Increased Embryo Mortality Research Articles

TPT disrupts early embryonic development and glucose metabolism of marine medaka in different salinites

Triphenyltin (TPT) is an organotin compound frequently detected in coastal estuaries, yet studies on TPT's effects in regions with significant salinity fluctuations, such as coastal estuaries, are currently limited. To investigate the toxic effects of TPT under different salinity conditions, this study focused on marine medaka (Oryzias melastigma) embryos. Through early morphological observations, RNA-seq analysis, biochemical marker assays, and qPCR detection, we explored the impact of TPT exposure on the early embryonic development of marine medaka under varying salinities. The study found that TPT exposure significantly increased embryo mortality at salinities of 0 ppt and 30 ppt. RNA-seq analysis revealed that TPT primarily affects glucose metabolism and glycogen synthesis processes in embryos. Under high salinity conditions, TPT may inhibit glucose metabolism by suppressing glycolysis and promoting gluconeogenesis. Furthermore, TPT exposure under different salinities led to the downregulation of genes associated with the insulin signaling pathway (ins, insra, irs2b, pik3ca, pdk1b, akt1, foxo1a), which may be linked to suppressed glucose metabolism and increased embryonic mortality. In summary, TPT exposure under different salinities affects the early development of marine medaka embryos and inhibits glucose metabolism. This study provides additional data to support research on organotin compounds in coastal estuaries.

Number of Blastodisc Cells During the Laying Period for Two Successive Generations of Layer and Broiler Breeder.

In the poultry industry, genetic selection for growth performance is associated with poor reproductive efficiency and an increase in embryo mortality. The identification of new biomarkers is essential to improve these parameters. The blastodisc, composed of blastodermal cells, undergoes cellular events to achieve embryo development. Factors such as hen's age, temperature and time of egg storage could influence the number of blastodermal cells and impair embryo development. In this study, we investigated the variability of the number of viable cells of blastodisc (NVCB) that could be dependent on the stage of laying and on the breed and potentially associated with reproductive parameters. In experimental breeds, eggs were collected during the whole cycle of laying. Then, the protocol was repeated on industrial breeds (breeder hens) during five successive days at three stages of laying (before, after laying peak and at the end of laying period) for two generations (mothers and offsprings). For each egg, the blastodisc was dissected in order to count viable cells. For both experiments, the NVCB increased during the laying cycle. The NVCB was higher in broiler blastodisc compared to layer blastodisc for both generations. For layer breed, the NVCB were negatively correlated with laying rate for the first generation while positively associated for offsprings. However, the NVCB was positively correlated with laying rates in both generations for broiler hens and with fertility and hatchability rates. The NVCB from fresh oviposited fertilised eggs could be a potential tool in predicting on reproductive performances in poultry.

High thermal tolerance of egg clutches and potential adaptive capacity in green turtles

Climate warming threatens sea turtles, among other effects, because high temperatures increase embryo mortality. However, not all species and populations are expected to respond the same way because they could have different thermal tolerances and capacities to adapt. We tested the effect of incubation temperature on egg mortality in a population of green turtles (Chelonia mydas) previously suggested to be less affected by extreme climatic events than others. We (1) assessed the relationship between temperature and hatching success, (2) defined an optimal range of temperatures that maximized hatching success and (3) assessed the variability in the response to temperature among clutches laid by different mothers, which could allow adaptation. Hatching success was consistently high in green turtle clutches with a skew toward high values, with 50 % of clutches having a success above 94 %. Yet, it was mildly affected by temperature, declining at both low and high temperatures. The optimal range of mean incubation temperatures was between ~30.5 °C and 32.5 °C. Current mean temperatures (31.3 °C) fall within the middle of the optimal range, indicating a potential resilience to further rises in mean nest temperature. Hatching success was best described by nest temperature and the interaction between female identity and temperature. This last predictor indicated a variability in thermal tolerance among clutches laid by different mothers and therefore, a capacity to adapt. The studied population of green turtles seems to be less vulnerable than others to climate warming. Understanding how different populations could respond to increasing temperatures could help complete the picture on the potential effects of climate change on sea turtles.

