Changes in the Diversity and Composition of Gut Microbiota of Red-Crowned Cranes (Grus japonensis) after Avian Influenza Vaccine and Anthelmintic Treatment

Abstract

Gut microbiota homeostasis is important for host health and well-being; however, drugs may affect the composition and function of the gut microbiota. Red-crowned cranes are a vulnerable species. Treatment of red-crowned cranes with avian influenza vaccines and anthelmintics has played pivotal roles in therapeutic management in zoos. To investigate the changes in the diversity and composition of gut microbiota after the avian influenza vaccine and anthelmintic treatment, we used 16S rRNA sequencing to obtain and compare the bacterial community composition before and after the treatment. The alpha diversity of the gut microbiota of red-crowned cranes decreased on the day of the treatment and then fluctuated over time. The composition of gut microbiota tended to be similar in the short term after the treatment, as supported by the beta diversity hierarchical cluster analysis. Only 3, 8, and 72 operational taxonomic units (OTUs) of the three individuals were shared among the five groups before and after treatment. The relative abundance of Firmicutes significantly increased to 99.04% ± 0.28% on the day of the treatment, in which the relative abundance of Lactobacillus was 93.33% ± 5.85%. KEGG pathways analysis indicated that the main function of the gut microbiota is involved in metabolism, and the present study indicates that the gut microbiota of red-crowned cranes is resilient to the avian influenza vaccine and anthelmintic, even disordered in the short term, and could recover over time. More individual experimentation and functional potential in metabolism are needed in the future to support animal disease control and optimal management in the zoo.

Keywords: anthelmintic; avian influenza vaccine; diversity and composition; gut microbiota; red-crowned crane.

Figure 1

Alpha diversity of the intestinal bacterial community of the samples of red-crowned cranes. Comparison of the alpha diversity (Chao1 and Shannon) of the intestinal bacterial community of red-crowned cranes before and after the avian influenza vaccine and anthelmintic treatment.

Figure 2

Beta diversity hierarchical cluster analysis of microbial genera of the fecal samples in different groups (B, D0, D5, D10, D15) of red-crowned cranes. The panel on the left is a hierarchical clustering tree diagram, in which samples are clustered according to their similarity with each other. The panel on the right is the abundance ranking Stacked histogram of the top 10 genera.

Figure 3

Venn diagrams. Venn diagrams show the numbers of OTUs (97% sequence identity) that were shared or not shared of the fecal samples of each individual (S1, S2, and S3) and shared OTUs between different groups (B and D0; D0 and D5; D5 and D10; D10 and D15).

Figure 4

Differences in relative abundance of microbial phyla and genera of the fecal samples of red-crowned cranes in different groups (B, D0, D5, D10, D15). Differences in abundance of dominant microbial phyla (a: top 10 are shown) and genera (b: top 15 are shown) of five groups (B, D0, D5, D10, D15) of each individual (S1, S2, and S3).

Figure 5

Distributions of primary and secondary metabolic pathways of all fecal samples of red-crowned cranes annotated by KEGG pathways. Pie chart represents primary metabolic pathways and numbers indicate relative % representation. Bar charts represent secondary metabolic pathways (top 3 are shown). A: Carbohydrate metabolism; B: Amino acid metabolism; C: Metabolism of cofactors and vitamins; D: Signal transduction; E: Membrane transport; F: Cell growth and death; G: Cell motility; H: Cellular community prokaryotes; I: Infectious diseases; J: Neurodegenerative diseases; K: Cancers; L: Replication and repair; M: Translation; N: Folding, sorting, and degradation; O: Environmental adaptation; P: Endocrine system; Q: Immune system.