High prevalence of multidrug-resistant Enterobacterales carrying extended-spectrum beta-lactamase and AmpC genes isolated from neonatal sepsis in Ahvaz, Iran.

OBJECTIVES: In the recent years, multidrug resistant (MDR) neonatal septicemia-causing Enterobacterales has been dramatically increased due to the extended-spectrum beta-lactamases (ESBLs) and AmpC enzymes. This study aimed to assess the antibiotic resistance pattern, prevalence of ESBLs/AmpC beta-lactamase genes, and Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) fingerprints in Enterobacterales isolated from neonatal sepsis. RESULTS: In total, 59 Enterobacterales isolates including 41 (69.5%) Enterobacter species, 15 (25.4%) Klebsiella pneumoniae and 3 (5.1%) Escherichia coli were isolated respectively. Resistance to ceftazidime and cefotaxime was seen in all of isolates. Furthermore, all of them were multidrug-resistant (resistant to three different antibiotic categories). The phenotypic tests showed that 100% of isolates were ESBL-positive. Moreover, AmpC production was observed in 84.7% (n = 50/59) of isolates. Among 59 ESBL-positive isolates, the highest percentage belonged to blaCTX-M-15 gene (66.1%) followed by blaCTX-M (45.8%), blaCTX-M-14 (30.5%), blaSHV (28.8%), and blaTEM (13.6%). The frequency of blaDHA, blaEBC, blaMOX and blaCIT genes were 24%, 24%, 4%, and 2% respectively. ERIC-PCR analysis revealed that Enterobacterales isolates were genetically diverse. The remarkable prevalence of MDR Enterobacterales isolates carrying ESBL and AmpC beta-lactamase genes emphasizes that efficient surveillance measures are essential to avoid the more expansion of drug resistance amongst isolates.

Molecular evaluation of aminoglycosides resistance and biofilm formation in Klebsiella pneumoniae clinical isolates: A cross-sectional study.

Background and Aims: Resistance to antibiotics and the capability to develop biofilm as two main virulent determinants of Klebsiella pneumoniae have important role in infection persistence. The aim of the study was to evaluate the association between the prevalence of aminoglycoside resistance and virulence genes and biofilm formation capacity in K. pneumoniae strains isolated from hospitalized patients in South-West of Iran. Methods: A total of 114 non-duplicate clinical isolates of K. pneumoniae collected from Ahvaz teaching hospitals. Identification of species was performed by biochemical tests and then confirmed by polymerase chain reaction (PCR) of rpoB gene. The susceptibility to antibiotics was determined by Kirby-Bauer disk diffusion method. Biofilm formation was assessed by microtiter plate method. Finally, PCR was conducted to detect virulence gene determinants including fimbrial genes, aminoglycoside modifying enzymes- and 16S rRNA methylase (RMTase) genes. Results: Totally, all collected strains were carbapenem resistant and showed multidrug- and extensively drug-resistance phenotype (75% and 25%, respectively). Seventy-one percent (n = 81) of isolates were non-susceptible to aminoglycosides. Among aminoglycoside antibiotics, K. pneumoniae isolates showed the highest and lowest resistance rates to tobramycin (71%) and the amikacin (25%), respectively. All biofilm producer strains were positive for the presence virulence determinants including ecpA, fimA, mrkD, and mrkA. Of 81 aminoglycosides non-susceptible isolates 33% were positive for the presence ant (2″)-Ia as the most prevalent gene followed by aac (3')-IIa and armA (27%), aac (6')-Ib (18%), and aph (3')-Ia (15%). Conclusion: K. pneumoniae isolates showed the highest and the lowest aminoglycoside resistance rates to tobramycin and amikacin, respectively. Majority of isolates were biofilm producers and there was significant association between antibiotic resistance pattern and the strength of biofilm production. The ant(2″)-Ia, aac (3')-IIa, and armA genes in aminoglycoside-resistant isolates.

Occurrence of diarrheagenic Escherichia coli pathotypes from raw milk and unpasteurized buttermilk by culture and multiplex polymerase chain reaction in southwest Iran.

