Embryo Weight Research Articles

Oenothera biennis improves pregnancy outcomes by suppressing inflammation and fibrosis in an intra-uterine adhesion rat model

Intrauterine adhesion (IUA), also referred to as Asherman’s syndrome, is characterized by fibrosis, inflammation, and can cause amenorrhea and infertility due to abnormal endometrial healing. Histological and Molecular methods were used to evaluate the efficacy of EPO, which is traditionally known for its anti-inflammatory and fibrinolytic properties, in preventing the formation of IUA. Oral administration of EPO reduced the formation of adhesion bands and promoted endometrial regeneration. EPO administration decreased extracellular matrix accumulation, evidenced by the down-regulation of tissue COL1A1 and COL3A1 expression. The anti-inflammatory effect of EPO was confirmed by a reduction in oxidants and down-regulation of pro-inflammatory cytokines including TNF-α, IL-6, IFN-γ, and IL-1β. Furthermore, EPO improved embryonic development parameters, including size and weight of embryo, as well as increased embryo count and live embryo percentage in the rat IUA model. EPO also positively enhanced implantation markers, particularly enlargement and mass gain in the placenta of the treated group, consequently improving pregnancy outcomes such as the number of babies, percent of live babies, baby weight and gestation time. Histopathological investigation provides evidence that oral administration of EPO showed no toxicity on the main three organs including liver, kidney and heart. These results showed that EPO can be considered as a safe and natural product with potent anti-inflammatory and fibrinolytic properties without any observed side effects for the treatment of IUA.

In-utero transfer of decidualized endometrial stromal cells increases the frequency of regulatory T cells and normalizes the abortion rate in the CBA/J × DBA/2 abortion model.

Failure to adequate decidualization leads to adverse pregnancy outcomes including pregnancy loss. Although there are plenty of reports underscoring immune dysfunction as the main cause of abortion in CBA/J females mated with DBA/2 males (CBA/J × DBA/2), little is known about the potential role of impaired endometrial decidualization. Endometrial stromal cells (ESCs) from CBA/J mice were in-vitro decidualized, and the proteome profile of the secretome was investigated by membrane-based array. CBA/J mice were perfused In-utero with either decidualized ESCs (C×D/D), undecidualized ESCs (C×D/ND), or PBS (C×D/P) 12 days before mating with DBA/2 males. Control mice were not manipulated and were mated with male DBA/2 (C×D) or Balb/c (C×B) mice. On day 13.5 of pregnancy, reproductive parameters were measured. In-vivo tracking of EdU-labeled ESCs was performed using fluorescence microscopy. The frequency of regulatory T cells (Tregs) in paraaortic/renal and inguinal lymph nodes was measured by flow cytometry. The proliferation of pregnant CBA/J splenocytes in response to stimulation with DBA/2 splenocytes was assessed by 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) flow cytometry. In C×D/D mice, the resorption rate was reduced to match that seen in the C×B group. Intrauterine perfused ESCs appeared in uterine stroma after 2 days, which remained there for at least 12 days. There was no difference in the number of implantation sites and embryo weight across all groups. The frequency of Tregs in the inguinal lymph nodes was similar across all groups, but it increased in the paraaortic/renal lymph nodes of C×D/D mice to the level found in C×B mice. No significant changes were observed in the proliferation of splenocytes from pregnant C×D/D compared to those of the C×D group in response to stimulation with DBA/2 splenocytes. Decidualization of ESCs was associated with a profound alteration in ESC secretome exemplified by alteration in proteins involved in extracellular matrix (ECM) remodeling, response to inflammation, senescence, and immune cell trafficking. Our results showed that the deficiency of Tregs is not the primary driver of abortion in the CBA/J × DBA/2 model and provided evidence that impaired endometrial decidualization probably triggers endometrial immune dysfunction and abortion in this model.

