire, J S Kreeger, Heather Branvold-faber, Shana R. Lavin, Judy P. Che-castaldo, Michelle L. Rafacz, Paul E Marinari
    Abstract:

    In recent years, the ex situ population of the endangered Black-Footed Ferret (Mustela nigripes; Ferret) has experienced a decline in normal sperm morphology (from 50% to 20%), which may be linked to inbreeding depression and/or a dietary change. We examined the effects of adding carcass and vitamin E to the diet on stress and reproductive biomarkers in male Ferrets (n = 42 males including 16 juveniles and 26 adults) housed at the U.S. Fish and Wildlife National Black-Footed Ferret Conservation Center (Carr, CO, USA). Fecal samples (3x/week) were collected from November and December (pre-breeding season, no diet change), February through May (breeding season, diet change) and June (post-breeding season, diet change) and analyzed for fecal glucocorticoid metabolites (FGM) via a corticosterone enzyme immunoassay (EIA). A subset of samples from adult males (n = 15) were analyzed for fecal androgen metabolites (FAM) via a testosterone EIA. We first used a linear mixed effects model to identify the important fixed effects among meat treatment, vitamin E treatment, age class (juvenile or adult), and all possible interactions on each hormone. We then examined the important factor's effects across seasons using the non-parametric Friedman test. We found that age did not influence (p = 0.33) FGMs; however there was a significant effect of meat treatment on FGM (p = 0.04) and an effect of vitamin E on FAMs (p 0.05) between during and post-breeding season periods. Males that were not fed carcass had higher (p 0.05). Males supplemented with vitamin E had higher (p

  • Recovery of gene diversity using long‐term cryopreserved spermatozoa and artificial insemination in the endangered black‐footed Ferret
    Animal Conservation, 2015
    Co-Authors: Jogayle Howard, Paul E Marinari, Rachel M Santymire, C. Lynch, David E. Wildt
    Abstract:

    One of the most significant challenges in the recovery of threatened species is the ability to maintain genetic diversity, avoid inbreeding and sustain population health and reproduction. Assisted reproductive techniques, including artificial insemination (AI), have been touted for decades as approaches that could contribute to the demographic and genetic management of rare species. Here, we report the first successful integration of AI with frozen semen into a formal recovery program and the positive impact on genetic diversity for the critically endangered Black-Footed Ferret Mustela nigripes. Techniques developed in the taxonomically related domestic Ferret Mustela putorius furo and Siberian polecat Mustela eversmannii were applied over time to selected Black-Footed Ferrets, including semen banking from six of the last 18 survivors. After evaluation, processing and storage in liquid nitrogen (−196°C/−321°F), for as long as 20 years, sperm samples were thawed and transabdominally inseminated into the uterine horns of female conspecifics. Eight Black-Footed Ferret offspring were produced using thawed sperm samples (including after two decades of cryopreservation) with inseminates containing as few as 3.4 × 106 motile spermatozoa. The incorporation of these offspring and/or their descendants into the ex situ breeding program prevented heterozygosity loss in the population and actually enhanced gene diversity (GD) significantly by 0.2% and lowered measures of inbreeding by 5.8%. This study demonstrates the utility and genetic diversity benefits of applying AI with cryopreserved spermatozoa 20 generations removed from the contemporary population for a wild animal revival program.

  • The Black-Footed Ferret: on the brink of recovery?
    Advances in experimental medicine and biology, 2014
    Co-Authors: Rachel M Santymire, Travis M Livieri, Heather Branvold-faber, Paul E Marinari
    Abstract:

    In an attempt to save the species from extinction, the last remaining 18 Black-Footed Ferrets (Mustela nigripes) were trapped up from the wild to initiate a captive breeding program. Nearly 30 years later more than 8,000 Black-Footed Ferrets have been produced in captivity and approximately 4,100 animals have been reintroduced into 20 sites in eight US states (Arizona, New Mexico, Utah, Colorado, Kansas, Wyoming, South Dakota and Montana), Mexico and Canada. However, full recovery of the species has yet to be achieved, mainly due to limited viable habitat, disease and reduced fecundity. This chapter will highlight the advances in the Black-Footed Ferret recovery program over the last 10 years including: (1) adaptive management techniques employed for the captive population; (2) development of new reintroduction sites and associated challenges facing wild Black-Footed Ferrets; and (3) optimization of assisted reproductive techniques to secure the future of this rare species.