BACKGROUND: In developing countries including Iran, there are limited data on diarrheagenic Escherichia coli (DEC) contamination in milk and unpasteurized buttermilks. This study aimed to determine the occurrence of DEC pathotypes by culture and multiplex polymerase chain reaction (M-PCR) in some dairy products from southwest Iran. METHODS AND RESULTS: In this cross-sectional study (September to October 2021), 197 samples (87 unpasteurized buttermilk and 110 raw cow milk) were collected from dairy stores of Ahvaz, southwest Iran. The presumptive E. coli isolates were primarily identified using biochemical tests and then confirmed by PCR of uidA gene. The occurrence of 5 DEC pathotypes: enterotoxigenic E. coli (ETEC), enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), and enteroinvasive E. coli (EIEC) were investigated using M-PCR. Overall, 76 (76/197, 38.6%) presumptive E. coli isolates were identified by biochemical tests. Using uidA gene, only 50 isolates (50/76, 65.8%) were confirmed as E. coli. DEC pathotypes were detected in 27 of 50 (54.0%) E. coli isolates (74.1%, 20/27 from raw cow milk and 25.9%, 7/27 from unpasteurized buttermilk). The frequency of DEC pathotypes was as follows: 1 (3.7%) EAEC, 2 (7.4%) EHEC, 4 (14.8%) EPEC, 6 (22.2%) ETEC, and 14 (51.9%) EIEC. However, 23 (46.0%) E. coli isolates had only the uidA gene and were not considered DEC pathotypes. CONCLUSION: Possible health risks for Iranian consumers can be attributed to the presence of DEC pathotypes in dairy products. Hence, serious control and prevention efforts are needed to stop the spread of these pathogens.

Identification of a New Compound (4-Fluoro-2-Trifluoromethyl Imidazole) Extracted from a New Halophilic Bacillus aquimaris Strain Persiangulf TA2 Isolated from the Northern Persian Gulf with Broad-Spectrum Antimicrobial Effect.

Background: The unique ecosystem of the Persian Gulf has made it a rich source of natural antimicrobial compounds produced by various microorganisms, especially bacteria, which can be used in the treatment of infectious diseases, especially those of drug-resistant microbes. Objectives: This study aimed to identify antimicrobial compounds in the bacteria isolated from the northern region of the Persian Gulf in Abadan (Chavibdeh port), Iran, for the first time. Materials and Methods: Sampling was performed in the fall on November 15, 2019, from 10 different stations (water and sediment samples). The secondary metabolites of all isolates were extracted, and their antimicrobial effects were investigated. 16S ribosomal ribonucleic acid sequencing was used for the identification of the strains that showed the best inhibition against selected pathogens, and growth conditions were optimized for them. A fermentation medium in a volume of 5000 mL was prepared to produce the antimicrobial compound by the superior strain. The extracted antimicrobial compounds were identified using the gas chromatography-mass spectrometry technique. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for the superior strain. The effects of salinity, pH, and temperature on the production of antimicrobial compounds were determined by measuring the inhibitory region (mm) of methicillin-resistant Staphylococcus aureus (MRSA). Results: Four new strains with antimicrobial properties (i.e., Halomonas sp. strain Persiangulf TA1, Bacillus aquimaris strain Persiangulf TA2, Salinicoccus roseus strain Persiangulf TA4, and Exiguobacterium profundum strain Persiangulf TA9) were identified. The optimum growth temperatures were determined at 37-30, 37, and 40 °C for TA1 and TA2, TA4, and TA9 strains, respectively. The optimum pH values for the four strains were 7, 6-7, 7.5, and 6.5-7.5, respectively. The optimal salt concentrations for the four strains were 15%, 2.5-5%, 7.5%, and 5%, respectively. The ethyl acetate extract of strain Persiangulf TA2 showed extensive antimicrobial activity against human pathogens (75%) and MRSA. The most abundant compound identified in TA2 extract was the new compound 4-fluoro-2-trifluoromethyl imidazole. The MBC and MIC for the ethyl acetate extract of strain TA2 were 20 and 5 mg. mL-1 (Staphylococcus aureus), 40 and 20 mg. mL-1 (MRSA, Escherichia coli, and Enterococcus faecalis), 40 and 10 mg. mL-1 Acinetobacter baumannii), and 80 and 40 mg. mL-1 (Staphylococcus epidermidis, Shigella sp., Bacillus cereus, and Klebsiella pneumoniae), respectively. The optimal conditions for antibiotic production by TA2 strain were 5% salt concentration, pH of 7, and temperature of 35 °C. Conclusion: Newly detected natural compounds in TA2 strain due to superior antimicrobial activity even against MRSA strain can be clinically valuable in pharmacy and treatment.

Loop mediated isothermal amplification of Clostridioides difficile isolates in gastrointestinal patients.

