Recurrent urinary tract infection with antibiotic-resistant Klebsiella pneumoniae in a patient with Crohn's disease: A case report.

Recurrent urinary tract infections with resistant strains of Klebsiella pneumoniae are a potential complication of the long-term use of immunosuppressive therapy in patients with Crohn's disease.

A Recombinant Snake Cathelicidin Derivative Peptide: Antibiofilm Properties and Expression in Escherichia coli.

The emergence of antimicrobial resistance among pathogenic microorganisms has been led to an urgent need for antibiotic alternatives. Antimicrobial peptides (AMPs) have been introduced as promising therapeutic agents because of their remarkable potentials. A new modified cathelicidin-BF peptide (Cath-A) with 34 amino acid sequences, represents the potential antimicrobial effects against methicillin-resistant Staphylococcus aureus (MRSA) with slight hemolytic and cytotoxic activities on eukaryotic cells. In this study, the effects of Cath-A on Acinetobacter baumannii, and Pseudomonas aeruginosa isolated from medical instruments were studied. Cath-A inhibited the growth of bacterial cells in the range of 8⁻16 µg/mL and 16-≥256 µg/mL for A. baumannii and P. aeruginosa, respectively. The peptide significantly removed the established biofilms. To display a representative approach for the cost-effective constructions of peptides, the recombinant Cath-A was cloned in the expression vector pET-32a(+) and transformed to Escherichia coli BL21. The peptide was expressed with a thioredoxin (Trx) sequence in optimum conditions. The recombinant peptide was purified with a Ni2+ affinity chromatography and the mature peptide was released after removing the Trx fusion protein with enterokinase. The final concentration of the partially purified peptide was 17.6 mg/L of a bacterial culture which exhibited antimicrobial activities. The current expression and purification method displayed a fast and effective system to finally produce active Cath-A for further in-vitro study usage.

Carvacrol and human health: A comprehensive review.

Carvacrol (CV) is a phenolic monoterpenoid found in essential oils of oregano (Origanum vulgare), thyme (Thymus vulgaris), pepperwort (Lepidium flavum), wild bergamot (Citrus aurantium bergamia), and other plants. Carvacrol possesses a wide range of bioactivities putatively useful for clinical applications such antimicrobial, antioxidant, and anticancer activities. Carvacrol antimicrobial activity is higher than that of other volatile compounds present in essential oils due to the presence of the free hydroxyl group, hydrophobicity, and the phenol moiety. The present review illustrates the state-of-the-art studies on the antimicrobial, antioxidant, and anticancer properties of CV. It is particularly effective against food-borne pathogens, including Escherichia coli, Salmonella, and Bacillus cereus. Carvacrol has high antioxidant activity and has been successfully used, mainly associated with thymol, as dietary phytoadditive to improve animal antioxidant status. The anticancer properties of CV have been reported in preclinical models of breast, liver, and lung carcinomas, acting on proapoptotic processes. Besides the interesting properties of CV and the toxicological profile becoming definite, to date, human trials on CV are still lacking, and this largely impedes any conclusions of clinical relevance.

The antimicrobial potential of a new derivative of cathelicidin from Bungarus fasciatus against methicillin-resistant Staphylococcus aureus.

Cathelicidins are a family of antimicrobial peptides which exhibit broad antimicrobial activities against antibiotic-resistant bacteria. Considering the progressive antibiotic resistance, cathelicidin is a candidate for use as an alternative approach to treat and overcome the challenge of antimicrobial resistance. Cathelicidin-BF (Cath-BF) is a short antimicrobial peptide, which was originally extracted from the venom of Bungarus fasciatus. Recent studies have reported that Cath-BF and some related derivatives exert strong antimicrobial and weak hemolytic properties. This study investigates the bactericidal and cytotoxic effects of Cath-BF and its analogs (Cath-A and Cath-B). Cath-A and Cath-B were designed to increase their net positive charge, to have more activity against methicillin resistant S. aureus (MRSA). The results of this study show that Cath-A, with a +17-net charge, has the most noteworthy antimicrobial activity against MRSA strains, with minimum inhibitory concentration (MIC) ranging between 32-128 µg/ml. The bacterial kinetic analysis by 1 × MIC concentration of each peptide shows that Cath-A neutralizes the clinical MRSA isolate for 60 min. The present data support the notion that increasing the positive net charge of antimicrobial peptides can increase their potential antimicrobial activity. Cath-A also displayed the weakest cytotoxicity effect against human umbilical vein endothelial and H9c2 rat cardiomyoblast cell lines. Analysis of the hemolytic activity reveals that all three peptides exhibit minor hemolytic activity against human erythrocytes at concentrations up to 250 µg/ml. Altogether, these results suggest that Cath-A and Cath-B are competent candidates as novel antimicrobial compounds against MRSA and possibly other multidrug resistant bacteria.

Emergence of blaCTX-M-15, blaTEM-169 and blaPER-1 extended-spectrum ß-lactamase genes among different Salmonella enterica serovars from human faecal samples.

