983 resultados para Chick Embryo.
Resumo:
Seed coat is a specialized maternal tissue that interfaces the embryo and the external environment during embryogenesis, dormancy and germination. In addition, it is the first defensive barrier against penetration by pathogens and herbivores. Here we show that Albizia lebbeck seed coat dramatically compromises the oviposition, eclosion and development of the bruchid Callosobruchus maculatus. Dietary supplementation of bruchid larvae with A. lebbeck seed coat flour causes severe weight loss and reduces survival. By means of protein purification, mass spectrometry and bioinformatic analyses, we show that chitin-binding vicilins are the main source of A. lebbeck tegumental toxicity to C. maculatus. At concentrations as low as 0.1%, A. lebbeck vicilins reduce larval mass from 8.1 ± 1.7 (mass of control larvae) to 1.8 ± 0.5 mg, which corresponds to a decrease of 78%. Seed coat toxicity constitutes an efficient defense mechanism, hindering insect predation and preventing embryo damage. We hypothesize that A. lebbeck vicilins are good candidates for the genetic transformation of crop legumes to enhance resistance to bruchid predation.
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The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.
Resumo:
Preimplantation genetic diagnosis (PGD) was originally developed to diagnose embryo-related genetic abnormalities for couples who present a high risk of a specific inherited disorder. Because this technology involves embryo selection, the medical, bioethical, and legal implications of the technique have been debated, particularly when it is used to select features that are not related to serious diseases. Although several initiatives have attempted to achieve regulatory harmonization, the diversity of healthcare services available and the presence of cultural differences have hampered attempts to achieve this goal. Thus, in different countries, the provision of PGD and regulatory frameworks reflect the perceptions of scientific groups, legislators, and society regarding this technology. In Brazil, several texts have been analyzed by the National Congress to regulate the use of assisted reproduction technologies. Legislative debates, however, are not conclusive, and limited information has been published on how PGD is specifically regulated. The country requires the development of new regulatory standards to ensure adequate access to this technology and to guarantee its safe practice. This study examined official documents published on PGD regulation in Brazil and demonstrated how little direct oversight of PGD currently exists. It provides relevant information to encourage reflection on a particular regulation model in a Brazilian context, and should serve as part of the basis to enable further reform of the clinical practice of PGD in the country.
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Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract.
Resumo:
The function of the visceral yolk sac (VYS) is critical for embryo organogenesis until final fetal development in rats, and can be affected by conditions such as diabetes. In view of the importance of diabetes during pregnancy for maternal and neonatal health, the objective of this study was to assess fetal weight, VYS cell markers, and viability in female Wistar rats (200-250 g) with induced diabetes (alloxan, 37 mg/kg) on the 8th gestational day (gd 8). At gd 15, rats from control (n=5) and diabetic (n=5) groups were anesthetized and laparotomized to remove the uterine horns for weighing of fetuses and collecting the VYS. Flow cytometry was used for characterizing VYS cells, and for determining mitochondrial activity, cell proliferation, DNA ploidy, cell cycle phases, and caspase-3 activity. Fetal weight was reduced in the diabetic group. Expression of the cell markers CD34, VEGFR1, CD115, CD117, CD14, CCR2, CD90, CD44, STRO-1, OCT3/4, and Nanog was detected in VYS cells in both groups. In the diabetic group, significantly decreased expression of CD34 (P<0.05), CCR2 (P<0.001), and OCT3/4 (P<0.01), and significantly increased expression of CD90 (P<0.05), CD117 (P<0.01), and CD14 (P<0.05) were observed. VYS cells with inactive mitochondria, activated caspase-3, and low proliferation were present in the rats with diabetes. Severe hyperglycemia caused by maternal diabetes had negative effects on pregnancy, VYS cell viability, and the expression of cell markers.
