فهرست مطالب

Iranian Journal of Microbiology
Volume:14 Issue: 5, Oct 2022

  • تاریخ انتشار: 1401/08/18
  • تعداد عناوین: 20
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  • Partha Satpathi, Sayantani Endow, Kripasindhu Gantait, Pearl Chand, Abhishek Jaiswal, Biswajit Jana, Panchanan Kundu, Tarapada Ghosh Pages 611-616
    Background and Objectives

    Vaccination being the only way to reduce mortality from the dreaded COVID-19 disease, the vaccine was introduced in India as per the advice of the National Expert Group on January 16, 2021. Duration of immune response elicited by the vaccines has always been a matter of content. With new variants emerging every other day, the study was done to look for the antibody response in vaccine recipients post second dose of vaccination.

    Materials and Methods

    A longitudinal observational study was conducted from August 2021 to February 2022 in fully vaccinated individuals who took either Covishield (AZD1222) or Covaxin (BBV-152). Blood was collected from the individuals at 12-16 weeks post-vaccination to look for IgG antibody response against S1 spike protein of SARS-CoV2 by ELISA. Follow-up was done at 32 weeks post the second dose in individuals who had received Covishield.

    Results

    Among 176 individuals, IgG antibody against S1 spike protein was found to be positive in 89.7% (158). Covishield recipients showed higher antibody response (99.1%) as compared to Covaxin recipients (71%). Antibody response was higher in males, individuals less than 50 years, and non-comorbid individuals. Of 38 Covishield recipients, IgG antibody response was positive in 28 (73.6%) individuals when followed up at 32 weeks post the second vaccination dose.

    Conclusion

    The study gives us input with regard to the long-term antibody kinetics of both vaccines. The study has a follow-up plan to co-relate the antibody response to the neutralization test.

    Keywords: Immunoglobulin G, Vaccination, India, Spike glycoprotein
  • Farzad Ziarani, Alireza Tahamtan, Hasan Safari, Alijan Tabarraei, Yousef Shahamat Pages 617-623
    Background and Objectives

    Coronavirus disease 2019 (COVID-19) is a pandemic caused by the novel virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Knowing the virus's behavior and its persistence in different environments are crucial and will lead to the proper management of the disease. In this study, air, surface, and sewage samples were taken from different parts of referral hospitals for COVID-19.

    Materials and Methods

    Air samples were taken with impinger, surface samples with swabs, and sewage samples were taken from the hospital wastewater treatment plant. After viral genome extraction, a real-time RT-PCR test was applied to confirm the presence of SARS-CoV-2 RNA in the collected samples.

    Results

    The virus genome could be traced in the wards and wastewater related to hospitalized COVID-19 patients. Overally, 29%, 16%, and 37.5% of air, surface, and sewage samples were positive for the SARS-CoV-2 genome, respectively.

    Conclusion

    Findings of such studies provide valuable results regarding the degree of contamination of hospital environments and the risk of virus transmission in different environments and among hospital staff and patients.

    Keywords: COVID-19, SARS-CoV-2, Hospital contamination, Air pollution, Environmental contamination, Wastewater
  • Hamid Morovati, Saba Eslami, Hesam Farzaneh Bonab, Mohammad Kord, Sima Darabian Pages 624-635
    Background and Objectives

    There is a poor understanding about the prevalence and characteristics of secondary bacterial and fungal infections among Coronavirus diseases 2019 (COVID-19) superinfection in hospitalized patients.

    Materials and Methods

    Four hundred COVID-19-proven patients were enrolled in this study. Nasal swabs for molecular assay (Real-time PCR) and sputum samples for further microbiological assays were collected. Following a broad-spectrum search, a meta-analysis was performed using StatsDirect software (version 2.7.9) according to the DerSimonian and Laird method applying the random-effects models.

    Results

    Streptococcus spp. (21.5%) and Staphylococcus spp. (16.7%) had the highest prevalence of bacterial coinfection among the COVID-19 patients, while Acinetobacter spp. had the lowest prevalence (4.2%). Among fungal coinfections, Candida albicans was the most prevalent (6.7%), and Aspergillus spp. was the lowest (2%). Males, elderly patients, patients with a history of underlying diseases and drug use, patients who showed acute clinical symptoms, and patients with a prolonged hospital stay had a higher incidence of secondary infections (P-value <0.05). The pooled prevalence for bacterial and fungal coinfections was 33.52% (95% CI: 18.12 to 50.98; I2: 99.4%; P-value: <0.0001).

