| Staphylococcus spp.is an opportunistic pathogen that is widely found in humans,animals,and the environment.Staphylococcus of animal can also enter the digestive tract and respiratory tract by swallowing,snubbing,or wounds.It is a systemic disease caused by blood.A large number of staphylococci can also be isolated in oral diseases such as rickets,periodontitis,and pulpitis.At present,there have been partial reports at home and abroad that more staphylococci have been isolated from the giant panda's animal house,the respiratory tract of fatal pneumonia and the abnormal tooth.However,there is no report on the drug resistance of Staphylococcus.Therefore,we aimed to explore the species of culturable bacteria and the antibiotic resistance of Staphylococcus spp.in the oral cavity of the giant panda.In the experiment,15 juvenile and adult giant pandas were selected,and 60 oral saliva samples(2 each)were collected.After isolation and purification of the bacteria,the oral culturable species were identified based on the 16 s rRNA gene.Then,the most suspected staphylococci isolated from the oral bacteria of giant panda were selected,and the species of staphylococcus were further identified by Gram's microscopic examination,biochemical identification and coagulase test.K-B paper method and multiplex PCR were used to study the resistance of Staphylococcus to 12 antibiotics and the carrying status of 12drug-resistant genes.The results showed that 302 culturable bacteria were isolated from the giant panda samples,mainly belonging to 78 species of 31 genera,mainly Gram-positive bacteria,including Staphylococcus,Streptococcus,Pseudomonas and Rothia,Enterobacter,Bacillus,Sphingosine bacillus genus,Acinetobacter,klebsiella to advantage bacterium group.Among them,80 coagulase-negative staphylococci(CoNS)(39 juvenile,41 adult)were identified,and coagulase-positive staphylococci were not isolated.78.75% of Staphylococcus is resistant to at least one antibiotic,more than half of the strains are multi-drug resistant,and the five-drug resistance rate is 6.25%.The juvenile giant panda Staphylococcus is mainly resistant to 2 kinds of antibiotics(28.21%),and the adult giant panda Staphylococcus is mainly resistant to 3 kinds of antibiotics(31.71%).Among them,vancomycin has the highest resistance rate(68.75%),followed by azithromycin(51.25%),and the aminoglycoside and tetracycline resistance were less than 20%.Only the juvenile giant panda Staphylococcus were sensitive to ciprofloxacin and chloramphenicol.At the same time,48.75% of methicillin-resistant coagulase-negative staphylococci(MRCNS)were isolated.By comparison,the resistance of MRCNS was much higher than that of No-MRCNS.Among the 80 strains of Staphylococcus resistance genes,68.75% of the strains isolated at least one resistance gene,with the highest separation rate of BlaZ gene(41.25%),followed by mecA(40.00%),cfr(38.75%),VanB(25.00%).80 strains of CoNS carried vancomycin,?-lactam,aminoglycosides,macrolides,tetracyclines,chloramphenicol resistance genes in VanB;BlaZ/mecA;aacA-aphD;Erma/ermb;tetK;cfr gene-based.The juvenile giant panda staphylococcus mainly carries 1-2 resistant genes(82.61%);the adult giant panda staphylococcus mainly carries 2-4 resistant genes(75.00%),and the two of them carry the highest 6 resistant genes..In summary,80 strains of Staphylococcus have extensive drug resistance,among which MRCNS has higher drug resistance,and the resistance of adult giant panda Staphylococcus and the rate of drug-resistant genes are higher than that of juvenile giant pandas,proving the oral cavity of giant pandas.Staphylococcus spp has a certain degree of drug resistance,so it is necessary to strengthen the resistance monitoring of giant pandas and their living environment,control and reduce the generation and spread of drug-resistant strains. |
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