scholarly journals Phenotypic and Genotypic Evaluation of Antibiotic Resistance of Acinetobacter baumannii Bacteria Isolated from Surgical ICU Patients in Pakistan

2021 ◽  
Vol 14 (4) ◽  
Author(s):  
Nureen Zahra ◽  
Basit Zeshan ◽  
Muhammad Mubeen Ali Qadri ◽  
Musarat Ishaq ◽  
Muhammad Afzal ◽  
...  

Background: Carbapenem-resistant Acinetobacter baumannii (CRAB) is a significant nosocomial pathogen, causing serious threats concerning community-wide outbreaks globally, as well as in Pakistan. Antimicrobial resistance in A. baumannii is increasing day by day. Objectives: The study aimed to find out the antibiotic resistance (AMR) patterns and evaluate the AMR genes in clinical isolates from patients admitted to the surgical Intensive Care units (ICUs) at different hospitals in Lahore, Pakistan. Methods: A total of 593 clinical specimens were collected from patients admitted to the surgical ICUs of three different local hospitals in Lahore, Pakistan. From these samples, a total of 90 A. baumannii isolates were identified and further investigated to observe phenotypic resistance patterns and detect carbapenemases resistance genes. Results: The results showed that phenotypic resistance against amikacin was 27.2%, ceftriaxone 100%, ceftazidime 27.2%, cefepime 63.3%, ciprofloxacin and co-trimoxazole 100%, gentamicin 40%, imipenem 22.2%, meropenem 21.1%, piperacillin-tazobactam 27.2%, tigecycline 27.2%, and tetracycline 63.3%. All A. baumannii isolates were found to be sensitive to colistin (CT), polymixin-B (PB), and tobramycin (TOB). The PCR amplification of carbapenemases genes revealed the prevalence of blaOXA-23, blaOXA-51, and blaOXA-40 in 73, 90, and 64.4% of the isolates, respectively, along with blaNDM1 (92.2%), blaVIM (40%), blaIMP (90%), ISAba1 (85.5%), sul1 (16.6%), sul2 (20%), armA (32.2%), and PER-1 (12%) while the blaOXA-24 and blaOXA-58 genes were not detected in the isolates. The sequence analysis of the blaOXA-23 and blaOXA-51 genes showed 98% and 95% similarity with previously reported sequences in the GenBank database. Conclusions: The present study indicated that the emergence of high carbapenem resistance in CRAB isolates has increased, which may pose serious limitations in the choice of drugs for nosocomial infections.

Author(s):  
Rabia Arshad

Background: Antimicrobial resistance is one of the research priorities of health organizations due to increased risk of morbidity and mortality. Outbreaks of nosocomial infections caused by carbapenem-resistant Acinetobacter Baumannii (CRAB) strains are at rise worldwide. Antimicrobial resistance to carbapenems reduces clinical therapeutic choices and frequently led to treatment failure. The aim of our study was to determine the prevalence of carbapenem resistance in A. baumannii isolated from patients in intensive care units (ICUs). Methods: This cross-sectional study was carried out in the Department of Microbiology, Basic Medical Sciences Institute (BMSI), Jinnah Postgraduate Medical Centre (JPMC), Karachi, from December 2016 to November 2017. Total 63 non-repetitive A. baumannii were collected from the patients’ specimens, admitted to medical and surgical ICUs and wards of JPMC, Karachi. The bacterial isolates were processed according to standard microbiological procedures to observe for carbapenem resistance. SPSS 21 was used for data analysis. Results: Out of the 63 patients, 40 (63.5%) were male. The age of the patient ranged from 15-85 year, with average of 43 year. 34.9% patients had been hospitalized for 3 days. Chronic obstructive pulmonary disease was present in highest number with average of 58.7% for morbidity. Number of patients on mechanical ventilation was highest (65.1%). All isolates were susceptible to colistin. The resistance to ampicillin-sulbactam, ceftazidime, ciprofloxacin, amikacin, piperacillin- tazobactam and meropenem was 82.5%, 81%, 100%, 87.3%, 82.5% and 82% respectively. Out of 82% CRAB, 77% were obtained from ICUs. Conclusion: This study has revealed the high rate of carbapenem resistance in A. baumannii isolates in ICUs thus leaving behind limited therapeutic options.