Resumo:
PhotoAcoustic Imaging (PAI) is a branch in clinical and pre-clinical imaging, that refers to the techniques mapping acoustic signals caused by the absorption of the short laser pulse. This conversion of electromagnetic energy of the light to the mechanical (acoustic) energy is usually called photoacoustic effect. PAI, by combining optical excitation with acoustical detection, is able to preserve the diffraction limited spatial resolution. At the same time, the penetration depth is extended beyond the diffusive limit. The Laser-Scanning PhotoAcoustic Microscope system (LS-PAM) has been developed, that offers the axial resolution of 7.75 µm with the lateral resolution better than 10 µm. The first in vivo imaging experiments were carried out. Thus, in vivo label-free imaging of the mouse ear was performed. The principle possibility to image vessels located in deep layers of the mouse skin was shown. As well as that, a gold printing sample, vasculature of the Chick Chorioallantoic Membrane Assay, Drosophila larvae were imaged by PAI. During the experimental work, a totally new application of PAM was found, in which the acoustic waves, generated by incident light can be used for further imaging of another sample. In order to enhance the performance of the presented system two main recommendation can be offered. First, the current system should be transformed into reflection-mode setup system. Second, a more powerful source of light with the sufficient repetition rate should be introduced into the system.
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The objective of this study was to investigate the morphology, anatomy and germination behaviour of Phoenix roebelenii seeds. Biometric data were obtained by measuring 100 seeds extracted from recently harvested fruits and air-dried for one day. Four replications of 50 seeds each were previously treated with Vitavax-Thiran and then put to germinate in Sphagnum sp. in plastic trays at room temperature. Morphological details of the seeds were documented with the help of a scanning electronic microscope and then drawings were made with the help of a clear camera coupled to a stereomicroscope. Permanent lamina containing embryo sections were prepared to study its anatomy. The mean dimensions of the seeds were: length of 10.32mm, width of 5.21mm and thickness of 3.91mm. The weight of one thousand seeds was of 151.1g and the mean number of units.kg-1 was 6,600. Germination started between 27 and 58 days after sowing. The seeds are of the albuminous type, the endosperm is hard and the embryo (which is not clearly differentiated) occupies a lateral and peripheral position. During seed germination, seedling protrusion begins with the opening of an operculum, through which the cotyledon petiole is emitted with the embryonic axis at its tip. The portion of the cotyledon petiole that remains inside the seeds acts as a haustorium for the absorption of nutrients from the endosperm. The plumule emerges through a rift in the posterior part of the cotyledon. Secondary roots are observed to grow from the anterior part of the primary root.
Resumo:
Castor bean cropping has great social and economic value, but its production has been affected by factors such as low quality seeds used for sowing. The quick and precise evaluation of seed quality by x-ray test is known as an effective method to evaluate seed lots, but little is known about the interpretation between of the type of radiographic image and the seed quality correlation. The potential of x-ray analysis as a marker of seed physiological quality and as an initial process for the implementation of the use of computer-assisted image analysis was investigated using castor bean seeds of the different cultivars. The seeds were classified according to internal morphology visualized in the radiography and subjected to the germination test, emergency and seedling growth rate. It was possible to identify the different types of internal tissues, morphological and physical damage in castor bean seeds using the x-ray test. Tissues generating translucent images, embryo deformation, or tissues with less than 50% of endosperm reserves or spotted, negatively affected the physiological potential of the seed lots. Radiographic analysis is effective as an instrument to improve castor bean seed lot quality. This non destructive analysis allows the prediction of seedling performance and enabled the selection of high-quality seeds under the standards of a sustainable and precision agriculture
Resumo:
The decay of seeds is irreversible and at best can only be delayed by applying techniques that reduce the velocity of the metabolic reactions involved. There is little information on the biochemistry of tropical forest tree seeds related to their storability. It was investigated the influence of the composition of lipids and soluble sugars of two storage compartments, the cotyledons and the embryonic axis, of Melanoxylon brauna Schot. (Leguminosae- Caesalpinioideae), a hardwood known as black brauna, seeds stored at 20 ºC for 0, 3, 6, 9 and 12 months (natural ageing) and for 0, 24, 48, 72 and 96 hours at 40 ºC (accelerated ageing). The levels of fatty acids and monosaccharides varied differentially in each of the embryo storage compartments. Changes in oligosaccharide levels were similar for both types of ageing, diminishing in both compartments. Ageing can be attributed to the significant decrease of oligosaccharides and the increase of glucose in both types of ageing and both embryo compartments.
