Diarrhoea continues to be one of the most common causes of morbidity and mortality among infants and children in developing countries. Diarrhoeagenic Escherichia coli (DEC) is an emerging agent among pathogens that cause diarrhoea. Between March 2011 and January 2012, a total of 600 stool specimens from children younger than 5 years of age (450 with and 150 without diarrhoea) were investigated for enteroaggregative E. coli (EAEC), enterohaemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC) and enterotoxigenic E. coli (ETEC) using PCR. Antimicrobial susceptibility testing was performed following Clinical and Laboratory Standards Institute guidelines. The prevalence of DEC pathotypes was 30.4 % (137 patients) and 12 % (18 patients) in the diarrhoea group and the control group, respectively. The most frequently isolated pathotype in diarrhoeal children was ETEC. This pathotype was detected significantly more often in children with diarrhoea (14.4 %) than in children without diarrhoea (5.3 %). EAEC and EPEC were detected with slightly higher frequencies in children with (8 and 4.2 %, respectively) than in children without (4.6 and 2 %, respectively) (P.0.05) diarrhoea. EHEC was only detected in children with diarrhoea (3.8 %). Of the children from the diarrhoea group, 10 % were colonized with more than one DEC pathotype. The DEC isolates exhibited high-level resistance to erythromycin (100 %), azteronam (80.7 %), amoxicillin (74.4 %) and tetracycline (69.3 %), and 86.4 % of isolates were multidrug resistant. In conclusion, ETEC continues to be an important agent associated with diarrhoea in children from Tabriz, Iran.
Background and aim
Methicillin-resistant
Staphylococcus aureus
(MRSA) is one of the most common causes of surgical infection, and its resistance to numerous conventional antibiotics makes treatment difficult. Although vancomycin is often an effective agent for the initial therapy of MRSA, clinical failure sometimes occurs. Therefore, there is an urgent need to develop better therapies. Here, we prepared some vancomycin-loaded nanoliposomes coupled with anti-staphylococcal protein (lysostaphin) and evaluated their in vitro and in vivo efficacy as a topical MRSA therapy.
Methods
Vancomycin was encapsulated in liposomes, and the coupling of lysostaphin with the surface of liposomes was carried out through cyanuric functional groups. The bactericidal efficacies and a full characterization were evaluated. To define different nanoliposomal–bacterium interactions and their bactericidal effect, flow cytometry was employed. Finally, in vivo, the topical antibacterial activity of each formulation was measured against surgical wound MRSA infection in a mouse model.
Results
High encapsulation and conjugation efficiency were achieved for all formulations. All the formulations showed a significant reduction in bacterial counts (
p<
0.05). The targeted liposomes more effectively suppress bacterial infection in vitro and in vivo relative to equivalent doses of untargeted vancomycin liposome. The flow cytometry results confirmed liposome–bacterium interactions, which increased during the incubation time. The maximum binding rate and the bactericidal effect were significantly higher in targeted liposomes (
p<
0.05) compared with control liposomes.
Conclusion
Our data suggest a novel nano-vehicle (lysostaphin-conjugated coupled liposomal vancomycin) which could be used as a great topical antimicrobial construct for treatment of MRSA skin infections.
Integrons are considered to play a significant role in the evolution and spread of antibiotic resistance genes. A total of 349 clinical isolates of Escherichia coli and Klebsiella pneumoniae were investigated for molecular characterization of integrons and betalactamases. Antimicrobial susceptibility testing was also performed as the Clinical and Laboratory Standards Institute (CLSI) guidelines. The frequency of extended spectrum betalactamases (ESBL) or metallo-betalactamases (MBL)-producing isolates, patient demographics, and the susceptibility to various antimicrobial agents were described. BlaCTX-M was the most frequently detected betalactamase in all isolates. Moreover, MBL producing K. pneumoniae carried blaIMP and blaVIM at 100 and 41·6%, respectively but no MBL-positive E. coli was detected. Class 1 integrons were more frequent among E. coli and K. pneumoniae isolates in comparison with class 2 integrons and the frequency of intI2 in K. pneumoniae was significantly higher than E. coli isolates. Five different resistance gene arrays were identified among class 1 integrons. Dihydrofolate reductase (dfrA) and aminoglycoside adenyltransferase (aad) gene cassettes were found to be predominant in the class 1 integrons. These results indicate that class 1 integrons are widespread among ESBL-producing isolates of K. pneumoniae and E. coli and appropriate surveillance and control measures are essential to prevent further dissemination of these elements among Enterobacteriaceae in our country.
