Animals

Research

Jump to: Review

10 pages, 901 KiB

Open AccessArticle

Hypo-Osmotic Swelling Test (HOST) for Feline Spermatozoa: The Simplified Procedure and the Aspect of Sperm Morphology

by Sylwia Prochowska, Wojciech Niżański and Alain Fontbonne

Animals 2022, 12(7), 903; https://doi.org/10.3390/ani12070903 - 31 Mar 2022

Cited by 6 | Viewed by 5729

Abstract

Hypo-osmotic swelling test (HOST) is used to assess the functional integrity of sperm plasma membranes in many species. The primary aim of this study was to test a simplified HOST procedure for the evaluation of feline semen. The second objective was to check [...] Read more.

Hypo-osmotic swelling test (HOST) is used to assess the functional integrity of sperm plasma membranes in many species. The primary aim of this study was to test a simplified HOST procedure for the evaluation of feline semen. The second objective was to check if sperm abnormalities can influence the results of this test. Urethral semen was collected from 19 male, domestic cats. In Exp. 1, HOST was performed in different media (50 mOsm/kg fructose or distilled water), temperature (37 °C or room temperature) and time (5 and 30 min). In Exp. 2, the potential effect of sperm abnormalities on HOST results was assessed by observing individual normal and abnormal spermatozoa microinjected into droplets of distilled water. The results showed no differences between the HOST results performed in different media, temperature and time. Viable abnormal spermatozoa were able to swell under hypo-osmotic conditions in the same manner as normal ones, except spermatozoa with distal droplets, which showed a higher frequency of ‘despiralization’. In conclusion, HOST can be reliably performed at 0 mOsm/kg for 5 min at room temperature, which may contribute to a wider use of this test under clinical environments. Viable abnormal spermatozoa are able to swell under hypo-osmotic conditions; therefore, their presence in the ejaculate would not bias the results of HOST when total coiling is calculated. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

15 pages, 20364 KiB

Open AccessArticle

Expression of Anti-Müllerian Hormone and Its Type 2 Receptor in the Ovary of Pregnant and Cyclic Domestic Cats

by Nilgün Gültiken, Murat Yarim, Selim Aslan, Hande Gürler, Gul Fatma Yarim, Müge Tuncay, Sinem İnal and Sabine Schäfer-Somi

Animals 2022, 12(7), 877; https://doi.org/10.3390/ani12070877 - 30 Mar 2022

Cited by 8 | Viewed by 2108

Abstract

To evaluate the expression of AMH and its receptor AMHRII, ovaries of 33 p cats were investigated by western blot and immunohistochemistry. After ovariohysterectomy, the cats were grouped according to pregnancy stages and ovarian/placental endocrine activity: group I (n = 3, 24–29 [...] Read more.

To evaluate the expression of AMH and its receptor AMHRII, ovaries of 33 p cats were investigated by western blot and immunohistochemistry. After ovariohysterectomy, the cats were grouped according to pregnancy stages and ovarian/placental endocrine activity: group I (n = 3, 24–29 days), II (n = 8, 32–40 days), III (n = 4, 41–46 days), IV (n = 6, 53–61 days) and according to cycle stages: V (n = 6, interestrus) and VI (n = 6, estrus). Serum progesterone- and AMH-concentration was measured. Follicle numbers did not differ between groups. The number of corpora lutea was higher in pregnant cats than in the non-pregnant cats. Serum AMH concentration was at maximum between day 30 and 50 of gestation, and was higher than in non-pregnant cats, then decreased towards term (p < 0.05). In the ovaries, AMH immunopositivity was observed in granulosa cells of secondary and antral follicles, and in interstitial cells of corpora lutea; highest percentage of immunopositive areas was detected in group III (p < 0.05). A positive correlation between the number of corpora lutea and the positive AMH signals in ovarian tissue was determined (r² = 0.832, p < 0.05); however, only during mid-gestation (group II). Expression of AMHRII was in close co-localization with AMH and strong in the interstitial cells surrounding follicles undergoing atresia. AMHRII expression did not differ between pregnant groups but was higher compared to estrus cats (p ˂ 0.05). We conclude that AMH and AMHRII expression in the feline ovary is comparable to other species. The high serum AMH concentration and ovarian AMHRII expression between day 30 and 50 of gestation are probably related to ovarian activity and follicular atresia. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

12 pages, 698 KiB

Open AccessArticle

Freezability of Dog Semen after Collection in Field Conditions and Cooled Transport

by Martina Colombo, Maria Giorgia Morselli, Giulia Franchi, Sabine Schäfer-Somi and Gaia Cecilia Luvoni

Animals 2022, 12(7), 816; https://doi.org/10.3390/ani12070816 - 23 Mar 2022

Cited by 3 | Viewed by 3495

Abstract

Dog semen freezing is gaining popularity, but it has to be performed in equipped facilities, which can be far from the place where the stud dog lives. The aim of this study was to evaluate whether freezing dog semen after 24 or 48 [...] Read more.