The impact of electromagnetic fields generated by high-voltage power lines on the spatial arrangement of pike (Esox Lucius Linnaeus 1758) embryos

Urbanization and technological advancements result in the dispersion of antropogenic electromagnetic fields (EMF) that can affect on ecosystems. Therefore, it is important to understand their impact on the environment. Aquatic ecosystems are subject to EMF as part of various electricity sources, e.g., high-voltage transmission lines (HVTL). We examined the impact of EMF generated by HVTL on the spatial arrangement and survival of pike (Esox lucius) embryos. Fertilized eggs were incubated under two HVTL configurations 110 kV and 220 kV compared with a control group devoid of anthropogenic EMF. Embryo orientation and survival were monitored until blastopore closure. The control group showed dominance in the arrangement of embryos along the N-S, NNW-SSE, and NNE-SSW axes, with a slight prevalence of northern directions. EMF originating from HVTL did not exert a significant influence on the spatial arrangement of pike embryos, although some deviations from the arrangement noticed in the control group were observed. Increased embryo mortality was observed only at 110 kV site, but probably due to factors unrelated to EMF. In conclusion, EMF generated by HVTL did not significantly change pike embryo orientation or chances of survival. However, longer exposure or higher EMF levels could provoke notable reactions, requiring ongoing evaluation as power networks continue to spread more widely.

Hypobaric hypoxia causes low fecundity in zebrafish parents and impairment of skeletal development in zebrafish embryos and rat offspring

Hypobaric Hypoxia (HH) negatively affects the cardiovascular and respiratory systems as well as gonadal development and the therefore next generation. This study investigated the effects of HH on zebrafish and SD rats, by exposing them to a low-pressure environment at 6000 m elevation for 30 days to simulate high-altitude conditions. It was indicated that parental zebrafish reared amh under HH had increased embryo mortality, reduced hatchability, and abnormal cartilage development in the offspring. Furthermore, the HH-exposed SD rats had fewer reproductive cells and smaller litters. Moreover, the transcriptome analysis revealed the down-regulation of steroid hormone biosynthesis pathways. The expression of the gonad-associated genes (amh, pde8a, man2a2 and lhcgr), as well as the gonad and cartilage-related gene bmpr1a, were also down-regulated. In addition, Western blot analysis validated reduced bmpr1a protein expression in the ovaries of HH-treated rats. In summary, these data indicate the negative impact of HH on reproductive organs and offspring development, emphasizing the need for further research and precautions to protect future generations' health.

Expression of chemerin and its receptors in extra-embryonic annexes and role of chemerin and its GPR1 receptor in embryo development in layer and broiler hens

Intensive genetic selection of broiler breeders and layer hens resulted in differences in the mechanisms of growth and also in cell metabolism during embryogenesis. Previous research has shown that an adipokine named chemerin and one of these receptors, CMKLR1 were potentially involved in broiler embryo development. Here, our objectives were 1. to compare the expression of chemerin and its receptors CMKLR1, GPR1 and CCRL2 and chemerin concentration in extra-embryonic annexes (allantoic and amniotic membranes and fluids and plasma) in broiler and layer fertile eggs during the development (embryonic day (ED) 7, 14 and 18) by RT-qPCR and specific chicken ELISA and 2. to investigate the role of chemerin and one of its receptors GPR1 in embryo development after in ovo injections of neutralizing antibodies against chicken chemerin and GPR1. We found that chemerin expression in amniotic membranes was higher in layer than broiler eggs at ED7 and ED14 whereas the expression of the three receptors was higher in layer than broiler in the allantoic membranes at ED14 and ED18. Chemerin concentration was more important in layer than broiler at ED14 and ED18 in amniotic liquid and at all the studied stages in blood plasma. We also showed positive correlation between amniotic chemerin concentration and chemerin amniotic membrane expression, chemerin plasma concentration and embryo body weight in both breeds. Finally, in ovo injection of chicken chemerin and GPR1 neutralizing antibodies increased embryo mortality in both layer and broiler eggs. Taken together, even if chemerin concentration and chemerin system expression in embryonic membranes are mainly higher expressed in layer than in broiler, chemerin potentially through GPR1 could promote embryo development in both breeds.

