scholarly journals Molecular and phenotypic characterization of efflux pump and biofilm in multi-drug resistant non-typhoidal Salmonella Serovars isolated from food animals and handlers in Lagos Nigeria

2021 ◽  
Vol 3 (1) ◽  
Author(s):  
Elizabeth Tolulope Olubisose ◽  
Abraham Ajayi ◽  
Adeyemi Isaac Adeleye ◽  
Stella Ifeanyi Smith
Keyword(s):  
Antibiotic Resistance ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Phenotypic Characterization ◽  
Pump Activity ◽  
Food Animals

Abstract Background Multidrug resistance efflux pumps and biofilm formation are mechanisms by which bacteria can evade the actions of many antimicrobials. Antibiotic resistant non-typhoidal Salmonella serovars have become wide spread causing infections that result in high morbidity and mortality globally. The aim of this study was to evaluate the efflux pump activity and biofilm forming capability of multidrug resistant non-typhoidal Salmonella (NTS) serovars isolated from food handlers and animals (cattle, chicken and sheep) in Lagos. Methods Forty eight NTS serovars were subjected to antibiotic susceptibility testing by the disc diffusion method and phenotypic characterization of biofilm formation was done by tissue culture plate method. Phenotypic evaluation of efflux pump activity was done by the ethidium bromide cartwheel method and genes encoding biofilm formation and efflux pump activity were determined by PCR. Results All 48 Salmonella isolates displayed resistance to one or more classes of test antibiotics with 100% resistance to amoxicillin-clavulanic acid. Phenotypically, 28 (58.3%) of the isolates exhibited efflux pump activity. However, genotypically, 7 (14.6%) of the isolates harboured acrA, acrB and tolC, 8 (16.7%) harboured acrA, acrD and tolC while 33 (68.8%) possessed acrA, acrB, acrD and tolC. All (100%) the isolates phenotypically had the ability to form biofilm with 23 (47.9%), 24 (50.0%), 1 (2.1%) categorized as strong (SBF), moderate (MBF) and weak (WBF) biofilm formers respectively but csgA gene was detected in only 23 (47.9%) of them. Antibiotic resistance frequency was significant (p < 0.05) in SBF and MBF and efflux pump activity was detected in 6, 21, and 1 SBF, MBF and WBF respectively. Conclusion These data suggest that Salmonella serovars isolated from different food animals and humans possess active efflux pumps and biofilm forming potential which has an interplay in antibiotic resistance. There is need for prudent use of antibiotics in veterinary medicine and scrupulous hygiene practice to prevent the transmission of multidrug resistant Salmonella species within the food chain.

scholarly journals Evaluation of efflux pump activity and biofilm formation in multidrug resistant clinical isolates of Pseudomonas aeruginosa isolated from a Federal Medical Center in Nigeria

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Florence Chijindu Ugwuanyi ◽  
Abraham Ajayi ◽  
David Ajiboye Ojo ◽  
Adeyemi Isaac Adeleye ◽  
Stella Ifeanyi Smith
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Medical Center ◽  
Opportunistic Pathogen ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Biochemical Tests ◽  
Pump Activity