    Conclusion

    We suggest designing additional research with a larger target population and diagnostic molecular analyses to depict a more realistic view of the coinfection status.

    Keywords: COVID-19, Secondary infection, Nosocomial infection
  • Raunak Bir, Sarita Mohapatra, Arvind Kumar, Nazneen Arif, Sonu Tyagi, Aswin AK, Hitender Gautam, Seema Sood, Bimal Das, Arti Kapil Pages 636-644
    Background and Objectives

    Urinary tract infection is one of the most common bacterial infections causing high morbidity and mortality. The alarming rise of multidrug-resistant uropathogens worldwide forced the clinician to rethink the old drugs like Fosfomycin for its therapeutic management. Our objective was to compare agar dilution, disc diffusion and E-test method for antimicrobial susceptibility testing of Fosfomycin against different drug-resistant uropathogens.

    Materials and Methods

    Consecutive 181 uropathogens were tested for Fosfomycin susceptibility using agar dilution, disc diffusion and E-test. Results were interpreted using Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints. Whole genome sequencing analysis was done on the 4 XDR/PDR Fosfomycin resistant Klebsiella pneumoniae isolates.

    Results

    Escherichia coli was found as the most common (62.4%) uropathogen followed by Klebsiella pneumoniae (21%). Considering agar dilution as the gold standard, 6.1% of isolates were resistant to Fosfomycin. Following CLSI breakpoints, the susceptibility of Escherichia coli, Klebsiella pneumoniae, other Enterobacterales and Pseudomonas aeruginosa were 92.9%, 92.1%, 100%, 100%; whereas using EUCAST breakpoints the susceptibility rates were 85.7%, 86.9%, 92.9%, and 100%, respectively. The essential agreement, categorical agreement, major error, and very major error for E-test/ disc diffusion for all the organisms were 91.2%/Not Applicable, 95%/93.9%, 1.8%/4.7%, 9.1%/9.1%, respectively. Whole-genome sequencing showed mutation UhpT gene as well as the presence of plasmid-mediated fosA5 or fosA6 genes conferring Fosfomycin resistance.

    Conclusion

    This result supports very low resistance of Enterobacterales against Fosfomycin; hence should be considered a valuable option to treat multidrug-resistant uropathogens. Disc diffusion was observed to be a convenient method for Fosfomycin susceptibility testing compared to agar dilution.

    Keywords: Fosfomycin, Agar dilution, Disc diffusion, E-test, Enterobacterales
  • Zeinab Ibraheem Sayed, Mohamed Fawzy Abdel-Ghany, Shabaan Hashem Ahmed, Amany Mohmed Adawy, Rawhia Abd El–Hamid Pages 645-652
    Background and Objectives

    Streptococcus pseudopneumoniae is a member of viridans streptococci. It is non-capsulated, bile insoluble and optochin susceptible in ambient air but resistant in 5% CO2. This study aimed to isolate S. pseudopneumoniae from sputum specimens of patients admitted to Chest Department and Chest ICU of Assiut University hospitals, differentiate it from Streptococcus pneumoniae in addition, to evaluate the prevalence of Streptococcus pseudopneumoniae in clinical isolates by phenotypic and genotypic methods, to subject the isolates to antimicrobial susceptibility testing using agar disc diffusion method.

    Materials and Methods

    Isolation of Streptococcus pseudopneumoniae from sputum sample and doing phenotypic test (optochin susceptibility test,bile susceptibility test and antimicrobial susceptibility test) and genotypic test by polymerase chain reaction (PCR) for five genes: CpsA, LytA, AliB-like ORF2, 16S rRNA and Spn9802 genes.

    Results

    Twenty isolates of S. pseudopneumoniae were diagnosed phenotypically by optochin susceptibility and bile solubility tests followed by genotypic characterization by polymerase chain reaction (PCR) for five genes: CpsA, LytA, AliB-like ORF2, 16S rRNA and Spn9802 genes. The prevalence of S. pseudopneumoniae among studied patients was 10% (20/200).

