Biology-Oriented Drug Synthesis (BIODS) Approach towards Synthesis of Ciprofloxacin-Dithiocarbamate Hybrids and Their Antibacterial Potential both in Vitro and in Silico.

A novel series of ciprofloxacin-dithiocarbamate hybrids 7a - 7l were designed, synthesized, and evaluated against Gram-positive and Gram-negative bacteria. A significant part of the title compounds showed considerable antibacterial activity against Gram-positive species. The most potent compound against Gram-positive bacteria was 2-chloro derivative 7h and the most potent derivative against Gram-negative bacteria was 3-chloro compound 7i. In vitro antibacterial evaluation of compound 7h against clinically isolated bacteria methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) showed that this compound acted better than ciprofloxacin against the latter bacteria. Docking study of compound 7h in the active site of S. aureus DNA gyrase revealed that this ciprofloxacin-dithiocarbamate derivative interacted with the main components of the active site of the enzyme.

Integron types, gene cassettes and antimicrobial resistance profile of Acinetobacter baumannii isolated from BAL samples in Babol, north of Iran.

Multi-drug resistant isolates of Acinetobacter baumannii have created therapeutic problems worldwide. This current study was intended to determine the Integron types, gene cassettes and antimicrobial resistance profile of A. baumannii isolated from BAL samples in Babol, north of Iran. During a 15-month period, 35 A. baumannii isolates were studied. Different classes of antimicrobial agents were used to determine the resistance ratios. Multiplex-PCR was used to detect different types of integrons and associated gene cassettes. The resistance rates to GM, FEP, AK, TOB, CP, PIP, SAM, IPM, SXT, CTX, CAZ, CL, TIM, MEM, and TZP were 85.7%, 100%, 91.4%, 68.5%, 94.3%, 88.5%, 97.1%, 94.3%, 100%, 100%, 100%, 0.0%, 91.4%, 94.3% and 91.4%, respectively. The distribution analysis of int genes showed that 25.7%, 88.6% and 28.6% of isolates carried the intI, intII and intIII genes, respectively. The prevalence of aadB, dfrA1, bla-OXA30 and aadA1 genes were 94.3%, 77.1%, 40% and 5.7%, respectively. The current study showed that a high level of A. baumannii isolates harbor integrons in our therapeutic center, which may lead to distribution of multiple antimicrobial resistance. The different types of gene cassette arrays in the present study highlight the important role of geographical features in MDR isolates dissemination which could be credited to different profiles of drug consumption in different areas. The findings emphasized that the need for continuous surveillance to prevent distribution of multidrug resistance among A. baumannii strains in Iran.

Determination of Helicobacter pylori virulence-associated genes in duodenal ulcer and gastric biopsies.

Background:Helicobacter pylori (H. pylori or Hp) has been strongly associated with the peptic ulcer diseases, chronic gastritis, ulcers, and gastric cancer. Genes associated with pathogenicity have been designated for H. pylori, and some of them appear to be related to more severe clinical consequences of the infection. The present study was conducted to determine cagA, vacA, cagE, iceA1, oipA, and iceA2 genes in H. pylori strains isolated from gastroduodenal patients, who referred to Shariati hospital in Tehran, Iran. Methods: Gastric biopsy specimens were collected during endoscopy from patients, who referred to the Shariati hospital in Tehran, Iran during January and November 2015. After isolation of H. pylori from the biopsy culture, genomic DNA was extracted and subsequently used to identify H. pylori and virulence genes using specific primers. Results: The isolation rate of H. pylori strains was 65.7% (169/257). The frequency of cagA, vacA, cagE, iceA1, oipA, and iceA2 was 143 (% 84.6), 169 (100%), 131 (77.5%), 97 (57.3%), 89 (52.6%), and 72 (42.6%), respectively. Conclusion: In this study, a significant difference was observed between investigated genes and strains isolated from PUD and GC patients (p<0.05).

Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species.

