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1.
Pol J Vet Sci ; 26(3): 497-509, 2023 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-37727970

RESUMEN

The main cause of sperm chromatin damage is oxidative stress related to embryo development failure and adult infertility in mammals and also avian. Oxidative stress results in lipid peroxidation (LPO) causing cell damage. Lipid peroxidation is the oxidation of polyunsaturated fatty acids (PUFAs) in biological systems and causes changes in the physical structure and characteristics of the cell membrane. Due to the high amounts of PUFAs in the avian sperm membrane, its sperm seem susceptible to pe-roxidative damage and is a substantial factor in the fertilization capacity of sperm. The most commonly used methods for measuring LPO or its by-products, such as malondialdehyde (MDA) and 4-hydroksy-2-nonenal (4-HNE), in bird semen are based on the colorimetric method TBARS (thiobarbituric acid reactive substances) and on the use of a fluorescence probe (CC 11-BODIPY 581/591) as a marker to evaluate membrane lipid peroxidation. This review aims first to introduce LPO in avian semen and its effects on avian sperm and second to summarize the commonly applied methods of evaluating LPO and its damage in fresh and stored avian semen.


Asunto(s)
Líquidos Corporales , Semen , Masculino , Animales , Peroxidación de Lípido , Espermatozoides , Aves , Mamíferos
2.
Theriogenology ; 186: 114-121, 2022 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-35462315

RESUMEN

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.


Asunto(s)
Blastocisto , Ploidias , Animales , Gatos , Femenino , Hibridación Fluorescente in Situ/veterinaria , Embarazo , Índice de Embarazo
3.
Anim Genet ; 50(6): 705-711, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31476086

RESUMEN

The genetic background of disorders of sex development (DSD) in dogs with a normal male sex chromosome set (78,XY) is poorly described. In this study, we present for the first time, an analysis of six genes of the testosterone pathway, encoding enzymes (CYP17A1, HSD3B2, HSD17B3, SRD5A2) and transcription factors (NR5A1, AR). The entire coding sequence and flanking regions of the introns, 5'-UTR and 3'-UTR were analyzed in five DSD dogs (78,XY, SRY-positive) with ambiguous external genitalia and in 15 control dogs. A homozygous deletion of 2 bp in exon 2 of HSD17B3 (hydroxysteroid 17-beta dehydrogenase 3) was found in a Dachshund dog with enlarged clitoris, vulva and abdominal gonads and decreased serum testosterone level. In silico analysis revealed that this deleterious variant causes truncation of the encoded polypeptide (from 306 to 65 amino acids) and deprivation of the active site of the encoded enzyme. Genotyping of 23 control Dachshund dogs showed a normal homozygous genotype. Thus, we assumed that the 2-bp deletion is the causative variant. Moreover, 24 SNPs (four in CYP17A1, three in HSD3B2, six in HSD17B3, five in SRD5A2, one in AR and five in NR5A1), two intronic indels (one in HSD3B2 and one in SRD5A2) and two microsatellite polymorphisms in exon 1 of AR were found. Six SNPs appeared to be novel. No association with DSD phenotype was observed. Identification of the first case of DSD in domestic animals caused by a deleterious variant of a gene involved in testosterone synthesis showed that these genes are important candidates in such studies.


Asunto(s)
Trastornos del Desarrollo Sexual/veterinaria , Enfermedades de los Perros/genética , Testosterona/biosíntesis , 17-Hidroxiesteroide Deshidrogenasas/genética , Animales , Codón de Terminación , Trastornos del Desarrollo Sexual/genética , Trastornos del Desarrollo Sexual/patología , Perros , Femenino , Eliminación de Gen , Genitales/patología , Masculino , Testosterona/sangre
4.
Pol J Vet Sci ; 22(2): 305-312, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31269335

