Effect of plant Lactobacillus P-8 on the gut flora of chickens using lincomycin.

.The growth of broiler chickens is closely related to their intestinal health, and the farming industry often uses antibiotics to control the occurrence of poultry diseases and improve their survival. However, prolonged use of antibiotics can lead to intestinal malnutrition, diarrhoea and slower growth rates in broiler chickens. Plant Lactobacillus P-8 as an excellent probiotic lactic acid bacteria has been shown to regulate the intestinal flora of a variety of animals, in order to further study the effect of micro-ecological preparations to replace antibiotics, this experiment explores the plant Lactobacillus P-8 on the use of lincomycin chicken intestinal flora improvement.Experimental DesignExperimental Details 1. Experimental Subjects Selected 1-day-old broiler chicken for a 30-day animal experiment, randomly divided into probiotic group (QD1-5) and control group (QD6-10).. 2. The experimentalbasic diet is shown in Table 1, and all broiler diets are added to the diet of lincomycin hydrochloric acid. Plant Lactobacillus P-8 (2x10
6
CFU/ml)
also added to the probiotic group.. 3. Testing methodthe 30th day of chicken faeces samples to test the structure of the intestinal bacteria, related metabolic characteristics, etc. (Figure 1).. Figure 1: Details of the experimentDesign Table 1: Basics of the chicken dietResults 1. DNA sequencing coverage and intestinal bacteriobi diversity DNA sequencing of the feces samples of the broiler was obtained with a total of 152.7Gb of high-quality data, with an average of 310,214 sequences of 16S rRNA per bacterium and 31,528 OTU identified. As the sequencing depth increases, the sparse curve of the Shannon Index levels off, suggesting that while it is possible to detect new OTUs, the sequencing depth has met the demand and the vast majority of microorganisms can be captured.compared to the control group, only the probiotic group had a super 1 index greater than the control group, while the Shannon index, the Simpson index, and the number of species observed were significantly lower than the control group (P<0.05; Table 2). This indicates a significant difference in the diversity of gut bacteria before and after treatment, in which the diversity of intestinal flora decreases after the intervention of Bacillus plant P-8.. Table 2: Details of the diversity of intestinal bacteria in broilers 2. Structure of the intestinal bacteria The composition of the intestinal bacteria at the gate and genus levels of each sample is shown in Figure 1, mainly thick-walled bacillus (63.58%), metamorphic bacteria (17.31%) and psylobacteria (10.87%)(Figure 1a). At the genus level, a total of 885 genus were identified, of which 13 were more than 1 per cent of the total sequence (Figure 1b). These 13 genus account for 58.34 percent of the gut microbiome, of which lactic acid bacteria are the most common genus.Although there was no significant difference between the relative abundance of lactic acid bacteria between the probiotic group and the control group, the proportion of lactic acid bacteria in the probiotic group (17.64%) was slightly higher than that in the control group (15.74%).. Figure 2: Intestinal bacterial structure of broilers, a at the gate level, b at the genus levelbased on unstialized UniFrac distance main coordinate analysis (PCoA) shows that there are different microbiome structures between the probiotic group and the control group (Figure 3) a), however, from the Weighted UniFrac Distance Primary Coordinate Analysis (PCoA), it is shown that there is a slight overlap between the probiotic and control groups (Figure 3b), and this slight difference may indicate that the microbiome structure differences between the two groups are in the lineage with the least OTU.. Figure 3: UniFrac Distance Main Coordinate Analysis (PCoA) We also compared the relative abundance of the probiotic group and the control group of bacterial genus, where the probiotic group
Gallicola, Helcococcus, Dorea The number of Desemzia
and
Wautersiella
decreased significantly, while the trend of
Oceanimonas
and
Flavobacterium
was reversed. . 3. Gene function notes for intestinal bacteria A total of 3,979 COGs, or 45.11% of all genes, were identified, and these COGs fall into 23 functional categories. In all samples, the L (replication, duplication and repair), E (amino acid transport and metabolism) and G (carbohydrate transport and metabolism) categories dominated, while more gene sequences were found in the probiotic group, namely P (inorge ion transport and metabolism), C (energy generation and transformation), N (cell movement) and A (RNA processing and modification) (Figure 4). This indicates that after the intervention of plant Lactobacillus P-8, there are more clusters of the same origin groups classified as P, C, N and A at the macro genome level. . Figure 4: Notes on gene function PCA analysis was conducted based on the distribution of COG in order to assess the overall differences in intestinal bacterial function genes between probiotic and control groups. In the PCA analysis diagram, a significant separation between the two groups was observed. This suggests that the treatment of Plant Lactobacillus P-8 led to an overall change in functional genes. . 4. Notes on metabolic pathways In samples of intestinal flora in the probiotic group, it was found that the metabolic pathways involved in whiplash assembly, bacterial trending, bacterial secretion system, lipid polysaccharide biosynthesis, and nitrogen metabolism, sulfur metabolism and vitamin biosynthesis were more obvious. The probiotic group was significantly less involved in semi-lactose degradation, carbon fixation, multiple sugar operating systems, and metabolic modules associated with ucose glycosomes than in the control group. In the PCA analysis diagram, a significant separation between the two groups was observed. This indicates that the probiotic group has more metabolic modules with high metabolic activity. results plant Lactobacillus P-8 can improve the intestinal flora of chickens using lincomycin, and the diversity of intestinal flora decreases. Effectively increased the abundance of lactic acid bacteria and beneficial bacteria, while reducing the abundance of pathogenic bacteria. At the macrogenomic level, Plant Lactobacillus P-8 enhances functional classification of homogenous clusters related to inororated ion transport and metabolism, energy generation and transformation, cell movement, and RNA processing and modification. In terms of metabolic pathways, the intake of lactobacillus P-8 in plants led to increased gene abundance associated with whiplash assembly, bacterial trending, nitrogen metabolism, sulfur metabolism, and vitamin biosynthesis.
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