Antibiotic induced biofilm formation of novel multidrug resistant Acinetobacter baumannii ST2121 clone

Author(s):  
Müberra Çimen ◽  
Azer Özad Düzgün

AbstractThe aim of this study was to identify antimicrobial resistance and virulence factor genes exhibited by multidrug resistant (MDR) Acinetobacter baumannii, to analyze biofilm formation and to investigate clonal subtypes of isolate. Whole genome sequencing was done by Illumina NovaSeq 6,000 platform and multilocus sequence typing (MLST) was performed by Oxford and Pasteur typing schemes. Influence of imipenem and levofloxacin on biofilm formation was investigated in 96-well plates at 3 replicates. The strain was found to carry OXA-23, OXA-51-like, AmpC and TEM-1 beta-lactamases. The sequence of the blaOXA-51-like gene has been identified as a blaOXA-66. According to Pasteur MLST scheme the strain displayed ST2 allelic profile. However, based on Oxford MLST scheme this strain represents the new ST2121, as the gdhB gene has a single allelic mutation namely, the gdhB-227. It was determined that MDR isolate carried bap, basABCDFGHIJ, csuA/BABCDE, bauABCDEF, plcD, pgaABCD, entE, barAB, ompA, abaIR, piT2EAFTE/AUBl, fimADT, cvaC, bfmR, bfmS virulence genes. In our study imipenem induced the highest biofilm formation at a concentration of 32 µg/ml and levofloxacin at a concentration of 16 µg/ml. In conclusion, we detected a new MDR A. baumannii ST2121 clone harboring blaOXA-66 gene that has been reported for the first time in Turkey.

Author(s):  
Niloofar Tafreshi ◽  
Laleh Babaeekhou ◽  
Maryam Ghane

Background and Objectives: Notwithstanding the increased prevalence of Acinetobacter baumannii drug-resistant isolates, treatment options are progressively limiting. This study aims to provide a recent report on antibiotic susceptibility in burn wound isolates of A. baumannii, and the importance of OXA beta-lactamases in carbapenem resistance. Materials and Methods: The susceptibility levels to different antimicrobial categories were determined among 84 A. baumannii isolates from burn wound infection between 2016 and 2018. Multiplex PCR was used to detect OXA beta-lactamases genes, including blaOXA-51, blaOXA-23, blaOXA-24 and blaOXA-58. ISAba-1 association with blaOXA-51, blaOXA-23 and blaOXA-58 was detected by PCR mapping. Results: All the isolates were determined as multidrug-resistant (MDR) and 69% as extensively drug-resistant (XDR). Different carbapenems MIC ranges (MIC50 and MIC90) were observed among the isolates harboring blaOXA-like genes and isolates with the OXA-24-like enzyme showed higher carbapenems MIC ranges. The prevalence of blaOXA-51-like, blaOXA-23-like, blaOXA-24-like and blaOXA-58-like were 100%, 53.57%, 41.66% and 30.95%, respectively. ISAba-1 insertion sequence was found to be upstream to blaOXA-23-like and blaOXA-58-like genes in 23 out of 45 (71.1%) blaOXA-23-like-positive and 4 out of 23 (15.3) blaOXA-58-like-positive isolates, respectively. Conclusion: Resistance to carbapenems as the last resort for treatment of A. baumannii infections is growing. This study, for the first time in Iran, has observed the increased frequency of blaOXA-24-like and blaOXA-58-like genes and found an association between ISAba-1 and blaOXA-58-like gene, which signifies the possible risk of increased diversity in OXA beta-lactamases and growth in carbapenem resistance


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Udomluk Leungtongkam ◽  
Rapee Thummeepak ◽  
Thawatchai Kitti ◽  
Kannipa Tasanapak ◽  
Jintana Wongwigkarn ◽  
...  

Abstract In this study, we examined the association between antimicrobial resistance, CRISPR/Cas systems and virulence with phage susceptibility in Acinetobacter baumannii and investigated draft genomes of phage susceptible multidrug resistant A. baumannii strains from Thailand. We investigated 230 A. baumannii strains using 17 lytic A. baumannii phages and the phage susceptibility was 46.5% (107/230). Phage susceptibility was also associated with resistance to numerous antibiotics (p-value < 0.05). We also found association between biofilm formation and the presence of ompA gene among phage susceptible A. baumannii strains (p-value < 0.05). A. baumannii isolates carrying cas5 or combinations of two or three other cas genes, showed a significant increase in phage resistance. Whole-genome sequences of seven phage susceptible A. baumannii isolates revealed that six groups of antibiotic resistance genes were carried by all seven phage susceptible A. baumannii. All strains carried biofilm associated genes and two strains harbored complete prophages, acquired copper tolerance genes, and CRISPR-associated (cas) genes. In conclusion, our data exhibits an association between virulence determinants and biofilm formation among phage susceptible A. baumannii strains. These data help to understand the bacterial co-evolution with phages.


