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Changes in physicochemical parameters of duck eggs and extra-embryonic structures during incubation.Animal : An International Journal of... Dec 2023Duckling embryogenesis should be deepened due to the hatching technology and its modification possibilities. Many changes occur in incubated eggs, which expose the...
Duckling embryogenesis should be deepened due to the hatching technology and its modification possibilities. Many changes occur in incubated eggs, which expose the embryo to hazards. The study aimed to analyse the physicochemical properties of eggshell, yolk, thick albumen (TA), and amniotic fluid (AF) of incubated hatching eggs from 52-week-old Cherry Valley ducks. The morphological features of 18 fresh eggs were analysed. Over 28 days, a total of 800 eggs underwent incubation. Eggshell surface temperature and egg weight loss were measured on days 1, 4, 7, 10, 14, 18, 21, and 25. Eggshell, TA, AF, and yolk were collected from eggs at incubation days 1-21 (every week). TA was collected on days 0, 1, and 7, while AF on days 7, 14, and 21. The analysis covered a range of physicochemical parameters. Eggshell thickness decreased with incubation, reaching its lowest point posthatch (P < 0.001). The highest pH for TA was recorded on day 1, while the lowest was on day 7 when comparing days 0, 1, and 7 (P < 0.001). TA pH was consistently higher than in AF (P < 0.001). However, the pH of TA was the highest on day 1 and the lowest on day 7 (P < 0.001). Yolk pH increased from days 1 to 21 (P < 0.001). There was also a noticeable in egg weight loss (0.34% daily) (P < 0.001). Vitelline membrane strength decreased from day 0 to day 1 (P < 0.001). Lysozyme activity in thick albumen on day 7 was higher than on days 0 and 1 (P < 0.001). Lysozyme activity in AF was higher on day 21 than days 7 and 14 (P < 0.001). TA viscosity was highest on day 0 and lowest on day 1, compared to other days (P < 0.001). AF viscosity and CP content exhibited an increase on day 21 as compared to days 7 and 14 (P < 0.001). The CP content in TA was notably higher on day 7 than on days 0 and 1 (P < 0.001). Polyunsaturated fatty acids declined, while monounsaturated and transfatty acids increased (P < 0.001). Viscosity and lysozyme activity increased on day 7 in TA and day 21 in AF. TA and the amniotic cavity appeared to facilitate the transfer of substances, particularly CP. Viscosity could be an indicator for optimising incubation conditions, as incorrect changes can affect embryo mortality. The results showed the different utilisation of nutrients, such as fatty acids. It could support research on the in-ovo administration of various substances.
Topics: Animals; Ducks; Muramidase; Ovum; Egg Shell; Weight Loss; Eggs; Chickens
PubMed: 37981451
DOI: 10.1016/j.animal.2023.101024 -
Brazilian Journal of Biology = Revista... 2023Specimens of Cnemidocarpa amphora were collected monthly from the Arabian Gulf from September 2017 to August 2018. Parts of their gonads were prepared for histological...
Gonadal proliferation and reproductive cycle of the exotic sea squirt Cnemidocarpa amphora () (Pleurogona, Styelidae) sampled for the first time from the northern coast of Arabian Gulf in Saudi Arabia.
Specimens of Cnemidocarpa amphora were collected monthly from the Arabian Gulf from September 2017 to August 2018. Parts of their gonads were prepared for histological testing. The gonads' diameters varied by month. Each gonad contained many ovarian follicles with different morphologies and was surrounded by several testicular follicles. The ovarian and testicular follicles were separate, although the latter were always present near the former. Repeated measures ANOVA tests were conducted to investigate monthly changes in oocyte stages. In squirts measuring 12-13 cm in length, the gonads measured 30-50 mm from July to August; 20-25 mm from September to October; 15-20 mm from November to February; and 25-40 mm from March to June. Oogonia budded from the germinal epithelium with diameters of 20-30 µm. Previtellogenic oocytes measuring 70-120 µm were characterized by the deposition of small granules of protein around the nucleus, a continuous layer of follicular cuboidal epithelium, and scattered vacuoles in the ooplasm. The measurement of gonads and oocyte diameters were performed by image analysis (Image scope 2.3, Image Line, Inc.) and stage micrometer. The vitellogenic oocytes measured 130-220 µm and the follicular epithelium consisted of flattened and cuboidal layers. Beneath the vitelline membrane, scattered test cells appeared in the ooplasm and different granules of protein and MPS were deposited in the ooplasm. In the later phase, lipid droplets began to appear in the ooplasm. Yolk bodies formed after the impregnation of various granules together and the oocyte was ready to be shed. Before spawning, a yolk membrane appeared above the ooplasm. Post-vitellogenic oocytes, in which the homogeneity of ooplasm was restored, underwent gradual lysis and entered the atretic phase. Different stages of sperm development were present year-round in different follicles of the same squirt; hence, the testes were always mature.