BACKGROUND: Broad-spectrum ß-lactams are used for empirical therapy of severe infections with non-typhoid Salmonella serotypes; however, activities of these drugs against the strains producing different ß-lactamase is not so clear. This study investigated the prevalence of ß-lactamase genes among isolates of S. enterica serovars from human faecal samples and determined their diversity in activity against different ß-lactams. METHODS: Antimicrobial resistance of faecal isolates of S. enterica to extended-spectrum cephalosporins was analysed and MIC values were determined for the strains presenting extended-spectrum ß-lactamases (ESBLs) phenotypes. The ß-lactamase genes were identified by PCR and sequencing. ß-lactamase activity of the Salmonella strains exhibiting ESBL phenotype was detected by biological, iodometric, spectrophotometry and nitrocefin assays. RESULTS: Out of 202 S. enterica isolates, ESBLs phenotype was detected among 3.4% (7/202) of the strains. blaTEM-1 and blaCTX-M-15 were among the frequent ß-lactamase genes. Detection of blaTEM-169 in S. enterica serovar Typhimurium and S. enterica serovar Bredeney and blaPER-1 in S. enterica serovar Infantis was a new finding in this experiment. Location of blaCTX-M-15/blaTEM-169/blaPER-1 genes on plasmid was confirmed in a transformation experiment. While crude extracts of the enzymes from each strain showed higher activity against cephalothin and cefotaxime, the lowest activity was detected against ceftazidime. The greatest synergistic activity was seen in a strain of S. enterica that carried blaCTX-M-15 and blaPER-1 genes compared with those presenting blaCTX-M-15/blaTEM-169 or blaCTX-M-15/blaTEM-1 genotypes. CONCLUSIONS: The results show dissemination of ESBLs encoding genes and their combined activity among different serovars of S. enterica that are a threat for future treatment options.

Antimicrobial-resistant Shigella infections from Iran: an overlooked problem?

OBJECTIVES: In this study, we wanted to assess the level of antimicrobial resistance, the presence of genes encoding resistance to cephalosporins and plasmid-mediated quinolone resistance (PMQR), and genetic relatedness among Shigella isolates obtained from Iranian patients. METHODS: A total of 44 Shigella isolates were collected from Iranian patients admitted to Milad Hospital, Tehran, Iran, during 2008-10. Of these, 37 were serotyped and characterized by MIC determination. A subset of eight suspected extended-spectrum ß-lactamase (ESBL) producers (six Shigella sonnei phase II and two Shigella flexneri type 1b) were examined for the presence of genes encoding cephalosporin resistance. The presence of PMQR was assessed in one S. flexneri isolate exhibiting low-level resistance to ciprofloxacin and susceptibility to nalidixic acid. PFGE was performed on 25 S. sonnei phase II isolates. RESULTS: Of the isolates, 25 (68%) were S. sonnei phase II, with 5 (14%) S. flexneri, 5 (14%) Shigella dysenteriae type 2, and 2 (5%) Shigella boydii type 2. Resistance to at least threeclasses of antimicrobials was detected in all species. The presence of bla(CTX-M-15) and the AmpC ß-lactamase producer bla(CMY-2) was confirmed in five and one S. sonnei phase II isolates, respectively. One of the two S. flexneri type 1b that contained bla(CTX-M-15) also harboured a qnrS1 gene. PFGE identified sevenPFGE profiles; the main cluster included 15 of the strains, suggesting low genetic diversity between isolates or the presence of an endemic clone in Iran. CONCLUSIONS: This is the first known description of ESBL-producing and AmpC ß-lactamase-producing Shigella and of PMQR Shigella in Iran. The emergence of CTX-15, CMY-2 and qnrS1 genes may compromise the treatment of shigellosis. Strategies to minimize the spread of ESBL-producing and AmpC-ß-lactamase-producing Shigella should be implemented.

Antimicrobial resistance in Salmonella spp. recovered from patients admitted to six different hospitals in Tehran, Iran from 2007 to 2008.

The objective of this study was to assess the genotypic diversity associated with antimicrobial susceptibility of Salmonella serovars isolated from patients arriving with diarrhoea to six hospitals of Tehran, Iran. During 2007-2008, a cross-sectional convenience study was performed. Stool samples were screened for the presence of Salmonella, serotyped, tested for antimicrobial susceptibility using disk diffusion and examined for the presence of relevant resistance genes and integrons by PCR. A total of 1,120 patients were screened for the presence of Salmonella. Out of 71 Salmonella isolates recovered, the following serovars were identified: 17 Typhi, 14 Paratyphi C, 13 Enteritidis, 11 Paratyphi B, 10 Paratyphi A and six Infantis. Most resistance was observed towards sulfamethoxazole (30%), tetracyclines (25%), nalidixic acid (22%), spectinomycin (17%), trimethoprim (15%), ampicillin (14%) and kanamycin (14%). The tetracycline resistance genes tet(A), tet(B), and tet(G) were found in 28%, 14% and 6% of the tetracycline resistant isolates, respectively. The genes aadA, aadB, strA, strB and aphA1-Iab were present in 83%, 55%, 34%, 1% and 17% of the aminoglycoside resistant isolates, respectively. Additionally, bla (PSE) and bla (TEM) ß-lactamase genes were detected in 63% and 18% of the ampicillin-resistant isolates. The 23 sulphonamide resistant isolates harboured sul1 and intI1 genes, typical to class 1 integrons. Nine of these isolates also yielded amplicons for intI2 (class 2 integrons). The presence of multi-drug resistant Salmonella may compromise the successful treatment of enteric infection diseases. The enforcement of strict prescription practices will help to minimise the emergence of resistance.

