Detection of β-lactamase genes (blaTEM, bla CTX-M and bla SHV) in uropathogens isolated from patients with UTI

2021 ◽  
Vol 30 (3) ◽  
pp. 97-103
Author(s):  
Hazem Hamed ◽  
Asaad Gaber ◽  
Eman M. EL Sehsah

Background: The problem of antibiotic resistance has become worldwide health issue among hospitalized patients in developing countries, where the inappropriate usage of antibiotics is common. The widespread occurrence of multidrug resistant strains (MDRS) of Escherichia coli (E. coli) and klebsiella pneumoniae (K. pneumoniae) is currently considered avital medical challenge. The identification of ESβL genes and their MDRS in widely isolated uropathogens may provide a helpful tool for choosing effective antibiotic therapy and clinical improvement. Objective: The aim of this study is to determine the prevalence of (bla.TEM, bla.CTX-M, and bla.SHV) β-lactamase DNAs in E. coli and K. pneumoniae, and MDRS of uropathogens E. coli and k. pneumoniae isolated from patients urine UTI diagnosed who were admitted to the Urology & Nephrology Center. Methodology: Clinical E. coli and K. pneumoniae isolates were isolated from urine specimens of UTI diagnosed patients. Microbiological techniques were utilised to identify the isolates. Antibiotics susceptibility analysis was done utilising the Vitek2 system (bioMérieux, Marcy l’Etoile, France). The double disc diffusion technique (DDDT) was used for detecting phenotypic ESBL in isolates. Using the traditional PCR technique, the ESBL genes were identified. Results: Antibiotic resistance of E. coli’s isolates to commonly used antibiotics was high (88.2% resistant to Amoxicillin/clavulanate, 83.5 % were resistant to Trimethoprim-sulfamethoxazole) while minimum resistance appeared against Amikacin (AK) 10.5% and Imipenem (IP) 14%. For isolates of K. pneumoniae, the resistance was higher for nitrofurantoin, ceftazidime, cefotaxime, ciprofloxacin 96.6%, 83.3 %,83.3%, 80% respectively .In our study, out of 115 isolates, E. coli and K. pneumoniae isolates, 81 (70.43%) were phenotypically positive for ESβL by Combination Disc Test (CDT). The most commonly molecularly detected ESBL genes were TEM 80% of K. pneumoniae isolates and 77.6% of E. coli isolates .followed by SHV 70% of K. pneumoniae isolates and 34.1% of E. coli isolates and finally, CTXM 67% of isolates of E. coli and 63.3% of isolates of K. pneumoniae. Conclusions: The current study concluded that a high rate of resistance has been developed in uropathogens with empiric antibiotic treatment. It emphasized the alarming role of β-lactamases, especially ESßLs in antibiotic

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Benjelloun Touimi Ghita ◽  
Laila Bennani ◽  
Sanae Berrada ◽  
Moussa Benboubker ◽  
Bahia Bennani

Escherichia coli is related to foodborne disease and outbreaks worldwide. It mainly affects persons at high risk as newborns, infants, and individuals with impaired immune system in hospitals. Multidrug-resistant E. coli is currently spreading both in community and hospital settings. Our study aims to evaluate the presence of E. coli and the incidence of its antibiotic resistance in samples obtained from various cooked and raw foods (N = 300), food contact surfaces (N = 238), and food handlers (N = 40) in Moroccan hospital catering service. E. coli was identified using API 20E, and the antibiotic resistance patterns were obtained using the agar disk diffusion methods. However, PCR method was used for O157 and H7 typing. The samples analysis showed that 14.33%, 24.16%, and 45% of food, surfaces, and food handlers harbored E. coli, respectively, with the highest rates obtained in raw meats (34.88%) and salads (34.88%). Molecular amplification shows that 14 E. coli isolates carried the flagellar antigen H7, while there are no isolates showing amplification for O157. The high rate of resistance was noted against ampicillin (100%), amoxicillin-clavulanate acid (100%), nalidixic acid (61.62%), and cefotaxime (59.49%), and isolates obtained from food handler’s hands showed the highest rates of resistance. None of the isolates are extended-spectrum beta-lactamases producing, while 27.7% of the isolates were metallo-beta-lactams producing. This first study conducted on Moroccan hospital catering services may draw the authorities’ attention to the necessity of setting up a surveillance system to monitor the food preparation process and the safety of prepared food in healthcare settings.


2018 ◽  
Vol 81 (8) ◽  
pp. 1339-1345 ◽  
Author(s):  
KAFEEL AHMAD ◽  
FARYAL KHATTAK ◽  
AMJAD ALI ◽  
SHAISTA RAHAT ◽  
SHAZIA NOOR ◽  
...  