OBJECTIVE: Rapid, reliable, and affordable detection of Brucella species via the molecular methods remains a challenge. In recent years, loop-mediated isothermal amplification (LAMP) is a functional nucleic acid amplification technique offering a substitute to polymerase chain reaction (PCR). So, we compared the LAMP assay with the conventional PCR for the identification of common Brucella species in Iran. In this study, LAMP assay was comprehensively evaluated against the common PCR method. A group of specific LAMP primers were used to amplify a highly specific fragment from the sequence of the Brucella abortus, bcsp31 gene. Sensitivity and specificity values of tests were done with a set of 78 (50 Brucella and 28 non-Brucella) strains. RESULTS: A dilution series of B. abortus DNA indicated that the LAMP reaction could reliably detect 10 (fg/µl) DNA target copies per reaction within 36 min, which is 10 times greater than the PCR assay. In summary, we conclude that LAMP assay provide accurate and fast test results to identify of common Brucella species in low-complexity labs, mainly in low and lower middle income countries.

High-Level Aminoglycoside Resistance in Enterococcus Faecalis and Enterococcus Faecium; as a Serious Threat in Hospitals.

AIMS AND OBJECTIVES: The present work aimed to evaluate the frequency of aminoglycoside- modifying enzymes encoding genes in the E. faecalis and E. faecium and their antibiotic resistance profile. METHODS: A total of 305 different clinical samples were subjected for identification and antibiotic susceptibility test. The high-level aminoglycoside resistance was identified by MIC and Kirby Bauer disc diffusion method. The prevalence of aac (6')-Ie-aph (2'')-Ia, aph (3')-IIIa and ant (4')- Ia genes was determined by multiplex- PCR. In total, 100 enterococci strains were isolated. The prevalence of E. faecalis and E. faecium isolates was 78% and 22%, respectively. RESULTS: All isolates were susceptible to linezolid. So, all E. faecalis were susceptible to vancomycin but, 36.4% of E. faecium were resistant to it. The prevalence of multiple drug resistance strains was 100% and 67.9% of E. faecium and E. faecalis, respectively. High-level-gentamicin and streptomycin resistant rates were as follows; 26.9% and 73.1% of E. faecalis and 77.3% and 90.1% of E. faecium. Conclucion: The results of the current study showed a high frequency of aac (6')-Ie-aph (2'')-Ia genes among enterococcal isolates. A high rate of resistance to antimicrobials in Enterococcus is obviously problematic, and a novel policy is needed to decrease resistance in these microorganisms.

Role of Clotrimazole in Prevention of Recurrent Otomycosis.

Otomycosis is one of the relatively common diseases in the world which is caused by different fungi especially saprophytes. Concerning the relapse of this disease in a number of individuals, the present study was performed to evaluate the inhibitory effect of clotrimazole drop in the relapse of otomycosis. Clinical samples were taken by an ENT specialist from patients suspicious of having otomycosis. A part of these samples were stained, and others were cultured. The diagnosis of otomycosis was made on the basis of the recognizable and characteristic appearance of fungal hyphae or mycelium and fruiting bodies and/or conidiophores under microscopic examination. Patients with suspected otomycosis are not at risk of recurrence after treatment with clotrimazole drops. Out of the 161 individuals in whom definite diagnosis of otomycosis was made, the most affected individuals were, in the age range of 40-49 years, women, urban citizens, and housewives. Pruritus and diminished hearing were the main complaints of the patients. Aspergillus niger and A. flavus as well as Candida albicans were the main causes of the disease. The relapse of disease was observed in only five patients (3.1%), where A. niger was the main fungus. Most relapses were observed in women and in those with diminished hearing, manipulating the ears, ulcers in the canal, and tympanum. Our results suggested that usage of clotrimazole can be effective in reducing the relapse of otomycosis, and concerning the high cost of treating otomycosis while the low cost of using clotrimazole, usage of this drop is recommended to reduce the relapse of otomycosis.

Analysis of Resistance to Macrolide-Lincosamide-Streptogramin B Among mecA-Positive Staphylococcus Aureus Isolates.