    Conclusion

    The pure growth of S. pseudopneumoniae from sputum samples together with the great percentage of antibiotic resistance should raise attention to the clinical importance of this organism.

    Keywords: Streptococcus, Respiratory tract diseases, Polymerase chain reaction
  • Bui Son, Tang Hai, Tran Cuong, Duong Chinh, Thi-Hong-Hanh Le, Nguyen Dung, Vu Dinh, Do Anh Pages 653-661
    Background and Objectives

    Identification of pnemococcal serotypes and antimicrobial resistance provides helpful information for the use of suitable vaccines and antibiotics; however, very limited data is available on these issues in Vietnam. The present study aimed to find the serotype distribution and drug resistance patterns of Streptococcus pneumoniae isolated from unvaccinated children less than 5 years of age with pneumonia at a province in centre Vietnam.

    Materials and Methods

    A total of 126 clinical pnemococcal strains isolated from unvaccinated children less than 5 years of age with pneumonia at the Nghe An province, Vietnam between Nov 2019 and Mar 2021. All strains were identified using conventional microbiological method, VITEK® 2 Compact system, specific PCR and sequencing. The serotypes and antimicrobial resistance patterns of pnemococcal strains were determined using the multiplex PCR assays and VITEK® 2 Compact system.

    Results

    The results showed that, eight different pneumococcal serotypes were identified. The most common serotypes were 19F (67.46%), followed by 23F (10.32%), 19A (9.52%), 6A/B (3.17%), 15A (2.38%), 9V (3.17%), 11A (1.59%) and 14 (0.80%), respectively. More than half of the pneumococcal strains were non-susceptible to penicillin. The resistance rate to ceftriaxone and cefotaxime were 41.3% and 50.8%. The percentage of pneumococci strains resistant to clarithromycin, azithromycin, erythromycin, cotrimoxazole, tetracyclin, and clindamycin were more than 93% of all strains. All pneumococcal serotypes were highly resistant to clarithromycin, azithromycin, erythromycin, cotrimoxazole, and clindamycin.

    Conclusion

    Our findings showed high antibiotic resistance rates of the strains causing pneumococcal pneumonia, mostly macrolide resistance, among unvaccinated children.

    Keywords: Streptococcus pneumoniae, Serotypes, Antibiotic resistance, Children, Pneumonia
  • Fatima Kaleem, Irum Aftab, Umme Farwa, Hassan Saleem, Saima Ishtiaq, Saima Syed, Shahid Ahmad Abbasi Pages 662-668
    Background and Objectives

    This study was designed to determine the in vitro efficacy of mecillinam against extended spectrum beta lactamse producing Enterobacterales.

    Materials and Methods

    After proper permission from Ethical Review Committee of the Institute, all samples yielding growth of ESBL producing Enterobacterales were part of the study and were processed according to routine microbiological procedures. Routine antibiotic sensitivity testing was done on Muller Hinton Agar by Modified Kirby Bauer Method. All Gram negative isolates were subjected to concomitant detection of ESBL production by double disc synergy method. All ESBL producers were then subjected to the mecillinam Minimum Inhibitory Concentration (MIC) determination by E test. The results were interpreted as per CLSI Guidelines.

    Results

    A total of 120 ESBL producing Enterobacterales isolates were included in the study. The mean age of patients with ESBL infection was 45 ± 18.7 years. There were 44% male and 55% female patients. Majority of the ESBL producing Enterobacterales were isolated from urine samples (56%), followed by pus. Among the isolated organisms, Escherichia coli (45%) was the most frequently isolated organism followed by Klebsiella spp. (22%). Overall 83% of the isolates turned out to be sensitive to mecillinam. MIC50 of mecillinam against ESBL producing Gram negative rods (GNR) turned out to be 1 ug/ml and MIC90 turned out to be 2 ug/ml.

    Conclusion

    Mecillinam shows good in vitro efficacy against ESBL producing Enterobacterales in our study. Further studies with more sample size and from diverse areas across the country should be done to evaluate its efficacy.

