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Veterinary Microbiology Dec 2020Since 2017, duck spleen necrosis caused by new variant duck orthoreovirus (N-DRV) infection had been observed in several provinces in China. This disease retards the...
Since 2017, duck spleen necrosis caused by new variant duck orthoreovirus (N-DRV) infection had been observed in several provinces in China. This disease retards the growth and development of ducks, thereby reducing feed return rate. N-DRV infection causes damage to duck spleen and other immune organs, leading to immunosuppression and susceptibility to other pathogens. In this study, we successfully constructed a breeding duck artificial infection model and found that N-DRV infection can cause pathologic changes, such as ovarian hemorrhage, follicle atrophy, and fallopian tube bleeding in breeding ducks, resulting in significantly reduced fertilization rate and egg hatching rate. Viral RNA was present in egg vitelline membrane, duck embryo, and duckling's spleen samples, as determined through quantitative polymerase chain reaction (qPCR). Autopsy revealed obvious pathologic changes in the spleen and other organs, although there were no obvious early clinical symptoms observed in ducklings. Sequence distance and phylogenetic analysis confirmed that N-DRV-SD19 re-isolated from the spleen samples of ducklings was consistent with the strain N-DRV-XT18 used for infecting breeding ducks. The findings in this study confirmed that N-DRV can be vertically transmitted through eggs, which provide an important reference for the disease prevention and control.
Topics: Animals; Ducks; Female; Infectious Disease Transmission, Vertical; Male; Orthoreovirus, Avian; Ovum; Phylogeny; Poultry Diseases; RNA, Viral; Reoviridae Infections; Sequence Analysis, DNA; Spleen
PubMed: 33059276
DOI: 10.1016/j.vetmic.2020.108861 -
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 -
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 -
Cells Apr 2022In sea urchin, the immediate contact of the acrosome-reacted sperm with the egg surface triggers a series of structural and ionic changes in the egg cortex. Within one...
In sea urchin, the immediate contact of the acrosome-reacted sperm with the egg surface triggers a series of structural and ionic changes in the egg cortex. Within one minute after sperm fuses with the egg plasma membrane, the cell membrane potential changes with the concurrent increases in intracellular Ca levels. The consequent exocytosis of the cortical granules induces separation of the vitelline layer from the egg plasma membrane. While these cortical changes are presumed to prevent the fusion of additional sperm, the subsequent late phase (between 1 and 4 min after fertilization) is characterized by reorganization of the egg cortex and microvilli (elongation) and by the metabolic shift to activate de novo protein and DNA syntheses. The latter biosynthetic events are crucial for embryonic development. Previous studies suggested that the early phase of fertilization was not a prerequisite for these changes in the second phase since the increase in the intracellular pH induced by the exposure of unfertilized sea urchin eggs to ammonia seawater could start metabolic egg activation in the absence of the cortical granule exocytosis. In the present study, we have demonstrated that the incubation of unfertilized eggs in ammonia seawater induced considerable elongations of microvilli (containing actin filaments) as a consequence of the intracellular pH increase, which increased the egg's receptivity to sperm and made the eggs polyspermic at fertilization despite the elevation of the fertilization envelope (FE). These eggs also displayed compromised Ca signals at fertilization, as the amplitude of the cortical flash was significantly reduced and the elevated intracellular Ca level declined much faster. These results have also highlighted the importance of the increased internal pH in regulating Ca signaling and the microvillar actin cytoskeleton during the late phase of the fertilization process.
Topics: Actin Cytoskeleton; Ammonia; Animals; Hydrogen-Ion Concentration; Male; Sea Urchins; Zygote
PubMed: 35563801
DOI: 10.3390/cells11091496 -
Poultry Science Sep 2020A study was conducted to determine differences between Histomonas meleagridis-infected and control pullets based on disease signs, hen growth, and egg production and...
A study was conducted to determine differences between Histomonas meleagridis-infected and control pullets based on disease signs, hen growth, and egg production and quality. Ross 708SF females were weighed and then placed in pens on the day of hatch (92 chicks/pen). At 25 D, 4 pens were infected with H. meleagridis in the cloaca, whereas 4 pens were control. At 5, 10, and 20 D after inoculation, 5 birds per pen (2 birds per pen at 20 D) were subjectively scored for blackhead disease. Birds were feed restricted based on BW and/or egg production. Individual BW were collected at 3, 5, 13, 15, 20, and 64 wk. Egg production was recorded at 24-63 wk. Egg quality was measured at 30, 34, 39, 42, and 56 wk and included shell and vitelline membrane strength, shell thickness, egg weight, and Haugh units. Hatchability was measured at 27, 37, and 60 wk and fertility at 27 and 37 wk. Treatment effects were determined by JMP Pro 14 using GLM with means separated using the Student t test (P ≤ 0.05). Cecal lesions were apparent on 5, 10, and 20 D and liver lesions on 10 and 20 D for the infected birds. The control had no histomoniasis lesions. Flock uniformity differed on wk 13 and 20 (P = 0.04; 0.04). Infected birds weighed less at 64 wk (P = 0.002). The onset of lay was not delayed. Infected birds produced more eggs during 1 period (P = 0.02). The infected birds produced heavier eggs at 30 wk (P = 0.04), eggs with a stronger and thicker shell at 42 wk (P = 0.05, 0.03), and eggs with a stronger vitelline membrane at 56 wk (P = 0.049). Hatchability and fertility did not differ (P > 0.05). H. meleagridis was observed in the infected birds' cecal samples at trial termination. This study indicates early infection with H. meleagridis has limited effects on pullet egg production and quality.