Research Note: Effects of feeding corn naturally contaminated with aflatoxin B1, deoxynivalenol, and zearalenone on reproductive performance of broiler breeders and growth performance of their progeny chicks

To evaluate the toxic effects of mycotoxin-contaminated corn (MC) on the breeders and their progeny chicks, a total of 480 fifty-wk-old Cobb broiler breeder hens were fed the following dies: 1) a corn-soybean meal diet (Control; containing 70.35% corn), 2) MC substituting for 50% of corn in Control (LM), 3) LM diet plus 2 g/kg 1 mycotoxin sequestrant, Toxy-Nil Plus (TNP) (LMT2.0), 4) MC substituting for 100% of corn in Control (HM), 5) HM diet plus 2 g/kg TN (HMT2.0), and 6) HM diet plus 2.5 g/kg TNP (HMT2.5). The MC contained 69.25 μg aflatoxin B1 (AFB1)/kg, 4,875 μg deoxynivalenol (DON)/kg, and 2,262 μg zearalenone (ZEN)/kg. At wk 4 after MC inclusion, all eggs laid were used for hatch, and all progeny chicks were fed the same mycotoxin-untreated diet for 14 d. Dietary MC inclusion decreased the hatchability of set eggs and increased embryo mortality during d 18 to 21.5. The TNP addition increased these aforementioned indices in MC-included diets. Maternal HM treatment decreased the BW of progeny chicks at age of 14 d and BWG of progeny chicks during d 1 to 14, whereas maternal LM treatment did not affect these indices. In parallel, maternal HM treatment decreased the concentrations of serum IgA, IgG, and lysozyme in the progeny chicks on d 14, but maternal LM treatment did not affect these indices. Overall, maternal dietary TNP treatments increased the growth of progeny chicks and had a trend to increase the concentrations of serum IgA and IgG on d 14 compared to maternal MC treatments. It was concluded that the feeding of relative high ratio of corn contaminated with low level of AFB1, DON, and ZEN negatively affected the reproductive performance of breeders and the growth performance of their progeny chicks, and TNP addition alleviated these toxic effects.

Embryotoxicity and Teratogenicity of Steroidal Saponin Isolated from Ophiopholis mirabilis.

Benthic invertebrates produce secondary metabolites that serve as defenses against consumers and promote their fitness. To explore the chemical defense in marine benthic echinoderms, the chemical constituents of Ophiopholis mirabilis were investigated. A steroidal monoglycoside, asterosaponin P1, was isolated from O. mirabilis for the first-time using column chromatography. The chemical structure was characterized by spectroscopic and spectrometric methods. The embryotoxicity and teratogenicity of the isolated compound were assessed using the zebrafish embryo assay, a powerful vertebrate animal model system to study mechanisms of toxicity. When applied at high concentrations, asterosaponin P1 causes a significant increase in embryo mortality. A moderate LC50 of asterosaponin P1 appeared to be time- and concentration-dependent in its toxicity to zebrafish embryos. Teratogenicity in zebrafish embryos also included morphological defects, decreased hatchability, and a reduced heart rate. These findings revealed that steroidal saponin extracted from O. mirabilis exhibited acute toxic effects on zebrafish embryos, suggesting a potential chemical defense function in marine habitats.

The Development of the Bacterial Community of Brown Trout (Salmo trutta) during Ontogeny.

