ABSTRACT
While cryopreservation of cauda epididymal sperm (SpCau) allows the preservation of post-mortem bulls' gametes, the process triggers sperm damage. Although improving post-thaw sperm quality, using egg yolk extenders (EY) raises biosafety concerns which forces the use of EY-free extenders (EYFE). Since EYFE are less efficient in preserving post-thaw sperm quality, a strategy for ejaculated sperm (SpEj) frozen with EYFE is to add an Equilibrium Time (ET) step period to the cryopreservation process. However, the ET effect on the quality of SpCau cryopreserved in EYFE remains unknown. Distinct from SpEJ, SpCau physiologically displays cytoplasmic droplets (CDs) in the flagellum that may benefit cell exchange during ET. We hypothesized that using ET in SpCau cryopreserved with EYFE impacts sperm morphofunctional features, CD area, and in vitro fertility ability. Extender nanoparticles were also assessed. Following collection from the cauda epididymis of six Nellore bulls by retrograde flow, SpCau were cryopreserved in EYFE BoviFree® (Minitube, Germany) using three ET protocols: ET0 (no-ET); ET2.5 (2.5 h-ET); and ET5 (5 h-ET). SpCau from ET2.5 and ET5 showed a higher (P ≤ 0.05) percentage of motility and integrity of plasma and acrosome membranes and a smaller (P ≤ 0.05) distal CD area. There are no differences in sperm abnormalities, oxidative stress, capacitation-like events, and in vitro fertility ability. However, a better sperm recovery was found after Percoll® selection for ET2.5 and ET5. Interestingly, the number of nanoparticles in the extender decreased in post-thawed samples. In conclusion, an ET of 2.5 or 5 h is required for an efficient SpCau cryopreservation using an EYFE.
Subject(s)
Cryopreservation , Cryoprotective Agents , Epididymis , Nanoparticles , Semen Preservation , Sperm Motility , Spermatozoa , Male , Animals , Cryopreservation/methods , Cryopreservation/veterinary , Semen Preservation/methods , Semen Preservation/veterinary , Cryoprotective Agents/pharmacology , Spermatozoa/cytology , Epididymis/cytology , Cattle , Nanoparticles/chemistry , Egg Yolk/chemistry , Semen Analysis , CytoplasmABSTRACT
This study investigated the seasonal variations in mRNA expression of FSH (Fshr), LH (Lhr) receptors, melatonin (Mt1 and Mt2) receptors, melatonin-synthetizing enzymes (Asmt and Aanat) and melatonin concentration in developing follicles from mares raised in natural photoperiods. For one year, ultrasonographic follicular aspiration procedures were performed monthly, and small (<20 mm), medium (20 to 35 mm) and large (>35 mm) follicles were recovered from five mares. One day before monthly sample collections, an exploratory ultrasonography conducted to record the number and the size of all follicles larger than 15 mm. The total number of large follicles were higher during the spring/summer (8.2 ± 1.9) than during autumn/winter (3.0 ± 0.5). Compared to autumn/winter seasons, there was an increase of Fshr and Aanat mRNA expressions in small, medium and large follicles, an increase of Lhr and Asmt mRNA expressions in medium and large follicles and an increase of Mt1 and Mt2 mRNA expressions in small and large follicles during spring/summer. The melatonin levels in follicular fluid were also higher during the spring/summer seasons. The present data show that melatonin locally upregulates the mRNA expression of Mt1 and Mt2 receptors and melatonin-forming enzymes in mare developing follicles during reproductive seasons.
ABSTRACT
Early embryonic development occurs in the oviduct, where an ideal microenvironment is provided by the epithelial cells and by the oviductal fluid produced by these cells. The oviductal fluid contains small extracellular vesicles (sEVs), which through their contents, including microRNAs (miRNAs), can ensure proper cell communication between the mother and the embryo. However, little is known about the modulation of miRNAs within oviductal epithelial cells (OECs) and sEVs from the oviductal fluid in pregnant cows. In this study, we evaluate the miRNAs profile in sEVs from the oviductal flushing (OF-sEVs) and OECs from pregnant cows compared to non-pregnant, at 120 h after ovulation induction. In OF-sEVs, eight miRNAs (bta-miR-126-5p, bta-miR-129, bta-miR-140, bta-miR-188, bta-miR-219, bta-miR-345-3p, bta-miR-4523, and bta-miR-760-3p) were up-regulated in pregnant and one miRNA (bta-miR-331-5p) was up-regulated in non-pregnant cows. In OECs, six miRNAs (bta-miR-133b, bta-miR-205, bta-miR-584, bta-miR-551a, bta-miR-1193, and bta-miR-1225-3p) were up-regulated in non-pregnant and none was up-regulated in pregnant cows. Our results suggest that embryonic maternal communication mediated by sEVs initiates in the oviduct, and the passage of gametes and the embryo presence modulate miRNAs contents of sEVs and OECs. Furthermore, we demonstrated the transcriptional levels modulation of selected genes in OECs in pregnant cows. Therefore, the embryonic-maternal crosstalk potentially begins during early embryonic development in the oviduct through the modulation of miRNAs in OECs and sEVs in pregnant cows.
