Relative Embryo Weight Research Articles

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.

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.

Modulatory Effects of Arctostaphylos uva-urs Extract In Ovo Injected into Broiler Embryos Contaminated by Aflatoxin B1.

Simple SummaryThe poultry industry is of great importance worldwide; however, mycotoxins contamination, especially aflatoxins, has harmful effects on poultry production. Therefore, finding out new strategies to alleviate aflatoxin contamination is an important trend of research. In our study, we focused on the in ovo injection techniques using the methanolic extract of brae beery “Arctostaphylos uva-ursi” (Ar. uu) to modulate the aflatoxinB1 (AFB1) adverse effects in broiler embryos. As a result, 0.1 g of Ar. uu in ovo injection alleviates AFB1 embryotoxicity by enhancing chicks’ physiological responses and optimizing antioxidant status, which is reflected in a low mortality rate and heavier relative weight of the hatched chicks. The in ovo injection of AFB1 + Ar. uu significantly declined AFB1-induced toxicity in embryos and increased broiler chicks embryo’s survival by 62.5%, increased relative embryo weight by ∼65.25% compared to eggs injected with AFB1 alone. Regarding immune responses, the in ovo injection of Ar. uu enhances the embryo’s humoral immune responses and regulates oxidative stress biomarkers. In general, the in ovo injection of Ar. uu modulates broiler chicks’ embryotoxicity caused by AFB1.In ovo injection of nutrients can modulate the embryo’s physiological responses against aflatoxin B1 (AFB1) embryotoxicity. This hypothesis was tested using in ovo injection of Arctostaphylos uva-ursi (Ar. uu.) methanolic extract. The total polyphenols, total flavonoids, total antioxidant capacity, and GC-MS analysis were all assessed in the Ar. uu. methanolic extract. A total of 180 ten-day-old embryonated eggs were distributed into six groups of 30 replicates each. The first group was used as a control (non-injected), and the second, third, fourth, fifth, and sixth groups were injected with 10 µ double-distilled water (DDW), 500 µL methanol, 0.01 g Ar. uu./500 µL methanol, 50 ng AFB1/10 µL DDW, and 50 ng AFB1 in 10 µ DDW + 0.01 g Ar. uu./500 µL methanol, respectively. The relative embryo weight, residual yolk sac weight, tibia length and weight, and survival were recorded. Total and differential leukocytes, oxidative stress, and humoral immune responses were observed. The residual yolk sac was lower (p < 0.05) in the Ar. uu. group than other groups. The embryonic growth (tibia weight and length) was enhanced in AFB1 + Ar. uu.-injected embryos compared with those injected with AFB1 alone. In conclusion, in ovo injection of Arctostaphylos uva-ursi could modulate AFB1-induced toxicity in chicken embryos.

Slightly acidic electrolyzed water as an alternative disinfection technique for hatching eggs

Conventional chemical disinfectants used for egg disinfection could result in toxic residue and endanger hatchability, chick quality, and pullet growth performance. Slightly acidic electrolyzed water (SAEW) is known as a novel disinfectant for egg sterilization due to its high efficiency and no residue. In this study, a comprehensive assessment of slightly acidic electrolyzed water and benzalkonium bromide solution (BBS) used in the disinfection channel was conducted to assess the microbial count, eggshell quality, and hatchability concomitantly. The results show that the sterilization efficiency of SAEW increased with an increase in available chlorine concentration (ACC), spraying volume, and sterilization duration. SAEW with an ACC of 150 mg/L and 10,000 mg/L benzalkonium bromide solution had the same sterilization rates of approximately 86.2% at a spraying volume of 0.5 mL/egg and sterilization duration of 180 s. Neither had significant effect on eggshell strength or thickness. The eggshell cuticle quality in the benzalkonium bromide group was significantly higher than the control group (no disinfection) and the 150 mg/L SAEW group. The embryo weight, relative embryo weight, hatchability, and embryonic mortality in the SAEW group had no significant differences of those in the benzalkonium bromide group. SAEW should be more popular because of its simple preparation, low cost, and no residue. Our results indicate SAEW is an alternative disinfectant for the sterilization of hatching eggs instead of conventional chemical disinfectants, such as benzalkonium bromide, and give a recommendation is using SAEW as a disinfectant with 150 mg/L ACC, 0.5 mL/egg spray volume, and disinfection for 180 s in the novel disinfection channel.