    Keywords: Extended spectrum β-lactamase, In vitro susceptibility
  • Lahbib Hibaoui, Abdelhamid Massik, Zina Lebbar, Ghita Yahyaoui, Mustapha Mahmoud, Laila Bougnouch, Naoual Hamass, Laila Chbani, Bahia Bennani, Mohamed Amine Berrahou, Mounia Lakhdar Idriss, Moustapha Hida Pages 669-676
    Background and Objectives

    Infesting nearly 50% of the world's population, Helicobacter pylori are thought to cause peptic ulcers, as well as gastric adenocarcinoma. Several diagnostic methods are available to detect this bacterium; however, at least two must be used together for an accurate diagnosis. This study evaluated the use of rapid urease test for diagnosis of H. pylori infection in a pediatric population.

    Materials and Methods

    Five gastric biopsies were taken from children during a 2-year period for the purpose of histological, molecular, bacteriological culture, and rapid urease testing.

    Results

    Among 83 children, 38 were male, and 45 were female with an age ranging of 2 to 15 years. The infected group represented 31%. The rapid urease test had a sensitivity of 88.5%, a negative predictive value of 94%, a specificity of 84.2%, and a positive predictive value of 72%.

    Conclusion

    A rapid urease test may be appropriate for ruling out H. pylori infection after a negative result. The positive results however, may be confirmed by a second invasive test.

    Keywords: Helicobacter pylori, Children, Invasive tests, Rapid urease test, Polymerase chain reaction, Histology, Biopsyculture
  • Siamak Mohebi, Roghieh Saboorian, Saeed Shams Pages 677-682
    Background and Objectives

    Vibrio fluvialis is a Gram-negative, bacillus-shaped, curved bacterium known as an emerging pathogen. There are reports of outbreaks caused by this bacterium worldwide. Iran, especially Qom province, is an endemic region for gastrointestinal diseases caused by Vibrio species. So, the aim was to isolate V. fluvialis from clinical and environmental samples.

    Materials and Methods

    During six months, 363 clinical and surface water samples were evaluated. The samples were cultured on specific media, and all incubated for 24 hours at 37°C. Suspicious colonies were evaluated by Gram staining and biochemical tests. The BD Phoenix automated microbiology system was used for the final confirmation of the isolated bacteria. Evaluation of antibiotic resistance of isolated strains was also performed according to CLSI standard.

    Results

    Eight cases (2.2%) of V. fluvialis, including seven from surface water samples (87.5%) and one from clinical samples (12.5%), were isolated. Based on antimicrobial susceptibility testing, all V. fluvialis isolates were susceptible to amikacin, gentamicin, trimethoprim/sulfamethoxazole, ciprofloxacin, tetracycline, ceftazidime, and chloramphenicol. High-level resistance to ampicillin and amoxicillin/clavulanate was also observed. V. fluvialis-infected patient had a mild fever, watery diarrhea, vomiting, nausea, and abdominal cramps that were manifested after drinking contaminated water or eating contaminated vegetables. The patient's symptoms recovered without antibiotic therapy after four days, resulting in self-limiting disease.

    Conclusion

    The current study is the first human case of V. fluvialis infection isolated in Iran. Therefore, monitoring of water and food samples should be done routinely.

    Keywords: Vibrio fluvialis, Gastroenteritis, Drug resistance, Microbial, Iran
  • Nasrin Hamidian, Saeed Salari, Ahmad Rashki, Mohsen Najimi Pages 683-688
    Background and Objectives

    Microbial safety of the fresh fruits is a global concern. The production of strawberry (Fragaria ananassa) in Iran has grown 4.5% from 2016 to 2019. Little information is available about the microbiological quality of strawberry produced in Iran. The objective of this investigation was to assess the fecal Escherichia coli (FEC) contamination of fresh strawberries of south of Kerman province where is the centers of production and supply of strawberries to other parts of Iran.

    Materials and Methods

    In this cross-sectional descriptive study, the FEC of a total number of 109 strawberry samples, which were collected from green-houses of strawberry of Kerman province during three months, were enumerated using the most probable number (MPN) as described by Iranian National Standards Organization, protocol No.2946, and interpreted using the latest statute released by Iran Veterinary Organization, Executive order number 2946, released in 2011.