OBJECTIVES: Genetic determinants conferring resistance to macrolide, lincosamide, and streptogramin B (MLSB) via ribosomal modification such as, erm, msrA/B and ereA/B genes are distributed in bacteria. The main goals of this work were to evaluate the dissemination of MLSB resistance phenotypes and genotypes in methicillin-resistant Staphylococcus aureus (MRSA) isolates collected from clinical samples. METHODS: A total of 106 MRSA isolates were studied. Isolates were recovered from 3 hospitals in Tehran between May 2016 to July 2017. The prevalence of MLSB-resistant strains were determined by D-test, and then M-PCR was performed to identify genes encoding resistance to macrolides, lincosamides, and streptogramins in the tested isolates. RESULTS: The frequency of constitutive resistance MLSB, inducible resistance MLSB and MSB resistance were 56.2%, 22.9%, and 16.6%, respectively. Of 11 isolates with the inducible resistance MLSB phenotype, ermC, ermB, ermA and ereA were positive in 81.8%, 63.6%, 54.5% and 18.2% of these isolates, respectively. In isolates with the constitutive resistance MLSB phenotype, the prevalence of ermA, ermB, ermC, msrA, msrB, ereA and ereB were 25.9%, 18.5%, 44.4%, 0.0%, 0.0%, 11.1% and 0.0%, respectively. CONCLUSION: Clindamycin is commonly administered in severe MRSA infections depending upon the antimicrobial susceptibility findings. This study showed that the D-test should be used as an obligatory method in routine disk diffusion assay to detect inducible clindamycin resistance in MRSA so that effective antibiotic treatment can be provided.

Outer Ear Infections in Iran: A Review.

BACKGROUND: Otitis externa is the fungal and bacterial infection of the outer ear. AIM: We aimed to investigate the published papers about the outer ear infections in Iran and suggest standardised investigations and treatments. METHODS: We used different electronic databases like PubMed, Scopus, Web of Science, Iranmedex, Google Scholar, and Magiran with specific keywords. RESULTS: We obtained forty published full-text articles for review of data. Our results indicated the women were more infected than men. The ages of patients were < 1-81 years. As clinically symptoms, itching and Feel the ear fairy were the most common presenting complaints in most cases. Most infections were the pure bacterial and fungal origin, respectively. However, some of the studies were mixed fungal-bacterial infections - Pseudomonas spp. And Aspergillus niger were the most common bacteria and fungi isolates respectively in Iranian patents. CONCLUSION: Fungal and bacterial specific cultures may be recommended, and anti-fungal drugs may be added, to treatment regimens in patients with otitis externa to reduce the clinical symptoms.

Evaluation of antibacterial properties of hydroxyapatite/bioactive glass and fluorapatite/bioactive glass nanocomposite foams as a cellular scaffold of bone tissue.

AIMS AND OBJECTIVES: Infection is a serious problem for patients after implantation surgery, which is difficult to treat with antibiotic therapy. The present study was developed to evaluate and compare the antibacterial properties of hydroxyapatite/bioactive glass (HA/BG) and fluorapatite/bioactive glass (FA/BG) nanocomposite foams as a cellular scaffold for use in bone defects by two macrodilution and disk diffusion methods. MATERIALS AND METHODS: Staphylococcus aureus, Enterococcus faecalis, and Streptococcus mutans were cultured in brain heart infusion broth medium with nanocomposite powder for 5 days, and their bioactivity levels were evaluated by daily culturing on solid agar medium plates. To carry out the disk diffusion test, a disc form of nanocomposite foams was used on agar medium with 48 h incubation. RESULTS: None of two nanocomposites even at their highest concentration (200 mg/mL) did not prevent the growth of two Staphylococcus aureus and Enterococcus faecalis microorganisms. However, HA/BG nanocomposite on the 3rd day at a concentration of 200 mg/mL and on 4th and 5th day at a concentration of 100 mg/mL and FA/BG nanocomposite on the 4th day at a concentration of 100 mg/mL and on the 5th day at a concentration of 50 mg/mL could be able to kill Streptococcus mutans microorganism. In the disc diffusion test, none of the nanocomposites could create a nongrowth zone. Both tested biomaterials showed increased antibacterial properties over time and concentration increase. CONCLUSION: HA/BG and FA/BG nanocomposites, due to their biocompatibility and antimicrobial properties, are good choices for implantation instead of damaged bone tissue in tissue engineering.

