Jundishapur Journal of Microbiology

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In Collaboration with Ahvaz Jundishapur University of Medical Sciences

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Jundishapur Journal of Microbiology, (JJM) is dedicated to the publication of manuscripts on topics concerning all aspects of microbiology. The topics include medical, veterinary and environmental microbiology, molecular investigations, and infectious diseases. Aspects of immunology and epidemiology of infectious diseases are also considered.

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Helicobacter Pylori Infection and Non-Steroidal Anti-Inflammatory Drugs Mediated Gastric Ulcer: Insights into Efficacy and Strategies for Gastroprotective Treatments and Disease Management.
(Brieflands, 2024-11-30) Zhitiao Lv; Shaoyu Fang; Yan Wang
Background: Helicobacter pylori infection and the use of non-steroidal anti-inflammatory drugs (NSAIDs) are the primary etiological factors contributing to gastric ulcer disease, characterized by mucosal erosions in the stomach. This condition remains a significant global health challenge, particularly in low- and middle-income countries, where it substantially contributes to morbidity and premature mortality. Complications such as gastrointestinal bleeding and perforation exacerbate the disease burden, underscoring the need for effective preventive and therapeutic strategies.
Study of bacteria isolated from urinary tract infections and determination of their susceptibility to antibiotics
(Brieflands, 2009-07-31) Mansour Amin; Manijeh Mehdinejad; Zohreh Pourdangchi
Introduction and objective: Approximately 1 in 3 women will require antimicrobial treatment for a Urinary Tract Infection (UTI) before age 24, and 40% to 50% of women will have a UTI during their lifetime. UTIs in male patients are considered complicated. Escherichia coli is the most common cause of UTIs. Materials and methods: In the present study 7056 patients with clinical symptoms and suspected to UTI were sampled. Clean-Catch midstream urine of the patients was collected. Urine specimens were cultured for isolation of the microbial agents of UTI. The isolated bacteria were identified using biochemical tests. Disk diffusion susceptibility test was used to determine susceptibility of bacterial agents to antibiotics. Results: In this study 553(8.7%) patients out of 7056 were shown to be urine culture positive (68% females and 32% males). The most isolated bacterium was E. coli with frequency rate of 59%. The other bacteria were Klebsiella spp. (11.6%), Enterobacter spp. (9.8%), Pseudomonas spp. (7.2 %), Proteus spp. (2.9%), Acinetobacter spp. (2.7%), Congolese positive Staphylococci (2.2%), Coagolase negative Staphylococci (2.3%), Citrobacter spp. (1.3%) and Streptococci α hemolytic (1.1%). All Gram-negative bacteria were more sensitive to amikacin (90.5-100%). The Gram-positive cocci isolated were more sensitive to tobramycin, kanamycin and ciprofloxacin (100%). Conclusion: It is concluded that Gram-negative bacilli were responsible for UTI infections in our patients. The most common isolated bacteria from urinary tract infections were E. coli and the most effective antimicrobial agents were amikacin, tobramicin and ciprofleoxacin against Gram-negative bacilli and also the most effective antibiotics against Gram-positive cocci were kanamicin, tobramicin and ciprofleoxacin.
Antibiotic Resistance Pattern Among Gram Negative Mercury Resistant Bacteria Isolated From Contaminated Environments
(Brieflands, 2013-12-01) Nima Mirzaei; Hilda Rastegari; Mehdi Kargar
Background: Mercury is one of the most toxic heavy metals. Even a small amount of it is toxic for all living organisms. Some of bacteria have developed special resistance mechanisms against mercury, in addition to resistant to different antibiotics. These bacteria usually acquire Hg and antibiotic resistance genes via horizontal gene transfer in their habitat. Objectives: The aim of this study was isolation and identification of mercury resistant bacteria and evaluating the relation between increase of environmental levels of mercury and prevalence of antibiotic resistance among Gram negative Hg resistant bacteria. Materials and Methods: The samples were collected from water and sediments of Kor River. We evaluated amounts of mercury in the water and sediment samples and counted the number of bacteria in both Hg containing and non-Hg containing media. Antibiotic resistance pattern was studied using disk diffusion method for Hg resistant and Hg sensitive bacteria. Results: The frequencies of mercury resistant bacteria were between 35% for Pole Khan station and 2.5% for Doroodzan station. These stations were the most and the lowest mercury-contaminated areas of the Kor River respectively. Pseudomonas sp., Entrobacter sp., Escherichia coli, Klebsiella sp. And Serratia marcescens were isolated as mercury resistant bacteria. The highest level of antibiotic resistance was seen for ampicillin, tetracycline and sterptomycin. Antibiotic resistance frequencies among these bacteria were higher than mercury sensitive ones. Conclusions: Our results showed that Hg resistance genes in contaminated areas are exchanged between residing bacteria along with the antibiotic resistance genes and resulted in prevalence of antibiotic resistance among residing bacteria.