Biofouling ◽  
2020 ◽  
Vol 36 (4) ◽  
pp. 492-504 ◽  
Author(s):  
Saima Muzammil ◽  
Mohsin Khurshid ◽  
Iqra Nawaz ◽  
Muhammad Hussnain Siddique ◽  
Muhammad Zubair ◽  
...  

Author(s):  
María-Guadalupe Avila-Novoa ◽  
Oscar-Alberto Solís-Velázquez ◽  
Daniel-Eduardo Rangel-López ◽  
Jean-Pierre González-Gómez ◽  
Pedro-Javier Guerrero-Medina ◽  
...  

Acinetobacter baumannii is an important opportunistic pathogen that shows resistance to cephalosporins, penicillins, carbapenems, fluoroquinolones, and aminoglycosides, the multiresistance being associated with its ability to form biofilms in clinical environments. The aim of this study was to determine biofilm formation and its potential association with genes involved in antibiotic resistance mechanisms of A. baumannii isolates of different clinical specimens. We demonstrated 100% of the A. baumannii isolates examined to be multidrug resistant (MDR), presenting a 73.3% susceptibility to cefepime and a 53.3% susceptibility to ciprofloxacin. All A. baumannii isolates were positive for blaOXA-51, 33.3% being positive for blaOXA-23 and ISAba1, and 73.3% being positive for gyrA. We found 86.6% of A. baumannii strains to be low-grade biofilm formers and 13.3% to be biofilm negative; culturing on Congo red agar (CRA) plates revealed that 73.3% of the A. baumannii isolates to be biofilm producers, while 26.6% were not. These properties, combined with the role of A. baumannii as a nosocomial pathogen, increase the probability of A. baumannii causing nosocomial infections and outbreaks as a complication during therapeutic treatments and emphasize the need to control A. baumannii biofilms in hospital environments.


Antibiotics ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1115
Author(s):  
Hui-Ling Lin ◽  
Chen-En Chiang ◽  
Mei-Chun Lin ◽  
Mei-Lan Kau ◽  
Yun-Tzu Lin ◽  
...  

Limited therapeutic options are available for multidrug-resistant Acinetobacter baumannii (MDR-AB), and the development of effective treatments is urgently needed. The efficacy of four aerosolized antibiotics (gentamicin, amikacin, imipenem, and meropenem) on three different MDR-AB strains was evaluated using hypertonic saline (HS, 7 g/100 mL) as the aerosol carrier. HS aerosol effectively hindered biofilm formation by specific MDR-AB strains. It could also interrupt the swarming dynamics of MDR-AB and the production of extracellular polymeric substances, which are essential for biofilm progression. Biofilms protect the microorganisms from antibiotics. The use of HS aerosol as a carrier resulted in a decreased tolerance to gentamicin and amikacin in the biofilm-rich MDR-AB. Moreover, we tested the aerosol characteristics of antibiotics mixed with HS and saline, and results showed that HS enhanced the inhaled delivery dose with a smaller particle size distribution of the four antibiotics. Our findings demonstrate the potential of using “old” antibiotics with our “new” aerosol carrier, and potentiate an alternative therapeutic strategy to eliminate MDR-AB infections from a biofilm-disruption perspective.


2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Yamuna Chand ◽  
Sujan Khadka ◽  
Sanjeep Sapkota ◽  
Suprina Sharma ◽  
Santosh Khanal ◽  
...  

The multidrug- or extensively drug-resistant (MDR/XDR) Pseudomonas aeruginosa carrying some virulence genes has become a global public health threat. However, in Nepal, there is no existing report showing the prevalence of oprL and toxA virulence genes among the clinical isolates of P. aeruginosa. Therefore, this study was conducted for the first time in the country to detect the virulence genes (oprL and toxA) and antibiotic susceptibility pattern of P. aeruginosa. A total of 7,898 clinical specimens were investigated following the standard microbiological procedures. The antibiotic susceptibility testing was examined by the modified disc diffusion method, and virulence genes oprL and toxA of P. aeruginosa were assessed using multiplex PCR. Among the analyzed specimens, 87 isolates were identified to be P. aeruginosa of which 38 (43.68%) isolates were reported as MDR. A higher ratio of P. aeruginosa was detected from urine samples 40 (45.98%), outpatients’ specimens 63 (72.4%), and in patients of the age group of 60–79 years 36 (41.37%). P. aeruginosa was more prevalent in males 56 (64.36%) than in female patients 31 (35.63%). Polymyxin (83.90%) was the most effective antibiotic. P. aeruginosa (100%) isolates harboured the oprL gene, while 95.4% of isolates were positive for the toxA gene. Identification of virulence genes such as oprL and toxA carrying isolates along with the multidrug resistance warrants the need for strategic interventions to prevent the emergence and spread of antimicrobial resistance (AMR). The findings could assist in increasing awareness about antibiotic resistance and suggest the judicious prescription of antibiotics to treat the patients in clinical settings of Nepal.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
K. Bierowiec ◽  
M. Miszczak ◽  
A. Korzeniowska-Kowal ◽  
A. Wzorek ◽  
D. Płókarz ◽  
...  