Integron types, antimicrobial resistance genes, virulence gene profile, alginate production and biofilm formation in Iranian cystic fibrosis Pseudomonas aeruginosa isolates.

Cystic fibrosis (CF) patients commonly suffer from continuous and recurrent lung infections caused by Pseudomonas aeruginosa, the dominant pathogen in CF airways. This study aimed to determine the integron types, gene cassettes, virulence determinants, ß-lactam resistance genes, biofilm formation and alginate production in P. aeruginosa isolated from Iranian CF patients. A total of 143 P. aeruginosa isolates were obtained from CF patients. Susceptibility of isolates to different antimicrobials was evaluated by disc diffusion method. ESBL, MBL and KPC production was assessed. Congo red agar and tissue culture plates were used for evaluation of biofilm formation. Alginate production was determined using the Carbazole assay. Integrase genes, resistance determinants (ESBLs, MBLs and KPC) and genes encoding virulence factors were evaluated by PCR. All isolates were susceptible to colistin, piperacillin-tazobactam and ticarcillin; 8.4% of isolates were considered as MDR phenotype. Out of 6.3% IPM-resistant isolates, prevalence of virulence genes was as follows: lasB (100%) and plcB (100%), plcH (96.5%). Biofilm formation and alginate production ability were found in 54.5% of isolates. The prevalence of the alginate-encoding genes was 92.3%, 86.7% and 67.1% for algD, algU and algL genes, respectively. PpyR, pslA and pelA genes were detected in 98.6%, 89.5% and 57.3% of the isolates, respectively. The high prevalence of colonization in CF lungs may increase the pathogenicity of P. aeruginosa due to their adhesion and protective properties caused by biofilm- and alginate-production. LasB, plcB, plcH, exoS, toxA, algD, ppyR and pslA genes were predominant in CF P. aeruginosa strains.

Antimicrobial effect, frictional resistance, and surface roughness of stainless steel orthodontic brackets coated with nanofilms of silver and titanium oxide: a preliminary study.

Nano-silver and nano-titanium oxide films can be coated over brackets in order to reduce bacterial aggregation and friction. However, their antimicrobial efficacy, surface roughness, and frictional resistance are not assessed before. Fifty-five stainless-steel brackets were divided into 5 groups of 11 brackets each: uncoated brackets, brackets coated with 60 µm silver, 100 µm silver, 60 µm titanium, and 100 µm titanium. Coating was performed using physical vapor deposition method. For friction test, three brackets from each group were randomly selected and tested. For scanning electron microscopy and atomic-force microscopy assessments, one and one brackets were selected from each group. For antibacterial assessment, six brackets were selected from each group. Of them, three were immediately subjected to direct contact with S. mutans. Colonies were counted 3, 6, 24, and 48 h of contact. The other three were stored in water for 3 months. Then were subjected to a similar direct contact test. Results pertaining to both subgroups were combined. Groups were compared statistically. Mean (SD) friction values of the groups 'control, silver-60, silver-100, titanium-60, and titanium-100' were 0.55 ± 0.14, 0.77 ± 0.08, 0.82 ± 0.11, 1.52 ± 0.24, and 1.57 ± 0.41 N, respectively (p = .0004, Kruskal-Wallis). Titanium frictions were significantly greater than control (p < .05), but silver groups were not (p > .05, Dunn). In the uncoated group, colony count increased exponentially within 48 h. The coated groups showed significant reductions in colony count (p < .05, two-way-repeated-measures ANOVA). In conclusions, all four explained coatings reduce surface roughness and bacterial growth. Nano-titanium films are not suitable for friction reduction. Nano-silver results were not conclusive and need future larger studies.

Integron-Mediated Antibiotic Resistance in Acinetobacter baumannii Isolated from Intensive Care Unit Patients, Babol, North of Iran.