This study investigated the prevalence of Clostridioides difficile by culture, multiplex polymerase chain reaction (M-PCR), and loop mediated isothermal amplification (LAMP) in patients with suspected C. difficile infections (CDIs). Also, the results of three methods were compared. All stool specimens collected from CDI suspected patients were cultured on selective C. difficile cycloserine-cefoxitin fructose agar and incubated in an anaerobic jar up to 7 days. The bacterial isolates were identified using standard tests. Multiplex-PCR (M-PCR) was performed for detection of tcdA, tcdB, and tpi genes. The LAMP assay was performed to detect the tcdB gene of C. difficile. C. difficile was isolated from 20.0% (n = 10/50) of samples by culture. M-PCR showed that 34.0% (n = 17/50) of the specimens were positive for C. difficile based on the presence of tpi gene. Out of the 17 C. difficile, 13 strains (76.0%) were positive for tcdB gene using M-PCR. However, the LAMP assay showed that 30.0% (15/50) of specimens were positive for the presence of tcdB gene. M-PCR and LAMP methods showed 100.0% sensitivity compared to the culture method. However, the specificity of the LAMP (87.5%) was relatively higher than the M-PCR (82.5%) compared to the culture. Based on the results of this study, the prevalence of toxigenic C. difficile strains was high in suspected CDI patients. So, the differentiation between toxigenic and non-toxigenic strains is necessary. Our data showed that the LAMP assay is a good method for direct detection of toxigenic C. difficile strains from stool specimens.

Detection of OqxAB Efflux Pumps, a Multidrug-Resistant Agent in Bacterial Infection in Patients Referring to Teaching Hospitals in Ahvaz, Southwest of Iran.

Antibiotic resistance mechanisms in Enterobacteriaceae are causative agents of global health problems. Bacterial infections due to multidrug resistance (MDR) may be mediated by the overexpression of efflux pumps. In this study, we investigated the prevalence of oqxA and oqxB genes as two encoding agents of efflux pumps and the determination of antibiotic resistance rate in clinical isolates of Enterobacteriaceae. In this study, 100 Enterobacteriaceae isolates collected from different clinical specimens of infectious patients, such as wounds, urine, blood, discharge, and abscesses except stool, were examined. Identification of the isolates was performed using standard biochemical tests such as TSI, citrate, urea, lysine, SIM, MR-VP, and gas production. The antimicrobial susceptibility test was carried out by the Kirby-Bauer disk diffusion method according to CLSI guidelines, and finally, the oqxA and oqxB genes were detected by the PCR method. Among 100 Enterobacteriaceae isolates, Escherichia coli and Enterobacter gergoviae were the most common isolates with 71% and 20%, respectively. Also, the lowest isolates belonged to Enterobacter cloacae (3%) and Klebsiella pneumoniae (1%). Out of 100 Enterobacteriaceae isolates, 37 isolates (37%) were positive for at least one of oqxA or oqxB genes, while both of these genes were detected among 12% of them. oqxAB genes were detected in 8 cases of 20 (40%) Enterobacter gergoviae and 4 cases of 71 (5.7%) E. coli isolates. The antimicrobial susceptibility test showed that all isolates (100%) were susceptible to imipenem, while the maximum resistance to piperacillin, ceftriaxone, and cefotaxime were 69%, 55%, and 55%, respectively. Also, the results of this study showed that antibiotic resistance in Enterobacteriaceae isolates caused by oqxAB genes is increasing among patients in Iran. Therefore, identification of resistant isolates and antibiotic monitoring programs are essential to prevent the spread of MDR isolates.

Prevalence of methicillin resistance and superantigenic toxins in Staphylococcus aureus strains isolated from patients with cancer.

BACKGROUND: This study aimed to determine the frequency of methicillin-resistant Staphylococcus aureus (MRSA), antibiotic resistance patterns, superantigenic toxins profile, and clonality of this pathogen in patients with cancer. RESULTS: In total, 79 (25.7%) isolates were confirmed as Staphylococcus species, from which 38 (48.1%) isolates were S. aureus, and 29 (76.3%) isolates were confirmed as MRSA. The highest resistance in MRSA strains was seen against ciprofloxacin (86.2%) and erythromycin (82.8%). Teicoplanin, and linezolid were the most effective antibiotics. From all MRSA isolates, 3 strains (10.3%) were resistant to vancomycin with minimum inhibitory concentration values of 128 µg/ml. The prevalence of superantigenic toxins genes was as follows: pvl (10.5%), tsst-1 (36.8%), etA (23.7%), and etB (23.7%). The t14870 spa type with frequency of 39.5% was the most prevalent clone type circulating in the cancer patients. CONCLUSIONS: This study showed the circulating of spa t14870 as the most predominant MRSA clone in cancer patients of southwest Iran. Also, a diverse antibiotic resistance pattern and toxin profiles were seen among MRSA isolates.

