RESUMO
Vitrification is essential for successful tissue cryopreservation and biobanking in wild cats. This study aimed to compare different methods of vitrification (Ovarian Tissue Cryosystem-OTC, Straws-STW, and Solid Surface vitrification-SSV) for testicular fragment vitrification in tom cats. Testicular fragments were recovered from five adult tom cats and subjected to equilibrium vitrification using different cryovials and methods under the same conditions of vitrification solutions and cryoprotectants. The efficiencies of the methods were evaluated using histological analysis of spermatogonia and Sertoli cell nuclei, seminiferous tubular basement membrane detachment, and the gonadal epithelium shrinkage score scale. Cell viability was assessed using Hoechst PI and Terminal deoxynucleotidyl transferase nick end labeling (TUNEL) assay. The results showed that OTC is an effective vitrification method for maintaining the distinction between spermatogonia and Sertoli cells. OTC was similar to the control for basal membrane detachment parameters (p = 0.05). Epithelial shrinkage was low in the SSV group, which showed the highest percentage of viable cells among the vitrified groups (p = 0.0023). The OTC and SSV vitrification methods were statistically similar in terms of the percentage of TUNEL-positive cells (p = 0.05). Therefore, OTC and SSV provide favorable conditions for maintaining viable cat testicular tissue cells after vitrification.
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This study evaluated the effects of different concentrations (10, 20, or 40 µM) of eugenol (EUG 10, EUG 20, or EUG 40), ascorbic acid (50 µg/mL; AA) or anethole (300 µg/mL; ANE 300) on the in-vitro survival and development of goat preantral follicles and oxidative stress in the cultured ovarian tissue. Ovarian fragments from five goats were cultured for 1 or 7 days in Alpha Minimum Essential Medium (α-MEM+) supplemented or not with AA, ANE 300, EUG 10, EUG 20 or EUG 40. On day 7 of culture, when compared to MEM, the addition of EUG 40 had increased the rate of follicular development, as observed by a decrease in the proportion of primordial follicles alongside with an increase in the rate of normally developing follicles. Furthermore, EUG 40 significantly increased both follicular and oocyte diameters. Subsequently, ovarian fragments from three goats were cultured for 1 or 7 days in α-MEM+ supplemented or not with AA, ANE 300 or EUG 40. All tested antioxidants, except ANE 300, were able to significantly decrease the levels of reactive oxygen species in the ovarian tissue, but EUG 40 could most efficiently neutralize free radicals. All ovarian tissues cultured in the presence of antioxidants, especially EUG 40, presented a significant decrease in H3K4me3 labeling, indicating a silencing of genes that play a role in the inhibition of follicular activation and apoptosis induction. When compared to cultured control tissues, both EUG 40 and ANE 300 significantly increased the intensity of calreticulin labeling in growing follicles. The mRNA relative expression of ERP29 and KDM3A was significantly increased when the culture medium was supplemented with EUG 40, indicating a response to ER stress experienced during culture. In conclusion, EUG 40 improved in-vitro follicle survival, activation and development and decreased ROS production, ER stress and histone lysine methylation in goat ovarian tissue.
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This study aimed to describe the viability of domestic feline spermatozoa after epididymal tail vitrification. For this, 10 pairs of testis-epididymis complexes were used. The epididymal tails were vitrified using the solid-surface vitrification (SSV) method, in which two vitrification media containing ethylene glycol (EG) 40% or glycerol (GLY) 40% were tested. Vitrification with the presence of EG resulted in better results for all sperm motility parameters (motility, vigour and CASA) compared with GLY (P < 0.05). There were no statistical differences for sperm viability and acrosome integrity, plasma membrane integrity, or overall health of morphologically normal sperm before or after vitrification among experimental groups. In conclusion, epididymal tail vitrification appears to be a suitable method for long-term storage of cat sperm, especially if the procedure is performed with EG as the cryoprotectant.