Dog semen freezing is gaining popularity, but it has to be performed in equipped facilities, which can be far from the place where the stud dog lives. The aim of this study was to evaluate whether freezing dog semen after 24 or 48 h of cooled transport to an equipped laboratory was possible when semen collection was performed in the field such as in local breeding kennels. Single ejaculates from different dogs (mixed breeds and ages) were collected. In Experiment I, 10 ejaculates were conventionally frozen using the Uppsala method or frozen after 24 or 48 h of storage in a Styrofoam transport box cooled by icepacks. In Experiment II, 10 ejaculates were used to assess the influence of two extenders (Uppsala chilling extender or freezing extender 1) used for semen dilution during the 24 or 48 h storage. Motility, morphology, membrane, and acrosome integrity were analyzed as well as spermatozoa zona-binding ability. No significant differences were observed among the frozen groups, regardless of freezing time (Experiment I) or extender (Experiment II). Motility at thawing, however, decreased in absolute value at 48 h. Freezing of freshly collected semen is the gold standard, but the results obtained in this study prompt the application of freezing after cooled transport for the long-term preservation of dog semen, especially if the transport can be organized in 24 h. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

18 pages, 2839 KiB

Open AccessArticle

Antigestagens Mediate the Expression of Decidualization Markers, Extracellular Matrix Factors and Connexin 43 in Decidualized Dog Uterine Stromal (DUS) Cells

by Ali Kazemian, Miguel Tavares Pereira, Bernd Hoffmann and Mariusz P. Kowalewski

Animals 2022, 12(7), 798; https://doi.org/10.3390/ani12070798 - 22 Mar 2022

Cited by 6 | Viewed by 2064

Abstract

Feto-maternal communication in the dog involves the differentiation of stromal cells into decidual cells. As the only placental cells expressing the nuclear progesterone (P4) receptor (PGR), decidual cells play crucial roles in the maintenance and termination of pregnancy. Accordingly, to investigate possible PGR-mediated [...] Read more.

Feto-maternal communication in the dog involves the differentiation of stromal cells into decidual cells. As the only placental cells expressing the nuclear progesterone (P4) receptor (PGR), decidual cells play crucial roles in the maintenance and termination of pregnancy. Accordingly, to investigate possible PGR-mediated mechanisms in canine decidual cells, in vitro decidualized dog uterine stromal (DUS) cells were treated with functional PGR-blockers, mifepristone and aglepristone. Effects on decidualization markers, epithelial and mesenchymal factors, and markers of cellular viability were assessed. Decidualization increased the expression of PTGES, PGR, IGF1, and PRLR, along with ECM1, COL4 and CX43, but downregulated IGF2. DUS cells retained their mesenchymal character, and the expression of COL4 indicated the mesenchymal-epithelial transformation. Antigestagen treatment decreased the availability of PTGES, PRLR, IGF1 and PGR. Furthermore, antigestagens decreased the mRNA and protein expression of CX43, and transcriptional levels of ECM1 and COL4. Additionally, antigestagens increased levels of activated-CASP3 (a proapoptotic factor), associated with lowered levels of PCNA (a proliferation marker). These data reveal important aspects of the functional involvement of PGR in canine decidual cells, regarding the expression of decidualization markers and acquisition of epithelial-like characteristics. Some of these mechanisms may be crucial for the maintenance and/or termination of canine pregnancy. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

13 pages, 1046 KiB

Open AccessArticle

Influence of Single Layer Centrifugation with Canicoll on Semen Freezability in Dogs

by Guillaume Domain, Hiba Ali Hassan, Eline Wydooghe, Osvaldo Bogado Pascottini, Anders Johannisson, Jane M. Morrell, Wojciech Niżański and Ann Van Soom

Animals 2022, 12(6), 714; https://doi.org/10.3390/ani12060714 - 11 Mar 2022

Cited by 5 | Viewed by 2170

Abstract

This study evaluated how semen selection by single layer centrifugation (SLC) with Canicoll affects semen freezability in dogs. A total of eighteen ejaculates, collected from dogs with optimal and suboptimal semen quality (optimal: normal morphology (NM) ≥ 80%, n = 9; suboptimal: NM [...] Read more.