Topics: Animals; Female; Male; Urochordata; Saudi Arabia; Semen; Oocytes; Ovary; Cell Proliferation
PubMed: 37970899
DOI: 10.1590/1519-6984.273666 -
Scientific Reports Nov 2023The specific functions and essentiality of type II vitellogenin (Vtg2) in early zebrafish development were investigated in this study. A vtg2-mutant zebrafish line was...
Genomic disturbance of vitellogenin 2 (vtg2) leads to vitellin membrane deficiencies and significant mortalities at early stages of embryonic development in zebrafish (Danio rerio).
The specific functions and essentiality of type II vitellogenin (Vtg2) in early zebrafish development were investigated in this study. A vtg2-mutant zebrafish line was produced and effects of genomic disturbance were observed in F2 females and F3 offspring. No change in vtg2 transcript has been detected, however, Vtg2 abundance in F2 female liver was 5×, and in 1 hpf F3 vtg2-mutant embryos was 3.8× less than Wt (p < 0.05). Fecundity was unaffected while fertilization rate was more than halved in F2 vtg2-mutant females (p < 0.05). Hatching rate was significantly higher in F3 vtg2-mutant embryos in comparison to Wt embryos. Survival rate declined drastically to 29% and 18% at 24 hpf and 20 dpf, respectively, in F3 vtg2-mutant embryos. The introduced mutation caused vitelline membrane deficiencies, significant mortalities at early embryonic stages, and morphological abnormalities in the surviving F3 vtg2-mutant larvae. Overrepresentation of histones, zona pellucida proteins, lectins, and protein degradation related proteins in F3 vtg2-mutant embryos provide evidence to impaired mechanisms involved in vitellin membrane formation. Overall findings imply a potential function of Vtg2 in acquisition of vitellin membrane integrity, among other reproductive functions, and therefore, its essentiality in early zebrafish embryo development.
Topics: Animals; Female; Embryo, Nonmammalian; Embryonic Development; Genomics; Larva; Vitellins; Vitellogenins; Zebrafish
PubMed: 37914813
DOI: 10.1038/s41598-023-46148-2 -
Poultry Science Dec 2023The study aimed to analyze the hatching egg and physiochemical features of eggshells, thick albumen, amniotic fluid, and yolk during the incubation of Ross 308 chicken...
The study aimed to analyze the hatching egg and physiochemical features of eggshells, thick albumen, amniotic fluid, and yolk during the incubation of Ross 308 chicken eggs. Eggs (n = 755) were incubated for 21 d. Quality analysis of fresh eggs was performed. Eggshells, albumen, and yolk were collected from fresh eggs and incubation d 1, 7, and 14. Eggshell thickness and strength, pH, vitelline membrane strength, fatty acid (FA) in the yolk, pH, viscosity, lysozyme activity, and crude protein content in thick albumen and amniotic fluid were analyzed. Hatching parameters were calculated. Egg weight loss was constant (8.04% overall). Lower egg surface temperature was found on d 7 compared to d 4, 14, and 18. A lower thickness of posthatch eggshells was found. The strength of the vitelline membrane significantly decreased within 24 h (by over 58%). During incubation, there was a decrease in thick albumen/amniotic fluid pH; an opposite trend was found in yolk pH. The vitelline membrane strength was negatively correlated with the albumen pH. Lysozyme activity was higher in fresh thick albumen and up to 2 wk of incubation. On d 7, the lowest activity was found in the amniotic fluid. On d 14, lysozyme activity increased in amniotic fluid. The higher viscosity of the thick albumen was demonstrated on d 7 and 14 of incubation. The lowest viscosity in amniotic fluid was found on the same days. Crude protein content was higher in thick albumen (d 7 and 14) and lowest in amniotic fluid on d 7. The FA content changed between d 0 and 14. The results indicate different use of FA, where PUFA decreased. Eggshell is used in the last week of incubation. The thick albumen is reduced, while the biological value of amniotic fluid is increasing. Lysozyme activity, viscosity, and crude protein content may be interdependent. It may indicate the flow of substances and the transfer of functions from the thick albumen to the amniotic fluid during chicken embryogenesis.