Assuntos
Preservação do Sêmen , Vitrificação , Animais , Gatos , Criopreservação/veterinária , Crioprotetores/farmacologia , Epididimo , Masculino , Preservação do Sêmen/veterinária , Motilidade dos Espermatozoides , EspermatozoidesRESUMO
Two farms applying reproductive technology for the Nellore beef cattle were selected. Both farms had the same technology programme of oestrous synchronization and embryo transfer, but management was different, especially regarding twins pregnancies. In the present study, we followed the farms from the moment of oestrous synchronization, embryo transfer (two per cow), until delivery and first care of the calves. In farm A, cows presenting twin pregnancies (5 from 13) were submitted to delivery induction, as well as calves and cows were monitored after birth. In farm B, such management was not followed with the twin pregnant cows (31 from 49). In both farms, freemartinism was detected, but this was not a problem as none of the animals would be selected for breeding. No dystocia was observed in farm A, while 48% of the twin pregnancies in farm B ended up in dystocia. Furthermore, the mortality rate of new-born calves in farm A was 10%, while in farm B it reached 32%. Although twin pregnancies remain a concern, we showed here that proper management during and after delivery minimizes animal and economic losses.
Assuntos
Transferência Embrionária , Resultado da Gravidez/veterinária , Prenhez , Gravidez Múltipla , Animais , Animais Recém-Nascidos , Bovinos , Ciclo Estral , Fazendas , Feminino , Fertilização in vitro , Freemartinismo/genética , Trabalho de Parto Induzido/veterinária , Masculino , Mortalidade , Gravidez , Taxa de GravidezRESUMO
Saimiri collinsi is used as an animal model in biotechnology research for conservation of species from the genus Saimiri. However, the development of biotechnologies depends on a proper knowledge of the sperm morphology to understand the basic aspects of sperm physiology, as potential male fertility depends on different cellular sperm structures. With this purpose, this study characterized the micromorphological and ultrastructural characteristics of squirrel monkeys (Saimiri collinsi) sperm using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). SEM electromyography revealed that a normal Saimiri collinsi sperm measures 71.7 ± 0.7 µm with lateral tail insertion, a paddle-shaped flattened head and an acrosome occupying most of the head. TEM also showed that the middle piece is characterized by a central 9 + 2 microtubule axoneme surrounded by nine dense fibres, and that the mitochondria were juxtaposed, forming the mitochondrial sheath. Here we provide the first micromorphological and ultrastructure description of S. collinsi sperm.
Assuntos
Acrossomo/ultraestrutura , Cabeça do Espermatozoide/ultraestrutura , Cauda do Espermatozoide/ultraestrutura , Espermatozoides/ultraestrutura , Acrossomo/fisiologia , Animais , Axonema/ultraestrutura , Membrana Celular/ultraestrutura , Núcleo Celular/ultraestrutura , Humanos , Masculino , Microscopia Eletrônica de Varredura/métodos , Microscopia Eletrônica de Transmissão/métodos , Mitocôndrias/ultraestrutura , Sêmen/citologia , Cabeça do Espermatozoide/fisiologia , Motilidade dos Espermatozoides , Cauda do Espermatozoide/fisiologia , Espermatozoides/fisiologiaRESUMO
The aim of the present study was to establish a protocol for solid surface vitrification of peccary ovarian tissue by using different cryoprotectants. Ovarian pairs from five adult females were fragmented and two fragments (fresh control group) were immediately subjected to morphological evaluation using classical histology, transmission electron microscopy, and viability analysis using fluorescent probes. The remaining fragments (n = 18) were vitrified using a solid surface method with different concentrations (3 or 6 M) of ethylene glycol (EG), dimethyl sulfoxide (DMSO) or dimethyl formamide (DMF). After 2 weeks, samples were re-warmed and evaluated. A decrease in the percentage of morphologically normal preantral follicles (PFs) was verified for all the groups in comparison to the fresh control (92.0 ± 2.8%); however, if only the primordial follicles are considered, the most effective preservation (P < 0.05) was achieved with the use of EG at 3 M (74.2±7.3%) or DMSO at 6 M (75.0 ± 4.2%). Ultrastructural analysis indicated there were well-preserved PFs in all the groups evaluated, having well-defined membranes, a few vacuoles, and organelles that were uniformly distributed throughout the cytoplasm, mainly round and elongated mitochondria in close association with lipid droplets. Viability was preserved (P < 0.05) with the use of EG at 3 (97%) or 6 (97%) M, DMSO at 3 (100%), and DMF at 6 (97%) M. Solid surface vitrification, therefore, is an effective method for conservation of peccary female germplasm, especially with the use of EG at 3 M, which was highly effective for preservation of both the morphology and viability of PFs.
