Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (14): 2837-2849.doi: 10.3864/j.issn.0578-1752.2022.14.013

• ANIMAL SCIENCE·VETERINARY SCIENCE • Previous Articles     Next Articles

Dynamic Changes of Yolk Microbiota in Yellow-Feathered Broiler and Its Role on Early Colonization of Intestinal Microbiota During the Embryonic Stage

DING Peng1(),TONG YueYue1,LIU HuiChao1,YIN Xin1,LIU JiangJun2,HE Xi1,SONG ZeHe1,ZHANG HaiHan1()   

  1. 1. College of Animal Science and Technology, Hunan Engineering Research Center of Poultry Production Safety, Engineering Research Center for Feed Safety and Efficient Utilization of Ministry of Education, Hunan Co-Innovation Center for Livestock and Poultry safety, Hunan Agricultural University, Changsha 410128
    2. YongZhou Vocational Technical College, Yongzhou 425000, Hunan
  • Received:2021-06-16 Accepted:2022-05-01 Online:2022-07-16 Published:2022-07-26
  • Contact: HaiHan ZHANG E-mail:shkdingpeng@163.com;zhhous@163.com

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Abstract:

【Objective】 The purpose of this experiment was to study the dynamic changes and functions of the yolk microbiota during the hatching of yellow-feathered broiler fertilized eggs, and the influence of yolk microbiota on the early intestinal colonization of embryos. 【Method】 One hundred fertilized eggs of 35-week-old yellow-feathered broilers were selected and placed in automatic incubators for incubation. Before hatching, the incubator was cleaned and disinfected, and the internal ventilation system and humidity control system were checked for normal operation. The incubation temperature was controlled at (37.5±1) ℃ and the humidity at (60±10)%. Six breeder eggs were randomly selected on days 7, 11, 15 and 19 of incubation to collect the yolk samples by stripped the eggshell and allantoic membrane in a sterile operation table, exposed the embryo to the yolk sac and then pierced it through a sterile syringe. Samples were rapidly snap frozen in liquid nitrogen after collection and subsequently transferred to a -80 ℃ refrigerator for storage pended test. The yolk microorganisms were subjected to DNA extraction, and 16s rDNA sequencing and sequence analysis were performed. 【Result】 1) A total of 4 305 OTUs were annotated in the yolk of fertilized eggs, and a total of 18 Phyla, 34 Classes, 53 Orders, 89 Families and 126 Genera were found in the yolk and intestine at different incubation time points. The phylum level mainly included Proteobactreia, Firmicutes, Bacteroidetes, Verrucomicrobia, etc. The genus level is dominated by Pelomonas, Ralstonia, Aquabacterium, Faecalibacterium, etc; 2) Significant differences in the Chao1 index (P = 0.027) and Shannan index (P = 0.043) were observed in the Alpha diversity of yolk microbiota at different incubating stages. The yolk microbiota tended to stabilize with incubation time, while the relative abundance of Akkermansia in the yolk increased continuously to 0.53%, 0.84%, 1.43% and 3.10% at E07, E11, E15 and E19, respectively. At the genus level, the yolk microbiota mainly included Pelomonas, Ralstonia, Aquabacterium and so on; 3) The functions of yolk microbiota in yellow-feathered broiler breeding eggs mainly include carbohydrates metabolism, amino acids metabolism, lipids metabolism, cofactors and vitamins metabolism; 4) Both the yolk microbiota and intestinal microbiota of fertilized eggs were dominated by Proteobacteria, Firmicutes, Bacteroidetes, and Verrucomicrobia at the phylum level, and Pelomonas, Ralstonia, Aquabacterium and Akkermansia at the genus level. 【Conclusion】There are abundant microbial populations in the yolk of yellow-feathered broiler fertilized eggs, which play an important role in the absorption and metabolism of yolk nutrients and the early microbial colonization of embryo intestine.

Key words: yellow-feather broiler, embryo, yolk, intestine, nutritional metabolism

Fig. 1

Composition of yolk microbiota and intestinal microbiota in the chicken embryo and the abundant microbes at each level (phylum, class, order, family, and genus)"

Fig. 2

Embryonic yolk microbial difference at different developmental stages of yellow-feathered broilers A: PCA analysis: Each point represents a sample, and points of different colors indicate different groups; B: alpha diversity analysis: Each panel corresponds to an alpha diversity index, which is identified in the top area. In each panel, the abscissa is the group label, and the ordinate is the value of the corresponding alpha diversity index; C: The relative abundance of yolk microbiota at the phylum level; D: The relative abundance of yolk microbiota at the genus level; E: Venn: The Venn diagram shows the core microbes shared at different stages of chicken embryo development; F: LEfSe analysis: Taxonomic cladogram generated from LEfSe showing landmark microbiota profile of 4 stages of chicken embryo development. CL represents yellow-feathered broiler, Y represents yolk, and E represents embryonic stages"

Fig. 3

Functional profiles of the microbial community in yolk of yellow-feathered broilers. The abscissa is the abundance of the functional pathway, the ordinate is the functional classification of KEGG in the second level, and the rightmost is the first level of the pathway"

Fig. 4

Comparison of embryonic microbiota between yolk and intestine of yellow-feathered broilers A: PCA analysis: Each point represents a sample, and points of different colors indicate different groups; B: alpha diversity analysis: Each panel corresponds to an alpha diversity index, which is identified in the top area. In each panel, the abscissa is the group label, and the ordinate is the value of the corresponding alpha diversity index; C: The relative abundance of yolk microbiota and intestine microbiota at the phylum level; D: The relative abundance of yolk microbiota and intestine microbiota at the genus level; E: LEfSe analysis: Taxonomic cladogram generated from LEfSe showing landmark microbiota profile of 4 stages of chicken embryo development. CL represents yellow-feathered broiler, Y represents yolk, and E represents embryonic stages, I represents intestine"

Fig. 5

Early changes of intestinal microbiota in chicken embryo of yellow-feathered broilers A: PCA analysis: Each point represents a sample, and points of different colors indicate different groups; B: alpha diversity analysis: Each panel corresponds to an alpha diversity index, which is identified in the top area. In each panel, the abscissa is the group label, and the ordinate is the value of the corresponding alpha diversity index; C: The relative abundance of intestine microbiota at the phylum level; D: The relative abundance of intestine microbiota at the genus level; E: LEfSe analysis: Taxonomic cladogram generated from LEfSe showing landmark microbiota profile of 4 stages of chicken embryo development. CL represents yellow-feathered broiler, and E represents embryonic stages, I represents intestine"

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