Topics: Animals; Chickens; Egg Shell; Muramidase; Amniotic Fluid; Ovum; Albumins; Fatty Acids; Embryonic Development; Egg Yolk; Eggs
PubMed: 37832191
DOI: 10.1016/j.psj.2023.103119 -
Journal of Morphology Sep 2023Ovarian follicles of sterlets (Acipenser ruthenus) are composed of a single oocyte surrounded by follicular cells (FCs), basal lamina, and thecal cells. Previtellogenic...
Asymmetry in previtellogenic and early vitellogenic oocytes, ultrastructure of follicular cells and egg envelope in the pigmented sterlet, Acipenser ruthenus L. 1758 (Chondrostei, Acipenseriformes).
Ovarian follicles of sterlets (Acipenser ruthenus) are composed of a single oocyte surrounded by follicular cells (FCs), basal lamina, and thecal cells. Previtellogenic oocytes are polarized. Homogeneous ooplasm (contains ribosomes) and granular ooplasm (comprises nuage aggregations of nuclear origin, rough endoplasmic reticulum (RER), Golgi complexes, ribosomes, and mitochondria) are distinguished. Granular ooplasm is initially located near the nucleus, contacts the plasma membrane of the oocyte (oolemma) and forms a thin layer underneath its entire perimeter. Next, a ring that surrounds the nucleus is formed and sends strands directed toward the oolemma. The lipid body composed of lipid droplets forms adjacent to this ring. Later, the granular ooplasm and strands enlarge toward the oolemma, lipid body disperses, and homogeneous ooplasm is no longer present. A thin cortical ooplasm is formed underneath the oolemma and does not contain any organelles. The oocyte nucleus moves to the center. The nucleoplasm contains lampbrush chromosomes, nuclear bodies, and multiple nucleoli. Early vitellogenic oocytes are polarized, too. Three regions in the ooplasm are distinguished: the perinuclear (contains lipid droplets near the nuclear envelope), the endoplasm (contains yolk platelets and lipid droplets), and the periplasm (contains yolk spheres, pigment granules, and microtubules). In all these regions the RER, Golgi complexes, nuage, and mitochondria are present. Micropinocytotic vesicles, Golgi vesicles and precursors of the internal layer of the egg envelope are in the cortical ooplasm. Some FCs delaminate from the follicular epithelium, degenerate and vesicles are released into the perioocytic space. They may contain precursors of egg envelope and may be involved in "cell-cell" communication. The egg envelope (zona radiata, zona pellucida) is made up of three layers: the vitelline envelope (inner layer), the middle layer, and the outer layer. In its deposition, both the oocyte and FCs are engaged.
Topics: Female; Animals; Oocytes; Ovarian Follicle; Fishes; Cytoplasm; Vitellogenesis
PubMed: 37585228
DOI: 10.1002/jmor.21631 -
Parasites & Vectors Aug 2023Stephanofilaria stilesi is a vector-borne filarioid nematode of cattle in North America that is transmitted via the hematophagous horn fly (Haematobia irritans)...
BACKGROUND
Stephanofilaria stilesi is a vector-borne filarioid nematode of cattle in North America that is transmitted via the hematophagous horn fly (Haematobia irritans) intermediate host. Despite being relatively common, little attention has been given to a thorough description of S. stilesi lesions and the potential integration of pathological and molecular diagnostic findings to confirm infection.
METHODS
To characterize the cutaneous lesions caused by S. stilesi in cattle (Bos taurus taurus and Bos taurus indicus), skin of the ventral abdominal midline was collected from 22 animals during postmortem examination. Skin samples were processed for histology, transmission electron microscopy (TEM), DNA extraction, PCR, and Sanger sequencing targeting molecular markers cytochrome oxidase c subunit 1 (cox1), 12S, 18S rDNA, and 28S rDNA.