The effects of in ovo administered bisphenol A on tibial growth plate histology in chicken

The aim of this study was to determine of the effects of in ovo administered BPA on embryonic development of the tibial growth plate using histological methods in chickens. Three hundred and ten fertile eggs of Isa Brown laying parent stock were divided into five groups as untreated control, vehicle-injected control, 50, 100, and 250 μg/egg BPA. At the 13th, 18th, and 21st days of incubation, eggs were randomly opened from each group until 10 live embryos were obtained. Embryos were weighed and crown-rump length was measured. Tibial tissue samples were taken from embryos. Tibia weight, relative tibia weight and tibia length were determined. Tissue samples were fixed in 10% buffered formalin solution. Sections were stained with Safranin O staining methods and zones in the growth plate were measured. Also, proliferating cell nuclear antigen (PCNA) was stained immunohistochemically. The mortality in the BPA treated groups was higher than untreated control group. The results have revealed that mean relative embryo weights, crown-rump length, mean tibia weight, relative tibia weight, and tibia length of BPA treated groups were significantly lower when compared to the untreated control and vehicle-injected control groups. Also, proliferative zone get significantly narrowed, whereas the transitional and hypertrophic zone thickened and PCNA positive chondrocytes increased in growth plate of BPA treated groups. These results have suggested that developmental exposure to BPA adversely affected development of the tibial growth plate.

Embryonic thermal manipulation of Japanese quail: effects on embryonic development, hatchability, and post-hatch performance.

Embryonic thermal manipulation led to several modifications in molecular, physiological, and biochemical parameters which affect pre- and post-hatch growth performance. The current study aims to elucidate the onset and long-term effects of intermittent thermal manipulations (TM) during two-time windows, early/late, of embryogenesis in Japanese quail (Coturnix japonica) on embryonic development, hatchability, muscle histogenesis, and post-hatch growth performance. Four groups were created; quail eggs in the control group were incubated at 37.7 °C and relative humidity (RH) 55%. Three thermally treated groups were incubated intermittently at 41 °C and 65% RH intermittently (3 h/day): early embryogenesis group (EE) was thermally treated during embryonic days (ED) 6-8, late embryogenesis group (LE) was thermally treated during (ED12-ED14), and early and late embryogenesis group (EL) was thermally manipulated in both time windows. Relative embryo weights in EL and EE were significantly lighter than those in LE and Ctrl groups. The hatched chicks were reared under optimal managemental conditions (three replicates per treatment). Average daily feed intake was recorded, and feed conversion ratio (FCR) was calculated. Histological and quantitative analyses of muscle fibers were performed. The results revealed that TM led to significant hypertrophy of quail breast muscle in (EE). Intermittent short-term (3-6 h) thermal manipulation (39-40 °C) protocols during early embryogenesis (ED6-ED8) could be recommended to enhance muscle mass growth and breast muscle yield in the Japanese quail.

The Embryotoxic Effects of in Ovo Administered Sunset Yellow FCF in Chick Embryos.

Sunset yellow (SY) at prescribed concentrations has been approved by regulatory authorities in several countries as an additive dye in the food, beverage, cosmetic, and pharmaceutical industries. However, there are some reports that it may cause several health problems. The aim of this study is to evaluate embryotoxic effects of SY on liver and kidney in chick embryos. Babcock white Leghorn eggs were randomly divided into four groups. Non-treated eggs served as control group. The eggs in groups SY200, SY1000, and SY2000 were treated with a single injection of 200, 1000, and 2000 ng SY into the air sac just before incubation. The developmental stages of embryos were determined on the 10th, 13th, 16th, and 21st days of incubation. Samples of the liver and kidney were taken and routine histological procedures were performed. The highest relative embryo weight was seen in all SY treated groups on the 16th day of incubation. Necrosis of some hepatocytes and cytoplasmic degenerations were observed in all SY groups in the liver. There were degenerated or destructed renal cortex structures and necrosis in the kidney. The cell’s nuclear areas and diameters of renal cortex structures were different in all SY groups compared to the control group (p < 0.05). It was concluded that in ovo administered SY has many unfavorable effects on liver and kidney in chick embryos. The results obtained in this study suggest that it may be advisable to re-assess safety levels of SY in many industries.

Differential physiological response of slow- and fast-growing broiler lines to hypoxic conditions during chorioallantoic membrane development

Ambient conditions during chicken embryogenesis, such as insufficient oxygen or changes in temperature, are expected to cause permanent phenotypic changes and affect their posthatch performance. Decades of genetic selection for high growth rate resulted with various physiological and morphological changes that can affect the broiler fitness under environmental stress. To evaluate the selection effect on responses to environmental challenge during embryonic development, and the long-term implications, we have used a unique genetic line, that was not selected for over 30 yr (since 1986), as control for the modern commercial genetic line. At embryonic day 5 (E5), broiler embryos from these 2 genetic lines were divided into 2 treatments: 1) control; 2) 15% O2 concentration for 12 h/day from E5 through E12 the embryonic period of chorioallantoic membrane formation. Embryos and hatched chicks were characterized for physiological and morphological parameters. Significant differences in relative embryo weight and yolk consumption were found between the 2 lines. The modern line was characterized by a higher metabolic rate and rapid growth, supported by higher hemoglobin levels and hematocrit concentrations, whereas the 1986 line had slower metabolism, lower levels of hematocrit and hemoglobin, higher oxygen volume per 1 g of embryonic tissue indicating higher oxygen availability. Both lines exhibited changes in heart rate, and blood parameters corresponding to cardiovascular system adaptation after hypoxic exposure, seemingly implemented to increase oxygen-carrying capacity to the embryo tissues. Our finding stand in agreement that the genetic selection for high growth rate that led to higher metabolism without a fit of the cardiovascular system, increased the imbalance between oxygen supply and demand.

