Slow cooling prevents cold-induced damage to sperm motility and acrosomal integrity in the black-footed ferret (Mustela nigripes)

2007 ◽  
Vol 19 (5) ◽  
pp. 652 ◽  
Author(s):  
R. M. Santymire ◽  
P. E. Marinari ◽  
J. S. Kreeger ◽  
D. E. Wildt ◽  
J. G. Howard
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Artificial Insemination ◽  
Sperm Cryopreservation ◽  
Slow Cooling ◽  
Rapid Cooling ◽  
Mustela Nigripes ◽  
Factors Influencing ◽  
Rate Of Cooling ◽  
Acrosomal Integrity

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) at 25°C than at 37°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°C min–1) was optimal (P < 0.05) compared to rapid cooling (1°C min–1), and ultra-rapid cooling (9°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.

scholarly journals Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda

Reproduction ◽  
2004 ◽  
Vol 127 (5) ◽  
pp. 547-556 ◽  
Author(s):  
R E Spindler ◽  
Y Huang ◽  
J G Howard ◽  
P Wang ◽  
H Zhang ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Giant Panda ◽  
Calcium Ionophore ◽  
Sperm Cryopreservation ◽  
Management Tools ◽  
Giant Pandas ◽  
Before And After ◽  
Cryopreservation Protocol ◽  
Acrosomal Integrity

Sperm cryopreservation and artificial insemination are important management tools for giant panda breeding and the preservation of extant genetic diversity. This study examined the influence of freeze–thawing on sperm function, specifically capacitation. Sperm from nine giant pandas were assessed before and after rapid (− 40 and − 100 °C/min) cryopreservation by incubation in HEPES-buffered Ham’s F10 medium with and without the capacitation accelerators, 3-isobutyl-1-methylxanthine (IBMX) and dibutyryl cyclic AMP (dbcAMP). At 0, 3 and 6 h of exposure, aliquots were assessed for sperm motility traits and capacitation, defined as the proportion of sperm with intact acrosomes following exposure to solubilised zonae pellucidae (ursid or felid) or calcium ionophore subtracted from the proportion of sperm with intact acrosomes before exposure. Although mean±s.e.m. sperm motility post-thaw (56.1 ± 3.9% at 0 h) was less (P < 0.05) than pre-freeze (71.7 ± 6.0%), there was no difference (P > 0.05) in the proportion of acrosome-intact sperm (fresh, 93.0 ± 1.7% versus cryopreserved–thawed, 81.7 ± 4.7% at 0 h). Incidence of capacitation was greater (P < 0.05) in fresh sperm incubated with capacitation accelerators IBMX and dbcAMP (9 h: 50.9 ± 1.1) compared with fresh sperm incubated without accelerators (9 h: 41.2 ± 1.1%). Frozen–thawed sperm preincubated without accelerators underwent capacitation (49.6 ± 1.1%) to a greater extent (P < 0.05) compared with these fresh counterparts. Thawed samples with (9 h: 45.9 ± 1.4%) and without accelerators (9 h: 41.2 ± 1.1%) did not differ (P > 0.05) during the 9-h incubation. We conclude that giant panda spermatozoa (1) undergo capacitation in vitro with or without chemical accelerators and (2) withstand a rapid cryopreservation protocol, including retaining normal acrosomal integrity and functional capacitation ability.

Assessment of basic seminal characteristics, sperm cryopreservation and heterologous in vitro fertilisation in the fishing cat (Prionailurus viverrinus)

2006 ◽  
Vol 18 (3) ◽  
pp. 373 ◽  
Author(s):  
Khongsak Thiangtum ◽  
William F. Swanson ◽  
JoGayle Howard ◽  
Wanchai Tunwattana ◽  
Dakara Tongthainan ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Morphology ◽  
Domestic Cat ◽  
Ex Situ ◽  
In Vitro Fertilisation ◽  
Genetic Management ◽  
Slow Cooling ◽  
Breeding Programmes ◽  
Acrosomal Integrity

