The Spread of Insertion Sequences Element and Transposons in Carbapenem Resistant Acinetobacter baumannii in a Hospital Setting in Southwestern Iran

Background@#Acinetobacter baumannii is one of the most important hospital pathogenic bacteria that cause infectious diseases. The present study aimed to determine the frequency of carbapenem resistance genes in association with transposable elements and molecular typing of carbapenem-resistant A. baumannii bacteria collected from patients in Shiraz, Iran. @*Materials and Methods@#A total of 170 carbapenem-resistant A. baumannii isolates were obtained from different clinical specimens in two hospitals. The minimum inhibitory concentrations (MIC) of imipenem were determined and the prevalence of OXA Carbapenemases, Metalloβ-lactamases genes, insertion sequences (IS) elements, and transposons were evaluated by the polymerase chain reaction (PCR) method. Finally, molecular typing of the isolates was performed by the Enterobacterial Repetitive Intergenic Consensus-PCR method. @*Results@#The MICs ranged from 16 to 1,024 µg/mL for imipenem-resistant A. baumannii isolates. Out of the 170 carbapenem resistant A. baumannii isolates, bla OXA-24-like (94, 55.3%) followed by bla OXA-23-like (71, 41.7%) were predominant. In addition, A. baumannii isolates carried bla VIM (71, 41.7%), bla GES (32, 18.8%), bla SPM (4, 2.3%), and bla KPC (1, 0.6%). Moreover, ISAba1 (94.2%) and Tn2009 (39.2%) were the most frequent transposable elements. Furthermore, (71, 44.0%) and (161, 94.7%) of the ISAba1 of the isolates were associated with bla OXA-23 and bla OXA-51 genes, respectively. Besides (3, 1.7%), (1, 0.6%) and (5, 2.9%) of bla OXA-23 were associated with IS18, ISAba4, and ISAba2, respectively. Considering an 80.0% cut off, clusters and four singletons were detected. @*Conclusion@#According to the results, transposable elements played an important role in the development of resistance genes and resistance to carbapenems. The results also indicated carbapenem-resistant A. baumannii bacteria as a public health concern.

Molecular characterization of a pigeon paramyxovirus type 1 virus isolated from Eurasian collared doves in Iran, 2017

In September 2017, an outbreak with high mortality, which showed the typical signs of ND, occurred among a flock of more than 2000 Eurasian collared doves in Konarak, southeast of Iran. A confirmed pigeon paramyxovirus type 1 strain was isolated from the brain tissues of the dead doves. The isolate, which was called Pigeon/Iran/Konarak/Barin/2017, was classified as a highly velogenic NDV. Complete genome sequencing and phylogenetic analysis showed that the isolate belonged to subgenotype XXI.2, which has never been reported from Iran before. The isolate had the highest homology (96.15%) with early 2010s Italian isolates.Further studies will be required to understand the diversity better.

Reduced Susceptibility to Biocides among Enterococci from Clinical and Non-Clinical Sources

Background@#Wide use of biocidal agents such as benzalkonium chloride (BCC) and chlorhexidine digluconate (CHX) in hospitals and non-hospital environments, has raised concerns over the emergence of non-susceptible strains. Efflux pumps are of known main mechanisms in biocide tolerance which have been rarely addressed in enterococci - members of gut microbiota which can cause serious problems particularly in hospitalized patients. The purpose of this study was to investigate the susceptibility of enterococci from different sources (clinical and fecal isolates) toward BCC and CHX, and its correlation with efflux associated genes. Also, possible link between biocide tolerance and antibiotic resistance was examined. @*Materials and Methods@#One hundred and four enterococcus isolates including clinical (n = 54) and fecal isolates (n = 50) were studied for susceptibility toward BCC, CHX, ciprofloxacin, gentamicin and vancomycin. Twelve efflux associated genes were investigated by polymerase chain reaction assay. @*Results@#In clinical isolates, reduced susceptibility to CHX and resistance to gentamicin and ciprofloxacin were significantly higher than fecal isolates. Vancomycin resistance was associated with increasing minimum inhibitory concentration of CHX. Among all investigated genes, only three ones, efrA, efrB and emeA were detected which were significantly associated with reduced susceptibility to CHX and were more frequent among clinical isolates. Also, high level resistance to gentamicin was significantly associated with the presence of efrA/B as well as with reduced susceptibility to CHX. @*Conclusion@#As expected, reduced susceptibility to CHX, was significantly higher in clinical isolates. However, the presence of a vancomycin-resistant enterococci among fecal isolates of healthy people which showed resistance/tolerance to studied antimicrobial agents, was unexpected and highlights the need to investigate other non-hospital environments to avoid dissemination of antimicrobial resistance. Correlation between reduced susceptibility to CHX and high level resistance to gentamicin, substantiates monitoring of biocide tolerance particularly in the healthcare settings to control the establishment of antimicrobial resistant strains.