This study evaluated how semen selection by single layer centrifugation (SLC) with Canicoll affects semen freezability in dogs. A total of eighteen ejaculates, collected from dogs with optimal and suboptimal semen quality (optimal: normal morphology (NM) ≥ 80%, n = 9; suboptimal: NM between 60 and 79%, n = 9), were divided into two aliquots and subjected to standard centrifugation or SLC before cryopreservation. Motility, NM, membrane integrity, mitochondrial membrane potential (MMP), and DNA integrity were improved in fresh samples after SLC, regardless of semen quality, but at the expense of some good quality spermatozoa. After thawing, NM and membrane integrity were improved in SLC-selected semen in both semen qualities. Interestingly, MMP was also higher but only in optimal quality semen. Still, spermatozoa from suboptimal quality semen did not survive freezing to the same extent as spermatozoa from optimal quality semen, even after selecting superior spermatozoa. Semen selection with Canicoll is, therefore, an effective technique to isolate a subpopulation of high-quality spermatozoa and obtain sperm samples of better quality after thawing, but is not sufficient to improve the intrinsic inferior freezability of suboptimal quality semen. So far, eighteen pups were born after insemination with SLC-selected frozen-thawed semen, proving that these selected spermatozoa remain fertile. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

9 pages, 1156 KiB

Open AccessArticle

Sperm Gone Smart: A Portable Device (iSperm^®) to Assess Semen Concentration and Motility in Dogs

by Guillaume Domain, Penelope Banchi, Hiba Ali Hassan, Anouk Eilers, Joke Lannoo, Eline Wydooghe, Wojciech Niżański and Ann Van Soom

Animals 2022, 12(5), 652; https://doi.org/10.3390/ani12050652 - 04 Mar 2022

Cited by 1 | Viewed by 2963

Abstract

The iSperm^® is a portable device for semen analysis. This study aimed to investigate its correlation with a conventional computer-assisted sperm analyzer (ISAS^®v1) for the assessment of semen concentration and kinematic parameters in dogs (n = 224). The intra-assay [...] Read more.

The iSperm^® is a portable device for semen analysis. This study aimed to investigate its correlation with a conventional computer-assisted sperm analyzer (ISAS^®v1) for the assessment of semen concentration and kinematic parameters in dogs (n = 224). The intra-assay variability of both devices and their ability to estimate semen concentration at a fixed value of 40 × 10⁶/mL were also investigated. Results showed that the intra-assay variability was lower for the ISAS^®v1 for all parameters compared to the iSperm^®. Hence, iSperm^® estimates were more variable in-between fields. Both the iSperm^® and the ISAS^®v1 were not reliable in estimating semen concentration (ISAS^®v1: median 30 × 10⁶/mL, interquartile range (IQR) 12, p < 0.01; iSperm^®: median 35.12 × 10⁶/mL, IQR 11.11, p < 0.01). Finally, positive correlations were found between both devices with stronger correlations obtained when four fields were analyzed by the iSperm^®. However, the low number of spermatozoa analyzed per field and the inability to avoid artifacts are downsides that currently limit the reliability of the iSperm^®. Therefore, the software of iSperm^® needs some improvement to make it a valid and practical alternative to automated computerized systems for the analysis of canine semen. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

8 pages, 543 KiB

Open AccessArticle

Serum Anti-Müllerian Hormone: A Potential Semen Quality Biomarker in Stud Dogs?

by Guillaume Domain, Justyna Buczkowska, Patrycja Kalak, Eline Wydooghe, Penelope Banchi, Osvaldo Bogado Pascottini, Wojciech Niżański and Ann Van Soom

Animals 2022, 12(3), 323; https://doi.org/10.3390/ani12030323 - 28 Jan 2022

Cited by 10 | Viewed by 2437

Abstract

Anti-Müllerian hormone (AMH) has been suggested to be involved in spermatogenesis. The aim of this study was to investigate the relationship between blood serum AMH concentration and semen quality in dogs. Moreover, this study sought to find the optimal cut-off point value of [...] Read more.