Antibiotic Resistance Pattern Among Gram Negative Mercury Resistant Bacteria Isolated From Contaminated Environments
(Brieflands, 2013-12-01) Nima Mirzaei; Hilda Rastegari; Mehdi Kargar
Background: Mercury is one of the most toxic heavy metals. Even a small amount of it is toxic for all living organisms. Some of bacteria have developed special resistance mechanisms against mercury, in addition to resistant to different antibiotics. These bacteria usually acquire Hg and antibiotic resistance genes via horizontal gene transfer in their habitat. Objectives: The aim of this study was isolation and identification of mercury resistant bacteria and evaluating the relation between increase of environmental levels of mercury and prevalence of antibiotic resistance among Gram negative Hg resistant bacteria. Materials and Methods: The samples were collected from water and sediments of Kor River. We evaluated amounts of mercury in the water and sediment samples and counted the number of bacteria in both Hg containing and non-Hg containing media. Antibiotic resistance pattern was studied using disk diffusion method for Hg resistant and Hg sensitive bacteria. Results: The frequencies of mercury resistant bacteria were between 35% for Pole Khan station and 2.5% for Doroodzan station. These stations were the most and the lowest mercury-contaminated areas of the Kor River respectively. Pseudomonas sp., Entrobacter sp., Escherichia coli, Klebsiella sp. And Serratia marcescens were isolated as mercury resistant bacteria. The highest level of antibiotic resistance was seen for ampicillin, tetracycline and sterptomycin. Antibiotic resistance frequencies among these bacteria were higher than mercury sensitive ones. Conclusions: Our results showed that Hg resistance genes in contaminated areas are exchanged between residing bacteria along with the antibiotic resistance genes and resulted in prevalence of antibiotic resistance among residing bacteria.
Effects of Onion Juice on the Normal Flora of Eyelids and Conjunctiva in an Animal Model
(Brieflands, 2014-05-01) Mahmoud Nejabat; Alireza Salehi; Parisa Noorani Azad; Mohammad Javad Ashraf
Background:: Traditional medicine/complementary alternative medicine may suggest new ideas to modern medicine in order to face new challenges however these concepts should be acknowledged based on experimental studies. Objectives:: We aimed to study the effects of onion (Allium cepa) juice on the normal flora of conjunctiva and eye lids, and to follow the histopathology changes of conjunctiva in an animal study. Materials and Methods:: Twenty-four rabbits were randomly classified into three equal groups. Groups 1, 2 and 3 received fresh red onion juice, as an eye drop, into the right eye twice daily for; one week, one month, and two months, respectively. Microbiological sampling by sterile swabs was performed before and after the intervention. Cultural characteristics, including the growth rate and the kind of organisms, are reported. At the end of the study, pathological samples were collected from the inferior fornix. Results:: After the intervention, the number of positive cultures in the samples, collected from both the conjunctiva and eyelid, had decreased. Group 3 demonstrated the lowest amount of growth after the administration of the onion juice and the bacterial isolation rates from each organism had decreased. All pathological samples revealed some degree of inflammation. There was no evidence of metaplasia or dysplasia. There was no significant difference between the growth rates of organisms in the experimental groups using statistical analysis. Conclusions:: According to our experiment, onion has an inhibitory effect on the growth of normal eye flora; although the duration of onion juice instillation did not show any significant effect on the group results. Hence, this finding is an initiating point for further investigations into the antimicrobial properties of this herb to treat common eye infections, including conjunctivitis and blepharitis.