  • vaccination with f1 v fusion protein protects black footed Ferrets mustela nigripes against plague upon oral challenge with yersinia pestis
    Journal of Wildlife Diseases, 2008
    Co-Authors: Tonie E. Rocke, Paul E Marinari, J S Kreeger, Jeffrey T Enama, Susan R. Smith, Bradford S Powell
    Abstract:

    Previous studies have established that vaccination of Black-Footed Ferrets (Mustela nigripes) with F1-V fusion protein by subcutaneous (SC) injection protects the animals against plague upon injection of the bacterium Yersinia pestis. This study demonstrates that the F1-V antigen can also protect Ferrets against plague contracted via ingestion of a Y. pestis-infected mouse, a probable route for natural infection. Eight Black-Footed Ferret kits were vaccinated with F1-V protein by SC injection at approximately 60 days-of-age. A booster vaccination was administered 3 mo later via SC injection. Four additional Ferret kits received placebos. The animals were challenged 6 wk after the boost by feeding each one a Y. pestis-infected mouse. All eight vaccinates survived challenge, while the four controls succumbed to plague within 3 days after exposure. To determine the duration of antibody postvaccination, 18 additional Black-Footed Ferret kits were vaccinated and boosted with F1-V by SC injection at 60 and 120 days-of-age. High titers to both F1 and V (mean reciprocal titers of 18,552 and 99,862, respectively) were found in all vaccinates up to 2 yr postvaccination, whereas seven control animals remained antibody negative throughout the same time period.

David E. Wildt - One of the best experts on this subject based on the ideXlab platform.

  • Recovery of gene diversity using long‐term cryopreserved spermatozoa and artificial insemination in the endangered black‐footed Ferret
    Animal Conservation, 2015
    Co-Authors: Jogayle Howard, Paul E Marinari, Rachel M Santymire, C. Lynch, David E. Wildt
    Abstract:

    One of the most significant challenges in the recovery of threatened species is the ability to maintain genetic diversity, avoid inbreeding and sustain population health and reproduction. Assisted reproductive techniques, including artificial insemination (AI), have been touted for decades as approaches that could contribute to the demographic and genetic management of rare species. Here, we report the first successful integration of AI with frozen semen into a formal recovery program and the positive impact on genetic diversity for the critically endangered Black-Footed Ferret Mustela nigripes. Techniques developed in the taxonomically related domestic Ferret Mustela putorius furo and Siberian polecat Mustela eversmannii were applied over time to selected Black-Footed Ferrets, including semen banking from six of the last 18 survivors. After evaluation, processing and storage in liquid nitrogen (−196°C/−321°F), for as long as 20 years, sperm samples were thawed and transabdominally inseminated into the uterine horns of female conspecifics. Eight Black-Footed Ferret offspring were produced using thawed sperm samples (including after two decades of cryopreservation) with inseminates containing as few as 3.4 × 106 motile spermatozoa. The incorporation of these offspring and/or their descendants into the ex situ breeding program prevented heterozygosity loss in the population and actually enhanced gene diversity (GD) significantly by 0.2% and lowered measures of inbreeding by 5.8%. This study demonstrates the utility and genetic diversity benefits of applying AI with cryopreserved spermatozoa 20 generations removed from the contemporary population for a wild animal revival program.

  • slow cooling prevents cold induced damage to sperm motility and acrosomal integrity in the black footed Ferret mustela nigripes
    Reproduction Fertility and Development, 2007
    Co-Authors: Paul E Marinari, J S Kreeger, Rachel M Santymire, David E. Wildt, Jogayle Howard
    Abstract:

    The endangered Black-Footed Ferret (Mustela nigripes) has benefited from artificial insemination; however, improved sperm cryopreservation protocols are still needed. The present study focused on identifying factors influencing gamete survival during processing before cryopreservation, including: (1) the presence or absence of seminal plasma; (2) temperature (25°C v. 37°C); (3) type of medium (Ham’s F10 medium v. TEST yolk buffer [TYB]); (4) cooling rate (slow, rapid and ultra-rapid); and (5) the presence or absence of glycerol. Seminal plasma did not compromise (P > 0.05) sperm motility or acrosomal integrity. Sperm motility traits were maintained longer (P 0.05) acrosomal integrity. Overall, TYB maintained optimal (P 0.05), indicating that 4% glycerol was non-toxic to Ferret sperm; however, glycerol failed to ameliorate the detrimental effects of either rapid or ultra-rapid cooling. The results of the present study demonstrate that the damage observed to Black-Footed Ferret spermatozoa is derived largely from the rate of cooling.

