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1.
Theriogenology ; 138: 24-30, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31280182

RESUMO

Non-steroidal anti-inflammatory drugs (NSAIDs) are a therapeutic option for the treatment of inflammation. However, negative effects of non-selective NSAIDs for treatment of mares with endometritis have been described, including delayed uterine clearance and impairment of ovulations. Firocoxib is a specific cyclooxygenase-2 (COX-2) inhibitor and has the ability to act in the uterus of mares. We investigated the effects of firocoxib on ovulation rate, numbers of polymorphonuclear neutrophils (PMNs), and COX-2 protein levels in the endometrial tissue of susceptible mares after insemination. Two experiments were conducted. In experiment 1, twenty mares were evaluated in two consecutive estrous cycles broken into the following groups: Control - no pharmacological interference; Treatment - mares were treated with 0.2 mg/kg of firocoxib orally. The treatment began on the day of ovulation induction, and firocoxib was administered until one day after artificial insemination (AI). Ovulation was induced with 1 mg of deslorelin acetate and the mares were inseminated 24 h after the injection. Ovulation was confirmed 48 h after induction, and embryos were collected eight days after ovulation. Experiment 2: Nine mares susceptible to persistent mating-induced endometritis (PMIE) were artificially inseminated. The mares were examined with ultrasound and inseminated with fresh semen in two consecutive cycles, control and treated, in a cross-over study design. The amount of intrauterine fluid was measured, and endometrial samples were collected 24 h after AI. The number of PMNs was determined by endometrial cytology and biopsy, and COX-2 labeling in endometrial samples was evaluated by immunohistochemistry. Firocoxib treatment did not induce ovulatory failure or affect embryo recovery rate in Experiment 1. In Experiment 2, firocoxib treatment reduced inflammation after AI in mares as evidenced with results regarding PMN numbers/percentage and endometrial COX-2 staining. In conclusion, the proposed treatment with firocoxib reduced endometrial inflammation in mares susceptible to PMIE after breeding, with no adverse effects.

2.
Cryobiology ; 86: 58-64, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30557556

RESUMO

Boar semen cannot be immediately cryopreserved, it need be hold at 17 °C prior to cryopreservation, holding time has been used to improve cryopreserved boar semen, since holding time allows a prolonged interaction between spermatozoa and seminal plasma components. However, until now only few periods of holding time have been studied, and boar semen had been held at 17 °C for 24 h to facilitate its manufacture. Thus, this experiment aims to study the effect several holding time (0, 4, 8, 12, 24, 28 and 32 h) on boar spermatozoa post-thawed (PT) characteristics. Fifteen sperm-rich fractions of ejaculate were extended in Beltsville Thawing Solution and storage at 17 °C. After each holding time (0, 4, 8, 12, 24, 28 and 32 h), a sample was centrifuged, and sperm pellet was diluted in an extender composed of sugars, amino acids, buffers, 20% egg yolk (v/v), antibiotics, 2% glycerol as a cryoprotectant, and 2% methylformamide (v/v). Cryopreservation was performed with an automatic cryopreservation system. Cryopreserved boar semen was evaluated to spermatozoa kinetics, plasma and acrosomal membranes integrity, mitochondrial membrane potential, detection of superoxide anion, plasma membrane fluidity, and peroxidation. Twenty-four hours of holding increase total and progressive motility, rapid spermatozoa, and integrity of plasma and acrosome membranes. To mitochondrial membrane potential, 32 h is needed. However, holding time was not able to control the superoxide anion amount neither membrane lipid peroxidation, and had no effects on membrane fluidity. Thus, to reach the best results of PT boar semen the ideal holding time is 24 h.


Assuntos
Criopreservação/métodos , Análise do Sêmen , Preservação do Sêmen/métodos , Sêmen/fisiologia , Motilidade Espermática/fisiologia , Acrossomo/metabolismo , Animais , Membrana Celular/metabolismo , Crioprotetores/metabolismo , Crioprotetores/farmacologia , Masculino , Fluidez de Membrana , Potencial da Membrana Mitocondrial , Suínos , Fatores de Tempo
3.
Pesqui. vet. bras ; 38(9): 1726-1730, set. 2018. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-976505