Spatio-temporal variations of airborne bacteria from the municipal wastewater treatment plant: a case study in Ahvaz, Iran.

The study of bioaerosol dispersion in wastewater treatment plants (WWTPs) has received considerable attention. This study aimed to investigate the seasonal changes and spatial distributions of airborne bacteria around different parts of Ahvaz WWTP, the capital city of Khuzestan Province, Iran, over 4 months in the cold and warm seasons. Samples were collected from 2 and 10-m intervals of grit chamber (GCh), primary sludge dewatering basin (PSDB), aeration tank (AT), as well as 60-m upstream (US) and downstream (DS) of the WWTP. Further, bacteria in the indoor air of administrative building (AB) of WWTP were investigated. Bioaerosols were collected by passive sampling method. The total bacteria count was 105.3 ± 98.5 CFU/plate/h. The dominant bacteria stood 2 m away from the AT with an average 244.2 ± 73.1 CFU/plate/h in the warm season while they were the lowest with an average 43 ± 11.4 CFU/plate/h in the 10-m distance of the GCh in the cold season. According to the sequencing results, the dominant bacterial species included Bacillus pumilus (26.7%), Staphylococcus arlettae (23.2%), Kocuria turfanensis (13.6%) and Alicycliphilus (9.2%), respectively. There was a positive relationship between the release of bacteria, temperature and wind speed. However, there was a significant negative correlation between total bacteria concentration and humidity. There are accumulative perils to WWTP workers and neighbors exposed by persistent exposure to airborne bacteria. Therefore, AT should be paid more attention as a dominant source of airborne bacteria emissions, especially in the warm season.

Genotyping and molecular characterization of clinical Acinetobacter baumannii isolates from a single hospital in Southwestern Iran.

ACINETOBACTER BAUMANNII: (A. baumannii) is a pathogen responsible for nosocomial infections among the hospitalized patients. The aim of this study was to investigate genotyping and molecular characterization and to examine the biofilm formation ability of A. baumannii isolates. In total, 70 A. baumannii isolates were collected from patients admitted to Imam Khomeini Hospital in Ahvaz, Southwestern Iran. Minimum inhibitory concentrations (MIC) test was performed using Vitek 2 system. The presence of genes encoding metallo-ß-lactamases, oxacillinases, and integrase and the biofilm formation ability were then evaluated. Multiple locus variable-number tandem repeat (VNTR) analysis (MLVA) typing and multiplex PCR were performed to determine the genetic relationships. The blaOXA-23-like gene had the highest prevalence. The frequency of genes encoding blaSPM, blaIMP, and blaVIM among MDR A. baumannii isolates were 12 (17.1%), 18 (25.7%), and 22 (31.4%), respectively. Moreover, 46 isolates (75.4%) harbored class I integron and 10 isolates (16.39%) carried class II integron. The number of weak, moderate and strong biofilm-producing isolates were 3 (4.3%), 7 (10%), and 55 (78.5%), respectively. The results showed that 70 A. baumannii isolates were grouped into 12 distinct MLVA types with five clusters and four singleton genotypes. In addition, 25 (35.7%) isolates were assigned to international clone (IC) variants, 37 (52.8%) isolates belonged to group 1 (IC II), and 8  (11.4%) isolates belonged to group 2 (IC I). Our findings revealed that the population structure of the A. baumannii isolates was genetically diverse. More focus on genetic variation and antibiotic resistance of A. baumannii isolates are recommended.

Survey on Genetic Diversity, Biofilm Formation, and Detection of Colistin Resistance Genes in Clinical Isolates of Acinetobacter baumannii.

