Multilocus VNTR analysis-ompA typing of Chlamydia trachomatis isolates in Tehran, Iran.

INTRODUCTION: This study is the first to describe the genetic diversity of C. trachomatis strains derived from patients with signs and symptoms of genitourinary infections admitted to Tehran health centers and hospitals using the high-resolution genotyping method, multilocus variable-number tandem-repeat analysis with ompA sequencing (MLVA)-ompA. METHODS: One hundred and sixty-seven urogenital specimens were collected from October 2019 to July 2020. Specimens were inoculated to cell culture and examined for the presence of C. trachomatis isolates by microscopic valuation. Out of 167 samples, 19 (11.3%) viable C. trachomatis organisms were isolated in cell culture. Eighteen isolates were successfully genotyped by MLVA-ompA analysis. RESULTS: The most prevalent ompA genotypes were E, D, F and G, comprising 42%, 26.3% and 21% and 10.5% of isolates, respectively. Other genotypes were not detected from any of the samples. Out of the 18 fully genotyped isolates, 10 different MLVA-ompA genotypes were obtained. The most prevalent MLVA-ompA genotypes were 8.6.1-E (33.3%) and 8.5.2-D (16.6%). Genotype 8.6.1-E was common in both females and males. CONCLUSIONS: Our results showed that MLVA-ompA analysis was more discriminatory than ompA typing alone and, therefore, a suitable complement to ompA. Using this method, dominant genotypes in the community and transmission patterns in sexual networks could be identified. The high diversity of C. trachomatis strains in Tehran may be due to the low level of public health and awareness, and future studies are needed.

Associations between climatic parameters and the human salmonellosis in Yazd province, Iran.

Salmonella is one of the most common causes of foodborne disease outbreaks in developing countries. Climatic factors such as temperature, rainfall, and relative humidity can directly increase the growth and spread of these pathogens. Therefore, the aim of this study was to investigate long-term temporal trends and seasonal patterns of Salmonella infections as well as evaluating the effects of demographic and climatic factors on the infection incidence in Yazd province, Iran during 2012-2015. The incidence of Salmonella infections was highest among patients with the age group of ≤5 years and peaked in summer, especially during June. Contrary to expectations, no significant associations were seen between the average monthly temperature, rainfall or humidity and incidence rate (IR) of salmonellosis. Interestingly, atmospheric dust hovering was significantly associated with an increased risk of salmonellosis. Transmission pathways of Salmonella spp. in communities should be considered as a complex ecological process that animal reservoirs, socio-economic factors, and lifestyle behaviors need to be addressed in future studies.

First Detection and Characterization of Macrolide-Resistant Mycoplasma pneumoniae from People with Community-Acquired Pneumonia in Iran.

Objectives: Increasing macrolide resistance of Mycoplasma pneumoniae strains is becoming a public health concern worldwide. Nevertheless, no comprehensive genomic background of circulating isolates is available in our region. We aimed to study the genetic diversity of this microorganism using the multiple-locus variable-number tandem-repeat analysis method and to investigate the relationships between MLVA types and macrolide susceptibility profiles of the isolates. Materials and Methods: A total of 270 patients attending Tehran general university hospitals were included in this study. One throat swab was taken from each patient. M. pneumoniae was identified using culture and PCR assay. Macrolide resistance was determined using the broth microdilution method. The MLVA was performed by amplification of four variable-number tandem-repeat loci. Results: Of 270 specimens, M. pneumoniae was detected in 25.2% (n = 68) and 21.8% (n = 59) samples using PCR and culture, respectively. Approximately 56.9% of isolates were resistant to macrolides. Fifty-one of 59 M. pneumoniae isolates were divided into 6 distinct MLVA types. Conclusion: The macrolide-resistant M. pneumoniae (MRMP) rate in this study was relatively high and most of the MRMP isolates were assigned into the type 4/5/7/2. Since a significant association between MLVA type 4/5/7/2 and macrolide resistance of M. pneumoniae isolates was observed, further monitoring of genetic diversity of MRMP isolates might facilitate better understanding of epidemiology of this microorganism. Besides surveillance of the antibiotic susceptibility might be helpful to make necessary reconsiderations on guidelines for treatment of M. pneumoniae infection.

Citrobacter freundii Foodborne Disease Outbreaks Related to Environmental Conditions in Yazd Province, Iran.

