Energetic Costs of Stress in Developing Fishes: Quantifying Allostasis and Allostatic Load.

Stress exerts negative effects on fish health through stimulation of the hypothalamic-pituitary-interrenal axis and autonomic nervous system, resulting in heightened neural and neuroendocrine responses. Energetic investment and physiological adaptation are then required to re-establish homeostatic stability or reach a new allostatic state. The cost of the energetic investment is referred to as allostatic load (AL). While determining the sources of stress and assessing their consequences have resulted in estimates of AL, most of this work has been conducted in adult mammals and humans; no ALs exist for developing fish. From a series of experiments on a model species, zebrafish (Danio rerio), whose yolk-sac larvae were exposed to two chronic stressors (high-temperature and hypoxia), ALs were quantified based on biomarkers of ontogenetic changes in growth, morphometrics, and metabolic activities. Results showed that for zebrafish yolk-sac larvae, chronic stress imposed high AL and, thus, high total allostatic energetic costs, (Rt (AL)), because of prolonged energy demand in the face of limited resources (e.g., yolk). Under severe chronic stress, energetic costs were sufficiently large that energy-limited developing fish may not be able to fully compensate, resulting in maladaptive responses from allostatic overload, leading either to death or to novel allostatic states, possibly more resilient to environmental change.

Expression of genes associated with apoptosis in the residual yolk sac during the perihatch period of broiler chicks.

The yolk sac (YS) consists of the yolk and the surrounding YS tissue, which provides essential nutrients and physiological functions for the developing embryo. After the YS is internalized into the abdominal cavity of the embryonic chick, the YS starts to degrade. Apoptosis, or programmed cell-death, is speculated to be the mechanism behind degradation of the YS. The objective of this study was to determine if degradation of the YS tissue was mediated by apoptosis during the perihatch period. The YS tissue was collected from broiler chicks from embryonic d 17 to d 7 posthatch. The mRNA abundance of genes that are involved in the regulation, initiation, and execution of apoptosis were analyzed by qPCR. The mRNA for Bcl2, Bcl2L11, cytochrome C and caspases 3, 6, 7, 8, 9, and 18 all showed a linear response from embryonic d 17 to d 7 posthatch. To confirm the role of apoptosis in the degradation of the YS tissue, a DNA fragmentation assay was performed. Degradation of genomic DNA in the YS tissue started on day of hatch. The characteristic ladder of oligonucleosomal-sized DNA fragments was observed on d 3, 5, and 7 posthatch. The combined gene expression and DNA fragmentation results demonstrate that degradation of the YS posthatch is mediated by apoptosis.

A common epigenetic mechanism across different cellular origins underlies systemic immune dysregulation in an idiopathic autism mouse model.

Immune dysregulation plays a key role in the pathogenesis of autism. Changes occurring at the systemic level, from brain inflammation to disturbed innate/adaptive immune in the periphery, are frequently observed in patients with autism; however, the intrinsic mechanisms behind them remain elusive. We hypothesize a common etiology may lie in progenitors of different types underlying widespread immune dysregulation. By single-cell RNA sequencing (sc-RNA seq), we trace the developmental origins of immune dysregulation in a mouse model of idiopathic autism. It is found that both in aorta-gonad-mesonephros (AGM) and yolk sac (YS) progenitors, the dysregulation of HDAC1-mediated epigenetic machinery alters definitive hematopoiesis during embryogenesis and downregulates the expression of the AP-1 complex for microglia development. Subsequently, these changes result in the dysregulation of the immune system, leading to gut dysbiosis and hyperactive microglia in the brain. We further confirm that dysregulated immune profiles are associated with specific microbiota composition, which may serve as a biomarker to identify autism of immune-dysregulated subtypes. Our findings elucidate a shared mechanism for the origin of immune dysregulation from the brain to the gut in autism and provide new insight to dissecting the heterogeneity of autism, as well as the therapeutic potential of targeting immune-dysregulated autism subtypes.

Multiple cell populations generate macrophage progenitors in the early yolk sac.