INTRODUCTION: Acinetobacter baumannii is an opportunistic pathogen responsible for nosocomial infections. The emergence of colistin-resistant A. baumannii is a significant threat to public health. The aim of this study was to investigate the molecular characterization and genotyping of clinical A. baumannii isolates in Southwestern Iran. METHODS: A total of 70 A. baumannii isolates were collected from patients admitted to Imam Khomeini Hospital in Ahvaz, Southwestern Iran. Minimum inhibitory concentration test was conducted by using Vitek 2 system. The presence of biofilm-forming genes and colistin resistance-related genes were evaluated by PCR. The isolates were also examined for their biofilm formation ability and the expression of pmrA and pmrB genes. Finally, multilocus sequence typing (MLST) and PCR-based sequence group were used to determine the genetic relationships of the isolates. RESULTS: Overall, 61 (87.1%) and 9 (12.8%) isolates were multidrug-resistant (MDR) and extensively drug-resistant (XDR), respectively. Colistin and tigecycline with 2 (2.8%) and 32 (45.7%) resistance rates had the highest effect. Among all the isolates, 55 (78.5%), 7 (10%), and 3 (4.3%) were strong, moderate, and weak biofilm producers, respectively. The frequency rates of biofilm-related genes were 64 (91.4%), 70 (100%), 56 (80%), and 22 (31.42%) for bap, ompA, csuE, and blaPER1 , respectively. Overexpression of pmrA and pmrB genes was observed in two colistin-resistance isolates, but the expression of these genes did not change in colistin-sensitive isolates. Additionally, 37 (52.8%) and 8 (11.4%) isolates belonged to groups 1 (ICII) and 2 (IC I), respectively. MLST analysis revealed a total of nine different sequence types that six isolates belonged to clonal complex 92 (corresponding to ST801, ST118, ST138, ST 421, and ST735). Other isolates were belonging to ST133 and ST216, and two colistin-resistant (Ab4 and Ab41) isolates were belonging to ST387 and ST1812. CONCLUSION: The present study revealed the presence of MDR and XDR A. baumannii isolates harboring biofilm genes and emergence of colistin-resistant isolates in Southwestern Iran. These isolates had high diversity, which was affirmed by typing techniques. The control measures and regular surveillance are urgently needed to preclude the spread of these isolates.

Prevalence of Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae Causing Bloodstream Infections in Cancer Patients from Southwest of Iran.

INTRODUCTION: This study aimed to evaluate the frequency rate of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE) causing bloodstream infections (BSIs) in cancer patients referred to one of the major referral hospitals in Ahvaz city, southwest Iran. MATERIALS AND METHODS: In this study, 1700 blood cultures were collected from 610 cancer patients suspected to have BSI from October 2016 to August 2017 referred to the Shafa cancer hospital, Ahvaz, southwest of Iran. The blood culture bottles were incubated aerobically at 35-37ºC for 24 hours and then sub-cultured on routine microbiology culture media. The bacterial colonies were identified using standard tests. The antibiotic susceptibility testing was achieved by the disc-diffusion method. The phenotypic detection of ESBLs was carried out by the combination disc-diffusion test (CDDT). Finally, the polymerase chain reaction (PCR) was performed to investigate the presence of bla TEM, bla CTX, bla SHV, and bla PER genes. RESULTS: The prevalence of BSI in cancer patients was 16.4% (100/610). Gram-negative rods with rate of 74% (74/100) were the most prevalent bacteria. The frequency of Enterobacteriaceae family was 21% including Escherichia coli (n: 8), Klebsiella pneumoniae (n: 6), Enterobacter spp. (n: 5), Citrobacter freundii (n: 1), and Serratia marcescens (n: 1). All isolates were multidrug-resistant (resistance to three or more antibiotics). The results of CDDT showed that 42.8% (9/21) of Enterobacteriaceae isolates had a positive ESBL test of which 100% (9/9) indicated positive band for at least one of the ESBL genes by PCR method. The bla CTX-M and bla TEM genes were detected in 38% (8/21) and 23.8% (5/21) of isolates, respectively, while the bla SHV and bla PER were not detected in any isolates. CONCLUSION: Based on the results, surveillance, and antibiotic stewardship programs should be implemented for cancer patients to prevent the spread of more ESBL-PE that have limited therapeutically choices.

Investigation of SCCmec types I-IV in clinical isolates of methicillin-resistant coagulase-negative staphylococci in Ahvaz, Southwest Iran.