Assuntos
Artiodáctilos/fisiologia , Crioprotetores/farmacologia , Ovário/fisiologia , Preservação de Tecido/veterinária , Vitrificação/efeitos dos fármacos , Animais , Sobrevivência Celular , FemininoRESUMO
Bone fractures in birds are usually diagnosed with the aid of traditional radiography. However, this technique remains limited because of the difficulties associating this examination with real-time procedures. The aim of this study was to describe the use of B-mode ultrasound to assess the long bones of two avian orders. For the study, we used carcases of birds from the orders Falconiformes (n = 9) and Strigiformes (n = 12), with weights ranging from 108 to 1020â g. An ultrasound device with a 5-12â MHz linear probe was employed to produce images of the long bones (humerus, radius, ulna, femur and tibiotarsus). Ultrasound (US) measurements and physical measurements using a caliper were applied to compare the diameter of the bones. Images were also recorded from the US examination performed in two live patients attending the hospital with suspected bone fractures. No statistical difference was found between the two methods of measurement in carcases weighing up to 267â g (P > 0.01). The US examination provided relevant clinical information about the bone cortex and assisted in real-time surgical procedures. RESEARCH HIGHLIGHTS Long bones of Falconiformes and Strigiformes birds were assessed with B-mode ultrasound. Ultrasound analysis is a relevant tool in clinical orthopaedics for avian species. Ultrasound of the bone might be applied for monitoring of healing processes.
Assuntos
Doenças das Aves/diagnóstico por imagem , Fraturas Ósseas/veterinária , Animais , Osso e Ossos/diagnóstico por imagem , Falconiformes , Fraturas Ósseas/diagnóstico por imagem , Estrigiformes , Ultrassonografia/veterináriaRESUMO
We aimed to evaluate the effect of three extracellular cryoprotectants on the morphology of vitrified feline preantral follicles. Feline ovarian fragments (0.5 × 2 × 2 mm) collected from five domestic adult cats subjected to ovariohysterectomy for routine castration were vitrified with ethylene glycol (EG) 40% combined or not with sucrose (0.1 or 0.5 M), trehalose (0.1 or 0.5 M), or raffinose (0.1 M). After vitrification using the solid-surface method and warming of the tissues, cryoprotectants were washed out of the ovarian tissues, which were fixed for histological analysis. The percentages of normal follicles were similar to the control (fresh) (62.9 ± 4.1%) only for tissues exposed and cryopreserved with EG + trehalose at concentrations of 0.1 (35.8 ± 8.3%) and 0.5 M (33.4 ± 5.4%). All the other sugars decreased the percentages of morphologically normal follicles as compared to the control group and the trehalose groups. Based on the results of the present study, we recommend the use of trehalose as the extracellular cryoprotectant for the vitrification of feline ovarian tissue.