Abstract Background Pseudomonas aeruginosa an opportunistic pathogen, is widely associated with nosocomial infections and exhibits resistance to multiple classes of antibiotics. The aim of this study was to determine the antibiotic resistance profile, biofilm formation and efflux pump activity of Pseudomonas strains isolated from clinical samples in Abeokuta Ogun state Nigeria. Methods Fifty suspected Pseudomonas isolates were characterized by standard biochemical tests and PCR using Pseudomonas species -specific primers. Antibiotic susceptibility testing was done by the disc diffusion method. Efflux pump activity screening was done by the ethidium bromide method and biofilm formation assay by the tissue plate method. Genes encoding biofilm formation (pslA & plsD) and efflux pump activity (mexA, mexB and oprM) were assayed by PCR. Results Thirty-nine Pseudomonas spp. were identified of which 35 were Pseudomonas aeruginosa and 4 Pseudomonas spp. All 39 (100%) Pseudomonas isolates were resistant to ceftazidime, cefuroxime and amoxicillin-clavulanate. Thirty-six (92%), 10(25.6%), 20 (51.2%), 11(28%) and 9(23%) of the isolates were resistant to nitrofurantoin, imipenem, gentamicin, cefepime and aztreonam respectively. All the isolates had the ability to form biofilm and 11 (28%) of them were strong biofilm formers. They all (100%) harboured the pslA and pslD biofilm encoding genes. Varied relationships between biofilm formation and resistance to ciprofloxacin, ofloxacin, cefixime, gentamicin, imipenem, and aztreonam were observed. Only 23(59%) of the Pseudomonas isolates phenotypically exhibited efflux pump activity but mexA gene was detected in all 39 (100%) isolates while mexB and oprM genes were detected in 91%, 92%, and 88% of strong, moderate and weak biofilm formers respectively. Conclusion Multidrug resistance, biofilm and efflux pump capabilities in Pseudomonas aeruginosa have serious public health implications in the management of infections caused by this organism.

scholarly journals EVALUATION OF EFFLUX PUMP ACTIVITY AND BIOFILM FORMATION IN MULTIDRUG RESISTANT KLEBSIELLA PNEUMONIAE ISOLATES IN TANTA, EGYPT

2021 ◽  
Vol 12 (3) ◽  
pp. 1-5
Author(s):  
Tarek El-Said El-Banna ◽  
Fatma Ibrahim Sonbol ◽  
Heba M El-Dawy ◽  
Lamiaa A Al-Madboly
Keyword(s):  
Drug Resistance ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Treatment Options ◽  
Multidrug Resistant ◽  
High Morbidity ◽  
Biochemical Tests ◽  
Antibiotic Resistance Profile ◽  
Pump Activity

Nosocomial and community acquired infections that caused by multidrug-resistant (MDR) Klebsiella pneumoniae isolates are widespread recently resulting in high morbidity and mortality due to limited number of treatment options with effective antibiotics. The aim of this study is to evaluate the antibiotic resistance profile, biofilm formation and efflux pump activity of MDR K. pneumoniae isolates collected from different hospitals in Tanta, Egypt. A total of 70 K. pneumoniae isolates characterized by standard biochemical tests and confirmed by MALDI-TOF/MS were screened for antibiotic susceptibility, efflux pump activity and biofilm formation. Isolates displayed high resistance to penicillins, cephalosporins, trimethoprim-sulfamethoxazole and the majority of tested fluoro/-quinolones and decreased resistance to imipenem, amikacin, chloramphenicol, tigecycline and colistin. Out of 70 K. pneumoniae isolates, 2 isolates exhibited Pan Drug-Resistance (PDR) profile while 57 (81.4%) and 11 (15.7%) exhibited MDR and Extensively drug-resistance (XDR) profiles, respectively. Sixty-four (91.4%) isolates exhibited efflux pump activity while all tested isolates had the ability to form biofilm with varied degrees as 40 (57.1%), 26 (37.1%), and 4 (5.7%) isolates were strong, moderate and weak biofilm producers, respectively. Also, a strong relation between efflux pump activity and biofilm formation per isolate was detected. In conclusion, Multidrug resistance, biofilm formation and efflux pump capabilities in K. pneumoniae have serious public health implications in the management and control of infections caused by this bacterium. Therefore, a multifaceted approach and precise planning are recommended in controlling these infections

scholarly journals Genome Analysis of Antimicrobial Resistance Genes and Virulence Factors in Multidrug-Resistant Campylobacter fetus Subspecies Isolated from Sheath Wash

2020 ◽  
Vol 10 (4) ◽  
pp. 465-480
Author(s):  
Mpinda Edoaurd Tshipamba ◽  
Ngoma Lubanza ◽  
Mulunda Mwanza
Keyword(s):  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Efflux Pump ◽  
Resistance Mechanism ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Genetic Profile ◽  
Multidrug Efflux Pump ◽  
Campylobacter Fetus ◽  
Antimicrobial Resistance Genes

