Freezing decreases sperm quality, ultimately affecting fertilizing ability. The repair of freeze-damaged sperm is considered crucial for improving post-thaw viability and fertility. We investigated the effects of exosomes derived from canine adipose-derived mesenchymal stem cells on dog sperm structure and function during cryopreservation. The pooled ejaculate was diluted with buffer, without (Control), or with exosomal proteins (25, 50, or 100 µg/mL). Using fresh semen, the determined optimal exosomal protein concentration was 50 µg/mL (Group 2) which was used in further experiments. Post-thaw sperm treated with exosomes were superior to control (p < 0.05) in terms of motility (56.8 ± 0.3% vs. 47.2 ± 0.3%), live sperm percentage (55.9 ± 0.4% vs. 45.4 ± 0.4%), membrane integrity (55.6 ± 0.5% vs. 47.8 ± 0.3%), and acrosome integrity (60.4 ± 1.1% vs. 48.6 ± 0.4%). Moreover, expression of genes related to the repair of the plasma membrane (ANX 1, FN 1, and DYSF), and chromatin material (H3, and HMGB 1) was statistically higher in exosome-treated sperm than control, but the expression of the mitochondrial reactive oxygen species modulator 1 gene was significantly higher in control. Therefore, exosomal treatment may improve the quality of post-thaw dog semen through initiating damaged sperm repair and decreasing reactive oxygen species production.

Improved Post-Thaw Quality of Canine Semen after Treatment with Exosomes from Conditioned Medium of Adipose-Derived Mesenchymal Stem Cells.

Effects of kinetin supplementation on the post-thaw motility, viability, and structural integrity of dog sperm.

Cold case: Small animal gametes cryobanking.

Cyclodextrins or cholesterol-loaded-cyclodextrins? A better choice for improved cryosurvival of chicken spermatozoa.

Beneficial influence of fetal bovine serum on in vitro cryosurvival of chicken spermatozoa

Neonatal calves are prone to gastrointestinal infections and microbial dysbiosis that lead to high morbidity and mortality. Prebiotics can be used to mitigate the adverse effects of gut diseases and microbial dysbiosis. Forty male Holstein-Friesian calves (2 ± 1 day old) were divided into four dietary treatments: control (milk without prebiotics), YCW-2, YCW-4 (milk containing 2 or 4 g/day/calf of yeast cell wall, respectively), and cMOS (milk containing commercial mannan-oligosaccharides 4 g/day/calf). Milk intake, feed intake, and health scores were recorded daily, whereas body weight, dry matter intake (DMI), and body measurements were recorded weekly. Feed efficiency (FE) was determined at the end of the trial (eight weeks). Cell-mediated immunity (CMI) was assessed by the topical application of dinitrochlorobenzene. Blood samples were collected fortnightly to determine glucose, non-esterified fatty acid (NEFA), blood urea nitrogen (BUN), and beta-hydroxybutyric acid (βHBA). Cell wall supplemented calves had significantly higher (P < 0.05) final body weights, DMI, and body measurements along with improved (P < 0.05) faecal scores. Feed intake was higher (P < 0.05) in both the YCW-supplemented calves. Glucose was lower (P < 0.001), whereas BUN and βHBA were significantly higher in the YCW-2 animals. No differences were observed in FE, CMI, and NEFA between all the experimental animals. The yeast cell wall (2 g) may have the potential to improve the growth performance and health status of neonatal calves.

Prebiotics supplementation modulates pre-weaning stress in male cattle calves by improving growth performance, health scores and serum biomarkers

Contents
Cryopreservation affects integrity of cholesterol and phospholipids in the plasma membrane of sperm leading to decreased fertility of frozen-thawed semen. Cholesterol-loaded cyclodextrins (CLC) have been shown to improve post-thaw semen quality in various species. The aim of this study was to investigate the optimal concentration of CLC for better post-thaw semen quality in dogs. Semen collection, through digital manipulation, was conducted once a week in four adult German shepherd dogs (n = 20 ejaculates; five ejaculates/dog). Semen samples with mass motility>3 (0: without movement; 5: fast progressive movement), motility >70% and concentration >200 × 106/ml were pooled and processed in Tris-citrate extender containing 0, 1, 2 or 3 mg of CLC. The post-thaw quality was assessed on the basis of percentage motility, morphological abnormalities, live/dead ratio and plasma membrane, acrosome and DNA integrity, evaluated using anova and further analysed by Tukey's range test, if applicable. The addition of CLC showed an overall improvement in post-thaw semen quality. Among various treatment groups, and when compared to the control, the percentages of motile (55.5%), viable (65%), plasma membrane intact (56.7%), acrosome intact (49.2%) and DNA intact (98%) spermatozoa were significantly higher in 2 mg/ml CLC group (p < .05). It is concluded that incorporation of cholesterol in semen extender results in a beneficial increase in post-thaw semen quality in dogs.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/rda.12893

Effect of cholesterol-loaded cyclodextrins on cryosurvival of dog spermatozoa.

Effects of high dietary threonine supplementation on growth performance, health biomarkers, and intestinal histology in cyclic heat-stressed broilers

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Prebiotics supplementation modulates pre-weaning stress in male cattle calves by improving growth performance, health scores and serum biomarkers

Effect of cholesterol-loaded cyclodextrins on cryosurvival of dog spermatozoa.

Effects of high dietary threonine supplementation on growth performance, health biomarkers, and intestinal histology in cyclic heat-stressed broilers