Molecular Detection of Class-D OXA Carbapenemase Genes in Biofilm and Non-Biofilm Forming Clinical Isolates of Acinetobacter baumannii
(Brieflands, 2015-01-01) Omid Azizi; Mohammad Reza Shakibaie; Farzan Modarresi; Fereshteh Shahcheraghi
Background: Emergence and spread of carbapenemase (blaOXA) genes in multidrug resistant Acinetobacter baumannii (MDR-AB) forming biofilm complicated treatment of the patients infected with this microorganism particularly in intensive care units (ICUs). Objectives: The current study aimed to determine the prevalence of molecular class-D OXA carbapenemase in biofilm and non-biofilm forming strains of MDR-AB. Materials and Methods: A total of 65 strains of MDR-AB were isolated from the patients hospitalized in the ICU of two hospitals in Kerman, Iran. The isolates were identified by conventional microbiological tests as well as API 20NE assay. Antibiotic susceptibility was carried out by disk diffusion method; minimum inhibitory concentration (MIC) of carbapenems was measured by E-test. The presence of blaOXA genes among the isolates were studied by duplex-polymerase chain reaction and application of appropriate primers. Biofilm formation was detected by microtiter plate method. Results: The isolates were highly resistant to ciprofloxacin, levofloxacin, piperacillin, nalidixic acid and third generation cephalosporins such as tigecycline (7%; n = 5) and colistin (13%; n = 8). Among the isolates, 77% (n = 50) exhibited high MIC (265μg/mL) for imipenem. Both the blaOXA-51 and blaOXA-23 like genes coexisted in all the isolates; while, blaOXA-24/40 like gene was only detected in 29 imipenem-resistant strains (P ≤ 0.05). The blaOXA-58 like gene was not detected among the isolated strains. Quantification of biofilm introduced 23 isolates (including blaOXA-24/40 strains) with efficient attachment to microtiter plate; while, those isolates without blaOXA-24/40, or imipenem-sensitive strains formed weak or no biofilm. Conclusions: Coexistence of the blaOXA-51, blaOXA-23 and blaOXA-24/40 like genes, along with formation of strong biofilm, in MDR-AB strains particularly with indiscriminate use of imipenem, complicated treatment of the patients infected with these bacteria in the hospitals understudy.
Necrotic Response to Low Pathogenic H9N2 Influenza Virus in Chicken Hepatoma Cells
(Brieflands, 2015-01-01) Seyedeh Zahra Mosavi; Shahla Shahsavandi; Mohammad Majid Ebrahimi; Ali Reza Hatami; Kaveh Sadeghi; Hassan Shahivandi
Background: Limited knowledge about the molecular mechanism of avian influenza H9N2 virus pathogenicity in birds as well as human hosts has limited the development of effective control against the disease. To overcome this issue detailed understanding of the infectious characteristics of the virus in host cells should be obtained. Objectives: In this study we examined the replication kinetics of H9N2 virus in a chicken hepatoma cell line to obtain insight into the pathogenesis of H9N2 viruses. Materials and Methods: The kinetic replication of H9N2 influenza virus in chicken hepatoma and fibroblastic cells was studied in the presence and absence of supplemental trypsin. High viral titers observed in liver cells in a short time correlated with the degree of cytopathic effects. To determine whether the ultimate outcome of infection results in programmed cell death, the infected cells were observed by the cell viability assay, DNA fragmentation, caspase cascade activation, and quantified lactate dehydrogenase release. Results: The degree of viability was significantly reduced in infected hepatoma cells. Observations of caspase activation and cell DNA laddering in infected cells were not indicative of apoptosis. The infected hepatoma cells released lactate dehydrogenase, which is consistent with cell death by necrosis. Conclusions: Taken together, these data reveal that cellular protease of chicken liver cells allows the replication of high yields of H9N2 virus in the absence of trypsin and also cell death in the infected cells is due to necrosis.
Construction and Immunogenicity Analysis of Hepatitis C Virus (HCV) Truncated Non-Structural Protein 3 (NS3) Plasmid Vaccine
(Brieflands, 2016-03-01) Mohammad-Hassan Pouriayevali; Taravat Bamdad; Mohammad-Reza Aghasadeghi; Seyed Mehdi Sadat; Farzaneh Sabahi
Background: To develop hepatitis C virus (HCV) vaccine, induction of potent humoral and T cell response against immunogenic targets with conserved region should be achieved. T cell response against NS3 is often associated with complete clearance of the virus. Objectives: Herein, we expressed the truncated form of NS3 in a mammalian cell line and evaluated immune responses of NS3 DNA vaccine in BALB/c. Materials and Methods: The partial length of NS3 gene, which encodes immunogenic epitopes (1095 - 1379 aa), was amplified by reverse transcription-polymerase chain reaction (RT-PCR) on RNA obtained from a patient with HCV, inserted into pcDNA3.1 plasmid using XhoI/HindIII sites, and finally evaluated by restriction analysis and sequencing. After transfection of the recombinant plasmid into HEK293T cells, the NS3 protein expression was confirmed by western blotting. Mice were immunized intra-dermally close to the base of the mice tail with four doses in two-weeks intervals and the immune responses were assessed using total and subtypes of IgG antibody assay, cell proliferation and cytokine assay. Results: The pcDNA3.1 plasmid harboring the coding sequence of NS3 (pc-NS3) was constructed and confirmed with the expected size. Proper expression of the recombinant protein in transfected HEK 293T cells was confirmed using western blotting. The immunization results indicated that pc-NS3 induced significant levels of total antibody, IgG2a subclass antibody, Interferon (IFN)-γ, Interleukin (IL)-4 and proliferation assay compared to the control group (P < 0.05). Conclusions: The pc-NS3 possesses the capacity to express NS3 in the mammalian cell line and demonstrated strong immunogenicity in a murine model. Our primary results demonstrated that the immunogenic truncated region of NS3 could be used as a potential vaccine candidate against hepatitis C.