Genotypic Diversity of Multidrug Resistant Shigella species from Iran

BACKGROUND: In many developing countries, shigellosis is endemic and also occurs in epidemics and treatment of multidrug-resistant (MDR) isolates are important. The aims of this study were to determine the antimicrobial susceptibility, prevalence of class 1 and 2 integrons and the clonal relatedness of isolates. MATERIALS AND METHODS: Antimicrobial susceptibility tests were performed by disc diffusion method. Polymerase chain reaction (PCR)-sequencing technique was employed for detection and characterization of integrons. The genetic relatedness was evaluated by using enterobacterial repetitive intergenic consensus (ERIC) PCR. RESULTS: There was a high percentage of resistance to trimethoprim-sulfamethoxazole (TMP/SMX) (93.7%), ampicillin (AMP) (87.3%), streptomycin (STR) (84.5%) and tetracycline (TET) (78.9%). Multidrug resistant phenotype was seen in 95.1% of total isolates. Most common MDR profile was TMP/SMX/STR/AMP resistant pattern. Among the 142 Shigella spp. analyzed in this study, 28 isolates were positive for class 1 integron with two types of gene cassette arrays (dfrA17/aadA5 = 31.7% and dfrA7 = 3.8%). The class 2 integron was more frequently detected among the isolates (94.7%) with dfrA1/sat1/aadA1 (69.4%) and dfrA1/sat1 (30.6%) gene cassettes. ERIC-PCR results showed 6, 5, 4 and 3 main genotypes among S. flexneri, S. sonnei, S. boydii and S. dysenteriae isolates, respectively. CONCLUSIONS: Our findings revealed that multidrug resistant Shigella species with high prevalence of class 2 integron were very common in Iran. In addition, ERIC-PCR patterns showed limited variety of clones are responsible for shigellosis in the region of the study.

Genotypic Diversity of Multidrug Resistant Shigella species from Iran

BACKGROUND: In many developing countries, shigellosis is endemic and also occurs in epidemics and treatment of multidrug-resistant (MDR) isolates are important. The aims of this study were to determine the antimicrobial susceptibility, prevalence of class 1 and 2 integrons and the clonal relatedness of isolates. MATERIALS AND METHODS: Antimicrobial susceptibility tests were performed by disc diffusion method. Polymerase chain reaction (PCR)-sequencing technique was employed for detection and characterization of integrons. The genetic relatedness was evaluated by using enterobacterial repetitive intergenic consensus (ERIC) PCR. RESULTS: There was a high percentage of resistance to trimethoprim-sulfamethoxazole (TMP/SMX) (93.7%), ampicillin (AMP) (87.3%), streptomycin (STR) (84.5%) and tetracycline (TET) (78.9%). Multidrug resistant phenotype was seen in 95.1% of total isolates. Most common MDR profile was TMP/SMX/STR/AMP resistant pattern. Among the 142 Shigella spp. analyzed in this study, 28 isolates were positive for class 1 integron with two types of gene cassette arrays (dfrA17/aadA5 = 31.7% and dfrA7 = 3.8%). The class 2 integron was more frequently detected among the isolates (94.7%) with dfrA1/sat1/aadA1 (69.4%) and dfrA1/sat1 (30.6%) gene cassettes. ERIC-PCR results showed 6, 5, 4 and 3 main genotypes among S. flexneri, S. sonnei, S. boydii and S. dysenteriae isolates, respectively. CONCLUSIONS: Our findings revealed that multidrug resistant Shigella species with high prevalence of class 2 integron were very common in Iran. In addition, ERIC-PCR patterns showed limited variety of clones are responsible for shigellosis in the region of the study.