ABSTRACT
In freshwater fish with external fertilization, sperm sampling can be contaminated with urine, which triggers motility and gives rise to decreased fertilization success. The maintenance of freshwater fish in hyperosmotic conditions may reduce urine production and improve sperm quality. Thus, the aim of this work was to verify if acute exposure to various NaCl concentrations improves sperm quality in the yellowtail tetra Astyanax altiparanae. Spermiation was induced using a single dose of carp pituitary gland (5 mg kg-1) and the males were maintained at various NaCl concentrations: NaCl 0.00% (control), NaCl 0.45% (hypoosmotic), NaCl 0.9% (isosmotic) and NaCl 1.0% (hyperosmotic) for 6 h at 26 °C. Sperm was collected and verified for activation by urine and motility traits. At 0.00%, 0.45%, and 0.90%, the sperm was motile just after sampling, indicating activation by urine. Surprisingly, at hyperosmotic conditions, no activation was observed. Other sperm and motility parameters did not show any statistical differences, including sperm viability (P = 0.7083), concentration (P = 0.9030), total motility (P = 0.6149), VCL (curvilinear velocity; P = 0.1216), VAP (average path velocity; P = 0.1231) and VSL (straight-line velocity; P = 0.1340). Our results indicate that acute maintenance at hyperosmotic conditions eliminates sperm activation by urine and maintains sperm quality. Such a new procedure is interesting for both basic and applied sciences, including reproductive practice in fish.(AU)
Em peixes de água doce com fertilização externa, a amostragem de espermatozoides pode ser contaminada pela urina, o que desencadeia motilidade e gera menor sucesso na fertilização. A manutenção de peixes de água doce em condições hiperosmóticas pode reduzir a produção de urina e melhorar a qualidade do esperma. Assim, o presente trabalho foi delineado para verificar se a exposição aguda a várias concentrações de NaCl melhora a qualidade do esperma no tetra-amarelo Astyanax altiparanae. A espermiação foi induzida usando uma dose única de hipófise da carpa (5 mg kg-1) e os machos foram mantidos em várias concentrações de NaCl: NaCl 0,00% (controle), NaCl 0,45% (hipoosmótico), NaCl 0,9% (isosmótico) e NaCl 1,0% (hiperosmótico) por seis horas a 26 °C. O esperma foi colhido e verificado quanto à ativação por urina e traços de motilidade. Em 0,00%, 0,45%, 0,90% os espermatozóides eram móveis logo após a amostragem, indicando ativação pela urina. Surpreendentemente, em condições hiperosmóticas, nenhuma ativação foi observada. Outros parâmetros espermáticos e de motilidade não mostraram diferenças estatísticas, incluindo viabilidade espermática (P = 0,7083), concentração (P = 0,9030), motilidade total (P = 0,6149), VCL (Velocidade Curvilinear; P = 0,1216), VMD (Velocidade Média de Deslocamento; P = 0,1230) e VLR (Velocidade em linha Reta; P = 0,1340). Nossos resultados indicam que a manutenção aguda em condições hiperosmóticas elimina a ativação do esperma pela urina e mantém a qualidade do esperma. Esse novo procedimento é interessante para as ciências básicas e aplicadas, incluindo a prática reprodutiva em peixes.(AU)
Subject(s)
Animals , Osmosis , Salinity , Semen Analysis/methods , Semen Analysis/veterinary , Characidae/physiology , Sperm MotilityABSTRACT
In freshwater fish with external fertilization, sperm sampling can be contaminated with urine, which triggers motility and gives rise to decreased fertilization success. The maintenance of freshwater fish in hyperosmotic conditions may reduce urine production and improve sperm quality. Thus, the aim of this work was to verify if acute exposure to various NaCl concentrations improves sperm quality in the yellowtail tetra Astyanax altiparanae. Spermiation was induced using a single dose of carp pituitary gland (5 mg kg-1) and the males were maintained at various NaCl concentrations: NaCl 0.00% (control), NaCl 0.45% (hypoosmotic), NaCl 0.9% (isosmotic) and NaCl 1.0% (hyperosmotic) for 6 h at 26 °C. Sperm was collected and verified for activation by urine and motility traits. At 0.00%, 0.45%, and 0.90%, the sperm was motile just after sampling, indicating activation by urine. Surprisingly, at hyperosmotic conditions, no activation was observed. Other sperm and motility parameters did not show any statistical differences, including sperm viability (P = 0.7083), concentration (P = 0.9030), total motility (P = 0.6149), VCL (curvilinear velocity; P = 0.1216), VAP (average path velocity; P = 0.1231) and VSL (straight-line velocity; P = 0.1340). Our results indicate that acute maintenance at hyperosmotic conditions eliminates sperm activation by urine and maintains sperm quality. Such a