Assuntos
Criopreservação/métodos , Crioprotetores/farmacologia , Preservação de Órgãos/métodos , Folículo Ovariano/efeitos dos fármacos , Vitrificação , Animais , Gatos , Etilenoglicol/farmacologia , Feminino , Rafinose/farmacologia , Sacarose/farmacologia , Trealose/farmacologiaRESUMO
The objective of this study was to assess the influence of nutritional regimens such as adequate feeding, restricted feeding, and underfeeding-refeeding on the follicle growth and development from caprine ovaries. Goats were divided into three different groups (n = 5 per group). For 24 weeks, goats received elephant grass plus concentrate to provide 1.5 (n = 5) and 0.72 (n = 10) times the energy requirements for maintenance of live weight. Underfed goats were subsequently refed for 6 weeks with the diet of the nourished group (1.5 times the energetic requirements of maintenance). Follicular morphology and morphometry, as well as granulosa cells mitotic index were assessed. Ovarian follicles were classified as small or large preantral follicles, or as small or large antral follicles. Ovarian volume was smaller in animals from both underfed and refed groups than in those animals from fed group. Although no difference in the total number of normal follicles was observed among the nutritional groups, underfed animals presented higher percentages of atretic preantral and small antral follicles when compared with fed animals. Large antral follicles from underfed and refed goats presented a lower mitotic index when compared with fed ones. In conclusion, ovaries from goats challenged with prolonged undernutrition will be functionally compromised, which is characterized by atresia of preantral and small antral follicles and decreased mitotic index of large antral follicles. Refeeding those animals will not recover ovarian function to a same level experienced by goats fed a diet with adequate energy requirements.
Assuntos
Fenômenos Fisiológicos da Nutrição Animal/fisiologia , Cabras/fisiologia , Índice Mitótico , Folículo Ovariano/fisiologia , Ração Animal , Animais , Contagem de Células , Feminino , Células da Granulosa/fisiologia , Folículo Ovariano/citologia , Ovário/citologia , Ovário/fisiologiaRESUMO
We describe morphological and morphometrical characteristics of preantral ovarian follicles from three recently recognized Saimiri species: S. macrodon, S. cassiquiarensis and S. vanzolinii; the last one a threatened species. Ovaries from four adult monkeys were evaluated: one pair from a pregnant S. macrodon, two ovarian pairs from S. cassiquiarensis females (one of them pregnant), and one left ovary from a senile S. vanzolinii, applying classical histology. Follicular preantral population was quantified and morphology and morphometry of primordial, primary and secondary follicles were evaluated. Follicular preantral population varied among species, being 347,153 in the ovaries of the S. macrodon, 270,342 and 278,376 in the ovaries of both adult non-pregnant and pregnant S. cassiquiarensis females, and 28,149 in the ovary from a senile S. vanzolinii. Most follicles were at primordial or transition stages, except for the senile S. vanzolinii female, which presented the lowest percentages of primordial and transition follicles when compared with primary and secondary ones. Most preantral follicles (>70%) were morphologically normal in the ovaries from all studied S. macrodon and S. cassiquiarensis females, but the ovary of the senile S. vanzolinii female presented a significant decrease in the percentage of normal follicles (primordial: 61%, transition: 52%, primary: 54%, and secondary: 48%). In general, follicular diameter increased significantly from primordial to transition, and subsequently from primary to secondary follicles.
Assuntos
Folículo Ovariano/anatomia & histologia , Saimiri/anatomia & histologia , Animais , Brasil , Feminino , Microscopia Eletrônica de Varredura , Oócitos/citologia , Oócitos/fisiologia , Folículo Ovariano/fisiologia , Ovário/anatomia & histologia , Ovário/fisiologia , Gravidez , Saimiri/fisiologia , Especificidade da EspécieRESUMO
We aimed to evaluate the effect of supplementation of ACP-118® extender with the antioxidant catalase (10 and 50 µg/ml) on Sapajus apella sperm motility, vigour, and plasma membrane integrity during the processes of seminal liquefaction, cooling, and freezing. Catalase did not affect any of the evaluated parameters after semen dilution or cooling. Cryopreserved sperm in the presence of 50 µg/ml catalase presented a plasma membrane integrity similar to that fresh sperm, however.
Assuntos
Catalase/metabolismo , Membrana Celular/química , Crioprotetores/farmacologia , Congelamento , Sêmen/efeitos dos fármacos , Motilidade dos Espermatozoides/efeitos dos fármacos , Animais , Cebus , Membrana Celular/efeitos dos fármacos , Criopreservação/métodos , Criopreservação/veterinária , Masculino , Preservação do Sêmen/métodos , Preservação do Sêmen/veterináriaRESUMO
Ovarian agenesis is an unusual anomaly with traumatic or congenital origin. In the present case report, we describe our findings in a senile S. vanzolinii female. As this neotropical primate species is listed as vulnerable, with limited geographic distribution in the Brazilian Amazonia, ovarian agenesis may be an important finding to be reported.