Serogroups, subtypes and virulence factors of shiga toxin-producing Escherichia coli isolated from human, calves and goats in Kerman, Iran

Aim: The present study was conducted to detect the occurrence, serogroups, virulence genes and phylogenetic relationship of shiga toxin-producing Escherichia coli [STEC] in human, clave and goat in Kerman [southeast of Iran]

Background: STEC have emerged as the important foodborne zoonotic pathogens causing human gastrointestinal disease and confirming the risk to public health

Methods: A total of 671 fecal samples were collected from diarrheic patients [n=395] and healthy calves [n=156] and goats [n=120] and screened for the presence of stx gene. Furthermore, the prevalence of stx1 and stx2 variants, serotypes [O157, O145, O103, O26, O111, O91, O128, and O45], phylogenetic groups and the presence of ehxA, eae, hylA, iha and saa virulence genes were studied

Results: Prevalence of STEC in human diarrheic isolates was 1.3% [5 isolates], in claves was 26.3% [41 isolates] and in goats was 27.5% [33 isolates]. stx1 gene was the most prevalent variant and detected in 75 isolates. Furthermore, stx1c was the most predominant stx subtype, found in 56 isolates. The ehxA identified in 36 [45.6%] isolates, followed by iha 5 [6.3%], eaeA 4 [5.1%], hlyA 2 [2.5%] and saa 2 [2.5%]. Most of the isolates belonged to phylogroup B1. Only two O26 and one O91 isolates were detected in our study

Conclusion: Our results show that STEC strains were widespread among healthy domestic animals in the southeast of Iran

Two-dimensional and volumetric airway changes after bimaxillary surgery for class III malocclusion

OBJECTIVES: Any change in maxilla and mandible position can alter the upper airway, and any decrease in the upper airway can cause sleep disorders. Thus, it is necessary to assess airway changes after repositioning of the maxilla and mandible during orthognathic surgery. The purpose of this study was to evaluate linear and volumetric changes in the upper airway after bimaxillary surgery to correct class III malocclusion via cone-beam computed tomography (CBCT) and to identify correlations between linear and volumetric changes. MATERIALS AND METHODS: This was a prospective cohort study. CBCTs from 10 class III patients were evaluated before surgery and three months after. The Wilcoxon one-sample test was used to evaluate the differences in measurements before and after surgery. Spearman's rank correlation coefficient was used to test the correlation between linear and volumetric changes. RESULTS: The results show that the nasopharyngeal space increased significantly, and that this increase correlated with degree of maxillary advancement. No significant changes were found in volumes before and after surgery. A correlation was found between linear and volumetric oropharyngeal changes. CONCLUSION: Bimaxillary surgical correction of class III malocclusion did not cause statistically significant changes in the posterior airway space.

Molecular diagnosis and anti-microbial resistance patterns among Shigella spp. isolated from patients with diarrhea

Aim: This study aims to determine the serogroup distribution and molecular diagnosis, as well as antimicrobial resistance profiles among Shigella spp. isolated from patients with diarrhea in Kerman, southeast of Iran

Background: Shigella species are frequent cause of bacterial dysentery worldwide. Previous studies have been reported that S. sonnei and S. flexneri are the most prevalent serogroups in various parts of Iran

Patients and Methods: A total of 624 stool samples were randomly collected from patients with diarrhea from June 2013 to August 2014. Biochemical and serological characterizations were performed for identifying Shigella spp. In addition, the multiplex PCR assay was carried out for the detection and differentiation of three pathogenic Shigella spp. Antibiotic susceptibility testing was performed according to the Clinical Laboratory Standards Institute [CLSI] guidelines