RESUMO

To date, no studies have been performed evaluating the effect of boar spermatozoa concentration in 0.5mL freezing straws, leading us to examine this question. Each sperm-rich fraction of the ejaculate (n=25) was diluted at five different sperm concentrations (100, 200, 300, 600 and 800 x 106 spermatozoa/mL), packaged in 0.5mL straws, and subsequently frozen. After thawing, the sperm from all of treatment groups were analyzed to determine motility characteristics using a sperm class analyzer (SCA-CASA), and their plasma and acrosomal membrane integrity, mitochondrial membrane potential, sperm membrane lipid peroxidation and fluidity were analyzed by flow cytometry. An increase in spermatozoa concentration above 300x106 spermatozoa/mL in a 0.5mL straw impaired (p<0.05) the total and progressive motility, curvilinear velocity, straight-line velocity, linearity and beat cross frequency. However, the plasma and acrosomal membrane integrity, mitochondrial membrane potential, membrane lipid peroxidation and fluidity were not influenced (p>0.05) by high spermatozoa concentrations at freezing. Therefore, to increase spermatozoa survival and total and progressive motility after thawing, boar spermatozoa should be frozen at concentrations up to 300x106 spermatozoa/mL.(AU)


Até o momento, não foram realizados estudos que avaliassem o efeito da concentração de espermatozoides/mL em palhetas (0,5mL) para a criopreservação, levando-nos a analisar esta questão. Cada fração-rica do ejaculado (n=25) foi diluída em cinco diferentes concentrações de espermatozoides (100, 200, 300, 600 e 800x106 espermatozoides/mL), envasadas em palhetas de 0,5mL e posteriormente congeladas. Após a descongelação, os espermatozoides de todos os tratamentos foram avaliados a fim de determinar as características de motilidade usando um sistema de análise computadorizada dos espermatozoides (SCA-CASA). A integridade das membranas plasmática e acrosomal, o potencial de membrana mitocondrial, a peroxidação lipídica e a fluidez da membrana foram analisadas por citometria de fluxo. O aumento na concentração de espermatozoides acima de 300x106 espermatozoides/mL diminuiu (p<0,05) a motilidade total e progressiva, velocidade curvilínea, velocidade linear, linearidade e frequência de batimento. No entanto, a integridade da membrana plasmática e acrosomal, potencial de membrana mitocondrial, peroxidação lipídica e fluidez de membrana não foram influenciados (p>0,05) por altas concentrações de espermatozoides durante a criopreservação. Portanto, a fim de melhorar a sobrevivência dos espermatozoides suínos e a motilidade total e progressiva após a descongelação, os espermatozoides suínos devem ser congelados a concentrações não superiores a 300x106 espermatozoides/mL.(AU)


Assuntos
Animais , Suínos/embriologia , Criopreservação/veterinária , Análise do Sêmen/estatística & dados numéricos
4.
Anim Reprod Sci ; 192: 107-118, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29502896

RESUMO

This study aimed to evaluate the antioxidant properties of coenzyme Q10 (CoQ10) during cryopreservation of semen obtained from stallions having good and bad semen freezing ability (GFA vs. BFA, respectively). Forty ejaculates (n = 20 stallions) were split into five centrifugation and five freezing extenders containing different concentrations of CoQ10 (0, 25, 50, 75 and 100 µmols/L). If CoQ10 was added to the centrifugation extender, the freezing extender had no CoQ10 added; similarly, if CoQ10 was added to the freezing extender, the centrifugation extender had no CoQ10. Semen cryopreserved on extenders containing no CoQ10 served as the control. After post-thaw total sperm motility (TM) assessments, the stallions were classified as GFA (i.e., decrease of ≤25% in TM, n = 7) or BFA (i.e., decrease of ≥40% in TM, n = 5). Stallions not fitting (n = 8) this enrollment criteria had samples discarded. After that, two straws for each extender were thawed at 37 °C for 30 s; one straw was immediately used for evaluation of sperm kinetics, plasma membrane integrity, non-capacitated spermatozoa, reactive oxygen species production, mitochondrial activity and lipid peroxidation. The second straw was kept at 37 °C for 30 min and subjected to the same assessments. Expectedly, sperm motility parameters were significantly lower for stallions with BFA. There were no effects of CoQ10 concentration or time for all parameters evaluated in the group with GFA when compared with the control extender (p > 0.05), except lipid peroxidation (p < 0.05). However, stallions with BFA had improved sperm parameters for samples processed with extenders containing CoQ10 (particularly 75 µmols/L) (p < 0.05), except for the reactive oxygen species production and mitochondrial potential (T0) in which there were no differences between the groups (p > 0.05). In summary, 75 µmols/L appears to be the optimal dose of Co-Q10, particularly, when added to the centrifugation extender.