Brown trout (Salmo trutta) is an important aquaculture species in Germany, but its production faces challenges due to global warming and a high embryo mortality. Climate factors might influence the fish's bacterial community (BC) and thus increase embryo mortality. Yet, knowledge of the physiological BC during ontogeny in general is scarce. In this project, the BC of brown trout has been investigated in a period from unfertilized egg to 95 days post fertilization (dpf) using 16S rRNA gene amplicon sequencing. Developmental changes differed between early and late ontogeny and major differences in BC occurred especially during early developmental stages. Thus, analysis was conducted separately for 0 to 67 dpf and from 67 to 95 dpf. All analyzed stages were sampled in toto to avoid bias due to different sampling methods in different developmental stages. The most abundant phylum in the BC of all developmental stages was Pseudomonadota, while only two families (Comamonadaceae and Moraxellaceae) occurred in all developmental stages. The early developmental stages until 67 dpf displayed greater shifts in their BC regarding bacterial richness, microbial diversity, and taxonomic composition. Thereafter, in the fry stages, the BC seemed to stabilize and changes were moderate. In future studies, a reduction in the sampling time frames during early development, an increase in sampling numbers, and an attempt for biological reproduction in order to characterize the causes of these variations is recommended.

Influence of hypomagnetic field on the heartbeat in zebrafish embryos.

The magnetic environment may influence the functioning of the cardiovascular system. It was reported that low-frequency and static magnetic fields affect hemodynamics, heart rate, and heart rate variability in animals and humans. Moreover, recent data suggest that magnetic fields affect the circadian rhythms of physiological processes. The influence of the magnetic environment on heart functionating during early development has been studied insufficiently. We utilized transparent zebrafish embryos to evaluate the effect of the hypomagnetic field on the characteristics of cardiac function using a noninvasive optical approach based on photoplethysmographic microscopic imaging. The embryos were exposed to the geomagnetic and hypomagnetic fields from the second to the 116th hour post fertilization under a 16h light/8h dark cycle or constant illumination. The exposure of embryos to the hypomagnetic field in both lighting modes led to increased embryo mortality, the appearance of abnormal phenotypes, and a significant increase in the embryo's heartbeat rate. The difference between maximal and minimal heartbeat intervals, maximal to minimal heartbeat intervals ratio, and the coefficient of variation of heartbeat rate were increased in the embryos exposed to the hypomagnetic field under constant illumination from 96 to 116h post fertilization. The dynamics of heartbeat rate changes followed a circadian pattern in all studied groups except zebrafish exposed to the hypomagnetic field under constant illumination. The results demonstrate the importance of natural magnetic background for the early development of zebrafish. The possible mechanisms of observed effects are discussed.

Lamprey immune protein triggers the ferroptosis pathway during zebrafish embryonic development

BackgroundPreviously, a novel lamprey immune protein (LIP) was identified, which plays an important role in immunity and the regulation of growth and development in lampreys. However, the mechanism of how LIP regulates growth and development remains unclear.MethodsIn this study, a zebrafish model of LIP overexpression was established by delivering a transgenic plasmid to the fertilized egg. The biological function of LIP was explored in vivo through phenotypic characterization, comparative transcriptome sequencing, and physiological and biochemical analyses.ResultsLIP caused developmental toxicity in zebrafish, increased embryo mortality and exhibited strong teratogenic, lethal, and developmental inhibitory effects. Comparative transcriptome analysis showed that LIP-induced large-scale cell death by triggering ferroptosis. Furthermore, LIP-induced lipid peroxidation and caused pericardial edema. Direct inhibition of acsl4a and tfr1a, or silencing of acsl4a and tfr1a with specific siRNA suppressed ferroptosis and pericardial edema.ConclusionsTaken together, we confirmed that LIP can participate in growth and development via the regulation of lipid peroxidation and ferroptosis. This lays the foundation for future studies on the function of LIP in lampreys.FULq97gHbZiSwv22VJ3jSJVideo