Molecular Characterization of Carbapenem-Resistant Klebsiella pneumoniae Species Isolated From a Tertiary Hospital, Ankara, Turkey
(Brieflands, 2017-10-31) Nevreste Celikbilek; Ozlem Unaldi; Fisun Kirca; Aysegul Gozalan; Ziya C Acikgoz; Riza Durmaz
Background: Carbapenem-resistant Klebsiella pneumoniae isolates can spread among the hospitalized patients and result in serious infections and increase mortality. Molecular studies provide useful information about resistance mechanisms and cross-transmission among the resistant isolates. Objectives: The current study aimed at investigating phenotypic and molecular characteristics of the carbapenem resistance and clonal relationship among the K. pneumoniae isolates recovered from infection sites and rectal swabs of the patients hospitalized in a tertiary hospital. Methods: Resistance to carbapenems was investigated by disc-diffusion and E-test methods. Modified Hodge test (MHT) and ethylenediaminetetraacetic acid (EDTA)-combine disc (ECD) methods were performed to determine carbapenemases. Carbapenemase-encoding genes including blaOXA-48, blaNDM-1, blaKPC, blaIMP, and blaVIM were investigated by multiplex polymerase chain reaction (PCR). Clonal relationship among the isolates was determined by the pulsed-field gel electrophoresis (PFGE). Results: Carbapenem resistance was observed in 93 (5.8%) of the 1605 K. pneumoniae isolates. Majority (n = 66, 71%) of the resistant isolates were recovered from the patients in intensive care units (ICUs). Almost all carbapenem-resistant K. pneumoniae isolates (n = 91, 97.8%) were positive with MHT. Only 6 (6.5%) of the resistant isolates were positive with ECD method. The blaOXA-48 and blaNDM-1 were found in 90.3% (n = 84) and 6.5% (n = 6) of the resistant isolates, respectively. The pulsed-field gel electrophoresis yielded 55 different pulsotypes. Fifteen pulsotypes were identified as clusters with ≥ 2 isolates and a clustering rate of 57%. Based on the similarity coefficient higher than 85%, a total of 75 (80.6%) isolates were clonally related to each other. Conclusions: The high rate of carbapenem resistance and 80.6% clonal relationship between the isolates collected in a 3-year period indicated a serious potential threat for the hospital-acquired infections and pointed longtime cross-contamination.
Risk Factor Analysis of Brucellosis in Hulunbuir, China, Using the Propensity Score Matching Method
(Brieflands, 2021-06-30) Li Peng; Xiuwen Liang; Lei Zhu; Chen Liang; Chenfang Liu; Xiaohui Chi
Background: Brucellosis is the most widespread zoonosis worldwide and one of the most neglected zoonotic diseases. At present, large-scale farms are growing rapidly, increasing the risk of disease transmission. Objectives: In this study, the propensity score matching (PSM) method was used to analyze the epidemiological characteristics of brucellosis and explore the risk factors of brucellosis infection in Hulunbuir, Inner Mongolia, China. Methods: A questionnaire for brucellosis was designed based on general knowledge and the protection of key groups of brucellosis. Epidata 13.0 software was used to establish the questionnaire, and propensity score matching was used to select cases that met the requirements of case-controls. Results: A total of 152 cases and 456 controls were included. The results of the study show that feeding livestock, carrying lambs regularly, and raising livestock without protective measures can increase the risk of brucellosis infection. Conclusions: Behavioral factors are the main risk factors for brucellosis, and livestock keepers should strengthen self-protection when working.