Multi-drug-resistant (MDR) diarrheagenic Escherichia coli (DEC) has rapidly spread worldwide and represents the most serious threat to the management of diarrhea in developing countries. During the period from March 2011 to January 2012, a total of 450 stool samples of diarrheal children aged 0-60 months were studied. In order to detect enterotoxigenic E. coli (ETEC) and enterohemorrhagic E. coli (EHEC) simultaneously, a mixture of four primer pairs specific for eltB, estA, vt1, and vt2 genes was used in a multiplex PCR. Antimicrobial susceptibility testing was performed as the Clinical and Laboratory Standards Institute (CLSI) guidelines. A total of 140 (31·1%) DEC were isolated from 450 stool samples. Diarrheagenic E. coli exhibited high-level resistance to aztreonam (80·7%), amoxicillin (74·4%), and tetracycline (69·3%). Also, 86·4% of E. coli isolates were resistant to at least three different classes of antimicrobial agents and considered as MDR. The frequency of ETEC and EHEC pathotypes was 46·4 and 12·1%, respectively and all of these isolates were MDR. In conclusion, MDR ETEC continues to be an important agent associated with diarrhea in children from Tabriz, Iran.
ObjectivesIntegrons are thought to play an important role in the spread of antibiotic resistance. This study investigates class 1 and 2 integron-positive methicillin-resistant coagulase-negative staphylococci strains isolated in Iran and characterizes their patterns of antimicrobial resistance.MethodsHundred clinical isolates of coagulase-negative staphylococci were characterized for integron content and staphylococcal cassette chromosome mec (SCCmec) type.ResultsSixteen isolates carried class 1 (intI1) integrons and four isolates carried class 2 (intI2) integrons. One resistance gene array was identified among the class 1 integrons (aadA1 cassette). The distribution of SCCmec types in 50 methicillin-resistant coagulase-negative staphylococci strains showed that SCCmec types III and V dominated among the tested strains.ConclusionThis is the first report of methicillin-resistant coagulase-negative staphylococci strains that carry two mobile genetic elements, including class 1 and 2 integrons and SCCmec, in Iran.
Background: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common causes of skin infections and treatment is difficult due to its resistance to the most of antibiotics. Although vancomycin is often considered as an antibacterial agent of choice for the treatment of MRSA, its use is limited because of the high side effects. One solution is using liposomal formulation for local drug delivery. The aim of this study was to determine in vitro and in vivo efficacies of liposomal vancomycin as topical use. Methods: To prepare liposomal vancomycin, the ammonium sulfate gradient using remote loading and freeze-thaw methods was applied. Then, synthesized nanoliposomes were evaluated in terms of particle size, morphology, stability, and encapsulation efficiency. Minimum inhibitory concentration (MIC) of synthesized nanoliposome against MRSA was detected. The cytotoxicity of synthesized nanoliposome was evaluated using MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Finally, the topical antibacterial activity of each formulation was tested against MRSA-infected skin wound model in mice. Results: High encapsulation efficiency was achieved for all synthesized nanoliposomes. The results of in vitro and in vivo showed that liposomal vancomycin was more effective than free vancomycin. Also, synthesized nanoliposome showed no cytotoxicity on human epidermoid cell line. Conclusion: The results showed that synthesized nanoliposome could be applied as a great topical antimicrobial construct for treatment of MRSA skin infections.
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