Sex-specific DNA methylation and gene expression changes in mouse placentas after early preimplantation alcohol exposure

During pregnancy, exposure to alcohol represents an environmental insult capable of negatively impacting embryonic development. This influence can stem from disruption of molecular profiles, ultimately leading to manifestation of fetal alcohol spectrum disorder. Despite the central role of the placenta in proper embryonic development and successful pregnancy, studies on the placenta in a prenatal alcohol exposure and fetal alcohol spectrum disorder context are markedly lacking. Here, we employed a well-established model for preimplantation alcohol exposure, specifically targeting embryonic day 2.5, corresponding to the 8-cell stage. The exposure was administered to pregnant C57BL/6 female mice through subcutaneous injection, involving two doses of either 2.5 g/kg 50 % ethanol or an equivalent volume of saline at 2-hour intervals. Morphology, DNA methylation and gene expression patterns were assessed in male and female late-gestation (E18.5) placentas. While overall placental morphology was not altered, we found a significant decrease in male ethanol-exposed embryo weights. When looking at molecular profiles, we uncovered numerous differentially methylated regions (DMRs; 991 in males; 1309 in females) and differentially expressed genes (DEGs; 1046 in males; 340 in females) in the placentas. Remarkably, only 21 DMRs and 54 DEGs were common to both sexes, which were enriched for genes involved in growth factor response pathways. Preimplantation alcohol exposure had a greater impact on imprinted genes expression in male placentas (imprinted DEGs: 18 in males; 1 in females). Finally, by using machine learning model (L1 regularization), we were able to precisely discriminate control and ethanol-exposed placentas based on their specific DNA methylation patterns. This is the first study demonstrating that preimplantation alcohol exposure alters the DNA methylation and transcriptomic profiles of late-gestation placentas in a sex-specific manner. Our findings highlight that the DNA methylation profiles of the placenta could serve as a potent predictive molecular signature for early preimplantation alcohol exposure.

Research Note: Effects of different light-emitting diode lights during egg incubation on hatching performance and embryo development in White King pigeons

The light provide during incubation can influence hatching characteristics (hatching time, hatchability, etc.) and embryo development in chickens, geese, and turkeys. However, relevant studies on this factor in pigeons are lacking. This study investigated the effects of in ovo photostimulation during embryogenesis on hatching performance, squab quality, and embryo development in pigeons. 400 eggs from paired- bred pigeons were randomly distributed into 4 incubation lighting treatments, with 2 replicates per treatment. The treatments included dark as a control (NL), 12-h light, and 12-h dark photoperiods of white light (WL), red light (RL), and green light (GL) (100 lx at egg level) during the first 15 d of incubation. A total of 1,600 eggs in 4 batches from White King pigeons were used. The results showed that hatching time of the WL group was significantly shorter than that of the dark light group (P < 0.05). The hatchability of fertile eggs in the WL group was significantly higher (P < 0.05), whereas the hatchability of fertile eggs in the RL group was significantly lower (P < 0.05) than that of in the control group. Light stimulation had no effect on time to 90% hatching or average hatching time (P > 0.05). In addition, the hatch window was not extended by light stimulation (P > 0.05). The group incubated under GL showed an increase in embryo weight and relative leg muscle on embryonic d 14 and the hatching day compared to the dark incubation (P < 0.05). Green light stimulated the heart and liver development during the early and middle stages of embryogenesis. It was concluded that white light stimulation during embryogenesis accelerated the hatching process, whereas monochromatic green light had a positive effect on embryo development. Our findings provide important guidance for developing light protocols for pigeon egg incubation.

Evaluating the effects of virulence genotype, swarming motility, and multi-locus sequence types of Escherichia coli on layer chicken embryos.

To determine the effects of swarming motility (SM) and multi-locus sequence types (MLST) on the main effect of virulence genotype of Escherichia coli through an embryos lethality assay between the 12th and 18th days of incubation. We collected 58 E. coli isolates from asymptomatic commercial hens (n=42) and lesions of colibacillosis cases (n=16), then classified their virulence genotype as avirulent, moderately virulent, virulent-healthy, and virulent-colibacillosis categories by the presence of five virulence-associated genes (iroN, ompT, hlyF, iutA, and iss). These isolates were further classified as non-motile, motile, or hyper-motile by SM assay. From the 58 isolates, we selected 29 for ELA and determined their MLST. Each isolate was inoculated into 15 embryonated eggs through the allantoic cavity. We found the avirulent isolates reduced the relative embryo weight compared to virulent-colibacillosis and moderately virulent isolates (37.49vs. 41.51 and 40.34%, P=0.03). Among the moderately virulent and virulent-colibacillosis categories, embryo lethality was lower when isolates were non-motile. Yolk retention was unaffected by virulence categories, motility, or MLST. Interaction between virulence genotype and SM substantially influenced the embryo lethality assay of E. coli isolates.