Campylobacter fetus subspecies are mostly characterized by reproductions problems in cattle and sheep. This study aimed to study the genetic profile and assess the genes mechanism of resistance and their virulence factors using genome sequence analysis. A total of 59 confirmed Campylobacter fetus subspecies based on molecular assays and DNA sequencing were subjected to antimicrobial susceptibility test against 14 antibiotic agents representing the five classes of antibiotics using the disc diffusion method. In addition, sequencing the genome of all strains induced complete resistance against all tested antibiotics. The results of the antimicrobial test indicated that 54.4% had a resistance profile, 26.3% were intermediate, while 19.3% were observed to be susceptible. The Whole Genome Sequencing (WGS) result revealed the presence of different genes, such as Broad-specificity multidrug efflux pump and 16S rRNA (guanine527-N7)-methyltransferase (gidB), efflux pump conferring antibiotic resistance (MacA and MacB), protein-altering cell wall charge conferring antibiotic resistance (PgsA), which have never been reported in Campylobacter fetus subspecies. The WGS also revealed the presence of genes that involved in colonization, adhesion, motility, and invasion, such as type IV secretion system protein (VirD4), S-Layer, cytolethal distending toxin (A, B, and C), Campylobacter invasion antigen (CiaB), and fic domain protein (fic) were among important CDS. The presence of these uncommon genes explains the resistance of Campylobacter fetus subspecies against different tested antibiotics. The results of this study can be used to implement molecular surveillance of Campylobacter fetus subspecies and conduct further studies on the resistance mechanism in these subspecies.

scholarly journals Phenotypic characterization of multidrug-resistant Pseudomonas aeruginosa strains isolated from pediatric patients associated to biofilm formation

2015 ◽  
Vol 172 ◽  
pp. 68-78 ◽  
Author(s):  
Sara A. Ochoa ◽  
Ariadnna Cruz-Córdova ◽  
Gerardo E. Rodea ◽  
Vicenta Cázares-Domínguez ◽  
Gerardo Escalona ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Pediatric Patients ◽  
Multidrug Resistant ◽  
Phenotypic Characterization

scholarly journals Alkylaminoquinolines inhibit the bacterial antibiotic efflux pump in multidrug-resistant clinical isolates

2003 ◽  
Vol 376 (3) ◽  
pp. 801-805 ◽  
Author(s):  
Monique MALLÉA ◽  
Abdallah MAHAMOUD ◽  
Jacqueline CHEVALIER ◽  
Sandrine ALIBERT-FRANCO ◽  
Pierre BROUANT ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Microbial Pathogens ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
High Level ◽  
Antibiotic Efflux

Over the last decade, MDR (multidrug resistance) has increased worldwide in microbial pathogens by efflux mechanisms, leading to treatment failures in human infections. Several Gram-negative bacteria efflux pumps have been described. These proteinaceous channels are capable of expelling structurally different drugs across the envelope and conferring antibiotic resistance in various bacterial pathogens. Combating antibiotic resistance is an urgency and the blocking of efflux pumps is an attractive response to the emergence of MDR phenotypes in infectious bacteria. In the present study, various alkylaminoquinolines were tested as potential inhibitors of drug transporters. We showed that alkylaminoquinolines are capable of restoring susceptibilities to structurally unrelated antibiotics in clinical isolates of MDR Gram-negative bacteria. Antibiotic efflux studies indicated that 7-nitro-8-methyl-4-[2´-(piperidino)ethyl]aminoquinoline acts as an inhibitor of the AcrAB–TolC efflux pump and restores a high level of intracellular drug concentration. Inhibitory activity of this alkylaminoquinoline is observed on clinical isolates showing different resistance phenotypes.

scholarly journals Antistaphylococcal and Antibiotic Resistance Modulatory Activities of Thirteen Cameroonian Edible Plants against Resistant Phenotypes