Resumo:
Freshly harvested triticale seeds are usually dormant, making the immediate evaluation of the physiological potential of seed lots difficult. We evaluated different triphenyl tetrazolium chloride (TTC) test methods for rapidly determining the viability of four seed lots of x.Triticosecale Wittmack cultivar IPR111. The test variables were: Preconditioning, (i) placing whole seeds between moistened paper towels or (ii) directly soaking the seeds in water, both procedures being conducted at 20 ºC for 18 hours; Post-conditioning seed preparation, (i) longitudinal bisection of the seed through the embryo with one half being stained and the other discarded or (ii) longitudinal bisection with both halves being stained; Staining for three and four hours, in the dark, with 0.1%, 0.5% or 1.0% (w/v) TTC according to the preconditioning method described above, (i) both halves of each seed were placed on filter paper moistened with TTC and maintained at 40 ºC or (ii) one half of each seed was immersed in 5 mL of TTC solution in a 100 mL glass beaker at 30 ºC. The best results were obtained by preconditioning seeds between moistened paper towels at 20 ºC for 18 hours and staining on filter paper with 1.0% (w/v) TTC for three hours at 40 ºC.
Resumo:
The tetrazolium test is used to control seed quality of various plant species since it allows a rapid evaluation of viability. Freshly harvested barley seeds show dormancy that can make the germination test ineffective for an immediate evaluation. Therefore, the development of more efficient methods, such as the tetrazolium test, is necessary. The objective of this research work was to study various procedures for performing the tetrazolium test on barley seeds. Five lots of cv. BRS 195 barley seeds were used and subjected to the following treatments: two different methods of seed preconditioning (direct immersion in H2O and between sheets of moistened paper towels); two types of preparation for staining (longitudinal cross-section of the seed through the embryo with immersion of one half in a 2,3,5 triphenyl tetrazolium chloride solution or placing both halves on top of filter paper moistened with the tetrazolium salt solution); two methods of staining (on top of filter paper and direct immersion in the tetrazolium salt solution). Three concentrations of the tetrazolium salt solution (0.1%, 0.5%, and 1.0%) were used. It was concluded that the tetrazolium test on barley seeds may be accomplished with preconditioning by direct immersion in H2O and staining by immersing in a 0.1% or 0.5% concentration of tetrazolium salt solution or staining on top of filter paper moistened with such solution at a 1.0% concentration.
Resumo:
Xylopia aromatica is a native species from Brazil's "Cerrado", recommended for restoration ecology and also as a medicine. Its seeds have embryos with morphophysiological dormancy, making nursery propagation difficult. The objective of this study was to verify the efficiency of X-ray and tetrazolium tests for evaluating the viability of three seed lots, stored for different periods. All seeds were X-rayed (13 kV, 350 seconds) and samples used for tetrazolium and germination tests. In the tetrazolium test, seeds were submitted to six treatments at two temperatures (25 and 30 °C) with imbibition in distilled water and immersion in three concentrations of tetrazolium solution (0.5, 0.75 and 1%) at the two imbibition temperatures. Seeds for the germination test were placed for imbibition in distilled water and a 500 ppm Promalin® (6-Benzyladenine + GA4 + GA7) solution and later sown in sterilized sand. The embryo could not be observed with the X-ray test. However, those seeds observed with an undamaged endosperm did not differ in the percentages of seeds with firm and stained endosperms observed in the tetrazolium test for all the lots. The tetrazolium test is efficient for evaluating seed viability, principally if imbibed at 30 °C and immersed in a 0.5% solution at 30 °C.
Resumo:
Structural differences such as abnormalities, damage and free spaces in seeds may affect germination. The aim of this study was to study the relationship between eggplant seed morphology and seed germination. Ten seed lots of the eggplant cultivar Embu were evaluated by X-ray image analysis and the germination test. Seed image analysis was performed by Image Pro Plus® software and the whole seed area and free space between the embryo and endosperm were measured. The internal seed area filled by the embryo and endosperm was calculated from the difference between the whole seed and free space areas. Based on these results and visual seed analysis, seeds were classified into three categories and information on germination was obtained for each one. X-ray image analysis provides a perfect view of the internal seed parts and for seed morphology studies. An increase in seed area filled by the endosperm and embryo does not improve seed germination. Mechanical seed damage and deteriorated tissues can adversely affect seed germination.