Em peixes de água doce com fertilização externa, a amostragem de espermatozoides pode ser contaminada pela urina, o que desencadeia motilidade e gera menor sucesso na fertilização. A manutenção de peixes de água doce em condições hiperosmóticas pode reduzir a produção de urina e melhorar a qualidade do esperma. Assim, o presente trabalho foi delineado para verificar se a exposição aguda a várias concentrações de NaCl melhora a qualidade do esperma no tetra-amarelo Astyanax altiparanae. A espermiação foi induzida usando uma dose única de hipófise da carpa (5 mg kg-1) e os machos foram mantidos em várias concentrações de NaCl: NaCl 0,00% (controle), NaCl 0,45% (hipoosmótico), NaCl 0,9% (isosmótico) e NaCl 1,0% (hiperosmótico) por seis horas a 26 °C. O esperma foi colhido e verificado quanto à ativação por urina e traços de motilidade. Em 0,00%, 0,45%, 0,90% os espermatozóides eram móveis logo após a amostragem, indicando ativação pela urina. Surpreendentemente, em condições hiperosmóticas, nenhuma ativação foi observada. Outros parâmetros espermáticos e de motilidade não mostraram diferenças estatísticas, incluindo viabilidade espermática (P = 0,7083), concentração (P = 0,9030), motilidade total (P = 0,6149), VCL (Velocidade Curvilinear; P = 0,1216), VMD (Velocidade Média de Deslocamento; P = 0,1230) e VLR (Velocidade em linha Reta; P = 0,1340). Nossos resultados indicam que a m
ABSTRACT
In freshwater fish with external fertilization, sperm sampling can be contaminated with urine, which triggers motility and gives rise to decreased fertilization success. The maintenance of freshwater fish in hyperosmotic conditions may reduce urine production and improve sperm quality. Thus, the aim of this work was to verify if acute exposure to various NaCl concentrations improves sperm quality in the yellowtail tetra Astyanax altiparanae. Spermiation was induced using a single dose of carp pituitary gland (5 mg kg-1) and the males were maintained at various NaCl concentrations: NaCl 0.00% (control), NaCl 0.45% (hypoosmotic), NaCl 0.9% (isosmotic) and NaCl 1.0% (hyperosmotic) for 6 h at 26 °C. Sperm was collected and verified for activation by urine and motility traits. At 0.00%, 0.45%, and 0.90%, the sperm was motile just after sampling, indicating activation by urine. Surprisingly, at hyperosmotic conditions, no activation was observed. Other sperm and motility parameters did not show any statistical differences, including sperm viability (P = 0.7083), concentration (P = 0.9030), total motility (P = 0.6149), VCL (curvilinear velocity; P = 0.1216), VAP (average path velocity; P = 0.1231) and VSL (straight-line velocity; P = 0.1340). Our results indicate that acute maintenance at hyperosmotic conditions eliminates sperm activation by urine and maintains sperm quality. Such a
Em peixes de água doce com fertilização externa, a amostragem de espermatozoides pode ser contaminada pela urina, o que desencadeia motilidade e gera menor sucesso na fertilização. A manutenção de peixes de água doce em condições hiperosmóticas pode reduzir a produção de urina e melhorar a qualidade do esperma. Assim, o presente trabalho foi delineado para verificar se a exposição aguda a várias concentrações de NaCl melhora a qualidade do esperma no tetra-amarelo Astyanax altiparanae. A espermiação foi induzida usando uma dose única de hipófise da carpa (5 mg kg-1) e os machos foram mantidos em várias concentrações de NaCl: NaCl 0,00% (controle), NaCl 0,45% (hipoosmótico), NaCl 0,9% (isosmótico) e NaCl 1,0% (hiperosmótico) por seis horas a 26 °C. O esperma foi colhido e verificado quanto à ativação por urina e traços de motilidade. Em 0,00%, 0,45%, 0,90% os espermatozóides eram móveis logo após a amostragem, indicando ativação pela urina. Surpreendentemente, em condições hiperosmóticas, nenhuma ativação foi observada. Outros parâmetros espermáticos e de motilidade não mostraram diferenças estatísticas, incluindo viabilidade espermática (P = 0,7083), concentração (P = 0,9030), motilidade total (P = 0,6149), VCL (Velocidade Curvilinear; P = 0,1216), VMD (Velocidade Média de Deslocamento; P = 0,1230) e VLR (Velocidade em linha Reta; P = 0,1340). Nossos resultados indicam que a m
ABSTRACT
In freshwater fish with external fertilization, sperm sampling can be contaminated with urine, which triggers motility and gives rise to decreased fertilization success. The maintenance of freshwater fish in hyperosmotic conditions may reduce urine production and improve sperm quality. Thus, the aim of this work was to verify if acute exposure to various NaCl concentrations improves sperm quality in the yellowtail tetra Astyanax altiparanae. Spermiation was induced using a single dose of carp pituitary gland (5 mg kg-1) and the males were maintained at various NaCl concentrations: NaCl 0.00% (control), NaCl 0.45% (hypoosmotic), NaCl 0.9% (isosmotic) and NaCl 1.0% (hyperosmotic) for 6 h at 26 °C. Sperm was collected and verified for activation by urine and motility traits. At 0.00%, 0.45%, and 0.90%, the sperm was motile just after sampling, indicating activation by urine. Surprisingly, at hyperosmotic conditions, no activation was observed. Other sperm and motility parameters did not show any statistical differences, including sperm viability (P = 0.7083), concentration (P = 0.9030), total motility (P = 0.6149), VCL (curvilinear velocity; P = 0.1216), VAP (average path velocity; P = 0.1231) and VSL (straight-line velocity; P = 0.1340). Our results indicate that acute maintenance at hyperosmotic conditions eliminates sperm activation by urine and maintains sperm quality. Such a new procedure is interesting for both basic and applied sciences, including reproductive practice in fish.(AU)