Yolk sac (YS) CSF1 receptor positive (CSF1R+) cells are thought to be the progenitors for tissue-resident macrophages present in various tissues. The YS progenitors for tissue-resident macrophages are referred to as erythroid-myeloid progenitors (EMPs). However, diverse types of hematopoietic progenitors are present in the early YS, thus it is not precisely known which type of hematopoietic cell gives rise to the CSF1R+ lineage. In this study, an analysis was conducted to determine when CSF1R+ progenitors appeared in the early YS. It showed that CSF1R+ cells appeared in the YS as early as embryonic day 9 (E9) and that the earliest hematopoietic progenitors that differentiate into CSF1R+ cells were found in E8. Since these progenitors possessed the capability to generate primitive erythroid cells, it was likely that primitive erythroid lineages shared progenitors with the CSF1R+ lineage. Mutual antagonism appears to work between PU.1 and GATA1 when CSF1R+ cells appear in the early YS. One day later (E9), multiple progenitors, including myeloid-restricted progenitors and multipotent progenitors, in the YS could immediately generate CSF1R+ cells. These results suggest that EMPs are not an exclusive source for the CSF1R+ lineage; rather, multiple hematopoietic cell populations give rise to CSF1R+ lineage in the early YS.

Interaction of extraembryonic microglia and neonatal brain development.

Microglia are resident immune cells in the central nervous system (CNS), which, in a healthy state, promote CNS homeostasis and respond to CNS injury. In contrast, microglia are also implicated in pathological conditions where they may contribute to neural damage. Primitive microglia arise from extraembryonic progenitors in the yolk sac (YS). The extraembryonic origins of primitive microglia are distinct from other tissue macrophages. The YS is the first site of hematopoiesis in development. Uniquely, microglial pregenital cells in the mouse derive from an early myeloid branch of the hematopoietic lineage in the YS. Microglia are critical in several key stages of physiological brain development, including embryonic vasculogenesis, immunosurveillance, and neurogenesis. Abnormal microglial function has been linked to neurodevelopmental and neurodegenerative diseases, although mechanistic roles in disease etiology remain incompletely understood. Knowledge of species-specific differences between human, murine and other animal models is also critical to understanding translational relevance to human health and disease as biomedical understanding of the importance of primitive microglia advances. This significance drives the importance of understanding, comparatively, the extraembryonic origins and developmental mechanisms whereby human primitive microglia differentiate and migrate to inform translational research. A better understanding of the molecular drivers may lead to biomarkers and/or preventative or therapeutic measures for neonatal brain development and neurodegenerative diseases. Herein, the role of microglia in neonatal brain development is discussed, current understandings of the developmental origins of microglia are described, the ontogeny and phylogeny of microglia, and implications of in vitro microglia-like cell differentiation, with a specific interest on neurodegenerative diseases, are reviewed. Together, these emphasize the importance of leveraging the extraembryonic origins of microglia to not only better understand neurodevelopment and neurodegenerative diseases, but also to develop protective measures that are specific to human microglia.

Placentation in the Human and Higher Primates.

Placentation in humans is precocious and highly invasive compared to other mammals. Implantation is interstitial, with the conceptus becoming completely embedded within the endometrium towards the end of the second week post-fertilization. Villi initially form over the entire surface of the chorionic sac, stimulated by histotrophic secretions from the endometrial glands. The secondary yolk sac never makes contact with the chorion, and a choriovitelline placenta is never established. However, recent morphological and transcriptomic analyses suggest that the yolk sac plays an important role in the uptake of nutrients from the coelomic fluid. Measurements performed in vivo demonstrate that early development takes place in a physiological, low-oxygen environment that protects against teratogenic free radicals and maintains stem cells in a multipotent state. The maternal arterial circulation to the placenta is only fully established around 10-12 weeks of gestation. By then, villi have regressed over the superficial, abembryonic pole, leaving the definitive discoid placenta, which is of the villous, hemochorial type. Remodeling of the maternal spiral arteries is essential to ensure a high-volume but low-velocity inflow into the mature placenta. Extravillous trophoblast cells migrate from anchoring villi and surround the arteries. Their interactions with maternal immune cells release cytokines and proteases that are key to remodeling, and a successful pregnancy.

The road (not) taken - Placental transfer and interspecies differences.

