Study On The Influence Of The Chinese Traditional Compound Medicine "LH" On Drug Gene StrA,SulⅡ Of Pasteurella

Pasteurellosis mainly by specific serotypes of Pasteurella multocida as a deadly infectious diseases cause of livestock and poultry were suffering.And the high morbidity and mortality, often caused to the aquaculture more serious losses. for this, a large number of antimicrobial drugs used in clinical to control this disease, caused widespread Pasteurella resistance, especially in high-resistant strains has led to a majority of the clinical effect of drugs on the lower resistant or ineffective. So we focused on this characteristic of it has a very high resistance to streptomycin and sulfa drug,and researched the lianhuang of Chinese herbal formula to eliminate Pasteurella for resistance of streptomycin and sulfa drug,and discussed it from the level of molecular biology on lianhuang of Chinese herbal formula on the Pasteurella strA resistance gene Sulâ…¡and the mechanism of influenceThis study was preliminary in vitro tests to research Chinese medicine "lianhuang" on resistance drug of Pasteurella's sulfa and aminoglycoside resistance in vitro. Point species by agar plate culture method detected sub-MIC of Chinese herbal formula "lianhuang" on sulfa, aminoglycosides inhibit the rate of drug resistance of Pasteurella, while "lianhuang" resistant strains of the Pasteurella Pasteurella inhibition rate compared with sensitivity by statistical analysis to determine effect of inhibitors of drug resistance.According to Pasteurella streptomycin resistance gene and sulfonamide resistance gene Sulâ…¡StrA sequence synthesis 2 pairs of primers, which isolates of Pasteurella multocida of avian origin plasmid DNA as a template drug-resistant strains in clinical,by PCR, amplified Papanicolaou Bacillus StrA, Sulâ…¡gene. To about 1136bp of StrA gene and 796 bp of Sulâ…¡cloned into pMD-18T vector and sequenced. Compared and analyzed the resistance gene of Before and after the experiment on the Chinese medicine acted in Pasteurella (P0, P1, DH01, DL01, P2). detected the effect of Chinese medicine "lianhuang" on the Pasteurella resistance gene StrA, SulII. Preliminary discussed Chinese Medicine "lianhuang" reduce Pasteurella on sulfonamides, aminoglycoside resistance mechanisms.Results:Species Act by the agar plate results point known, Chinese medicine "lianhuang" on the resistance of Pasteurella PI, DL01, DH01 and Mingan Jun inhibition rate was no significant difference, between PO and amikacin on multidrug resistance of Pasteurella DL01, P2 inhibition rate and significant difference between P0, streptomycin resistant Pasteurella P2 on inhibition rate compared with PO significant difference. The results from the sensitivity disk method known by the Chinese medicine "lianhuang" effect, all of the antibiotics between on the resistance of Pasteurella P1, DL01, DH01 size and PO inhibition ring has no significant difference, between sulfonamides on the suppression P2 Ring size and strain PO was significant difference, between streptomycin and amikacin on the size of P2 and inhibition zone PO was significant difference; and between amikacin on the DL01 inhibition ring size and PO was significant difference. Resistance by cloning and sequencing Pasteurella StrA (PI) gene sequences published in GenBank sequence (NC₀01774) homology was 99.7%, StrA(DLOl) gene sequences published in GenBank sequence (NC₀01774) homology was 99.0%of its StrA gene sequences have two base mutation that 204C→G,686T→C resistance in Pasteurella P1 through cloning and sequencing of SulII gene sequences published in GenBank (NC₀01774) homology of 99.5%DH01 Sulâ…¡gene sequences published in GenBank (NC₀01774) 98.7%homology. Chinese medicine acted before and after the Pasteurella SulII gene in the 580 to 600 base pairs in the region has changed, but the corresponding amino acid has not changed.Conclusion:Chinese medicine "lianghuang" can effectively reduce the Pasteurella on sulfonamides, streptomycin resistance; its mechanism is due to change part of their resistance genes StrA, SulII, so the resistance drug gene inactivation.