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Studying avian embryology necessitates reliable and precise staging tables-descriptions of embryonic features appearing during development that are used to approximate the extent of embryonic development from fertilisation to hatching. Staging tables for waterfowl (Anseriformes) have previously been established based on morphological features from fertilisation to approximately 10 days before hatching. Embryonic changes over the final 10 days of pre-hatching development have also been documented and proposed as useful staging criteria. However, the reliability of these changes-which focus on the size of the bill and middle toes-as useful staging criteria across different waterfowl breeds has not been fully examined. To evaluate the reliability of these criteria for staging near-hatching embryos, we examined 27 embryos of Mallard (Anas platyrhynchos) and Swan Goose (Anser cygnoides). Comparisons with previously published data revealed that size variation within the same developmental stage across breeds is equivalent to within-breed variation across different stages, suggesting limited reliability of bill and middle toe size for staging waterfowl embryos. Consequently, we devised novel staging criteria for waterfowl based on four easily measurable morphological traits and show that these criteria allow correct stage identification with over 70% accuracy. Our results highlight the importance of quantifying staging accuracy for improving the reliability of embryonic staging tables., (© 2025 The Author(s). Journal of Morphology published by Wiley Periodicals LLC.)

The goose industry represents a significant sector within the broader waterfowl industry in China, with early gonadal development playing a pivotal role in enhancing population productivity. Melatonin plays a crucial role in regulating early gonadal development, but the molecular mechanism underlying the effect of melatonin as a regulator remains unclear. In this study, the relationship between melatonin and egg production performance in Jilin white goose was first established, then, an in Ovo injection of melatonin in goose embryos on the embryonic day 12 (E12) was performed to evaluate the impact of melatonin on the gonadal development in Jilin white goose. The results showed that the in Ovo injection of melatonin increased the expression of FOXL2 in the gonads while decreasing the expression of CYP19A1. Transcriptome analyses revealed that a total of 1184 differentially expressed genes were screened in Jilin white goose embryonic gonads among the two comparison groups. These DEGs were enriched in Gene Ontology (GO) terms related to the microtubule cytoskeleton, cytoskeleton, cell adhesion, and biological adhesion and the KEGG enrichment analysis suggested that the DEGs were mainly enriched in "MAPK signaling pathway", "Wnt signaling pathway", "Calcium signaling pathway", "Focal adhesion", "Neuroactive ligand-receptor interaction", "Melanogenesis", "mTOR signaling pathway", "ECM-receptor interaction", and "TGF-beta signaling pathway" in the two comparison groups. The protein-protein interaction network analysis (PPI) indicated that selected DEGs, such as PDIA3, ATCB, PKM, PGAM1, NME2, LDHB, and CALM2, were highly related to the regulation of the reproduction system development. Additionally, the expression trends of 4 identified DEGs were validated by RT-qPCR. In conclusion, this study elucidates the potential regulatory role of melatonin in gonadal development in geese, identifying candidate genes involved in early gonadal differentiation and maturation. Furthermore, the findings offer valuable practical implications for the goose industry, providing professionals with scientifically grounded strategies to enhance reproductive efficiency and overall population productivity., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing Interests: Yongfeng Sun reports financial support was provided by Jilin Provincial Science and Technology Department, Jilin Animal Husbandry Administration and Jilin Provincial Department of Human Resources and Social Security., (Copyright © 2024. Published by Elsevier Inc.)

The present investigation was aimed at predicting a still (i.e., dead) vs. live embryo within a hatching goose egg by measuring the eggshell cooling rate. For this, we daily measured the temperature (T) values on the shell surface of goose eggs after they were removed from the incubator and during further natural cooling. T was recorded every 0.5 h for further 1.5 h of cooling. It was possible to recognize eggs with dead embryos using the combination of T, egg weight (W), and surface area (S). The resultant indicator (TS/W) was called specific temperature index (STI). The mathematical relationship using STI measurements between Days 8-13 facilitated 80 % correct identification of the eggs with dead embryos. Additionally, we derived mathematical dependencies for shell weight (W s ) and thickness (t) by utilizing the values of W, egg volume (V), S, the average T of all measurements taken, as well as the drop in T during 1.5 h of natural cooling. The key advantage of these parameters was their measurement and/or calculation by applying non-destructive methods. The integrated application of these parameters resulted in achieving high calculation accuracy as judged by correlation coefficients of 0.908 for W s and 0.593 for t. These novel mathematical models have the potential to decrease hatching waste by predicting embryo viability. Our research will add to a toolkit for non-invasive egg assessment that is useful in the poultry industry, research on eggs, and engineering., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Low hatchability has been a persistent challenge in the goose industry. Establishing standard atlases and comprehending embryonic development patterns are essential to improving the hatching rates of goose eggs. However, comprehensive descriptions of normal atlases, embryonic development, and energy requirements in geese are lacking. In this study, a total of 120 fertile eggs from well-known large Shitou goose were incubated using 12 nesting purebred female geese. During hatching, both the temperature of the eggshells and the weight of eggs were recorded, and daily photographs of the embryos were captured to monitor their development closely. After hatching, counted the number of pores per unit area of eggshells by choosing eggs from without sperm, dead embryos, and normally hatched. Furthermore, 150 Shitou goose eggs were hatched by automatic incubator, with adjustments made based on observed normal developmental stages that incubated by female geese. The eggs were carefully opened to meticulously document embryonic morphology and create a detailed development map. Measurements were taken of the eye diameter, length of the lower beak, tarsometatarsus bone, and embryo length. Subsequently, an analysis was conducted to assess the calcium, phosphorus, crude protein, and crude fat content to study the energy requirements for embryo development. characteristics on the 7th, 15th, 23rd and 28th days of Shitou goose hatching corresponded to the 5th, 10th, 17th and 19th days of chicken egg incubation, respectively. These days were distinguished individually by "visible embryo's eye", "closure", "sealing the door", and "flashing hair". Besides, the hatch rate of the incubator reached 86.67%, and the cumulative water loss rate increased with embryo age. Notably, normally developing embryos displayed a significantly higher number of pores on the eggshell surface compared to dead embryos (P < 0.05). Additionally, embryonic body length, eyeball diameter, and lower beak length exhibited continuous growth until day 19 of incubation, while tarsometatarsus length increased steadily from days 12 to 31. Liver size measurement began on the 10th day of incubation, while both leg and chest muscles showed continuous growth from the 12th day. For energy demand, the embryo primarily relied on protein sourced from the egg yolk within the first 10 days of development. Afterward, the egg yolk provided both protein and fat for embryonic growth. In summary, this study has generated a comprehensive developmental map for Shitou goose embryos, offering valuable insights into their growth and morphological changes throughout the incubation period. This map can serve as a reference for optimizing machine incubation techniques to enhance goose egg hatching rates and provide fresh perspectives on the development of geese., Competing Interests: DISCLOSURES The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