Species differences are among the main reasons for the high failure rate of preclinical studies. A better awareness and understanding of these differences might help to improve the outcome of preclinical research. In reproduction, the placenta is the central organ regulating fetal exposure to a substance circulating in the maternal organism. Exact information about placental transfer can help to better estimate the toxic potential of a substance. From an evolutionary point of view, the chorioallantoic placenta is the organ with the highest anatomical diversity among species. Moreover, frequently used animal models in reproduction belong to rodents and lagomorphs, two groups that are characterized by the generation of an additional type of placenta, which is crucial for fetal development, but absent from humans: the inverted yolk sac placenta. Taken together, the translatability of placental transfer studies from laboratory animals to humans is challenging, which is supported by the fact that numerous species-dependent toxic effects are described in literature. Thus, reliable human-relevant data are frequently lacking and the toxic potential of chemicals and pharmaceuticals for humans can hardly be estimated, often resulting in recommendations that medical treatments or exposure to chemicals should be avoided for safety reasons. Although species differences of placental anatomy have been described frequently and the need for human-relevant research models has been emphasized, analyses of substances with species-dependent placental transfer have been performed only sporadically. Here, we present examples for species-specific placental transfer, including that of nanoparticles and pharmaceuticals, and discuss potential underlying mechanisms. With respect to the COVID 19-pandemic it might be of interest that some antiviral drugs are reported to feature species-specific placental transfer. Further, differences in placental structure and antibody transfer may affect placental transfer of ZIKA virus.

Yolk sac, but not hematopoietic stem cell-derived progenitors, sustain erythropoiesis throughout murine embryonic life.

In the embryo, the first hematopoietic cells derive from the yolk sac and are thought to be rapidly replaced by the progeny of hematopoietic stem cells. We used three lineage-tracing mouse models to show that, contrary to what was previously assumed, hematopoietic stem cells do not contribute significantly to erythrocyte production up until birth. Lineage tracing of yolk sac erythromyeloid progenitors, which generate tissue resident macrophages, identified highly proliferative erythroid progenitors that rapidly differentiate after intra-embryonic injection, persisting as the major contributors to the embryonic erythroid compartment. We show that erythrocyte progenitors of yolk sac origin require 10-fold lower concentrations of erythropoietin than their hematopoietic stem cell-derived counterparts for efficient erythrocyte production. We propose that, in a low erythropoietin environment in the fetal liver, yolk sac-derived erythrocyte progenitors efficiently outcompete hematopoietic stem cell progeny, which fails to generate megakaryocyte and erythrocyte progenitors.

Centennial Review: The chicken yolk sac is a multifunctional organ.

The yolk sac (YS) consists of the yolk, which supplies nutrients, and the YS tissue, which surrounds the yolk and provides essential metabolic functions for the developing embryo. The YS tissue is derived from the midgut of the embryo and consists of a layer of endodermal epithelial cells (EEC) in contact with the yolk contents, a mesodermal layer that contains the vascular system and an outer ectodermal layer. The YS tissue is a multifunctional organ that provides essential functions such as host immunity, nutrient uptake, carbohydrate and lipid metabolism, and erythropoiesis. The YS tissue plays a role in immunity by the transport of maternal antibodies in the yolk to the embryonic circulation that feeds the developing embryo. In addition, the YS tissue expresses high mRNA levels of the host defense peptide, avian ß-defensin 10 during mid embryogenesis. Owing to its origin, the YS EEC share some functional properties with intestinal epithelial cells such as expression of transporters for amino acids, peptides, monosaccharides, fatty acids, and minerals. The YS tissue stores glycogen and expresses enzymes for glycogen synthesis and breakdown and glucogenesis. This carbohydrate metabolism may play an important role in the hatching process. The mesodermal layer of the YS tissue is the site for erythropoiesis and provides erythrocytes before the maturation of the bone marrow. Other functions of the YS tissue involve synthesis of plasma proteins, lipid transport and cholesterol metabolism, and synthesis of thyroxine. Thus, the YS is an essential organ for the growth, development, and health of the developing embryo. This review will provide an overview of the studies that have investigated the functionalities of the YS tissue at the cellular and molecular levels with a focus on chickens.