Topics: Animals; Body Weight; Chickens; Female; Fertility; Oviposition; Poultry Diseases; Protozoan Infections, Animal; Trichomonadida
PubMed: 32867968
DOI: 10.1016/j.psj.2020.05.020 -
Insect Biochemistry and Molecular... Mar 2024Chitinases (CHT) comprise a large gene family in insects and have been classified into at least eleven subgroups. Many studies involving RNA interference (RNAi) have...
Functional importance of groups I and II chitinases, CHT5 and CHT10, in turnover of chitinous cuticle during embryo hatching and post-embryonic molting in the red flour beetle, Tribolium castaneum.
Chitinases (CHT) comprise a large gene family in insects and have been classified into at least eleven subgroups. Many studies involving RNA interference (RNAi) have demonstrated that depletion of group I (CHT5s) and group II (CHT10s) CHT transcripts causes lethal molting arrest in several insect species including the red flour beetle, Tribolium castaneum, presumably due to failure of degradation of chitin in their old cuticle. In this study we investigated the functions of CHT5 and CHT10 in turnover of chitinous cuticle in T. castaneum during embryonic and post-embryonic molting stages. RNAi and transmission electron microscopic (TEM) analyses indicate that CHT10 is required for cuticular chitin degradation at each molting period analyzed, while CHT5 is essential for pupal-adult molting only. We further analyzed the functions of these genes during embryogenesis in T. castaneum. Real-time qPCR analysis revealed that peak expression of CHT10 occurred prior to that of CHT5 during embryonic development as has been observed at post-embryonic molting periods in several other insect species. With immunogold-labeling TEM analysis using a fluorescein isothiocyanate-conjugated chitin-binding domain protein (FITC-CBD) probe, chitin was detected in the serosal cuticle but not in any other regions of the eggshell including the chorion and vitelline membrane layers. Injection of double-stranded RNA (dsRNA) for CHT5 (dsCHT5), CHT10 (dsCHT10) or their co-injection (dsCHT5/10) into mature adult females had no effect on their fecundity and the resulting embryos developed normally inside the egg. There were no obvious differences in the morphology of the outer chorion, inner chorion and vitelline membrane among eggs from these dsRNA-treated females. However, unlike dsCHT5 eggs, dsCHT10 and dsCHT5/10 eggs exhibited failure of turnover of the serosal cuticle in which the horizontal chitinous laminae remained intact, resulting in lethal embryo hatching defects. These results indicate that group I CHT5 is essential for pupal-adult molting, whereas group II CHT10 plays an essential role in cuticular chitin degradation in T. castaneum during both embryonic hatching and all of the post-embryonic molts. CHT10 can serve in place of CHT5 in chitin degradation, except during the pupal-adult molt when both enzymes are indispensable to complete eclosion.
Topics: Female; Animals; Tribolium; Coleoptera; Chitinases; Chitin; Molting; Insect Proteins
PubMed: 38295884
DOI: 10.1016/j.ibmb.2024.104087 -
Journal of Agricultural and Food... Sep 2020To explore the thermally induced alterations in chicken egg vitelline membrane (CEVM) protein abundances, a comparative proteomic analysis of CEVM after 10 days of...
To explore the thermally induced alterations in chicken egg vitelline membrane (CEVM) protein abundances, a comparative proteomic analysis of CEVM after 10 days of storage at 30 °C was performed. Altogether, 981 proteins were identified, of which 124 protein abundances were decreased and 79 were increased. Bioinformatic analysis suggested that the altered proteins were related to structure ( = 10), mechanical properties ( = 13), chaperone ( = 15), antibacterial ( = 12), and antioxidant ( = 3). Alterations in abundances of structural proteins, possibly resulting from the disintegration of these complexes, were observed in this study, suggesting a loss in fibrous structure. Several proteins involved in mechanical strength ( = 10), elasticity ( = 3), and chaperone were decreased in abundances, which indicated that deficits in these proteins might affect the CEVM mechanical properties. These findings will extend our understanding of CEVM deterioration during high-temperature storage from a proteomic perspective.