    Results

    MPN of FEC counted in strawberries ranged between <0.3 (n=37) and >110 (n=19) per gram (g) having a mean, mode, and median value of 250.3, <0.3, and 9.4 MPN/g, respectively. More than one-fourth of the samples (28.44%) were polluted with FEC at a level of >100 MPN/g.

    Conclusion

    Our findings may be resulted from the sanitary quality of the farm and strawberries of the study area, which indicated that the microbial safety of the strawberries in this survey was not satisfactory, alarming public health.

    Keywords: Contamination, Escherichia coli, Fragaria, Fruit, Iran, Public health
  • Farzaneh Nazari, Parvaneh Jafari, Bizhan Nomanpour, Kambiz Varmira, Farshid Raissi Pages 689-697
    Background and Objectives

    Breast cancer is the second leading cause of death and one of the most common malignancies among women in the world. The aim of this study was to investigate the preventive effects of postbiotic consisting of sonicated Bifidobacterium bifidum cells on triple negative breast cancer.

    Materials and Methods

    Thirty-six female BALB/c mice aged 5-7 weeks were randomly divided into 3 groups (n=12): Ctrl-, healthy mice; Ctrl+, mice with breast cancer with no treatment; and Postbiotic, mice with orally gavage postbiotic before and after 4T1 cell line transplantation. Cancer progress and the effects of postbiotic were assayed by histological, immunohistochemical and gene expression quantification.

    Results

    The histological results showed that administration postbiotic consisting of B. bifidum significantly decreased carcinogenesis in terms of tumor incidence, multiplicity and volume. The tumor progress was suppressed by oral intake of B. bifidum as showed by p53 and Ki-67 expression. Furthermore, Oral intake of postbiotic resulted in extended survival of mice and inhibited sever weigh loss.

    Conclusion

    Pretreatment with sonication killed B. bifidum, as a postbiotic, inhibited breast cancer progress and malignancy.

    Keywords: Triple negative breast neoplasm, Bifidobacterium bifidum, Ki67 antigen, Tumor suppressor protein p53
  • Fatemeh Mehdizadeh Sari, Shiva Mirkalantari *, Shohreh Nikoo, Fatemeh Sepahvand, Leila Allahqoli, Arezoo Asadi, Malihe Talebi Pages 698-704
    Background and Objectives

    Endometriosis is defined as the presence of endometrial tissue outside the uterine cavity. Peripheral blood monocytes cells (PBMCs) may have altered function to some extent in women with endometriosis. Lactobacillus acidophilus is a probiotic bacterium within the human body with the ability of alleviating many inflammatory diseases. Here, we examined the effect of L. acidophilus on PBMCs of endometriosis patients.

    Materials and Methods

    In this study, peripheral blood samples were obtained from endometriosis patients (n=11) and non-endometriosis individuals (n=11). After isolation of peripheral blood mononuclear cells with Ficoll, cells were cultured in the presence and absence of phytohemagglutinin. Also, these cells were co-cultured with 1×106 CFU/ml of L. acidophilus. IL-6 and IL-1 cytokines were measured by ELISA method and the two groups were evaluated and compared.

    Results

    The results showed that in endometriosis patients, the production of pro-inflammatory cytokines, including IL-1 and IL-6, by PBMC was increased compared to non-endometriosis subjects, and stimuli such as PHA intensified this elevation. Also, L. acidophilus increased the levels of pro-inflammatory cytokines including IL-1 and IL-6. However, the production of these cytokines decreased due to the modulatory properties of bacterial cells after 48 h.

    Conclusion

     According to the results of the current study, IL-1 and IL-6 production was significantly increased in PMBCs of endometriosis patients compared to that of the healthy controls. Also, Lactobacillus acidophilus was considered as an antigenic compound and in duced IL-1 and IL-6 production. According to these results, probiotics can be further used for the treatment of endometriosis patients and more investigations are needed to con firm these results.