Effect of posthatch feed and water access time on residual yolk and broiler live performance.

This study investigated the effect of feed and water access time on yolk sac utilization and subsequent broiler live performance. Hatching eggs were collected from commercial flocks of Ross 308 breeders at 35 and 39 wk of age in experiments 1 and 2, respectively. Chicks already out of their shells that still had some dampness on their down were removed, recorded, feather-sexed, and weighed at 488 h of incubation in both experiments. Chicks were weighed individually and received feed and water at 2 (immediate feed; IF), 8, 12, 16, 20, 24, 28, and 32 h after hatching (488 h) in experiments 1 and 2 (IF) and at 24, 26, 28, 32, 36, and 40 h after hatching in experiment 2. The residual yolk sac weight was determined at 32 and 40 h after hatching (day 0) in all groups in experiments 1 and 2, respectively. Feed consumption and BW were recorded at 7, 14, 21, and 35 d and at the same age relative to placement on feed and water at the end of the growing period. Mortality was recorded twice daily in both experiments. Feed and water access time did not influence yolk sac utilization in either experiment (P > 0.05). The IF group exhibited a higher (P < 0.05) BW than those that received feed at or after 28 h at 35 d in both experiments. There was a significant increase in feed consumption in the IF group compared with the groups with access to feed and water after 24 h at 35 d in experiment 2 (P < 0.05), with a similar trend in experiment 1 (P > 0.05). There were no significant differences in the feed conversion ratio (FCR) or mortality at 35 d of age, but the IF group tended to have a poorer FCR than the other groups in both experiments. When the total feed and water times were equalized among all groups, irrespective of the deprivation duration, there were no significant differences among the groups in the BW, feed consumption, the FCR, or mortality in both experiments. It can be concluded that feed and water deprivation for 28 h or longer after hatching (≥28 h) negatively affects the final BW but tends to improve the FCR at 35 d of age compared with chicks that receive feed immediately (2 h after hatching). When the feeding period was equalized in all groups, feed and water deprivation up to 40 h under optimum conditions had no detrimental effect on final live performance. These results suggest that the total feeding period is more critical for broiler performance than the time of posthatch access to feed and water.

Yolk sac hematopoiesis: does it contribute to the adult hematopoietic system?

In most vertebrates, the yolk sac (YS) represents the very first tissue where blood cells are detected. Therefore, it was thought for a long time that it generated all the blood cells present in the embryo. This model was challenged using different animal models, and we now know that YS hematopoietic precursors are mostly transient although their contribution to the adult system cannot be excluded. In this review, we aim at properly define the different waves of blood progenitors that are produced by the YS and address the fate of each of them. Indeed, in the last decade, many evidences have emphasized the role of the YS in the emergence of several myeloid tissue-resident adult subsets. We will focus on the development of microglia, the resident macrophages in the central nervous system, and try to untangle the recent controversy about their origin.

A review on yolk sac utilization in poultry.

During incubation, embryonic growth and development are dependent on nutrients deposited in the egg. The content of the yolk can be transferred to the embryo in 2 ways: directly into the intestine via the yolk stalk or through the highly vascularized yolk sac membrane. It has been suggested that, as a result of genetic selection and improved management, the increase in posthatch growth rate and concurrently the increase in metabolic rate of broiler chickens during the last 50 yr has also increased embryonic metabolism. A higher metabolic rate during incubation would imply a lower residual yolk weight and possibly lower energy reserve for the hatchling. This might affect posthatch development and performance. This review examined scientific publications published between 1930 and 2018 to compare residual yolk weight at hatch, metabolic heat production, and yolk utilization throughout incubation. This review aimed to investigate 1) whether or not residual yolk weight and composition has been changed during the 88-yr period considered and 2) which abiotic and biotic factors affect yolk utilization in poultry during incubation and the early posthatch period. It can be concluded that 1) residual yolk weight and the total solid amount of the residual yolk at hatch seem to be decreased in the recent decades. It cannot be concluded whether the (lack of) differences between old and modern strains are due to genetic selection, changed management and incubation conditions, or moment of sampling (immediately after hatch or at pulling). It is remarkable that with the genetic progress and improved management and incubation conditions over the last 88 yr, effects on yolk utilization efficiency and embryonic metabolic heat production are limited; 2) factors specially affecting residual yolk weight at hatch include egg size and incubation temperature, whereas breeder age has more influence on nutrient composition of the residual yolk.