Today methicillin resistant coagulase-negative staphylococci (MR-CoNS) are important in terms of causing significant nosocomial infections. Besides, MR-CoNS are confirmed as the reservoir of SCCmec elements that carry mecA (methicillin-resistant) gene. Hence, the present study was designed to evaluate the susceptibility pattern, prevalence and diversity of SCCmec types I, II, III, and IV in MR-CoNS strains. In this cross-sectional study, 44 clinical isolates of MR-CoNS were identified using the cefoxitin disc method and further confirmation by polymerase chain reaction (PCR) amplification of the mecA gene. Antimicrobial susceptibility of isolates was investigated by disc diffusion. The identification of CoNS was done by amplification and sequencing of the tuf gene. Multiplex PCR method was done for the determination of SCCmec types. In the present study, the Staphylococcus epidermidis and Staphylococcus haemolyticus were the most predominant isolates with a prevalence of 45.4%. The highest resistance rates were observed against erythromycin (84.1%) and clindamycin (75%). Multiplex PCR revealed the SCCmec type I as the predominant type in the present study. Our study showed that there was no significant relationship between the presence of different types of SCCmec elements and resistance to antibiotics. The present study highlighted a frequent prevalence of MR-CoNS harboring SCCmec type genes in Ahvaz, southwest of Iran. Thus, the molecular typing and periodical monitoring of their drug resistance pattern should be considered in national stewardship programs to designing useful antibiotic prescription strategies.

Application of tuf gene sequence analysis for the identification of species of coagulase-negative staphylococci in clinical samples and evaluation of their antimicrobial resistance pattern.

INTRODUCTION: Coagulase-negative staphylococci (CoNS) are normal inhabitants of human skin and mucous membranes. However, CoNS represent one of the major nosocomial pathogens, especially in immunocompromised patients. The increasing incidence of CoNS and mainly methicillin-resistant strains underlines the need for an accurate identification of Staphylococcus isolates at the species level. Analysis of the tuf gene proved to be an accurate tool for the species identification of CoNS. The aims of this study were to identify the CoNS species by tuf gene-based polymerase chain reaction method and sequencing, and to determine the frequency of CoNS clinical isolates resistant to methicillin (MRCoNS) and other antibiotics. METHODS: A total of 200 staphylococci isolates were collected from various clinical samples. Phenotyping methods were used for initial identification followed by polymerase chain reaction amplification of tuf gene with subsequent sequencing. The phylogenetic relationships among species were analyzed using the neighbor-joining method based on the partial gene sequence of tuf. Microbroth dilution test was used for screening methicillin resistance, and disk diffusion susceptibility testing was performed for evaluation of antibiotic resistance among the isolates. RESULTS: In the present study, 125 isolates were identified as CoNS; among them, Staphylococcus epidermidis 54(43.2%) and Staphylococcus haemolyticus 50 (40.0%) were demonstrated as the most prevalent species. Resistance to methicillin was detected in 54.4% of the CoNS based on microbroth dilution method. In disk diffusion susceptibility testing, the greatest resistance of CoNS was demonstrated for cefoxitin (65.4%), cotrimethoxazole (54.4%), and clindamycin (49.6%), while daptomycin (87.2%) and linezolid (83.2%) showed the greatest effectiveness for CoNS isolates. CONCLUSION: Our results confirmed the predominance of S. epidermidis and S. haemolyticus among CoNS isolates. The high prevalence of MRCoNS strains is a serious concern and strongly suggests the need for control program measures in our hospitals in order to reduce MRCoNS infections, especially in immunocompromised patients.

Antibacterial Effects of Chitosan, Formocresol and CMCP as Pulpectomy Medicament on Enterococcus faecalis, Staphylococcus aureus and Streptococcus mutans.

INTRODUCTION: During pulpectomy of primary teeth, cytotoxic medicaments such as formocresol or camphor mono-chlorophenol (CMCP) are used as medicaments. For the first time it is theorized that chitosan can substitute these traditional materials used in pulpectomy of infectious primary teeth. METHODS AND MATERIALS: This preliminary in vitro study consisted of two separate phases (n=75), each of which assessed the antibacterial effects of chitosan versus formocresol and CMCP and positive/negative controls (n=15) on three bacteria types [Enterococcus faecalis, Staphylococcus aureus, Streptococcus mutans, (n=5 per subgroup)]. Phases 1 and 2 concerned respectively with 1- and 7-day effects of these materials. Bacteria were cultured and injected into sterilized canals and colonies were counted. Medicaments were applied and colonies were re-counted after 1 day of treatment (phase 1). Specimens were re-sterilized and re-randomized, and used for phase 2, in which the same procedures were performed for a 7-day period. Effects of agents on bacteria were analyzed statistically (Kruskal-Wallis α=0.05 and Mann-Whitney α=0.017). RESULTS: Treatments reduced bacterial count either after 1 or 7 days (P=0.000). Their effects on different bacteria types were not significant either after 1 or 7 days (P>0.48). Antibacterial efficacies of treatments (indicated by colony reduction) were significantly different, after 7 days (P=0.045). Antibacterial efficacy of chitosan was similar to that of formocresol or CMCP, in both phases [either after 1 or 7 days of treatment (P>0.017). Formocresol and CMCP had similar efficacies in either phase (P>0.017). CONCLUSIONS: This preliminary study confirmed the appropriate antibacterial efficacy of chitosan as a medicament in pulpectomy of infectious primary teeth.

