Effects of Lactobacillus acidophilus and Lactobacillus helveticus on biofilm formation by ESBL-producing uropathogenic Escherichia coli

2020 ◽  
Vol 29 (3) ◽  
pp. 163-170
Author(s):  
Alaa M. Ibrahim ◽  
Magda I. Ayoub ◽  
Alaa R. Awad

Background: Uropathogenic E. coli (UPEC) are important clinical pathogens and are considered as one of the major causes of urinary tract infection. Unfortunately, the treatment of UPEC infections is becoming more difficult because isolates are increasingly resistant to commonly used antimicrobial agents. The potential ability of UPEC to form biofilm might explain its outstanding antibiotic resistance and survival properties. Objectives: We aimed to assess the effect of cell free supernatants (CFS) of Lactobacillus acidophilus La-5 and Lactobacillus helveticus B-734 on the ability of ESBL-producing UPEC isolates to form biofilm in vitro and to eradicate the already formed biofilm of UPEC isolates. Methodology: A total of 50 isolates of ESBLproducing UPEC, were recovered from different private laboratories, then subcultured on MacConkey’s medium and identified by conventional microbiological methods. Antibiotic susceptibility testing was performed by Kirby-Bauer disc diffusion method and confirmed for ESBL production. The ability of UPEC to produce biofilm was determined using Congo red agar (CRA) method and tissue culture plate (TCP) method. CFS of L.acidophilus La-5 and L. helveticus B-734 were prepared and used to assess its effect on the ability of ESBL-producing UPEC isolates to form biofilm in vitro and the eradicationof the already formed biofilm. Results: Biofilm formation was found in 28% of theisolates. Furthermore, the strong biofilm-forming category was detected in 10% of the isolates, whereas the moderate biofilm-forming category was detected in 18%. CFS of L. acidophilus La-5 was more effective (91.12%) in inhibiting the formation of E. coli biofilm than CFS of L. helveticus B-734 (76.44%), while it was almost the same efficacy (83%) regarding eradication of E. coli biofilm. All ESBL- producing UPEC isolates were sensitive to cefoxitin (100%) followed by nitrofurantoin (98%) and imipenem (94%). Conclusion: L. acidophilus La-5 and L. helveticus B-734 may be a new therapeutic options for UTI caused by UPEC.

2021 ◽  
Vol 2 (2) ◽  
pp. 7-13
Author(s):  
PS Utulo ◽  
EU Umeh ◽  
GM Gberikon ◽  
PO Abba

Beta-lactamase production by Escherichia coli and other bacteria is one of the most important mechanism of resistance to beta-lactam antibiotics such as Penicillins and Cephalosporins which are the most commonly used antibiotics in the treatment of bacterial infections in Hospitals. Resistance to antibiotics is emerging worldwide as a threat to human health resulting in limitations of therapeutic options. Susceptibility tests of microorganisms to antimicrobial agents do not reveal production of Extended-Spectrum ß-lactamase, hence the need to detect their presence by phenotypic or molecular techniques. This study is aimed at determining the prevalence of ESBL-production by Escherichia coli isolates from clinical samples collected from some selected health facilities in Otukpo, using phenotypic detection. Four hundred (400) specimens were collected from four different health facilities in Otukpo. Specimens collected included urine, blood, stool and wound swabs from 222 females and 178 males. Standard microbiological methods were employed for isolation, identification and characterization of Escherichia coli isolates from these specimens. Antibiotic susceptibility of the E. coli isolates was determined using Kirby-Bauer disc diffusion method while double disc synergy test (DDST) was used for ESBL production. Sixty-eight, (17%) of the 400 specimens tested for ESBL production were positive. Isolates from stool 22(22.7%) had the highest prevalence of ESBL-producing E. coli, followed by isolates from blood 4(21.4%). Urine isolates had the least prevalence 31(13.8%). Isolates from female subjects, 38(17.1%) had higher positivity rate than their male counterpart 30(16.9%). Otukpo General Hospital had the highest prevalence 20(20.0%) of ESBL followed by Otia Hospital 18(18.0%). Comprehensive health center, Otukpo had the least prevalence 14(14.0%). There is a high prevalence of ESBL production by Escherichia coli isolated from health facilities in Otukpo. Routine drug resistance surveillance therefore has become necessary to guide the appropriate and judicious antibiotic use.