Levels of Transforming Growth Factor-Beta After Immunization of Mice With in vivo prepared Toxoplasma gondii Excretory/Secretory Proteins
(Brieflands, 2015-05-31) Seyed Hossein Abdollahi; Fateme Ayoobi; Hossein Khorramdelazad; Behzad Nasiri Ahmadabadi; Mohammadtaghi Rezayati; Mohammad Kazemi Arababadi; Mohammad Zare-Bidaki
Background:: Zoonotic parasite Toxoplasma gondii has a high prevalence in human populations. A suitable vaccine for animals can stop the transmission of infection between animal and human. Objectives:: The aim of this study was to evaluate in vivo prepared excretory/secretory antigens (E/SA) as a potential candidate for immunization against the parasite and its effect on the production of transforming growth factor-beta (TGF-β). Materials and Methods:: Toxoplasma gondii tachyzoites were inoculated in the peritoneal cavity of mice and E/SA was harvested and used in animal immunization with and without adjuvant. Serum levels of anti-E/SA antibodies and TGF-β were measured in days 0, 3, 7, 14, 28 and 56 after immunization using ELISA technique. The measurements were statistically analyzed. Results:: Our results showed that the serum levels of anti-E/SA immunoglobulins significantly increased in all of the immunized groups. The differences of the serum levels of TGF-β between the groups were statistically significant at days 28 and 56 after immunization with E/SA. Conclusions:: Based on our study, in vivo prepared E/SA may be considered as a good candidate for animal immunization.
A Bibliometric Analysis of Microbiology Publications in Sub - Saharan Africa during Years 2000 to 2014
(Brieflands, 2018-03-31) Yashik Singh
Context: Research, development, and innovation are becoming increasingly important in the rapidly changing political, financial, and social landscape of the world. It is important to understand the landscape of research and development in developing courtiers, as research and development has been shown to improve developing countries’ economic growth. The aim of this study was to obtain a snap shot of a bibliometric analysis of the publications in the field of microbiology in sub - Saharan Africa (SSA). Among other questions, this study aimed at determining the most published authors, the most common areas of research, and institutes with the greatest number of publications in this field. Evidence Acquisition: This study was conceptualised using 3 steps: creating a search strategy that encompasses the major fields of microbiology, creating and executing a search query, and analysing the results. Scopus was chosen as the search engine since it is the largest database of peer - reviewed literature and includes original articles, reviews, conferences, letters, editorials, and articles - in - press, etc. Conclusions: Although at first glance it may seem that SSA contributed very little towards (less than 2%) the worlds microbiology literature, a closer examination of gross domestic product (GDP) spent on research proves that SSA countries are making inroads in publishing literature. Most literature published over the last 14 years has been journal articles in journals with an impact factor of 3.7. In 2014, most articles were published in journals with an average impact factor of 6.1. In terms of the number of publications by the top authors in the field, it seems as if they quantitatively match other international countries like Brazil and India.
Characterization of Proteus mirabilis Isolated from Patient Wounds at Bolan Medical Complex Hospital, Quetta
(Brieflands, 2019-07-23) Umbreen Zafar; Muhammad Kamran Taj; Imran Nawaz; Asma Zafar; Imran Taj
Background: Wound infection is the cause of mortality and morbidity on a global scale. Microorganisms infecting wounds can multiply and colonize in the wound, resulting in host tissue damage. Objectives: The present study was conducted to identify Proteus mirabilis in wounds of patients in Quetta district. Methods: This study was conducted from June 2017 to June 2018 at the Center for Advanced Studies in Vaccinology and Biotechnology, University of Balochistan, Quetta. Overall, 480 different wound samples were collected from patients admitted to Bolan Medical Complex Hospital, Quetta. Proteus mirabilis was isolated using differential and selective media and characterized by biochemical tests (catalase, oxidase, IMViC, and sugar fermentation), antimicrobial susceptibility tests, and PCR. Results: There were 64 (13.3%) samples positive and 416 (86.6%) samples negative for P. mirabilis. The results showed that wounds infected with P. mirabilis were more common in male patients (n = 40; 8.3%) than in female patients (n = 24, 5%). The age distribution showed that the infection of wounds with P. mirabilis was the highest in 16 - 30-year-old group (n = 32; 6.70%), followed by the age groups of 5 - 15 (n = 24; 5%) and 30 - 50 years (n = 8; 1.60%). Diabetic (n = 24; 5%) and surgical (n = 24; 5%) wounds were more affected by P. mirabilis than burn wounds (n = 16; 3.30%). Proteus mirabilis was sensitive to gentamicin (n = 50; 78%) and amikacin (n = 53; 82.8%) but resistant to penicillin G (n = 58; 90%), ampicillin (n = 56; 87.5%), amoxicillin (n = 60; 93.7%), cefuroxime (n = 61; 95.3%), ceftriaxone (n = 57; 89%), ceftazidime (n = 59; 92.1%), imipenem (n = 62; 96.8%), ciprofloxacin (n = 55; 85.9%), and tetracycline (n = 59; 92%). The PCR-based identification of P. mirabilis showed clear bands of 533 bp of the ureC1 gene. Conclusions: The pathogenesis of P. mirabilis in wound infection and its antimicrobial sensitivity are major problems worldwide. The use of aminoglycosides such as gentamycin and amikacin is effective against P. mirabilis and can help prevent the spread of infection and reduce the cost of treatment. The PCR technique is one of the sensitive, timesaving, specific, and cost-effective ways for the identification of the pathogenic genes of P. mirabilis.