Assuntos
Criopreservação/veterinária , Cavalos/fisiologia , Preservação do Sêmen/veterinária , Ubiquinona/análogos & derivados , Animais , Congelamento , Masculino , Ubiquinona/farmacologia
5.
Anim Reprod Sci ; 190: 1-9, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29373173

RESUMO

This study compared hormone treatments given to mares during anestrus, spring transition, and different stages of the estrous cycle, by assessing uterine features and pregnancy rates after embryo transfer (ET). Embryo recipient mares (n = 160) were equally arranged as follows: G1-spontaneous ovulation (control), G2-anestrus, G3-spring transition, G4-early estrus, G5-estrus, G6-diestrus, G7-early diestrus treated with a dose of dinoprost, and G8-early diestrus treated with two doses of dinoprost. At treatment initiation (Day-4), G2-7 were given dinoprost and estradiol-17ß, thereafter, estradiol-17ß was repeated on Days-3,-2, and -1. On Day0, mares received long-acting altrenogest. Then, each mare had one ET performed from Day + 3 to Day + 8 after altrenogest. Immediately before the ET, mares received a boost of altrenogest and had uterine features assessed. Pregnant mares on each of the checks (by 7, 30, 60, and 120d after ET) were maintained on weekly injections of LA-P4 until 120d. G8 received similar management, but dinoprost was repeated on Day-3. G1-G6 and G8 displayed uterine edema and satisfactory pregnancy rates ≥65%. Repeating dinoprost to G8 likely ensured proper luteolysis and response to estrogen as determined by higher uterine edema scores and pregnancy rates than G7 (p < .05). Our results were consistent with previous studies and other successful commercial ET programs (except G7), thus, demonstrating the usefulness of the hormone treatments described herein to synchronize embryo recipient mares with donor mares. Thus, we foresee that other groups may use the strategies described herein for the management of embryo recipient mares.


Assuntos
Dinoprosta/farmacologia , Transferência Embrionária/veterinária , Estradiol/farmacologia , Cavalos/fisiologia , Acetato de Trembolona/análogos & derivados , Anabolizantes/administração & dosagem , Anabolizantes/farmacologia , Animais , Dinoprosta/administração & dosagem , Estradiol/administração & dosagem , Estrogênios/administração & dosagem , Estrogênios/farmacologia , Ciclo Estral , Sincronização do Estro , Feminino , Ocitócicos/administração & dosagem , Ocitócicos/farmacologia , Gravidez , Taxa de Gravidez , Progesterona/farmacologia , Progestinas/farmacologia , Acetato de Trembolona/administração & dosagem , Acetato de Trembolona/farmacologia
6.
J Equine Vet Sci ; 64: 89-95, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-30973159

RESUMO

The aim of this report is to describe a new methodology to successfully treat stallions diagnosed with urethral rent. Four stallions of ages ranging from 7 to 12 years (median 9) with hemospermia were admitted for clinical evaluation, breeding soundness examination, and urethroscopy for inspection of the urethra and vesicular glands. Once the presence of urethral rent was identified and/or other sources of hemorrhage were excluded, a topical treatment was performed with 4% Policresulen solution (Albocresil). The treatment was carried out by infusing 100 mL of the solution into the lumen of the urethra through a catheter placed up to the region of the ischial arch. This procedure was repeated once daily, or at 48 hours intervals, resulting in a total of 4-7 infusions. In all cases, chemical cauterization was efficient in the healing of the urethral rent. However, due to masturbation during treatment, one animal did not completely heal, and the treatment with the Policresulen was prolonged. It is believed that the low pH of the solution resulted in urethritis, which was treated with systemic therapy of antibiotic and anti-inflammatory nonsteroidal. Topical treatment with 4% Policresulen was found to be efficient in the chemical cauterization of urethral rent in stallions. This treatment was efficient, practical, less invasive, and less costly than the alternative of surgical methods, which are more invasive and require longer recovery time of the animal. However, sexual rest and the elimination of sexual stimuli from the environment are essential management in association with this therapeutic method.