RESUMEN

AIM: The aim of this study was to analyze the effect of bovine follicular fluid on the survival, morphology and kinetic parameters of bovine thawed spermatozoa under laboratory conditions. MATERIALS AND METHODS: The semen from 5 bulls of proven fertility was incubated in follicular and physiological fluid for 8 hours. During this time assessment using the CASA system was performed. At the beginning and the end of incubation process evaluation by flow cytometry was conducted. RESULTS: The results of the sperm motility assessment showed a significant decrease in the analyzed parameters both in the follicular and physiological fluid. A significant reduction in all parameters characterizing movement properties in the semen incubated in the follicular fluid was found. In the physiological fluid, a similar trend was demonstrated only for the following properties: VAP, VSL, VCL, ALH, BCF. A significant difference was found for both fluids in: VCL (p=0.026), ALH (p=0.038) and LIN (p⟨0.001) at the beginning of incubation. The results of the plasma membrane integrity assessment showed a statistically significant increase in the percentage of dying sperm at the 8th hour of the incubation in the follicular fluid. In the case of semen incubation in physiological fluid, a statistically significant decrease in the percentage of live non-damaged cells was found with a simultaneous increase in the subpopulation of undamaged dead cells. CONCLUSIONS: Follicular fluid rapidly accelerates the capacitation process. The results of flow cytometry support the hypothesis concerning the ability of follicular fluid to prolong sperm survival.


Asunto(s)
Bovinos , Citometría de Flujo/veterinaria , Líquido Folicular , Procesamiento de Imagen Asistido por Computador/instrumentación , Análisis de Semen/veterinaria , Espermatozoides/efectos de los fármacos , Animales , Criopreservación/veterinaria , Femenino , Procesamiento de Imagen Asistido por Computador/métodos , Masculino , Preservación de Semen/veterinaria
5.
Cryo Letters ; 40(4): 226-230, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31278403

RESUMEN

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.


Asunto(s)
Supervivencia Celular , Criopreservación/veterinaria , Felidae , Felis , Oocitos/citología , Animales , Criopreservación/métodos , Crioprotectores , Femenino , Vitrificación
6.
Reprod Domest Anim ; 53(3): 818-821, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29542185

RESUMEN

Wisent, or European bison (Bison bonasus), is listed as "vulnerable" on the IUCN Red List of Threatened Species and is therefore protected by international law. For the first time, a Wisent embryo has been obtained in vitro. This procedure creates a new opportunity to protect and increase Wisent reproductive potential and thereby opens new possibilities for the establishment of a controlled and broad reserve of the gene pool.


Asunto(s)
Bison/fisiología , Técnicas de Cultivo de Embriones/veterinaria , Fertilización In Vitro/veterinaria , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Animales , Blastocisto , Chlorocebus aethiops , Técnicas de Cocultivo , Técnicas de Cultivo de Embriones/métodos , Especies en Peligro de Extinción , Femenino , Fertilización In Vitro/métodos , Técnicas de Maduración In Vitro de los Oocitos/métodos , Masculino , Células Vero
7.
Theriogenology ; 102: 59-66, 2017 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-28750295

RESUMEN

This study investigated the effect of exposure to zearalenone (ZEN) and its metabolites on the characteristics of epididymal spermatozoa, testicular and epididymal biometry and histology, and the concentration of testosterone in blood plasma in male wild boars. The study was performed during more than one year on 18 clinically healthy male wild boars with initial and final body weight, of 39 ± 4 kg and 59 ± 3 kg, respectively. The animals were divided into two experimental groups (group I and group II) and one control group (group C) comprising 6 boars per group. Group I animals were administered per os pure zearalenone (ZEN) at 150 µg/kg BW for 7 consecutive days every two months, while group II animals received a dose of 50 µg/kg BW/day via feed that was naturally contaminated with ZEN. These male wild boars were exposed to ZEN over a period of 1 year. Control animals were fed a placebo. Testicles with epididymides of the boars were collected on the last day of the experiment within 3 min after slaughter. Blood samples were collected from each of the male wild boars. Testes and epididymides were measured and sampled for histological examination. Epididymides were dissected and epididymal spermatozoa were harvested. The spermatozoa were diluted with swine-specific BTS extender and stored at 17°C for 144 h. Sperm motility was analyzed with CASA, and other parameters including viability, acrosome integrity, DNA fragmentation index, lipid peroxidation and apoptosis were assessed with flow cytometry. In these wild boars, per os exposure to natural sources of ZEN or a combination of ZEN and its metabolites changed the testicular interstitium and led to modification of some epididymal sperm parameters. The interstitial glands in testes of experimental group I were markedly reduced and hyperemic with evident blood stasis in small capillaries. Also in group I were single degenerating seminiferous tubules. In both groups I and II, immediately after dilution of spermatozoa with BTS remarkable decreases in motility rate as well as in progressive motility and the subpopulation of cells with rapid movement were noted compared with the control group (P < 0.05). But unexpectedly, after 24 h incubation of boar semen in the BTS diluent, these sperm properties improved and were not significantly different from the control group. Thus, exposure to ZEN has no lasting but only a temporary, reversible effect on wild boar sperm motility. There was no influence of exposure to ZEN and its metabolites on the integrity of membranes, intensity of lipid peroxidation and apoptosis or on sperm chromatin structure. This study is the first using these direct measures of sperm motility and integrity to show a redundant adverse effect of ZEN exposure on wild boar sperm characteristics. There were no effects of exposure to ZEN and its metabolites on body weight, testicular and epididymal biometry, gonadosomatic index and the concentration of testosterone in blood plasma in the male wild boars.