AbstractStaphylococcus pseudintermedius is a well-known coagulase-positive staphylococcus that is mainly associated with the asymptomatic colonization of the skin of pets and mucous membranes. Little is still known about the occurrence of S. pseudintermedius in cats. The current study aimed to characterize the isolates of S. pseudintermedius from sick and healthy cats. This was achieved by examining their antibiotic resistance properties, biofilm formation, and genotype differences. Six hundred and seventy-six cats were swabbed (595 healthy and 81 sick cats). Thirty-five distinct S. pseudintermedius isolates from 27 cats were isolated. The prevalence of S. pseudintermedius in healthy and sick cats was 2.49% and 7.61%, respectively. In comparison, MRSP (methicillin-resistant Staphylococcus pseudintermedius) prevalence was 0.12% and 2.98%, respectively. Cats were more frequently colonized with S. pseudintermedius when kept with dogs, regardless of their health condition, with this result being statistically significant. Multidrug resistance was detected in 50%, and 38.46% of S. pseudintermedius isolates from healthy and sick cats, respectively. In contrast, genetic multidrug resistance was detected in 59% and 46.15% cases, respectively. Seven from eight isolated MRSPs were multidrug-resistant. Multi-locus sequence typing (MLST) assigned isolates to 19 types, of which 16 types submitted for the first time to the PubMLST database. The most frequently detected STs (sequence types) were 551 and 71. ST71 and ST551 were mainly isolated from cats with clinical signs of infection. All were MRSPs, regardless of cats’ health. These isolates were characterized with the most frequent antibiotic resistance at the phenotypic and genotypic level.


Pathogens ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 630 ◽  
Author(s):  
Aisha M. Alamri ◽  
Afnan A. Alsultan ◽  
Mohammad A. Ansari ◽  
Amani M. Alnimr

This study analyzed the genotype, antibiotic resistance, and biofilm formation of Acinetobacter baumannii strains and assessed the correlation between biofilm formation, antibiotic resistance, and biofilm-related risk factors. A total of 207 non-replicate multi-drug-resistant A. baumannii strains were prospectively isolated. Phenotypic identification and antimicrobial susceptibility testing were carried out. Isolate biofilm formation ability was evaluated using the tissue culture plate (TCP), Congo red agar, and tube methods. Clonal relatedness between the strains was assessed by enterobacterial repetitive intergenic consensus-PCR genotyping. Of the 207 isolates, 52.5% originated from an intensive care unit setting, and pan resistance was observed against ceftazidime and cefepime, with elevated resistance (99–94%) to piperacillin/tazobactam, imipenem, levofloxacin, and ciprofloxacin. alongside high susceptibility to tigecycline (97.8%). The Tissue culture plate, Tube method, and Congo red agar methods revealed that 53.6%, 20.8%, and 2.7% of the strains were strong biofilm producers, respectively, while a significant correlation was observed between biofilm formation and device-originating respiratory isolates (p = 0.0009) and between biofilm formation in colonized vs. true infection isolates (p = 0.0001). No correlation was detected between antibiotic resistance and biofilm formation capacity, and the majority of isolates were clonally unrelated. These findings highlight the urgent need for implementing strict infection control measures in clinical settings.


2013 ◽  
Vol 79 (21) ◽  
pp. 6535-6543 ◽  
Author(s):  
H. M. Sharon Goh ◽  
Scott A. Beatson ◽  
Makrina Totsika ◽  
Danilo G. Moriel ◽  
Minh-Duy Phan ◽  
...  

ABSTRACTAcinetobacter baumanniiis a multidrug-resistant pathogen associated with hospital outbreaks of infection across the globe, particularly in the intensive care unit. The ability ofA. baumanniito survive in the hospital environment for long periods is linked to antibiotic resistance and its capacity to form biofilms. Here we studied the prevalence, expression, and function of theA. baumanniibiofilm-associated protein (Bap) in 24 carbapenem-resistantA. baumanniiST92 strains isolated from a single institution over a 10-year period. Thebapgene was highly prevalent, with 22/24 strains being positive forbapby PCR. Partial sequencing ofbapwas performed on the index case strain MS1968 and revealed it to be a large and highly repetitive gene approximately 16 kb in size. Phylogenetic analysis employing a 1,948-amino-acid region corresponding to the C terminus of Bap showed that BapMS1968clusters with Bap sequences from clonal complex 2 (CC2) strains ACICU, TCDC-AB0715, and 1656-2 and is distinct from Bap in CC1 strains. By using overlapping PCR, thebapMS1968gene was cloned, and its expression in a recombinantEscherichia colistrain resulted in increased biofilm formation. A Bap-specific antibody was generated, and Western blot analysis showed that the majority ofA. baumanniistrains expressed an ∼200-kDa Bap protein. Further analysis of three Bap-positiveA. baumanniistrains demonstrated that Bap is expressed at the cell surface and is associated with biofilm formation. Finally, biofilm formation by these Bap-positive strains could be inhibited by affinity-purified Bap antibodies, demonstrating the direct contribution of Bap to biofilm growth byA. baumanniiclinical isolates.


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