In ovo injection of branched-chain amino acids: Embryonic development, hatchability and hatching quality of turkey poults.

In this study, the influence of a branched-chain amino acid blend (BCAA composed of 3 l-leucine:1 l-valine:2 l-isoleucine) injected into the amniotic fluid was evaluated for embryonic growth, yolk-sac (YS) utilization and development of gastrointestinal tract (GIT) and skeletal muscles of turkey embryos from day 24 of incubation (24E) to hatching, together with hatchability, poult quality and liver L* (lightness), a* (redness) and b* (yellowness) values at hatch. At day 22 of incubation, embryonated eggs (n = 240) were assigned to three treatments, that is, eggs were not injected (control, NC) or injected with 1.5 ml sterile solution with 0.9% salt (SA) or 0.2% BCAA blend (BCAAb). These solutions were injected manually into the amniotic fluid of the embryonated eggs. To determine weights and lengths (where appropriate) of the studied organs and tissues, four embryonated eggs and poults per treatment were selected at 24E and at hatch. While the BCAAb decreased the YS and embryo weight, hatchability and the liver L* value, it increased the weight and quality of poults and the weights of breast and thigh muscles at hatch. In conclusion, the in ovo feeding of the BCAA blend negatively affected hatchability but positively affected hatching weight and poult quality by improving development of skeletal muscles and by regulating energy metabolism.

Effect of chick body temperature during post-hatch handling on broiler live performance.

The current study investigated the effect of chick body (rectal) temperature during the post-hatch handling period on body weight (BW) loss, yolk sac utilization, organ weights, and broiler live performance. Hatching eggs were obtained from a commercial flock of Ross 308 broiler breeders at 44 wk of age. A total of 384 chicks were separated into 3 groups during the 12 h post-hatch handling period: control, high and low temperature groups, with average body temperatures of 40.0, 42.6, and 38.1°C, respectively. Residual yolk sac weight was not affected by temperature group, whereas the weights of organs such as the heart, gizzard, proventriculus, and bursa of Fabricius were significantly lower in the high body temperature group than in the control and low body temperature groups. BW was significantly lower at placement in chicks in the high temperature group than in chicks in the control and low body temperature groups due to greater weight loss during the post-hatch handling period (P ≤ 0.05). Lower BW was maintained in the chicks in the high body temperature group than in the chicks in the other 2 groups until the end of the experiment at 35 d (P ≤ 0.05) because chicks in the high temperature group consumed less feed throughout the experiment (P ≤ 0.05). Feed conversion ratio (FCR) and mortality were numerically greater in the high body temperature chicks than in the control group, whereas FCR and the mortality in the low body temperature chicks were intermediate at 35 d. The results of the present study indicate that day-old chicks with high body temperatures (42.6°C) exhibited a greater percentage of BW loss due to dehydration and lower organ weights during the 12 h post-hatch handling period, which was followed by significantly poorer broiler performance. There were no significant differences in performance between the chicks in the control (40.0°C) and low (38.1°C) body temperature groups. In conclusion, day-old chicks are more sensitive to higher body temperatures than to lower temperatures during the post-hatch handling period.

Sox17 is essential for proper formation of the marginal zone of extraembryonic endoderm adjacent to a developing mouse placental disk.