1. Melanin distribution typically exhibits a gradient dilution along the dorsal-ventral axis of the body, including in domestic geese. However, the specific genes and molecular mechanisms responsible for this melanin distribution pattern remain incompletely understood.2. The transcriptomic comparisons were conducted at three embryonic stages, specifically on embryonic d 15 (E15), 22 (E22), and 29 (E29), between the pigmented dorsal skin and the depigmented distal foot.3. Differentially expressed genes (DEGs) associated with melanin synthesis were identified, particularly TYR , TYRP1 , and EDNRB2 , which exhibited significantly higher expression levels in the dorsal skin at E15 and E22. However, expression levels significantly decreased in later stages (E29).4. The ASIP gene showed remarkably high-expression levels in the distal feet compared to the dorsal skin post-E22 stage (log 2 FC: 5.31/6.88 at E22/E29). Gene Ontology (GO) enrichment analysis detected eight terms associated with melanin synthesis and melanosome formation ( p  < 0.05), including melanosome membrane (GO: 0033162) and melanin biosynthetic process (GO: 0042438). Additionally, KEGG pathway analysis showed significant enrichment of the melanogenesis pathway (hsa004916) at d 22 (E22).

1. The Wulong goose is a Chinese breed and a source of high-quality meat and eggs. A characteristic of the Wulong goose is that a proportion of the birds do not have eyelids, known as the Huoyon trait.2. Wulong geese exhibiting the Huoyan trait at embryonic stages of 9 days (E9), 12 days (E12) and 14 days (E14) were selected alongside those with normal eyelids for comprehensive transcriptome sequencing. Differentially expressed gene (DEG) and functional enrichment analyses were performed and finally, eight DEG were chosen to verify the accuracy of qPCR sequencing.3. Overall, 466, 962 and 550 DEG were obtained from the three control groups, D9 vs . N9, D12 vs . N12 and D14 vs . N14, respectively, by differential analysis ( p  < 0.05). CDKN1C , CRH , CROCC and TYSND1 were significantly expressed in the three groups. Enrichment analysis revealed the enrichment of CROCC and TYSND1 in pathways of cell cycle process, endocytosis, microtubule-based process, microtubule organising centre organisation, protein processing and protein maturation. CDKN1C and CRH were enriched in the cell cycle and cAMP signalling pathway.4. Some collagen family genes were detected among the DEGs, including COL3A1 , COL4A5 , COL4A2 and COL4A1 . FREM1 and FREM2 genes were detected in both Huoyan and normal eyelids. There was a significant difference ( p  < 0.01) in FREM1 expression between ED9 and ED14 in female embryos, but this difference was not observed in male embryos.

In this study, the effects of Goose parvovirus (GPV) infection as well as the possible role of NS1 protein on apoptosis induction in goose embryo fibroblast (GEF) cells were examined. Flow cytometry analysis and TUNEL assays revealed that GPV infection and NS1 transfection induced significant apoptosis in GEF cells compared to what was observed in mock-infected cells. Interestingly, the increase in the rate of apoptosis detected in GPV-infected GEFs was accompanied by an increased viral load in the cells. In addition, the apoptotic pathway was mediated by apoptosis-inducing factors (AIFs) and internal factors that influence the release of AIFs. The results indicated that the mitochondrial membrane potential was decreased, and AIF expression was increased in the nucleus (P < 0.01). Reactive oxygen species (ROS) increased gradually within 48 h (P < 0.001). Cathepsin D activities were also increased (P < 0.05). The results demonstrated that the AIF-mediated pathway is a new mitochondrial apoptotic pathway and that mitochondrial depolarization, ROS content, and cathepsin D activities are the key factors influencing apoptosis in GEF cells., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The primary feather follicles are universal skin appendages widely distributed in the skin of feathered birds. The morphogenesis and development of the primary feather follicles in goose skin remain largely unknown. Here, the induction of primary feather follicles in goose embryonic skin (pre-induction vs induction) was investigated by de novo transcriptome analyses to reveal 409 differentially expressed genes (DEGs). The DEGs were characterized to potentially regulate the de novo formation of feather follicle primordia consisting of placode (4 genes) and dermal condensate (12 genes), and the thickening of epidermis (5 genes) and dermal fibroblasts (17 genes), respectively. Further analyses enriched DEGs into GO terms represented as cell adhesion and KEGG pathways including Wnt and Hedgehog signaling pathways that are highly correlated with cell communication and molecular regulation. Six selected Wnt pathway genes were detected by qPCR with up-regulation in goose skin during the induction of primary feather follicles. The localization of WNT16, SFRP1 and FRZB by in situ hybridization showed weak expression in the primary feather primordia, whereas FZD1, LEF1 and DKK1 were expressed initially in the inter-follicular skin and feather follicle primordia, then mainly restricted in the feather primordia. The spatial-temporal expression patterns indicate that Wnt pathway genes DKK1, FZD1 and LEF1 are the important regulators functioned in the induction of primary feather follicle in goose skin. The dynamic molecular changes and specific gene expression patterns revealed in this report provide the general knowledge of primary feather follicle and skin development in waterfowl, and contribute to further understand the diversity of hair and feather development beyond the mouse and chicken models., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In order to enrich the Anser cygnoides genome and identify the gene expression profiles of primary and secondary feather follicles development, de novo transcriptome assembly of skin tissues was established by analyzing three developmental stages at embryonic day 14, 18, and 28 (E14, E18, E28). Sequencing output generated 436,730,608 clean reads from nine libraries and de novo assembled into 56,301 unigenes. There were 2,298, 9,423 and 12,559 unigenes showing differential expression in three stages respectively. Furthermore, differentially expressed genes (DEGs) were functionally classified according to genes ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and series-cluster analysis. Relevant specific GO terms such as epithelium development, regulation of keratinocyte proliferation, morphogenesis of an epithelium were identified. In all, 15,144 DEGs were clustered into eight profiles with distinct expression patterns and 2,424 DEGs were assigned to 198 KEGG pathways. Skin development related pathways (mitogen-activated protein kinase signaling pathway, extra-cellular matrix -receptor interaction, Wingless-type signaling pathway) and genes (delta like canonical Notch ligand 1, fibroblast growth factor 2, Snail family transcriptional repressor 2, bone morphogenetic protein 6, polo like kinase 1) were identified, and eight DEGs were selected to verify the reliability of transcriptome results by real-time quantitative PCR. The findings of this study will provide the key insights into the complicated molecular mechanism and breeding techniques underlying the developmental characteristics of skin and feather follicles in Anser cygnoides ., (Copyright © 2020 Liu et al.)