    Keywords: Probiotic, Endometriosis, Lactibacillus acidophilus, Inflammatory cytokines
  • Keyvan Esmaeili-Fard-Barzegar, Shahin Najar-Peerayeh, Ramin Mazaheri Nezhad Fard, Bita Bakhshi Pages 705-711
    Background and Objectives

    Acinetobacter baumannii (A. baumannii), an opportunistic pathogen, has been isolated from sewage, soil, and hospital wards. The prevalence of multidrug-resistance A. baumannii has seriously caused a health crisis in hospital settings. Bacteriophages have been used as an alternative therapy for control carbapenem-resistant bacteria-caused infections. We aimed to assay lytic effect of a filamentous phage on clinical bacterial strains.

    Materials and Methods

    500-ml water samples was collected from sewage in Tehran. Sewage samples were precipitated at 6000 rpm for 10 minutes and filtered using 0.45-µm syringe filters. Bacteriophage was isolated using double-layer agar assay and evaluated its stability at various pH and temperature ranges. In addition, the stability of the phage was assayed at chloroform 0.1%.

    Results

    Transmission electron microscopy imagine showed phage is filamentous called vB-AbaI-TMU2. The phage affected on its own host so that could not effect on any Pseudomonas aeruginosa and Klebsiella pneumoniae strains. vB-AbaI-TMU2 phage was stable at pH 5 and 7 and also temperatures 25 and 37°C. vB-AbaI-TMU2 was stable at chloroform 0.1%. vB-AbaI-TMU2 phage infected carbapenem-resistant A. baumannii strains while other bacterial strains were resistant to.

    Conclusion

    The present study indicated the isolated phage had a narrow host range and is susceptible to various pH values and temperatures.

    Keywords: Bacteriophage, Acinetobacter baumannii, Sewage, Drug resistance, Inoviridae
  • Zeinab Rezaei, Ameneh Elikaei, Seyed Mahmoud Barzi, Morvarid Shafiei Pages 712-720
    Background and Objectives

    Phage therapy has gained interest as an alternative treatment for methicillin-resistant Staphylococcus aureus (MRSA) infections. The purpose of this study was to isolate and characterize an effective bacteriophage against isolates of MRSA.

    Materials and Methods

    Bacteriophage was isolated from hospital sewage. Lytic activity and the titers of phage lysates were measured using spot test and double-layer plaque assay. The phage characterization was determined through transmission electron microscopy. Adsorption rate, host range and stability tests were investigated. The latent period and burst size were estimated from a one-step growth curve. The effect of bacteriophage against MRSA biofilms was determined and Real-time PCR was used to assess the effects of the bacteriophage on the expression of the biofilm-associated genes.

    Results

    TEM results showed that the phage resembled the Cystoviridae family. Its latent period was 30 min, corresponding to about 71/43 phage particles per infected cell. The phage had a broad host range and it was most stable at 37°C and pH 7. It was sensitive to NaCl concentrations. The expressions of the biofilm-associated genes were significantly reduced in the presence of the phage.

    Conclusion

    The isolated phage was effective against MRSA strains and it can be an optional strategy of controlling biofilm development.

    Keywords: Bacteriophage, Methicillin-resistant Staphylococcus aureus, Phage therapy, Wounds
  • Nasim Rouhizohrab, Fatemeh Mohammadipanah Pages 721-729
    Background and Objectives

    Myxobacteria initially recognized by their complex life cycle and social behavior are progressively explored for their bioactive secondary metabolites. However, isolation of myxobacteria usually is accompanied by bacterial and fungal contaminations due to the direct cultivation of soil on isolation media, which results in severe challenges in the purification of myxobacteria. Therefore, it is necessary to improve their purification techniques from natural samples to the discovery of new biomolecules.

    Materials and Methods

    In the present study, six physichochemical methods were assessed for their efficacy in the purification of myxobacterial strains and specially from contaminants of Microvirga spp.

    Results

    Among the evaluated treatments, purification of fruiting bodies using a combination of ultrasonication and heat treatment was identified as the effective protocol with 80% success rate in the purification of myxobacterial strains and reducing up to 90% of the contaminating bacteria.

    Conclusion

    Concerning the problematic contamination of myxobacterial isolates, the introduced approach can retrieve the myxobacterial strains which are often suppressed by the over growth of contaminations especially root symbiotic bacteria namely Microvirga spp.

