Biofilm Formation and Antimicrobial Resistance Pattern of Uropathogenic E. Coli : Comparison of Phenotypic and Molecular Methods

2017 ◽  
Vol 26 (2) ◽  
pp. 37-45 ◽  
Author(s):  
Amal F. Makled ◽  
Eman H. Salem ◽  
Asmaa M. Elbrolosy
2020 ◽  
Vol 13 (2) ◽  
pp. 360-363
Author(s):  
Shikha Tamta ◽  
Obli Rajendran Vinodh Kumar ◽  
Shiv Varan Singh ◽  
Bommenahalli Siddaramiah Pruthvishree ◽  
Ravichandran Karthikeyan ◽  
...  

Background and Aim: Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli are gradually increasing worldwide and carry a serious public threat. This study aimed to determine the antimicrobial resistance pattern of ESBL-producing E. coli isolated from fecal samples of piglets and pig farm workers. Materials and Methods: Fecal samples from <3-month-old piglets (n=156) and farm workers (n=21) were processed for the isolation of ESBL-producing E. coli in MacConkey agar added with 1 μg/mL of cefotaxime. E. coli (piglets=124; farm workers=21) were tested for ESBL production by combined disk method and ESBL E-strip test. Each of the ESBL-positive isolate was subjected to antibiotic susceptibility testing. The ESBL-producing E. coli were further processed for genotypic confirmation to CTX-M gene. Results: A total of 55 (44.4%, 55/124) and nine (42.9%, 9/21) ESBL-producing E. coli were isolated from piglets and farm workers, respectively. Antibiotic susceptibility testing of the ESBL-positive E. coli isolates from piglets and farm workers showed 100% resistance to ceftazidime, cefotaxime, cefotaxime/clavulanic acid, ceftazidime/clavulanic acid, and cefpodoxime. A proportion of 100% (55/55) and 88.9% (8/9) ESBL-positive E. coli were multidrug resistance (MDR) in piglets and farm workers, respectively. On genotypic screening of the ESBL E. coli isolated from piglets (n=55), 15 were positive for the blaCTX-M gene and of the nine ESBL E. coli from farm workers, none were positive for the blaCTX-M gene. Conclusion: Although there was no significant difference in isolation of ESBL-producing E. coli between piglets and farm workers, the ESBL-positive E. coli from piglets showed relatively higher MDR than farm workers.


Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 420 ◽  
Author(s):  
Mst. Sonia Parvin ◽  
Sudipta Talukder ◽  
Md. Yamin Ali ◽  
Emdadul Haque Chowdhury ◽  
Md. Tanvir Rahman ◽  
...  

Escherichia coli is known as one of the most important foodborne pathogens in humans, and contaminated chicken meat is an important source of foodborne infection with this bacterium. The occurrence of extended-spectrum β-lactamase (ESBL)-producing E. coli (ESBL-Ec), in particular, in chicken meat is considered a global health problem. This study aimed to determine the magnitude of E. coli, with special emphasis on ESBL-Ec, along with their phenotypic antimicrobial resistance pattern in frozen chicken meat. The study also focused on the determination of ESBL-encoding genes in E. coli. A total of 113 frozen chicken meat samples were purchased from 40 outlets of nine branded supershops in five megacities in Bangladesh. Isolation and identification of E. coli were done based on cultural and biochemical properties, as well as PCR assay. The resistance pattern was determined by the disc diffusion method. ESBL-encoding genes were determined by multiplex PCR. The results showed that 76.1% of samples were positive for E. coli, of which 86% were ESBL producers. All the isolates were multidrug-resistant (MDR). Resistance to 9–11 and 12–13 antimicrobial classes was observed in 38.4% and 17.4% isolates, respectively, while only 11.6% were resistant to 3–5 classes. Possible extensive drug resistance (pXDR) was found in 2.3% of isolates. High single resistance was observed for oxytetracycline (93%) and amoxicillin (91.9%), followed by ampicillin (89.5%), trimethoprim–sulfamethoxazole, and pefloxacin (88.4%), and tetracycline (84.9%). Most importantly, 89.6% of isolates were resistant to carbapenems. All the isolates were positive for the blaTEM gene. However, the blaSHV and blaCTX-M-2 genes were identified in two ESBL-non producer isolates. None of the isolates carried the blaCTX-M-1 gene. This study provided evidence of the existence of MDR and pXDR ESBL-Ec in frozen chicken meat in Bangladesh, which may pose a risk to human health if the meat is not properly cooked or pickled raw only. This emphasizes the importance of the implementation of good slaughtering and processing practices by the processors.