To determine whether geese are susceptible to infection by avian leukosis virus (ALV), 702 serum samples from domestic and foreign goose breeds were screened for p27 antigen as well as being inoculated into DF-1 cell cultures to isolate ALV. Although 5.7% of samples were positive for p27 antigen, reactivity appeared to be non-specific because no ALV was detected in the corresponding DF-1 cultures. To further determine whether geese are susceptible to ALV-J isolated from chickens, ALV-J strain JS09GY7 was artificially inoculated into 10-day-old goose embryos, with one-day-old hatched goslings then screened for p27 antigen and the presence of ALV. In all cases, the results of both tests were negative. Liver tissues from the 1-day-old goslings were screened using a polymerase chain reaction-based assay, which failed to amplify ALV-J gene fragments from any of the samples. Further, no histopathological damage was observed in the liver tissues. ALV-J was further inoculated intraperitoneally into one-day-old goslings, with cloacal swabs samples and plasma samples then collected every 5 days for 30 days. All samples were again negative for the presence of p27 antigen and ALV, and liver tissues from the challenged geese showed no histopathological damage and were negative for the presence of ALV-J gene fragments. Furthermore, p27 antigen detection, PCR-based screening, and indirect immunofluorescence assays were all negative following the infection of goose embryo fibroblasts with ALV-J. Together, these results confirm that virulent chicken-derived ALV-J strains cannot infect geese, and that p27 antigen detection in goose serum is susceptible to non-specific interference.

Embryo development and chick quality are influenced by parental genotype, age, nutrition, environment, and flock management. The aim of study was to determine if genotype, age of goose or eggs laid near the onset of egg production vs. eggs laid near the end of reproduction influence the stage of embryo at oviposition. Three experiments were undertaken. To compare genotypes (Experiment 1) 150 eggs were collected from 3-year-old commercial line White Koluda (WK) geese and from two breeds involved in a genetic resources conservation program, Zatorska (Za) and Bilgoraj (Bi). Age comparison (Experiment 2) was conducted with 200 eggs collected from 1-, 2-, 3-, and 4-year-old WK geese. To compare laying periods (Experiment 3), 150 WK eggs were collected at the first week of March and 100 at the second half of June. Eggs were stored for 72 h at 16°C, staged using Eyal-Giladi and Kochav (EGK, Roman numerals) and Hamburger and Hamilton (HH, Arabic numerals) procedures. Experiment 1: Individual breed differences were evident with Stage X EGK embryos comprising 42.4, 33.3, and 38.7% in the eggs examined from the WK, Bi, and Za, respectively. For all breeds combined, 38.8% of the embryos were in Stage X, but in the next order in WK there was stage XI (18.2%), while in geese from the genetic reserve it was stage XIII (Bi - 33.3; Za - 29.0%). Experiment 2: In eggs of 1-, 2-, and 3-year-old WK geese, the majority of embryos (38.7, 32.4 and 42.2%, respectively) were in Stage X. In contrast, in 4-year-old geese the embryos were in Stage XI (36.1%). Experiment 3: In eggs collected in March and in June most of embryos were in Stage X (33.7% and 43.6%, respectively). In addition, more developmentally advanced stages (XI-XIII) were similar in both periods. However, embryos in Stage 2 HH were only observed in eggs collected at the end of laying season. Interestingly, earlier stages (VI-IX) were observed exclusively in the eggs collected in March., (© 2019 Poultry Science Association Inc.)

The aim of this study was to characterize the embryonic ontogeny of 5-hydroxyindoles and 5-methoxyindoles synthesis pathways in the goose pineal organ. The study was performed on embryos aged 14-28 days, which have been incubated under a 12L:12D cycle. The pineal organs were collected for measurements of indole content by HPLC every 6 h on embryonic day (ED) 14, ED 16, ED 18 and ED 22 or every 2 h on ED 24, ED 26 and ED 28. The level of tryptophan showed no significant changes during development and no day-night variations. The content of 5-hydroxytryptophan increased between ED 14 and ED 26. It was significantly higher during scotophase than during photophase starting from ED 14. The serotonin content was low during the early stages of development (ED 14-ED 18) and prominently increased from ED 20. The serotonin levels also showed day-night differences; however, they were less conspicuous than those of 5-hydroxytryptophan. The changes in the level of 5-hydroxyindole acetic acid were similar to those of serotonin. 5-Hydroxytryptophol was measurable from ED 18. Levels of N -acetylserotonin, which were detectable for the first time on ED 16, prominently increased between ED 22 and ED 28 and showed significant day-night differences from ED 20. Melatonin was detectable from ED 18. Like N -acetylserotonin, its content increased rapidly between ED 22 and ED 28, and from ED 20 showed diurnal variations. 5-Methoxyindole acetic acid and 5-methoxytryptophol occurred at measurable levels from ED 18 and ED 26, respectively. The obtained results showed that embryonic development of indole metabolism in the goose pineal organ starts with the beginning of serotonin synthesis. The processes of serotonin acetylation and 5-hydroxyindoles methylation were turned on later. Diurnal rhythmicity develops very early in the embryonic pineal organ of the goose when the eggs are incubated under a 12 h light: 12 h dark schedule. Two processes are responsible for generation of the diurnal rhythms of 5-hydroxyindoles and 5-methoxyindoles: (i) hydroxylation of tryptophan and (ii) acetylation of serotonin.

