Analysis of morphological disorders and ploidy in domestic cat blastocysts.

This study describes, for the first time, the relationship between morphology and ploidy in domestic cat embryos. Blastocyst morphology and quality were assessed using time-lapse recordings, while ploidy was analyzed using fluorescence in situ hybridization. Out of 54 blastocysts, clear fluorescence signals for all the molecular probes used were observed in 24 (44.4%) blastocysts, while in another 14 (25.9%) blastocysts, fluorescence signals only allowed for sex assessment. No clear signals were observed in the remaining 16 blastocysts (29.7%). Of the 24 blastocysts with clear signals, normal ploidy was detected in 10 (41.4%), 7 (29.2%) were diagnosed as haploid, and the remaining 7 blastocysts (29.2%) were mosaics. Additionally, results showed the distribution of diploid, haploid, and mosaic blastocysts in relation to the occurrence of morphological disorders and to embryo quality. The presence of abnormal embryo morphology and karyotype disorders may affect further development and the pregnancy rate. Due to the comparable proportion of good and poor quality blastocysts with disturbed ploidy, it is important to implement new methods of embryo assessment, especially when techniques used in humans, such as pronuclear observation, cannot be used.

Transscrotal stimulation of the testes and epididymides improves urethral sperm collection in domestic cats.

Urethral catheterisation after medetomidine administration is the method of choice for semen collection in cats, but it yields variable results. This study tested whether scrotal manual stimulation can improve urethral sperm collection in domestic cats. The study was performed on 20 male cats, from which two urethral semen samples were collected, one before and one after 2min of transscrotal finger massage of the testes and epididymides. Both sperm samples were assessed for total sperm count and motility using computer-aided sperm analysis, viability and morphology (eosin-nigrosin staining). The transscrotal manual stimulation allowed a significantly higher number of spermatozoa to be obtained (P=0.0015). Viability was similar before and after the stimulation (median 92% and 90.5%), whereas the number of motile (median 60% and 70%) and morphologically normal (median 17% and 30.5%) spermatozoa was higher in the second sample (P=0.03 and P=0.002 respectively), which confirms that transscrotal massage induced the expulsion of a fresh pool of spermatozoa into the urethra. Transscrotal stimulation of the testes and epididymides significantly improves urethral semen collection in domestic cats and can be easily introduced into clinical practice.

The Viability of Serval (Leptailurus serval) and Pallas Cat (Felis manul) Oocytes after Cryopreservation Using the Rapid-I Method.

BACKGROUND: Vitrification by Rapid-I method could be essential for felid rescue programs to protect wild felid in the future. OBJECTIVE: This study was aimed at adapting the Rapid I method and evaluating the viability of serval and Pallas cat oocytes compared to oocytes of the domestic cat. MATERIALS AND METHODS: Oocytes after collection and in vitro maturation were vitrified using Cryotech medium (Cryotech, Japan) and a Rapid-I device (Vitrolife, Sweden). To evaluate viability, oocytes after warming were stained with fluorescein diacetate and ethidium bromide. RESULTS: Survival rate in the control group (domestic cat) was 75 %. In the experimental group, 70% (serval) and 60% (pallas cat) viable oocytes were found. CONCLUSION: The Rapid-I method can be applied successfully for the vitrification of wild felid oocytes.

In vitro fertilizing potential of urethral and epididymal spermatozoa collected from domestic cats (Felis catus).

The aim of this study was to provide a comparative analysis of in vitro fertilizing potential of frozen-thawed urethral and epididymal feline spermatozoa. Both types of semen were collected from 7 cats and cryopreserved in liquid nitrogen. To perform in vitro fertilization, both urethral and epididymal samples from the same individual were thawed and spermatozoa were co-incubated with in vitro matured cat oocytes. Obtained embryos were cultured in vitro for 7 days in a commercial medium. Cleavage rate, morula rate and blastocyst rate were calculated. Experiment was run in 10 replicates. The examined parameters showed no significant differences between urethral and epididymal spermatozoa (p>0.05). Cleavage rate and embryo's development were highly variable between replicates, even for the different sperm samples collected from one individual. There was no significant correlation between fertilizing capacity of two types of spermatozoa collected from the same male. In this study we confirmed that cryopreserved urethral spermatozoa have equally good fertilizing potential as epididymal ones, and both can be successfully used for in vitro fertilization in cats with the use of commercial medium.

Evaluation of spermatozoal function-useful tools or just science.

Conventional microscopic semen analysis does not provide precise information on the fertilizing potential of a male. The traditional basis for semen evaluation is that male fertility is dependent on production of a "proper" concentration/number of motile, morphologically normal spermatozoa in excess to achieve conception. Many independent studies, especially in human medicine, have demonstrated that the absolute number of spermatozoa does not accurately determine fertility, but their functional competence. Many functional tests of spermatozoa are developed over the last decades. Computer-assisted sperm analysis (CASA) and flow cytometry have become the gold standard for semen assessment in specialized andrology laboratories. Other functional assays, such as gamete interaction tests, provide additional information regarding the real fertilizing potential of sperm cells. From this point of view, such tests are valuable diagnostic tools in fertility disorders and may be helpful to make a decision which method of treatment to use: pharmacological therapy, intrauterine insemination, introduction of classic IVF, ICSI or exclusion from a breeding programme. The most useful gamete interaction tests include induced acrosome reaction, zona pellucida binding assay, oocyte penetration assay and hyaluronan binding assay. In recent years, andrology has entered into a new era of sophisticated OMICS methods. Genomics, epigenomics, transcriptomics and proteomics brought high hopes for rapid progress in clinical diagnostics.