2021 ◽  
Vol 74 (9) ◽  
pp. 2109-2111
Author(s):  
Evheniia A. Shtaniuk ◽  
Oleksandra O. Vovk ◽  
Larisa V. Krasnikova ◽  
Yuliia I. Polyvianna ◽  
Tetiana I. Kovalenko

The aim: Study of antibacterial activity of the preparations, containing antiseptic dioxidine and antibiotic levofloxacin in vitro on standard strains of main optional-anaerobic pathogens of purulent-inflammatory processes of surgical wounds S. aureus, E. coli, P. aeruginosa and definition of more effective ones on them. Materials and methods: Solutions of dioxidine 1.2 %, dioxidine 1.2% with decamethaxin, Dioxisole, water soluble ointment with dioxidine 1.2% and levofloxacin 0.1% with decamethaxin were used in experiment. Antibacterial activity was studied on standard strains of S. aureus АТСС 25923, E. coli АТСС 25922, P. aeruginosa АТСС 27853. Distinguishing and identification of pure cultures of bacteria was done according to generally accepted microbiological methods. Determination of purulent-inflammatory processes pathogens sensitivity was done by disco-diffuse method on Mueller-Hinton medium. Antibacterial activity of solutions and ointments was studied with the help of agar diffusion method (“well” method) according to methodic recommendations. Each investigation was repeated 6 times. Method of variation statistics was used for the research results analysis. Results: All antibacterial preparations under study are effective and highly effective on S. aureus АТСС 25923, E. coli АТСС 25922, P. aeruginosa АТСС 27853. Solution with 1.2 % dioxidine with decamethaxin and ointment with 0.1 % levofloxacin and decamethaxin have larger growth retardation zones towards S. aureus and P. aeruginosa. E. coli strains are more sensitive to the solution of Dioxisole and ointment with 1.2 % dioxidine. Conclusions: All strains are sensitive, most of them are highly sensitive, up to 5 antibacterial preparations under study in vitro.


2020 ◽  
Vol 20 (1) ◽  
pp. 238-247
Author(s):  
Ashraf A Kadry ◽  
Nour M Al-Kashef ◽  
Amira M El-Ganiny

Background: Escherichia coli is the most predominant pathogen involved in UTIs. Mainly, fimbrial surface appendages are impli- cated in adherence to urothelium besides non-fimbrial proteins. Objectives: To determine prevalence of genes encoding fimbrial and non-fimbrial proteins among Uropathogenic Escherichia coli (UPEC). Furthermore, distribution of these genes and biofilm formation capacity were investigated in relation to antimicrobial resistance. Methods: Antimicrobial susceptibility of 112 UPEC isolates was performed using disc diffusion method. ESBL production was confirmed by double disc synergy test. Genes encoding fimbrial and non-fimbrial proteins were detected using PCR and biofilm formation was investigated using microtitre plate assay. Results: UPEC isolates exhibited high resistance against doxycyclines (88.39 %), β-lactams (7.14-86.6%), sulphamethoxaz- ole–trimethoprim (53.75%) and fluoro-quinolones (50%). Fifty percent of tested isolates were ESBL producers. PapGII gene was statistically more prevalent among pyelonephritis isolates. SfaS, focG and picU genes were statistically associated with flu- oro-quinolone (FQs) sensitive isolates and Dr/afaBC gene was statistically associated with ESBL production. Moreover, non- MDR isolates produced sturdier biofilm. Conclusion: PapGII adhesin variant seems to have a critical role in colonization of upper urinary tract. There is a possible link between antimicrobial resistance and virulence being capable of affecting the distribution of some genes besides its negative impact on biofilm formation. Keywords: Urinary tract infection; Escherichia coli; UPEC; adhesin genes; ESBL; biofilm.  