Embryos from aquatic birds are the primary models for the study of flipper development. While some staging of early embryogenesis in duck have been studied, characterization of the stages of the entire embryonic development period in water birds has not been described. This study aimed to establish a comparison of complete morphological development staging for ducks (Anas platyrhynchos) and geese (Anser cygnoides) with the embryonic staging system by Hamburger and Hamilton (HH) for the chicken (Gallus gallus). Our results show that morphological development in the chicken, duck, and goose are similar in the early stages. The major differences occurred after stage 27 of embryonic development, where the beak shape in ducks and geese was wider and longer than in chickens. In addition, the second and third interdigital webs of the hind limb of the chicken were found to be degraded from stage 31, and eventually vanished at stage 35; however, they were retained in ducks and geese. Rapid physical development occurred in the mid-to-late stages in ducks and geese. To our best knowledge, this is the first description of complete embryonic development for the duck and goose. Establishment of an embryonic staging system for duck and goose provides new models for the study of waterfowl development., (© 2019 Poultry Science Association Inc.)

The phenomenon of multiple embryos in birds and other oviparous vertebrates is very rare and thus poorly studied. Some populations of wild birds or lines of poultry, including the zatorska goose, may be particularly predisposed to developing monovular twin embryos (MTE). The aim of the study was to investigate the frequency of single yolk twin embryos in the zatorska goose and to establish whether these embryos are generated as a result of polyembryony or other developmental anomalies.Embryopathological analysis, which included identification of the developmental stages of all MTEs at the time of death, was carried out on 1,955 unhatched eggs obtained from 20 pedigree breeding flocks of the zatorska goose. Post-mortem examinations revealed as many as 13 MTEs (0.66% compared to embryonated eggs) at varying developmental stages.The results of genotyping of the microsatellite loci of 7 pairs of twins demonstrated genetic similarity in 6 of them; this suggests that they developed from 1 blastoderm, while differences in 2 loci of 1 pair show that the MTE originates from 2 blastodiscs. Pedigree analysis of the MTEs does not provide evidence that predisposition toward twinning results from the genetic background. However, more frequent MTE occurrence could be seen in eggs stored for more than 8 d. It seems that embryos of the zatorska goose are more susceptible to duplication under special conditions, such as prolonged egg storage.

The objective of this study was to evaluate the changes in the goose embryo transcriptome during feather development. RNA-Sequencing (RNA-Seq) was used to find the transcriptome profiles of feather follicles from three stages of embryonic dorsal skin at embryonic day 13, 18, and 28 (E13, E18, E28). The results showed that 3001, 6634, and 13,780 genes were differently expressed in three stages. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that differentially expressed genes (DEGs) in E13 vs. E18 were significantly mapped into the GO term of extracellular structure organization and the pathway of extracellular matrix (ECM)-receptor interaction. In E18 vs. E28, the top significantly mapped into GO term was the single-organism developmental process; the pathway was also the ECM-receptor interaction. DEGs in E13 vs. E28 were significantly mapped into the GO term of the multicellular organismal process and the pathway of cell adhesion molecules. Subsequently, the union of DEGs was categorized by succession cluster into eight profiles, which were then grouped into four ideal profiles. Lastly, the seven genes spatio-temporal expression pattern was confirmed by real-time PCR. Our findings advocate that interleukin 20 receptor subunit alpha ( IL20RA ), interleukin 6 receptor ( IL6R ), interleukin 1 receptor type 1 ( IL-1R1 ), Wnt family member 3A ( WNT3A ), insulin-like growth factor binding protein 3 ( IGFBP3 ), bone morphogenetic protein 7 ( BMP7 ), and secreted-frizzled related protein 2 ( SFRP2 ) might possibly play vital roles in skin and feather follicle development and growth processes.

The aim of this study was to estimate the genetic parameters and inbreeding effect on the fertility, embryo mortality and hatchability traits in the Zatorska goose covered by the animal genetic resources conservation program. The material for this study contains information about results of hatching of 18 863 eggs from 721 dams and 168 sires, laid between 1998-2015. Genetic parameters were estimated based on the threshold animal model by the use of Restricted Maximum Likelihood and Gibbs sampling. The percentage of fertilized eggs ranged yearly between 37-80%. The percentage of embryo mortality was very low, ranging between 4.63-23.73%. The percentage of the hatched goslings from the total number of analyzed eggs was on average 33.18%, and 53.72% from fertilized eggs. On average based on both methods, the heritability estimates of the fertility, embryo mortality and hatchability reached 0.36, 0.07, 0.24 for males and 0.44, 0.11, 0.32 for females. The genetic trend had increasing tendency for fertility and hatchability and was stable for embryo mortality for both sexes. The obtained result shows that the Zatorska goose can be still maintained in the reserves of the local gene pool according to current rules and use in the local market as a breed with good reproductive potential., (© 2018 Japanese Society of Animal Science.)

Normal tables provide an objective step-wise description of the morphological development of an embryo. Such tables have been described for the chicken, turkey, quail, and duck embryos, but there is no such staging table for goose embryos. As the goose has one of the longest incubation periods of all the poultry species and embryo mortality during incubation is relatively high, a normal table of goose embryo development would be useful in assessing the morpho-genetic status of the goose embryo before and during incubation. In this study, embryos were isolated from commercial White Koluda goose eggs stored no longer than four days in a cool room (18°C) prior to incubation and after 4, 8, 12, and 16 h of incubation. Embryo staging was based on the normal tables described for the chicken by Eyal-Giladi and Kochav (EGK) and Hamburger and Hamilton (HH). Goose embryos from unincubated eggs were at Stage X and XI EGK and after 16 h of incubation the majority of embryos were between Stages 2 and 4 HH. Our results suggest that while the stage of development of the embryo in the unincubated goose egg is similar to that reported for the chicken, although the diameter of goose embryo is slighter larger. Following incubation, a goose embryo advances more slowly than a chicken embryo up to 16 h of incubation., (© 2016 Poultry Science Association Inc.)