In ovo exposure to cadmium causes right ventricle hyperplasia due to cell proliferation of cardiomyocytes

Cadmium (Cd) is an environmental and occupational pollutant inhaled through smoking or ingested through contaminated food. Yet, little is known about its teratogenicity. In this study, the effects of Cd on embryonic heart development were investigated by exposing Cd to chicken embryos in ovo. Fertilized eggs were treated with Cd at Hamburger-Hamilton Stage (HH)16 and collected at HH35 for histological evaluation of the heart. Cd treatment of 100 μM at HH16 increased embryo mortality at HH35. Specific structural heart defects were not observed in any Cd treatment group, but the relative myocardial tissue area of the right ventricle was increased with Cd exposure.When the HH31 hearts were stained with p-H3S10, the right ventricle had an increased number of cells undergoing proliferation, which was associated with upregulation of Cdk1, Cdk6, CycA, CycD, and CycE detected by qPCR. These findings suggest that Cd exposure from HH16 upregulates proliferation genes and drives overgrowth of the right ventricle. These results grant further attention to Cd teratogenicity on embryonic heart development. Such morphological changes in the heart can potentially affect cardiac function and increase the risk for future cardiovascular diseases, such as heart failure.

Chemerin is secreted by the chicken oviduct, accumulates in egg albumen and could promote embryo development

Understanding of the distribution of chemerin and its receptors, Chemokine-like Receptor 1 (CMKLR1), G Protein-coupled Receptor 1 (GPR1) and Chemokine (C–C motif) receptor-like 2 (CCRL2), in the egg and the embryonic annexes is currently lacking, and their role during embryogenesis remains unknown. By immunoblot using monoclonal anti-chicken antibodies and Enzyme Linked Immunosorbent Assays (ELISA), we found that chemerin is expressed 10 times higher in albumen eggs than in blood plasma, and it is also abundant in the perivitelline membrane but undetectable in yolk. Chicken chemerin can inhibit bacterial growth. By Reverse Transcription—quantitative Polymerisation Chain Reaction (RT-qPCR), western-blot, and immunofluorescence, we show that chemerin is locally produced by the oviduct magnum that participates in albumen formation. Using cultures of magnum explants, we demonstrate that progesterone (P4) and oestradiol (E2) treatment increases chemerin secretion into cultured media and expression in magnum. Chemerin and its three receptors are present in amniotic and Chorio Allantoic Membranes (CAM). Only CMKLR1 expression decreased from embryonic day (ED) 7 to ED11 and remained low until ED18. Chemerin concentrations strongly increased in amniotic fluid at D14 when egg albumen crossed the amniotic membrane. In ovo injections of neutralising chemerin and CMKLR1 antibodies (0.01, 0.1 and 1 µg) increased embryo mortality, which occurred mainly at ED12-13, in a dose-dependent manner. Chemerin treatment increased primary CAM viability. Finally, chemerin and CMKLR1 inhibition within the CAM led to a decrease in blood vessel development and associated angiogenic gene expression. Our results show an important function of the chemerin system during embryo development in chickens, suggesting the potential use of this adipokine as a predictive marker for egg fertility or hatchability.

Frontiers in sheep reproduction - making use of natural responses to environmental challenges to manage productivity.