Diagnostic value of hydatid cyst antigens using western blotting method
(Brieflands, 2010-10-31) Behzad Haghpanah; Badrossadat Mosavat; Zahra Ghayour; Farzad Oreizi
Introduction and objective: Hydatidosis is one of the most important and commonly found parasitic zoonoses in both humans and different animals, which is caused by the cestode helminthes Echinococcus granulosus. The diagnosis of the disease is primarily based on imagery techniques. Thus, highly sensitive and reliable serologic methods are required to confirm the diagnosis. AntigenB (AgB) and protoscoleces antigen (PSC Ag) were purified as two specific parasitic antigens and then evaluated against sera from two groups of hydatidosis and non-hydatidosis (control) subjects using the western blotting method in order to identify the most sensitive and specific antigen. Materials and methods: Sera samples were taken from 22 patients under operation for hydatid cyst. 16 patients were also included as control group. Cyst fluid and protoscoleces were extracted and partially purified in a protein A column. Using SDS-PAGE, subunits of the cyst fluid antigen, AgB, and PSC Ag were identified. Finally, the subunits were transferred from gel to nitrocellulose membrane in a western blot test and reacted with hydatid and control sera in order to assess their sensitivity and specifity. Results: Three antigens were identified as the subunits of AgB while 10 antigens were identified as PSC Ag. The sensitivity and specifity of AgB subunits in the western blot test were 77% and 100%, respectively. None of the PSC Ag subunits had both high sensitivity and high specifity concurrently. Conclusion: It has been shown by the western blot test that the AgB 8/12 and 16 KDa subunit components had high diagnostic sensitivity and specifity levels (81% and 100%, respectively) and that they could presumably assist the physician in his pre- and post-operation diagnosis of hydatid cysts.
Prokaryotic Expression and Monoclonal Antibody Preparation of Rabies Virus Phosphoprotein
(Brieflands, 2017-08-31) Juan Hu; Xueshan Xia; Ming Yang; Yuzhu Song; Qinqin Han; Qiang Chen; Jie Zan; Jinyang Zhang
Background: Rabies is a zoonotic infectious disease that infects the human and animal central nervous system. Worldwide, especially in low-income countries, this disease is still a burden for public health. Among the rabies virus proteins, phosphoprotein plays a very important role in viral infection, and this research found that immunization of rabies virus vaccines could widely induce antibody responses against phosphoprotein, therefore rabies virus phosphoprotein may be a useful target for development of rapid and low cost serological diagnosis test, and therapeutic drugs. Objectives: The aim of this study was to prepare anti-rabies virus phosphoprotein monoclonal antibody, which is used for rapid detection and diagnosis of rabies virus infection. Methods: The phosphoprotein gene was amplified by the polymerase chain reaction (PCR), and sub-cloned in a his-tagged prokaryotic expression vector to generate a recombinant plasmid named pET-32a-RABV-P. Recombinant RABV-P was induced by isopropyl-β-D-Thiogalactopyranoside (IPTG), and then purified by the Ni-NTA purification system. Immunization of BALB/c mice with the recombinant protein was performed, and the spleen cells of the immunized mice and SP2/0 myeloma cells were fused together to obtain the monoclonal cell strains, and then identification of the characteristics of the antibody by the enzyme linked immunosorbent assay (ELISA), western blotting, and indirect immunofluorescence assay was done. Results: The prokaryotic expression vector of pET-32a-RABV-P was successfully constructed. The fusion protein was expressed in Escherichia coli Rosetta and purified. After cell fusion, a hybridoma cell line 1A4 was successfully obtained. The antibody titer of the anti-RABV-P ascites reached 256,000. The results of western blotting and indirect immunofluorescence assay indicated that 1A4 hybridoma cell line was able to produce specific antibodies against rabies virus phosphoprotein. Conclusions: Recombinant rabies virus phosphoprotein could be successfully expressed in E. coli. A specific monoclonal antibody against rabies virus phosphoprotein has been successfully prepared.