In mouse conceptus, two yolk-sac membranes, the parietal endoderm (PE) and visceral endoderm (VE), are involved in protecting and nourishing early-somite-stage embryos prior to the establishment of placental circulation. Both PE and VE membranes are tightly anchored to the marginal edge of the developing placental disk, in which the extraembryonic endoderm (marginal zone endoderm: ME) shows the typical flat epithelial morphology intermediate between those of PE and VE in vivo. However, the molecular characteristics and functions of the ME in mouse placentation remain unclear. Here, we show that SOX17, not SOX7, is continuously expressed in the ME cells, whereas both SOX17 and SOX7 are coexpressed in PE cells, by at least 10.5 days postconception. The Sox17-null conceptus, but not the Sox7-null one, showed the ectopic appearance of squamous VE-like epithelial cells in the presumptive ME region, together with reduced cell density and aberrant morphology of PE cells. Such aberrant ME formation in the Sox17-null extraembryonic endoderm was not rescued by the chimeric embryo replaced with the wild-type gut endoderm by the injection of wild-type ES cells into the Sox17-null blastocyst, suggesting the cell autonomous defects in the extraembryonic endoderm of Sox17-null concepti. These findings provide direct evidence of the crucial roles of SOX17 in proper formation and maintenance of the ME region, highlighting a novel entry point to understand the in vivo VE-to-PE transition in the marginal edge of developing placenta.

Omega-3 polyunsaturated fatty acids promote angiogenesis in placenta derived mesenchymal stromal cells.

Enhancement of angiogenesis is solicited in wound repair and regeneration. Mesenchymal stromal cells derived from the placenta (P-MSCs) have an inherent angiogenic potential. Polyunsaturated fatty acids (PUFAs) in turn, specifically the omega-3 (N-3) are essential for growth and development. They are also recommended as dietary supplements during pregnancy. We therefore hypothesized that addition of N-3 PUFAs in P-MSC culture media may enhance their angiogenic potential. Hence, we treated P-MSCs with omega-3 (N-3) fatty acids -Docosahexaenoic acid (DHA) and Eicosapentaenoic acid (EPA) at different concentrations and tested their angiogenic potential. We saw an upregulation of both bFGF and VEGFA. We also found enhanced in vitro tube formation ability of P-MSCs treated with DHA: EPA. We then looked at the influence of the conditioned medium (CM) collected from P-MSCs exposed to DHA: EPA on the key effector cells -HUVECs (Human Umbilical Vein derived endothelial cells and their functionality was further confirmed on chick yolk sac membrane. We found that the CM of P-MSCs exposed to DHA: EPA could enhance angiogenesis in both cases. These result were finally validated in an in vivo matrigel plug assay which revealed enhanced migration and vessel formation in CM treated with DHA: EPA. Our data thus reveals for the first time that supplementation with lower concentration of PUFA enhances the angiogenic potential of P-MSCs making them suitable for chronic wound healing applications.

Classics revisited: Miguel Fernández on germ layer inversion and specific polyembryony in armadillos.

BACKGROUND: Miguel Fernández was an Argentinian zoologist who published the first account of obligate polyembryony in armadillos. His contribution is here discussed in relation to his contemporaries, Newman and Patterson, and more recent work. FINDINGS: Fernandez worked on the mulita (Dasypus hybridus). He was able to get early stages before twinning occurred and show it was preceded by inversion of the germ layers. By the primitive streak stage there were separate embryonic shields and partition of the amnion. There was, however, a single exocoelom and all embryos were enclosed in a common set of membranes comprising chorion towards the attachment site in the uterine fundus and inverted yolk sac on the opposite face. He showed that monozygotic twinning did not occur in another armadillo, the peludo (Chaetophractus villosus). CONCLUSIONS: Fernández's work represented a major breakthrough in understanding how twinning occurred in armadillos. His work and that of others is of intrinsic interest to zoologists and has a direct bearing on the origin of monozygotic twins and birth defects in humans.

The effects of fluctuating temperature regimes on the embryonic development of lake whitefish (Coregonus clupeaformis).

Fluctuating incubation temperatures may have significant effects on fish embryogenesis; yet most laboratory-based studies use constant temperatures. For species that experience large, natural seasonal temperature changes during embryogenesis, such as lake whitefish (Coregonus clupeaformis), seasonal temperature regimes are likely optimal for development. Anthropogenic activities can increase average and/or variability of natural incubation temperatures over large (e.g. through climate change) or smaller (e.g. thermal effluent discharge) geographic scales. To investigate this, we incubated lake whitefish embryos under constant (2, 5, or 8°C) and fluctuating temperature regimes. Fluctuating temperature regimes had a base temperature of 2°C with: 1) seasonal temperature changes that modeled natural declines/inclines; 2) tri-weekly +3°C, 1h temperature spikes; or 3) both seasonal temperature changes and temperature spikes. We compared mortality to hatch, morphometrics, and heart rate at three developmental stages. Mortality rate was similar for embryos incubated at constant 2°C, constant 5°C, or with seasonal temperatures, but was significantly greater at constant 8°C. Embryos incubated constantly at >2°C had reduced body growth and yolk consumption compared to embryos incubated with seasonal temperature changes. When measured at the common base temperature of 2°C, embryos incubated at constant 2°C had lower heart rates than embryos incubated with both seasonal temperature changes and temperature spikes. Our study suggests that incubating lake whitefish embryos with constant temperatures may significantly alter development, growth, and heart rate compared to incubating with seasonal temperature changes, emphasizing the need to include seasonal temperature changes in laboratory-based studies.