Anti-Müllerian hormone (AMH) has been suggested to be involved in spermatogenesis. The aim of this study was to investigate the relationship between blood serum AMH concentration and semen quality in dogs. Moreover, this study sought to find the optimal cut-off point value of serum AMH with the greatest sensitivity and specificity to predict semen quality. Forty-five clinically healthy dogs were included in the study and their age as well as the following semen parameters were determined and correlated to serum AMH concentration: total sperm output, normal morphology, plasma membrane integrity, total motility, progressive motility, and velocity parameters. Statistical analysis for correlations were performed using Spearman’s correlation coefficients. Moderate negative associations were found between serum AMH and semen total motility (r = −0.38, p = 0.01), progressive motility (r = −0.36, p = 0.01), and normal morphology (r = −0.36, p= 0.02). Based on these associations, an AMH concentration of 5.54 µg/L was found to be the optimal cut-off point value to obtain the greatest summation of sensitivity (86%) and specificity (63%) to predict semen quality. The serum AMH assay may therefore be a potential hormonal marker to predict which dogs would require further semen analysis. Future research is however needed to confirm these preliminary results. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

20 pages, 3380 KiB

Open AccessArticle

Utero-Placental Immune Milieu during Normal and Aglepristone-Induced Parturition in the Dog

by Miguel Tavares Pereira, Renata Nowaczyk, Selim Aslan, Serhan S. Ay and Mariusz P. Kowalewski

Animals 2021, 11(12), 3598; https://doi.org/10.3390/ani11123598 - 19 Dec 2021

Cited by 4 | Viewed by 2696

Abstract

Maternal immunotolerance is required for the maintenance of pregnancy, in sharp contrast with the uterine pro-inflammatory activity observed during parturition in several species. Correspondingly, in the dog, increased immune signaling at term has been suggested, but a deeper understanding of the uterine immune [...] Read more.

Maternal immunotolerance is required for the maintenance of pregnancy, in sharp contrast with the uterine pro-inflammatory activity observed during parturition in several species. Correspondingly, in the dog, increased immune signaling at term has been suggested, but a deeper understanding of the uterine immune milieu is still missing. Thus, the availability of 30 immune-related factors was assessed in utero-placental samples collected during post-implantation (days 18–25 of pregnancy) and mid-gestation (days 35–40) stages, and at the time of prepartum luteolysis. Gene expression and/or protein localization studies were employed. Samples collected from antigestagen (aglepristone)-treated dogs were further analyzed. Progression of pregnancy was associated with the downregulation of IL1β and upregulation of IL10 (p < 0.05) at mid-gestation. When compared with mid-gestation, a higher availability of several factors was observed at term (e.g., CD206, CD4, TLR4). However, in contrast with natural parturition, MHCII, CD25, CCR7, TNFα, IDO1 and AIF1 were upregulated after aglepristone treatment (p < 0.05), but not TNFR1 or CCL13 (p > 0.05). Altogether, these results show an increased immune activity during canine parturition, involving, i.a., M2 macrophages, Treg and Th cells, with strong support for progesterone-mediated immunomodulation. Furthermore, differences between term and induced parturition/abortion could relate to differences in placental maturation towards parturition and/or functional traits of antigestagens. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

5 pages, 530 KiB

Open AccessCommunication

Application of the FISH Technique to Visualize Sex Chromosomes in Domestic Cat Spermatozoa

by Barbara Kij-Mitka, Halina Cernohorska, Svatava Kubickova, Sylwia Prochowska, Wojciech Niżański, Joanna Kochan and Monika Bugno-Poniewierska

Animals 2021, 11(7), 2106; https://doi.org/10.3390/ani11072106 - 15 Jul 2021

Cited by 1 | Viewed by 3964

Abstract

Fluorescence in situ hybridization is a molecular cytogenetics technique that enables the visualization of chromosomes in cells via fluorescently labeled molecular probes specific to selected chromosomes. Despite difficulties in carrying out the FISH technique on sperm, related to the need for proper nuclear [...] Read more.

Fluorescence in situ hybridization is a molecular cytogenetics technique that enables the visualization of chromosomes in cells via fluorescently labeled molecular probes specific to selected chromosomes. Despite difficulties in carrying out the FISH technique on sperm, related to the need for proper nuclear chromatin decondensation, this technique has already been used to visualize chromosomes in human, mouse, cattle, swine, horse, and dog spermatozoa. Until now, FISH has not been performed on domestic cat sperm; therefore, the aim of this study was to visualize sex chromosomes in domestic cat sperm. The results showed the presence of X and Y chromosomes in feline spermatozoa. The procedure used for sperm decondensation and fluorescence in situ hybridization was adequate to visualize chromosomes in domestic cat spermatozoa and, in the future, it may be used to determine the degree of chromosomal abnormalities in these gametes. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

Review

Jump to: Research

12 pages, 721 KiB

Open AccessReview

Canine Spermatozoa—Predictability of Cryotolerance

by Sabine Schäfer-Somi, Martina Colombo and Gaia Cecilia Luvoni

Animals 2022, 12(6), 733; https://doi.org/10.3390/ani12060733 - 15 Mar 2022

Cited by 2 | Viewed by 2493

Abstract

Markers of freezability allow the selection of ejaculates of good freezability. So far, most investigations were conducted in boars, bulls, rams and horses, with high economic interests triggering the efforts. The progress in dogs is comparably slow. A critical evaluation of the methods [...] Read more.