ABSTRACT We report the prevalence of extended-spectrum β-lactamases and carbapenemases in Escherichia coli isolated from retail chicken in Peshawar, Pakistan. One hundred E. coli isolates were recovered from retail chicken. Antibiotic susceptibility testing was carried out against ampicillin, chloramphenicol, kanamycin, nalidixic acid, cephalothin, gentamicin, sulfamethoxazole-trimethoprim, and streptomycin. Phenotypic detection of β-lactamase production was analyzed through double disc synergy test using the antibiotics amoxicillin-clavulanate, cefotaxime, ceftazidime, cefepime, and aztreonam. Fifty multidrug-resistant isolates were screened for detection of sul1, aadA, cmlA, int, blaTEM, blaSHV, blaCTX-M, blaOXA-10, blaVIM, blaIMP, and blaNDM-1 genes. Resistance to ampicillin, nalidixic acid, kanamycin, streptomycin, cephalothin, sulfamethoxazole-trimethoprim, gentamicin, cefotaxime, ceftazidime, aztreonam, cefepime, amoxicillin-clavulanate, and chloramphenicol was 92, 91, 84, 73, 70, 67, 53, 48, 40, 39, 37, 36, and 23% respectively. Prevalence of sul1, aadA, cmlA, int, blaTEM, blaCTX-M, blaIMP, and blaNDM-1 was 78% (n = 39), 76% (n = 38), 20% (n = 10), 90% (n = 45), 74% (n = 37), 94% (n = 47), 22% (n = 11), and 4% (n = 2), respectively. blaSHV, blaOXA-10, and blaVIM were not detected. The coexistence of multiple antibiotic resistance genes in multidrug-resistant strains of E. coli is alarming. Hence, robust surveillance strategies should be developed with a focus on controlling the spread of antibiotic resistance genes via the food chain.


Antibiotics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 447
Author(s):  
Barbara Kot ◽  
Agata Grużewska ◽  
Piotr Szweda ◽  
Jolanta Wicha ◽  
Urszula Parulska

The aim of this study was to determine antibiotic resistance patterns and the prevalence of uropathogenes causing urinary tract infections (UTIs) in patients hospitalized in January–June 2020 in central Poland. Antimicrobial susceptibility testing was performed using the disk-diffusion method. Escherichia coli (52.2%), Klebsiella pneumoniae (13.7%), Enterococcus faecalis (9.3%), E. faecium (6.2%), and Proteus mirabilis (4,3%) were most commonly isolated from urine samples. E. coli was significantly more frequent in women (58.6%) (p = 0.0089) and in the age group 0–18, while K. pneumoniae was more frequent in men (24.4%) (p = 0.0119) and in individuals aged 40–60 and >60. Gram-negative species showed resistance to ampicillin. K. pneumoniae were resistant to amoxicillin plus clavulanic acid (75.0%), piperacillin plus tazobactam (76.2%), cefotaxime (76.2%), cefuroxime (81.0%), ciprofloxacin (81.0%), and trimethoprim plus sulphamethoxazole (81.0%). Carbapenems were effective against all E. coli and P. mirabilis. Some K. pneumoniae (13.6%) produced metallo-β-lactamases (MBLs). E. coli (22.6%), K. pneumoniae (81.8%), and all E. faecium were multidrug-resistant (MDR). Some E. coli (26.2%), K. pneumoniae (63.6%), and P. mirabilis (14.3%) isolates produced extended-spectrum beta-lactamases (ESBL). Vancomycin-resistant E. faecium was also found. This study showed that the possibilities of UTIs therapy using available antibiotics become limited due to the increasing number of antibiotic-resistant uropathogens.


Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1530
Author(s):  
Amanuel Balemi ◽  
Balako Gumi ◽  
Kebede Amenu ◽  
Sisay Girma ◽  
Muuz Gebru ◽  
...  

A study was carried out from August 2017 to February 2018 on lactating dairy cows, one-humped dromedary camels, and goats to determine mastitis in the Bule Hora and Dugda Dawa districts of in Southern Ethiopia. Milk samples from 564 udder quarters and udder halves from 171 animals consisting of 60 dairy cows, 51 camels, and 60 goats were tested for mastitis. Sixty-four positive udder milk samples were cultured, and bacterial mastitis pathogens were isolated and identified. The antibiotic resistance of bacterial isolates from milk with mastitis was tested against nine antimicrobials commonly used in the study area. Cow- and quarter-level prevalence of mastitis in dairy cows, camels, and goats was 33.3%, 26.3%, and 25% and 17.6%, 14.5%, and 20%, respectively. In cattle, the prevalence was significantly higher in Dugda Dawa than in Bule Hora. Major bacterial isolates were coagulase-negative Staphylococcus species (39.1%), S. aureus (17.2%), S. hyicus (14.1%), and S. intermedius and Escherichia coli (9.4% each). In camels, udder abnormality and mastitis were significantly higher in late lactation than in early lactation. Mastitis tends to increase with parity in camels. E. coli isolates were highly resistant to spectinomycin, vancomycin, and doxycycline, whereas most S. aureus isolates were multidrug-resistant. Most of the rural and periurban communities in this area consume raw milk, which indicates a high risk of infection with multidrug-resistant bacteria. We recommend a community-focused training program to improve community awareness of the need to boil milk and the risk of raw milk consumption.


