Analysis On Cutaneous Bacterial Community Of Rana Dybowskii And Effect Of Medicated Bath

The Rana dybowskii is one of the dominant species of amphibians in northeast China and is also a valuable economic frog species.Rana dybowskii is susceptible to disease.For the breeding industry,the prevention and treatment of disease is a key factor in the success of breeding.Rana dybowskii have a large number of bacteria on their skin.These bacteria are closely related to the health of Rana dybowskii.In this experiment,the high-throughput sequencing technology was used to detect the bacterial communities structure of wild and farmed Rana dybowskii skin in order to understand the species and abundance of bacterial communities and the differences of bacterial communities and Potential pathogenic genera between the two groups of Rana dybowskii in different environments.For the Rana dybowskii,it is difficult to take the administration methods such as injections and feedings after illness.Due to the special structure and function of Rana dybowskii skin,the medicated bath is selected.In this experiment,a medicated bath was made on the Rana dybowskii to explore the cutaneous bacterial community structure of the Rana dybowskii after the medicated bath,and then to understand the effect of the medicated bath on the cutaneous bacterial community structure and Potential pathogenic genera.Experiments were conducted to detect and analyze the cutaneous bacterial community structure of wild and farmed Rana dybowskii(wild group n=7,farmed group n=7);and the effect of medicated bath on the cutaneous bacterial community structure of farmed Rana dybowskii include antibiotics(gentamicin),probiotics(Bacillus subtilis)and traditional chinese medicine(rhubarb powder decoction)(control group n=7,antibiotic group n=14,probiotics group n=7,traditional chinese medicine group n=7).The skin samples of the wild and farmed Rana dybowskii and the skin samples after different medicated baths were taken,and the species composition and abundance informations of the bacteria communities were obtained after sequencing and analysis.Through the Alpha diversity analysis,Beta diversity analysis and the comparison of potential pathogenic genera,the differences in bacteria community diversity and structure and potential pathogenic genera in each group were examined and compared.The results are as follows:(1)Alpha diversity and Beta diversity in wild and farmed groups are significantly different.The dominant phyla of the farmed group are Firmicutes,Proteobacteria,Actinobacteria,Bacteroidetes.The dominant phyla of the wild group are Proteobacteria,Bacteroidetes,Firmicutes.Seven potential pathogenic genera were analyzed.Citrobacter,Proteus and Staphylococcus in the farmed group were significantly higher than those in the wild group;Pseudomonas in the wild group was significantly higher than that in the farmed group.Aeromonas,Chryseobacterium,Streptococcus were not significantly different in the two groups.(2)The antibiotics bath had no significant effect on the Alpha diversity of bacteria community but had a significant effect on Beta diversity.The antibiotic bath had no significant effect on the diversity of the Alpha.After the bath,the dominant phyla are Proteobacteria,Bacteroidetes,Actinobacteria,etc;the dominant genera are Arthrobacter,Elizabethkingia,Pseudomonas,etc.Antibiotics bath has significant inhibitory effects on Aeromonas,Chryseobacterium.(3)The Probiotics bath had no significant effect on the Alpha diversity of bacteria community but had a significant effect on Beta diversity.After the bath,the dominant phyla are Proteobacteria,Bacteroidetes,Firmicutes,etc;the dominant genera are Elizabethkingia,Chryseobacterium Arthrobacter,etc.Probiotics bath has significant inhibitory effects on Aeromonas.(4)The traditional chinese medicine bath had no significant effect on the Alpha diversity of bacteria community but had a significant effect on Beta diversity.After the bath,the dominant phyla are Bacteroidetes,Proteobacteria,Actinobacteria,etc;the dominant genera are Elizabethkingia,Arthrobacter,Flavobacterium,etc.Traditional chinese medicine bath has significant inhibitory effects on Aeromonas.(5)A comprehensive comparison and analysis was conducted on the results of medicated bath.The results of ANOSIM analysis showed that there were significant differences between antibiotics group and the other three groups,and there was a significant difference between probiotics group and control group,traditional chinese medicine group and control group.There was no significant difference between probiotics group and traditional chinese medicine group.The experiment concluded that there was a significant difference in the bacteria community between two groups.The main dominant phyla of farmed group include Firmicutes,Proteobacteria,Actinobacteria,Chloroflexi,Fusobacterium,Bacteroidetes and Cyanobacteria;the main dominant phyla of wild groups include Proteobacteria,Bacteroidetes,Firmicutes and Actinobacteria.The gentamicin,Bacillus subtilis and rhubarb powder decoction medicated baths had significant effects on the Beta diversity of the cutaneous bacterial communities and inhibited several potential pathogenic genera.This study provided basic data for the research and conservation of Rana dybowskii and provided a theoretical basis for the selection of drugs.