Liquid storage of stallion spermatozoa – Past, present and future

Clulow, J.R.; Gibb, Zamira

doi:10.1016/j.anireprosci.2022.107088

Cited by 9 publications

(8 citation statements)

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“…Thus, a simple and accurate method of determining pregnancy probability from a single mating event is of great value to the industry. Proteomic analyses have revealed that horse spermatozoa utilise a different ATP production and energy avenue to humans (Gibb et al 2014;Griffin et al 2022) and such knowledge has led to the development of the Equility device which uses dismount semen samples to predict pregnancy outcome in the field (Clulow and Gibb 2022) and guided development of sperm storage media for use at ambient temperatures (Aitken et al 2014). The Equility device provides a fluorescent reading proportional to mitochondrial oxidative phosphorylation, making this technology pivotal in breeder decision-making to remate pairs prior to the end of the breeding season where necessary.…”

Section: Fundamental and Applied Reproductive Engineeringmentioning

confidence: 99%

Recent advances in reproductive research in Australia and New Zealand: highlights from the Annual Meeting of the Society for Reproductive Biology, 2022

Green,

Chan,

Frost

et al. 2024

Reprod. Fertil. Dev.

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In 2022, the Society for Reproductive Biology came together in Christchurch New Zealand (NZ), for its first face-to-face meeting since the global COVID-19 pandemic. The meeting showcased recent advancements in reproductive research across a diverse range of themes relevant to human health and fertility, exotic species conservation, and agricultural breeding practices. Here, we highlight the key advances presented across the main themes of the meeting, including advances in addressing opportunities and challenges in reproductive health related to First Nations people in Australia and NZ; increasing conservation success of exotic species, including ethical management of invasive species; improvements in our understanding of developmental biology, specifically seminal fluid signalling, ovarian development and effects of environmental impacts such as endocrine-disrupting chemicals; and leveraging scientific breakthroughs in reproductive engineering to drive solutions for fertility, including in assisted reproductive technologies in humans and agricultural industries, and for regenerative medicine.

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Section: Fundamental and Applied Reproductive Engineeringmentioning

confidence: 99%

Recent advances in reproductive research in Australia and New Zealand: highlights from the Annual Meeting of the Society for Reproductive Biology, 2022

Green,

Chan,

Frost

et al. 2024

Reprod. Fertil. Dev.

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“…The optimal temperature for storing stallion semen has not been fully determined, but various reports suggest it ranges from 5 C to 20 C. 11,18 For fresh semen, it is recommended that the cooling rate during the phase transition of the sperm membrane is À0.5 C/min, and faster cooling rates may induce cold shock, which is detrimental to sperm viability. The mean temperature during December in New Zealand ranges between 20 C and 25 C (niwa.co.nz), and overnight the temperature ranges from 10 C to 17 C, therefore providing an ambient temperature within the range suitable for sperm storage.…”

Section: Introductionmentioning

confidence: 99%

“…More recently, it has been reported that post thaw motility >30% can be maintained at 5°C for 24 h and pregnancies 9 (9/16 cycles, 56%) have been achieved after thawing, centrifuging and storing frozen–thawed stallion semen at 5°C for up to 24 h 9,10 . In addition, new methods for the liquid storage of fresh stallion spermatozoa at room temperature in protein‐free media 11 may provide opportunities for improving the longevity of frozen–thawed spermatozoa.…”

Section: Introductionmentioning

confidence: 99%

“…The optimal temperature for storing stallion semen has not been fully determined, but various reports suggest it ranges from 5°C to 20°C 11,18 . For fresh semen, it is recommended that the cooling rate during the phase transition of the sperm membrane is −0.5°C/min, and faster cooling rates may induce cold shock, which is detrimental to sperm viability.…”

Section: Introductionmentioning

confidence: 99%

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A simplified fixed‐time insemination protocol using frozen–thawed stallion spermatozoa stored at 17°C for up to 24 h before insemination