RESULTS
Macroscopically, lesions ranged from 5 × 4 cm to 36 × 10 cm, consisting of one large single lesion, or two to four ovoid areas at the ventral abdominal midline, surrounding the umbilicus. Each lesion presented as ulcerative dermatitis with dry, serocellular crusts, or alopecic and lichenified areas. Histologically, eosinophilic, neutrophilic, and ulcerative dermatitis with furunculosis, folliculitis, and epidermal hyperplasia was observed. Cross sections of adult nematodes were identified in ~ 60% of the cases (n = 13) within intact follicles, sebaceous ducts, crusts, and areas of furunculosis. Stephanofilaria first-stage larvae (L1) were observed in five cases within "vitelline membranes" in the superficial dermis and crusts. Ultrastructurally, the L1 cross sections were compounded of smooth multilayered cuticle and somatic cells. The "vitelline membrane" is a tri-layered membrane where L1 are suspended in a matrix. Stephanofilaria stilesi DNA was found in 5 out of the 13 cases in which adults or L1 were histologically observed (38%) and in 1 out of the 9 cases without adults or L1 present (11%). Phylogenetic analyses suggest a closer relationship of the genus Stephanofilaria with Thelazioidea, instead of the family Filariidae (Filarioidea), in which it has been historically allocated.
CONCLUSIONS
Our study improved the characterization of lesions and described ultrastructural findings of S. stilesi and highlights that molecular tools should be utilized in combination with histology for improved diagnostic resolution.
Topics: Animals; Cattle; Phylogeny; Furunculosis; Filarioidea; Dermatitis; Muscidae; DNA, Ribosomal
PubMed: 37573424
DOI: 10.1186/s13071-023-05905-y -
Child's Nervous System : ChNS :... Jan 2024We previously developed a novel functional benchtop apparatus to simulate catheter occlusion in vitro utilizing avian vitelline membrane and chalaza to test catheter...
PURPOSE
We previously developed a novel functional benchtop apparatus to simulate catheter occlusion in vitro utilizing avian vitelline membrane and chalaza to test catheter designs and de-obstruction techniques. Here, we study the integration of double-lumen catheter-mediated backflow in the shunt system assembly and its potential for an in-line de-obstruction of an obstructed ventricular catheter.
METHODS
A double-lumen catheter was connected to a standard proximal shunt catheter for all trials. One limb of the double-lumen catheter was connected to the backflow mechanisms and allowed to loop back for fluid access. A micropump and a bi-corporal electromagnetic pump were utilized to provide various degrees of backflow at predetermined intervals. Flow rates were measured after initial occlusion and after implementation of the backflow mechanisms, and degrees of catheter blockage was calculated as a percentage of the unoccluded flow rate. Flow visualization was also used.
RESULTS
In baseline blockage of less than 50%, the average occluding agent weighed 0.3-0.6 g with baseline flow rates of 8.5-11.9 mL/min. After 5 min of backflow using a micropump, the degree of blockage was reduced in 50% of trials. Additional backflow for 5 min did not provide further improvements in flow rate. In baseline blockage of greater than 50%, the average occluding agent weighed 0.8-1.3 g with baseline flow rates of 1.1-4.2 mL/min. After 5 min of backflow, the system demonstrated a decreased blockage in 20% of trials; additional backflow for 5 min further improved the flow rate in 40% of the total trials. Only magnetic plates provided enough force to provide pulsatile backflow in the bi-corporal electromagnetic system.
CONCLUSIONS
The preliminary results of connecting a standard proximal catheter in series with a double-lumen catheter show a slight change in the percent occlusion from the baseline status several times when the retrograde flow occurred via one limb of the catheter. Additionally, the de-obstruction seems related to the length of the interval of the backflow and the initial percentage occlusion of the proximal catheter. The statistical analysis does not reveal a statistically significant reduction in occlusion in the proximal catheter with either backflow interval.
