Effects Of Antibiotics On Phosphate Accumulating Organisms And Reversal Of Cellular Substances Of Phosphate Accumulating Organisms By Chinese Medicine

In recent years,antibiotics as an emerging pollutant,have gradually attracted people’s attention due to their serious danger to water pollution.Gentamicin,which can exist stably in water,can lead to intracellular leakage of proteins and DNA in the cell due to its personality of interfering with protein synthesis.As a result,the normal function of microorganism was seriously blocked in the process of sewage biological treatment.Moreover,with the accumulation of antibiotics in the process of biological phosphorus removal in sewage,the resistance genes of polyphosphate accumulating organisms are induced.Moreover,the damage to the environment and human body caused by excess sludge discharge and other ways is beyond calculation.Therefore,to find ways to eliminate or reverse the intracellular resistance genes produced by gentamicin-stressed polyphosphate accumulating organismsis of great significance.Using traditional methods to eliminate intracellular resistance genes has the disadvantages of easy destruction of bacteria structure,high cost and the effect is not obvious.However,there are few researches suggest that the way of using Chinese medicine to eliminate or reverse the intracellular resistance genes of microorganisms is of low toxicity and high efficiency.However,there are few researches focus on the effect of gentamicin on polyphosphate accumulating organisms and the reversal of cellular substance of polyphosphate accumulating organisms by traditional Chinese medicine.In this paper,polyphosphate accumulating organisms was cultured by alternating anaerobic/aerobic plate iteration.Combining5-bromine-4-chloro-3-indole-3-phosphate solution（BCIP）blue white spot staining selection results and phosphorus removal efficiency test results to screen polyphosphate accumulating organisms.Gentamicin is added to it.Through comprehensive analysis of intracellular protein,DNA leakage and changes in conductivity of bacterial liquid and phosphorus removal performance test,the cell membrane and functional integrity of polyphosphate accumulating organisms are analyzed.The effect of Traditional Chinese medicine on the protein structure before and after the reversal was analyzed by Raman spectroscopy.Finally,fluorescence quantitative PCR was used to detect the changes of intracellular gentamicin resistance gene abundance before and after traditional Chinese medicine treated.The main experimental results are as follows:（1）Using an SBR reactor to enrich polyphosphate accumulating organisms by adopting anaerobic/aerobic alternation mode.BCIP blue spot screening,poly--hydroxybutyric acid（PHB）,heterochromic particle staining and gene sequencing were used to identify the selected polyphosphate accumulating organisms.The identification results suggested that:No.1 strain was Exiguobacterium sp.ZWU0009,No.2 strain was Bacillus sp,No.4 strain was Acetobacter Acetobacter strain SI17,No.5 strain was Acetobacter Acetobacter strain B.No.1 Exiguobacterium sp.ZWU0009 and No.4 Acetobacter Acetobacter strain SI17 were selected as the following test strains.（2）Take exiguobacterium sp.zwu0009 and Acetobacter Acetobacter strain as target strains.After gentamicin acts on strains and acquires resistance genes,three traditional Chinese medicines,which respectively are Scutellaria baicalensis,Galla chinensis and Artemisia argyis are used to act on exiguobacterium sp.zwu0009 and Acetobacter Acetobacter strain.The leakage of protein and DNA,conductivity of bacteria solution and phosphorus removal efficiency of polyphosphate accumulating organisms were determined before and after the treatment of three Chinese medicine.The test results suggests that:After Scutellaria baicalensis,Galla chinensis and Folium artemisiae argyi acted on gentamicin-resistant exiguobacterium sp.zwu0009 and Acetobacter Acetobacter strain,the difference of cell membrane index of the leakage of protein and DNA,cell conductivity and phosphorus removal efficiency between the two strains was smaller than that between control samples.It indicates the traditional Chinese medicine has a better reversal effect on Exiguobacterium sp.ZWU0009 and Acetobacter Acetobacter strain.（3）Two kinds of polyphosphate accumulating organisms,exiguobacterium sp.zwu0009 and Acetobacter Acetobacter strain,were selected as the strain samples.After the exacts of Scutellaria baicalensis,Galla chinensis and Folium artemisiae argyi acted on gentamicin-resistant exiguobacterium sp.zwu0009 and Acetobacter Acetobacter strain,a protein extraction kit is used to extract the total protein in the bacterial cells.The extracted protein was scanned and analyzed by Raman spectrometer.The results indicates,the peaks of protein Raman spectrum of resistant Acetobacter and Micrococcus are generally around 860 cm^-1,918 cm^-1,942cm^-1,1000 cm^-1,1025 cm^-11 and 1002 cm^-1.The corresponding bond positions have the characteristics of high bond energy,long bond length and strong stability.After Scutellaria baicalensis,Galla chinensis and Folium artemisiae argyi acted on gentamicin-resistant Exiguobacterium sp.ZWU0009 and Acetobacter Acetobacter strain,the protein of resistant strains was transformed into the protein of the same peak position in the control group.（4）Three traditional Chinese medicines of Scutellaria baicalensis,Galla chinensis and Folium artemisiae argyi are used to act on gentamicin-resistant Exiguobacterium sp.ZWU0009 and Acetobacter Acetobacter strain.The gentamicin resistance gene abundanceswas determined by fluorescence quantitative PCR.The test results suggest that,the gentamicin resistance gene abundances of resistant Acetobacter and resistant microbacteria were higher than those of the control group.After the reversal of the three traditional Chinese medicines of Scutellaria baicalensis,Galla Chinensis and Folium artemisiae argyi,gentamicin gene abundance of gentamicin-resistant Exiguobacterium sp.ZWU0009 and Acetobacter Acetobacter strain decreased significantly and returned to the level of control group.Scutellaria baicalensis,Galla chinensis and Folium artemisiae argyi effectively reversed gentamicin resistance genes in polyphosphate accumulating organisms,which provides a new idea for the removal of resistance genes in the cell environment.This research has certain practical significance and guiding value of basic theory.Figure[26]Table[16]Reference[104]...