2020 ◽  
Vol 14 (4) ◽  
pp. 2577-2584
Author(s):  
Tariq Ahmad Shah ◽  
P. Preethishree ◽  
Ashwini ◽  
Vidya Pai

Urinary tract infection (UTI) is one of the most common complaints in the outpatient clinic and a major health problem owing to the emergence of antibiotic resistance and biofilm formation. The objective of this study was to isolate and identify the causative bacterial agent of UTI and detect in vitro biofilm formation by Escherichia coli and investigate its correlation with antibiotic resistance. Urine samples from 519 patients with suspected UTIs were collected and processed by conventional microbiological procedures. Antimicrobial susceptibility testing for E. coli isolates was performed on Mueller Hinton agar (MHA) plates using the Kirby-Bauer disk diffusion method. Biofilm production was evaluated using the tissue culture plate method. Of 519 urine samples, 115 (22.1%) showed significant bacteriuria. The most common isolate was E. coli (n=57, 49.6%), followed by Klebsiella spp. (n=23, 20%). All E. coli isolates were evaluated for their ability to form biofilms in vitro. Of 57 isolates, 50 (87.7%) were biofilm producers and 7 (12.3%) were non-biofilm producers. Antibiogram of E. coli isolates revealed the highest resistance to ampicillin (96.5%) and nitrofurantoin (91.2%), followed by amoxyclav (82.5%), ceftazidime (73.7%), cefepime (71.9%), and tetracycline (71.9%). A significant association (p<0.05) was observed between biofilm formation and resistance to amoxyclav, ceftazidime, cefepime, imipenem, and nitrofurantoin. A significant correlation was noted between biofilm production and antibiotic resistance. Hence, screening of all isolates of uropathogenic E. coli for biofilm production and studying their antibiogram would allow appropriate choice of antibiotic therapy.


Author(s):  
V. V. Pantyo ◽  
M. M. Fizer ◽  
O. I. Fizer ◽  
G. M. Koval ◽  
E.M. Danko

Annotation. The development and rapid pace of the spread of resistance to antimicrobial agents predetermines the search for new methods of counteracting pathogenic and conditionally pathogenic microorganisms. In this context, studies of the antimicrobial activity of newly synthesized chemicals, which in the future can be considered as candidates for antiseptic and disinfectants, are relevant. The aim of the work was to determine the antimicrobial activity of new ionic associates based on the surface-active cetylpyridinium cation with respect to certain opportunistic microorganisms. The antimicrobial activity of four ionic associates based on the cetylpyridinium cation with respect to clinical isolates of E. coli, P. vulgaris, K. pneumonia, P. aeruginosa, S. aureus, as well as the collection test strains of S. aureus ATCC 25923, E. coli ATCC 29522 and P. aeruginosa ATCC 27853 was studied. Screening studies were performed by the disk diffusion method. With substances that showed an antimicrobial effect, quantitative studies were carried out by the method of serial macro-dilutions in a liquid nutrient media. Screening studies revealed the antibacterial activity of the substances against E. coli ATCC 25923, E. coli (clinical isolate), P. vulgaris and K. pneumonia. With these microorganisms quantitative studies were carried out with the determination of the minimum inhibitory and minimum bactericidal concentrations. The most pronounced antimicrobial activity for the investigated microflora was shown by tetraphenylborate and cetylpyridinium perchlorate. The MIC and MBC values of these substances ranged between 1.625–3.125 mmol / L and 3.125–12.5 mmol / L, respectively. The studied associates showed high antimicrobial activity against representatives of the Enterobacteriaceae family in in vitro studies. Promising is the further study of the effect of the counter-anion associates of cationic surfactants on the biofilm formation of conditionally pathogenic microorganisms.