Comparison of hatching egg characteristics, embryo development, yolk absorption, hatch window, and hatchability of Pekin Duck eggs of different weights.

This study was carried out to determine the hatching egg characteristics, embryo development and yolk absorption during incubation, hatch window, and hatchability of Pekin duck eggs of different weights. A total of 960 hatching eggs was obtained from a breeder flock 35 to 36 wk of age. The eggs were classed into 3 weight categories: "light" (L; <75 g), "medium" (M; 76 to 82 g), and "heavy" (H; >83 g). The albumen weight was the highest in the heavy eggs, whereas the yolk weight was higher in the medium and heavy eggs. Egg breaking strength was the highest with a value of 2.5 kg/cm2 in light eggs, whereas the thinnest eggshell (0.3862 mm) was observed in heavy eggs. pH of albumen and yolk was similar and ranged from 8.8 to 8.9 and 5.9 to 6.0, respectively. On d 14 of incubation, yolk sac weight was found higher in the medium and heavy eggs. Additionally, the dry matter of the embryo and yolk sac differed among the egg weight groups during the incubation period. Interestingly, on d 25 of incubation, the embryo weight was higher in the light and heavy eggs (35.2 and 36.3 g, respectively) than in the medium eggs (29.8 g). These findings showed that embryo growth was affected by yolk absorption and dry matter accumulation. The hatchability of total and fertile eggs was lower for the heavy eggs than the light and medium eggs. The chick weight was 42.8, 48.4, and 54.9 g in light, medium, and heavy eggs, respectively. A percentage of 34.2, 36, and 31.6% of chicks from light, medium, and heavy eggs, hatched between 637 and 648 h, 39.6, 36.2, and 32.9% between 649 and 660 h, 26.2, 27.8, and 35.5% between 661 and 672 h of incubation, respectively. In conclusion, hatching egg quality, embryo development and yolk absorption during incubation, hatch window, and hatchability were affected by egg weight in Pekin ducks.

The effect of temperature on the initial development of Brycon amazonicus Spix & Agassiz, 1829 as tool for micromanipulation of embryos.

Primordial germ cell (PGC) transplant is a promising tool in aquaculture; however, successful use of this technique requires in depth knowledge of the early stages of embryo and larval development. The aim of this study was to analyse the effect of different temperatures (22, 26, and 30°C) on the early development of B. amazonicus. The newly fertilized eggs were distributed into tanks with controlled temperature and oxygenation. Samples were collected at pre-established times and analysed under light and fluorescence microscopy. Temperature influenced the speed and duration of each stage of early development, including hatching time. The highest pronuclei fusion rate was observed 8 min post-fertilization (mpf) at 22 and 26°C, and 6 mpf at 30°C. The duration of the 512-1000 blastomeres phase during in the blastocyst stage was 1 h 30 min at 22°C, and 25 min at 26 and 30°C. Hatching occurred at 24 h 30 mpf at 22°C, 16 h post-fertilization (hpf) at 26°C, and 11 h 30 mpf at 30°C. The rate of morphologically normal larvae was 88.34% at 22°C, 90.49% at 26°C, and 73% at 30°C. Malformations of the head, yolk sac, heart, and tail were observed in all temperatures. Nevertheless, B. amazonicus embryos were able to develop satisfactory in all three temperatures tested. These results enable embryo manipulation at different temperatures to optimize the micromanipulation time of embryos and larvae for biotechnological studies.