Antibiotics ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1054
Author(s):  
Nalumon Thadtapong ◽  
Soraya Chaturongakul ◽  
Sunhapas Soodvilai ◽  
Padungsri Dubbs

Resistance to the last-line antibiotics against invasive Gram-negative bacterial infection is a rising concern in public health. Multidrug resistant (MDR) Acinetobacter baumannii Aci46 can resist colistin and carbapenems with a minimum inhibitory concentration of 512 µg/mL as determined by microdilution method and shows no zone of inhibition by disk diffusion method. These phenotypic characteristics prompted us to further investigate the genotypic characteristics of Aci46. Next generation sequencing was applied in this study to obtain whole genome data. We determined that Aci46 belongs to Pasture ST2 and is phylogenetically clustered with international clone (IC) II as the predominant strain in Thailand. Interestingly, Aci46 is identical to Oxford ST1962 that previously has never been isolated in Thailand. Two plasmids were identified (pAci46a and pAci46b), neither of which harbors any antibiotic resistance genes but pAci46a carries a conjugational system (type 4 secretion system or T4SS). Comparative genomics with other polymyxin and carbapenem-resistant A. baumannii strains (AC30 and R14) identified shared features such as CzcCBA, encoding a cobalt/zinc/cadmium efflux RND transporter, as well as a drug transporter with a possible role in colistin and/or carbapenem resistance in A. baumannii. Single nucleotide polymorphism (SNP) analyses against MDR ACICU strain showed three novel mutations i.e., Glu229Asp, Pro200Leu, and Ala138Thr, in the polymyxin resistance component, PmrB. Overall, this study focused on Aci46 whole genome data analysis, its correlation with antibiotic resistance phenotypes, and the presence of potential virulence associated factors.


2016 ◽  
Vol 54 (7) ◽  
pp. 1700-1710 ◽  
Author(s):  
Thomas J. Gniadek ◽  
Karen C. Carroll ◽  
Patricia J. Simner

The non-glucose-fermenting Gram-negative bacilliPseudomonas aeruginosaandAcinetobacter baumanniiare increasingly acquiring carbapenem resistance. Given their intrinsic antibiotic resistance, this can cause extremely difficult-to-treat infections. Additionally, resistance gene transfer can occur between Gram-negative species, regardless of their ability to ferment glucose. Thus, the acquisition of carbapenemase genes by these organisms increases the risk of carbapenemase spread in general. Ultimately, infection control practitioners and clinical microbiologists need to work together to determine the risk carried by carbapenem-resistant non-glucose-fermenting Gram-negative bacilli (CR-NF) in their institution and what methods should be considered for surveillance and detection of CR-NF.


2019 ◽  
Vol 70 (10) ◽  
pp. 3502-3507
Author(s):  
Irina Gheorghe ◽  
Violeta Corina Cristea ◽  
Luminita Marutescu ◽  
Marcela Popa ◽  
Carmen Murariu ◽  
...  

We aimed to identify the virulence and antimicrobial resistance features in Carbapenem Resistant Acinetobacter baumannii (CRAB) strains isolated from hospital settings and compare them with those isolated in the same period of time from community acquired (CA) infections in Bucharest, south of Romania. A total number of 93 A. baumannii strains were isolated in majority from hospitalized patients and from CA infections. The resistance and virulence mechanisms of the strains were characterized by phenotypic and genotypic methods. The antibiotic resistance profiles in H and CA A. baumannii isolates revealed high percentages of carbapenem-resistance in both H and CA isolates. The ciprofloxacin resistance was found very closed in both types of isolates (84%/83.33%). CRAB H and CA isolates revealed the intrinsec carbapenemase OXA-51and the acquired carbapenemases OXA-23, OXA-24, IMP,and VIM-2. The blaOXA-23 gene was identified in different plasmid types (GR2-Aci1, GR6-pACICU2). rep135040, p3S18 and Aci6 in H A. baumannii isolates. The most frequently expressed virulence factor was lipase and DN-ase. OXA-51-like alleles corresponding to the two main sequence groups were identified as blaOXA66 (63.63% of the isolates) and respectively, blaOXA-69 (38.39%) and revealed the corresponding type of ompAand csuE sequence grouping. AphA6 (24%/16.6%), AphA1 (16%/16.6%) and aadB (9.3%/5.5%) genes were responsible for aminoglycosides resistance. Our survey revealed a high drug resistance in A. baumannii isolates. Different plasmid groups containing CRAB isolates may facilitate the blaOXA23 dissemination.