Asunto(s)
Espermatozoides/efectos de los fármacos , Sus scrofa/fisiología , Testosterona/sangre , Zearalenona/toxicidad , Animales , Epidídimo/fisiología , Estrógenos no Esteroides/toxicidad , Peroxidación de Lípido/efectos de los fármacos , Masculino , Sus scrofa/sangre , Testículo
8.
J Dairy Sci ; 100(8): 6707-6719, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28551182

RESUMEN

Despite recent advances in bull epididymal fluid proteome research, significant numbers of proteins secreted to epididymal lumen remain unidentified. The objective of this study was to expand the number of identified cauda epididymal fluid proteins in bulls and to contextualize them in a broader view of their mutual interactions and involvement in biological processes and pathways, to fully elucidate the ways in which epididymal fluid proteins are involved in storage and maturation of spermatozoa in epididymis. We collected postmortem cauda epididymal fluid from 6 mature Holstein Friesian bulls. We performed the identification of proteins using 2-dimensional electrophoresis coupled with MALDI mass spectrometry. Analysis of functionality and pathway involvement of identified proteins was performed using Ingenuity Pathway Analysis software. We identified a total of 189 epididymal fluid proteins, out of which 100 were newly identified in bull epididymal fluid. We have combined our data with 2 previously performed bull epididymal fluid proteome identifications, yielding 280 proteins total, and analyzed it. The main canonical pathways involving epididymal proteins were glycolysis, gluconeogenesis, protein ubiquitination pathway, nuclear factor-erythroid 2-related factor 2-mediated oxidative stress response, and farnesoid X receptor/retinoid X receptor activation. The main biological functions potentially performed by epididymal fluid proteins included carbohydrate metabolism, cellular growth and proliferation, cell death and survival, and small molecule biochemistry. Overall, our results have pointed out multiple novel pathways in bull epididymal fluid that might take part in various aspects of maturation and protection processes of epididymal spermatozoa.


Asunto(s)
Proteínas de Plasma Seminal/aislamiento & purificación , Proteínas de Plasma Seminal/fisiología , Animales , Líquidos Corporales , Bovinos , Epidídimo , Masculino , Proteoma/metabolismo , Vesículas Seminales , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Espermatozoides
9.
Pol J Vet Sci ; 20(1): 19-24, 2017 Mar 28.
Artículo en Inglés | MEDLINE | ID: mdl-28525327

RESUMEN

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.