2020 ◽  
Vol 17 (3) ◽  
pp. 0710
Author(s):  
Md Fazlul Karim Khan ◽  
Shah Samiur Rashid

A significant increase in the incidence of non-O157 verotoxigenic Escherichia coli (VTEC) infections have become a serious health issues, and this situation is worsening due to the dissemination of plasmid mediated multidrug-resistant microorganisms worldwide. This study aims to investigate the presence of plasmid-mediated verotoxin gene in non-O157 E. coli. Standard microbiological techniques identified a total of 137 E. coli isolates. The plasmid was detected by Perfectprep Plasmid Mini preparation kit. These isolates were subjected to disk diffusion assay, and plasmid curing with ethidium bromide treatment. The plasmid containing isolates were subjected to a polymerase chain reaction (PCR) for investigating the presence of plasmid mediated verotoxin gene (VT1 and VT2) in non-O157 E. coli. Among the 137 E. coli isolates, 49 isolates were non-O157 E. coli while 29 (59.1%) isolates were verotoxin producing non-O157 serotypes and 26 non-O157 VTEC isolates possessed plasmids. Certain isolates harboured single sized plasmid while others had multiple plasmids with different size varied from 1.8kb to 7.6kb. A plasmid containing all (100%) the isolates was multidrug-resistant. Eight isolates changed their susceptibility patterns while three isolates were found to lose plasmid after post plasmid curing treatment and the rest of the isolates (15) remained constant. Different PCR sets characterized 3 plasmid-mediated verotoxins producing non-O157 E. coli. This current study demonstrated the occurrence of plasmid mediated verotoxin gene in non-O157 E. coli. To the best of our knowledge, this is the first report in the global literature on plasmid-mediated verotoxin gene in non-O157 E. coli. Timely diagnosis and surveillance of VTEC infections should prioritize to stop or slow down the virulence gene for dissemination by plasmid-mediated gene transfer amongst the same bacteria or other species.


Author(s):  
M.A.A. Machado ◽  
W.A. Ribeiro ◽  
V.S. Toledo ◽  
G.L.P.A. Ramos ◽  
H.C. Vigoder ◽  
...  

Background: Milk is a reservoir for several groups of microorganisms, which may pose health risks. The aim of this work was to assess the antibiotic resistance and biofilm production in catalase-positive Gram-positive cocci isolated from Brazilian pasteurized milk. Methods: The bacteria were isolated using Baird-Parker agar and identified by MatrixAssisted Laser Desorption/Ionization-Time-Of-Flight (MALDI-TOF) mass spectrometer. Disk diffusion technique was used to evaluate antimicrobial susceptibility. For qualitative evaluation of biofilm production, the growth technique was used on Congo Red Agar. Results: Totally, 33 out of 64 isolates were identified, including Staphylococcus epidermidis (n=3; 4.7%), Macrococcus caseolyticus (n=14; 21.9%), and Kocuria varians (n=16; 25%). Twenty-two isolates were resistant to at least one antibiotic. Biofilm production was detected in only 5 isolates of K. varians and 1 isolate of S. epidermidis. All 14 M. caseolyticus isolates were resistant to at least one antibiotic; but, multidrug resistant (MDR) isolates were not detected. Among all K. varians isolates, 4 were resistant to at least one antibiotic from three different classes and were considered to be MDR. Conclusion: The presence of antibiotic-resistant M. caseolyticus, S. epidermidis, and K. varians isolates, especially MDRs, in milk samples highlights the possible role of milk as a reservoir of resistance genes