Conservation of the fishing cat, a threatened south-east Asian felid, could benefit from effective ex situ genetic management and breeding programmes, including the use of assisted reproduction. The aims of the present study were to: (1) characterise basal seminal traits of fishing cats in Thailand zoos; and (2) investigate the effect of cryopreservation on sperm motility, acrosomal integrity and in vitro function. Seminal traits were evaluated in electroejaculates collected from eight males. Spermatozoa were diluted in n-tris(hydroxymethyl)-methyl-2-aminoethanesulfonic acid Tris (TEST)-yolk buffer (TYB) without glycerol, then diluted further with TYB with glycerol (4% final concentration) at either 25°C or after slow cooling to 5°C and frozen in straws over liquid nitrogen vapour. After thawing, sperm function was assessed by insemination of viable domestic cat oocytes. Fishing cat ejaculates averaged (± s.e.m.) 43.6 ± 14.2 × 106 motile spermatozoa with 33.5 ± 6.8% normal sperm morphology. Semen processing had a negligible effect (P > 0.05) on sperm motility and acrosomal integrity, but values were reduced (P < 0.05) after thawing. All thawed samples fertilised domestic cat oocytes, with 62.1% (36/58) of mature oocytes cleaving. Glycerol addition at 5°C resulted in higher (P < 0.05) post-thaw motility and intact acrosomes than glycerol addition at 25°C. In conclusion, good-quality ejaculates can be obtained from Thai fishing cats and their spermatozoa exhibit adequate function after cryopreservation for in vitro fertilisation procedures.

116 IMPROVED SURVIVAL BY CRYOPRESERVING RHESUS MACAQUE (MACACA MULATTA) SPERMATOZOA WITH DIRECTIONAL FREEZING TECHNIQUE

2010 ◽  
Vol 22 (1) ◽  
pp. 217 ◽  
Author(s):  
W. Si ◽  
Y. Lu ◽  
X. He ◽  
S. Ji ◽  
Y. Niu ◽  
...  
Keyword(s):  
Rhesus Monkey ◽  
Cooling Rate ◽  
Sperm Motility ◽  
Genetically Engineered ◽  
Human Diseases ◽  
Slow Cooling ◽  
Cooling Rates ◽  
Important Species ◽  
Directional Freezing ◽  
Acrosomal Integrity

A significant increase in nonhuman primate models of human diseases will be expected in the near future since the successes in production of genetically engineered rhesus monkey models of human diseases. Sperm banking can provide an effective way to preserve valuable genetic resources. Our objective was to (1) develop a protocol using directional freezing technique (DFT) for rhesus monkey spermatozoa cryopreservation, which allows precise control of the velocity and the morphology of the ice-front propagation by transferring the tubes loaded with 2 mL sperm samples at a controllable velocity through two separate chambers with controllable temperature settings, and (2) achieve survival rate that was higher than that achieved with conventional freezing technique (CFT), by which sperm samples were cryopreserved in 0.25 mL straws with liquid nitrogen vapor in a styrofoam box. Sperm motility, acrosomal integrity, and in vitro fertilization (IVF) assay were used to assess the function of frozen-thawed spermatozoa. Data were analyzed by ANOVA and Fisher protected LSD test. Experiment 1 was aimed at optimizing the cooling rate using DFT. Tubes were frozen using the multi-thermal gradient freezing device (MTG 516, Harmony CryoCareTM, IMT Ltd.) at fast (16°C/min), medium (12°C/min), and slow (7°C/min) cooling rates, which corresponded to the transferring velocities (2.5, 1.5, and 0.5 mm s-1, respectively). The results showed that spermatozoa frozen at fast and medium cooling rates showed significantly higher frozen-thawed motility than those frozen at slow cooling rate (61% and 59% v. 50%, P < 0.05). However, no difference was observed on sperm acrosomal integrity among the experimental groups (84, 80, and 78%, respectively, P > 0.05). The purposes of Experiment 2 were determined to examine if using DFT at the optimized cooling rate (12°C/min) can improve the cryo-survival of rhesus monkey spermatozoa compared with CFT. Our results showed that spermatozoa cryopreserved by using DFT achieved significantly higher frozen-thawed sperm motility that those cryopreserved by using CFT (64 v. 54%, P < 0.05). However, no difference was observed on acrosomal integrity between spermatozoa cryopreserved by DFT and CFT (84 and 83%, respectively; P > 0.05). The function of spermatozoa cryopreserved by using DFT was further evaluated by IVF. Females were treated with rhFSH twice-daily for 8 days after the onset of menses and following a treatment of hCG injection on Day 9. Cumulus-oocyte complexes were collected by laparoscopic follicular aspiration 32 h later. Of the inseminated oocytes, 79% were fertilized and 90 and 53% of the resulting zygotes developed into 2-cell and blastocysts, respectively. The fertilization rate was lower and the blastocyst rate was slightly higher than our previous report when fresh spermatozoa were used for IVF (94 and 52%, respectively). Our results indicate that spermatozoa of rhesus monkeys can be effectively cryopreserved using DFT in large volume. This finding provided a new and effective way for genetics preservation purposes in this important species.