2018 ◽  
Vol 2018 ◽  
pp. 1-12
Author(s):  
Brice E. N. Wamba ◽  
Armelle T. Mbaveng ◽  
Paul Nayim ◽  
Joachim K. Dzotam ◽  
Ornella J. T. Ngalani ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Antibacterial Activities ◽  
Multidrug Resistant ◽  
Edible Plants ◽  
Bacterial Strains ◽  
Efflux Pump Inhibitor ◽  
Dacryodes Edulis

Background. In this study, 18 methanol extracts from Cameroonian edible plants were tested for their antibacterial activities against 26 strains ofS. aureus; the role of efflux pumps in the resistance of tested bacteria and the antibiotic resistance-modulating activities against selected multidrug-resistant (MDR) phenotypes were also investigated.Methods. Broth microdilution assay was used to evaluate the antibacterial activity, the role of efflux pumps, and the antibiotic resistance-modulating effects of plant extracts.Results. Extracts fromDacryodes edulisseeds (DES) andDacryodes edulisbark (DEB) were active against all 26 tested bacterial strains, within the minimal inhibitory concentration (MIC) range of 256–1024 µg/mL. MIC values varied from 64 to 1024 µg/mL against 96.2% of the 26 tested bacteria forPhaseolus vulgarisleaves (PVL), 92.3% forAzadirachta indicabark (AIB),Dacryodes edulisleaves (DEL), andRicinodendron heudelotiileaves (RHL). The lowest MIC value of 64 µg/mL was obtained with the extract fromCucurbita maximabeans (CMB) against MRSA4 strain and fromUapaca guineensisbark (UGB) against MRSA9 strain. Bacterial efflux pump inhibitor (EPI), carbonyl cyanidem-chlorophenyl hydrazone (CCCP), improved the activity of DES and UGB as well as that of extracts fromHibiscus esculentusleaves (HEL) andUapaca guineensisleaves (UGL) against resistantS. aureusstrains. Antibiotic-modulating effects against more than 70% of theS. aureusstrains tested were obtained when RHL (at MIC/2) was combined with CIP, ERY, and KAN (88.89%), CHL (88.89%), TET (77.78%), and STR (88.89%).Conclusion. The present study demonstrated that the 13 tested plants had antistaphylococcal effects and that DES, HEL, UGL, and UGB could be used in combination with EPI to combat resistance toStaphylococcus aureus. Also, it demonstrated that some studied extracts and mostly RHL could be used as antibiotic resistance modulators to fight against resistant strains ofS. aureus.

scholarly journals Detection of OqxAB and QepA Efflux Pumps and Their Association with Antibiotic Resistance in Klebsiella pneumoniae Isolated From Urinary Tract Infection

2020 ◽  
Vol 7 (4) ◽  
Author(s):  
Manijeh Dehnamaki ◽  
Maryam Ghane ◽  
Laleh Babaeekhou
Keyword(s):  
Urinary Tract Infection ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Tract Infection ◽  
Klebsiella Pneumoniae ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Diffusion Method ◽  
Beta Lactam ◽  
Disk Diffusion Method

Background: The emergence and spread of drug resistance among Klebsiella pneumoniae clinical isolates have limited the treatment options for these bacteria. Efflux pumps are considered as one of the key mechanisms of antibiotic resistance in K. pneumoniae isolates. Objectives: The present study aimed to detect oqxA, oqxB, and qepA efflux genes in K. pneumoniae isolated from urinary tract infection (UTI) and survey their association with antibiotic resistance. Methods: In total, 100 K. pneumoniae isolates were obtained from urine samples, and an antimicrobial susceptibility test was conducted using the disk diffusion method according to the Clinical and Laboratory Standards Institute (CLSI) instructions. Polymerase chain reaction (PCR) was done for the detection of efflux pump genes including, oqxA, oqxB, and qepA, and their association was statistically analyzed with resistance to antibiotics. Results: The highest rate of resistance was obtained against trimethoprim-sulfamethoxazole (72%), amikacin (70%), levofloxacin (68%), gentamicin (56%), ceftazidime (56%), and ceftriaxone (51%), and the lowest resistance was against imipenem (10%). Thirty one percent of isolates were multidrug resistant (MDR). Molecular distribution test showed that 57% and 56% of isolates carried the oqxA and oqxB genes, respectively. Also, the frequency of qepA genes was 21%. The presence of oqxA/oqxB and qepA efflux genes were significantly associated with fluoroquinolone and beta-lactam resistance phenotypes (P < 0.05). Conclusions: The high frequency of efflux genes showed that this resistance mechanism is the main way, along with other resistance mechanisms in K. pneumoniae isolates. It is necessary to adopt appropriate treatment to reduce the incidence of resistance.