Results: Fifty six [9%] Shigella strains were isolated from stool samples. The most common species were S. flexneri 31[55.4%], followed by S. sonnei 18[32.1%] and S. boydii 7[12.5%]. S. dysentery was not detected in the present study. All the isolates that identified by serological test as Shigella spp. were confirmed by the multiplex PCR method. The highest rate of resistance was observed for ampicillin and trimethoprim-sulphamethoxazole antibiotics with 52[92.9%] resistant, followed by tetracycline 44[78.6%] and cefotaxime 33[58.9%]. All Shigella isolates were susceptible to ciprofloxacin. A significant relationship was found between the Shigella species and cefotaxime resistance [p<0.05]

Conclusion: S. flexneri was found as the most prevalent serogroup causing shigellosis. The high rate of resistance to third-generation cephalosporins limits the treatment options available for the management of shigellosis in Kerman, Iran

[Effect of knee isokinetic extension training with maximum lateral tibial rotation on vastus amplitudes in patellofemoral pain syndrome patients]

Objective: Patellofemoral pain syndrome [PFPS] is one of the most common knee chronic disorders especially among females that is closely related to forces imbalance of vastus medial is oblique [VMO] and vastus lateral is [VL] muscles. The purpose of study was to examine the effect of knee isokinetic extension in maximum lateral tibia rotation on VMO and VL amplitudes in PFPS patients

Materials and Methods: Thirty-six women with PFPS participated voluntarily in this study and were randomly placed in one of three groups included the VMO selective isokinetic strengthening exercise, quadriceps general strengthening and control groups. Each exercise was performed for 8 weeks. Muscle RMS of VMO and VL and VMO/VL RMS ratio were recorded and calculated before and after training using of an 8-channels electromyography system. Data analysis was made by analyses of variance with repeated measures

Results: In baseline, VMO amplitude was less than VL in all groups [P0.05]. Yet, after interventions, VMO amplitude and VMO/VL amplitude ratio were more in selective group than in general and control groups [P

Conclusion: According to the study results, isokinetic extension training with maximum lateral tibia rotation and in close to knee full extension can be recommended as an appropriate training for improving VMO/VL electrical activity ratio and decreasing imbalance between lateral and medial vastus muscles amplitudes in patients with PFPS

Does sildenafil enhance the effect of tamsulosin in relieving acute urinary retention?

Objective To compare the safety and efficacy of combined therapy using sildenafil and tamsulosin for management of acute urinary retention (AUR) with tamsulosin alone in patients with benign prostate hyperplasia (BPH). Materials and Methods 101 patients were enrolled in a randomized placebo-controlled study from June 2009 to April 2012. Patients presenting with an initial episode of spontaneous AUR underwent urethral catheterization and then prospectively randomized to receive tamsulosin 0.4mg plus sildenafil 50mg in group A and tamsulosin 0.4mg plus placebo in group B for three days. Urethral catheter was removed three days after medical treatment and patient’s ability to void assessed at the day after catheter removal and seven days later. Patients who voided successfully were followed at least for three months. Results Mean age of patients was 59.64 ± 3.84 years in group A and 60.56 ± 4.12 years in group B (p value = 0.92). Mean prostate volume and mean residual urine were comparable between both groups (p value = 0.74 and 0.42, respectively). Fifteen patients in group A (success rate: 70%) and nineteen patients in group B (success rate: 62.7%) had failed trial without catheter (TWOC) at 7th day following AUR (p value = 0.3). No significant difference was noted between both groups regarding the rate of repeated AUR at one month and three month follow-up period (p = 0.07 and p = 0.45, respectively). Conclusion It seems that combination therapy by using 5-phosphodiesterase inhibitor and tamsulosin has no significant advantages to improve urinary retention versus tamsulosin alone. .