scholarly journals Measurement of Oxidative Stress Index in Seminal Plasma Can Predict In Vivo Fertility of Liquid-Stored Porcine Artificial Insemination Semen Doses

Antioxidants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1203
Author(s):  
Isabel Barranco ◽  
Camila P. Rubio ◽  
Asta Tvarijonaviciute ◽  
Heriberto Rodriguez-Martinez ◽  
Jordi Roca
Keyword(s):  
Oxidative Stress ◽  
Litter Size ◽  
Sperm Motility ◽  
Seminal Plasma ◽  
Artificial Insemination ◽  
Stress Index ◽  
Trolox Equivalent Antioxidant Capacity ◽  
Oxidative Stress Index ◽  
Farrowing Rate

The study evaluated the relation between the oxidative stress index (OSI) in porcine seminal plasma (n = 76) with sperm resilience and in vivo fertility (farrowing rate and litter size of 3137 inseminated sows) of liquid-stored artificial insemination (AI) semen doses. The OSI was assessed as the ratio of advanced oxidation protein products to Trolox-equivalent antioxidant capacity, both measured using an automated analyzer. Sperm motility (computer-assisted sperm analyzer) and viability (flow cytometry) were evaluated in semen AI-doses at 0 and 72 h of storage at 17 °C. Sperm resilience was defined as the difference between storage intervals. Semen AI-doses were hierarchically clustered as having high, medium and low seminal OSI (p < 0.001) with those of low displaying higher resilience (p < 0.01). Boars were hierarchically clustered into two groups (p < 0.001) as having either positive or negative farrowing rate and litter size deviation; the negative one showing higher seminal OSI (p < 0.05). In sum, seminal OSI was negatively related to sperm motility and the in vivo fertility of liquid-stored boar semen AI-doses, with the receiver operating characteristic curve presenting seminal OSI as a good predictive biomarker of in vivo fertility of AI-boars (area under the curve: 0.815, p < 0.05).

Conservation of fresh ram spermatozoa at 5°C in the presence of seminal plasma

2003 ◽  
Vol 83 (2) ◽  
pp. 221-227 ◽  
Author(s):  
A. Morrier ◽  
F. Castonguay ◽  
J. L. Bailey
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Artificial Insemination ◽  
Genital Tract ◽  
Female Genital Tract ◽  
Sperm Function ◽  
Oviduct Fluid ◽  
Ram Sperm ◽  
Female Genital

Seminal plasma aids sperm transport and contains factors beneficial for sperm function. In artificial insemination, however, diluting the semen reduces the concentration of seminal plasma. To test the hypothesis that supplemental seminal plasma in extended ram semen improves conservation at 5°C, we added various concentrations of seminal plasma to semen during storage, and investigated subsequent sperm function in vitro. Semen was divided into three aliquots, extended in a commercial diluent (Triladyl) supplemented with 0, 10 or 25% (vol:vol) ovine seminal plasma and cooled to 5°C. After 8 and 24 h at 5°C, sperm were suspended in a modified synthetic oviduct fluid (SOF-m) at 39°C to mimic the female genital tract at insemination. Sperm aliquots were assessed for motility and chlortetracycline fluorescence after 0, 4 and 8 h in the SOFm. No significant differences were observed due to seminal plasma supplementation during conservation at 5°C or incubation in SOF-m at 39°C. However, decreased sperm motility and fewer non-capacitated sperm were observed concomitant with an augmentation of capacitated and acrosome-reacted cells during incubation in SOF-m. Therefore, the hypothesis that diluent supplementation with homologous seminal plasma improves ram sperm conservation or subsequent sperm function was not supported. Key words: Ovine, ram, sperm, motility, viability, chlortetracycline fluorescence, artificial insemination, SOF

scholarly journals Sperm Cryopreservation in American Flamingo (Phoenicopterus Ruber): Influence of Cryoprotectants and Seminal Plasma Removal