scholarly journals Pattern of antibiotics resistance and phenotypic characterization of Multidrug resistant bacteria isolates in four hospitals of Littoral region, Cameroon

2021 ◽  
Vol 11 (1-s) ◽  
pp. 20-30
Author(s):  
Merlin Jonas Wandji Takemegni ◽  
Assob Nguedia Jules clement ◽  
Ateudjieu Jérôme ◽  
Enow Orock George ◽  
Mbopi-Keou Francois-Xavier ◽  
...  
Keyword(s):  
Multidrug Resistant ◽  
Susceptibility Test ◽  
Diffusion Method ◽  
Phenotypic Characterization ◽  
Military Hospital ◽  
Resistant Bacteria ◽  
Antibiotics Resistance ◽  
Multidrug Resistant Bacteria ◽  
Littoral Region

Objective: To describe the pattern of antibiotics resistance and phenotypic characterization of Multidrug resistant bacteria isolates in four hospitals of Littoral region, Cameroon. Methods: We conducted a descriptive hospital based cross-sectional study from December 2018 to May 2019. A simple random sampling was used to swap 10 selected equipment and 10 materials in the mornings after disinfection but before the start of work in seven units. After inoculation in four agar media consecutively (Eosine Metyleine blue, Cled, Manitol salt agar and blood agar ) and incubated in appropriate conditions, the Kirby-Bauer disk-diffusion method was used for antimicrobial susceptibility test. Results: Among 50.4% (119/236) showed positive bacteria growth, a total of 89 (13 species), predominant bacteria and those more likely to cause nosocomial infections were selected and tested each one to 18 antibiotics. There was high level of resistance to Penicillin (amoxicillin (77.5%) and Oxacillin (76.4%)), followed by 3G Cephalosporine (Ceftazidime (74.2%)) and Monobactam (Aztreonam (70.8%)). Although the least level of resistance was observed in Carbapenem (imipenem (5.6%)). The overall prevalence of MDRB was 62.9% (56/89). MRSA were the mostly detected 57.5 % (30/89), followed by ESBL 10.1% (9/89). Military hospital of Douala and Emergency unit was the MDRBs dominantly contaminated area respectively 39.3% (22/56) and 17.9% (10/56). Conclusion: MDRB occurred to be a current public health problem as well as hospital surfaces are worrying reservoir that can be spread to patient, health professionals and visitors. Keywords: Antibiotic resistance, susceptibility test, multidrug-resistant bacteria, Hospital facilities, Units, Littoral Region-Cameroon

scholarly journals Phenotypic Characterization of Extended-Spectrum Beta-Lactamases and Metallo-Beta-Lactamase of Multi Drug Resistant Acinetobacter baumannii Causing Nosocomial Infections in Erbil City

2020 ◽  
Vol 30 (4) ◽  
pp. 51
Author(s):  
Sakar Bakr Smail ◽  
Kamal I. AL-OTRACHI
Keyword(s):  
Acinetobacter Baumannii ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Phenotypic Characterization ◽  
Multi Drug Resistance ◽  
Beta Lactamase ◽  
Beta Lactamases ◽  
Extended Spectrum ◽  
Extended Spectrum Beta Lactamases