Identification of a Bacterium Isolated from Soil of Ahvaz Contaminated by Oil and Determination of its Protease Stability in Organic Solvents
(Brieflands, 2013-02-28) Gholamhossein Ebrahimipour; Hossein Sadeghi; Zahra Khosravi babadi; Soleiman Amiri
Background: Proteases are often used as catalysts for peptide synthesis. To shift the thermodynamic equilibrium in favor of the peptide synthesis, reaction media should contain organic solvent. However, known enzymes are usually inactivated by adding organic solvents to reaction media. Objectives: In this study, we reported a bacterium isolated from soil of Ahvaz, Iran contaminated by crude oil producing an organic solvent-stable protease, and the effect of organic solvents on proteolytic activity was investigated. Materials and Methods: Isolated bacterium was cultured in mineral salt medium containing glucose and peptone. After 48 hours incubation at 35 C and 130 rpm, culture media were centrifuged and resulted supernatant filtered using 0.22 ?m nitrocellulose membrane filter. Proteolytic activity of supernatant was determined by Keay and Wildi method (1970) by using casein as substrate. The effects of different concentrations of various organic solvents including acetone, ethanol, pentanol, cyclohexane, benzene, n-hexane, and n-decane and, also, the effects of temperature and pH on protease stability and activity were examined. Results: According to 16SrDNA sequencing, strain ISA9 was identified as a new strain of Bacillus licheniformis. This strain was able to produce an extracellular organic solvent- tolerant protease. After 30 minutes incubation at 37 C, caseinolytic activity of crude protease was increased in 25 and 50% of acetone, ethanol, benzene, cyclohexane, and hexane compared to non-solvent control. The enzyme was also activated 1.64, 1.23, and 1.17 times by 75% (v/v) of benzene, decane, and hexane, respectively. The protease was active in a broad range of pH (from 6 to 10) with the optimum pH 10. The optimum temperature for the activity of this protease was 70 C and the enzyme remained active after incubation at 30-50 C for 30 minutes. Conclusions: In this study, we isolated B. licheniformis producing an organic solvent-stable protease from oil-contaminated soil. The protease was stable and active in various organic solvents. By purification, the protease could be used as a biocatalyst for organic solvent-based enzymatic synthesis.
Bioassay Guided Fractionation of an Anti-Methicillin-Resistant Staphylococcus aureus Flavonoid From Bromus inermis Leyss Inflorescences
(Brieflands, 2014-12-01) Atousa Aliahmadi; Fateme Mirzajani; Alireza Ghassempour; Ali Sonboli
Background: Plants are considered as promising sources of new antibacterial agents as well as bioassay guided fractionation. Objectives: In the present work, the antibacterial properties, especially against methicillin-resistant Staphylococcus aureus (MRSA), of Bromus inermis inflorescence was studied, using the bioassay guided fractionation as well as the bio-autographic method. Materials and Methods: The plant organic extract was prepared via maceration in methanol, followed by the fractionation using n-hexane. The extracts were subjected for minimum inhibitory concentrations (MICs) against some human pathogenic bacteria via standard broth micro-dilution assay. Thereafter, a bio-autographical method was applied using the high performance thin layer chromatography (HPTLC) coupled with agar overlay assays for the primary characterization and identification of bioactive substance (s). Results: Through the bioassay guided fractionation method, the greatest antibacterial activities were related to the n-hexane extract. It was also revealed that the effective anti-MRSA agent of the assessed plant was a relatively polar substance with an MIC value of about 8 μg/mL against the tested MRSA strain (in comparison with the MIC value of 32 μg/mL for chloramphenicol). Conclusions: As a result of the full range UV-Vis scanning of the responsible band in the HPTLC experiments (200-700 nm), the flavonoid was the most imaginable natural compound.