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 203
Author(s):  
María Gemma Millán de la Blanca ◽  
Eva Martínez-Nevado ◽  
Cristina Castaño ◽  
Juncal García ◽  
Berenice Bernal ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Genetic Material ◽  
First Year ◽  
Sperm Cryopreservation ◽  
Straight Line ◽  
The Family ◽  
Second Year ◽  
American Flamingo ◽  
Phoenicopterus Ruber

The American flamingo is a useful model for the development of successful semen cryopreservation procedures to be applied to threatened related species from the family Phoenicopteridae, and to permit genetic material banking. Current study sought to develop effective sperm cryopreservation protocols through examining the influences of two permeating cryoprotectants and the seminal plasma removal. During two consecutive years (April), semen samples were collected and frozen from American flamingos. In the first year, the effect of two permeating cryoprotectants, DMA (dimethylacetamide) (6%) or Me2SO (dimethylsulphoxide) (8%), on frozen–thawed sperm variables were compared in 21 males. No differences were seen between DMA and Me2SO for sperm motility, sperm viability, and DNA fragmentation after thawing. In the second year, the role of seminal plasma on sperm cryoresistance was investigated in 31 flamingos. Sperm samples were cryopreserved with and without seminal plasma, using Me2SO (8%) as a cryoprotectant. The results showed that samples with seminal plasma had higher values than samples without seminal plasma for the following sperm variables: Straight line velocity (22.40 µm/s vs. 16.64 µm/s), wobble (75.83% vs. 69.40%), (p < 0.05), linearity (62.73% vs. 52.01%) and straightness (82.38% vs. 73.79%) (p < 0.01); but acrosome integrity was lower (55.56% vs. 66.88%) (p < 0.05). The cryoresistance ratio (CR) was greater in samples frozen with seminal plasma than without seminal plasma for CR-progressive motility (138.72 vs. 54.59), CR-curvilinear velocity (105.98 vs. 89.32), CR-straight line velocity (152.77 vs. 112.58), CR-average path velocity (122.48 vs. 98.12), CR-wobble (111.75 vs. 102.04) (p < 0.05), CR-linearity (139.41 vs. 113.18), and CR-straightness (124.02 vs. 109.97) (p < 0.01). This research demonstrated that there were not differences between Me2SO and DMA to successful freezing sperm of flamingos; seminal plasma removal did not provide a benefit for sperm cryopreservation.

scholarly journals The characteristics of proteome and metabolome associated with contrasting sperm motility in goat seminal plasma

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Baoyu Jia ◽  
Jiachong Liang ◽  
Chunrong Lv ◽  
Sameeullah Memon ◽  
Yi Fang ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Enrichment Analysis ◽  
Salivary Secretion ◽  
Metabolic Process ◽  
Functional Roles ◽  
Kegg Analysis ◽  
Parallel Reaction Monitoring ◽  
Metabolite Profiles ◽  
Close Relationship

AbstractSperm motility is an index tightly associated with male fertility. A close relationship between seminal plasma and sperm motility has been confirmed. This study was to assess the protein and metabolite profiles of seminal plasma obtained from adult goats with high or low sperm motility using the proteomic and metabolomic strategies. In total, 2098 proteins were found. 449 differentially abundant proteins (DAPs) were identified, and 175 DAPs were enriched in the high motility group. The obtained DAPs primarily exist in cytoplasma and extra-cellular portion. The Gene Ontology enrichment analysis demonstrated the main functional roles of these DAPs in regulating biological process, metabolic process of organic substances, cellular-metabolic process, primary-metabolic process, metabolic process of nitrogen compounds, etc. Additionally, the Kyoto-Encyclopedia of Genes and Genomes (KEGG) analysis revealed that these DAPs were primarily involved in phosphatidylinositol signaling system, salivary secretion, proteasome, apoptosis, mitophagy-animal, etc. Aided by the parallel reaction monitoring technology, the abundance changing pattern of 19 selected DAPs was consistent with that of the corresponding proteins obtained by TMT. A total of 4603 metabolites were identified in seminal plasma. 1857 differential metabolites were found between the high motility group and the low motility group, and 999 metabolites were up-regulated in the high motility group. The KEGG analysis demonstrated the primary involvement of the differential metabolites in metabolic and synthetic activities. In conclusion, we first established the proteome and metabolome databank of goat seminal plasma, detecting some proteins and metabolites which may affect sperm motility. This study will be valuable for understanding mechanisms leading to poor sperm motility.

scholarly journals Specific Seminal Plasma Fractions Are Responsible for the Modulation of Sperm–PMN Binding in the Donkey