Three types of mechanical papillae, i.e., conical, filiform, and hair-like papillae, are present on the tongue in the domestic goose. Within conical papillae, we distinguish three categories: large and small conical papillae on the body and conical papillae on the lingual prominence. The arrangement of mechanical papillae on the tongue in Anseriformes is connected functionally with different feeding mechanisms such as grazing and filter-feeding. The present work aims to determine whether morphology of three types of mechanical papillae in goose at the time of hatching is the same as in an adult bird and if the tongue is prepared to fulfill feeding function. Our results revealed that the primordia of the large conical papillae start to develop during the differentiation stage. The primordia of the small conical papillae and conical papillae of the lingual papillae start to develop during the growth stage. At the end of the growth stage, only large conical papillae, three pairs of small conical papillae, and conical papillae of the lingual prominence have similar arrangement as in an adult bird. The shape and arrangement of the remaining small conical papillae probably will be changed after hatching. During embryonic period, the filiform papillae and hair-like papillae are not formed. The embryonic epithelium that covered the mechanical papillae undergoes transformation leading to the formation of multilayered epithelium. During prehatching stage, epithelium becomes orthokeratinized epithelium. In conclusion, the tongue of the domestic goose after hatching is well prepared only for grazing. The filtration of food from water is limited due to the lack of filiform papillae.

Background: Muscovy duck parvovirus (MDPV) is the etiological agent of Muscovy duckling parvoviral disease, which is characterized by diarrhea, locomotive dysfunction, stunting, and death in young ducklings, and causes substantial economic losses in the Muscovy duck industry worldwide. FZ91-30 is an attenuated vaccine strain that is safe and immunogenic to ducklings, but the genomic information and molecular mechanism underlining the attenuation are not understood., Methods: The FZ91-30 strain was propagated in 11-day-old embryonated goose eggs, and viral particles were purified from the pooled allantoic fluid by differential centrifugation and ultracentrifugation. Single-stranded genomic DNA was extracted and annealed to form double-stranded DNA. The dsDNA digested with NcoI resulted two sub-genomic fragments, which were then cloned into the modified plasmid pBluescript II SK, respectively, generating plasmid pBSKNL and pBSKNR. The sub-genomic plasmid clones were sequenced and further combined to construct the plasmid pFZ that contained the entire genome of strain FZ91-30. The complete genome sequences of strain FM and YY and partial genome sequences of other strains were retrieved from GenBank for sequence comparison. The plasmid pFZ containing the entire genome of FZ91-30 was transfected in 11-day-old embryonated goose eggs via the chorioallantoic membranes route to rescue infectious virus. A genetic marker was introduced into the rescued virus to discriminate from its parental virus., Results: The genome of FZ91-30 consists of 5,131 nucleotides and has 98.9 % similarity to the FM strain. The inverted terminal repeats (ITR) are 456 nucleotides in length, 14 nucleotides longer than that of Goose parvovirus (GPV). The exterior 415 nucleotides of the ITR form a hairpin structure, and the interior 41 nucleotides constitute the D sequence, a reverse complement of the D' sequence at the 3' ITR. Amino acid sequence alignment of the VP1 proteins between FZ91-30 and five pathogenic MDPV strains revealed that FZ91-30 had five mutations; two in the unique region of the VP1 protein (VP1u) and three in VP3. Sequence alignment of the Rep1 proteins revealed two amino acid alterations for FZ91-30, both of which were conserved for two pathogenic strains YY and P. Transfection of the plasmid pFZ in 11-day-old embryonated goose eggs resulted in generation of infectious virus with similar biological properties as compared with the parental strain., Conclusions: The amino acid mutations identified in the VP1 and Rep1 protein may contribute to the attenuation of FZ91-30 in Muscovy ducklings. Plasmid transfection in embryonated goose eggs was suitable for rescue of infectious MDPV.

Avian hepadnaviruses are etiological agents of hepatitis B, that has been identified primarily in ducks, and more recently in various avian species. In this paper, 16 hepadnaviruses were detected by polymerase chain reaction (PCR) in the field samples from dead embryos of commercially reared domestic duck and goose. Based on the molecular analysis of the S-protein gene sequences and phylogenetic Neighbor-joining tree, identified viruses were clustered in the same genetic group, indicating no host-related diversity. Both duck and goose-origin hepadnaviruses were grouped within the cluster consisting of "Western-country" and "Chinese" duck hepatitis B (DHBV) isolates, showing more evolutionary distances with other known avian hepadnaviruses. Histopathologically, the lesions observed in the liver tissue from hepadnavirus positive duck and goose embryos varied from low to mild degree of perivascular mononuclear cells and mixed cell infiltrations, followed by mild vacuolar changes. Small focal necrotic changes in the liver parenchyma, and bile ductular proliferation were also found in examined liver samples. Generally, the microscopic findings resemble those described in experimentally infected ducks, while this was the first description of hepadnavirus associated lesions in domestic goose. Although hepadnaviruses are considered to have a very narrow host range, this study showed that domestic ducks and geese are susceptible to infection with genetically almost identical hepadnaviruses, that were likely to produce similar microscopic changes in the liver of both duck and goose embryos. The impact of naturally occurred hepadnavirus infection and possible synergistic interactions with other infectious or non-infectious agents on embryo viability needs further investigation., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Adrenergic control of cardiovascular function is a common component of regulation in embryonic birds studied to date. Our goal was to investigate adrenergic and cholinergic cardiovascular regulation in two goose species, the Canada goose (Branta canadensis) and the related domestic goose (Anser anser domesticus) to determine if these species possess similar regulation during embryonic development. We determined mean arterial pressure (Pm) and heart rate (fH) responses to serial blockade of cholinergic, β-adrenergic and α-adrenergic receptors in B. canadensis and A. domesticus at 70 and 90% of total incubation (28days). Both species possessed cholinergic and β-adrenergic tone on fH at 70% of incubation that increased in intensity with development. In addition, a constant α-adrenergic tone on Pm was present in both species. Our findings indicate that these geese species possess both cholinergic and adrenergic cardiovascular regulation providing information needed for speculation regarding the commonalities in the ontogeny of these cardiovascular regulatory elements in birds., (© 2013.)