Markers of freezability allow the selection of ejaculates of good freezability. So far, most investigations were conducted in boars, bulls, rams and horses, with high economic interests triggering the efforts. The progress in dogs is comparably slow. A critical evaluation of the methods requires consideration of practicability, with most labs not even possessing a computer assisted sperm analyser (CASA); furthermore, small canine ejaculates mostly do not allow the use of large semen volumes. In dogs, modern markers of freezability no longer assess single membrane constituents or seminal plasma components but comprise tests of cell functionality and adaptability, energy metabolism, cluster analyses of kinetic and morphometric parameters, as well as DNA intactness. Identification of the most efficient combination of tests seems useful. At present, examination by CASA combined with cluster analysis of kinetic subgroups, JC-1 staining and COMET assay or staining with toluidine blue seem most appropriate; however, cell volumetry and other functional tests deserve better attention. A better understanding of spermatozoa energy metabolism might reveal new markers. This review focuses on the requirements and markers of freezability of canine semen, highlighting potential future candidates. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

10 pages, 32522 KiB

Open AccessReview

Canine and Feline Testicular Preservation

by Lúcia Daniel Machado da Silva

Animals 2022, 12(1), 124; https://doi.org/10.3390/ani12010124 - 05 Jan 2022

Cited by 2 | Viewed by 2504

Abstract

The increased interest in breeding dogs and cats and their use as models for other canids and felids demand research to improve reproductive techniques. Among them, testicular cryopreservation stands out. Testicular cryopreservation enables the maintenance of reproductive capacity and allows the establishment of [...] Read more.

The increased interest in breeding dogs and cats and their use as models for other canids and felids demand research to improve reproductive techniques. Among them, testicular cryopreservation stands out. Testicular cryopreservation enables the maintenance of reproductive capacity and allows the establishment of germplasm banks for several species of commercial value or at risk of extinction. Furthermore, it enables the transport of genetic material among different regions. It is noteworthy that this biotechnology represents the only possibility of preserving the fertility of prepubertal animals that have died, so it has great importance in the propagation of the genetic material of animals. The spermatogonia present in the testes can be cultivated in vitro and the sperm obtained can be used in artificial reproduction programs. Although advances have been achieved with the use of testicular fragments to obtain viable and functional germ cells, the establishment of protocols that can be used in clinical routine have not been concluded yet. The testicular cryopreservation process can be carried out through techniques such as slow freezing, fast freezing and vitrification. However, the protocols used for the canine and feline species are still in the experimental phase. Given the importance of the topic, the aim of this review is to draw a profile of the subject approaching the main works on testicular cryopreservation in dogs and cats. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

10 pages, 876 KiB

Open AccessReview

Canine and Feline Epididymal Semen—A Plentiful Source of Gametes

by Hiba Ali Hassan, Guillaume Domain, Gaia Cecilia Luvoni, Rana Chaaya, Ann Van Soom and Eline Wydooghe

Animals 2021, 11(10), 2961; https://doi.org/10.3390/ani11102961 - 14 Oct 2021

Cited by 10 | Viewed by 6482

Abstract

Canine and feline epididymal semen provide an additional source of gametes to preserve the genetics of valuable breeding dogs and tomcats, especially for those that fail to ejaculate, need castration as a therapy or die unexpectedly. Moreover, since it is quite common to [...] Read more.

Canine and feline epididymal semen provide an additional source of gametes to preserve the genetics of valuable breeding dogs and tomcats, especially for those that fail to ejaculate, need castration as a therapy or die unexpectedly. Moreover, since it is quite common to perform castration of non-breeding dogs and cats, the development of a gene bank of epididymal semen collected after castration would greatly contribute to increase the genetic diversity in dogs and cats. Collection and cryopreservation of epididymal semen necessitates a full understanding of the function of the epididymis and of the characteristics of epididymal spermatozoa as opposed to ejaculated semen. During collection of epididymal semen, specific factors may have a negative effect on epididymal semen quality and freezability. Accordingly, the elimination of these triggers could enhance epididymal semen freezability and consequently positively influence post-thaw semen quality and outcome for different ARTs. Full article

(This article belongs to the Special Issue Biotechnology and Reproduction in Companion Animals)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Biotechnology and Reproduction in Companion Animals

Share This Special Issue

Special Issue Editor

Special Issue Information

Keywords

Published Papers (12 papers)

Research

Review

Further Information

Guidelines

MDPI Initiatives

Follow MDPI