The developmental pattern related to fatty acid uptake and oxidation in the yolk sac membrane and jejunum during embryogenesis in Muscovy duck

This study aimed to investigate the developmental change of body growth and gene expression related to fatty acid uptake and oxidation in the yolk sac membrane (YSM) and jejunum during embryogenesis in Muscovy ducks. The weights of embryos and yolk sac (YS) (5 embryos per replicate, n = 6) were recorded on embryonic days (E)16, E19, E22, E25, E28, E31, and the day of hatch (DOH). The fat and fatty acid contents in YSM, jejunal histology, and gene expression related to fatty acid metabolism in YSM and jejunum were determined in each sampling time. Among the nonlinear models, the maximum growth is estimated at 2.83 (E22.5), 2.67 (E22.1), and 2.60 (E21.3) g/d using logistic, Gompertz, and Von Bertalanffy models, respectively. The weight of YS, and ether extract-free YS as well as the amounts of fat and fatty acids in YS decreased (P < 0.05) linearly, whereas the villus height, crypt depth, villus height/crypt depth, and musculature thickness in jejunum increased (P < 0.05) linearly during embryogenesis. The mRNA expression of CD36, SLC27A4, and FABP1 related to fatty acid uptake as well as the mRNA and protein expressions of PPARα and CPT1 related to fatty acid oxidation increased in a quadratic manner (P < 0.05) in both YS and jejunum, and the maximum values were achieved during E25 to E28. In conclusion, the maximum growth rate of Muscovy duck embryos was estimated at 2.60 to 2.83 g/d on E21.3 to E23.5, while the accumulations of lipid and fatty acid in YS were decreased in association with the increased absorptive area of morphological structures in jejunum. The gene and protein expression involved in fatty acid metabolism displayed a similar enhancement pattern between YSM and jejunum during E25 to E28, suggesting that fatty acid utilization could be strengthened to meet the energy demand for embryonic development.

A Comparative Analysis of the Cytotoxic and Vascular Activity Effects of Western Diamondback Rattlesnake (Crotalus atrox) and Eastern Diamondback Rattlesnake (Crotalus adamanteus) Venoms Using a Chick Embryo Model.

Crotalus snakebites induce various toxicological effects, encompassing neurological, myotoxic, and cytotoxic symptoms, with potentially fatal outcomes. Investigating venom toxicity is essential for public health, and developing new tools allows for these effects to be studied more comprehensively. The research goals include the elucidation of the physiological consequences of venom exposure and the assessment of toxicity using animal models. Chicken embryos serve as valuable models for assessing venom toxicity through the chick embryotoxicity screening test (CHEST) and the chick chorioallantoic membrane (CAM) assay, particularly useful for evaluating vascular impacts. C. adamanteus venom application resulted in higher embryotoxicity and morphological abnormalities, such as Siamese twins. The CAM assay demonstrated the hemorrhagic effects of venom, varying with venom type and concentration. The irritant potential of both venom types was classified as slight or moderate depending on their concentration. Additionally, acetylcholinesterase (AChE) activity was performed to receive information about organ toxicity. The results show that both venoms induced changes in the whole embryo, heart, and liver weights, but the C. adamanteus venom was identified as more toxic. Specific venom concentrations affected AChE activity in embryonic tissues. These findings underscore the embryotoxic and vasoactive properties of Crotalus venoms, providing valuable insights into their mechanisms of toxicity and potential applications in biomedicine.

