Skim milk powder used as a non-permeable cryoprotectant reduces oxidative and DNA damage in cryopreserved zebrafish sperm

Low temperatures during cryopreservation activate a cascade of changes, which may lead into irreversible damage and reduction of the fertilization potential, including the process of premature capacitation. The aim of our study was to evaluate the range of cell damage following the cryopreservation process and possible activation of cryocapacitation in bovine spermatozoa. For the experiments semen samples were obtained from 30 sexually mature Holstein bulls. Within the analysed parameters, we focused on the functional activity, structural integrity, capacitation status and oxidative profile. The samples were divided into three experimental groups, control (CTRL), in vitro capacitated (CAP) and cryopreserved (CRYO). Based on the collected data, there was a significant decrease in the sperm motility, mitochondrial membrane potential and concentration of cyclic adenosine monophosphate in the CRYO group when compared to CAP and CTRL (P<0.0001). A significant decrease (P<0.01; P<0.0001) in the membrane and acrosome integrity as well as DNA fragmentation index and a significant increase (P<0.0001) of necrotic cells were observed in the CRYO group. Following capacitation, a significant increase (P<0.01; P<0.0001) was recorded in the number of cells which underwent the acrosome reaction in the CRYO group against CAP and CTRL. Changes in the oxidative profile of the CRYO group indicates an increase (P<0.0001) in the reactive oxygen species generation, except for the superoxide radical, which was significantly higher (P<0.0001; P<0.001) in the CAP group in comparison with CRYO and CTRL. In summary, premature capacitation may be considered a consequence of cryopreservation and the assessed parameters could serve as physical markers of cryogenic damage to bovine spermatozoa in the future.

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“…morphology during cryopreservation and after thawing. [26,27]. Casein micelles are considered to be a major protective constituent of skim milk.…”

Section: Plos Onementioning

confidence: 99%

In vitro versus cryo-induced capacitation of bovine spermatozoa, part 1: Structural, functional, and oxidative similarities and differences

Benko

Mohammadi‐Sangcheshmeh²,

Ďuračka³

et al. 2022

PLoS ONE

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“…The cell viability levels found in this research agree with those reported by other authors, in relation to the survival of freeze-dried Lactobacillus , who report a range of 86.82–93.98% being the highest values obtained when skim milk powder was used as a cryoprotective agent [ 36 ], which was also used in this research. Among the benefits associated with skim milk as an effective cryoprotectant, it has been noted that it stabilizes cell membrane constituents and prevents cell damage [ 37 ].…”

Section: Resultsmentioning

confidence: 99%

“…Microorganisms 2024, 12, x FOR PEER REVIEW 15 of 2 86.82%-93.98% being the highest values obtained when skim milk powder was used as cryoprotective agent [36], which was also used in this research. Among the benefits asso ciated with skim milk as an effective cryoprotectant, it has been noted that it stabilizes ce membrane constituents and prevents cell damage [37].…”

Section: Effect Of Drying Strategies On Water Activitymentioning

confidence: 99%

Examining the Effect of Freezing Temperatures on the Survival Rate of Micro-Encapsulated Probiotic Lactobacillus acidophilus LA5 Using the Flash Freeze-Drying (FFD) Strategy

Acosta-Piantini,

Villarán,

Martínez

et al. 2024

Microorganisms

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This work proposes a novel drying method suitable for probiotic bacteria, called flash freeze-drying (FFD), which consists of a cyclic variation in pressure (up-down) in a very short time and is applied during primary drying. The effects of three FFD temperatures (−25 °C, −15 °C, and −3 °C) on the bacterial survival and water activity of Lactobacillus acidophilus LA5 (LA), previously microencapsulated with calcium alginate and chitosan, were evaluated. The total process time was 900 min, which is 68.75% less than the usual freeze-drying (FD) time of 2880 min. After FFD, LA treated at −25 °C reached a cell viability of 89.94%, which is 2.74% higher than that obtained by FD, as well as a water activity of 0.0522, which is 55% significantly lower than that observed using FD. Likewise, this freezing temperature showed 64.72% cell viability at the end of storage (28 days/20 °C/34% relative humidity). With the experimental data, a useful mathematical model was developed to obtain the optimal FFD operating parameters to achieve the target water content in the final drying.

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“…Semen cryopreservation is useful to preserve genetic resources (Chelucci et al, 2015). The composition of the extenders is a critical factor to protect sperm during the cryopreservation process (Alcay et al, 2016; Dhami et al, 1996; Rodrigues et al, 2020; Tuncer et al, 2011). Some extenders and their modifications have been developed for the cryopreservation of goat semen.…”

Section: Discussionmentioning

confidence: 99%

Fertility results after exocervical insemination using goat semen cryopreserved with extenders based on egg yolk, skim milk, or soybean lecithin

Liang

Allai

et al. 2022

Reprod Domestic Animals

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Skim milk powder used as a non-permeable cryoprotectant reduces oxidative and DNA damage in cryopreserved zebrafish sperm

Cited by 13 publications

References 52 publications

In vitro versus cryo-induced capacitation of bovine spermatozoa, part 1: Structural, functional, and oxidative similarities and differences

In vitro versus cryo-induced capacitation of bovine spermatozoa, part 1: Structural, functional, and oxidative similarities and differences

Examining the Effect of Freezing Temperatures on the Survival Rate of Micro-Encapsulated Probiotic Lactobacillus acidophilus LA5 Using the Flash Freeze-Drying (FFD) Strategy

Fertility results after exocervical insemination using goat semen cryopreserved with extenders based on egg yolk, skim milk, or soybean lecithin

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