Quality assessment of cryopreserved New Zealand white rabbit spermatozoa in INRA-82 extender containing different cryoprotectants

Fadl, Aya M.; Ghallab, A. M.; Abou-Ahmed, M.M.

doi:10.4995/wrs.2019.10892

Cited by 10 publications

(12 citation statements)

References 27 publications

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“…DNA integrity is an essential indicator of the quality of cryopreserved sperm (Berlinguer et al., 2009). Harmful effects of freezing and thawing on sperm DNA integrity have been reported in rabbits (Fadl et al., 2019), rams (Succu et al., 2011), boars (Fraser & Strzezek, 2005) and humans (Donnelly et al., 2001). Interestingly, here we found that the addition of melatonin to cooling and freezing extenders could preserve rabbit spermatozoa's DNA integrity (Tables 3 and 4), with 1.0 mM being the most effective concentration.…”

Section: Discussionmentioning

confidence: 99%

“…The functional integrity of the sperm plasma membrane was evaluated by the osmotic resistance test, as described previously (Fadl et al., 2019). Briefly, a 10 μl aliquot of cooled or frozen/thawed semen sample was mixed with 40 μl of distilled water and incubated for 5 min at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…Semen was collected from eight healthy mature (age of 10 months; 4–6 kg body weight) individually housed in flat‐deck wire cages New Zealand white rabbit bucks, exposed to a 16 hr light/8 hr dark photoperiod and temperatures ranging from 19 to 25°C. All animals were kept on a farm belonging to the Faculty of Veterinary Medicine, Cairo University, and were fed ad libitum on a standard commercial diet, and water was supplied continuously (Fadl et al., 2019). Semen was collected regularly (two ejaculates/buck/week) from each buck early in the morning using a lubricated and pre‐warmed (42–45°C) artificial vagina (All Vet.…”

Section: Methodsmentioning

confidence: 99%

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Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa

Fadl

Ghallab

Abou-Ahmed

et al. 2020

Reprod Domestic Animals

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Melatonin is known to protect sperm against freezing‐inflicted damage in different domestic species. The aim of the study was to evaluate the effect of supplementation of semen extender with melatonin on the quality and DNA integrity of cooled and frozen/thawed rabbit spermatozoa. We also investigated whether the addition of melatonin to the semen extender could improve the fertility of rabbit does artificially inseminated with frozen/thawed semen. Semen samples collected from eight rabbit bucks were pooled and then diluted in INRA‐82 supplemented either with (0.5, 1.0 or 1.5 mM) or without (0.0 mM) melatonin. Diluted semen was cooled at 5°C for 24 hr. For cryopreservation and based on the first experiment's best result, semen samples were diluted in INRA‐82 in the presence or absence of 1.0 mM melatonin and then frozen in 0.25 ml straws. Following cooling or thawing, sperm quality and DNA integrity were evaluated. Furthermore, the fertility of frozen/thawed semen was investigated after artificial insemination. Supplementation of semen extender with 1.0 mM melatonin improved (p < .05) motility, viability, membrane and acrosome integrities in cooled semen compared with other groups. Sperm quality and DNA integrity were higher (p < .05) in frozen/thawed semen diluted in 1.0 mM melatonin‐supplemented extender than in the control group. Conception and birth rates were higher in does inseminated with 1.0 mM melatonin treated semen compared with the controls. In conclusion, supplementation of semen extender with 1.0 mM melatonin improved the quality of cooled and frozen/thawed rabbit spermatozoa. Melatonin can preserve DNA integrity and enhance the fertility of frozen/thawed rabbit spermatozoa.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa

Fadl

Ghallab

Abou-Ahmed

et al. 2020

Reprod Domestic Animals

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“…Sperm progressive motility was evaluated at 24, 36, and 48 h post-cooling according to the method described previously (Fadl et al, 2019). Briefly, a small drop of chilled semen was placed onto a pre-warmed glass slide and covered with a coverslip.…”

Section: Evaluation Of Sperm Motilitymentioning

confidence: 99%

Comparison between Tris-buffer and INRA-82 extenders on the quality of chilled rabbit spermatozoa