Transcriptomic and epigenomic insights into pectoral muscle fiber formation at the late embryonic development in pure chicken lines

Long-term intensive genetic selection has led to significant differences between broiler and layer chickens, which are evident during the embryonic period. Despite this, there is a paucity of research on the genetic regulation of the initial formation of muscle fiber morphology in chick embryos. Embryonic d 17 (E17) is the key time point for myoblast fusion completion and muscle fiber morphology formation in chickens. This study aimed to explore the genetic regulatory mechanisms underlying the early muscle fiber morphology establishment in broiler chickens of Cornish (CC) and White Plymouth Rock (RR) and layer chickens of White Leghorn (WW) at E17 using the transcriptomic and chromatin accessibility sequencing of pectoral major muscles. The results showed that broiler chickens exhibited significant higher embryo weight and pectoral major muscle weight at E17 compared to layer chickens (P = 0.000). A total of 1,278, 1,248, and 892 differentially expressed genes (DEGs) of RNA-seq data were identified between CC vs. WW, RR vs. WW, and CC vs. RR, separately. All DEGs were combined for cluster analysis and they were divided into 6 clusters, including cluster 1 with higher expression in broilers and cluster 6 with higher expression in layers. DEGs in cluster 1 were enriched in terms related to macrophage activation (P = 0.002) and defense response to bacteria (P = 0.002), while DEGs in cluster 6 showed enrichment in protein-DNA complex (P = 0.003) and monooxygenase activity (P = 0.000). ATAC-seq data analysis identified a total of 38,603 peaks, with 13,051 peaks for CC, 18,780 peaks for RR, and 6,772 peaks for WW. Integrative analysis of transcriptomic and chromatin accessibility data revealed GOLM1, ISLR2, and TOPAZ1 were commonly upregulated genes in CC and RR. Furthermore, screening of all upregulated DEGs in cluster 1 from CC and RR identified GOLM1, ISLR2, and HNMT genes associated with neuroimmune functions and MYOM3 linked to muscle morphology development, showing significantly elevated expression in broiler chickens compared to layer chickens. These findings suggest active neural system connectivity during the initial formation of muscle fiber morphology in embryonic period, highlighting the early interaction between muscle fiber formation morphology and the nervous system. This study provides novel insights into late chick embryo development and lays a deeper foundation for further research.

Differential Effects of n-3 and n-6 Polyunsaturated Fatty Acids on Placental and Embryonic Growth and Development in Diabetic Pregnant Mice.

The present study aimed to investigate the differential effects of n-3 and n-6 polyunsaturated fatty acids (PUFAs) on placental and embryonic development. Pregnant mice were assigned to five groups: healthy control (HC), diabetes mellitus control (DMC), diabetes + low-dose n-3 PUFA (Ln-3), diabetes + high-dose n-3 PUFA (Hn-3), and diabetes + n-6 PUFA (n-6). On E12.5d, the Hn-3 group, but not the n-6 group, had a higher placenta weight. The weight ratio of embryo to placenta in the n-6 group was significantly lower than in the Hn-3 group but higher than in the DMC group. The Hn-3 group had significantly higher protein levels of VEGF, IGF-1, and IGFBP3, while the n-6 group had lower VEGF than the DMC group. Compared with the DMC group, embryonic Cer-16:0 was significantly higher in the Hn-3 group, while embryonic PC (36:6), PC (38:7), and PE (40:7) were significantly lower in the n-6 group. The embryo and placenta weights were positively correlated with placental VEGF, IGFBP3, and embryonic Cer-16:0, and they were negatively correlated with embryonic PC (36:6) and PE (40:7). The weight ratio of embryo to placenta was negatively correlated with embryonic PC (36:6). In addition, embryonic Cer-16:0 was positively correlated with placental VEGF and IGFBP3. In conclusion, n-3 PUFA and n-6 PUFA improved placental and embryonic growth through different mechanisms.

The significance of light chronodestruction in the development of the embryotoxic effect in the experiment

Objective. To assess the significance of light chronodestruction in the development of the embryotoxic effect in the experiment. Materials and methods. The experiment was conducted on 60 female rats of the Wistar line. To assess the significance of chronodestruction in the development of embryotoxic effect, the uterus was isolated in the experiment, in which the number of implantation sites, the total number of embryos, the number of live and dead embryos, and the number of resorptions were estimated. The number of corpus luteum in the harvested ovaries was estimated. In addition, the mean embryo weight (g) and mean craniocaudal size (mm) were recorded. Indicators of embryotoxic effects of light deprivation were determined by pre- and post-implantation death, which was calculated in accordance with methodological recommendations. Results. Exposure to light deprivation reduces the number of implantation sites as of 7th, 14th and 21st days by 1,5-1,8 times, the total number of embryos by 1,6-1,8 times, the number of living embryos by 2,2-9 times and increases the number of dead embryos as of 14th and 21st days by 4-5,5 times, the number of resorptions – 1,6-11 times. The average craniocaudal size (mm) of embryos in experimental animals was recorded below control values by 1,5 times, 1,3 and 3,7 times by 7th, 14th, 21st days respectively. In females exposed to light deprivation, there was a significant increase in pre-implantation mortality by 35-41,8 times and post-implantation mortality by 7,2-20,4 times compared to the control. Conclusion. Light deprivation may have a negative effect on pregnancy and fetal development in female rats, which is confirmed by an increase in pre- and post-implantation mortality.