Asunto(s)
Gatos/fisiología , Criopreservación/veterinaria , Epidídimo/fisiología , Fertilización In Vitro/veterinaria , Espermatozoides/fisiología , Uretra/fisiología , Animales , Masculino , Análisis de Semen
10.
Pol J Vet Sci ; 20(1): 123-131, 2017 Mar 28.
Artículo en Inglés | MEDLINE | ID: mdl-28525341

RESUMEN

Disorders of sex development (DSD) are rare in cats. They can be caused by chromosomal aberrations, gene mutations or other undefined factors. The aim of the present study was to compare the histological structure and immunohistochemical reactivity of testes in cats with DSD and in healthy cats. The research material consisted of the gonads of four cats - phenotypic males with an incorrect structure of the reproductive system. The control group consisted of the testes of four healthy cats - routinely castrated phenotypical males. The material was fixed with formalin and embedded in paraffin; the sections were stained with hematoxylin and eosin. The immunohistochemical investigation were performed using monoclonal and polyclonal antibodies directed against desmin, vimentin, actin of smooth muscles, S100 protein and MCM3 protein. The results obtained allow concluding that the testes of cats with DSD differed in certain respects, mainly in the number of blood vessels, from the normal testes. Moreover, the results of immunohistochemical examination indicate that in the testes of cats with DSD the number of supporting cells is lower, the amount of interstitial cells is comparable and spermatogenesis is correct es compared to those determined in the control gonads. The number of blood vessels in cats with DSD is reduced by about 30%. It confirms the recommendations for castration of these animals in order to eliminate the potential inheritance of sex development disorders.


Asunto(s)
Enfermedades de los Gatos/patología , Trastornos del Desarrollo Sexual/veterinaria , Testículo/patología , Animales , Estudios de Casos y Controles , Gatos , Trastornos del Desarrollo Sexual/patología , Cariotipo , Masculino
11.
Reprod Domest Anim ; 52(5): 806-813, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28464344

RESUMEN

CONTENTS: The objective of this study was to evaluate mitochondria in immature and in vitro-matured domestic cat oocytes and to assess for the first time the effect of vitrification on mitochondrial traits. Mitochondrial distribution and aggregation were assessed using confocal microscopy after staining with the fluorescent dye-MitoTracker® Red CMXRos. Only cells at the germinal vesicle and the metaphase II stages of nuclear development, representing immature and mature oocytes, respectively, were included in our study. Our study shows that 80% of immature and 100% of mature oocytes exhibit a peripheral pattern of mitochondria distribution, indicating that, in contrast to the situation in other species, the mitochondria of cat oocytes are not dispersed throughout the cell after in vitro maturation but instead maintain a strong affinity for the oocyte periphery near the membrane. However, a loss of aggregation was observed during in vitro maturation-78% of immature oocytes showed homogeneous granulation versus only 18% of mature oocytes (p < .001). The increased intensity of MitoTracker® Red CMXRos staining after in vitro maturation (p < .05) may be tentatively attributed to an increase in mitochondrial activity but could likewise reflect a concomitant appearance of sulphhydryl groups in cytoplasm (known to be targeted by the dye). Mitochondrial distribution did not change upon vitrification; however, dye intensity decreased (p < .05) and mitochondrial aggregation was intensified in both immature and mature vitrified cat oocytes.


Asunto(s)
Gatos , Técnicas de Maduración In Vitro de los Oocitos , Mitocondrias/fisiología , Oocitos/crecimiento & desarrollo , Vitrificación , Animales , Criopreservación/veterinaria , Femenino , Oocitos/citología
12.
J Dairy Sci ; 100(3): 2282-2298, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28041731

RESUMEN

The existing knowledge on the bull seminal vesicle proteome, a major seminal plasma constituent, and its relationship with seminal plasma is limited. This knowledge is prerequisite for a better understanding of seminal plasma variability, which is linked to semen quality. The objective of this study was to characterize the proteomes of seminal vesicle fluid and seminal plasma and to compare them to better understand the origin of seminal plasma proteins. We collected ejaculates and seminal vesicle fluid postmortem from 6 mature Holstein Friesian bulls. We performed the analysis and identification of proteins using 2-dimensional electrophoresis coupled with matrix-assisted laser desorption/ionization mass spectrometry. We identified 105 proteins in bull seminal vesicle fluid and 88 proteins in seminal plasma. For both seminal vesicles and seminal plasma proteins described in our study, top biological functions were cellular movement, cell death and survival, and cellular growth and proliferation. Additionally, seminal vesicle fluid proteins were involved in protein degradation and synthesis. Seminal plasma proteins were also involved in cellular assembly and organization and cell-to-cell signaling and interactions. Proteins of both fluids were involved in the following canonical pathways: glycolysis, gluconeogenesis, liver X receptor/farnesoid X receptor, and farnesoid X receptor/retinoid X receptor activation. Additionally, seminal vesicle fluid proteins appeared to be involved in oxidative stress response mediated by nuclear factor E2-related factor 2. Our results described the bull seminal vesicle fluid proteome for the first time and allowed for significant expansion of the current knowledge on the bull seminal plasma proteome. Moreover, analysis indicated that both bull seminal vesicle fluid and seminal plasma proteomes contained interconnected protein groups related to protective functions, glycolysis, and the morphology and physiology of the spermatozoa. These proteins and their interactions could be targeted in future research.