This review addresses advances, directions and opportunities for research on sheep reproduction in the context of the global challenges of food security and climate change, and demand for 'clean, green and ethical' (CGE) animal management. The foundation of CGE management is an understanding of the physiological processes through which the reproductive system responds to changes in the animal's environment. These days, to the main environmental factors (photoperiod, nutrition, pheromones), we need to add stress from extreme weather events. With respect to nutrition in rams, we now have a deeper understanding of the responses of the brain centres that control gonadotrophin secretion (the kisspeptin system). At testis level, we have found that nutrition affects non-coding RNAs in Sertoli cells and germ cells, thus affecting the balance between cell proliferation and apoptosis. This proliferation-apoptosis balance is also affected during prenatal development, when undernutrition or stress in pregnant ewes seems to elicit epigenetic changes in developing gonads that could affect offspring fertility in adult life. With respect to nutrition in ewes, metabolic signals act directly on ovarian follicles, and thus change ovulation rate, but the variety of signals now includes the adipokines. An early concern was that nutritional supplements that increase ovulation rate would also increase embryo mortality but we now know that embryo survival is improved under field conditions. Finally, we had always thought that the efficiency gains from early puberty in lambs could only be achieved by accelerating fat accumulation, but we now know that faster muscle growth will achieve the same goal, offering two advantages in meat production systems. With respect to pheromones ('ram effect'), we have a deeper understanding of the brain responses (kisspeptin system) but, most importantly, a realization that the response of ewes to the ram signal involves cell division in memory centres. Many opportunities remain.

Teratogenic of 2,3,7,8-Tetrachlorodibenzo-P-Dioxin (TCDD) on Eye Diameters and Body Weight in Chicken Embryo

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a chemical compound resulting from the burning effect that is very dangerous for humans, plants, or animals which can cause skin damage, immunotoxic, hepatotoxic, carcinogenic, teratogenic, effects on reproduction, growth processes, neurobehavioral, and endocrine effects that are specific In addition, this compound can also reduce embryo weight and increase embryo mortality. This study aimed to determine the teratogenic of TCDD administration on eye diameters and body weight of chicken embryo. In this study there were 2 treatment groups, i.e: 1) P1 as a normal group without the addition of TCDD, and 2) P2 as a group with the addition of TCDD of 5 ng / egg with dissolved in 5 µL corn oil. The treatment was carried out for 7 days. The measurement data of eye diameters and body weight of chicken embryo were analyzed by the unpaired sample T test. The analysis showed that the teratogenic of TCDD had a significant effect on eye diameters and body weight of chicken embryos, where the teratogenic of TCDD caused a decrease in eye diameters and body weight loss in chicken embryos.

Chorion Alterations in Eyed-Stage Salmonid Eggs Farmed in La Araucanía, Chile: A Retrospective Study.

Simple SummaryThe chorion (also called egg envelope) is the primary envelope that protects the fish embryo against drying, mechanical actions, and abrupt changes in the water conditions. Alterations of the chorion during the embryo incubation are not unusual, but these are scarcely reported. Increased occurrence of chorion alterations can lead to decreased reproductive performance and important losses for fish farms. Here, we described several chorion alterations observed in samples of embryonated eggs collected from different salmon and trout farms located in southern Chile over a period of 14 years. We detected four types of chorion alterations and found soft chorion as the most prevalent alteration in the years analyzed, affecting mainly Atlantic and Coho Salmon. Eggs of Rainbow Trout displayed fewer chorion alterations among the three species analyzed. As the eggs analyzed here were produced under standard industrial conditions, we conclude that these alterations are possibly linked to changes in water conditions, which need to be further investigated.The chorion is the primary envelop that protects the fish embryo against mechanical actions, pathogens, and abrupt changes in physical and chemicals conditions of the incubation medium. During embryo development, chorion alterations are not rare, but the occurrence of these is scarcely reported. Increased frequency of chorion alterations can result in increased embryo mortality and thus decreased reproductive performance and losses for fish farms. In this study, we characterize different chorion alterations observed in samples collected over 14 years from 12 salmon and trout farms located in the region of La Araucanía in southern Chile, which sent live eyed-stage embryos (‘eyed-eggs’) for quality analysis to our laboratory. We found soft chorion as the most common alteration observed, being present in the whole 14-year series analyzed in Atlantic Salmon (Salmo salar) and affecting up to 35.0% of the samples examined in a year. This alteration also affected up to 20.0 and 5.7% of Coho Salmon (Oncorhynchus kisutch) and Rainbow Trout (Oncorhynchus mykiss) samples analyzed in a year, respectively. We also found an increase of other chorion alterations, including perforated and white-spotted chorion in Atlantic and Coho Salmon, in the last 8 years. Among the three species, Rainbow Trout exhibited fewer chorion alterations. As the embryonated eggs analyzed here were obtained from broodstocks maintained under standard industrial conditions, these alterations might be linked to changes in environmental conditions affecting the incubation water that need to be further investigated.