Genetics study and transmission electron microscopy of pili in susceptible and resistant clinical isolates of Mycobacterium tuberculosis

OBJECTIVE@#To study genetic bases and morphology of pili in Mycobacterium tuberculosis (M. tuberculosis).@*METHODS@#PCR and sequencing were used to investigate two related pili, Mtp and Flp genes in clinical isolates of M. tuberculosis. The primers were designed and PCR program were set for whole genes amplification. PCR products of the two genes from all isolates were sequenced by an applied biosystems apparatus and the results were analysed by online software. In the other hands, harvested cells from fresh cultures of isolates were undergoing specific sample preparation for sectional and negative staining for transmission electron microscopy.@*RESULTS@#Electrophoresis revealed two specific bonds of 361 bp for Mtp and 150 bp for Flp genes and confirmed primer and PCR conditions designing. There were not any mutations in sequencing results of Mtp and Flp in comparison with reference sequence. Transmission electron microscopy examination revealed two distinct types of pili in the isolates as a bundle-forming pilus and rope-like pilus. From total investigated cells, 10% harbored pili in their structure.@*CONCLUSIONS@#Two genes of pili in all clinical isolates of M. tuberculosis were conserved and two morphological types of pili were detected. We proposed that by targeting pili proteins by a suitable inhibitor, it could affect the pathogenesis especially in resistant forms.

Can procalcitonin reduce unnecessary voiding cystoureterography in children with first febrile urinary tract infection?

Recently, new predictors of vesicoureteral reflux [VUR] in children with a first febrile UTI such as Procalcitonin [PCT] were introduced as selective approaches for cystography. This study wants to show the capability of PCT in predicting presence of VUR at the first febrile UTI in children. Patients between 1 month and 15 years of age with febrile UTI were included in this prospective study. PCT values were measured through a semi-quantitative method in four grades comprising values less than 0.5, 0.5-2.0, 2.0-10.0 and above 10.0 ng/ml. The independence of PCT levels in predicting VUR were assessed after adjustment for all potential confounders using a logistic-regression model. A total of 68 patients, 54 [79.4%] girls and 14 [20.6%] boys were evaluated. PCT level demonstrated a significant difference between patients with positive VUR and those with negative VUR [P=0.012]. To calculate the independent factors that may predict the presence of VUR, all included variables were adjusted for age and sex. Results of logistic regression showed that a PCT level between 2.0 and 10.0 ng/mL could independently predict presence of VUR [Odds ratio=6.11, CI 95%= 1.22-30.77, P=0.03]. Our finding in this study showed that readily available semi-quantitative measures for PCT are feasible for detecting patients with VUR. We suggest that in semi-quantitative measurements of PCT, levels between 2.0 and 10.0 ng/ml could be an independent predictor of positive VUR

Gastrointestinal helminthes of green-winged teal (Anas crecca) from North Iran

<p><b>OBJECTIVE</b>To determine the helminth parasites of Anas crecca (A. crecca) in one of proper refuges of Iran, Fereydunkenar.</p><p><b>METHODS</b>A total number of one hundred thirty-six gastrointestinal tracts of green-winged teal (A. crecca) were collected from Fereydunkenar, Mazandaran province during September and October 2011. The gastrointestinal tracts were examined for helminth infection.</p><p><b>RESULTS</b>The total infection rate was 70.50% (96) that 68.96% (40) of males and 71.79% (56) of females shown helminthes infection. The examined A. crecca harbored one species of Nematoda, Cestoda and two species of Digenea which were as following: Contracaecum larvae (from stomach wall), Diorchis stefanskii (D. stefanskii) (from small intestine), Hypoderaeum conoideum (from small intestine) and Notocotylus attenuatus (N. attenuatus) (from caecum), respectively. There was no significant difference in the prevalence of infection between examined males and females ducks in Hypoderaeum conoideum, D. stefanskii and N. attenuatus (P>0.05) whereas a significant relationship was observed between males and females in Contracaecum larvae (P<0.05).</p><p><b>CONCLUSIONS</b>Based on the results of the present study, we conclude that A. crecca plays a prominent role in transmission of mentioned parasites. In addition, this is the first report of Contracaecum larvae, D. stefanskii and N. attenuatus from A. crecca in Iran.</p>