Placenta-anchored tadalafil liposomes rescues intrauterine growth restriction through continuous placental blood perfusion improvement

Physiological or pathological hypoperfusion of the placenta is one of the main causes of intrauterine growth restriction (IUGR) which poses a significant risk to the health of the fetus and newborn. Tadalafil, a 5-type phosphodiesterase inhibitor, has previously been found to improve the symptoms of IUGR in various clinical studies. Unfortunately, its clinical utility is hindered by its limited water solubility, rapid metabolism, and lack of specific distribution in target tissues rendering tadalafil unable to maintain long-term placental perfusion. In this study, iRGD-modified tadalafil-loaded liposomes (iRGD-lipo@Tad) featuring a size of approximately 480 nm were designed to rectify the shortcomings of tadalafil. The prepared iRGD-lipo@Tad exhibited superior stability, sustained drug release capacity, and low cytotoxicity. The fluorescence study, tissue slice study, and drug biodistribution study together demonstrated the placenta-anchored ability of iRGD-modified liposomes. This was achieved by a dual approach consisting of the iRGD-mediated placenta-targeting effect and special particle size-mediated placenta resident effect. The pharmacokinetic study revealed a significant improvement in the in vivo process of tadalafil encapsulated by the iRGD-modified liposomes. In comparison to the tadalafil solution, the peak plasma concentration of iRGD-lipo@Tad was significantly increased, and the area under the curve was increased by about 7.88 times. In the pharmacodynamic study, iRGD-lipo@Tad achieved a continuous and efficient improvement of placental blood perfusion. This was achieved by decreasing the ratio of plasma soluble fms-like tyrosine kinase to placental growth factor and increasing the levels of cyclic guanosine monophosphate and nitric oxide. Consequently, iRGD-lipo@Tad resulted in a significant increase in embryo weight and a reduction in the miscarriage rate of N-Nitro-L-arginine methyl ester-induced IUGR pregnant mice without detectable toxicity. In summary, the nanotechnology-assisted therapy strategy presented here not only overcomes the limitations of tadalafil in the clinical treatment of IUGR but also offers new avenues to address the treatment of other placenta-originated diseases.

Carbon monoxide affects early cardiac development in an avian model.

Carbon monoxide (CO) is a toxic gas that can be lethal in large doses and may also cause physiological damage in lower doses. Epidemiological studies suggest that CO in lower doses over time may impact on embryo development, in particular cardiac development, however other studies have not observed this association. Here, we exposed chick embryos in ovo to CO at three different concentrations (3, 9, 18 ppm) plus air control (4 protocols in total) for the first 9 days of development, at which point we assessed egg and embryo weight, ankle length, developmental stage, heart weight, ventricular wall thickness, ventricular-septal thickness and atrial wall thickness. We found that heart weight was reduced for the low and moderate exposures compared to air, that atrial wall and ventricular wall thickness was increased for the moderate and high exposures compared to air and that ventricular septal thickness was increased for low, moderate and high exposures compared to air. Ventricular wall thickness was also significantly positively correlated with absolute CO exposures across all protocols. This intervention study thus suggests that CO even at very low levels may have a significant impact on cardiac development.

Impact of Human Adenovirus 36 on Embryonated Chicken Eggs: Insights into Growth Mechanisms.