Effect of egg storage duration and egg turning during storage on egg quality and hatching of broiler hatching eggs

In commercial hatcheries, it is common to store eggs before incubation. One practice to improve hatchability consists in egg turning during this storage. This work aims to highlight the effects of turning on the physicochemical aspects of eggs and, consequently, how this turning can influence the hatching of chicks. An experiment was conducted to evaluate the effects of storage duration and egg turning during storage on egg quality, hatchability, and residual analysis. A total of 7 500 hatching eggs were collected from a 55-week-old commercial Cobb500 breeder flock and storage according to the treatments. The experiment was completely randomized in a 3×2 factorial design with three storage periods (4, 8, and 12 days) and egg turning (180° turn of eggs once a day) or no turning during storage, totaling six treatments. Regardless of turning, eggs stored for 4 days weighed more than turned eggs stored for 8 and 12 days, which were similar (P < 0.05). Non-turned eggs experienced an increase in relative shell weight with increased storage duration, and non-turned eggs stored for 4 and 8 days differed from non-turned eggs stored for 12 days (P < 0.05). Albumen pH of turned eggs stored for 4 and 8 days was lower than that of non-turned eggs stored for the same durations (P < 0.05). Albumen pH of turned eggs increased as storage duration increased (P < 0.05). Egg turning increased hatching by 2.02% over that of non-turning (P < 0.05). Eggs stored for 12 days, irrespective of turning, had higher late embryonic mortality (P < 0.05) compared to the other treatments. It was concluded that turning eggs during pre-incubation storage was adequate to improve hatchability of fertile eggs. Storing fertile eggs for 12 days is harmful to egg quality and increases embryo mortality even if eggs were turned.

Ocean Acidification and Ocean Warming Effects on Pacific Herring (Clupea pallasi) Early Life Stages

Increasing green house gas emissions are expected to raise surface seawater temperatures and lead to locally intensified ocean acidity in the U.S. Pacific Northwest. Pacific herring (Clupea pallasi) are ecologically and economically important forage fish species native to this region. While the impacts of ocean acidification and ocean warming on organism physiology have been extensively studied, less is known on how concurrent climate change stressors will affect marine fish. Therefore, our study focused on the combined effects of ocean acidification and warming on Pacific herring early life history stages. Pacific herring embryos were incubated under a factorial design of two temperature (10°C or 16°C) and twopCO2(600 μatm or 1200 μatm) treatments from fertilization until hatch (6 to 15 days depending on temperature). ElevatedpCO2was associated with a small increase in embryo mortality. Elevated temperature, as a single stressor, generated greater embryo mortality and embryo heart rates, larger yolk areas upon hatch, lower hatching success, and shorter larval lengths; compared with the same parameters measured under ambient temperature. The interaction of elevated temperature andpCO2was associated with greater embryo heart rates and yolk areas compared to ambient conditions. This study suggests that while temperature is the primary global change stressor affecting Pacific herring embryology, interaction effects withpCO2could introduce additional physiological challenges.