Em peixes de água doce com fertilização externa, a amostragem de espermatozoides pode ser contaminada pela urina, o que desencadeia motilidade e gera menor sucesso na fertilização. A manutenção de peixes de água doce em condições hiperosmóticas pode reduzir a produção de urina e melhorar a qualidade do esperma. Assim, o presente trabalho foi delineado para verificar se a exposição aguda a várias concentrações de NaCl melhora a qualidade do esperma no tetra-amarelo Astyanax altiparanae. A espermiação foi induzida usando uma dose única de hipófise da carpa (5 mg kg-1) e os machos foram mantidos em várias concentrações de NaCl: NaCl 0,00% (controle), NaCl 0,45% (hipoosmótico), NaCl 0,9% (isosmótico) e NaCl 1,0% (hiperosmótico) por seis horas a 26 °C. O esperma foi colhido e verificado quanto à ativação por urina e traços de motilidade. Em 0,00%, 0,45%, 0,90% os espermatozóides eram móveis logo após a amostragem, indicando ativação pela urina. Surpreendentemente, em condições hiperosmóticas, nenhuma ativação foi observada. Outros parâmetros espermáticos e de motilidade não mostraram diferenças estatísticas, incluindo viabilidade espermática (P = 0,7083), concentração (P = 0,9030), motilidade total (P = 0,6149), VCL (Velocidade Curvilinear; P = 0,1216), VMD (Velocidade Média de Deslocamento; P = 0,1230) e VLR (Velocidade em linha Reta; P = 0,1340). Nossos resultados indicam que a manutenção aguda em condições hiperosmóticas elimina a ativação do esperma pela urina e mantém a qualidade do esperma. Esse novo procedimento é interessante para as ciências básicas e aplicadas, incluindo a prática reprodutiva em peixes.(AU)
Subject(s)
Animals , Osmosis , Salinity , Semen Analysis/methods , Semen Analysis/veterinary , Characidae/physiology , Sperm MotilityABSTRACT
Establishment of pregnancy after embryo transfer is the ultimate goal of an embryo transfer program and increasing pregnancy rates and reducing pregnancy loss are mandatory. The utilization of treatments to improve conception rates in recipient mares has been the focus of several research groups over the last years and the results are controversial. Some studies using human chorionic gonadotrophin (hCG) found promising results. Our hypothesis was that hCG administration would cause an additional stimulation on luteal function, uterine and luteal vascularization and progesterone concentration, and the mares would have increased uterine and cervix tone. Therefore, in the present study the effects of hCG administration to induce ovulation, on day 0 (day of ovulation) or day 5 postovulation were evaluated on corpus luteum characteristics, reproductive tract vascularization, and serum progesterone concentration from ovulation until day 15 postovulation. Groups were: G1: (control) - no hCG; G2: 2500 IU of hCG to induce ovulation when a follicle greater than 35mm and uterine edema were detected; G3: 2500 IU hCG on day 0; G4: 2500 IU hCG on day 5 postovulation. Twelve mares were randomly assigned to each group, during consecutive cycles, in a Latin Square experimental design, in a total of 48 cycles. Doppler ultrasound evaluations were performed daily from day 0 until day 15 postovulation, including mesometrial vascularity, endometrial vascularity and corpus luteum vascularity. Blood samples were collected for serum progesterone concentration. Data was analyzed using the Proc Glimmix SAS Procedure for nonparametric variables and Proc Mixed for parametric parameters. There was no treatment effect for all variables studied (P > 0.05). Characteristics were only affected by day (P < 0.05). It can be concluded that hCG administration at the time points suggested in the current study did not alter the characteristics evaluated.
ABSTRACT
The aim was to evaluate the associations between circulating P4 concentrations, corpus luteum (CL) size (diameter, area or volume) and blood perfusion (BP) in cows. In Experiment 1, Pearson's correlations (P < 0.05) with P4 concentrations were observed during CL development (D8) for total area (TA; r = 0.76), luteal area (ACL; r = 0.72), total and luteal diameter (TD and DCL respectively; r = 0.46). During mid-late diestrus, there was a positive correlation (P < 0.05) only at D15 with TA and ACL (r > 0.60), TD, total volume (TV) and luteal volume (VCL; r > 0.434). During luteal regression, the correlation was only observed at D18 for ACL (r = 0.478) and D20 with several variables. In Experiment 2, CL weight and ACL had the greatest correlation with P4 (r > 0.6). In Experiment 3, TA and ACL were the variables that were most closely correlated with serum P4 concentrations at D7 in recipient cows. Correlation coefficients were greater for luteal measurements when there were compact compared with cavitary CLs. In Experiment 4, there was no correlation (P > 0.05) between P4 and any of the variables measured on D4 and D7 in recipient cows detected in estrus. On D18 to D20, all CL characteristics were correlated (P < 0.05) with plasma P4, and luteal BP and BP area were more closely (P < 0.05) correlated than ACL. In conclusion, CL perimeter area measurements had the greatest association with luteal function during CL development; whereas for BP there was a greater correlation with P4 than luteal size during luteolysis.