  • Slow cooling prevents cold-induced damage to sperm motility and acrosomal integrity in the Black-Footed Ferret (Mustela nigripes).
    Reproduction fertility and development, 2007
    Co-Authors: Rachel M Santymire, Paul E Marinari, J S Kreeger, David E. Wildt, Jogayle Howard
    Abstract:

    The endangered Black-Footed Ferret (Mustela nigripes) has benefited from artificial insemination; however, improved sperm cryopreservation protocols are still needed. The present study focused on identifying factors influencing gamete survival during processing before cryopreservation, including: (1) the presence or absence of seminal plasma; (2) temperature (25 degrees C v. 37 degrees C); (3) type of medium (Ham's F10 medium v. TEST yolk buffer [TYB]); (4) cooling rate (slow, rapid and ultra-rapid); and (5) the presence or absence of glycerol. Seminal plasma did not compromise (P > 0.05) sperm motility or acrosomal integrity. Sperm motility traits were maintained longer (P < 0.05) at 25 degrees C than at 37 degrees C in Ham's or TYB, but temperature did not affect (P > 0.05) acrosomal integrity. Overall, TYB maintained optimal (P < 0.05) sperm motility compared with Ham's medium, but Ham's medium maintained more (P < 0.05) intact acrosomes than TYB. Slow cooling (0.2 degrees C min(-1)) was optimal (P < 0.05) compared to rapid cooling (1 degrees C min(-1)), and ultra-rapid cooling (9 degrees C min(-1)) was found to be highly detrimental (P < 0.05). Results obtained in TYB with 0% or 4% glycerol were comparable (P > 0.05), indicating that 4% glycerol was non-toxic to Ferret sperm; however, glycerol failed to ameliorate the detrimental effects of either rapid or ultra-rapid cooling. The results of the present study demonstrate that the damage observed to Black-Footed Ferret spermatozoa is derived largely from the rate of cooling.

  • Sperm viability in the Black-Footed Ferret (Mustela nigripes) is influenced by seminal and medium osmolality.
    Cryobiology, 2006
    Co-Authors: Rachel M Santymire, Paul E Marinari, J S Kreeger, David E. Wildt, Jogayle Howard
    Abstract:

    Fundamental knowledge of spermatozoa cryobiology can assist with optimizing cryopreservation protocols needed for genetic management of the endangered Black-Footed Ferret. Objectives were to characterize semen osmolality and assess the influence of two media at various osmolalities on sperm viability. We examined the influence of Ham's F10 +Hepes medium (H) at 270, 400, 500 or 700 mOsm (adjusted with sucrose, a nonpermeating cryoprotectant) and TEST Yolk Buffer (TYB) with 0% (300 mOsm) versus 4% (900 mOsm) glycerol (a permeating cryoprotectant). Electroejaculates (n=16) were assessed for osmolality using a vapor pressure osmometer. For media comparison, semen (n=5) was collected in TYB 0%, split into six aliquots, and diluted in H270, H400, H500, H700, and TYB 0% or TYB 4%. Each sample was centrifuged (300 g, 8 min), resuspended in respective medium, and maintained at 37 degrees C for 3h. Sperm motility and forward progression were monitored every 30 min for 3h post-washing. Acrosomal integrity was monitored at 0 and 60 min post-washing. Results demonstrated that Black-Footed Ferret semen has a comparatively high osmolality (mean+/-SEM, 513.1+/-32.6 mOsm; range, 366-791 mOsm). Ferret spermatozoa were sensitive to hyperosmotic stress. Specifically, sperm motility was more susceptible (P0.05) by hypotonic solutions. Exposure to TYB 4% glycerol retained more (P

  • age dependent changes in sperm production semen quality and testicular volume in the black footed Ferret mustela nigripes
    Biology of Reproduction, 2000
    Co-Authors: Paul E Marinari, J S Kreeger, K.n. Wolf, Astrid Vargas, Mary Ann Ottinger, David E. Wildt, Jogayle Howard
    Abstract:

    The Black-Footed Ferret (Mustela nigripes), which was extirpated from its native North American prairie habitat during the 1980s, is being reintroduced to the wild because of a successful captive-breeding program. To enhance propagation, the reproductive biology of this endangered species is being studied intensively. The typical life span of the Black-Footed Ferret is approximately 7 yr. Female fecundity declines after 3 yr of age, but the influence of age on male reproduction is unknown. In this study, testis volume, seminal traits, sperm morphology, and serum testosterone were compared in 116 males from 1 to 7 yr of age living in captivity. Results demonstrated that testes volume during the peak breeding season was similar (P > 0.05) among males 1 to 5 yr of age, reduced (P 0.05) in older males. Histological comparison of testicular/epididymal tissue from 5- and 7-yr-old Black-Footed Ferrets confirmed that the interval between these two ages may represent a transitional period to reproductive senescence. In summary, functional reproductive capacity of male Black-Footed Ferrets exceeds that of females by at least 2 yr. Testes and seminal quality are indistinguishable among males 1 to 5 yr of age, with progressive reproductive aging occurring thereafter.