BACKGROUND: We investigated the integron types and their relation with antibiotic resistance among A. baumannii isolates collected from intensive care unit patients, Babol, north of Iran. METHODS: In this cross-sectional study, a total of 73 bronchoalveolar lavage samples were obtained from patients in ICU. Susceptibility testing was performed by disk diffusion method. Types of integrons were identified by an integrase gene PCR. RESULTS: In total, 47.9% A. baumannii isolates were recovered from the BAL samples. All isolates were resistant to ceftazidime. 91.4% and 58.3% of isolates were MDR and XDR, respectively. The rate of colistin resistance with the E-test was 5.7%. Molecular analysis of class I, II, and III integrons showed that 25.7%, 88.6%, and 28.6% of the isolates carried the intI, intII, and intIII genes, respectively. DISCUSSION: Our results show that different classes of integrons are commonly spread among A. baumannii strains and these genomic segments can play an important role in the acquisition of MDR and XDR phenotypes. So monitoring drug resistance in A. baumannii isolates with the use of int gene PCR is very important to plan specific infection control measures to prevent the spread of MDR-AB and XDR-AB in Iran's hospitals.

Evaluation of the eukaryotic expression of mtb32C-hbha fusion gene of Mycobacterium tuberculosis in Hepatocarcinoma cell line.

BACKGROUND AND OBJECTIVES: HBHA and Mtb32C have been isolated from culture supernatants of Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium bovis (M. bovis) and their immunogenicity previously studies have been confirmed. In this study, capability of constructed vector containing two mycobacterial immunodaminant antigens (Mtb32C-HBHA), in producing new chimeric protein under the in-vitro condition was examined. MATERIALS AND METHODS: In present study Huh7.5 cells was transfected with Mtb32C-HBHA -pCDNA3.1+ recombinant vector using the calcium phosphate method and expression of chimeric protein was assessed by RT-PCR and Western blot methods. RESULTS: Results of RT-PCR and Western blot showed expression of 35.5 KD recombinant protein (Mtb32C-HBHA) in this cell line. CONCLUSION: The constructed vector can produce two highly immunogenic antigens that fusion of them to gather makes chimeric antigen with new traits. Other attempts are needed to evaluate specific properties of this new antigen such as molecular conformation modeling and immunologic characteristics in future studies.

Anti-Streptococcus mutans property of a chitosan: Containing resin sealant.

OBJECTIVE: This study sought to assess the inhibitory effect of chitosan-containing sealants against Streptococcus mutans. MATERIALS AND METHODS: The antibacterial activity of the resin sealant was evaluated by direct contact test following the addition of 0, 1, 2, 3, 4, and 5 wt% chitosan. At 3, 6, 9, 24 and 48 h, 1 and 3 months, 10 µl of the microbial suspension in contact with resin sealant was cultured to count the number of colonies. Data were analyzed by one-way one-way analysis of variance (ANOVA), repeated measures ANOVA, and Scheffe test. RESULTS: The minimum inhibitory concentration of chitosan against S. mutans was 2 wt%. At 3 h, bacterial count in the presence of 2-5 wt% chitosan was significantly lower than that at 0 and 1 wt% (P < 0.05). However, this difference in bacterial count between 2 and 3 wt% chitosan and between 4 and 5 wt% chitosan was not significant. At 6 h, the difference in bacterial count between 3 and 4 wt% chitosan was not significant, whereas the remaining groups were significantly different in terms of bacterial count at this time (P < 0.05). At the remaining time points, significant differences were found between 2 wt% chitosan and higher concentrations (P < 0.05). CONCLUSION: Sealants containing 2-5 wt% chitosan show an antimicrobial property that is intensified by increasing the concentration of chitosan.

Detection of Merkel Cell Polyomavirus and Human Papillomavirus in Esophageal Squamous Cell Carcinomas and Non-Cancerous Esophageal Samples in Northern Iran.