Asunto(s)
Proteoma/metabolismo , Proteínas de Plasma Seminal , Animales , Bovinos , Masculino , Semen , Análisis de Semen , Vesículas Seminales , Espermatozoides
13.
Reprod Domest Anim ; 52 Suppl 2: 281-287, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-27859772

RESUMEN

Embryo vitrification is a modern technique for cryopreservation in assisted reproductive programs. From all the embryos, blastocysts are the most challenging during cryostorage due to their size, multicellular structure and the presence of blastocoelic fluid. The aim of this study was to evaluate the suitability for vitrification of various developmental stages of feline embryos and the influence of the artificial shrinkage (AS) of expanded blastocyst on post-vitrification survival rates. The AS procedure is the manual puncture of the trophectoderm allowing for the reduction of blastocoelic fluid prior to vitrification and thus preventing the ice crystal formation. The vitrified embryos were divided into groups of 2-cell, 4- to 8-cell, >8-cell, morulae, compacted and expanded blastocyst, based on morphological assessment and vitrified in groups of 1-3 embryos per Cryotop. The post-warming survival was similar regardless the embryo developmental stage prior vitrification; however, development to blastocysts was only noted in 4- to 8-cell and >8-cell vitrified embryos (13% and 27%, respectively). Following warming, the significantly more viable blastocysts were noted in vitrified compacted versus expanded blastocyst and in expanded blastocyst subjected to AS procedure versus expanded blastocyst without AS (total survival: 58.3% vs. 33.3% and 64.3% vs. 38.5%, respectively; re-expansion rate within 2 hr post-warming: 41.7 vs. 6.7% and 50% vs. 7.7%, respectively). One-fifth of vitrified expanded blastocyst showed morphological damage immediately after warming procedure, whereas no visible damage was noted in compacted blastocyst and artificially collapsed expanded ones. The obtained results suggest that the most suitable for vitrification are feline embryos containing four to 16 blastomeres and compacted blastocyst. In addition, the reduction of blastocoel cavity in expanding blastocyst by artificial collapse improved the blastocyst vitrification outcome.


Asunto(s)
Blastocisto/fisiología , Gatos/embriología , Criopreservación/veterinaria , Desarrollo Embrionario , Animales , Blastocisto/citología , Líquidos Corporales , Criopreservación/métodos , Femenino , Calor
14.
Reprod Domest Anim ; 51 Suppl 1: 37-45, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27670939

RESUMEN

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.

15.
Reprod Domest Anim ; 51(3): 378-85, 2016 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-27037507

RESUMEN

Predicting male fertility on non-invasive sperm traits is of big importance to human and animal reproduction strategies. Combining the wide range of parameters monitored by computer-assisted sperm analysis (CASA) with some molecular traits (e.g. mtDNA content) may help to identify markers of the male fertility. The aim of this study was to characterize variation in the mtDNA copy number in equine sperm and to investigate whether mtDNA content is correlated with quality traits of stallion spermatozoa and the age of the male. Ejaculates collected from 53 fertile stallions were divided into four age groups (3-5, 6-10, 11-14 and >15 years) and were subjected to a complex investigation including conventional analysis, CASA, flow cytometry and mtDNA content (real-time PCR). The mean (±SD) number of mtDNA copies equalled 14 ± 9 and varied from 3 to 64. Considering the great number of sperm parameters monitored in this study, only few of them were correlated with the mtDNA content: ejaculate volume (a positive correlation), the amplitude of lateral head displacement (ALH; a negative correlation) and the high mitochondrial activity index (a negative correlation). The stallion age was not correlated with the mtDNA copy number. This study provides the first set of data on mtDNA content in equine sperm and confirms phenomena previously described for humans and dog on associations between sperm mtDNA content and selected motility parameters monitored by the CASA. Basing our study on spermatozoa from fertile stallions could however limit the extent of detected associations.