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1388
Author(s):  
Jordi Miró ◽  
Jaime Catalán ◽  
Henar Marín ◽  
Iván Yánez-Ortiz ◽  
Marc Yeste
Keyword(s):  
Molecular Weight ◽  
Sperm Motility ◽  
Seminal Plasma ◽  
Potential Effect ◽  
Polymorphonuclear Neutrophils ◽  
Low Fertility ◽  
Plasma Fractions ◽  
Complex Fluid ◽  
First Time

While artificial insemination (AI) with frozen-thawed sperm results in low fertility rates in donkeys, the addition of seminal plasma, removed during cryopreservation, partially counteracts that reduction. Related to this, an apparent inflammatory reaction in jennies is induced following AI with frozen-thawed sperm, as a high amount of polymorphonuclear neutrophils (PMN) are observed within the donkey uterus six hours after AI. While PMN appear to select the sperm that ultimately reach the oviduct, two mechanisms, phagocytosis and NETosis, have been purported to be involved in that clearance. Remarkably, sperm interacts with PMN, but the presence of seminal plasma reduces that binding. As seminal plasma is a complex fluid made up of different molecules, including proteins, this study aimed to evaluate how different seminal plasma fractions, separated by molecular weight (<3, 3–10, 10–30, 30–50, 50–100, and >100 kDa), affect sperm–PMN binding. Sperm motility, viability, and sperm–PMN binding were evaluated after 0 h, 1 h, 2 h, 3 h, and 4 h of co-incubation at 38 °C. Two seminal plasma fractions, including 30–50 kDa or 50–100 kDa proteins, showed the highest sperm motility and viability. As viability of sperm not bound to PMN after 3 h of incubation was the highest in the presence of 30–50 and 50–100 kDa proteins, we suggest that both fractions are involved in the control of the jenny’s post-breeding inflammatory response. In conclusion, this study has shown for the first time that specific fractions rather than the entire seminal plasma modulate sperm–PMN binding within the donkey uterus. As several proteins suggested to be involved in the control of post-AI endometritis have a molecular weight between 30 and 100 kDa, further studies aimed at determining the identity of these molecules and evaluating their potential effect in vivo are much warranted.

Cooled semen for fixed-time artificial insemination in beef cattle

2016 ◽  
Vol 28 (7) ◽  
pp. 1004 ◽  
Author(s):  
Juliana C. Borges-Silva ◽  
Márcio R. Silva ◽  
Daniel B. Marinho ◽  
Eriklis Nogueira ◽  
Deiler C. Sampaio ◽  
...  
Keyword(s):  
Beef Cattle ◽  
Sperm Motility ◽  
Artificial Insemination ◽  
Egg Yolk ◽  
Fixed Time ◽  
Pregnancy Rates ◽  
Reproductive Efficiency ◽  
Sperm Abnormalities ◽  
Hypoosmotic Swelling Test ◽  
Interesting Approach

This study evaluated the use of cooled semen in a fixed-time artificial insemination (FTAI) program compared with frozen–thawed semen to improve pregnancy rates in beef cattle. Ejaculates of three bulls were collected and divided into two treatments: (1) frozen–thawed semen and (2) cooled semen. Egg-yolk extender without glycerol was used for the cooled semen treatment. Straws (25 × 106 spermatozoa) were submitted to cooling for preservation at 5°C for 24 h, after which FTAI was performed. Nelore cows (n = 838) submitted to FTAI were randomly inseminated using frozen–thawed semen or cooled semen. There was a 20% increase in the pregnancy per AI (P AI–1) using cooled semen compared with frozen–thawed semen (59.9 ± 4.7 vs 49.4 ± 5.0%; P < 0.005). There was no difference in P AI–1 among the bulls (P = 0.40). The frozen–thawed semen had fewer functional spermatozoa than did the cooled semen when evaluated by sperm motility (61.7 vs 81.0%), slow thermoresistance test (41.7 vs 66.7%) and hypoosmotic swelling test (38.3 vs 53.7%; P < 0.05). The percentage of sperm abnormalities did not differ between the freeze–thawing and cooling processes (18.6 vs 22.1%; P > 0.05). Because there was less damage to spermatozoa and improvement in P AI–1, the use of cooled semen instead of frozen–thawed semen is an interesting approach to increase reproductive efficiency in cattle submitted to a FTAI protocol.

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