2021 ◽  
Vol 11 (4) ◽  
pp. 642-657
Author(s):  
Shimaa Tawfeeq Omara ◽  
Ashraf Samir Hakim ◽  
Magdy Ali Bakry

Detailed information on the resistance patterns of Staphylococcus aureus (S. aureus) in milk and cheese is strongly required to facilitate risk assessment analysis in case of food poisoning context and to improve therapeutic approaches used in dairy farms. The present study aimed to perform phenotypic and genotypic antimicrobial characterizations of methicillin, vancomycin, and erythromycin-resistant S. aureus isolated from milk and dairy products through screening mecA, vanA, and ermC using molecular PCR amplification technology. Moreover, the association between each genotypic and its related antibiotic resistance phenotypic features within the isolated S. aureus strains were analyzed. Moreover, the current study aimed to study MRSA's ability to form biofilms. Out of 226 milk and dairy product samples collected from different retailers in Giza Governorate, 69.5% of the samples were positive for the presence of S. aureus. The isolation rate of S. aureus strains from cattle milk, sheep milk, white cheese, flamenco, and mesh samples were 79.7%, 76.5%, 56.0%, 40.0%, and 94.7%, respectively. Multidrug-resistant S. aureus (MDR) was detected in 51% of all isolated S. aureus strains. All tested S. aureus strains were sensitive to trimethoprim-sulfamethoxazole, linezolid, ciprofloxacin, and gentamycin. However, their resistance rates against penicillin, oxacillin, vancomycin, erythromycin, tetracycline, clindamycin and chloramphenicol were 62.4%, 65.0%, 44.6%, 45.9%, 21.0%, 14.0%, and 2.5%, respectively. Of the isolated S. aureus strains, 72.6%, 40.1%, and 48.4% were carriers for mecA, vanA, and ermC genes and the amplified products were at 310, 1030, and 295 bp, respectively. Methicillin-resistant S. aureus isolates were detected in 47.1% of all isolated S. aureus strains. The results indicated that 35.0% of the tested S. aureus strains were genotypic vanA gene carriers and phenotypic resistant to vancomycin representing vancomycin-resistant S. aureus strains. Moreover, 42.7% of all isolated S. aureus strains were carriers for ermC gene and were phenotypic resistant to erythromycin representing erythromycin-resistant S. aureus. The presence of mecA, vanA, and ermC genes in S. aureus was statistically associated with their related phenotypic resistance patterns against both penicillin and oxacillin, vancomycin, and erythromycin, respectively. Moreover, along with an increase in the frequency of mecA, vanA, and ermC genes, their phenotypic antibiotic resistance patterns sharply increased with an odd ratio >1. Of MRSA isolates, 6.8% indicated weak biofilm-formation ability, while 93.2% exhibit no biofilm-forming ability.


Author(s):  
Alaa Abouelfetouh ◽  
Aisha S. Torky ◽  
Elsayed Aboulmagd

Abstract Background Antibiotic use is largely under-regulated in Egypt leading to the emergence of resistant isolates. Carbapenems are last resort agents to treat Acinetobacter baumannii infections resistant to other classes of antibiotics. However, carbapenem-resistant isolates are emerging at an alarming rate. This study aimed at phenotypically and molecularly characterizing seventy four carbapenem-unsusceptible A. baumannii isolates from Egypt to detect the different enzymes responsible for carbapenem resistance. Methods Carbapenemase production was assessed by a number of phenotypic methods: modified Hodge test (MHT), carbapenem inactivation method (CIM), combined disc test (CDT), CarbAcineto NP test and boronic acid disc test. Polymerase chain reaction (PCR) was used to screen the isolates for the presence of some genes responsible for resistance to carbapenems, as well as some insertion sequences. Results PCR amplification of class D carbapenemases revealed the prevalence of blaOXA-51 and blaOXA-23 in 100% of the isolates and of blaOXA-58 in only one isolate (1.4%). blaVIM and blaNDM-1 belonging to class B metallo-β-lactamases were present in 100 and 12.1% of the isolates, respectively. The prevalence of ISAba1, ISAba2 and ISAba3 was 100, 2.7 and 4.1%, respectively. None of the tested isolates carried blaOXA-40, blaIMP, blaSIM, blaSPM, blaGIM or the class A blaKPC. Taking PCR as the gold standard method for the detection of different carbapenemases, the sensitivities of the MHT, CIM, CDT, CarbAcineto NP test and boronic acid disc/imipenem or meropenem test for this particular collection of isolates were 78.4, 68.9, 79.7, 95.9, and 56.8% or 70.3%, respectively. Conclusions The widespread detection of carbapenem-resistant A. baumannii (CR-AB) has become a real threat to the efficacy of treatment regimens. Among the studied cohort of CR-AB clinical isolates, blaOXA-51, blaOXA-23 and blaVIM were the most prevalent, followed by blaNDM-1 and blaOXA-58. The genotypic detection of carbapenemases among CR-AB clinical isolates using PCR was most conclusive, followed closely by the phenotypic testing using CarbAcineto NP test.