Assuntos
Disgenesia Gonadal/veterinária , Ovário/anormalidades , Saimiri/anormalidades , Animais , Brasil , Espécies em Perigo de Extinção , Feminino , Disgenesia Gonadal/diagnóstico por imagemRESUMO
In the present study, we aimed to assess the influence of different social contexts on the seminal coagulation and sperm quality in captive tufted capuchin monkeys. For this, males were housed either individually, in mixed-sex groups (with females), or in male-only groups. Monkeys were housed in cages and each cage type (i.e., individual or group cage) was placed in a different room. Forty-one males were subjected to semen collection by rectal electroejaculation. The degree of seminal coagulation was determined on a scale of I-IV. Seminal volume, sperm concentration, sperm motility, vigor, and plasma membrane integrity were evaluated for all ejaculate samples. All ejaculates collected showed degrees of coagulation between II and IV, where the majority presented coagulation degree IV, when collected from animals housed in groups. No statistical differences among percentages of coagula degree when samples were collected from males housed individually. Animals housed in group cages (male-only groups and mixed-sex groups) showed a significantly higher percentage of ejaculates at degree IV than males housed individually. Seminal volume was not affected by the coagula degree but by the housing system, where animals housed individually showed the highest volume (543 µl) when compared with those animals from male (273 µl) and mixed-sex (318 µl) groups. No differences were observed in semen volume when comparing male-only groups with mixed-sex groups. Sperm motility was affected by both housing system and coagula degree. Samples with coagula degree IV from animals housed individually showed the highest (72%) sperm motility percentages. Sperm plasma membrane integrity was lower when samples were presenting coagula degree II + III and collected from male- (17%) or mixed-sex (23%) groups. However, this housing system effect was not observed when sperm was obtained from coagula degree IV semen. Sperm vigor was neither affect by housing system or coagula degree.
Assuntos
Cebus , Comportamento Social , Contagem de Espermatozoides , Motilidade dos Espermatozoides , Animais , Feminino , Masculino , Sêmen , EspermatozoidesRESUMO
Sperm morphometry can be applied to identify different animal groups and species and to evaluate sperm quality. Furthermore, knowledge on species-specific differences will help to enhance biological information, as well as to develop efficient reproductive technologies. The aims in the present study were to describe sperm morphometry from the recently characterized species S. collinsi and S. vanzolinii, to verify if the morphometric sperm patterns are similar or different between both species, and to determine if the sperm morphometry is affected by the levels of sperm defects using the S. collinsi as a model. Semen was collected from S. collinsi (n = 10) and S. vanzolinii (n = 2) monkeys, and sperm was submitted to morphological analysis. From the 10 samples from S. collinsi, five presented sperm of poor quality and two subgroups were formed for this species, i.e. high and poor quality sperm. Data on sperm motility and vigour were analysed, as well morphometric parameters on sperm head and tail. It was observed the normal morphometry was correlated with high quality sperm. Poor quality sperm presented smaller and 7% more ellipticity in their head, when compared with high quality sperm. Sperm from S. vanzolinii presented larger head than those from S. collinsi, but tail lengths were similar. Sperm morphometry can be used as a complementary tool to predict sperm motility and vigour for the S. collinsi species, and S. collinsi appear as a suitable model for S. vanzolinii.