J S Kreeger - One of the best experts on this subject based on the ideXlab platform.

  • Influence of vitamin E and carcass feeding supplementation on fecal glucocorticoid and androgen metabolites in male Black-Footed Ferrets (Mustela nigripes).
    PloS one, 2020
    Co-Authors: Rachel M Santymire, J S Kreeger, Heather Branvold-faber, Shana R. Lavin, Judy P. Che-castaldo, Michelle L. Rafacz, Paul E Marinari
    Abstract:

    In recent years, the ex situ population of the endangered Black-Footed Ferret (Mustela nigripes; Ferret) has experienced a decline in normal sperm morphology (from 50% to 20%), which may be linked to inbreeding depression and/or a dietary change. We examined the effects of adding carcass and vitamin E to the diet on stress and reproductive biomarkers in male Ferrets (n = 42 males including 16 juveniles and 26 adults) housed at the U.S. Fish and Wildlife National Black-Footed Ferret Conservation Center (Carr, CO, USA). Fecal samples (3x/week) were collected from November and December (pre-breeding season, no diet change), February through May (breeding season, diet change) and June (post-breeding season, diet change) and analyzed for fecal glucocorticoid metabolites (FGM) via a corticosterone enzyme immunoassay (EIA). A subset of samples from adult males (n = 15) were analyzed for fecal androgen metabolites (FAM) via a testosterone EIA. We first used a linear mixed effects model to identify the important fixed effects among meat treatment, vitamin E treatment, age class (juvenile or adult), and all possible interactions on each hormone. We then examined the important factor's effects across seasons using the non-parametric Friedman test. We found that age did not influence (p = 0.33) FGMs; however there was a significant effect of meat treatment on FGM (p = 0.04) and an effect of vitamin E on FAMs (p 0.05) between during and post-breeding season periods. Males that were not fed carcass had higher (p 0.05). Males supplemented with vitamin E had higher (p

  • influence of vitamin e and carcass feeding supplementation on fecal glucocorticoid and androgen metabolites in male black footed Ferrets mustela nigripes
    PLOS ONE, 2020
    Co-Authors: Rachel M Santymire, Paul E Marinari, J S Kreeger, Shana R. Lavin, Michelle L. Rafacz, Heather Branvoldfaber, Judy P Checastaldo
    Abstract:

    In recent years, the ex situ population of the endangered Black-Footed Ferret (Mustela nigripes; Ferret) has experienced a decline in normal sperm morphology (from 50% to 20%), which may be linked to inbreeding depression and/or a dietary change. We examined the effects of adding carcass and vitamin E to the diet on stress and reproductive biomarkers in male Ferrets (n = 42 males including 16 juveniles and 26 adults) housed at the U.S. Fish and Wildlife National Black-Footed Ferret Conservation Center (Carr, CO, USA). Fecal samples (3x/week) were collected from November and December (pre-breeding season, no diet change), February through May (breeding season, diet change) and June (post-breeding season, diet change) and analyzed for fecal glucocorticoid metabolites (FGM) via a corticosterone enzyme immunoassay (EIA). A subset of samples from adult males (n = 15) were analyzed for fecal androgen metabolites (FAM) via a testosterone EIA. We first used a linear mixed effects model to identify the important fixed effects among meat treatment, vitamin E treatment, age class (juvenile or adult), and all possible interactions on each hormone. We then examined the important factor's effects across seasons using the non-parametric Friedman test. We found that age did not influence (p = 0.33) FGMs; however there was a significant effect of meat treatment on FGM (p = 0.04) and an effect of vitamin E on FAMs (p 0.05) between during and post-breeding season periods. Males that were not fed carcass had higher (p 0.05). Males supplemented with vitamin E had higher (p<0.001) FAM than non-supplemented males during the breeding season only. For both groups, FAM was highest (p<0.05) during the breeding season. In conclusion, adding carcass to the diet can reduce glucocorticoid production and adding vitamin E can increase testosterone during the breeding season, which may influence reproductive success.