2021 ◽  
Author(s):  
Golnaz Mobasseri ◽  
Thong Kwai Lin ◽  
Cindy Shuan Ju Teh

Abstract Multidrug-resistant (MDR) Klebsiella pneumoniae (K. pneumoniae) poses a serious public health threat. K. pneumoniae strains that produce extended-spectrum beta-lactamases (ESBL) are becoming increasingly reported in nosocomial and community-acquired infections. Besides resistance genes, integrons, and plasmids, altered membrane permeability caused by porin loss and energy-dependent efflux have also contributed to antibiotic resistance in K. pneumoniae. The objective of this study was to determine the correlation between the reduction of antibiotic susceptibility and overexpression of efflux pump as well as the lack of outer membrane proteins (OMPs) among clinical ESBLs resistant K. pneumoniae. The expression levels of ramA, acrA, ompK35 and ompK36 in 12 MDR K. pneumoniae strains with varying MICs levels were analyzed using quantitative real time-Polymerase Chain Reaction (qRT-PCR). The role of efflux pump on antibiotic resistance was also studied by using minimum inhibitory concentration (MICs) method with//without efflux pump inhibitor. The result indicated that the strains with highest resistance to cefotaxime showed the lowest level of ompK35 and ompK36 genes expression while the strains with lowest MIC level of resistance to cefotaxime showed the highest level of expression of acrA and ramA. Our finding also revealed the effect of efflux pump inhibitor phenyl-arginine-b-naphthylamide (PAβN) on the MIC levels of ceftazidime, amoxicillin-clavulanate and cefotaxime which were significantly reduced around 1–7 folds MIC levels. These results suggest that Efflux pump system and deficiently of OMPs contributing role in antibiotic susceptibility which should be taken seriously to prevent the treatment failure due to antimicrobial resistance.


Author(s):  
Yuji Watanabe ◽  
Masafumi Seki

Antimicrobial stewardship team (AST) and Infection Control Team (ICT) have recently been linked Infectious diseases (ID) physicians, and implemented in clinical settings in Japan. The microbiological effects of an AST and ICT, in addition to Diagnostic stewardship team (DST) supported by ID physicians in our tertiary hospital were shown in significant reduction of antibiotic resistance of Escherichia coli (E coli) including extended spectrum beta-lactamases (ESBL)-producing E coli.


2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Sanjay Mahato ◽  
Ajay Mahato ◽  
Elina Pokharel ◽  
Ankita Tamrakar

Abstract Objective This study was aimed to determine prevalence and resistance pattern like multidrug resistant (MDR) or ESBL nature of E. coli and Klebsiella spp. from various sewage drain samples with an idea to deliver baseline information that could be utilized for defining guidelines for the treatment of hospital sewages. Results Of 10 sewage samples analyzed, 7 (70%) contained E. coli while 6 (60%) contained Klebsiella. Except one sample, all positive samples contained both E. coli and Klebsiella spp. E. coli isolates were resistant to ampicillin, amoxicillin, cefoxitin, cefuroxime, and cefpodoxime; while 85.7% were resistant to amoxicillin/clavulanate, ceftazidime, cefotaxime and ceftriaxone. 71.4%, 57.1%, 42.9%, and 28.6% were resistant to aztreonam, trimethoprim/sulfamethoxazole, nitrofurantoin, and gentamicin. Most were sensitive to chloramphenicol, ofloxacin, ciprofloxacin, and azithromycin. 85.7% and 57.1% of E. coli were MDR and ESBL isolates, respectively. Klebsiella were resistant to ampicillin, amoxicillin, and amoxicillin/clavulanate. 83.4% of Klebsiella were resistant to cefoxitin. 66.7% of strains were resistant to cefuroxime, ceftazidime, cefotaxime, ceftriaxone, and cefpodoxime. Klebsiella showed 50% resistant to aztreonam and trimethoprim/sulfamethoxazole, while 33.3% were resistant to chloramphenicol, nitrofurantoin, ofloxacin, and ciprofloxacin. Klebsiella were sensitive to azithromycin and gentamicin. 66.7% and 33.3% of Klebsiella were MDR and ESBL isolates, respectively.


2020 ◽  
Vol 14 (3) ◽  
pp. 126-135
Author(s):  
Mario Mitra ◽  
Andrea Mancuso ◽  
Flavia Politi ◽  
Alberto Maringhini

Bacterial infections are frequent complications of liver cirrhosis, accounting for severe clinical courses, and increased mortality. The reduction of the negative clinical impact of infections may be achieved by a combination of prophylactic measures to reduce the occurrence, early identification, and management. Spontaneous bacterial peritonitis (SBP), urinary tract infections, pneumonia, cellulitis, and spontaneous bacteremia are frequent in cirrhosis. The choice of initial empirical antimicrobial therapy should be based on both site, severity, and origin of infection (community-acquired, nosocomial, or healthcare-associated) and on antibiotic resistance patterns. 3rd generation cephalosporins are generally indicated as empirical therapy in most community-acquired cases. However, for nosocomial and healthcare-associated infections, due to a high rate of multidrug-resistant (MDR) pathogens, a broader spectrum treatment is appropriate. In order to prevent antibiotic resistance emergence, microbiological cultures should be collected, and a de-escalation applied when antimicrobial susceptibility tests are available. Standard measures to prevent infections and the identification of carriers of MDR bacteria are essential strategies to prevent infections in cirrhosis. Antibiotic prophylaxis should be applied only to gastrointestinal bleeding, SBP recurrence prevention, and cirrhotics at high risk of a first episode of SBP.


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