BACKGROUND: The aim of this study was to assess associations of Citrobacter freundii foodborne outbreaks with environmental factors in various regions of Yazd Province, Iran, 2012-2016. METHODS: The public health surveillance data were used for one period of five years reported foodborne disease outbreaks in various regions of the Province. Multilevel regression statistical method was used to analyze associations of climatic and demographic variables with outbreaks. Significant associations were tested using likelihood ratio tests. RESULTS: Results showed a significant association between C. freundii outbreaks and air dust conditions, age groups and various regional cities. CONCLUSION: The current study revealed necessity of etiologic agent identification for use in foodborne disease outbreak guidance in future outbreaks. Systemic surveillance schemes can help prevent and control similar scenarios using reports of environmental effects on foodborne disease outbreaks.

Molecular epidemiology and genetic characterization of Shigella in pediatric patients in Iran.

Infection with Shigella is considered a major cause of morbidity and mortality in children with diarrhea in developing countries, especially in Iran. Due to the importance of country-level epidemiological data, molecular characterization of genetic determinants of Shigella spp. is a necessity. The aim of the present study was to investigate the prevalence of integron types, bla-CTX-M, bla-SHV and blaTEM ß-lactamase genes of Shigella isolates in pediatric patients in Tehran, Iran. In a time period of 18 months from May 2015 to August 2017, 75 Shigella spp. were isolated from non-duplicative diarrheal stool specimens in six different hospitals in Tehran. The isolates from patients were further analyzed for their antibiotic susceptibility and extended-spectrum beta-lactamase (ESBL) production. Polymerase chain reaction was performed for amplification of the integrons (I, II, III), TEM, SHV, CTX-M15. The prevalence of S. sonnei, S. flexneri, S. dysenteriae and S. boydii were 40 (53.3%), 33 (44%), 1 (1.3%) and 1 (1.3%), respectively. The results of an antimicrobial resistance test showed that the high percentage of resistance to nalidixic acid (NA), ampicillin (AMP) and trimethoprim/sulfamethoxazole (SXT) included 38 (50.6%), 59 (81.3%) and 64 (88%) isolates, respectively. Further results revealed that 52% and 76% of Shigella isolates carried intI and intII genes, respectively. In this study, the rates of CTX-M (10.7%), SHV (28%) and TEM (21.3%) were determined, all of which were positive for blaCTX-M15. This study showed the high prevalence of multidrug resistant S. sonnei and S. flexneri. Furthermore, it highlighted the increasing integrons (intI and intII) and ESBL genes, especially blaCTX-M15, in Shigella isolates.

High frequency of Mycoplasma pneumoniae among patients with atypical pneumonia in Tehran, Iran.

INTRODUCTION: Mycoplasma pneumoniae is a major cause of atypical community-acquired pneumonia (CAP) with a prevalence range of 15-20% and up to 40% in adults and children, respectively. In Iran, the recorded frequency ranges between 1-6.15%. We aimed to investigate the frequency of M. pneumoniae among patients with atypical pneumonia acquired from the community. METHODS: Over a period of 5 months between January and June 2017, 520 patients with suspected CAP, who had been to the hospital outpatient clinics of Tehran University, were enrolled in this study. Throat swab specimens were obtained from 110 outpatients who presented with symptoms of atypical pneumonia. M. pneumoniae was identified via culture and biochemical tests, such as fermentation of glucose and arginine, hemolysis, and hemadsorption. For confirmation, PCR was performed to amplify the gene fragment coding for p1 adhesin. RESULTS: The major and minor clinical signs of the patients were dyspnea (67.3%) and nausea (15.5%), respectively. Out of 110 specimens, 25 (22.7%) and 29 (26.4%) isolates were identified to be M. pneumoniae via culture and molecular assay, respectively. Comparing the results of the two methods, the PCR showed better sensitivity and rapidity for the detection of M. pneumoniae. There was a high congruence between culture and the PCR assay; kappa level was 'almost perfect' (κ=0.90). CONCLUSION: This is the first report of high frequency of M. pneumoniae in our region. This finding can serve as baseline information for further investigation and confirmation of the potential epidemics of M. pneumoniae pneumonia in our community.

Prevalence of virulence genes and their association with biofilm formation in VRE faecium isolates from Ahvaz, Iran.