Topics: Animals; Chickens; Egg Proteins; Eggs; Food Storage; Hot Temperature; Proteomics; Vitelline Membrane
PubMed: 32809818
DOI: 10.1021/acs.jafc.0c03538 -
International Journal of Biological... Dec 2020The chicken egg vitelline membrane (CEVM) is an important structure for the transmembrane transport of egg yolk components, protection of the blastodisc, and separation...
The chicken egg vitelline membrane (CEVM) is an important structure for the transmembrane transport of egg yolk components, protection of the blastodisc, and separation of egg white and egg yolk. In this study, the N-glycoproteome of the CEVM was mapped and analyzed in depth. Total protein of the CEVM was digested, and the glycopeptides were enriched by a hydrophilic interaction liquid chromatography microcolumn and identified by nano liquid chromatography/tandem mass spectrometry. A total of 435 N-glycosylation sites on 208 N-glycoproteins were identified in CEVM. Gene Ontology enrichment analysis showed that CEVM N-glycoproteins are mainly involved in the regulation of proteinases/inhibitors and transmembrane transport of lipids. Mucin-5B is the primary N-glycoprotein in the CEVM. Comparison of the main N-glycoproteins between the CEVM and other egg parts revealed the tissue specificity of N-glycosylation of egg proteins. The results provide insights into protein N-glycosylation in the chicken egg, CEVM functions and underlying mechanisms.
Topics: Animals; Chickens; Chromatography, Liquid; Egg Proteins; Gene Ontology; Glycoproteins; Mucin-5B; Tandem Mass Spectrometry; Vitelline Membrane
PubMed: 32860793
DOI: 10.1016/j.ijbiomac.2020.08.193 -
Veterinary Microbiology May 2020In 2019, a novel goose astrovirus (GoAstV) epidemiological investigation on geese was conducted in Shandong province, China. During the investigation, a high prevalence...
In 2019, a novel goose astrovirus (GoAstV) epidemiological investigation on geese was conducted in Shandong province, China. During the investigation, a high prevalence of novel GoAstV was observed in symptom-free breeding geese flocks. Moreover, the novel GoAstV-specific RNA was detected in either breeder birds or their progenies. To verify the hypothesis that the novel GoAstV could be transmitted vertically, a total of 42 WuLong breeder geese, aged 335 days, were equally divided into three groups for experimental infection. The SDPY isolate of novel GoAstV (A/goose/Shandong/SDPY/2018, SDPY), preserved in our laboratory, was injected intramuscularly to subjects of group A while orally inoculated to those of group B. After the inoculation, novel GoAstV RNA was detected in vitelline membrane, embryos, and allantoic fluid of goose embryos in novel GoAstV infected groups. Moreover, the ORF2 gene of novel GoAstV from vitelline membrane, embryo, allantoic fluid as well as gosling shared almost 100 % nucleotide homology to a novel GoAstV virus isolated from the goose ovary which produced the egg, suggesting that the novel GoAstV can be vertically transmitted in the goose. Taken together, the findings provide evidence of possible vertical transmission of novel GoAstV from breeding goose to goslings.
Topics: Animals; Astroviridae; Astroviridae Infections; China; Female; Geese; Infectious Disease Transmission, Vertical; Male; Phylogeny; Poultry Diseases; RNA, Viral
PubMed: 32402337
DOI: 10.1016/j.vetmic.2020.108657 -
Theriogenology Jul 2020The objective of this review is to provide new insights into the possible use of a proteomic method known as Intact Cell Matrix-Assisted Laser Desorption-ionization... (Review)
Review
The objective of this review is to provide new insights into the possible use of a proteomic method known as Intact Cell Matrix-Assisted Laser Desorption-ionization Time-Of-Flight Mass Spectrometry (ICM-MS) in animal clinical research. Here, we give an overview of the basics of this technique, its advantages and disadvantages compared with other proteomic approaches, past applications and future perspectives. A special emphasis on its implementation in animal reproduction science is given, including examples of the reliable use of ICM-MS on fertility screening. In mammals, the ICM-MS profiles from pig epididymal spermatozoa reflect the proteome changes that they undergo during epididymal maturation and could be associated with the acquisition of fertilizing ability. In chicken, using adequate pre-processing and bioinformatics analysis tools, sperm ICM-MS profiles showed characteristic spectral features that allowed their classification according to their actual fertilizing ability. The association of ICM-MS and Top-down proteomic strategies allowed the identification of chicken fertility biomarkers candidates such as protein vitelline membrane outer layer protein 1 (VMO-1) and avian beta-defensin 10 (AvBD10). In female reproduction, a similar approach on ovarian follicular cells allowed the identification of specific markers of oocyte maturation in the oocyte and surrounding cumulus cells. Altogether, these results indicate that ICM-MS profiling could be a suitable approach for molecular phenotyping of male and female gametes.
Topics: Animals; Gene Expression Regulation; Livestock; Proteomics; Reproduction; Single-Cell Analysis; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 32284210
DOI: 10.1016/j.theriogenology.2020.02.037