Effects of dietary supplementation with vitamin E, selenium yeast or both on egg incubation response, embryonic development, keet quality, and posthatch growth of helmeted guinea fowl breeders.

This study investigated the effects of dietary supplementation with vitamin E (vit. E), selenium yeast (Se yeast), or both on egg incubation response, embryonic development, keet quality, and posthatch growth of helmeted guinea fowls. Two hundred and forty 24-week old helmeted guinea fowl hens (average weight 1.75+0.22kg) and cocks (average weight 2.15+0.20kg) were assigned into 24 pens; each pen housed 10 hens and 2 cocks. There were four dietary treatments consisting of a basal diet (control), basal diet supplemented with vit. E (30IU/kg), Se yeast (0.3mg/kg Se), or both. Six pens were assigned to each treatment. Egg incubation response were estimated using 504 settable eggs sampled from each treatment collected during 15 to 17weeks in lay. A total of 72 fertile eggs sampled from each treatment were used for the estimation of embryonic development. Quality of day-old keets hatched was scored based on physical conditions, while posthatch growth was measured for 21days. Guinea fowl breeders fed diet supplemented with both vit. E and Se yeast produced the highest (P < 0.05) number of fertile eggs, percentage fertility, number of hatchlings, hatchability of total eggs, and hatchability of fertile eggs. Supplementation with vit. E + Se yeast resulted in the heaviest (P < 0.05) embryo weight, relative embryo weight, least (P <0.05) yolk sac weight, and relative yolk sac weight on 25days of incubation. Hatchlings from breeders fed diet supplemented with Se yeast and vit. E + Se yeast showed normal swallowed yolk. Supplementation of maternal diet with vit. E, Se yeast, and vit. E + Se yeast resulted in improved (P < 0.05) feed conversion ratio of subsequent hatchlings during 1 to 7-day posthatch growth. It can be concluded that dietary supplementation of vit. E + Se yeast in guinea fowl breeders resulted in improved egg fertility, hatchability, heavier embryo weights, hatchlings of good quality, and improved posthatch growth during the first 7days.

Phytochemicals reduce aflatoxin-induced toxicity in chicken embryos

Aflatoxins (AF) are toxic metabolites produced by molds, Aspergillus flavus and Aspergillus parasiticus, which frequently contaminate poultry feed ingredients. Ingestion of AF-contaminated feed by chickens leads to deleterious effects, including decreased bird performance and reduced egg production. Moreover, AF residues in fertilized eggs result in huge economic losses by decreasing embryo viability and hatchability. This study investigated the efficacy of 2 generally recognized as safe phytochemicals, namely carvacrol (CR) and trans-cinnamaldehyde (TC), in protecting chicken embryos from AF-induced toxicity. Day-old embryonated eggs were injected with 50ng or 75ng AF with or without 0.1% CR or TC, followed by incubation in an incubator for 18 d. Relative embryo weight, yolk sac weight, tibia weight, tibia length, and mortality were recorded on d 18 of incubation. The effect of phytochemicals and methanol (diluent) on embryo viability was also determined. Each experiment had ten treatments with 15 eggs/treatment (n = 150 eggs/experiment) and each experiment was replicated 3 times. Both phytochemicals significantly decreased AF-induced toxicity in chicken embryos. At 75ng of AF/egg, CR and TC increased the survival of chicken embryo by ∼55%. Moreover, CR and TC increased relative embryo weight by ∼3.3% and 17% when compared to eggs injected with 50ng or 75ng AF, respectively. The growth of embryos (tibia length and weight) was improved in phytochemical-treated embryos compared to those injected with AF alone (P <0.05). Phytochemical and methanol treatments did not adversely affect embryo survival, and other measured parameters as compared to the negative control (P >0.05). Results from this study demonstrate that CR and TC could reduce AF-induced toxicity in chicken embryos; however, additional studies are warranted to delineate the mechanistic basis behind this effect.