Human papillomavirus (HPV) infection is one of the hypothesized causes of esophageal squamous cell carcinoma (ESCC), but the etiological association remains uncertain. It was postulated that other infectious agents together with HPV may increase the risk of ESCC. The current investigation aimed to explore the presence of a new human tumor virus, Merkel cell polyomavirus (MCPyV), together with HPV in ESCC tumors and non-cancerous esophageal samples in northern Iran. In total, 96 esophageal samples (51 with ESCC, and 45 without esophageal malignancy) were examined. HPV DNA was detected in esophageal specimens of 16 out of the 51 ESCC cases (31.4 %) and 20 out of the 45 non-cancerous samples (44.4 %). Untypable HPV genotypes were recognized in high rates in cancerous (75.0 %) and non-cancerous (55.0 %) esophageal specimens. MCPyV DNA was detected in esophageal specimens of 23 out of the 51 ESCC cases (45.1 %) and 16 out of the 45 non-cancerous samples (35.6 %). The mean MCPyV DNA copy number was 1.0 × 10(-5) ± 2.4 × 10(-5) and 6.0 × 10(-6) ± 1.3 × 10(-5) per cell in ESCC cases and non-cancerous samples, respectively. There was no statistically significant difference between cancerous and non-cancerous samples regarding mean MCPyV DNA load (P = 0.353). A bayesian logistic regression model adjusted to the location of esophageal specimen and MCPyV infection, revealed a significant association between HPV and odds of ESCC (OR, 2.45; 95 % CI: 1.01-6.16). This study provides the evidence of the detection of the MCPyV DNA at a low viral copy number in cancerous and non- cancerous esophageal samples.

Bacterial Otitis Externa in Patients Attending an ENT Clinic in Babol, North of Iran.

BACKGROUND: Acute otitis externa, an inflammatory condition of the external auditory canal, is a common clinical problem in general medicine. OBJECTIVES: This study aimed to determine the etiology of otitis externa in patients from the Mazandaran province, north of Iran, which has a humid climate, as humidity can affect the prevalence of pathogenic microorganisms. PATIENTS AND METHODS: This cross-sectional study involved 116 patients with otitis externa. Two sets of samples were collected from their ears; one set was used for slide preparations, and the other for microbial culturing. After culturing, the microorganisms were identified by conventional methods. RESULTS: Patients between 35 and 44 years of age were most frequently affected (25.00%) by otitis externa (average age, 43.87 ± 18.08 years). Moreover, women (54.31%) were more frequently affected than men (45.69%). Upon direct investigation, Gram-positive bacilli were the most commonly identified microorganisms (22.41%). Furthermore, Bacillus spp. and coagulase-negative staphylococci (22.41% and 19.83%, respectively), were the organisms most frequently identified from cultures of otitis externa samples. CONCLUSIONS: Direct examination and culture showed that a mixed infection of fungi and bacteria is the most common cause of otitis externa. The present study revealed that Bacilli spp. were the most abundant bacteria isolated from patients with otitis externa. Thus, it is recommended that both organisms should be considered as etiologic agents in protocols for treatment of otitis externa.

Evaluation of Antibacterial Effects of Silver-Coated Stainless Steel Orthodontic Brackets.

OBJECTIVES: White spots and enamel demineralization around orthodontic brackets are among the most important complications resulting from orthodontic treatments. Since the antibacterial properties of metals and metallic particles have been well documented, the aim of this study was to assess the antibacterial effect of stainless steel orthodontic brackets coated with silver (Ag) particles. MATERIALS AND METHODS: In this study, 40 standard metal brackets were divided into two groups of 20 cases and 20 controls. The brackets in the case group were coated with Ag particles using an electroplating method. Atomic force microscopy and scanning electron microscopy were used to assess the adequacy of the coating process. In addition, antibacterial tests, i.e., disk diffusion and direct contact tests were performed at three, six, 24, and 48 hours, and 15 and 30 days using a Streptococcus mutans strain. The results were analyzed using Student's t-test and repeated measures ANOVA. RESULTS: Analyses via SEM and AFM confirmed that excellent coatings were obtained by using an electroplating method. The groups exhibited similar behavior when subjected to the disk diffusion test in the agar medium. However, the bacterial counts of the Ag-coated brackets were, in general, significantly lower (P<0.001) than those of their non-coated counterparts. CONCLUSIONS: Brackets coated with Ag, via an electroplating method, exhibited antibacterial properties when placed in direct contact with Streptococcus mutans. This antibacterial effect persisted for 30 days after contact with the bacteria.