Assuntos
Microscopia/métodos , Saimiri/anatomia & histologia , Sêmen/fisiologia , Motilidade dos Espermatozoides/fisiologia , Espermatozoides/química , Animais , Processamento de Imagem Assistida por Computador , Masculino , Filogenia , Saimiri/classificação , Saimiri/fisiologia , Especificidade da EspécieRESUMO
The aim of this study was to investigate the effects of different media with or without phenol red or the antioxidant trolox on the successful vitrification of feline ovarian tissue. In a first experiment, ovarian cortical pieces from three cats were vitrified in solutions of Roswell Park Memorial Institute (RPMI)-1640 medium, Minimum Essential Medium, Dulbecco's modified Eagle's medium, or Tissue Culture Medium 199 as basic medium, supplemented or not with 50 µM of trolox, all containing 40% ethylene glycol (EG) and 1 M of sucrose. RPMI-1640 (phenol red-free) without trolox was the only medium that preserved the percentage of morphologically normal preantral follicles similar to control (80%). The main difference between RPMI-1640 and the other media was the absence of phenol red and CaCl2. In a second experiment, ovarian cortical pieces from three cats were vitrified in a solution containing RPMI-1640 as basic medium, 40% EG, 1 M of sucrose, supplemented or not with phenol red or CaCl2 alone, or in combination. It was observed that phenol red supplementation led to follicular degeneration. Finally, to evaluate the interaction between phenol red and the cryoprotectant agent (i.e., EG), ovarian tissue was exposed to RPMI-1640 supplemented with phenol red and EG at different concentrations (10%, 20%, or 40%). There was an inverse relationship between EG concentration and free phenol red in the medium after exposure. It is suggested that vitrification of feline ovarian tissue should be performed in a phenol red-free medium. Medium supplementation with 50 µM of trolox was deleterious for follicular morphology.
Assuntos
Crioprotetores/efeitos adversos , Ovário/efeitos dos fármacos , Fenolsulfonaftaleína/efeitos adversos , Preservação Biológica/métodos , Vitrificação/efeitos dos fármacos , Animais , Gatos , FemininoRESUMO
Antibiotics are widely used for the treatment of dairy cows and the residues of these drugs may remain in milk and dairy products, which can be a potential threat to human health. Exposure to low levels of antibiotics is considered a public health problem as this may result in the development of resistant strains of human bacteria. The presence of antibiotic residues (AR) in milk is also a problem for the dairy industry as they can inhibit the growth of lactic bacteria. According to Brazilian legislation, antimicrobials should be used in accordance with Good Farming Practices. However, recent studies have reported contamination in milk marketed in the country. This work aimed to review studies, published over the last 10 years, which describe AR in milk marketed in Brazil. The Maximum Residue Limit, the methods for quantification of AR and the results of published studies by authors and government agencies are discussed.
Los antibióticos se utilizan ampliamente para el tratamiento de las vacas lecheras y los residuos de estos medicamentos pueden permanecer en la leche y los productos lácteos, los que pueden ser una amenaza potencial para la salud humana. La exposición a bajos niveles de antibióticos se considera un problema de salud pública ya que pueden resultar en el desarrollo de cepas resistentes de bacterias humanos. De acuerdo con la legislación brasileña, los antimicrobianos deben ser utilizados de acuerdo con las Buenas Prácticas Agrícolas. Sin embargo, estudios recientes han informado de la contaminación de la leche comercializada en el país. Este trabajo tuvo como objetivo revisar los estudios publicados en los últimos 10 años, que describen residuos de antimicrobianos en la leche comercializada en Brasil. Se discuten los límites máximos de residuos, los métodos para la cuantificación y los resultados de los estudios publicados por autores y agencias gubernamentales.
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Humanos , Qualidade dos Alimentos , Impactos da Poluição na Saúde , Indústria de Laticínios , Leite , Anti-InfecciososRESUMO
There is a paucity of efficient cryopreservation protocols for primordial follicles enclosed in the ovarian tissue from non-human primates (NHP), in special New World primates. Our objective was to establish an optimal procedure for the recovery of ovarian biopsies from capuchin monkeys. To this end, we adapted a trap door biopsy method. Follicular density and quality of the biopsies were evaluated and ultrasound analysis was performed before and continuously after surgery to assess ovarian structure. Ovarian tissue biopsies recovered by the trap door technique allowed the successful harvesting of primordial follicles from capuchin monkeys, and no complication was recorded. The female cycle was not affected by surgery and no adherence was found thereafter. In conclusion, the adaptation of a trap door biopsy method is a safe procedure and allows recovery of healthy primordial follicles.