Subject(s)
Cattle/physiology , Corpus Luteum/anatomy & histology , Estrous Cycle/physiology , Pregnancy, Animal , Progesterone/blood , Ultrasonography/veterinary , Animals , Corpus Luteum/physiology , Female , Ovulation , Pregnancy , Pregnancy, Animal/physiologyABSTRACT
Establishment of pregnancy after embryo transfer is the ultimate goal of an embryo transfer program and increasing pregnancy rates and reducing pregnancy loss are mandatory. The utilization of treatments to improve conception rates in recipient mares has been the focus of several research groups over the last years and the results are controversial. Some studies using human chorionic gonadotrophin (hCG) found promising results. Our hypothesis was that hCG administration would cause an additional stimulation on luteal function, uterine and luteal vascularization and progesterone concentration, and the mares would have increased uterine and cervix tone. Therefore, in the present study the effects of hCG administration to induce ovulation, on day 0 (day of ovulation) or day 5 postovulation were evaluated on corpus luteum characteristics, reproductive tract vascularization, and serum progesterone concentration from ovulation until day 15 postovulation. Groups were: G1: (control) - no hCG; G2: 2500 IU of hCG to induce ovulation when a follicle greater than 35mm and uterine edema were detected; G3: 2500 IU hCG on day 0; G4: 2500 IU hCG on day 5 postovulation. Twelve mares were randomly assigned to each group, during consecutive cycles, in a Latin Square experimental design, in a total of 48 cycles. Doppler ultrasound evaluations were performed daily from day 0 until day 15 postovulation, including mesometrial vascularity, endometrial vascularity and corpus luteum vascularity. Blood samples were collected for serum progesterone concentration. Data was analyzed using the Proc Glimmix SAS Procedure for nonparametric variables and Proc Mixed for parametric parameters. There was no treatment effect for all variables studied (P > 0.05). Characteristics were only affected by day (P < 0.05). It can be concluded that hCG administration at the time points suggested in the current study did not alter the characteristics evaluated.
Subject(s)
Female , Animals , Cattle , Biomarkers/analysis , Horses/physiology , Estrous Cycle/physiology , Chorionic Gonadotropin/analysis , Embryo Transfer/veterinary , Corpus Luteum/physiologyABSTRACT
Establishment of pregnancy after embryo transfer is the ultimate goal of an embryo transfer program and increasing pregnancy rates and reducing pregnancy loss are mandatory. The utilization of treatments to improve conception rates in recipient mares has been the focus of several research groups over the last years and the results are controversial. Some studies using human chorionic gonadotrophin (hCG) found promising results. Our hypothesis was that hCG administration would cause an additional stimulation on luteal function, uterine and luteal vascularization and progesterone concentration, and the mares would have increased uterine and cervix tone. Therefore, in the present study the effects of hCG administration to induce ovulation, on day 0 (day of ovulation) or day 5 postovulation were evaluated on corpus luteum characteristics, reproductive tract vascularization, and serum progesterone concentration from ovulation until day 15 postovulation. Groups were: G1: (control) - no hCG; G2: 2500 IU of hCG to induce ovulation when a follicle greater than 35mm and uterine edema were detected; G3: 2500 IU hCG on day 0; G4: 2500 IU hCG on day 5 postovulation. Twelve mares were randomly assigned to each group, during consecutive cycles, in a Latin Square experimental design, in a total of 48 cycles. Doppler ultrasound evaluations were performed daily from day 0 until day 15 postovulation, including mesometrial vascularity, endometrial vascularity and corpus luteum vascularity. Blood samples were collected for serum progesterone concentration. Data was analyzed using the Proc Glimmix SAS Procedure for nonparametric variables and Proc Mixed for parametric parameters. There was no treatment effect for all variables studied (P > 0.05). Characteristics were only affected by day (P < 0.05). It can be concluded that hCG administration at the time points suggested in the current study did not alter the characteristics evaluated.(AU)
Subject(s)
Animals , Female , Cattle , Chorionic Gonadotropin/analysis , Estrous Cycle/physiology , Biomarkers/analysis , Horses/physiology , Embryo Transfer/veterinary , Corpus Luteum/physiologyABSTRACT
In fish with external fertilization, sperm must reach the oocyte through the micropyle to enter the cytoplasm. Fertilization success is then influenced by characteristics of oocytes or sperm. In this study, we evaluated oocyte morphology and sperm motility parameters and their effects on the inseminating dose in a teleost fish Astyanax altiparanae. Interestingly, we found one of the lowest yet described inseminating doses in teleosts (2390 spermatozoa oocyte-1 ml-1). Such a fertilization efficacy may be explained by the long duration of sperm motility (>75 s), the small oocyte diameter (695.119 µm), large micropyle diameter (7.57 µm), and the presence of grooves on the oocyte surface that guides spermatozoon to the fertilization area. Additionally, we have described for the first time a structure that combines grooves on the chorion surface and a ridge in the micropylar area.