2021 ◽  
Vol 10 (7) ◽  
pp. 414-418
Author(s):  
Greeshma Hareendranath

BACKGROUND Escherichia coli is one of the most important causes of urinary tract infections (UTIs). Increased antibiotic resistance may limit the therapeutic options for the treatment of these infections. Fosfomycin trometamol is a phosphonic acid derivative, which acts primarily by interfering with bacterial peptidoglycan synthesis with broad spectrum of activity against agents causing urinary tract infection with good antibiofilm activity and limited reports of resistance and hence is increasingly called upon for the treatment of multi drug resistant (MDR) organisms causing UTI. There are limited studies from India regarding the efficacy of this drug; so, the study was conducted to determine the in vitro efficacy of fosfomycin against uropathogenic MDR E. coli. METHODS This was a prospective study done in the Department of Microbiology, Government T.D. Medical College, Alappuzha, over a period of 1 year from April 2018 to March 2019. A total of 150 MDR urine samples were processed by routine microbiological methods and after identification of E. coli urinary isolates, antibiotic susceptibility testing was performed and results were interpreted following the Clinical and Laboratory Standards Institute guidelines (CLSI). Fosfomycin sensitivity was tested by the Kirby-Bauer disc diffusion method. RESULTS Among the 150 MDR urinary E. coli isolates, 148 (98 %) were sensitive to fosfomycin in our study. The susceptibility rate of fosfomycin was clearly higher than other commonly used drugs for UTI. All extended-spectrum beta-lactamases (ESBL) producing E. coli were sensitive to this drug. The susceptibility for nitrofurantoin was fair, whereas for ampicillin, norfloxacin, cefotaxime and trimethoprim / sulphamethoxazole was found poor. Relatively better rates of resistance were observed for parenteral antibiotics. CONCLUSIONS With an enormous increase in the bacterial pathogens resistant to first-line antibiotics, there has been a revival in the use of fosfomycin. The convenience of a single dose regimen, a good activity proven invitro, and minimal propensity for development of resistance pathogens makes fosfomycin an attractive regimen for the treatment of uncomplicated community and hospital acquired UTIs. In this regard, with the existing limited options for treating MDR organisms, fosfomycin finds its utility acting as an effective and promising option in the treatment of UTIs due to MDR pathogens in the future.


Author(s):  
Lubna Farooq ◽  
Shaikh Nadeem Ahmed ◽  
Muhammad Aitmaud Uddolah Khan ◽  
Akhtar Ali ◽  
Sehrish Mehmood ◽  
...  

Background: Urinary tract infections are found to be commonest bacterial infections across the globe. Various studies have demonstrated high prevalence rate of UTIs in Pakistan. Multiple broad spectrum antibiotics are being used for the treatment of UTI but the resistance by the pathogen against these drugs is increasing worldwide. As the resistance in the organisms is increasing day by day, and it is now hall mark and matter of concern for clinicians to treat uropathogenic E. coli, so there is a pertinent need to explore new sensitive antibiotics or alternative options to manage the disease. Aims: To determine the pathogen burden and susceptibility pattern of ceftolozane/tazobactam against MDR E. coli isolates from clinical specimens of urinary tract infections in Karachi. Study Design:  It was an in-vitro clinical study. Study Settings: The study was conducted in department of Pharmacology, Baqai Medical University and isolates were collected from Microbiology laboratory of Karachi. Methodology: On the basis of identification methods, one hundred and fifty (150) strains of E. coli were isolated from 650 specimen of urine. Clinical isolates were identified by standard and specific microbiological methods. The antibiotic susceptibility pattern was determined by Kirby Bauer Disc diffusion method. Samples were processed as per procedures defined by Clinical and Laboratory Standards Institute (CLSI) guidelines 2018. Results: Out of 150 isolates of E. coli, 95 (63.3%) were MDR E. coli. majority of the cases were obtained from age group 61-80 year (32.6%). Highest sensitivity was seen by ceftolozane/tazobactam (96%) followed by ceftriaxone (88%). Least sensitivity was observed with Imipenem (13.70%). However increased trend of resistance was seen among all empirical used drugs.