Assuntos
Folículo Ovariano/cirurgia , Procedimentos de Cirurgia Plástica/métodos , Animais , Biópsia , Cebus , Feminino , Humanos , Microscopia , Folículo Ovariano/citologia , Folículo Ovariano/diagnóstico por imagem , Ultrassonografia Doppler DuplaRESUMO
The efficiency of in vitro fertilization (IVF) depends on the viability of spermatozoa. For capuchin monkeys (Cebus apella), in vitro capacitation of spermatozoa is challenging because of their unique seminal coagulum. Motile spermatozoa can be obtained after liquefaction of the semen coagulum in coconut water-based solution. The objective of the present study was to establish an optimal in vitro maturation (IVM) protocol for capuchin monkeys and to observe the effect of follicle stimulating hormone (FSH) and luteinising hormone (LH) on IVF and parthenogenetic activation (PA) of oocytes collected from unstimulated females. We assessed spermatozoa quality after recovery from seminal coagulum using the solution ACP-118® as an extender. Oocytes were matured in vitro for 36 or 40 h and subjected to IVF or PA by applying ionomycin combined either with 6-dimethylaminopurine (6-DMAP) or roscovitine. In total, 87% of oocytes reached metaphase II (MII) after 40 IVM and 4-cell embryo production was obtained after IVF and parthenogenesis using ionomycin/6-DMAP. ACP-118® was used successfully to harvest viable spermatozoa from semen coagulum and in the preservation of spermatozoa, which were able to fertilize oocytes in vitro.
Assuntos
Embrião de Mamíferos/citologia , Fertilização in vitro , Metáfase/efeitos dos fármacos , Oócitos/citologia , Partenogênese/fisiologia , Espermatozoides/citologia , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Cebus , Células Cultivadas , Embrião de Mamíferos/efeitos dos fármacos , Feminino , Hormônio Foliculoestimulante/farmacologia , Hormônios/farmacologia , Hormônio Luteinizante/farmacologia , Masculino , Oócitos/efeitos dos fármacos , Partenogênese/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Purinas/farmacologia , Roscovitina , Espermatozoides/efeitos dos fármacosRESUMO
In the last decade, vitrification protocols to preserve human ovarian tissue have been regularly reported, even more often than the protocols developed for large mammals, such as ruminants and nonhuman primates. In order to facilitate the use of domestic ruminants (cows, goats, and sheep) and nonhuman primates as animal models, application of similar protocols as used for human material is performed. Next to it, the addition of indispensable or exclusion of avoidable compounds in the vitrification of human ovarian tissue should be tested in such experiments with animal models. The objective of this mini-review is to summarize the current protocols used for the vitrification of ovarian tissue and to evaluate the vitrification methods in humans, nonhuman primates, and domestic ruminants.
Assuntos
Criopreservação/métodos , Ovário/citologia , Vitrificação , Animais , Bancos de Espécimes Biológicos , Crioprotetores , Feminino , Humanos , Modelos Animais , Primatas , RuminantesRESUMO
The objectives of this study were to determine: 1) the optimal concentration (1.0 or 1.5 M) and duration of exposure (5, 10, or 20 min) of ovarian tissue to 1,2-propanediol (PROH) on morphology and viability of caprine preantral follicles; and 2) the effect of supplementing cryopreservation medium supplementation with Trolox(®) (0.1, 0.5, or 1.0 mM) or catalase (5, 10, or 20 IU/mL) on follicular morphology, viability, and lipid peroxidation. Cryopreservation decreased (p<0.05) percentages of normal follicles relative to the control (84%). Although supplementation of the cryopreservation medium (1.0 M PROH) with catalase (10 or 20 IU/mL) or Trolox(®) (0.1 mM) resulted in follicular morphology and viability similar to that in the controls (P>0.05), lipid peroxidation was reduced only when 20 IU/mL catalase was added to the cryopreservation medium.