7.
Theriogenology ; 104: 120-126, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28822903

RESUMO

Persistent mating-induced endometritis (PMIE) results in decreased fertility in horses, thereby causing a significant impact in the horse market. Platelet-rich plasma (PRP), a modulator of the inflammatory response, has been largely used in veterinary medicine. Here, we investigated the effects of PRP on uterine inflammation, conception rate, endometrial polymorphonuclear neutrophil (PMN) migration, and COX-2 protein levels in the endometrial tissue. Thirteen PMIE-susceptible mares were used for artificial insemination (AI). The mares were inseminated with fresh semen in three consecutive cycles in a cross-over study design. The following cycle classifications were used: control cycle, no pharmacological interference; pre-AI, 20 mL of PRP was infused 24 h before AI; and post-AI, 20 mL of PRP was infused four h after AI. Follicular dynamics were monitored daily by transrectal ultrasound. When a follicle larger than 35 mm was detected, ovulation was induced with deslorelin acetate (1 mg, im). AI was performed 24 h after ovulation induction. Intrauterine fluid (FLU) was evaluated by ultrasonography before and 24 h after AI. PMNs in uterine cytology (CYT) and biopsy (HIS) were also observed before and 24 h after AI. Pregnancy was determined within 14 days after ovulation. Number of COX-2 positive cells was evaluated by immunohistochemistry. Both PRP treatments resulted in a decrease of PMNs in the CYT after breeding when compared to controls. FLU did not differ between cycles; however, the conception rates were significantly higher in the PRP mares. Mares positive for endometritis decreased in both treatment groups, and a more intense positive COX-2 labeling was observed in the control group when compared to the two treatment groups. In conclusion, PRP beneficially reduces inflammatory response in PMIE mares independent of when treatments were administered, thus increasing chances of successful pregnancy.


Assuntos
Endometriose/veterinária , Endométrio/metabolismo , Doenças dos Cavalos/prevenção & controle , Cavalos/fisiologia , Inseminação Artificial/veterinária , Leucócitos/efeitos dos fármacos , Animais , Movimento Celular/fisiologia , Endometriose/prevenção & controle , Endométrio/efeitos dos fármacos , Feminino , Leucócitos/fisiologia , Plasma Rico em Plaquetas , Gravidez , Taxa de Gravidez
8.
Theriogenology ; 86(2): 516-22, 2016 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-27020400

RESUMO

Degenerative changes of the endometrium are directly related to age and fertility in mares. Chronic degenerative endometritis (CDE) is correlated with uterine fluid retention and reduced ability to clear uterine inflammation. Recent research in the areas of equine surgery and sports medicine has shown that platelet-rich plasma (PRP) treatment acts as an immunomodulator of the inflammatory response. Therefore, the aim of this study was to determine if the uterine infusion of PRP could modulate the local inflammatory response and modify the intrauterine NO concentrations after artificial insemination (AI) in both normal mares and those with CDE. Thirteen mares with endometrium classified as grade III on the histology (mares with CDE) and eight mares with endometrial histological classification I or II-a normal mares were selected to investigate the effect of PRP therapy. The mares were inseminated with fresh semen in two consecutive cycles in a crossover study design. Thereby, each mare served as its own control and the treatment was performed with intrauterine PRP infusion four hours after AI. The percentage of neutrophils in uterine cytology (CIT, %), uterine fluid accumulation observed on ultrasonography (FLU, mm) and nitric oxide concentration of uterine fluid (NO, µM) were analyzed before and 24 hours after AI. The results reported that mares with CDE (CIT, 68.3 ± 3.27, FLU, 10.7 ± 1.61) have a higher (P < 0.05) intrauterine inflammatory response after AI than normal mares (CIT, 24.4 ± 3.56, FLU, 0), but NO concentrations did not differ (P > 0.05) between categories of mares. In treated cycles with PRP, the intrauterine inflammatory response decrease (P < 0.05) in CDE mares (CDE: CIT, 31.4 ± 6.48, FLU, 5.5 ± 1.28; normal mares: CIT, 13.5 ± 4.31, FLU, 0) when compared with nontreated cycle (CDE: CIT, 68.3 ± 3.27, FLU, 10.7 ± 1.61; NM: CIT, 24.4 ± 3.56, FLU, 0), but did not modify NO concentrations in uterine fluid. Thus, we can conclude that PRP was effective in modulating the exacerbated uterine inflammatory response to semen in mares with CDE but did not reduce NO concentrations in intrauterine fluid.