INTRODUCTION: Vancomycin-resistant Enterococcus faecium (VREfm) is a common cause of nosocomial infections. Biofilm formation is an important factor in recurrence of infections, facilitating transfer of genetic elements, leading to treatment failures. The aim of this study was to investigate the virulence genes in biofilm producing isolates and to determine possible association between biofilm formation and the presence of these genes; also to determine association between antibiotic susceptibility patterns of VREfm isolates and their biofilm formation ability. METHODOLOGY: A total of 57 isolates of VREfm were recovered from different sources of hospitals under Ahvaz University, Iran. The isolates were examined by conventional microbiological methods and molecular test using PCR. The antibiotic susceptibility patterns of the isolates were determined by disk-diffusion and E-test. The biofilm formation ability of the isolates was investigated by Modified Congo red agar and microtiter plate techniques. The presence of virulence genes was examined using Multiplex-PCR method. RESULTS: Out of 57 VREfm isolates, 63.15% of isolates were biofilm producers. The frequency of biofilm producing isolates from clinical specimens, colonized patients and environmental sources were 78.26%, 60%, and 42.85%, respectively. The prevalence of acm, esp and hyl genes among biofilm producing isolates was 86.10%, 55.56% and 52.77%, respectively. There was statistically significant association between esp gene and biofilm formation among isolates from the clinical specimens. CONCLUSION: Clinical isolates producing biofilms showed a positive association with the presence of the esp. Our study further suggests that the link between virulence genes and biofilms is affected by the environmental context.

CbpM and CbpG of Streptococcus Pneumoniae Elicit a High Protection in Mice Challenged with a Serotype 19F Pneumococcus.

Among many pneumococcal antigens, choline-binding proteins (CPBs) display a high immunogenicity in animal models. This study aims to determine the immunogenicity of CbpM, CbpG and CbpL proteins of Streptococcus pneumoniae in a mice model. The genes were cloned into pET21a expression vector and the recombinant proteins were produced. Mice were immunized with the purified recombinant proteins. Subsequently, the mice were challenged with S. pneumoniae ATCC 49619 (2×106 CFU) and their survival and bacterial clearances were followed 24 hours after infection. The antibody responses of the mice were determined by ELISA assay. The opsonophagocytosis assay was performed using rabbit's sera. Passive immunization was carried out using two doses of anti-CbPs antibodies. Finally, these mice were  experimentally infected with virulent bacteria and the protective effects of two doses of 10 and 100 µg/mL by monitoring the survival rate and bacterial clearance were determined at 2, 3 and 7 days after bacterial challenge. The mice actively immunized with CbpM, CbpG and CbpL recombinant proteins showed survival rate of 100%, 85% and 75%, respectively. The survival rates among passively immunized mice groups which received 100 µg/mL dose of anti-CbpM, anti-CbpG and anti- CbpL were 50%, 50% and 25%, respectively. The rates of opsonization with rabbit's antibodies against CbpM, CbpG, and CbpL  at 100 µg/mL doses was 45.6%, 14.7% and 82.3%, and at 10 µg/mL was 12.9%, 12.2% and 9.35%, respectively. Our findings suggest that the recombinant proteins particularly CbpM and CbpG can protect the mice against pneumococcus19F serotype and effectively induce a protective antibody response. Thus, CbpG and CbpM proteins might be used as suitable vaccine candidate in pneumococcal vaccine formulations.

Biofilm formation and antimicrobial resistance in methicillin-resistant Staphylococcus aureus isolated from burn patients, Iran.

INTRODUCTION: Burns are the most serious forms of trauma and a major cause of mortality worldwide. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common pathogens of burn wound infections; treatment has faced serious problems due to antibiotic resistance in these strains. Biofilm formation, which increases antibiotic resistance capabilities and is considered to be a virulence factor, also causes treatment failure and recurrent staphylococcal infections in burn patients. METHODOLOGY: A total of 135 pus/wound swabs were collected; S. aureus was identified by confirmatory tests. The icaA/D and mecA genes were detected in DNA extracts by polymerase chain reaction assay separately. To determine the prevalence of biofilm formation, a modified Congo red agar and the microtiter plate method were used. Investigation of antibiotic resistance was performed using the disk diffusion method. RESULTS: S. aureus (48.87%) was identified in 65 (48.87%) samples, of which 40 (61.53%) were confirmed to be MRSA. Among MRSA and methicillin-sensitive S. aureus (MSSA) isolates, 97.5% and 60% produced biofilm, respectively. Resistance of MRSA isolates to amikacin, ceftriaxone, ciprofloxacin, erythromycin, gentamicin, mupirocin, rifampin, tetracycline, and tobramycin was 64.1%, 76.92%, 51.28%, 87.18%, 71.8%, 10.26%, 5.13%, 89.74%, and 61.54%, respectively. All MRSA and MSSA isolates were susceptible to fusidic acid, linezolid, teicoplanin, tigecycline, and vancomycin. CONCLUSIONS: The high prevalence of biofilm-producing, drug-resistant S. aureus isolates in our study suggests that epidemiological studies on the characteristics of common strains found in burn centers and a definition of their antibiotic resistance pattern would be helpful for therapeutic decisions.