Maternal hepcidin determines embryo iron homeostasis in mice

AbstractIron disorders are associated with adverse pregnancy outcomes, yet iron homeostatic mechanisms during pregnancy are poorly understood. In humans and rodents, the iron-regulatory hormone hepcidin is profoundly decreased in pregnant mothers, which is thought to ensure adequate iron availability for transfer across placenta. However, the fetal liver also produces hepcidin, which may regulate fetal iron endowment by controlling placental iron export. To determine the relative contribution of maternal vs embryo hepcidin to the control of embryo iron endowment in iron-sufficient or iron-overloaded mice, we generated combinations of mothers and embryos that had or lacked hepcidin. We found that maternal, but not embryonic, hepcidin determined embryo and placental iron endowment in a healthy pregnancy. We further determined that inflammation can counteract pregnancy-dependent suppression of maternal hepcidin. To establish how essential maternal hepcidin suppression is for embryo iron homeostasis, we mimicked the range of maternal hepcidin activity by administering a hepcidin peptide mimetic to pregnant mice. This also allowed us to determine the effect of isolated maternal hepcidin excess on pregnancy, in the absence of other confounding effects of inflammation. Higher doses of hepcidin agonist caused maternal iron restriction and anemia, lower placenta and embryo weight, embryo anemia, and increased embryo mortality. Low agonist doses did not cause maternal anemia but still adversely affected the embryo, causing anemia, tissue iron deficiency (including in the brain), and decreased weight. Our studies demonstrate that suppression of maternal hepcidin during pregnancy is essential for maternal and embryo iron homeostasis and health.

Developmental toxicity of Clerodendrum cyrtophyllum turcz ethanol extract in zebrafish embryo

Ethnopharmacological relevanceClerodendrum cyrtophyllum Turcz has been used in traditional medicine for the treatment of various diseases. In spite of its therapeutic applications, research on its toxicity and teratogenicity is still limited. Aim of the studyThe study aimed to investigate the developmental toxicity of the ethanol extract of C. cyrtophyllum (EE) in zebrafish embryo model. Material and methodsMajor compounds from crude ethanol extract of Clerodendron cyrtophyllum Turcz leaves were determined using HPLC-DAD-Orbitrap-MS analysis. The developmental toxicity of EE were investigated using zebrafish embryo model. Zebrafish embryos at 6 h post-fertilization (hpf) were treated with EE at different concentrations. Egg coagulation, mortality, hatching, yolk sac edema, pericardial edema and teratogenicity were recorded each day for during a 5-day exposure. At time point 120 hpf, body length, pericardial area, heartbeat and yolk sac area were assessed. In order to elucidate molecular mechanisms for the developmental toxicity of EE, we further evaluated the effects of the EE on the expression of genes involved on signaling pathways affecting fish embryo’s development such as heart development (gata5, myl7, myh6, has2, hand2, nkx 2.5), oxidative stress (cat, sod1, gpx4, gstp2), wnt pathway (β-catenin, wnt3a, wnt5, wnt8a, wnt11), or cell apoptosis (p53, bax, bcl2, casp3, casp8, casp9, apaf-1, gadd45bb) using qRT-PCR analysis. ResultsOur results demonstrated that three major components including acteoside, cirsilineol and cirsilineol-4'-O-β-D-glucopyranoside were identified from EE. EE exposure during 6–96 h post-fertilization (hpf) at doses ranging from 80 to 200 μg/mL increased embryo mortality and reduced hatching rate. EE exposure at 20 and 40 μg/mL until 72–120 hpf induced a series of malformations, including yolk sac edema, pericardial edema, spine deformation, shorter body length. Based on two prediction models using a teratogenic index (TI), a 25% lethality concentration (LD25) and the no observed-adverse-effect level (NOAEL), EE is considered as teratogenic for zebrafish embryos with TI (LC50/EC50) and LD25/NOAEC values at 96 hpf reaching 3.87 and 15.73 respectively. The mRNA expression levels of p53, casp8, bax/bcl2, gstp2, nkx2.5, wnt3a, wnt11, gadd45bb and gata5 were significantly upregulated by EE exposure at 20 and 40 μg/mL while the expression of wnt5, hand2 and bcl2 were downregulated. ConclusionsThese results provide evidence for toxicity effects of EE to embryo stages and provide an insight into the potential toxicity mechanisms on embryonic development.