The effects of different eggshell temperatures on embryonic development, hatchability, chick quality, and first-week broiler performance

The aim of the current study was to determine the effects of different eggshell temperatures (EST) during 10 to 18 d of incubation on embryonic development, hatchability, chick quality, first-week broiler performance. The EST were maintained within the temperature ranges of 33.3 to 36.7, 37.8 to 38.2, and 38.9 to 40.0°C for the low, control, and high EST treatments, respectively. From d 15 to 18, embryo weight and relative embryo weight were found to be similar in the low and high EST groups. Salable chicks and hatchability of total eggs was found to be higher in the control EST group. Between d 10 and 17 of incubation, embryonic mortality in low, control, and high EST groups was determined to be 1.6, 0.8, and 2.0%, respectively. From d 18 to hatch, embryonic mortality and rate of dead and cull chicks were found to be significantly different. Hatching in high EST group was completed 26 h early, although hatching in low EST group was completed 10 h later than the control EST group. On the of hatching day, chick weight and length were found to be 39.5, 41.0, and 42.5 g, and 18.5, 21.4, and 19.1 cm in low, control, and high EST groups, respectively. The highest residual yolk sac weight and relative residual yolk sac weight were observed in high EST group as 7.7 g and 18.7%. Yolk-free chick weight and relative yolk-free chick weight were highest in the control EST group. At 1 wk of age, the BW in low, control, and high EST groups were determined as 131.1, 140.0, and 140.8 g, respectively. No significant difference was found for feed intake and feed conversion among treatments for wk 1. The mortality during the first week did not differ among groups; however, a higher mortality rate was observed numerically in the high EST group. In conclusion, embryo development, incubation parameters, chick quality, and the first week performance are affected by small changes in the EST.

Estimation of Weight and Lipid Composition in Preimplantation Embryos from Jersey and Beef Breeds of Cattle

Cryopreservation is the main functional means for storage of excess embryos produced from artificial reproductive technologies; the process assumes embryos chemical nature is highly conserved across embryos of the same species. However, in practice there appears to be a high degree of variability in embryo tolerance to cryopreservation, suggesting potential differences in embryo chemistry. The objective of the current study was to develop reproducible means of estimating relative embryos weight and associating those weights to lipid content. Relative embryo weights of frozen/thawed in-vivo Jersey and Crossbred beef breed embryos were determined using a modified specific gravity chamber. Embryo weights were then correlated with lipid content. Results suggest that beef cattle embryos are 20% - 27% heavier than Jersey embryos (P

Interaction between ascites susceptibility and CO during the second half of incubation of two broiler lines. Effect on embryonic development and hatching process

1. Because CO2 during the second half of incubation is known to influence air cell and blood gases, and embryo development, it is postulated that post-hatch development and ascites sensitivity could also be influenced. 2. An ascites susceptible (A) and an ascites resistant (E) broiler line were incubated under standard incubation or high CO2 conditions (up to 4%) from embryonic day (ED) 10 onwards. The embryonic development and the hatching process of these two lines were compared when incubated under standard or high CO2 conditions from over the second half of incubation. 3. The A line, selected for high post-hatch growth rate, exhibited a higher relative embryo weight from ED10 until ED16, which was supported by a higher air cell pCO2, lower air cell pO2, higher corticosterone and thyroid hormones and earlier hatching time. 4. Incubation under high CO2 increased air cell pCO2, retarded yolk consumption, and decreased glycogen concentration in the liver at hatch. Hatchability decreased in both lines when incubated under high CO2, due to an increased late mortality of embryos that died before IP. 5. These results suggest that the development and metabolism of CO2-incubated embryos differ from control incubated embryos.

Influences of Hypoxia on Hatching Performance in Chickens with Different Genetic Adaptation to High Altitude

The experiments were conducted to assess how hatching performance is affected by chicken breeds and environment of high altitude and to analyze the vital factor of the low hatchability at a 2,900-m altitude. Eggs of Tibetan and Dwarf chickens were incubated at conditions of normobaric normoxia, normobaric hypoxia, hypobaric hypoxia, and supplemental O2 at high altitude (hypobaric normoxia) during the whole incubation or at 0 to 7, 8 to 14, and 15 to 22 d of incubation, respectively. The results showed that the Tibetan chickens had greater hatchability (79.72%), lower water loss (12.90%), greater relative embryo weight (38.08%), and relative chick weight (68.41%) compared with the Dwarf chickens (31.69, 15.79, 30.71, and 65.21%, respectively) when both of them were incubated at a 2,900-m altitude. The hatchability was 71.60% in Tibetan chicken and 36.23% in Dwarf chicken under the normobaric hypoxia condition. The hatchability of chicken was efficiently increased with supplemental O2. The previous results indicated that the O2 deficit is the main factor resulting in the low hatchability and the poor chick quality of the lowland chicken breed when incubated at a 2,900-m altitude. Breeding chickens for adaptability to hypoxia and supplemental O2 is a good way to improve the hatchability and chick quality at that altitude.