Morris,

Harteveld,

Gibb

2024

Equine Veterinary Journal

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BackgroundInsemination of mares with frozen–thawed spermatozoa requires intensive management and results in 40%–60% per cycle pregnancy rates.ObjectivesTo determine if satisfactory fertility is possible for frozen–thawed semen after processing it through a microfluidic device, followed by storage at 17°C for up to 24 h before fixed‐time insemination.Study designUncontrolled field trials.MethodsA pilot study evaluated the motility of frozen–thawed spermatozoa after centrifugation and storage (17°C) in two different media for up to 48 h. Subsequently, the motility of frozen–thawed semen processed through a microfluidic device, resuspended in two different media during storage (17°C) for up to 24 h was evaluated. The fertility of frozen–thawed spermatozoa, after microfluidic sorting and storage at 17°C for up to 24 h, was evaluated after fixed‐time insemination in a commercial embryo programme. Experiment 1: Frozen–thawed spermatozoa (N = 5 stallions) were centrifuged and resuspended in Botusemen Gold™ or SpermSafe™ and stored (17°C) for up to 48 h. Sperm motility was evaluated by CASA at 0, 6, 24 and 48 h. Experiment 2: Frozen–thawed spermatozoa (N = 4 stallions) underwent microfluidic sorting and storage (17°C) for up to 24 h in both media. Sperm concentration and motility were evaluated at 0, 16 and 24 h. Experiment 3: Fertility of frozen–thawed spermatozoa (N = 3 stallions) was evaluated after insemination of 42 mare cycles at 6, 16 and 24 h after thawing, microfluidic sorting and storage before fixed‐time insemination.ResultsThe stallion significantly influenced sperm motility, but there was no effect of media on motility parameters. Storage time significantly affected sperm motility after centrifugation but not after microfluidic sorting. Storage time had no effect on the overall embryo recovery rate (52%, n = 42).Main limitationsField trial with small mare numbers and no control at time = 0 h.ConclusionsFixed‐time insemination of frozen–thawed spermatozoa after microfluidic sorting and storage at 17°C for up to 24 h produced satisfactory embryo recovery rates.

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“…The use of cooled semen still constitutes the core of most assisted reproductive technologies (ARTs) for equine breeding. 1 This ART was developed in the second half of the last century and established the basis of currently used commercial extenders, which saw several improvements by the end of the century. When using these extenders, refrigeration is apparently required, a handicap when considering long-haul transport of the semen, but alternatives, such as use of room-temperature storage are scarce.…”

Section: Introductionmentioning

confidence: 99%

Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67 mm glucose–10 mm pyruvate‐based media

Becerro‐Rey,

Martín‐Cano,

Ferrusola

et al. 2023

Andrology

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BackgroundMost commerce of equine seminal doses is carried out using commercial extenders under refrigeration at 5°C.ObjectivesTo determine if 10 mm pyruvate in a 67 mm glucose extender and storage at 22°C could be the basis of an alternative storage method to cooling to 5°C.Material and methodsStallion ejaculates were extendedin: INRA96 (67 mm glucose, non‐pyruvate control), modified Tyrode's (67 mm glucose–10 mm pyruvate), supplemented with 0, 10, 50, and 100 μM itaconate. As itaconate was vehiculated in DMSO, a control vehicle was also included. Sperm motility, viability, mitochondrial membrane potential, and production of reactive oxygen species were measured after collection and again after 48 and 96 h of storage at 22°C. To disclose molecular metabolic changes, spermatozoa were incubated up to 3 h in modified Tyrode's 67 mm glucose–10 mm pyruvate and modified Tyrode's 67 mm glucose, and metabolic analysis conducted.ResultsAfter 96 h of storage aliquots stored in the control, INRA96 had a very poor total motility of 5.6% ± 2.3%, while in the 67 mm glucose–10 mm pyruvate/10 μm itaconate extender, total motility was 34.7% ± 3.8% (p = 0.0066). After 96 h, viability was better in most pyruvate‐based media, and the mitochondrial membrane potential in spermatozoa extended in INRA96 was relatively lower (p < 0.0001). Metabolomics revealed that in the spermatozoa incubated in the high pyruvate media, there was an increase in the relative amounts of NAD+, pyruvate, lactate, and ATP.Discussion and conclusionsAliquots stored in a 67 mm glucose–10 mm pyruvate‐based medium supplemented with 10 μM itaconate, maintained a 35% total motility after 96 h of storage at 22°C, which is considered the minimum acceptable motility for commercialization. Improvements may be related to the conversion of pyruvate to lactate and regeneration of NAD+.

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Liquid storage of stallion spermatozoa – Past, present and future

Cited by 9 publications

References 101 publications

Recent advances in reproductive research in Australia and New Zealand: highlights from the Annual Meeting of the Society for Reproductive Biology, 2022

Recent advances in reproductive research in Australia and New Zealand: highlights from the Annual Meeting of the Society for Reproductive Biology, 2022

A simplified fixed‐time insemination protocol using frozen–thawed stallion spermatozoa stored at 17°C for up to 24 h before insemination

Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67 mm glucose–10 mm pyruvate‐based media

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