Asunto(s)
ADN Mitocondrial/análisis , Fertilidad/genética , Caballos/genética , Espermatozoides/química , Factores de Edad , Animales , Perros , Marcadores Genéticos , Humanos , Masculino , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Análisis de Semen/veterinaria , Motilidad Espermática/genética
16.
Reprod Domest Anim ; 51(3): 339-45, 2016 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-26991408

RESUMEN

The aim of this study was to evaluate the developmental kinetics of cats' blastocysts in connection with their morphology and blastomeres allocation to trophoblast or embryoblast cells. We examined gross blastocyst morphology and the total number of blastomeres together with its allocation to inner cell mass (ICM) or trophectoderm (TE) cells in pre-implantation feline embryos obtained from 6th to 9th day of in vitro culture. From all the investigated embryos, 61.8% developed to early blastocyst, 37.4% to full and 7.6% to hatching blastocyst stage. The total cell number (TCN) varied form 58 cells in early day 6 to 245 in hatching day 8 blastocyst, with the mean 84.9 cells in early, 156.7 in full, and 204.4 in hatching ones. Day 8 blastocyst had the highest number of total cells, together with the highest mean number of ICM regardless of its morphological assessment. Early blastocyst (apart from day 6) had the highest number of arrested cells, while dead cells were the highest in full day 9 blastocyst. More data about the relationship between blastocyst development and morphology would facilitate the selection of optimal blastocysts for further procedures.


Asunto(s)
Blastocisto/citología , Gatos/embriología , Animales , Blastómeros/citología , Recuento de Células , Técnicas de Cultivo de Embriones/veterinaria , Desarrollo Embrionario , Fertilización In Vitro/veterinaria , Edad Gestacional , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Trofoblastos/citología
17.
Anim Genet ; 47(2): 245-9, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26692319

RESUMEN

Obesity is an emerging health problem in purebred dogs. Due to their crucial role in energy homeostasis control, genes encoding adipokines are considered candidate genes, and their variants may be associated with predisposition to obesity. Searching for polymorphism was carried out in three adipokine genes (TNF, RETN and IL6). The study was performed on 260 dogs, including lean (n = 109), overweight (n = 88) and obese (n = 63) dogs. The largest cohort was represented by Labrador Retrievers (n = 136). Altogether, 24 novel polymorphisms were identified: 12 in TNF (including one missense SNP), eight in RETN (including one missense SNP) and four in IL6. Distributions of five common SNPs (two in TNF, two in RETN and one in IL6) were further analyzed with regard to body condition score. Two SNPs in the non-coding parts of TNF (c.-40A>C and c.233+14G>A) were associated with obesity in Labrador dogs. The obtained results showed that the studied adipokine genes are highly polymorphic and two polymorphisms in the TNF gene may be considered as markers predisposing Labrador dogs to obesity.


Asunto(s)
Adipoquinas/genética , Perros/genética , Obesidad/genética , Polimorfismo de Nucleótido Simple , Animales , Cruzamiento , Genotipo , Haplotipos , Interleucina-6/genética , Resistina/genética , Análisis de Secuencia de ADN , Factor de Necrosis Tumoral alfa/genética
18.
Reprod Domest Anim ; 50(5): 776-83, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26303126