Resumo:
One component of successful parenting is related to efficiency in foraging behaviour. The relationships among chick feeding, the size and type of food package, and length of parental foraging trips has not been well studied in seabirds. In addition, relatively few data have been collected on the activities of seabirds when foraging away from the nest site. The objectives of this study were: (1) to contrast productivity, feeding rate, and attendance patterns of individuals carrying a novel transmitter with a control group of birds; (2) to use radio-telemetry to assess the variability in foraging locations within and between individual male Common Terns; (3) to determine the seasonal variation in chick diet; (4) to determine for each transmittered bird, the relationships among the foraging patterns, parental behaviour, and seasonal reproductive success. The study took place over two years (1990-91) on a concrete breakwater 1 km offshore on Lake Erie near Port Colbome, Ontario. Ten pairs of terns in 1990 and 12 pairs in 1991 were radio-tracked by boat or car during the chick rearing stage. Concurrent behavioural observations documented the time each sex spent foraging or at the nest. The frequency and prey species composition of feeds to chicks were also recorded. The transmitters had negligible effects on the feeding frequency and brood attendance patterns of transmitter carrying birds. Peak nesting transmittered birds in 1990 and 1991 exhibited some inter-individual variability in foraging locations, however intraindividual variability was low. Birds foraged primarily to the west and northwest of the colony. Late nesters exhibited greater inter-individual variability, however intra-individual variability remained low for most birds. Neither group demonstrated sufficient variability to support the regular use of this colony as an "information centre". Individual transmittered birds had unique and predictable foraging patterns, and corresponding differences in feeding frequencies and brood attendance patterns, yet productivity was essentially equal between nests due to the impact and importance of stochastic events. Individuals that were recaptured in 1991 exhibited very similar foraging patterns to 1990, suggesting littie variability between years. Conservation of foraging patterns between years may have potential implications for mate choice decisions in future breeding seasons. Prey species delivered to chicks differed between morning and evening for peak and late nesters in 1990, but not 1991. Peak nesters in 1990 fed significantiy more Rainbow Smelt fOsmerus mordM) than Emerald Shiner (Notropis atherinoidesV this trend was reversed for late nesters who also fed large numbers of unidentified larval fish. No significant differences were found in 1991. Seasonal changes in prey species delivered to chicks is believed to be attributable to the temperature tolerances of the smelt and shiners, and the presence of large schools of larval fish during the late nesting season.
Resumo:
Increased losses of eggs and chicks resulting from human intrusion (investigator or other) into seabird colonies has been well documented. In 1990/91, I studied the effects of investigator disturbance on aggressive behaviour and breeding success of individual pairs of ring-billed gulls nesting at two colonies near Port Colborne, Ontario. The insular colony was on an artificial breakwall, associated with the Welland Ship Canal, approximately 1 km off the north shore of Lake Erie. The mainland colony was adjacent to the canal approximately 1 km east of the breakwall. The frequencies of adult threat and assault behaviours, chick movement and adult attacks on chicks were recorded by continuous scan sampling 30 min prior to, 30 min during and 60 (2 X 30) min after investigator disturbance. The frequency of threat and assault behaviours increased during the period of investigator activity in the colony while the duration of wingpulls and beakpulls decreased. Significantly more chicks ran ("runners") from their natal territories during disturbances and "runners" were more frequently attacked than "territorial" chicks. No chicks were fatally attacked during disturbance and "runners" returned to their natal territories quickly after disturbance. Breeding success was determined for pairs nesting in study plots subjected to two levels of disturbance (normal and moderate). The disturbance level of each plot differed in visitation frequency and activities performed on each visit. Investigator disturbance had no effect on the hatching success or fledging success (taken as 21 days of age) of ring-billed gull study pairs at either colony.