Impact of different dilution techniques on boar sperm quality and sperm distribution of the extended ejaculate

Schulze, Martin; Ammon, Christian; Schaefer, Joel J.; Luther, Anne-Marie; Jung, Markus; Waberski, Dagmar

doi:10.1016/j.anireprosci.2017.05.013

Cited by 20 publications

(10 citation statements)

References 22 publications

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“…Flow cytometry can be used to measure ion indicators, such as calcium [Ca 2+ ] uptake before and after in vitro capacitation, measuring the responsiveness of spermatozoa to the capacitation stimulus bicarbonate. The Fluo-3 AM (Fluo-3), was first used to assess Ca 2+ uptake in the midpiece region of boar spermatozoa by FC over 25 years ago [53] and has been used to determine the response in a capacitating medium containing bicarbonate, compared with a non-capacitating medium [54][55][56][57], as well as to study the function of the CatSper channel in relation to capacitation in boar spermatozoa [58]. Rhod-5N-AM (Rhod5) has a lower calcium affinity than Fluo-3 and cellular localisation of stained Ca 2+ in the acrosomal and post-acrosomal regions [56].…”

Section: Calcium Uptake and Intracellular Phmentioning

confidence: 99%

An update on boar semen assessments by flow cytometry and CASA

Boe-Hansen

Satake

2019

Theriogenology

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In the quest for predicting fertility of an individual, enhancing semen handling, dilution and storage protocols, and understanding the impact of environment and, andrologists have changed their approaches to semen analysis. The technologies used today are fast developing and readily implemented in research. Semen is one of a few naturally occurring monocellular suspensions, so sperm function analysis by flow cytometry (FC) and utilization of fluorochromes is an ideal technique for high throughput, objective and accurate analysis. The complementary use of microscopical assessments by Computer-Assisted Semen Analysis (CASA), where sperm cell parameters can be objectively assessed is equally important. The objectivity and repeatability of these techniques have driven research on the function, identification of heterogeneity and fertility of the ejaculate. The wealth of knowledge obtained from the application of these powerful methods has changed our view of the spermatozoon.Although there is some application of these methods in the industry producing boar semen for artificial insemination (AI) and to eliminate sires of sub-standard semen quality, uptake of advanced methods is still slow. Instruments are becoming cheaper and technically more user friendly. Standardization of methodology and optimization of instrument settings is important for full implementation of these systems, including comparison between labs.This review provides an update on two technologies: flow cytometry and CASA for objective analysis of boar semen quality.

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Section: Calcium Uptake and Intracellular Phmentioning

confidence: 99%

An update on boar semen assessments by flow cytometry and CASA

Boe-Hansen

Satake

2019

Theriogenology

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“…Preliminary studies using commercial membrane protecting semen extenders AndroStar Plus 24 or Androstar Premium extender 25 (Minitüb, Tiefenbach, Germany) indicate that low-temperature storage in absence of antibiotics could be an option for boar semen preservation if maintenance of sperm quality and control of bacterial growth is achieved. Recent research demonstrated that cell alterations in preserved boar semen can be diminished by isothermic one–step dilution at 30–33 °C followed by slow cooling 26 , a minimum sperm concentration of 15 × 10 6 /ml 27 , use of modern commercial semen extenders 28 and motionless semen storage 29 .…”

Section: Introductionmentioning

confidence: 99%

Sperm function in vitro and fertility after antibiotic-free, hypothermic storage of liquid preserved boar semen