Assuntos
Endometrite/veterinária , Doenças dos Cavalos/terapia , Inflamação/metabolismo , Plasma Rico em Plaquetas , Animais , Estudos de Casos e Controles , Endometrite/terapia , Feminino , Cavalos
9.
Anim Reprod Sci ; 161: 58-63, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26315896

RESUMO

The objective of this study was to evaluate the effects of two different egg yolk extenders incubated with or without Sperm Talp on the motility and plasma membrane integrity of cryopreserved bovine epididymal spermatozoa after freezing. Twenty-five testicles with epididymides from mature bulls were collected at the abattoir. Epididymal sperm recovery was performed by retrograde flushing using a skim milk-extender (Botu-Semen™). After recovery, sperm were incubated either without or with Sperm Talp and then submitted to centrifugation. For the freezing process, half of the testes were processed with Tris egg yolk extender, and half were processed with Botu-Bov™ egg yolk extender. Samples incubated in Sperm Talp exhibited better results than epididymal spermatozoa that were incubated without Sperm Talp (p<0.05). Both Botu-Bov™ and Tris could be utilised to freeze sperm from the bovine epididymides if the sperm were previously incubated with Sperm Talp. The extenders examined in this work did not differ in their effect on plasma membrane integrity after freezing.


Assuntos
Criopreservação/veterinária , Espermatozoides/metabolismo , Animais , Bovinos , Criopreservação/métodos , Meios de Cultura , Epididimo/citologia , Masculino , Espermatozoides/efeitos dos fármacos
10.
Theriogenology ; 80(7): 730-7, 2013 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-23932173

RESUMO

Cryopreservation of spermatozoa is a pivotal tool in assisted reproduction, and studies aiming to establish optimal freezing/thawing protocols are essential to enhance sperm survival. The objectives of the present study were to (1) compare the cryoprotective efficiency of three different glycerol concentrations (3%, 5%, and 7%) on the basis of post-thaw sperm quality and (2) investigate whether the incidence of morphologically abnormal sperm in fresh samples is related to cryodamage sensitivity. Semen was collected from six tomcats using an artificial vagina (total 18 ejaculates). Each ejaculate was diluted using Tris-egg yolk-based extender (TEY), evaluated, equally divided into three aliquots, and rediluted using TEY with and without glycerol to achieve final concentrations of 3%, 5%, and 7%. Samples were loaded into 0.25 mL straws, equilibrated for 60 minutes at 5 °C, frozen, and then thawed at 46 °C for 12 seconds. Fresh and frozen-thawed samples were evaluated for sperm motion parameters (computer-assisted sperm analysis), plasma membrane integrity (PMI; propidium iodide and carboxyfluorescein diacetate), and DNA integrity (acridine orange). Plasma and acrosomal membrane integrity were assessed by flow cytometry (propidium iodide and fluorescein isothiocyanate-conjugated pea (Pisum sativum) agglutinin) immediately after thawing. Sperm motion parameters were also evaluated at 30 and 60 minutes of postincubation. For all treatment groups, cryopreservation significantly impaired the PMI and sperm motion parameters, except for straightness and amplitude of lateral head displacement. DNA integrity showed a slight reduction (P < 0.05) when 3% glycerol was used. The percentage of total motility, progressive motility, and rapid spermatozoa were significantly lower immediately after thawing and up to 60 minutes of incubation for the 3% glycerol group when compared with 5% and 7%. No difference (P > 0.05) was found for PMI, acrosome integrity, and DNA integrity among post-thaw groups. However, higher (P < 0.05) incidence of viable cells with reacted acrosome and dead cells with intact acrosome were observed with 7% and 3% glycerol, respectively. Percentage of morphologically abnormal spermatozoa in fresh sample was positively correlated with PMI only in the 3% glycerol group and negatively correlated with sperm motility in the 5% and 7% groups. In conclusion, the final concentration of 5% glycerol offered better cryoprotective effect for ejaculated cat sperm, and the relationship found between prefreezing sperm morphology and post-thaw sperm quality showed to be dependent on final glycerol concentration.


Assuntos
Crioprotetores/farmacologia , Glicerol/farmacologia , Espermatozoides/efeitos dos fármacos , Acrossomo/efeitos dos fármacos , Animais , Gatos , Criopreservação/métodos , Criopreservação/veterinária , Masculino , Análise do Sêmen/veterinária
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