2019 ◽  
Vol 11 (01) ◽  
pp. 017-022 ◽  
Author(s):  
Rashmi M. Karigoudar ◽  
Mahesh H. Karigoudar ◽  
Sanjay M. Wavare ◽  
Smita S. Mangalgi

Abstract BACKGROUND: Escherichia coli accounts for 70%–95% of urinary tract infections (UTIs). UTI is a serious health problem with respect to antibiotic resistance and biofilms formation being the prime cause for the antibiotic resistance. Biofilm can restrict the diffusion of substances and binding of antimicrobials. In this context, the present study is aimed to perform in vitro detection of biofilm formation among E. coli strains isolated from urine and to correlate their susceptibility pattern with biofilm formation. MATERIALS AND METHODS: A total of 100 E. coli strains isolated from patients suffering from UTI were included in the study. The identification of E. coli was performed by colony morphology, Gram staining, and standard biochemical tests. The detection of biofilm was carried out by Congo Red Agar (CRA) method, tube method (TM), and tissue culture plate (TCP) method. Antimicrobial sensitivity testing was performed by Kirby–Bauer disc diffusion method on Muller–Hinton agar plate. RESULTS: Of the 100 E. coli strains, 49 (49%) and 51 (51%) were from catheterized and noncatheterized patients, respectively. Biofilm production was positive by CRA, TM, and TCP method were 49 (49%), 55 (55%), and 69 (69%), respectively. Biofilm producers showed maximum resistance to co-trimoxazole (73.9%), gentamicin (94.2%), and imipenem (11.6%) when compared to nonbiofilm producers. Significant association was seen between resistance to antibiotic and biofilm formation with a P = 0.01 (<0.05). CONCLUSION: A greater understanding of biofilm detection in E. coli will help in the development of newer and more effective treatment. The detection of biofilm formation and antibiotic susceptibility pattern helps in choosing the correct antibiotic therapy.


2007 ◽  
Vol 74 (2) ◽  
pp. 391-395 ◽  
Author(s):  
Artashes R. Khachatryan ◽  
Thomas E. Besser ◽  
Douglas R. Call

ABSTRACT Association of specific antimicrobial resistance patterns with unrelated selective traits has long been implicated in the maintenance of antimicrobial resistance in a population. Previously we demonstrated that Escherichia coli strains with a specific resistance pattern (resistant to streptomycin, sulfadiazine, and tetracycline [SSuT]) have a selective advantage in dairy calf intestinal environments and in the presence of a milk supplement commonly fed to the calves. In the present study we identified the sequence of the genetic element that confers the SSuT phenotype and show that this element is present in a genetically diverse group of E. coli isolates, as assessed by macrorestriction digestion and pulsed-field gel electrophoresis. This element was also found in E. coli isolates from 18 different cattle farms in Washington State. Using in vitro competition experiments we further demonstrated that SSuT strains from 17 of 18 farms were able to outcompete pansusceptible strains. In a separate set of experiments, we were able to transfer the antimicrobial resistance phenotype by electroporation to a laboratory strain of E. coli (DH10B), making that new strain more competitive during in vitro competition with the parental DH10B strain. These data indicate that a relatively large genetic element conferring the SSuT phenotype is widely distributed in E. coli from cattle in Washington State. Furthermore, our results indicate that this element is responsible for maintenance of these traits owing to linkage to genetic traits that confer a selective advantage in the intestinal lumens of dairy calves.


2015 ◽  
Vol 13 (4) ◽  
pp. 1020-1028 ◽  
Author(s):  
Stefanie Heß ◽  
Claudia Gallert

Tolerance of antibiotic susceptible and antibiotic resistant Escherichia coli, Enterococcus and Staphylococcus strains from clinical and wastewater samples against ozone was tested to investigate if ozone, a strong oxidant applied for advanced wastewater treatment, will affect the release of antibiotic resistant bacteria into the aquatic environment. For this purpose, the resistance pattern against antibiotics of the mentioned isolates and their survival after exposure to 4 mg/L ozone was determined. Antibiotic resistance (AR) of the isolates was not correlating with higher tolerance against ozone. Except for ampicillin resistant E. coli strains, which showed a trend towards increased resistance, E. coli strains that were also resistant against cotrimoxazol, ciprofloxacin or a combination of the three antibiotics were similarly or less resistant against ozone than antibiotic sensitive strains. Pigment-producing Enterococcus casseliflavus and Staphylococcus aureus seemed to be more resistant against ozone than non-pigmented species of these genera. Furthermore, aggregation or biofilm formation apparently protected bacteria in subsurface layers from inactivation by ozone. The relatively large variance of tolerance against ozone may indicate that resistance to ozone inactivation most probably depends on several factors, where AR, if at all, does not play a major role.