Subject(s)
Fertilization in Vitro , Fishes/physiology , Ovum/physiology , Sperm-Ovum Interactions/physiology , Spermatozoa/physiology , Animals , Female , Male , Ovum/cytology , Spermatozoa/cytologyABSTRACT
Oocytes that undergo in vitro maturation (IVM) are metabolically abnormal and accumulate excess lipid content. However, the mechanism of lipid accumulation and the role of cumulus cells in this process are unclear. Recently, it was shown that fatty acid binding proteins (FABPs) performed intra- and extracellular fatty acid transport. We postulated that FABP3 might be responsible for fatty acid transport from cumulus cells to the oocytes via transzonal projections (TZPs) during IVM. Transcript and protein levels of FABP3 were analyzed in both in vivo- and in vitro-matured cumulus-oocyte-complexes and were increased in IVM samples. Further analysis showed increased lipid content in oocytes and cumulus cells in IVM samples compared to in vivo-derived. We therefore speculated that altered traffic of fatty acids via FABP3 during IVM was the mechanism leading to the excess of lipids accumulated within IVM oocytes. Furthermore, we demonstrated an increase in FABP3 levels and lipid content during the first 9 h of IVM, further strengthening the possibility of fatty acid transport via FABP3 and TZPs. Additionally, disruptions of TZPs during IVM decreased lipid accumulation in oocytes. Our results shed light on a possible mechanism involving FABP3 and TZPs that causes excess lipid accumulation in oocytes during IVM.
Subject(s)
Cumulus Cells/metabolism , Fatty Acid Binding Protein 3/metabolism , In Vitro Oocyte Maturation Techniques/methods , Lipid Metabolism , Oocytes/metabolism , Animals , Cattle , In Vitro TechniquesABSTRACT
Follicular development and deviation processes during the postpartum period are not completely known in horses. Thus, we aimed to study the characteristics of follicular dynamics and ovarian vascular perfusion during the postpartum period in mares that demonstrated estrous behavior and had early (<10 days) or late (≥10 days) postpartum ovulation. Ten mares were scanned daily by transrectal ultrasonography from the first day postpartum (d1) to the sixteenth day after the first postpartum ovulation (D0 = ovulation). The animals were split in the early (n = 3) and late (n = 7) ovulation groups (average interval between parturition and ovulation: 8.0 ± 0.0 and 14.7 ± 1.2 days, respectively). For the follicular growth, no difference (P > 0.1) was detected between the groups when the data were normalized for the days preceding the first postpartum ovulation (from D-7 to D-1). However, when the data were normalized to days postpartum, the dominant follicle was larger (P < 0.05) in the early-ovulated group in all days during this period (d1 to d7). The number of follicles >25 mm diameter was greater (P < 0.05) in the early-ovulated group during the first 3 days postpartum, and the late-ovulated mares showed greater number of follicles with 20-25 mm during d4 to d7. For blood flow characteristics, no difference (P > 0.1) was detected between groups in vascular perfusion of the dominant follicle or in the ovarian pedicle ipsilateral to the largest follicle. Similarly, no difference (P > 0.1) was detected in progesterone concentrations, corpus luteum (CL) area and vascular perfusion of the CL. Pregnancy rate did not differ (P > 0.1) between the early (3/3; 100%) and late (5/7; 71.4%) groups. Therefore, the characteristics of the follicle growth on the preceding days of ovulation were similar between the early- and late-ovulated mares and were consistent with the follicular dynamics expected in non-pregnant and non-lactating mares. However, when the data were analyzed for the days relative to parturition, greater follicle development was present in mares that ovulate earlier during the postpartum period (<10 days). The results suggest that important events may occur before parturition, resulting in early follicle development, mainly in those mares that show estrus and ovulate within 10 days postpartum.
Subject(s)
Horses/physiology , Ovary/blood supply , Ovary/physiology , Ovulation/physiology , Postpartum Period , Animals , FemaleABSTRACT
Tambaqui is the major native Characiform fish species in Brazil. The loin-eye area (LEA) is a feature currently used in breeding programs for livestock and may be a phenotype susceptible to selection also in tambaquis. The purpose of this study was to evaluate the efficiency of two different LEA measurements. LEA data were taken using the vegetable paper (VP) and ultrasonography (US) techniques measured in three LEA regions (LEA1) at the end of the operculum region, (LEA2) at the insertion of the dorsal fin, and (LEA3) in the region of insertion of the anal fin from 29 individuals in a fish processing plant located in the State of Rondônia, Brazil. The data were analyzed in the R program using both the Students t-test and the Pearson correlation. The mean values for the two methods were not statistically different at the 5% risk level, indicating that the US can efficiently estimate the LEA compared with the actual value given by VP. The correlation was moderate (0.61) for LEA2 and can be attributed to the loin cut into two halves lengthwise in tambaquis usually done during commercial fish processing. Ultrasound has proven to be an efficient method for accurately estimating the LEA in tambaqui for further genetic breeding programs.(AU)
Tambaqui é a maior espécie de peixe da ordem Characiformes no Brasil. A área de olho de lombo (AOL) é uma característica usada atualmente em programas de melhoramento genético em animais de criação terrestre. O objetivo deste estudo foi avaliar a eficiência de dois tipos diferentes de medidas de AOL. As medidas de AOL foram tomadas com o uso de papel vegetal (PV) e ultrassonografia (US) em três regiões do animal: (AOL1) região final da região do opérculo, (AOL2) na inserção da nadadeira dorsal e (AOL3) na região da inserção da nadadeira caudal em 29 indivíduos advindos de uma planta processadora localizada no estado de Rondônia, Brasil. Os dados foram analisados no programa R com o uso do test t de Student e a correlação de Pearson. Os valores médios para ambas metodologias não foram estatisticamente diferentes ao nível de 5%, indicando que US pode eficientemente estimar a AOL quando comparado com o valor real gerado por PV. A correlação foi moderada (0,61) para AOL2 e isto pode ser atribuído ao corte em duas metades do lombo geralmente realizado em tambaquis nas plantas de processamento. A técnica de ultrassom provou ser um método eficiente para estimar acuradamente a AOL em tambaqui em futuros programas de melhoramento genético.(AU)
Subject(s)
Animals , Characiformes/anatomy & histology , Body Weights and Measures/veterinary , Animal Fins/anatomy & histology , Animal Fins/diagnostic imaging , Ultrasonography/veterinaryABSTRACT
Tambaqui is the major native Characiform fish species in Brazil. The loin-eye area (LEA) is a feature currently used in breeding programs for livestock and may be a phenotype susceptible to selection also in tambaquis. The purpose of this study was to evaluate the efficiency of two different LEA measurements. LEA data were taken using the vegetable paper (VP) and ultrasonography (US) techniques measured in three LEA regions (LEA1) at the end of the operculum region, (LEA2) at the insertion of the dorsal fin, and (LEA3) in the region of insertion of the anal fin from 29 individuals in a fish processing plant located in the State of Rondônia, Brazil. The data were analyzed in the R program using both the Students t-test and the Pearson correlation. The mean values for the two methods were not statistically different at the 5% risk level, indicating that the US can efficiently estimate the LEA compared with the actual value given by VP. The correlation was moderate (0.61) for LEA2 and can be attributed to the loin cut into two halves lengthwise in tambaquis usually done during commercial fish processing. Ultrasound has proven to be an efficient method for accurately estimating the LEA in tambaqui for further genetic breeding programs.