2019 ◽  
Vol 70 (10) ◽  
pp. 3502-3507

We aimed to identify the virulence and antimicrobial resistance features in Carbapenem Resistant Acinetobacter baumannii (CRAB) strains isolated from hospital settings and compare them with those isolated in the same period of time from community acquired (CA) infections in Bucharest, south of Romania. A total number of 93 A. baumannii strains were isolated in majority from hospitalized patients and from CA infections. The resistance and virulence mechanisms of the strains were characterized by phenotypic and genotypic methods. The antibiotic resistance profiles in H and CA A. baumannii isolates revealed high percentages of carbapenem-resistance in both H and CA isolates. The ciprofloxacin resistance was found very closed in both types of isolates (84%/83.33%). CRAB H and CA isolates revealed the intrinsec carbapenemase OXA-51and the acquired carbapenemases OXA-23, OXA-24, IMP,and VIM-2. The blaOXA-23 gene was identified in different plasmid types (GR2-Aci1, GR6-pACICU2). rep135040, p3S18 and Aci6 in H A. baumannii isolates. The most frequently expressed virulence factor was lipase and DN-ase. OXA-51-like alleles corresponding to the two main sequence groups were identified as blaOXA66 (63.63% of the isolates) and respectively, blaOXA-69 (38.39%) and revealed the corresponding type of ompAand csuE sequence grouping. AphA6 (24%/16.6%), AphA1 (16%/16.6%) and aadB (9.3%/5.5%) genes were responsible for aminoglycosides resistance. Our survey revealed a high drug resistance in A. baumannii isolates. Different plasmid groups containing CRAB isolates may facilitate the blaOXA23 dissemination. Keywords: carbapenem resistance, virulence, community acquired, nosocomial infections


2021 ◽  
Vol 71 (11) ◽  
pp. 2576-2581
Author(s):  
Saima Ishtiaq ◽  
Sidrah Saleem ◽  
Abdul Waheed ◽  
Arslan Ahmed Alvi

Objective: To evaluate carbapenem resistance and to detect blaOXA-23 and blaOXA-51 genes in carbapenem-resistant acinetobacter baumanii isolates recovered from patients having pneumonia secondry to ventilation. Methods: The cross-sectional study was conducted from July 2017 to June 2018 at the Department of Microbiology, University of Health Sciences, Lahore, Pakistan, and comprised endotracheal aspirates / tracheobroncheal lavage samples from patients irrespective of age and gender who developed pneumonia after being on the ventilator for 48 hrs at the Combined Military Hospital, and Jinnah Hospital, Lahore.  The samples were inoculated on MacConkey and blood agar and aerobically incubated at a temperature of 370C for 18-24 hours. The isolated organisms were further assessed by standard morphological, cultural and biochemical profile. Antibiotic susceptibility was done by Kirby-Bauer disc diffusion method. Carbapenem-resistant acinetobacter baumannii were checked for carbapenemase production using Modified Hodge Test. Conventional polymerase chain reaction and agarose gel electrophoreses were performed to detect blaOXA-23 and blaOXA-51 genes. Data was analysed using SPSS 17. Results: Out of 157 samples, 92(58.6%) yielded growth of bacteria, and, among them, 39(42.4%) were identified as acinetobacter baumannii. All (100%) acinetobacter baumannii cases showed resistance to carbapenem, were producing carbapenemase enzyme, and were positive for blaOXA-51 gene. The blaOXA-23 gene was amplified in 38(97.4%) isolates. Conclusion: BlaOXA-23 gene appeared to be the major cause of carbapenem resistance. Continuous...


