| In recent years, with the development of economy, the fur animal breeding industry in Shandong province has developed very rapidly.At present, Shandong province has become the first province of China’s fur animal breeding, fur animal husbandry is now about 1/2 of the country’s total.However, with the gradual increase of the degree of animal husbandry in our province, the incidence of fur animal is also rising.This has become an important factor restricting the development of China’s fur animal industry, which caused great economic losses to the fur animal breeding industry.Mink hemorrhagic pneumonia is one of the important diseases seriously hinder the mink industry, the epidemic is more serious.Research shows that the canine distemper virus, Influenza, Pseudomonas aeruginosa, Klebsiella peneumoniae1, Staphylococcus, Escherichia Coli are common pathogens causing mink hemorrhagic pneumonia.This study uses conventional bacterial isolation and identification technology, isolation and identification of the bacterial pneumonia in mink lung tissue of 109 patients with hemorrhage.Among them, there were 10 strains which could not be purified, but 99 strains could be cultured in pure culture,Among them, 99 strains of bacteria showed different growth forms on the common nutrient agar medium.After the extraction of bacterial genomic DNA, 16 SrRNA gene sequences were obtained by PCR method using the r RNA 16 S sequence of the bacteria and the primers 16S-1492 R and 16S-27 F were amplified and sequenced.Using NCBI-BLAST software, the sequencing sequence in the bank Gene database for homology search analysis, preliminary determination of species of bacteria, and then according to the characteristics of bacteria in biochemical reactions to determine the characteristics of bacterial species.The experimental results show that,the 99 strains of bacteria, which 31 strains of Klebsiella pneumoniae bacteria, the bacteria detection rate was 28.4%(31/109); 13 strains of Pseudomonas aeruginosa, the bacteria detection rate of 11.9%(13/109); 34 strains of Escherichia coli, and the bacteria detection rate was 31.2% 34/109; 8 strains of staphylococci, the bacteria detection rate of 7.3%(8/109); 2 strains of Proteus, 3 strains of Enterococcus faecalis, 2 strains of Proteus mirabilis, two strains of Bacillus sp., 1 strain of Citrobacter and 1 strain of Citrobacter acid bacilli, 1 strain of Bacillus cereus, 1 strain of orange tiny coli.The results of homology analysis of 16 SrRNA sequence showed that,the 13 strains of bacteria isolated from the nucleotide sequence homology was 84.2%-100%, which, in addition to SD P.a-9 and other 12 strains of low homology between 84.2%-87.3% and other 12 strains homology between the relatively high. Genetic evolution analysis showed that P.A-9 SD was in the first branch with the other 12 strains. P.a-4 SD and P.a-12 SD were closely related, P.a-1 DM5 and the reference strain SD, s7ps5 and the same small branch on the close relationship. The nucleotide sequences of the 31 isolates were 75.2%-100%, Kp SD 8 and the other 30 strains, which were 75.2%-76.6%. The other 30 strains were highly homologous. Genetic evolution analysis showed that Kp SD 8 and the rest of the 30 strains of the more distant, in the first branch.The nucleotide sequence homology between the 34 strains isolated from the isolates was 96.3%-100%, and the homology was very high. Genetic evolution analysis showed that Eco-4 SD and the other 33 strains in the evolutionary relationship is far, in the first branch, the other 33 strains in the second branch, the genetic relationship is relatively close. The nucleotide sequences of the 8 isolates were 97.3%-100%, and the homology between Sau-5 SD and Sau-6 SD was higher, which was in a small branch. But sau5, sau6 and other six strains with low homology. SD sau1, SD sau2, SD sau3, SD sau4, SD sau7 and sau8 among the six isolates were highly homologous, were more than 98%. Analysis results show that the evolutionary relationship of SD sau5 and SD sau6 were closely related in the first branch of genetic evolution. Close to SD sau2 and SD sau3 relationship, near SD and sau4 relationship, near sau7 and SD SD sau8 relationship. SD sau1, SD sau2, SD sau3, SD sau4, SD sau7 and sau8 six strains in phylogenetic tree of the second branch.Drug sensitivity test used to detect strains of Klebsiella pneumoniae, found the bacteria to florfenicol, Amikacin, ceftazidime, levofloxacin and drug sensitivity; through drug sensitivity test for the detection of strains of Pseudomonas aeruginosa, it is found that the bacteria to cefoperazone, streptomycin, Amikacin, ceftazidime, levofloxacin and drug sensitivity; the drug sensitivity test for the detection of Escherichia coli, found the bacteria to ceftriaxone, cefotaxime, ceftazidime, Amikacin, florfenicol and drug sensitivity; through drug sensitivity test for the detection of Staphylococcus aureus strains, it was found that the bacterium of amoxicillin, cefotaxime, cefoperazone, ampicillin, florfenicol etc. drug sensitive.The disease was dissected and found mainly in the lung hemorrhage.Of disease inspection materials for molecular biology detection. The main pathogenic bacteria of pneumonia in mink analysis show grams Klebsiella, Pseudomonas aeruginosa, bacillus, Escherichia coli and Staphylococcus aureus infection rate is relatively high, most to amikacin, cefotaxime, drug sensitive, can be symptomatic treatment according to the illness.This experiment preliminary understanding bleeding pneumonia pathogen infection rate of Shandong Province mink, understanding mink hemorrhagic pneumonia bacterial disease pathogens and drug resistance, for further prevention and treatment of mink hemorrhagic pneumonia and provide the basis. |