Tambaqui é a maior espécie de peixe da ordem Characiformes no Brasil. A área de olho de lombo (AOL) é uma característica usada atualmente em programas de melhoramento genético em animais de criação terrestre. O objetivo deste estudo foi avaliar a eficiência de dois tipos diferentes de medidas de AOL. As medidas de AOL foram tomadas com o uso de papel vegetal (PV) e ultrassonografia (US) em três regiões do animal: (AOL1) região final da região do opérculo, (AOL2) na inserção da nadadeira dorsal e (AOL3) na região da inserção da nadadeira caudal em 29 indivíduos advindos de uma planta processadora localizada no estado de Rondônia, Brasil. Os dados foram analisados no programa R com o uso do test t de Student e a correlação de Pearson. Os valores médios para ambas metodologias não foram estatisticamente diferentes ao nível de 5%, indicando que US pode eficientemente estimar a AOL quando comparado com o valor real gerado por PV. A correlação foi moderada (0,61) para AOL2 e isto pode ser atribuído ao corte em duas metades do lombo geralmente realizado em tambaquis nas plantas de processamento. A técnica de ultrassom provou ser um método eficiente para estimar acuradamente a AOL em tambaqui em futuros programas de melhoramento genético.
Subject(s)
Animals , Characiformes/anatomy & histology , Animal Fins/anatomy & histology , Animal Fins/diagnostic imaging , Body Weights and Measures/veterinary , Ultrasonography/veterinaryABSTRACT
Tambaqui is the major native Characiform fish species in Brazil. The loin-eye area (LEA) is a feature currently used in breeding programs for livestock and may be a phenotype susceptible to selection also in tambaquis. The purpose of this study was to evaluate the efficiency of two different LEA measurements. LEA data were taken using the vegetable paper (VP) and ultrasonography (US) techniques applied in three LEA regions: (LEA1) at the end of the operculum region, (LEA2) at the insertion of the dorsal fin, and (LEA3) in the region of insertion of the anal fin from 29 individuals in a fish processing plant located in the State of Rondônia, Brazil. The data were analyzed in the R program using both the Students t-test and the Pearson correlation. The mean values for the two methods were not statistically different at the 5% risk level, indicating that the US can efficiently estimate the LEA compared with the actual value given by VP. The correlation was moderate (0.61) for LEA2 and can be attributed to the loin cut into two halves lengthwise in tambaquis, usually done during commercial fish processing. Ultrasound has proven to be an efficient method for accurately estimating the LEA in tambaqui for further genetic breeding programs.
Tambaqui é a maior espécie de peixe da ordem Characiformes no Brasil. A área de olho de lombo (AOL) é uma característica usada atualmente em programas de melhoramento genético em animais de criação terrestre. O objetivo deste estudo foi avaliar a eficiência de dois tipos diferentes de medidas de AOL. As medidas de AOL foram tomadas com o uso de papel vegetal (PV) e ultrassonografia (US) em três regiões do animal: (AOL1), região final da região do opérculo, (AOL2), na inserção da nadadeira dorsal, e (AOL3), na região da inserção da nadadeira caudal, em 29 indivíduos advindos de uma planta processadora localizada no estado de Rondônia, Brasil. Os dados foram analisados no programa R com o uso do test-t de Student e a correlação de Pearson. Os valores médios para ambas as metodologias não foram estatisticamente diferentes em nível de 5%, indicando que US pode eficientemente estimar a AOL quando comparada com o valor real gerado por PV. A correlação foi moderada (0,61) para AOL2 e isto pode ser atribuído ao corte em duas metades do lombo geralmente realizado em tambaquis nas plantas de processamento. A técnica de ultrassom provou ser um método eficiente para estimar acuradamente a AOL em tambaquis em futuros programas de melhoramento genético.