Human adenovirus 36 (HAdV-D36) is presently the sole virus identified to be associated with an elevated risk of obesity in both humans and animals. However, its impact on embryonated chicken eggs (ECEs) remains unexplored. This study endeavoured to examine the influence of HAdV-D36 on embryonic development by utilizing embryonated chicken eggs as a dynamic model. To simulate various infection routes, the allantoic cavity and the yolk sac of ECEs were inoculated with HAdV-D36. Subsequently, embryos from both the experimental (inoculated with virus) and control (inoculated with PBS) groups were weighed and subjected to daily histological examination. The daily embryo weights were assessed and compared between groups using the Shapiro-Wilk test. Histopathological changes in tissues were examined and compared between the tested and control groups to ascertain physiological alterations induced by the virus. Our study confirmed a significant increase in the body weight of ECEs. However, this phenomenon was not attributable to adipose tissue development; rather, it was characterized by an augmented number of cells in all observed tissues compared to control subjects. We posit that HAdV-D36 may impact developing organisms through mechanisms other than enhanced adipose tissue development. Specifically, our findings indicate an increased number of cells in all tissues, a phenomenon that occurs through an as-yet-unexplored pathway.

Adiponectin and visfatin expression profile in extra-embryonic annexes and role during embryo development in layer and broiler chickens

Some evidence showed differences between layer and broiler embryo development. We recently showed that two adipokines, adiponectin and visfatin are expressed in the extra embryonic membranes and fluids. However, their role in the embryo development is unknown. Thus, our objectives were 1. to compare the expression of AdipoQ and its receptors AdipoR1 and AdipoR2 and visfatin in extra-embryonic annexes in broiler and layer breeders during the embryo development and 2. to investigate the role of two adipokines in embryo development in both broiler and layer breed after in ovo injection of blocking antibodies against chicken adiponectin or visfatin. We found that adiponectin, AdipoR1, AdipoR2 and visfatin were mainly more expressed in the allantoic that in amniotic membranes. In addition, these expressions increased according the stage of embryo development. We observed a higher expression in layer than in broiler of AdipoQ in allantoic membranes at ED14 and ED18, of AdipoR1 and AdipoR2 in both allantoic and amniotic membranes at ED7 and ED14 and of visfatin only in allantoic membrane from ED7 to ED18. AdipoQ and visfatin were absent in amniotic fluid at ED7 but present at ED14 or ED18 where higher concentrations were detected in layer than in broiler. Interestingly, we showed a strong positive correlation between Adipo and visfatin concentration in amniotic fluid and the body weight of embryo in both breeds. However, after in ovo injection of Adipo antibodies we did not observe any effect on the embryo mortality whereas injection of visfatin antibodies increased in a dose dependent manner the embryo mortality in both breeds. Taken together, Adipo and visfatin are higher expressed in layer than broiler in extra-embryonic membranes and amniotic fluid. Our data suggest also that visfatin could be a main regulator of embryo development.

Effect of Curcuma longa alcoholic extract on Mice Embryos Development

This study focused on determining the effect of the alcoholic extract of curcuma longa on mouse embryo, twenty adult white Swiss mice used in this study. Experimental animals were divided into four groups with six mice for each group. For mating occur, 3 females and male were placed in cage. It was confirmed that the mating took place when examining the next morning and watching the vaginal plug, and considered that the day after mating is the first day of the pregnancy. We prepared three subs lethal concentrations of curcuma longa were applied to Experimental groups except control group, 10 mg/kg, 50 mg kg/ and 100 mg/kg. From 7-14 of pregnancy we were given the pregnant mice 0.1 ml per 10 gm body weight. After 18 of pregnancy we extracted the embryo to study the effect of alcoholic extract and weights and lengths embryos and study the embryo malformation, as well as a study the weight of mice before and after experiment. This study shows a significant (P&lt;0.01) decrease in the body weight of mice that treated with three concentrations (10 -50 -100) mg/kg compared with control group, but there is not significant different in lengths and weights of mice embryos .concentrations 50 mg/kg and 100 mg/kg show much malformation induced in the mice compared with embryos of control group, include cleft lips, hemorrhage, and Micromelia , as well as we observed absorbed embryos in 50 mg/kg and 100mg/kg . This study concluded that alcoholic extract of (curcuma longa) has teratogenic effects on embryos of mice.&#x0D; Keywords: curcuma longa, embryonic malformation, Micromelia , teratogenic effects.