Background: Resistance to broad‑spectrum beta‑lactams, mediated by extended‑spectrum beta‑lactamases, and metallo‑beta‑lactamase enzymes, is an increasing problem worldwide. The main aim is to study phenotypic characterization of extended‑spectrum beta‑lactamases and metallo‑beta‑lactamase multidrug resistant Acinetobacter baumannii in Erbil City. Materials and Methods: A total of 112 Acinetobacter baumannii isolations were collected from patients of all age groups from clinical specimens sputum, blood, pus, wound swab, urine and body fluids (Pleural fluid and cerebrospinal fluid) collected from different medical wards and intensive care unit departments of hospitals in Erbil City for a period of one year from march 2018--march 2019. Isolates were tested for the presence of extended‑spectrum beta‑lactamases and metallo‑beta‑lactamase. Detection of extended‑spectrum beta‑lactamases was done by the combined disk diffusion method according to Clinical and Laboratory Standards Institute guidelines, while metallo‑beta‑lactamase was detected by meropenem and imipenem combined with ethylenediaminetetraacetic acid disk method. Results: 25% (28) Acinetobacter baumannii isolates were positive for extended‑spectrum beta‑lactamases, while 100 % (112) were metallo‑beta‑lactamase producers. Conclusion: Acinetobacter baumannii is becoming a global medical challenge due to the emergence of multi-drug resistance. Newer beta lactamase is a matter of concern as they are developing rapidly and lead to treatment failure. Carbapenems are known to be effective therapeutic agents for Acinetobacter baumannii infections and its resistance limits the use to polymyxins and colistin. Several new medicines are still in research and combination of drug therapy is being currently used in the hospitals together with ours to treat multidrug resistant Acinetobacter baumannii infections.

scholarly journals Bifunctional Enzyme SpoT Is Involved in Biofilm Formation ofHelicobacter pyloriwith Multidrug Resistance by Upregulating Efflux Pump Hp1174 (gluP)

2018 ◽  
Vol 62 (11) ◽  
Author(s):  
Xiaoran Ge ◽  
Yuying Cai ◽  
Zhenghong Chen ◽  
Sizhe Gao ◽  
Xiwen Geng ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Multidrug Resistant ◽  
Major Facilitator Superfamily ◽  
Bifunctional Enzyme ◽  
Content Type ◽  
H Pylori ◽  
Major Facilitator

ABSTRACTThe drug resistance ofHelicobacter pyloriis gradually becoming a serious problem. Biofilm formation is an important factor that leads to multidrug resistance (MDR) in bacteria. The ability ofH. pylorito form biofilms on the gastric mucosa is known. However, there are few studies on the regulatory mechanisms ofH. pyloribiofilm formation and multidrug resistance. Guanosine 3′-diphosphate 5′-triphosphate and guanosine 3′,5′-bispyrophosphate [(p)ppGpp] are global regulatory factors and are synthesized inH. pyloriby the bifunctional enzyme SpoT. It has been reported that (p)ppGpp is involved in the biofilm formation and multidrug resistance of various bacteria. In this study, we found that SpoT also plays an important role inH. pyloribiofilm formation and multidrug resistance. Therefore, it was necessary to carry out some further studies regarding its regulatory mechanism. Considering that efflux pumps are of great importance in the biofilm formation and multidrug resistance of bacteria, we tried to determine whether efflux pumps controlled by SpoT participate in these activities. We found that Hp1174 (glucose/galactose transporter [gluP]), an efflux pump of the major facilitator superfamily (MFS), is highly expressed in biofilm-forming and multidrug-resistant (MDR)H. pyloristrains and is upregulated by SpoT. Through further research, we determined thatgluPis involved inH. pyloribiofilm formation and multidrug resistance. Furthermore, the average expression level ofgluPin the clinical MDR strains (C-MDR) was considerably higher than that in the clinical drug-sensitive strains (C-DSS). Taken together, our results revealed a novel molecular mechanism ofH. pyloriresistance to multidrug exposure.

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