The frequency of class1 and 2 integrons in Pseudomonas aeruginosa strains isolated from burn patients in a burn center of Ahvaz, Iran.

BACKGROUND: Pseudomonas aeruginosa is an opportunistic pathogen with the ability to cause severe nosocomial infections and remains a major problem in burn patients. This organism shows a remarkable antimicrobial resistance and is often resistant to multiple antibiotics. Integron genes as mobile genetic elements are playing an important role in the spread of P. aeruginosa antibiotic resistance. This study was aimed to investigate the occurrence of class 1, and 2 integron genes (int1, int2), among P. aeruginosa strains isolated from patients with burn infections. METHODS: In total 93 clinical isolates of P. aeruginosa were screened. The antimicrobial susceptibilities of 9 common antimicrobial agents were tested against the isolates using disk diffusion method. PCR amplification was performed on extracted DNAs for the detection of int1, and int2 genes using the set of specific primers. RESULTS: The majority of P. aeruginosa isolates were from wound infection (69.9%). In disk diffusion method, most isolates showed remarkable resistance to tested antibiotics with highest against gentamicin (94.62%) and ciprofloxacin (93.55%). PCR amplification revealed that 89(95.7%) of P. aeruginosa strains carried int1, but none of them harbored int2 genes. The distribution of int1 gene was highest in blood (100%), followed by wound isolates (95.38%). CONCLUSIONS: We demonstrated a high antimicrobial resistance among P. aeruginosa isolates in our setting. int1 was prevalent and seems to play an important role in multidrug resistance among the isolates. So, performance of antibiotic surveillance programs is necessary for choosing the appropriate therapy and management of infection control practices.

The frequency of genes encoding exotoxin A and exoenzyme S in Pseudomonas aeruginosa strains isolated from burn patients.

BACKGROUND: Pseudomonas aeruginosa infections have emerged as a major infectious disease threat in recent decades with infection particularly in immunocompromised hosts. P. aeruginosa possesses several virulence factors with involvement in pathogenesis. The aim of this study was to examine the prevalence of virulence genes of toxA and toxS and to analyze their relation to antimicrobial resistance of the isolates. METHODS: In total 185 clinical isolates of P. aeruginosa were collected from burn patients. Antimicrobial susceptibility testing was done by disk diffusion method. PCR amplification was performed on extracted DNA from the isolates and the presence of encoding genes for exotoxin A (toxA) and exoenzyme S (toxS) were investigated by using specific primers. RESULTS: In disk diffusion method, the isolates showed high sensitivity to colistin sulfate (100%) followed by imipenem (41.9%). The most prevalent resistance was seen against ceftazidime (90.5%) and gentamicin (88.5%). Multidrug resistance (MDR) demonstrated in 113 isolates (76.35%). According to PCR amplification, 133 (89.8%) and 127 (85.8%) isolates possessed toxA and toxS genes respectively. The frequencies of genes among MDR strains were 102 (76.6%) for toxA and 98 (77.1%) for toxS. Eighty five MDR isolates possessed both genes (73.9%). The non-MDR strains (23.65%), harbored lower prevalence of simultaneous toxA and toxS genes (26%) compared to MDR strains. CONCLUSION: The present study established a higher frequency of MDR among P. aeruginosa isolates from burn patients. It was found that the frequency of both toxA &S genes were significantly higher in MDR strains P. aeruginosa strains.

Methicillin Resistant Staphylococci: Prevalence and susceptibility patterns in a burn center in Ahvaz from 2013-2014.