  • vaccination with f1 v fusion protein protects black footed Ferrets mustela nigripes against plague upon oral challenge with yersinia pestis
    Journal of Wildlife Diseases, 2008
    Co-Authors: Tonie E. Rocke, Paul E Marinari, J S Kreeger, Jeffrey T Enama, Susan R. Smith, Bradford S Powell
    Abstract:

    Previous studies have established that vaccination of Black-Footed Ferrets (Mustela nigripes) with F1-V fusion protein by subcutaneous (SC) injection protects the animals against plague upon injection of the bacterium Yersinia pestis. This study demonstrates that the F1-V antigen can also protect Ferrets against plague contracted via ingestion of a Y. pestis-infected mouse, a probable route for natural infection. Eight Black-Footed Ferret kits were vaccinated with F1-V protein by SC injection at approximately 60 days-of-age. A booster vaccination was administered 3 mo later via SC injection. Four additional Ferret kits received placebos. The animals were challenged 6 wk after the boost by feeding each one a Y. pestis-infected mouse. All eight vaccinates survived challenge, while the four controls succumbed to plague within 3 days after exposure. To determine the duration of antibody postvaccination, 18 additional Black-Footed Ferret kits were vaccinated and boosted with F1-V by SC injection at 60 and 120 days-of-age. High titers to both F1 and V (mean reciprocal titers of 18,552 and 99,862, respectively) were found in all vaccinates up to 2 yr postvaccination, whereas seven control animals remained antibody negative throughout the same time period.

  • slow cooling prevents cold induced damage to sperm motility and acrosomal integrity in the black footed Ferret mustela nigripes
    Reproduction Fertility and Development, 2007
    Co-Authors: Paul E Marinari, J S Kreeger, Rachel M Santymire, David E. Wildt, Jogayle Howard
    Abstract:

    The endangered Black-Footed Ferret (Mustela nigripes) has benefited from artificial insemination; however, improved sperm cryopreservation protocols are still needed. The present study focused on identifying factors influencing gamete survival during processing before cryopreservation, including: (1) the presence or absence of seminal plasma; (2) temperature (25°C v. 37°C); (3) type of medium (Ham’s F10 medium v. TEST yolk buffer [TYB]); (4) cooling rate (slow, rapid and ultra-rapid); and (5) the presence or absence of glycerol. Seminal plasma did not compromise (P > 0.05) sperm motility or acrosomal integrity. Sperm motility traits were maintained longer (P 0.05) acrosomal integrity. Overall, TYB maintained optimal (P 0.05), indicating that 4% glycerol was non-toxic to Ferret sperm; however, glycerol failed to ameliorate the detrimental effects of either rapid or ultra-rapid cooling. The results of the present study demonstrate that the damage observed to Black-Footed Ferret spermatozoa is derived largely from the rate of cooling.

  • Slow cooling prevents cold-induced damage to sperm motility and acrosomal integrity in the Black-Footed Ferret (Mustela nigripes).
    Reproduction fertility and development, 2007
    Co-Authors: Rachel M Santymire, Paul E Marinari, J S Kreeger, David E. Wildt, Jogayle Howard
    Abstract:

    The endangered Black-Footed Ferret (Mustela nigripes) has benefited from artificial insemination; however, improved sperm cryopreservation protocols are still needed. The present study focused on identifying factors influencing gamete survival during processing before cryopreservation, including: (1) the presence or absence of seminal plasma; (2) temperature (25 degrees C v. 37 degrees C); (3) type of medium (Ham's F10 medium v. TEST yolk buffer [TYB]); (4) cooling rate (slow, rapid and ultra-rapid); and (5) the presence or absence of glycerol. Seminal plasma did not compromise (P > 0.05) sperm motility or acrosomal integrity. Sperm motility traits were maintained longer (P < 0.05) at 25 degrees C than at 37 degrees C in Ham's or TYB, but temperature did not affect (P > 0.05) acrosomal integrity. Overall, TYB maintained optimal (P < 0.05) sperm motility compared with Ham's medium, but Ham's medium maintained more (P < 0.05) intact acrosomes than TYB. Slow cooling (0.2 degrees C min(-1)) was optimal (P < 0.05) compared to rapid cooling (1 degrees C min(-1)), and ultra-rapid cooling (9 degrees C min(-1)) was found to be highly detrimental (P < 0.05). Results obtained in TYB with 0% or 4% glycerol were comparable (P > 0.05), indicating that 4% glycerol was non-toxic to Ferret sperm; however, glycerol failed to ameliorate the detrimental effects of either rapid or ultra-rapid cooling. The results of the present study demonstrate that the damage observed to Black-Footed Ferret spermatozoa is derived largely from the rate of cooling.