Phylogenetic relationship of Salmonella enterica strains in Tehran, Iran, using 16S rRNA and gyrB gene sequences.

INTRODUCTION: We assessed whether 16S rDNA and gyrB gene sequences, alone or combined, were suitable for determining the phylogenetic relationship among Salmonella enterica strains isolated from Tehran, Iran. Patients over five years of age enrolled in an acute diarrheal surveillance project in Tehran province between May 2004 and October 2006 were selected as our study group. METHODOLOGY: 16S ribosomal DNA (rDNA) and gyrB genes from 40 Salmonella isolates obtained from patients with acute diarrhea were sequenced and the data was used to generate phylogenetic trees that facilitated isolate comparison. RESULTS: Salmonella strains clustered into five to seven phylogenetic groups, dependent on analysis of 16S rDNA (1546 bp), gyrB (1256 bp) or a combination of the two genes. By 16S rDNA sequence analysis, only strains of Salmonella enterica serovar Typhi ( S. Typhi) clustered exclusively together. gyrB sequences permitted clustering of all the S. Typhi and S. Paratyphi A isolates, and clustering of S. Enteritidis into two separate but exclusive groups. Concatenation of the two data sets did not significantly improve the resolution of the strains compared to the gyrB gene. None of the analyses completely resolved S. enterica Paratyphi B and C into mutually exclusive groups. CONCLUSION: Sequencing of gyrB represents a potentially useful tool for determining the phylogenetic relationship of S. enterica strains in Tehran, Iran. Genetic analysis of the 16S rRNA gene alone or in combination with gyrB did not increase the resolution between serotypes of S. enterica. We speculate that inclusion of additional genetic markers would improve the sensitivity of the analysis.

Detection of novel gyrA mutations in nalidixic acid-resistant isolates of Salmonella enterica from patients with diarrhoea.

The aim of the current study was to detect mutations in the gyrA gene of quinolone-resistant Salmonella spp. isolates recovered in Tehran, Iran. Between April 2008 and September 2009, 174 Salmonella spp. were collected and assayed for quinolone resistance and detection of gyrA mutations. Isolates identified as Salmonella enterica were tested for susceptibility by the disk diffusion method. Polymerase chain reaction (PCR) amplification and sequencing of the gyrA gene segment encoding the quinolone resistance-determining region (QRDR) were performed for the nalidixic acid-resistant isolates. Amongst the 174 recovered Salmonella spp. isolates, 89 were resistant to nalidixic acid, of which 9 were resistant to enrofloxacin; 10 isolates had reduced susceptibility to nalidixic acid. None of the isolates were resistant to ciprofloxacin, but a single isolate showed reduced susceptibility. Twelve types of amino acid replacement were found in the QRDR region of GyrA, namely the previously described substitutions in positions 83 and 87 as well as five new substitutions Leu41-Pro, Arg47-Ser, Ser111-Thr, Ala118-Thr and Asp147-Gly. Double substitutions in both positions 83 and 87 were not identified. A Gly133-Glu substitution was identified in a single S. enterica serotype Typhi isolate.

Emergence of extended-spectrum beta-lactamases in clinical isolates of Salmonella enterica in Tehran, Iran.

The purpose of the current study was to investigate the presence and molecular type(s) of extended-spectrum beta-lactamases (ESBLs) in Salmonella spp. isolates obtained from patients with diarrhea in hospitals of Tehran, Iran. Over a period of 17 months, 129 Salmonella spp. were isolated from fecal samples and tested for susceptibility using the Kirby-Bauer disk diffusion method; then, screening for ESBL-producing isolates and determination of their minimum inhibitory concentrations were carried out using the combined disk method and standard agar dilution method, respectively. The presence and type of ESBL-encoding genes were determined by PCR and sequence analysis. The isolates were all identified as Salmonella enterica of different serovars. The highest resistance in the collected Salmonella isolates was to nalidixic acid (45.7%), followed by tetracycline (43.4%), trimethoprim-sulfamethoxazole (36.4%), ampicillin (15.5%), and chloramphenicol (14.7%). All the isolates were susceptible to ciprofloxacin, gentamicin, and cefoxitin. Three S. enterica isolates were resistant to ampicillin, piperacillin, ceftazidime, cefotaxime, ceftriaxone, cefpodoxime, cephalothin, and aztreonam. PCR and DNA sequencing revealed that two of the three isolates harbored both a bla(CTX-M-15) and a bla(TEM) gene while the third one carried only a bla(CTX-M-15) gene. This is the first study providing structural data for a CTX-M-type beta-lactamase produced by Salmonella isolates recovered in Iran.