Effect of cryopreservation and single layer centrifugation on canine sperm DNA fragmentation assessed by the sperm chromatin dispersion test

DNA integrity of fresh and frozen canine epididymal spermatozoa

“…The SCD test was reported to be simple, highly reproducible and inexpensive technique, and the SCD results correlated with the SCSA results [28]. In the current paper, the DNA fragmentation indices were similar in fresh and frozen canine epididymal spermatozoa, which confirms the resilience of canine ejaculated sperm DNA to cold stress [12,15,16]. A progressive deterioration of DNA has been observed after exposure of fresh or thawed spermatozoa to different stressors (e.g., incubation time and temperature, toxicants, ROS) [21,29], indicating that thefreezing/thawing procedure facilitates the destabilization of the chromatin structure of spermatozoa [21].…”

“…The impact of cryopreservation on sperm DNA integrity is still a controversial matter in mammals including dogs. Some authors showed that the freezing/thawing procedure did not produce significant adverse effects on chromatin status in canine ejaculated spermatozoa [12,15,16]. In contrast, Kim et al [14] found a higher DNA fragmentation in thawed compared to fresh spermatozoa.…”

“…In contrast, Kim et al [14] found a higher DNA fragmentation in thawed compared to fresh spermatozoa. In the present study, DNA fragmentation was examined with the use of a commercial kit Halomax ® based on the sperm chromatin dispersion (SCD) test -previously employed for ejaculated canine semen [9,16], whereas the cited authors used the sperm chromatin structure Data derived from 10 replicates; different superscripts within a row indicate significant differences (p < 0.0001). assay (SCSA) and acridine orange assay.…”

“…Sperm DNA integrity has been evaluated in fresh ejaculated [7][8][9][10] and epididymal canine semen [11]. Few reports have compared fresh and post-thaw chromatin integrity of canine ejaculated spermatozoa, obtaining variable results [12][13][14][15][16], but the post-thaw DNA stability of canine epididymal spermatozoa has not been investigated. The integrity of the paternal DNA is of crucial importance for embryo development [17], and a relationship between DNA damage and infertility has been demonstrated in humans.…”

DNA integrity of canine spermatozoa during chill storage assessed by the sperm chromatin dispersion test using bright-field or fluorescence microscopy

“…There are some reports regarding the evaluation of chromatin integrity of fresh, chilled, and frozen-thawed dog sperm [12,20,21]; however, all of them have been performed using the SCSA. Few studies have been recently published using the SCDt to assess fresh and frozen-thawed epididymal or ejaculated dog sperm DNA integrity [22][23][24]. To our knowledge, no studies have been performed to evaluate canine chilledstored semen samples using the SCDt, a simple and costeffective method.…”