Genetics study and transmission electron microscopy of pili in susceptible and resistant clinical isolates of Mycobacterium tuberculosis

Objective: To study genetic bases and morphology of pili in Mycobacterium tuberculosis (M. tuberculosis). Methods: PCR and sequencing were used to investigate two related pili, Mtp and Flp genes in clinical isolates of M. tuberculosis. The primers were designed and PCR program were set for whole genes amplification. PCR products of the two genes from all isolates were sequenced by an applied biosystems apparatus and the results were analysed by online software. In the other hands, harvested cells from fresh cultures of isolates were undergoing specific sample preparation for sectional and negative staining for transmission electron microscopy. Results: Electrophoresis revealed two specific bonds of 361 bp for Mtp and 150 bp for Flp genes and confirmed primer and PCR conditions designing. There were not any mutations in sequencing results of Mtp and Flp in comparison with reference sequence. Transmission electron microscopy examination revealed two distinct types of pili in the isolates as a bundle-forming pilus and rope-like pilus. From total investigated cells, 10% harbored pili in their structure. Conclusions: Two genes of pili in all clinical isolates of M. tuberculosis were conserved and two morphological types of pili were detected. We proposed that by targeting pili proteins by a suitable inhibitor, it could affect the pathogenesis especially in resistant forms.

Development of a Disperse Dye Immunoassay Technique for Detection of Antibodies against Neospora caninum in Cattle

In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle.

Cattle fascioliasis in Gilan Province, Iran

Fasciolais a well-known parasite with a worldwide distribution and huge implications for the economic performance of domesticated herbivorous animals. It also caused deleterious effects on human health. It is prevalent in most parts of Iran, but especially in Gilan province. During 2005, 445 fecal samples taken directly from the rectum of local cattle in three different regions of Gilan and were subjected to coprological examination using saturated zinc and sodium chloride solution. The sex, age and locality of cattle were recorded. Our findings revealed an overall infection rate of 32% with the highest rate [55.2%] occurring in Talesh. No significant relationship was found between sex or age [P=0.65] and infection. Despite this, in contrast to the longheld assumption that adult cattle develops resistance to infection, our findings showed no difference between the rates of infection and the mean number of Egg Per Gram [EPG] in young and old cattle. The rate of infection in 5,820 slaughtered cattle showed that the fascioliasis is more prevalent among the animals examined in Talesh Region [P<0.01]. Meanwhile, there was a direct relationship between the amount of rainfall and the rate of infection [r=0.98, P<0.01]. The determination of Fasciola species in the liver of 70 native cattle revealed that 50 [71.4%] harbored F. gigantic, 14 [20%] had F. hepatica, and six [8.5%] had F. hepatica and F. gigantica. Therefore, F. gigantica was the dominant species in the studied areas

Production and characterization of monoclonal antibodies against brucella S [99] surface antigens

By immunizing mice with killed whole bacterial cells of Brucella abortus S [99], a panel of six hybridomas producing monoclonal antibodies [mAb] specific for the surface antigens of this bacterium were produced. ELISA was used to screen the hybridoma supernatants. Immunoblots of the cell extract indicated that three mAb were specific for S-LPS [Ba-1, Ba-2, Ba-3] and three others were reactive with major outer membrane proteins [OMP] [Ba-4, Ba-5, Ba-6]. The OMP recognized by these antibodies were the proteins with molecular masses of 25-27 kDa [Ba-4, Ba-5] and 36-38 kDa [Ba-6]. None of the four mAb including Ba-3, Ba-4, Ba-5 and Ba-6 cross reacted with any other bacteria close to Brucella abortus, but Ba-1 and Ba-2 cross reacted with B. melitensis 16M and B. suis. By using cell extract and killed whole cell Ag in ELISA, it was indicated that all mAb except Ba-6 have better reactivity with cell extract Ag, but Ba-6 mAb reacted with killed whole cell Ag better than cell extract Ag