Topics: Humans; Catheters; Cerebrospinal Fluid Shunts; Prostheses and Implants; Hydrocephalus
PubMed: 37515721
DOI: 10.1007/s00381-023-06101-0 -
Pest Management Science Nov 2023Current mosquito-borne disease vector control strategies, largely based on chemical insecticides, are seriously threatened by increasing resistance worldwide. There is...
BACKGROUND
Current mosquito-borne disease vector control strategies, largely based on chemical insecticides, are seriously threatened by increasing resistance worldwide. There is also growing concerned about the adverse effects of insecticides on nontarget organisms and the environment, therefore effective and ecologically friendly alternative approaches are urgently needed. Targeting critical steps of reproduction is considered a potential way to control mosquito populations. Herein, we focused on the roles of chitin synthase A (encoded by chsa) in the reproduction of female mosquitoes.
RESULTS
The injection of small interfering RNA targeting Cpchsa in female Culex pipiens pallens (Diptera: Culicidae) had antireproductive effects, including decreased follicle numbers, egg-laying, and hatching rate. Scanning electron microscopy observations showed that Cpchsa silencing caused a defective egg envelope, including absence of the vitelline membrane and cracked chorion layers, which resulted in abnormal permeability. Widely distributed nurse cell apoptosis and follicular epithelial cell autophagy were observed in Cpchsa-silenced ovaries during the vitellogenesis phase. Consistent with the detective egg envelope formation during oogenesis, the exochorionic eggshell structures were also affected in eggs deposited by Cpchsa-silenced mosquitoes.
CONCLUSION
This study provided fundamental evidence for the role of chitin synthase A in the female reproductive process of mosquitoes and might result in a novel alternative strategy for mosquito control. © 2023 Society of Chemical Industry.
Topics: Animals; Female; Insecticides; Culex; Chitin Synthase; Mosquito Vectors; Reproduction; Culicidae
PubMed: 37409377
DOI: 10.1002/ps.7648 -
Nature Communications Jun 2023Embryonic tissues undergoing shape change draw mechanical input from extraembryonic substrates. In avian eggs, the early blastoderm disk is under the tension of the...
Embryonic tissues undergoing shape change draw mechanical input from extraembryonic substrates. In avian eggs, the early blastoderm disk is under the tension of the vitelline membrane (VM). Here we report that the chicken VM characteristically downregulates tension and stiffness to facilitate stage-specific embryo morphogenesis. Experimental relaxation of the VM early in development impairs blastoderm expansion, while maintaining VM tension in later stages resists the convergence of the posterior body causing stalled elongation, failure of neural tube closure, and axis rupture. Biochemical and structural analysis shows that VM weakening is associated with the reduction of outer-layer glycoprotein fibers, which is caused by an increasing albumen pH due to CO release from the egg. Our results identify a previously unrecognized potential cause of body axis defects through mis-regulation of extraembryonic tissue tension.
Topics: Animals; Down-Regulation; Chickens; Blastoderm; Embryonic Development
PubMed: 37277340
DOI: 10.1038/s41467-023-38988-3 -
Bioengineering (Basel, Switzerland) May 2023The lipid layer surrounding the vitelline membrane of insect eggs has a critical role in the waterproofing and desiccation resistance of embryos. However, this lipid...
The lipid layer surrounding the vitelline membrane of insect eggs has a critical role in the waterproofing and desiccation resistance of embryos. However, this lipid layer also prevents the flux of chemicals into the embryos, such as cryoprotectants, which are required for successful cryopreservation. The permeabilization studies of silkworm embryos remain insufficient. Therefore, in this study, we developed a permeabilization method to remove the lipid layer in the silkworm, , and examined factors affecting the viability of dechorionated embryos, including the types and exposure times of chemicals and embryonic stages. Among the chemicals used, hexane and heptane were effective for permeabilization, whereas Triton X-100 and Tween-80 were less effective. Regarding the embryonic stages, there were significant differences between 160 and 166 h after egg laying (AEL) at 25 °C. Consequently, we found that the treatment of 160 AEL embryos with hexane for 30 s was the best condition for the permeability and viability of embryos, in which over 62% of the permeabilized embryos grew up to the second larval instar and their moths could lay fertilized eggs. Our method can be used for various purposes, including permeability investigations using other chemicals and embryonic cryopreservation.
PubMed: 37237633
DOI: 10.3390/bioengineering10050563