High Prevalence of Antimicrobial Resistance and Integron Gene Cassettes in Multi-Drug-Resistant Klebsiella pneumoniae Isolates From Captive Giant Pandas (Ailuropoda melanoleuca)

Abstract

Multi-drug-resistant Klebsiella pneumoniae (MDR K. pneumonia) is increasingly being reported with corresponding increase in morbidity and mortality all over the world. However, limited information is available concerning MDR K. pneumonia in giant pandas. The objective of this study was to grasp the drug resistance profile of MDR K. pneumonia isolated from giant pandas. A total of 182 K. pneumoniae isolates were collected from fresh feces of 94 captive giant pandas of different ages and sex and separated by season. We performed a standard disk diffusion antimicrobial susceptibility test with the isolates and further evaluated the antibiotic resistance genes (ARGs) of multi-drug-resistant strains by high-throughput quantitative PCR. In addition, we then analyzed mobile genetic elements (MGEs), integron gene cassettes, and the multi-locus sequence typing of multi-drug-resistant strains by PCR. Antimicrobial susceptibility testing results demonstrated that a total of 30 (16.5%) K. pneumoniae isolates showed multiple drug resistances. The thirty MDR K. pneumonia isolates were mainly resistant to amoxicillin (100.0%), doxycycline (86.7%), chloramphenicol (60.0%), compound trimethoprim (60.0%) and trimethoprim (56.7%). Fifty different types of antibiotic resistance genes were found, which included a total of 671 antibiotic resistance genes, in the 30 multi-drug-resistant isolates. The top ten resistance genes were: vanTC-02, aacC, blaCTX-M-04, blaSHV-01, blaSHV-02, ampC-04, blaOXY, tetD, blaTEM and tetA-02. Thirteen mobile genetic elements were detected, of which IS26 (96.67%) and intI1 (96.67%) had the highest frequency. The thirty MDR K. pneumonia isolates were negative for the traA, traF, tnsA, IS1133, ISpa7, ISkpn6, intI2 and intI3 genes. Moreover, a further investigation of integrons revealed that two types of specific gene cassettes (dfrA12 + orfF + aadA2 and dfrA12 + orfF) were identified in class 1 integrons. Multi-locus sequence typing results showed that 22 STs in the thirty MDR K. pneumonia isolates were identified, the main type was ST37 (5/30). Our results illustrate that effective surveillance and strict biosecurity strategies should be taken to prevent the spread of multi-drug-resistant bacteria, and monitor the emergence of mobile genetic elements and integrons.

Prevalence of antimicrobial resistance and virulence genes in Klebsiella pneumoniae and Congenetic Raoultella Isolates from captive giant pandas

“…In 2020, Yan X et al obtained the first isolates of MDR-KP in 16.5% (30/182) from the feces of healthy captive giant pandas [ 37 ]. In the present study, 42 K .…”

“…While some studies have claimed that chloramphenicol and tetracycline have not been used as therapeutic drugs in giant pandas [ 16 , 37 ], Zhang AY et al found that 13.56% of the bacterial strains isolated from the feces of 30 giant pandas were resistant to chloramphenicol and tetracycline [ 16 ]. Yan X et al found resistance rates of 30 MDR-KP strains to chloramphenicol and tetracycline of 60% [ 37 ]. Interestingly, the MDR strains found in this study showed slightly higher resistance to chloramphenicol (70%) and trimethoprim-sulfamethoxazole (90%).…”

Detection of Klebsiella pneumonia DNA and ESBL positive strains by PCR-based CRISPR-LbCas12a system

“…A recent discovery reveals that 73.1% of K. pneumonia are resistant to at least one antibiotic ( Petrosillo et al, 2019 ; Sharahi et al, 2021 ). Multi-drug resistant and extensively drug-resistant K. pneumonia strains are increasing ( Peng et al, 2020 ; Yan et al, 2021 ; Yang et al, 2021 ). Effectively controlling K. pneumonia requires the direct knowledge of drug-resistant information ( Ludden et al, 2020 ).…”

Antimicrobial resistance, serogroups, virulence gene profiles and MLST of Escherichia coli from giant panda

“…The antibiotic susceptibility testing results were interpreted according to Clinical and Laboratory Standards Institute (2020) , with E. coli ATCC25922 used as a control and quality assurance measure. All the antibiotics used in this study were based from recent publications on the microbial resistance of giant pandas and recommendations from CRBGP veterinarians ( Zhang et al, 2009 ; Guo et al, 2015 ; Chen et al, 2018 ; Zou et al, 2018 ; Zhu et al, 2020 ; Yan et al, 2021 ). Multidrug resistance (MDR) was determined if there was resistance to three or more classes of antibiotics ( Magiorakos et al, 2012 ; Zhu et al, 2020 ).…”

“…The abundance of ARGs in different E. coli isolates was evaluated through high-throughput quantitative PCR (HT-qPCR) of ARGs performed by Yearth Biotech (Changsha, China) using the SmartChip Real-time PCR (Warfergen Inc., United States) ( Yan et al, 2021 ). In total, 47 primer sets (including a 16S rRNA gene primer sets) were used ( Table 1 ).…”