The general development of the tongue in birds was described by Lillie (1908) in chicken. Bryk et al. (1992) also studied the tongue development in chicken and they observed development of the conical papillae of the body. Our study aims to describe the timing of the development of the tongue morphological features in the domestic goose by using SEM methods. The tongue of the domestic goose is characterized by the widest variety of shape of the particular part of the tongue and mechanical papillae. Results indicated that the formation of the apex, body, lingual prominence, and the root of the tongue take place between the 10th and 19th day of incubation. The tongue elongates rapidly between the 16th and 18th day of incubation. Simultaneously, the median groove appears on the body and the lingual prominence and elongates towards the rostral part of the tongue. The conical papillae of the tongue develop gradually. On the body, the conical papillae develop from the caudal part of the body to the rostral part and on the lingual prominence from the median part of the prominence to the lateral part. Hair-like papillae at the caudal surface of the body of the tongue remain primordial to the end of the incubation. Our studies on the morphogenesis of the tongue in the domestic goose revealed changes in shape of the particular part of the tongue and rapid pace of the formation of mechanical papillae. The tongue is completely develop before hatching and ready to collect food., (Copyright © 2012 Wiley Periodicals, Inc.)

In birds, downy feather quantity mainly affected by the follicles. Wnt6, a secreted cysteine-rich protein, plays a key role in follicular development as an intercellular signaling molecule. The present study was to investigate the follicle development and Wnt6 polymorphism in Wanxi-white geese, a Chinese indigenous breed. In total, 300 fertilized eggs were hatched. At embryonic stage and on early birth goslings, the diameter and density of follicles from different sites were examined after sectioning and staining. The results showed that the diameter of primary feather follicles in thorax, venter, dorsum and flank had no difference at embryonic stage. In contrast, after birth, thorax and ventral feather follicles had greater diameter than those on dorsum and flank. Similarly, the primary feather follicle density was higher in thorax and venter than in dorsum and flank at embryonic stage. The secondary feather follicle diameter in flank was greater than that in other sites examined. The secondary follicle showed lush growth in E27 with thickest in ventral and thorax. Overall, follicle formed consistently in dorsal and flank, and follicle in thorax and ventral formed in another consistent way. The polymorphism study showed 2 single nucleotide polymorphisms of Wnt6 and 3 genotypes identified. Sequencing revealed two nucleotide transitions, T451C and A466G, which were synonymous mutations causing codons for aspartate and lysine to change from GAU to GAC and from AAA to AAG, respectively. This information about follicle development and Wnt6 polymorphisms would provide potential utilization in marker-assisted selection program for down feather selection.

The invariant chain (Ii) plays a critical role in MHC class II-associated chaperone representing antigen peptide and receptor. In this study, we cloned goose Ii cDNA by 5' and 3' rapid amplification of cDNA ends and tested its mRNA expression in the different tissues by fluorescent quantitative PCR. Two Ii isoforms, GIi-1and GIi-2, were 1,204 and 1,393 bp in length, respectively, and were found in all tested tissues. The expression of GIi in all tested tissues of 2-mo-old geese was higher than in that of embryos and in immune tissues was higher than in other tissues. The comparison of Ii sequences among pigeon, zebra finch, duck, chicken, quail, mouse, human, Pan troglodytes, and Salmo salar in the phylogenetic tree showed that they were classified into 5 lineages. Ii contains 5 functional domains. Further comparison between full-length Ii and each of the 5 domains of 6 species indicated a distinct difference. In 3-dimensional structures some amino acid residues located at critical sites in Ii structural region were identical or similar, which suggests that these residues were important in maintaining Ii essential secondary structures such as β-turn.

1. Evidence has accumulated in mammals to support the idea that mitochondrial DNA (mtDNA) deletions and mutations might contribute to ageing and reproductive failure. White Roman geese were monitored to evaluate the effect of large-scale deletions of mtDNA in an avian species. 2. A total of 340 samples from 114 dead embryos, 111 weak goslings, and 115 normal goslings were used in this experiment. The regions of these two large-scale mtDNA deletions, ΔmtDNA6829 and ΔmtDNA6992, were between the COI and ND5 genes. A 3·6% (4 out of 111) positive sample was detected in the weak goslings. In contrast, no large-scale mitochondrial DNA deletions were detected in either the dead embryos (0 out of 114) or the normal goslings (0 out of 115). 3. Large-scale mtDNA deletions may be a factor causing weak goslings.

Results concerning the contents of retinol in the liver, residual yoke of 25-day embryos and yoke of eggs depending on the level of vitamins A, D3 and E in the diet of geese by grey Obroshin breeds in reproductive period are presented in the paper. It is established, that vitamin D3 reduces the level of retinol deposition in the tissues of embryos and yoke of eggs of geese, and addition of vitamins A and E to a diet of geese raises the level of retinol both in the liver and residual yoke of embryos, and in yokes of geese eggs. Besides the data about changes of fatty-acid spectrum of common lipids and contents of lipid peroxidations products in tissues of the liver and pectoral muscles of 25-day embryos are presented in the paper depending on the level of vitamins A, D3 and E in geese diet during their reproductive period. Introduction of vitamin A--in quantity of 10000 IU, vitamin D3--in quantity of 3000 IU, in the composition of mixed fodder of geese during the reproductive period and vitamin E in quantity 35 IU on 1 kg to mixed fodder optimizes fatty-acid structure of the common lipids and the level of peroxidations lipids products in the liver and pectoral muscles of embryos.

beta-Catenin signaling has been reported to initiate feather bud development. In the present study, beta-catenin gene was isolated and identified from a cDNA library constructed using embryonic goose skin. Expression patterns of beta-catenin gene in the dorsal skin of goose embryos were investigated using the methods of semi-quantitative reverse transcription PCR, Northern blot analysis, and in situ hybridization. The sequence of beta-catenin was found highly conserved at the amino acid level, sharing 100, 99, and 99% identity with chicken, Chinese soft-shell turtle, and human sequences, respectively. Relatively high levels (62.51 +/- 7.11% to 101.74 +/- 7.29%) of beta-catenin mRNA were detected in the dorsal skin samples. The levels of beta-catenin expression were most prominent at the early stage from embryo day (E)10 to E20 and then significantly declined with the embryonic development. In situ hybridization demonstrated that at E10, beta-catenin expression was mainly observed at the surface periderm cells and the localized region of the epidermal layer. Because feather bud forms with an anterior-posterior orientation, strong staining was observed in the periderm layer and in the ectoderm and epidermis with a diffuse distribution within the internal area of the buds. The stronger staining was seen in the barb ridges than in the center pulp of the feather follicles at E18 and E20. In this study, expression of Shh as a marker gene for the bud development was examined paralleling with expression patterns of beta-catenin. It was found that the expression pattern of beta-catenin was almost similar spatially and temporally to that of Shh mRNA at the later stages of bud development. The differential beta-catenin mRNA expression in the goose dorsal skin may be essential for promoting the normal development of embryonic feather bud.