2020 ◽  
Author(s):  
Reem M Hassan ◽  
Sherifa T Salem ◽  
Saly Ismail Mostafa Hassan ◽  
Asmaa Sayed Hegab ◽  
Yasmine S Elkholy

AbstractAcinetobacter baumannii (A. baumannii) represents a global threat owing to its ability to resist most of the currently available antimicrobial agents. Moreover, emergence of carbapenem resistant A. baumannii (CR-AB) isolates limits the available treatment options. Enzymatic degradation by variety of ß-lactamases, have been identified as the most common mechanism of carbapenem resistance in A. baumannii. The alarming increase in the prevalence of CR-AB necessitates continuous screening and molecular characterization to appreciate the problem. The present study was performed to assess the prevalence and characterize carbapenemases among 206 CR-AB isolated from various clinical specimens collected from different intensive care units at Kasr Al-Aini Hospital.All isolates were confirmed to be A. baumannii by detection of the blaOXA-51-like gene. Molecular screening of 13 common Ambler class bla carbapenemases genes in addition to insertion sequence (IS-1) upstream OXA-23 was performed by using four sets of multiplex PCR, followed by identification using gene sequencing technology. Among the investigated genes, the prevalence of blaOXA-23, and blaOXA-58 were 77.7%, and 1.9%, respectively. The ISAba1 was detected in 10% of the blaOXA-23 positive isolates. The prevalence of metallo-β-lactamases (MBLs) studied; blaNDM-1, blaSPM, blaVIM, blaSIM-1 were 11.7%, 6.3%, 0.5%, and 0.5% respectively. One of class A; bla KPC was detected in 10.7% of the investigated isolates. blaOXA-24/40, blaIMP, blaGES, blaVEB and blaGIM were not detected in any of the studied isolates. Moreover, 18.4% of the isolates have shown to harbor two or more of the screened bla genes. We concluded that the most prevalent type of ß-lactamases genes among CR-AB isolates collected from Egyptian patients were blaOXA-23 followed by blaNDM-1 and blaKPC.Author summaryCarbapenem-resistant A. baumannii has become a real global health threat. The aim of the present study was to characterize and to assess the prevalence of carbapenemases among 206 CR-AB clinical isolates from Egyptian patients. We concluded that the most prevalent type of ß-lactamases genes among CR-AB isolates collected from Egyptian patients were blaOXA-23 followed by blaNDM-1 and blaKPC. In this study, ISAba1 was detected upstream 10% of blaOXA-23 positive isolates only which indicates that the spread of resistance among Acinetobacter isolates could be either chromosomal or plamid-mediated.


2021 ◽  
Vol 8 ◽  
Author(s):  
Amit Sharma ◽  
Rajni Gaind

Background:Acinetobacter calcoaceticus–baumannii (ACB) complex has emerged as an important nosocomial pathogen and is associated with life-threatening infections, especially among ICU patients, including neonates. Carbapenem resistance in Acinetobacter baumannii has emerged globally and is commonly mediated by blaOXA-23. Clinically significant infections with carbapenem-resistant Acinetobacter baumannii (CRAB) are a major concern since therapeutic options are limited and associated mortality is high. Early diagnosis of both the pathogen and resistance is important to initiate the optimal therapy and prevent selection of resistance. In the current study, a loop-mediated isothermal amplification (LAMP) assay was developed for rapid detection of the ACB complex and carbapenem resistance mediated by blaOXA-23.Methodology: Universal LAMP primers were designed for the detection of significant members of the ACB complex and carbapenem resistance targeting the ITS 16S–23S rRNA and blaOXA-23 gene respectively. The optimal conditions for the LAMP assay were standardized for each primer set using standard ATCC strains. The sensitivity of the LAMP assay was assessed based on the limit of detection (LOD) using different DNA concentrations and colony counts. The specificity of LAMP was determined using the non-ACB complex and non-Acinetobacter species. The results of the LAMP assay were compared with those of polymerase chain reaction (PCR).Results: The optimal temperature for the LAMP assay was 65°C, and the detection time varied with various primers designed. Using the ITS Ab1 primer, LODs of LAMP and PCR assays were 100 pg/μl and 1 ng/μl of DNA concentration and 104 cfu/ml and 108 cfu/ml of colony count, respectively. The LAMP assay was 10- and 104-fold more sensitive than PCR using DNA concentration and colony count, respectively. The LAMP assay was found to be specific for clinically important ACB complex species.Significance of the study: The LAMP assay can be applied for early detection of significant species of the ACB complex from clinical samples and their carbapenem-resistant variants. Depending on the emerging pathogen and locally prevalent resistance genes, the LAMP assay can be modified for detection of colonization or infection by various resistant bugs.


Sign in / Sign up

Export Citation Format

Share Document