Impact of storage duration and short periods of incubation during egg storage on embryonic development and hatching traits of hybrid chicken strain

The study assessed chicken egg storage duration and short periods of incubation during egg storage (SPIDES) on embryo development. Eggs samples from 310 eggs from 72-week-old hybrid layer breeder flocks were divided and stored at 16 °C and 75% RH for 1, 4, 7, 10, 14, 17, and 21 days (D). Some eggs were also divided between 14D and 21D alone and stored. One subgroup received heat application for 6 h during storage resulting in 0-h × 14D (control); 0-h × 21D (control); 6-h × 14D (SPIDES) and 6-h × 21D (SPIDES). Storage durations between 1D and 21D did not influence most egg quality parameters. The interaction of storage duration and SPIDES affected initial, final egg weights and egg weight loss. SPIDES eggs recorded heavier wet embryos than non-SPIDES eggs at embryonic days (ED) 4, ED7, ED11, and dry embryo weight at ED11. SPIDES resulted in longer embryos at ED4 and ED11. In 21D stored eggs hatchability was higher in SPIDES compared to non-SPIDES treatments (66.7 vs. 48.3%). The reverse effect of SPIDES occurred in 14D stored eggs. In conclusion, the study shows positive effect of SPIDES in longer stored eggs and could be due to a positive impact on embryo.

Synthesis and evaluation of biological effects of modified graphene oxide nanoparticles containing Lawson (Henna extract) on gastric cancer cells.

Targeted Graphene Oxide (GO) nanoparticles can play an important role in the treatment of cancer by increasing cancer cell targeting. This study was conducted to synthesize GO nanoparticles functionalized with chitosan-folate (CS-FA) to deliver a natural product Lawsone (LA) for cancer treatment. After characterization of the LA-GO-CS-FA, antioxidant activities of the nanoparticles were investigated by ABTS, DPPH, and FRAP tests. CAM assay was used to study the effect of nanoparticles on angiogenesis. The expression level of inflammatory and angiogenic genes in cells treated with nanoparticles was evaluated by real-time PCR. The findings demonstrated the formation of nanoparticles with a size of 113.3nm, a PDI of 0.31, and a surface charge of + 11.07mV. The percentages of encapsulation efficiency were reported at 93%. Gastric cancer cells were reported as the most sensitive to treatment compared to the control, and the gastric cancer cells were used to study gene expression changes. The anti-angiogenic effects of nanoparticles were confirmed by reducing the average number and length of blood vessels and reducing the height and weight of embryos in the CAM assay. The reducing the expression of genes involved in angiogenesis in real-time PCR was demonstrated. Nanoparticles displayed high antioxidant properties by inhibiting DPPH and ABTS radicals and reducing iron ions in the FRAP method. The reduction of pro-inflammatory genes in AGS cells which were treated with nanoparticles indicates the anti-inflammatory properties of nanoparticles. This study showed the efficacy of nanoparticles in inhibiting gastric cancer cells by relying on inhibiting angiogenesis.

Research Note: In ovo and in-feed probiotic supplementation improves layer embryo and pullet growth

Probiotics are widely used as feed supplements in the poultry industry to promote growth and performance in chickens. Specifically, this supplementation starts around the time of lay and continues through the production cycle in laying hens. However, the embryonic period is critical to the growth and development of metabolically active organs thereby influencing subsequent health and productivity in adult birds. Therefore, the present study investigated the potential use of probiotics to promote embryonic growth in layers. Further, a pilot grow-out study was conducted to evaluate the effect of in ovo and in-feed probiotic application on pullet growth. For the study, fertile White Leghorn eggs were sprayed with phosphate buffered saline (control, CON) or probiotic cocktail (in ovo only, IO; Lactobacillus paracasei DUP 13076 and L. rhamnosus NRRL B 442) prior to and during incubation. The embryos were sacrificed on d 7, 10, 14, and 18 of incubation for embryo morphometry. On d 18, remaining eggs were set in the hatcher to assess hatchability and hatchling morphometry. For the pullet trial, hatchlings were raised on feed with or without probiotics until wk 5. Pullets were sacrificed weekly, and morphometric parameters were recorded. Results of our study demonstrate that in ovo probiotic application significantly improved relative embryo weight, crown-rump length, hatchability, and hatchling weight when compared to the control (P < 0.05). Further, this enhanced embryonic development was associated with a concomitant increase in posthatch growth. Specifically, pullets raised from probiotic-sprayed eggs had significantly improved crown-rump length, tibial length, tibial bone weight, and body weight when compared to the control (P < 0.05). Moreover, among the different treatment schemes employed in this study [CON (no probiotics), in-feed only (IF), IO only, and in ovo and in-feed probiotic supplementation (IOIF)], sustained probiotic supplementation (IOIF) was found to be the most effective in promoting growth. Therefore, in ovo and in-feed probiotic supplementation could be employed to promote embryo and pullet growth to support subsequent performance in layers.