Wolbachia Endobacteria in Natural Populations of Culex pipiens of Iran and Its Phylogenetic Congruence.

BACKGROUND: Wolbachia are common intracellular bacteria that infect different groups of arthropods including mosquitoes. These bacteria modify host biology and may induce feminization, parthenogenesis, male killing and cytoplasmic incompatibility (CI). Recently Wolbachia is being nominated as a bio-agent and paratransgenic candidate to control mosquito borne diseases. METHODS: Here we report the results of a survey for presence, frequency, and phylogenetic congruence of these endosymbiont bacteria in Culex pipiens populations in Northern, Central, and Southern parts of Iran using nested-PCR amplification of wsp gene. RESULTS: Wolbachia DNA were found in 227 (87.3%) out of 260 wild-caught mosquitoes. The rate of infection in adult females ranged from 61.5% to 100%, while in males were from 80% to 100%. The Blast search and phylogenetic analysis of the wsp gene sequence revealed that the Wolbachia strain from Iranian Cx. pipiens was identical to the Wolbachia strains of supergroup B previously reported in members of the Cx. pipiens complex. They had also identical sequence homology with the Wolbachia strains from a group of distinct arthropods including lepidopteran, wasps, flies, damselfly, thrips, and mites from remote geographical areas of the world. CONCLUSION: It is suggested that Wolbachia strains horizontally transfer between unrelated host organisms over evolutionary time. Also results of this study indicates that Wolbachia infections were highly prevalent infecting all Cx. pipiens populations throughout the country, however further study needs to define Wolbachia inter-population reproductive incompatibility pattern and its usefulness as a bio-agent control measure.

Prevalence, and virulence determination of Listeria monocytogenes strains isolated from clinical and non-clinical samples by multiplex polymerase chain reaction.

INTRODUCTION:: This study aimed to determine the prevalence, and virulence factors of Listeria monocytogenes isolated from various samples by multiplex polymerase chain reaction (MPCR). METHODS:: A total of 617 isolates were obtained and MPCR was employed for detection of the inlA, inlC, and inlJ genes. RESULTS:: L. monocytogenes was detected in 46 (7.45%) of the 617 specimens. inlA, inlC, and inlJ were detected in 100%, 76.26%, and 71% isolates, respectively. CONCLUSIONS:: This study validated MPCR in the analysis and rapid detection of L. monocytogenes. The role of the genes in pathogenesis of the strains can also be affirmed.

Nosocomial emerging of (VIM1) carbapenemase-producing isolates of Klebsiella pneumoniae in North of Iran.

BACKGROUND AND OBJECTIVES: The rapid emergence and dissemination of carbapenemase-producing Klebsiella pneumoniae strains and other members of the Enterobacteriaceae poses a considerable threat to the care of hospitalized patients and to public health. The aim of this study was to determine the frequency of metallo-ß-lactamases (MBL) and VIM-1 gene in multidrug-resistant strains of K. pneumoniae. METHODS: 50 isolates of non - duplicated K. pneumoniae cultured from patients at intensive care units were tested for their susceptibilities to 13 different antibiotics using microbroth dilution assay. Isolates showing resistance to at least one of the carbapenems were checked for production of metallo-ß-lactamase (MBLs) using imipenem-EDTA synergy tests. PCR was used to detect the gene encoding VIM-1 metallo-ß-lactamase (MBL). RESULTS: Of 50 clinical isolates, 26 (52%) were resistant to imipenem in disk diffusion method. Using imipenem-EDTA synergy tests, production of MBL was detected in 15 (30%) isolates. PCR assay showed that 15 isolates were positive for VIM and these included 10 and 5 isolates showing positive and negative results in phenotypic method of MBL detection test respectively. Amikacin was found as the most effective antibiotic against the MBL producers in this study. CONCLUSION: The emergence of bla(VIM-1) producing K. pneumoniae in North of Iran is concerning. Microorganisms producing bla(VIM-1) constitute the prevalent multidrug-resistant population of K. pneumoniae in that region.