New strategies of boar sperm cryopreservation: Development of novel freezing and thawing methods with a focus on the roles of seminal plasma

Some potential markers of boar sperm freezability have been found in spermatozoa, but little attention has been paid to seminal plasma. The seminal plasma is composed of secretions from the testis, epididymis, and accessory sex glands. The exposure of spermatozoa to small molecules such as metabolites can affect sperm function. However, details and significance of the seminal plasma metabolome related to boar sperm freezability are unknown. Therefore, the main aim of this study was to explore the differences in the metabolic level of seminal plasma between boars with differential freezability and to explore the candidate biomarkers of semen freezability. A total of 953 metabolites were identified in boar semen plasma by UHPLC-qTOF-MS analysis, and 50 metabolites showed significant change between the GFE group and PFE group. Further, twelve metabolites were subjected to metabolic target analysis, and three metabolites (D-aspartic acid, N-acetyl-L-glutamate (NAG), and inosine) showed differences. In conclusion, there is significant difference in the metabolome of seminal plasma between GFE and PFE individuals. D-aspartic acid, NAG, and inosine in seminal plasma may be potential markers for assessing sperm cryopreservation resistance in boars.

Download Full-text

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

Animals ◽

10.3390/ani11010203 ◽

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.

Download Full-text

Macro- and microelements in serum and seminal plasma as biomarkers for bull sperm cryotolerance

Acta Veterinaria Scandinavica ◽

10.1186/s13028-021-00590-2 ◽

2021 ◽

Vol 63 (1) ◽

Author(s):

Maja Zakošek Pipan ◽

Petra Zrimšek ◽

Breda Jakovac Strajn ◽

Katarina Pavšič Vrtač ◽

Tanja Knific ◽

...

Keyword(s):

Seminal Plasma ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Quality Characteristics ◽

Freezing And Thawing ◽

Additional Tool ◽

Bull Semen ◽

Bull Sperm ◽

Fresh Semen

ABSTRACT Background Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.

Download Full-text

Seminal plasma arising from the whole boar sperm-rich fraction increases the stability of sperm membrane after thawing1,2

Journal of Animal Science ◽

10.2527/jas.2016-0293 ◽

2016 ◽

Vol 94 (5) ◽

pp. 1906-1912 ◽

Cited By ~ 11

Author(s):

M. A. Torres ◽

G. M. Ravagnani ◽

D. F. Leal ◽

S. M. M. K. Martins ◽

B. B. D. Muro ◽

...

Keyword(s):

Seminal Plasma ◽

Boar Sperm ◽

Sperm Membrane ◽

The Stability

Download Full-text

Absence of seminal plasma from sperm-rich fraction decreases boar sperm quality characteristics during the course of liquid storage

Animal Reproduction Science ◽

10.1016/j.anireprosci.2018.08.029 ◽

2018 ◽

Vol 198 ◽

pp. 20-26 ◽

Cited By ~ 3

Author(s):

D.F. Leal ◽

M.A. Torres ◽

G.M. Ravagnani ◽

S.M.M.K. Martins ◽

F.V. Meirelles ◽

...

Keyword(s):

Seminal Plasma ◽

Sperm Quality ◽

Quality Characteristics ◽

Liquid Storage ◽

Boar Sperm

Download Full-text

In vitro effects of two different commercial freezing and thawing extenders on boar sperm quality

Animal Reproduction Science ◽

10.1016/j.anireprosci.2021.106906 ◽

2021 ◽

pp. 106906

Author(s):

De Andrade AFC ◽

R Grossfeld ◽

RV Knox

Keyword(s):

Sperm Quality ◽

Freezing And Thawing ◽

Boar Sperm ◽

In Vitro Effects

Download Full-text

Sperm Proteome after Interaction with Reproductive Fluids in Porcine: From the Ejaculation to the Fertilization Site

International Journal of Molecular Sciences ◽

10.3390/ijms21176060 ◽

2020 ◽

Vol 21 (17) ◽

pp. 6060 ◽

Cited By ~ 1

Author(s):

Chiara Luongo ◽

Leopoldo González-Brusi ◽

Paula Cots-Rodríguez ◽

Mª José Izquierdo-Rico ◽

Manuel Avilés ◽

...

Keyword(s):

Seminal Plasma ◽

Proteome Analysis ◽

Interacting Proteins ◽

Sperm Transport ◽

Two Fluids ◽

Boar Sperm ◽

Uterine Fluid ◽

Proteome Changes ◽

Oviductal Fluid

Ejaculated sperm are exposed to different environments before encountering the oocyte. However, how the sperm proteome changes during this transit remains unsolved. This study aimed to identify proteomic changes in boar sperm after incubation with male (seminal plasma, SP) and/or female (uterine fluid, UF; and oviductal fluid, OF) reproductive fluids. The following experimental groups were analyzed: (1) SP: sperm + 20% SP; (2) UF: sperm + 20% UF; (3) OF: sperm + 20% OF; (4) SP + UF: sperm + 20% SP + 20% UF; and (5) SP+OF: sperm + 20% SP + 20% OF. The proteome analysis, performed by HPLC-MS/MS, allowed the identification of 265 proteins. A total of 69 proteins were detected in the UF, SP, and SP + UF groups, and 102 proteins in the OF, SP, and SP + OF groups. Our results showed a higher number of proteins when sperm were incubated with only one fluid than when they were co-incubated with two fluids. Additionally, the number of sperm-interacting proteins from the UF group was lower than the OF group. In conclusion, the interaction of sperm with reproductive fluids alters its proteome. The description of sperm-interacting proteins in porcine species after co-incubation with male and/or female reproductive fluids may be useful to understand sperm transport, selection, capacitation, or fertilization phenomena.

Download Full-text