Author(s):  
Touhida Ishma ◽  
H. M. Sayeed Uddin ◽  
Anik Paul ◽  
Farahnaaz Feroz ◽  
Mrityunjoy Acharjee

<p class="abstract"><strong>Background:</strong> Practice of hand washing is very important to eliminate the microbial contamination especially during the work in laboratories, hospital and even at home before taking food. Proper use of hand sanitizer can significantly reduce the rate of hospital acquired infection also.</p><p class="abstract"><strong>Methods:</strong> The current investigation was designated to identify different bacterial species from the upper skin of hands of the laboratory managements through conventional culture methods and the efficacy of the samples (Dettol, Purell and Savlon) against the isolated bacteria through agar well diffusion method and minimum inhibitory concentration (MIC)  </p><p class="abstract"><strong>Results:</strong> Different concentrations such as 25%, 50% and 100% of each of antimicrobial agents showed their antibacterial activity against <em>Staphylococcus </em>spp., <em>Klebsiella </em>spp., <em>E. coli</em> and <em>Pseudomonas</em> spp. those were isolated from the hand. 25% of Dettol exhibited 20 mm zone diameter against <em>Klebsiella</em>spp. Whereas 25% of Purell and Savlon unveiled 15 mm and 22 mm zone of inhibition against <em>E. coli</em> and <em>Pseudomonas</em> spp. respectively. In case of 100% sample of Dettol, Purell and Savlon, the highest zone diameter was observed as 55 mm, 50 mm and 45 mm against <em>Klebsiella </em>spp.,<em> Staphylococcus </em>spp., and <em>E. coli</em> consecutively. The MIC of Dettol sample was 8 µl against <em>Staphylococcus </em>spp., and <em>Pseudomonas</em> spp. In case of Purell, the MIC was 128 µl against <em>Staphylococcus</em> spp., <em>Klebsiella </em>spp. and <em>Pseudomonas </em>spp. were inhibited at 4 µl samples.</p><p class="abstract"><strong>Conclusions:</strong> The in-vitro antibacterial activity of the hand sanitizers was so satisfactory against the isolated bacteria. This finding would be very helpful for the laboratory management in order to minimize the rate of contamination during the research and supervision of the different experiment.</p>


2020 ◽  
Vol 11 ◽  
pp. 37-43
Author(s):  
Prof. Teodora P. Popova ◽  
Toshka Petrova ◽  
Ignat Ignatov ◽  
Stoil Karadzhov

The antimicrobial action of the dietary supplement Oxidal® was tested using the classic Bauer and Kirby agar-gel diffusion method. Clinical and reference strains of Staphylococcus aureus and Escherichia coli were used in the studies. The tested dietary supplement showed a well-pronounced inhibitory effect against the microbial strains commensurable with that of the broad-spectrum chemotherapeutic agent Enrofloxacin and showed even higher activity than the broad spectrum antibiotic Thiamphenicol. The proven inhibitory effect of the tested dietary supplement against the examined pathogenic bacteria is in accordance with the established clinical effectiveness standards for antimicrobial agents.


2021 ◽  
pp. 088391152110142
Author(s):  
Velu Gomathy ◽  
Venkatesan Manigandan ◽  
Narasimman Vignesh ◽  
Aavula Thabitha ◽  
Ramachandran Saravanan

Biofilms play a key role in infectious diseases, as they may form on the surface and persist after treatment with various antimicrobial agents. The Staphylococcus aureus, Klebsiella pneumoniae, S. typhimurium, P. aeruginosa, and Escherichia coli most frequently associated with medical devices. Chitosan sulphate from marine litter (SCH-MW) was extracted and the mineral components were determined using atomic absorption spectroscopy (AAS). The degree of deacetylation (DA) of SCH was predicted 50% and 33.3% in crab and shrimp waste respectively. The elucidation of the structure of the SCH-MW was portrayed using FT-IR and 1H-NMR spectroscopy. The molecular mass of SCH-MW was determined with Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF). The teratogenicity of SCH-MW was characterized by the zebrafish embryo (ZFE) model. Antimicrobial activity of SCH-MW was tested with the agar well diffusion method; the inhibitory effect of SCH-MW on biofilm formation was assessed in 96 flat well polystyrene plates. The result revealed that a low concentration of crab-sulfated chitosan inhibited bacterial growth and significantly reduced the anti-biofilm activity of gram-negative and gram-positive bacteria relatively to shrimp. It is potentially against the biofilm formation of pathogenic bacteria.


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