In the present study, the process of feather follicle formation in the Zi goose, a Chinese indigenous breed, was investigated during various stages of embryonic development by using a modified histological processing method. The results showed that the feather placodes evolved initially at embryonic day (E) 12 on the spinal feather tract, emerging as symmetrical structures. Sequentially, the buds elongated from E14 to E16 with anterior-posterior and proximal-distal asymmetries, and invaginated to form the primary feather follicles, which were identified to develop the contour feathers or remiges. The remarkable observation at this stage was the formation of the feather follicle wall, which was understood to be the result of the epidermis surrounding the base and further invaginating into the dermis. With the differentiation of the barbule plates, the various types of feathers were determined. We proved that the secondary feather follicles simply had radially symmetrical barb ridges, with much smaller diameters than the primary follicles, and that they developed only downy feathers. The primary and secondary follicles evolved independently of each other and formed ranks in a linear fashion. Moreover, quantitative measurements of the densities of both follicles confirmed that the density of the primary follicles sharply reached the maximum at E18, and then decreased gradually. Coincidentally, the secondary follicles started to increase from the age of E18, and up to E26 the density of the secondary follicles exceeded that of the primary follicles. Each of the primary feather follicles was richly encircled with muscles, which pointed to a quadrangularly arranged network in the dermis. The present work lays the foundation for further study of the cellular and molecular mechanisms of feather follicle morphogenesis in geese.

Goose embryos were infected with goose haemorrhagic polyomavirus (GHPV) onto the chorioallantoic membrane (CAM) in order to examine the effect of GHPV on the embryos and to obtain data on whether embryos could develop into infected, virus-shedding goslings, as well as to present an accurate biological method for virus titration. The reported method of infection could offer a possibility to express the virus titre as the median embryo infective dose (EID(50)). As a special pathological feature of the disease, extensive cerebral haemorrhages were observed, which protruded the skullcap in many cases. Some embryos infected with 10(1.25) or 10(0.25) EID(50)/0.2 ml were able to hatch; however, they were in poor physical condition and died by post-hatching day 4 showing haemorrhagic nephritis and enteritis of geese. Virus shedding was revealed by polymerase chain reaction. The ability of some of the infected goose embryos to hatch may indicate the potency of GHPV to spread vertically, although this needs further study for confirmation.

Peculiarities of antioxidant homeostasis of geese brain tissue during embryogenesis and early postnatal period have been studied. It has been shown that the cerebrum and hindbrain tissues are characterized by a higher level of lipid peroxidation compared to liver. Main antioxidative enzymes' activity (superoxide dismutase, catalase, glutathione peroxidase) in the brain already reaches its maximum in the middle period of embryogenesis. We have found that brain tissues are characterized by a lower activity of intracellular enzymes (superoxide dismutase, catalase) but increased glutathione peroxidase activity as compared to liver. The rate of Fe2+ initialized lipid peroxidation and coefficient of antioxidative activity were used as a criterion for evaluation of antioxidative system's status. According to the dynamics of these factors the highest tension of antioxidative system in the brain appears in the period of the contour (28 days) and juvenile (49 days) feather formation.

Dynamics of fatty acid composition of geese liver lipids in embryogenesis and early postnatal period has been investigated. High nonsaturation of these tissues lipids is shown during all the period of researches. It is established, that in the second half of embryogenesis and in the beginning of postnatal period (first two weeks) the increase of oleic acid contents is observed with simultaneous decrease of the contents of the polynonsaturated fatty acids (PUFA) showing properties of endogenous bioregulators (arachidonic acid (AA), docosahexaenoic acid [figure: see text] (DHA). From the two-weeks age of gosling the contents of the mentioned acids changes in the opposite direction. The coordination of dynamics of TBA-active products with nonsaturation of fatty acids (FA) expressed in mEqu/g concerning double bonds, and also with respect to AA/DHA has been revealed.

The development of the cecum and colon in the goose was investigated during the period from the 15th to 28th day of the incubation and from 1 to 30 days of age after hatching by light microscopy. By day 15 of the incubation, in the cecum and colon, the lumen was surrounded by pseudostratified epithelium. The previllous ridges appeared at 15th and 17th days of the incubation in the colon and ceca, respectively. At the base of previllous ridges, the epithelium changed into a simple prismathic epithelium at 15th and 17th days of the incubation in the colon and cecum, respectively. The villi appeared at the 21st days of the incubation. The crypts and goblet cells appeared on the first day after hatching. In the pre-hatching period, the lamina muscularis mucosa was present only in the colon. The submucosa consisted of loosely aggregated connective tissue in the pre-hatching period. In the post-hatching period, it consisted of a very thin layer of connective tissue. Its presence was only obvious where the cells of the submucosal nerve plexus or occasional large blood vessels considerably increased its thickness. The nerve plexus corresponding to the Auerbach's plexus of the mammalian intestine and submucosal nerve plexus appeared by 15th days of the incubation. From the 15th to 28th day of incubation, the tunica muscularis consisted of circular smooth muscle cells in the ceca. On the 28th day of the incubation a thinner longitudinal muscle layer added to the circular muscle layer. In the colon there was an outer longitudinal and a thicker circular muscle layer.

The features of antioxidant protection of tissues of a liver and blood of the gooses in embriogenesis and early postnatal ontogenesis are found out. Maximal contents TBA active products both in a liver, and in a blood are observed in 28 diurnal embriones. Is shown, that in a liver the activity of basic antioxidant enzymes (superoxide dismutases, catalase and glutathione peroxidase) in a liver is developed already at early stages embriogenesis and is considerably enlarged in the end embriogenesis. The becoming of enzymatic system of a blood descends much more slower.