Egg mineral levels, hatching results and bone properties in embryos and chicks obtained from broiler breeders at different ages.

It is inevitable to obtain chicks from breeders of different ages in broiler fattening. However, breeder age has an effect on eggs. There is no detailed study on the changes in mineral levels in eggs and bones of these effects. Therefore, the aim of this study examines how mineral levels of shell, albumen, egg yolk, and bone, embryonic development and bone characteristics change according to the breeder age. A total of 370 fresh hatching eggs were obtained from Ross-308 breeders at 2 different ages (25 to 60-week-old). Egg, embryo, yolk sac and chick characteristics were determined. Morphometric parameters of bones of embryos at 19 days of age and chicks at hatch were examined. Mineral levels of albumen, yolk, shell and bones of embryo and chick were determined. Egg weight, embryo weight, embryo length, chick weight and chick length were higher in old breeder than those of younger one. Femur length, width and Seedor index of tibia, weight, width and Seedor index of metatarsus were found to be high in embryos at 19 days of old breeder than young breeder. There were differences in relative femur weight, tibia length and relative metatarsus weight of chicks at hatch. The mineral levels were high in the femur, tibia and metatarsus of embryos and chicks, in parts of the egg of old breeder. K, Ca, Mg, P, Fe and Mn levels in the eggshell were decreased during incubation. Ca and Mg levels were decreased in the yolk during incubation. K, Ca, Mg, P, Fe, Zn and Mn levels were found to be high in chick bones than those of bones of embryos at 19 days of age. The total embryonic death was found to be high in the fertile eggs of younger breeder group. No differences were observed in fertility, hatchability of fertile and set eggs according to the age of breeders. These data may be useful in designing in-ovo injections, formulating diets of broiler breeders and broilers and in preventing bone diseases in broilers.

Effect of Embryo Weight and Seed Priming on Seedling Characteristics of Different Maize Cultvars

An laboratory experiment was carried out during the year 2020 at the Seed Technology Laboratory of the Crop Department - College of Agricultural Engineering Sciences - University of Baghdad in order to know the effect of seed priming and size of their embryos of different cultivars of maize on the seedling characteristics. The experiment was carried out in a completely randomized design with four replicates. The first factor included three synthetic cultivars of maize (Al-Maha, Baghdad 3 and Al-Fajr) and Al-Nahrain hybrid. For the second factor, it included the priming of seeds with ASA, CK, GA3, KCL, H2O2, DW and DS, in addition to the dry seeds (without priming). The seeds were soaked with solutions of these substances for 24 hours and six characteristics related to the germination process were estimated, which were the weight of the seed after stimulation, the weight of the embryo, the time of germination, the length of the radicle and shoot, and its dry weight. The results showed that the seed priming with H2O2 was significantly superior, as it gave the highest average of seed weight, lowest average germination time, longest average radiclelength and highest seedling dry weight (0.336 g, 3.12 days, 15.08 cm and 0.215 g, respectively). The treatment of seeds with ascorbic was superior in giving the highest average weight of the embryo that reached 0.049 gm, the treatment of stimulating the seeds with gibberellin was superior in giving the highest mean of the coleoptile length of 10.79 cm. Al-Maha cultivar excelled in giving it the highest average seed weight, embryo weight, radiclelength and dry weight, as it reached 0.335 g, 0.050 g, 14.42 cm and 0.194 g, respectively. Whereas, the Fajr cultivar was significantly superior by giving the highest average coleoptile length of 9.37 cm. The interaction effect between the study factors was significant in most of the studied traits.