BACKGROUND AND OBJECTIVES: Methicillin resistance Staphylococcus aureus (MRSA) and coagulase negative staphylococci (MRCoNS) have recognized as the major cause of nosocomial infections that threat the burn patient's life. The aims of this study were to determine the frequency of MRSA and MRCoNS and their antibiotic resistance patterns among burn patients in a burn center in Ahvaz, Iran. MATERIAL AND METHODS: A total of 340 clinical specimens: (80%) wound and (20%) blood were obtained from patients in Taleghani burn hospital during February 2013-2014. Staphylococci species identification and antibiogram were performed by standard procedures using disk diffusion method. The Methicillin resistance strains were detected by Etest and PCR using mecA specific primers. RESULTS: Out of 30.2% (103) isolates that were recognized as staphylococci, 82 % (84) and 18% (19) were identified as S. aureus and coagulase negative staphylococci (CoNS) respectively. Resistance to methicillin was detected in 60% and 63% of the S. aureus and CoNS isolates respectively. Seven different antimicrobial resistance patterns observed among methicillin resistant staphylococci. The MRSA and MRCoNS strains showed closed resistance phenotypes. All the methicillin resistant isolates showed a high rate resistance to the other studied antibiotics in comparison to methicilin sensitive isolates. Vancomycin and imipenem showed the greatest effect against methicillin resistant isolates. During 8 years in the studied burn hospital, no significant changes in the methicillin resistance staphylococci frequency were detected. CONCLUSION: The presence of multi resistant MRSA and MRCoNS strains is cause of concern in burn hospitals. Vancomycin remains as a drug of choice for methicillin resistance staphylococci infections.

Sub-Conjunctival Injection of Antibiotics vs. Povidone-Iodine Drop on Bacterial Colonies in Phacoemulsification Cataract Surgery.

BACKGROUND: Postoperative endophthalmitis is one the most serious complications of cataract surgery. The majority of causative organisms in this destructive infection come from the patient's own periocular flora. Efforts have been made to reduce the virulence of organisms in the eyelid and conjunctiva with perioperative topical antibiotics, preparation of surgical field, covering eyelids and conjunctival surface with 5% povidone-iodine solution and intracameral antibiotics at the time of surgery to minimize the risk of endophthalmitis. OBJECTIVES: We assessed the effect of subconjunctival injection of cefazolin and pouring povidone-iodine on the conjunctiva bacterial colony forming units (CFU) in phacoemulsification cataract surgery. PATIENTS AND METHODS: In this prospective, randomized, double-blind clinical trial, 122 patients having phacoemulsification cataract surgery with clear corneal incision and topical anesthesia were randomized into two groups including group 1 (subconjunctival injection of cefazolin) and group 2 (recipients of a drop of povidone-iodine). Cultures were collected from the bulbar conjunctiva at the injection site and from the corresponding location in the patient's eye, three different times. RESULTS: The mean of eyelid samples on blood and chocolate agars, on the day after compared to the day before the surgery in group 1 showed a 52% and 56% reduction. These values were 58% and 50% in group 2 (P < 0.05). The mean CFU of conjunctiva before and at the end of surgery on blood and chocolate agars showed 57% and 56% reduction in group one and 51% and 52% reduction in group 2 (P < 0.05). While comparing mean CFU of conjunctiva at the end and one day post-surgery (interval of 14 ± 2 hours) showed 27% and 27% increase in group 1 and 20% and 21% increase in group 2 (P < 0.05), which reflects conjunctival flora proliferation during the early postoperative period. CONCLUSIONS: Due to the good tolerance of patients towards topical anesthesia, pouring a drop of povidone-iodine 10% seems to be a simple and acceptable method to reduce the growth of microorganisms of the conjunctiva.

The prevalence of methicillin resistant Staphylococcus aureus (MRSA) isolates with high-level mupirocin resistance from patients and personnel in a burn center.

The rate of the MRSA strains, particularly at burn centers, is increasing worldwide. Detection of mupirocin resistance MRSA strains in the burn centers particularly from personnel will help to control these strains. For this purpose, a total of 116 Staphylococcus aureus isolates from the patients (burns) and personnel (nostrils) in Ahvaz Taleghani hospital (Iran) were investigated. The methicillin and mupirocin resistant isolates were detected by multiplex amplification of the mecA and ileS-2 genes. The mecA was found among 80% of isolates. The rates of mupirocin resistant strains among personnel and patients were 70% and 6%, respectively. The carriage rates of the S. aureus, MRSA and MRSA with high-level mupirocin resistance in the personnel were 40%, 34% and 28%, respectively. In conclusions, the high prevalence of MRSA strains in the patients showed the potential outbreak of the MRSA in the burn center and highlighted the need of antibiotic susceptibility monitoring of MRSA. Moreover being personnel as a main reservoir in terms of MRSA strains with high-level mupirocin resistance emphasizes the screening of the personnel in terms of the MRSA in the healthcare system especially in the burn center.