Diagnosis And Controlling With Traditional Chinese Medicine And Antibiotic On Hafnia Alvei Disease From Apis Cerana Fabricius

Objective: To accurately diagnose the bacterial diseases of Apis Cerana Fabricius in a bee farm in Guizhou Province,and to Screen Chinese and Western drugs sensitive to pathogenic bacteria and to provide the corresponding treatment guidelines for the Chinese bee disease caused by this strain of bacteria.Methods:1)After isolation and purification of bacteria,morphological observation and biochemical experiment,a pathogen was obtained.The 16 s r RNA gene of the bacteria was sequenced by universal primers,and the species of the strain was finally determined.2)The antimicrobial resistance of the isolated strain was detected by the method of drug sensitive tablets,and the antibiotic drugs and the single Chinese herbal medicine which had the inhibitory effect on the strain were selected.3)The MIC values of the bacterial isolate for the selection of antibiotics and single Chinese medicine were determined by double dilution method with micro broth.4)The combined bacteriostatic effect of Chinese and western medicine was determined by using chessboard method and the bacteriostatic concentration index(FICI)was calculated.5)The best group after the combination of experimental antibiotics and single Chinese herbs was selected to carry out animal experiments to verify the effect.Results:1)The pure culture of 6 bacteria was isolated and detected in the disease.One of them was identified by pathogenicity experiment.The pure culture of H1 bacteria was identified as pathogenic bacteria,and the pure culture of this strain was used as the hive.2)The bacteriostatic effect of polyploid seed and Coptis chinensis is the most obvious.Scutellaria baicalensis,Hawthorn,leaves of Artemisia and Forsythia Forsythia also have strong bacteriostatic effect.Bupleurum chinense and andrographis andrographis had weak bacteriostatic effect,but the leaves of Daqingye,Houttuynia cordata,mint and dandelion lacked bacteriostasis.The bacteria were resistant to penicillin,clindamycin,erythromycin,minocycline,oxacillin and cefalexin in Western drug susceptibility tests;for maximycin,tetracycline and doxycycline were moderately sensitive;for gentamicin,furazolidone,Compound sulfamide,ofloxacin,norfloxacin,ciprofloxacin,ampicillin,cefuroxime,ceftazidime and cefoperazone are sensitive.Finally,Chinese medicine was selected as gallnut,Coptis chinensis,Scutellaria baicalensis,Forsythia forsythia,Artemisia argyi and Hawthorn,and the western medicine was gentamicin sulfate and ampicillin sodium for MIC and FICI detection.3)The minimum inhibitory concentration of Galla chinensis decoction alone was 3.9mg / ml,that of Coptis chinensis decoction alone was 7.8 mg / ml,and that of Scutellaria baicalensis decoction was 15.6mg / ml.The minimum inhibitory concentration(MEC)of Artemisia argyrifolia decoction was 31.2 mg / ml,that of Forsythia suspensa decoction was 31.2 mg / ml,and that of Hawthorn decoction was 62.5 mg / ml.The minimum inhibitory concentration of western medicine was 2.5 ? g / ml for gentamicin sulfate alone and 20 ? g / ml for ampicillin sodium alone.4)After the combination of Chinese and western medicine,the effect was better.The synergistic combination was Gentamycin Sulfate injection and decoction of Galla chinensis,Coptis chinensis,Radix Scutellariae,ampicillin sodium and Coptis chinensis and Radix Scutellariae.The best combination was gentamicin sulfate and Coptis chinensis decoction,and its FICI value was 0.18.The drug combinations in other groups were additive and irrelevant.Gentamycin sulfate and Coptis chinensis decoction were used in animal experiments,the results showed that Gentamycin sulfate and Coptis chinensis decoction had a good curative effect on the infectious diseasesConclusions:1)The results of isolation and identification of pathogenic bacteria and pathogenicity test showed that the infection is caused by Hafnia alvei in a bee farm in Guizhou province.2)The strain was only resistant to a small number of antimicrobial agents,indicating that the bacteria had not been prevalent in the colony for a long time,and that the colony had no multidrug resistance caused by drug abuse.3)A variety of antibiotics,such as ofloxacin,ciprofloxacin,cefadine and ceftazidime,can be obtained by the bacteriostasis test in vitro,and the decoction of Chinese medicine of Chinese gallnut and Rhizoma Coptidis can also be used to suppress the bacteria effectively.3)In vitro bacteriostatic test showed that gentamicin combined with Coptis chinensis decoction had a good bacteriostasis effect against Hafni's bacteria in honeycomb.