Hatchability of 69,324 fertilized eggs of White Italian geese from the WD1 maternal strain, selected for egg production, and of 25,226 eggs from the WD3 paternal strain, selected for weight gain, were compared. A significant difference was noted, but its magnitude varied during the reproduction season. Mean results for the WD1 strain (80.9%) were always higher (P < 0.01) than those of WD3 strain (75.8%). To determine the causes of this difference, goose eggs from WD1, WD3, and the Kuban (K) strain that demonstrate high reproductive performance (egg number, fertility, and hatchability) were incubated in a laboratory scale incubator. Embryonic mortality and egg weight loss during incubation were determined. Best hatchability (78.5%) was noted in eggs from K goose strain and the weight loss attained 10.9% of the initial egg weight, until Day 25 of incubation. With the higher egg weight loss in Strains WD1 (11.8%) and WD3 (13.2%), hatchability was diminished to 67.3 and 65.5%, respectively. Better hatchability resulted from lower mortality up to Day 6 of incubation and, especially, between Days 7 and 25 of incubation. The differences found in the egg weight losses and in embryonic mortality suggest a need for adjusting the incubation technique to the particular goose genotype. Embryo cultivation in vitro in plastic containers made the comparison of embryo survival possible, because the interaction of incubator microclimate by eggshell quality was eliminated. The results noted due to strain were ranked in the following order: K, WD1, and WD3. The experimental findings suggest that, apart from that of shell quality effect on various egg weight losses during incubation, there appear to be other factors that cause differences in hatchability of the goose strains studied.

1. The purpose of this work was to test the possibility of increasing the hatchability of goose eggs with low mass specific eggshell gas conductance (Gsp), by drilling holes through the eggshell into the air cell, and thus solving both the low water loss rate and low oxygen availability problems. 2. A linear relationship was found between the area of a hole drilled and the apparent increase in eggshell gas conductance (G). Drilling more than one hole increased apparent G 3-6 times more than one hole only, of the same total area. 3. Hole-drilling did not increase egg contamination. The drilling of a 5 mm2 hole on day 17 of incubation increased hatchability both in laboratory tests and in commercial hatcheries (6.1% and 10.5% respectively). 4. Drilling holes on days 15 to 22 of incubation increased hatchability when the predicted mean water loss was lower than 14%. Drilling on day 25 did not have a significant effect, and drilling on day 11 of incubation was too early. 5. Drilling a hole into the aircell (during the second half of incubation) may increase hatchability of low conductance eggs, although oxygen pressure under the eggshell should then be checked in order to evaluate oxygen availability to the embryo.

A total of 240 embryonated goose eggs obtained from two susceptible flocks were used. Half of the eggs were inoculated into the allantoic cavity with a virulent strain (7593) of duck plague virus isolated from an acute outbreak, and the other half were inoculated with the attenuated vaccine virus (KAPEVAC). Ten, 100 or 1000 CPU/0.1 ml virus were given on days 12 and 20 of incubation. Embryos that died and surviving embryos killed at 5-day intervals were examined by light and electron microscopy. The yolk and the serum of embryos that survived until hatching were assayed for antibody content. Lymphocytes separated from the blood were used for the immuno-rosette formation and lymphocyte stimulation tests. Pathomorphological changes indicative of virus replication occurred in the liver, kidney, myocardium, gizzard muscle and chorioallantoic membrane (CAM) of the embryos in the case of both virus strains. The time of onset and severity of these changes and the time and rate of embryonic mortality depended on the virulence of the strain used for inoculation, the virus dose and the time of inoculation. Virus-neutralizing (VN) antibodies were demonstrable neither in the yolk nor in the serum of goslings exsanguinated after hatching. The lymphocytes recognized the virus antigen in the in vitro cellular tests and responded to it with blastogenic transformation. As opposed to adult birds, in the embryos duck plague virus infection did not cause damage to the digestive tract mucosa and the lymphoid organs.

The present work is concerned with the study of myosin fractions prepared from the hatching muscle (m. complexus) and a control muscle (m. pectoralis) of the developing goose embryo. The m. complexus attained its maximum mass at hatching and in the 4-day-old bird the mass of this muscle was only one fourth of that recorded at hatching. The m. complexus was hypertrophied already on the 21st day. At days 21, 27 and 28 of incubation and at posthatching days myosin preparations were made from both muscles. Partial purification of myosins from both sources yielded a high molecular weight fraction characteristic of the adult bird and one other protein fraction with molecular mass half of myosin. Both preparations exhibited the characteristic properties of myosin. The lower molecular weight fraction was also shown to develop filamentous aggregates as did the higher molecular-weight, gel filtrated myosin. The phosphate content of the half molecular mass myosin fraction prepared from the embryonic m. complexus at days prior to hatching was considerably higher than that of the high molecular weight fraction and the predominant component was P-Arg. Since the embryonic myosin was still not available in the m. complexus of the 4-day-old birds and the hypertrophied muscle underwent regression after hatching it appears that this myosin fraction is actively involved in breaking through the shell during the hatching period in geese.

Cells from a goose embryo were shown to release particle-associated RNA-directed DNA polymerase and RNase H activities that required the presence of Nonidet P-40 for detection. The particles were not infectious and did not have endogenous DNA synthesis. The goose particle DNA polymerase was related to the DNA polymerase of spleen necrosis virus with respect to size and was inhibited by immunoglobulin G to spleen necrosis virus DNA polymerase. However, goose cells producing DNA polymerase-containing particles did not contain reticuloendotheliosis virus-related nucleotide sequences in their DNA.

A light microscopic study of serial sections of the hearts of 64 avian embryos (chicken, turkey, duck and goose) was undertaken in order to elucidate the development of the atrioventricular bundle and its branches. The developing atrioventricular bundle and its branches were shown to appear simultaneously (at about a third of the way through incubation) as relatively large, discrete fasciculi, which were dark-staining when compared with developing cardiac muscle. Prior to the appearance of the bundle and branches, the developing conducting tissue was found to be present as a widespread precursor tissue, which was distributed as a complete, dark-staining subendocardial sleeve, lining the whole of the ventricular canal, ventriculobulbar loop, and extending into the common atrial cavity. From these observations, it is concluded that, with septation and chamber formation, the tissue of the sleeve is partly incorporated within the chamber walls to give origin to diffuse Purkinje tissue, and is partly retained as the bundle and branches. The manner of derivation of the bundle and branches is discussed, and it is stressed that the definitive positions of the bundle and branches denote the junctional zones of the components of septation.