    Keywords: Myxobacteria, Strain purification, Heat treatment, Ultrasonication
  • Dilip Karad, Ravi Tandon, Ajay Arya, Kailash Sonawane, Anusaya Chavan, Arun Kharat Pages 730-739
    Background and Objectives

    Even after four decades, HIV infection remains a global challenge and a leading cause of mortality in adults across the world. Anti-retroviral therapy (ART) that controls HIV viremia, is now available through public health facilities in India but drug resistance, which is likely to develop among these individuals remains poorly studied in India. The objectives of present study are to find out the HIV-1 virus subtypes, drug resistance mutations and HIV-1 drug resistance to NRTI, NNRTI and protease inhibitors in the Solapur district, India.

    Materials and Methods

    In a cross sectional study, forty two ART-experienced HIV-1-infected patients with CD4+ count < 200 cells ml-1 and viral load (VL) > 3, 000 copies ml-1 were recruited. All patients belonged to Maharashtra State of India near Barshi Solapur and had been on ART treatment for over 5 years. EDTA whole blood from HIV-1-infected patients was centrifuged and the viral nucleic acid was purified from the plasma. Viral nucleic acid was amplified by PCR using protease and reverse transcriptase specific primers. The resulting amplicons were sequenced and studied for mutations. The tools from Stanford University website were used for subtyping of HIV-1 and identification of mutations conferring drug resistance.

    Results

    In present investigation, HIV-1 subtypes were subtype C in 37 (88.09%), subtype CRF01_AE in 2 (4.76%), and subtype A in 3 patients (7.14%). Drug resistance mutations of NRTI, NNRTI and protease were observed in 15 (37.71%) of 42 patients tested. Drug resistance for NRTI was observed in 12 (28.57%) and for NNRTI in 13 (30.95%) patients. No drug resistance was observed for protease inhibitors.

    Conclusion

    Considerable HIV-1 drug resistance exists among patients receiving ART from a rural areas of India, suggesting more studies from rural region are required to prevent development of resistance to ART.

    Keywords: HIV-1, Antiretroviral therapy, CD4+ count, Viral load, Drug resistance
  • Anitha Madhavan, Arun Sachu, Abel Samuel, Jayalakshmi Vasudevapanicker Pages 740-745
    Background and Objectives

    Cryptococcosis is an opportunistic mycosis, caused by Cryptococcus neoformans. Cryptococcal meningitis is one of the most fatal opportunistic infections associated with human immunodeficiency virus (HIV) infection. The aim of this study was to find the prevalence of cryptococcal antigenemia in people living with HIV (PLHA) and also to find the prevalence of opportunistic infections among these patients.

    Materials and Methods

    A total of 204 non duplicate samples were collected from people with HIV aged above 18 years. Samples with CD4 count less than 300 were included in the study. Cryptococcal antigen detection was done by CrAg Lateral flow assay.

    Results

    None of the patients in our study were positive for cryptococcal antigen. Opportunistic infections were observed in 82 (40.2%) HIV positive patients. Candidiasis, tuberculosis and Pneumocystis jiroveci pneumonia were the most common opportunistic infections.

    Conclusion

    This is the first study from the southern state of Kerala on the prevalence of Cryptococcal antigenemia among HIV positive individuals. The study showed that routine screening for cryptococcal antigen will not be cost effective in our population. Similar to other studies, eventhough candidiasis, tuberculosis and PCP were more commonly seen among people with CD4 count < 200 cells/mm3, there was no statistically significant association.

    Keywords: Cryptococcosis, Toxoplasmosis, Opportunistic infections, Tuberculosis, Candidiasis
  • Nahid Omidi, Azarakhsh Azaran, Manoochehr Makvandi, Gholamreza Khataminia, Kambiz Ahmadi Angali, Shahram Jalilian Pages 746-758
    Background and Objectives