2018 ◽  
Vol 16 (2) ◽  
pp. 178-183
Author(s):  
Dhiraj Shrestha ◽  
Pratigya Thapa ◽  
Dinesh Bhandari ◽  
Hiramani Parajuli ◽  
Prakash Chaudhary ◽  
...  

Background: The study was designed to provide account of etiological agents of urinary tract infection in pediatric patients and the antimicrobial resistance pattern plus biofilm producing profile of the isolates.Methods: The prospective study was conducted in Alka Hospital, Nepal with 353 clean catch urine samples from children. It was obtained during July 2014 to January 2015 which were first cultured by semi-quantitative method, followed by antimicrobial susceptibility testing and biofilm production assay on Congo red agar. Multidrug- resistance, extensively drug- resistance and pandrug- resistance among isolates were considered as per international consensus.Results: Out of 353 samples, 64 (18.13%) showed positive growth in culture, confirming urinary tract infection. E. coli, 44 (68.8%) was the predominant organism followed by Klebsiella spp. 6 (14.1%). Most E. coli were sensitive to amikacin (93.2%) followed by nitrofurantoin (86.4%), and highly resistant to ampicillin (95.5%). Of 64 isolates, 23 (35.93%) were found to be multidrug- resistant strains. Biofilm was produced by 36 (56.25%) isolates.Conclusions: This study showed higher biofilm production and resistance to in-use antibiotics rendering ineffective for empirical use. Regular surveillance of resistance patterns should be done to regulate multidrug- resistant bugs and to ensure effective management of urinary tract infection in children in a tertiary care setups.Keywords: AMR; antimicrobial resistance; biofilm; urinary tract infection; UTI.


Author(s):  
Gaurab Risal ◽  
Aayush Shrestha ◽  
Saroj Kunwar ◽  
Gajal Paudel ◽  
Rameshwor Dhital ◽  
...  

Background: In urinary tract infections, an important role is played by bacterial biofilms which are responsible for persistence infections together with the antimicrobial resistance. Higher resistance can be seen in biofilm forming uropathogens in comparison with free-floating bacteria. So, the present study was performed with a goal to find the prevalence of biofilm formation and also the antimicrobial resistant pattern of uropathogens.Methods: A descriptive method was conducted at Modern Technical College, Sanepa, Lalitpur in samples isolated from UTI suspected patients. The overall duration of this study was approximately 3 months. Total of 50 isolated E. coli was tested for biofilm formation and antimicrobial susceptibility testing was done by Kirby-Bauer disc diffusion method on Mueller Hinton agar as per CLSI guidelines.Results: From the 50 isolates of E. coli, 32 were biofilm producers (3 strong and 29 moderate) and 18 were weak/non-biofilm producers. Among the biofilm producers, cefotaxime was more resistant in 20 of the isolates followed by ceftriaxone in 16 and amoxyclav in 13, whereas amikacin was least resistant in 2 of the isolates.Conclusions: Among the isolated E. coli, biofilm-forming isolates showed higher antimicrobial resistance as compared to the non-biofilm producer. Thus, uropathogen should be routinely screened for biofilm formation. 


Author(s):  
Mallikarjun Gundappa ◽  
C. Prabhurajeshwar ◽  
Sarfaraz Ahmed ◽  
H.M. Navya ◽  
M. Vijayasarathy ◽  
...  

Background: The present study was undertaken to study the percentage of pathogenic bacteria present in different fruit and vegetable samples available in the market in and around Kalaburagi, Karnataka, South India. Methods: A total of 940 different samples were collected from Kalaburagi, out of which 390 (23.84%) were fruits and 550 (35.27%) were vegetables. The pathogenic bacteria were isolated, by enrichment culture method using peptone water. The bacterial isolates were identified by convention microbial identification procedures. Result: Antibiotic resistant testing by disc diffusion method performed for E.coli, Salmonella and Shigella. Among the pathogens, E. coli (86.50%) of the isolates were resistant to Nalidixic acid while Imipenem and Trimethoprim-Sulfamethoxazole has the lowest resistance (19.84%), Salmonella (86.66%) isolates were resistant to Ciprofloxacin while Norfloxacin has the lowest resistance (4.76%) and Shigella (80.35%) isolates were resistant to Vancomycin while Amoxicillin has the lowest resistant (3.57%). Multiple drug resistance (MDR) was seen in E. coli at (38.88%), Salmonella at (26.66%) and Shigella at (10.71%) accordingly. The study therefore shown that E.coli, Salmonella and Shigella occur in Fruits and vegetables which collected from market place in Kalaburagi, Karnataka India, As per the results, adequate precaution should be taken while handles these fruits and vegetables. The antimicrobial resistance pattern shown by the isolates is an indication that adequate measurement needs to be taken to regulate the drug use in both humans and animals in order to minimize the risk of increasing antimicrobial resistance.


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