RESUMEN

To date, there is only scarce data on the evaluation of the prostate gland in dogs using computed tomography (CT). The aims of our study were to describe CT features of BPH in dogs and to determine the size of the prostate gland in healthy male dogs and dogs with benign prostatic hyperplasia (BPH) through CT. Additionally, we aimed to compare and establish the most useful parameters for CT measurements of the prostate in patients with BPH. The study population consisted of 20 healthy intact male dogs and 20 male intact dogs with confirmed BPH. Pre- and post-contrast CT studies were evaluated. The most common CT features in dogs with recognized BPH were symmetrical prostatomegaly and heterogeneity of the prostatic parenchyma. The mean prostatic density (D) was 56HU (±4.39) in pre-contrast CT images and 84HU (±8) in post-contrast images in dogs with BPH. The mean prostatic length (L) was 43.87 mm (±11), the mean width (W) amounted to 48.95 mm (±8.76) and the mean height (H) reached 44.9 mm (±9.48) in clinically affected patients. The mean ratios were: rL - 2,12 (±0.5); rW - 2.39 (±0.53) and rH - 2.16 (±0.39) in the BPH group. The prostate should be considered to be enlarged when rL exceeds 3.05; rW exceeds 3.38 and rH exceeds 2.94. Our findings indicated that CT is a useful tool in diagnosing prostate disorders, including BPH. The heterogeneity, density and ratios of prostatic length, width and height can be useful parameters in the diagnosis of BPH.


Asunto(s)
Enfermedades de los Perros/patología , Próstata/patología , Hiperplasia Prostática/patología , Tomografía Computarizada por Rayos X/veterinaria , Animales , Enfermedades de los Perros/diagnóstico por imagen , Perros , Masculino , Próstata/diagnóstico por imagen , Hiperplasia Prostática/diagnóstico por imagen
19.
Reprod Domest Anim ; 50(2): 344-348, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25611903

RESUMEN

An infertile Siamese female cat was subjected for clinical, histological, cytogenetic and molecular studies due to ambiguous external genitalia (vulva, vagina, rudimentary penis and scrotum-like structure) and masculine behaviour. An elevated oestrogen activity and a detectable level of testosterone were found. The cat underwent laparotomy. The gonads and the uterus were removed and subjected for histological studies, which showed ovaries with corpora lutea and a some primordial follicles. Chromosome studies of lymphocyte and fibroblast cultures, with the use of Giemsa staining, G-banding and whole X chromosome painting by fluorescence in situ hybridization, revealed pure X monosomy. Molecular analysis showed the absence of the SRY gene. Our study revealed for the first time that X monosomy in cats may be associated with virilization, in spite of the lack of the SRY gene.


Asunto(s)
Enfermedades de los Gatos/genética , Trastornos del Desarrollo Sexual/veterinaria , Aberraciones Cromosómicas Sexuales/veterinaria , Aneuploidia , Animales , Gatos , Femenino , Virilismo
20.
Reprod Domest Anim ; 49 Suppl 2: 8-15, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24947855

RESUMEN

The two most frequent prostatic diseases in dogs are benign prostatic hyperplasia (BPH) and prostatitis. Prostatitis requires prolonged antibiotic treatment. In acute prostatitis, the blood-prostate barrier is broken, thus facilitating the penetration of antibiotics, whereas in chronic prostatitis, the barrier prevents the penetration of many drugs into the gland. The selection of antibiotic agents is based on the sensitivity test and the drug's ability to penetrate into the gland. Many protocols for the treatment of BPH are available. In non-breeding dogs, surgical and optionally pharmacological castration by means of GnRH agonists may be performed. In breeding dogs, drugs retaining fertility are used. Recently, androgen receptor antagonistic treatment with osaterone acetate has been applied. Other drugs used for BPH treatment include progestagens, oestrogens, antioestrogens and 5α-reductase inhibitors. Some of these compounds may provoke severe side effects. The efficiency of GnRH antagonists used for the treatment of prostatic diseases, such as neoplasia and BPH, in humans has been recently investigated in dogs. This androgen deprivation therapy (ADT) is devoid of an initial exacerbation of androgen-dependent symptoms, which is typical for GnRH agonistic treatment. In many cases, BPH and prostatitis must be treated simultaneously as these conditions may develop in combination.


Asunto(s)
Antagonistas de Andrógenos/uso terapéutico , Antibacterianos/uso terapéutico , Enfermedades de los Perros/tratamiento farmacológico , Hormonas/uso terapéutico , Hiperplasia Prostática/veterinaria , Prostatitis/veterinaria , Animales , Perros , Masculino , Hiperplasia Prostática/tratamiento farmacológico , Prostatitis/tratamiento farmacológico
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