    Adenovirus species B, C, D, and E are the most common causes of ocular manifestations caused by adenoviruses. FDA-approved treatment agents for adenovirus infections are not available. Cell-mediated immunity is the major protective mechanism versus human adenoviruses (HAdVs) infection and T cells specific for peptide epitopes from nonstructural proteins can prevent adenoviral dissemination. E1A CR2 region of HAdVs Epitopes predicted for reinforcing cytotoxic T lymphocytes (CTLs) in the EKC patients. Among human adenoviruses E1 protein, four distinct E1A regions had a significantly higher level of homology than the rest of E1A protein. E1A protein inhibits IFN signal transduction. Epitope-based vaccines are designed to have flexible and simple methods to synthesize a vaccine, using an adjuvant to trigger fast immune responses. CTL epitopes were applied to create a multiepitope vaccine. Conserve region1 (CR1) and CR3 have less antigenicity compared to CR2. Additionally, CR3 in HAdV-D8 contains three toxic areas. CR4 similar to the two regions CR1 and CR3 do not show acceptable antigenic properties.

    Materials and Methods

    Bioinformatics’ tools were used to predict, refine and validate the 3D structure of the construct. Effective binding was predicted by protein-protein docking of the epitope vaccine with MHC-I molecules and revealed the safety and efficacy of the predicted vaccine construct.

    Results

    In silico analysis show that rising levels of cytotoxic CD8 + T cells, TH1 cells, macrophages, and neutrophils are linked to IFN-dominant TH1-type responses, which are detected in putative immune individuals.

    Conclusion

    Combined with 3D protein modeling, this study predicted the epitopes of E1A CR2 protein in HAdVs.

    Keywords: In silico model, Adenovirus E1A proteins, Keratoconjunctivitis, Molecular docking, Cytotoxic T-lymphocyte
  • Roya Pirmoradi, Manoochehr Makvandi, Azadeh Haghi Navand, Shokouh Shayanpour, Heshmatolah Shahbazian, Kimia Makvandi, Saeed Eynali Varnosfaderani, Sepideh Nasimzadeh Pages 759-764
    Background and Objectives

    Hepatitis E Virus (HEV) account for acute hepatitis, fulminant liver failure and chronic hepatitis worldwide. Several high risk groups including hemodialysis (HD) patients are at risk of HEV infection. Based on consequences of HEV infection it is important to determine the serological and molecular epidemiology of HEV in HD patients. The aim of this study was to evaluate the frequency of HEV antibodies and HEV RNA in HD patients.

    Materials and Methods

    The sera of 84 HD patients were collected and tested for anti- HEV IgG and anti IgM antibodies using enzyme-linked immunosorbent assay (ELISA) at Golestan hospital in Ahvaz city during October 2014 and November 2014. HEV RNA was tested in HD patients using RT PCR. The prevalence of anti- HEV IgG was evaluated in the age group (52/84) > 50 and (31/84) < 50 years.

    Results

    Out of 84 patients, 52 (61.9%) were males and 32 (38.1%) females. The mean age of participants was 52 ± 1.57 years. 43/84 (51.19%) cases including 26/52 (50%) males and 17/32 (53.1%) females were positive for anti-HEV IgG (p=0.95). Among the 43 cases positive anti-HEV IgG 8 cases including 5 (9.61%) males and 3 (9.37%) females tested positive for anti-HEV IgM (p=0.729) while the HEV RNA was negative in HD patients. The distribution of anti- HEV IgG was 62.75% and 33.33% among the age group >50 and <50 respectively (p=0.015).

    Conclusion

    This study showed high prevalence of anti-HEV IgG antibodies (51.19%) were observed among the HD patients while the HEV RNA tested negative in HD patients. The rate of HEV IgG is significantly higher with increased age. Further investigation require to identify the factors account for high seroprevalence of HEV in Ahvaz HD units.

    Keywords: Hepatitis E virus, Hemodialysis patients, Enzyme-linked immunosorbent assay
  • Maryam Sarkardeh, Ali Ahmadabadi, Zahra Sadrzadeh, Javad Koushki, Amir Esmaeili, Sima Davoodi, Mostafa Izanlu Pages 765-769

    Necrotizing fasciitis and myonecrosis caused by Escherichia coli is an extremely uncommon infection with a high mortality. We present a case of 41-year-old man with no history of underlying disease one week after covid-19 infection, who was admitted with symptoms of Fournier's gangrene and then E. coli-induced monomicrobial necrotizing fasciitis.

    Keywords: Necrotizing fasciitis, COVID-19, Escherichia coli, Vacuum-assisted closure