2020

Self Cite

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The aim of the present study was to compare the effects of tris-buffer and INRA-82 extenders on the quality of cooled rabbit spermatozoa. Pooled semen samples were collected from eight New Zealand White rabbit bucks, divided into three groups and diluted 1:5 with three different semen extenders: INRA-82, tris-citrate glucose (TCG) and tris-citrate trehalose (TCT). Following dilution, the samples were stored at 5°C for up to 48 h. Sperm motility was recorded at 24, 36 and 48 h post-cooling. Furthermore, sperm viability, morphology, membrane integrity, acrosome status and DNA integrity were assessed at 24 h post-cooling. We found that progressive motility percentages were significantly higher (P<0.05) in samples diluted in INRA-82 (61.00, 53.50, 44.00% at 24, 36 and 48 h post-cooling, respectively) than those diluted in either TCG (39.25, 32.00, 19.75%) or in TCT (47.25, 40.50, 29.00%). We also reported that sperm viability, percentage of normal spermatozoa, percentage of spermatozoa with intact acrosome and DNA integrity after 24 h cooling were significantly higher (P<0.05) in INRA-82 diluted samples compared to TCG and TCT diluted ones. In summary, our results show that dilution of rabbit semen in INRA-82 improved sperm physiological parameters post-cooling compared to those diluted in TCG or TCT. Our findings also suggest that INRA-82 is a promising diluent that can be used effectively to maintain the viability of chilled rabbit semen.

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“…Pooled semen samples were evaluated for basic semen characteristics. Pooled semen samples were diluted 1:1 (V/V) with INRA-82 extender composed of glucose 25 g/L, lactose 1.5 g/L, raffinose 1.5 g/L, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid 4.76 g/L, potassium citrate 0.41 g/L and sodium citrate 0.25 g/L, 0.15% skimmed milk, 20% egg yolk (Vidament et al, 2000) and supplemented with dimethyl sulfoxide 4%+4% as a cryoprotectant (Fadl et al, 2019). Diluted samples were then kept in the refrigerator at 5°C (in graduated Falcon tubes).…”

Section: Semen Collection and Processingmentioning

confidence: 99%

Effect of seasonality on quality and fertility of cryopreserved New Zealand white rabbit semen under Egyptian conditions

Fadl

2020

World Rabbit Sci.

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The present study was designed to investigate the effect of seasonality on the quality and fertility of cryopreserved New Zealand white (NZW) rabbit semen under Egyptian conditions. Semen was collected from fifteen mature rabbit bucks during winter, spring, summer and autumn seasons, diluted in INRA-82 extender and cryopreserved in liquid nitrogen. Following the freezing/thawing process, semen samples were evaluated for semen quality parameters. We also investigated whether seasonality could affect postthaw fertility. The results showed that semen collected and processed during winter, autumn, and spring significantly improved (P<0.05) sperm motility (56.33, 54.12 and 54.00%, respectively), live sperm (59.24, 58.34 and 57.01%, respectively), functional membrane integrity hypo osmotic swelling test (67.34, 64.59 and 64.31%, respectively), acrosome intactness (79.25, 77.45 and 76.72%, respectively) and sperm with non-fragmented DNA (75.66, 74.02% and 72.53%, respectively) in comparison with semen collected and processed in summer season (30.24, 33.26, 42.51, 51.20 and 40.22%, respectively). The fertility results revealed that conception and kindling rates were significantly higher (P<0.05) when semen samples were collected and processed in winter (80.00 and 76.00%, respectively) in comparison with those collected and processed in autumn (74.00 and 70.00%, respectively), spring (70.00 and 64.00%, respectively) and summer (30.00 and 20.00%, respectively). In summary, our results show that the quality and fertility of cryopreserved NZW rabbit buck semen depends on the season during which the semen is collected and processed. Moreover, for the best fertility results in doe insemination, semen collected and processed (cryopreserved) in winter season should be used.

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Quality assessment of cryopreserved New Zealand white rabbit spermatozoa in INRA-82 extender containing different cryoprotectants

Cited by 10 publications

References 27 publications

Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa

Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa

Comparison between Tris-buffer and INRA-82 extenders on the quality of chilled rabbit spermatozoa

Effect of seasonality on quality and fertility of cryopreserved New Zealand white rabbit semen under Egyptian conditions

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