ABSTRACT
Fluorescent probes (propidium iodide, Hoechst 33342, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin, and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide) were used to simultaneously evaluate the integrity of plasma and acrosomal membranes as well as mitochondrial membrane function in cryopreserved bovine semen and to verify its influence on fertility and postinsemination uterine vascularization. One hundred eighty-two Nellore cows were distributed for artificial insemination (AI) using semen batches separated according to the cell percentage presenting intact plasma membrane, intact acrosome, and high mitochondrial function (IPIAH): group G (44.5% IPIAH, n = 68), group M (23.0% IPIAH, n = 56), and group R (8.5% IPIAH, n = 58). The uterine hemodynamic was evaluated by Doppler sonogram in three periods: 30 hours before AI, 4 and 24 hours after AI were considered the resistance index and the uterine vascularization score. The pregnancy rate of group G (64.7%) was greater (P > 0.05) compared with group R (36.2%), but both did not differ from group M (50.0%). There was no effect (P > 0.05) of semen quality on uterine vascularization. Greater vascularization was noticed 4 hours after AI than 30 hours before and 24 hours after AI. Semen evaluation using fluorescent probes contributes to predicting fertilizing potential of semen. The use of semen with less percentage of IPIAH sperm does not alter uterine hemodynamic in cows.
Subject(s)
Cattle/physiology , Fertility/physiology , Fluorescent Dyes , Insemination, Artificial/veterinary , Semen Analysis/veterinary , Uterus/blood supply , Animals , Female , Hemodynamics/physiology , Insemination, Artificial/methods , Male , Pregnancy , Pregnancy Rate , Semen Analysis/methodsABSTRACT
Introduction: Efficient laboratory services are the basis of modern health systems. Scientific innovations have contributed to substantial improvements in the laboratory environment, but errors still persist. These errors are classified as pre-analytical, analytical and post-analytical, according to the time of occurrence. Objective: To evaluate the frequency of pre-analytical errors in the clinical laboratory service of a military hospital. Methods: A total of 329,582 tests were performed in the clinical laboratory of Hospital Naval Marcílio Dias (HNMD) from August to October 2012, and pre-analytical errors were documented. Results: The most frequent cause of the observed pre-analytical errors was hemolysis (27.54%), followed by samples not received (25.43%) and insufficient sample volume (18.49%). The samples from the Integrated Home Care Service (SIAD) showed the highest frequency of errors (3.38%), followed by those from the inpatient (0.76%) and the outpatient departments (0.21%). Conclusion: Our study demonstrates the importance of managing laboratory pre-analytical quality in order to ensure service excellence...
Introdução: Serviços laboratoriais efetivos são a base dos sistemas de saúde modernos. Inovações científicas têm contribuído para melhorias substanciais no meio laboratorial, mas os erros ainda persistem. Esses erros são classificados como pré-analíticos, analíticos e pós-analíticos, dependendo do momento de sua ocorrência. Objetivo: Avaliar a frequência de erros pré-analíticos ocorridos no serviço de análises clínicas de um hospital militar. Métodos: Um total de 329.582 exames foram realizados no laboratório de análises clínicas do Hospital Naval Marcílio Dias no período de agosto a outubro de 2012, e os erros pré-analíticos documentados. Resultados: Os erros pré-analíticos mais observados foram decorrentes da hemólise (27,54%), seguidos de material não recebido (25,43%) e amostra insuficiente (18,49%). As amostras oriundas do Serviço Integrado de Atendimento Domiciliar (SIAD) foram as que apresentaram a maior frequência de erros (3,38%), seguidas pelo setor de pacientes internos (0,76%) e ambulatoriais (0,21%), respectivamente. Conclusão: Nosso estudo demonstra a importância da gestão da fase pré-analítica na garantia da qualidade laboratorial, de maneira a assegurar um serviço de excelência...
Subject(s)
Humans , Total Quality Management , Indicators of Health Services , Laboratories, Hospital/standards , Diagnostic Errors/statistics & numerical data , Laboratories, Hospital/organization & administration , Quality ControlABSTRACT
In spite of numerous, substantial advances in equine reproduction, many stages of embryonic and fetal morphological development are poorly understood, with no apparent single source of comprehensive information. Hence, the objective of the present study was to provide a complete macroscopic and microscopic description of the equine embryo/fetus at various gestational ages. Thirty-four embryos/fetuses were aged based on their crown rump length (CRL), and submitted to macroscopic description, biometry, light and scanning microscopy, as well as the alizarin technique. All observed developmental changes were chronologically ordered and described. As examples of the main observed features, an accentuated cervical curvature was observed upon macroscopic examination in all specimens. In the nervous system, the encephalic fourth ventricle and the encephalic vesicles forebrain, midbrain, and hindbrain, were visualized from Day 19 (ovulation = Day 0). The thoracic and pelvic limbs were also visualized; their extremities gave rise to the hoof during development from Day 27. Development of other structures such as pigmented optical vesicle, liver, tail, cardiac area, lungs, and dermal vascularization started on Days 25, 25, 19, 19, 34, and 35, respectively. Light and scanning microscopy facilitated detailed examinations of several organs, e.g., heart, kidneys, lungs, and intestine, whereas the alizarin technique enabled visualization of ossification. Observations in this study contributed to the knowledge regarding equine embryogenesis, and included much detailed data from many specimens collected over a long developmental interval.