Waberski

Luther

Grünther

et al. 2019

Sci Rep

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The role of antibiotics (AB) in semen extenders as a potential contribution to the global antimicrobial resistance threat is emerging. Here, we establish an AB-free hypothermic preservation strategy for boar semen and investigate its impact on sperm function, microbial load and fertility after artificial insemination (AI). Spermatozoa (12 boars) preserved in AB-free AndroStar Premium extender at 5 °C maintained high motility, membrane integrity, and a low DNA-fragmentation index throughout 72 h storage and results did not significantly differ from controls stored at 17 °C in extender containing AB (p = 0.072). Likewise, kinetic response of spermatoza to the capacitation stimulus bicarbonate during 180 min incubation in Tyrode’s medium did not differ from 17 °C-controls. In a competitive sperm oviduct binding assay, binding indices did not differ between semen stored for 72 h AB-free at 5 °C and 17 °C-controls (n = 6 boars). Bacterial load < 103 CFU/ml after 72 h was measured in 88.9% of samples stored at 5 °C AB-free compared to 97.2% in 17 °C-controls (n = 36 semen pools, 23 boars). Fertility traits of 817 females did not differ significantly between the two semen groups (p > 0.05). In conclusion, a hypothermic semen preservation strategy is presented which offers antibiotic-free storage of boar semen doses.

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“…This indicates that the sperm of purebred breeders are more sensitive to the environmental factors to which semen is exposed after it is collected. Many studies have shown that various exogenous factors can affect the quality of boar sperm during the stages of laboratory processing [ 2 , 9 ] and during storage [ 10 , 20 , 25 ]. Therefore, the sperm of crossbred boars appear to show greater resistance than the sperm of boars of the parent breeds.…”

Section: Discussionmentioning

confidence: 99%

“…The integrity and normal functioning of the sperm cell membrane are essential for sperm metabolism, capacitation, acrosome reaction, and the ability to fertilize the ovum [ 7 ]. Factors such as the rate of cooling [ 8 ], the dilution procedure [ 9 ], and storage conditions [ 10 , 11 ] can adversely affect the structure of the sperm cell membrane. Cooling induces changes in the sperm cell membrane, impairing its functional and molecular state [ 2 , 8 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

Integrity of Sperm Cell Membrane in the Semen of Crossbred and Purebred Boars during Storage at 17 °C: Heterosis Effects

Wysokińska

Szablicka

2021

Animals

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The aim of the study was to assess changes in the integrity of sperm cell membranes during the storage of semen collected from Duroc × Pietrain crossbred boars and purebred boars of the component breeds. To compare the cell membrane integrity of sperm heads in crossbred and purebred boars, heterosis effects were estimated. The study was conducted on 48 ejaculates collected from Duroc × Pietrain crossbred boars and from purebred Duroc and Pietrain boars used for artificial insemination. Microscope slides were prepared from each ejaculate for the evaluation of the cell membrane integrity of the sperm, at 1, 24, 48, 72, and 96 h after collection of the ejaculate. Diluted ejaculates were stored at 17 °C. Sperm membrane integrity was analysed by two methods: SYBR-14/PI and eosin–nigrosin. Our results showed that the cell membrane integrity of sperm heads changed with storage time, but the extent of the changes varied depending on the genetic group of boars. The semen of Duroc × Pietrain crossbreds was clearly seen to be less sensitive to storage conditions than that of boars of the parent breeds, which was confirmed by the calculated heterosis effects. The percentage of sperm with an intact cell membrane was higher in crossbred boars than in purebred boars (p ≤ 0.05). In addition, significantly fewer moribund sperm spermatozoa and spermatozoa with a damaged cell membrane were observed in crossbred boars (p ≤ 0.05). In the semen of purebred Duroc and Pietrain boars, the cell membrane integrity of the sperm should be assessed more often during storage than in the semen of Duroc × Pietrain crossbred boars. This study provides valuable information for the development and implementation of semen quality monitoring in crossbred boars and boars of the parent breeds during storage at 17 °C with respect to the cell membrane structure of sperm heads. The evaluation methods used effectively identify damage to the cell membranes of the sperm during semen storage.

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Impact of different dilution techniques on boar sperm quality and sperm distribution of the extended ejaculate

Cited by 20 publications

References 22 publications

An update on boar semen assessments by flow cytometry and CASA

An update on boar semen assessments by flow cytometry and CASA

Sperm function in vitro and fertility after antibiotic-free, hypothermic storage of liquid preserved boar semen

Integrity of Sperm Cell Membrane in the Semen of Crossbred and Purebred Boars during Storage at 17 °C: Heterosis Effects

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