Multi-Drug Resistant Acinetobacter-Derived Cephalosporinase and OXAsetC Genes in Clinical Specimens of Acinetobacter spp. Isolated From Teaching Hospital
(Brieflands, 2013-03-02) Reza Khaltabadi Farahani; Rezvan Moniri; Kamran Dastehgoli Farahani
Background: Hospital-acquired infections caused by multi-drug resistant Acinetobacter spp. are often extremely difficult to treatand this has proved to be a serious problem worldwide. Objectives: The aim of this study was to determine the incidence rates and distribution patterns of multi-drug resistant (MDR) Acinetobacter spp. strains and the occurrence of Acinetobacter-derived cephalosporinase (ADC-7) and OXA-type carbapenemases (OXAsetC genes) in clinical specimens in the Beheshti Teaching Hospital in Kashan, Iran. Materials and Methods: This descriptive study was carried out on sixty isolates of Acinetobacter spp. and clinical samples collected from patients. The level of antibiotic resistance was determined by the disc diffusion method and the results were interpreted according to the Clinical and Laboratory Standards Institute (CLSI) procedure. Polymerase chain reaction (PCR) amplification of the genetic determinants of resistance was also determined. Results: The resistance rates were; amikacin (80%), tobramycin (68.3%), ceftazidime (60%), ciprofloxacin (55%), piperacillin/tazobactam (51.7%), doxycycline (50%), SXT/TMP (sulfamethoxazole / trimethoprim)(48.3%), levofloxacin (43.3%), gentamicin (40%), imipenem (25%), and sulbactam/ampicillin (20%). The frequency of MDR Acinetobacter spp. strains isolated was found to be 56.7%. These isolates were most sensitive to imipenem followed by ampicillin/sulbactam and gentamicin. The prevalence of genes for ADC-7 and OXAsetC in the Acinetobacter spp. was; 34 (56.7%) and 32 (53.3%), respectively. The positive percentages of MDR isolates for ADC-7 were 82.4% and for OXAsetC they were 73.5%. Conclusions: Our phenotypic analysis demonstrated that Acinetobacter spp. isolates were resistant to most clinically significant antibiotic classes. This is the first report concerning Acinetobacter-derived cephalosporinase, blaADC, enzymes in Acinetobacter spp. isolates from Iran.
Associations Between Fusobacterium nucleatum and msh2, mlh1, and msh6 Gene Expression in Colorectal Cancer
(Brieflands, 2024-02-29) Reza Torkashvand; Bahareh Hajikhani; Reza Hosseini Doust; Hossein Dabiri; Masoud Dadashi
Background: Colorectal cancer (CRC) is a major global health concern, and the link with Fusobacterium nucleatum has received considerable attention. Objectives: This study aimed to explore the prevalence of F. nucleatum and to assess the expression of the msh2, mlh1, and msh6 genes in CRC patients compared to a control group using real-time PCR. Methods: Forty CRC patients and twenty individuals from a control group participated in this study. Gastroenterologists collected biopsy specimens from which DNA and RNA were extracted using a specialized tissue extraction kit. Complementary DNA (cDNA) was then synthesized. Real-time PCR was employed to evaluate the expression levels of the msh2, mlh1, and msh6 genes and the presence of the F. nucleatum-specific 16srRNA gene to determine the relative prevalence of this bacterium in each group. Results: Results indicated a higher prevalence of the F. nucleatum-specific 16srRNA gene in CRC patients than in the control group. Additionally, expression levels of the msh2, mlh1, and msh6 genes were significantly higher in the cancer group, suggesting their role in CRC pathogenesis. The distribution of F. nucleatum was particularly high in the sigmoid and rectum areas of the colon. Conclusions: This study underscores the significance of F. nucleatum in CRC and provides insights into its association with altered gene expression patterns. Understanding the prevalence of F. nucleatum and its impact on msh2, mlh1, and msh6 genes may aid in developing improved diagnostic and therapeutic strategies for CRC. Further research is necessary to elucidate these relationships more comprehensively.
Role of Oxidative Stress Response and Trehalose Accumulation in the Longevity of Fission Yeast
(Brieflands, 2015-06-01) Bedia Palabiyik; Farinaz Jafari Ghods
Background: Glucose is the preferred carbon and energy source in most organisms and plays an active role in the regulation of many biological processes. However, an excess of glucose leads to such undesirable conditions as diabetes and age-related diseases. Since Schizosaccharomyces pombe homologous of many human genes, it offers several advantages for the investigation of the molecular mechanisms underlying human disease and aging studies. We have identified two glucose-repression-resistant mutants (ird5 and ird11) of S. pombe. Objectives: We aimed to investigate the possible relationship between lifespan extension and oxidative stress response induced by exposure to hydrogen peroxide alongside the trehalose accumulation level by using the two S. pombe mutants (i.e. ird5 and ird11), which are repressed by glucose and are resistant to oxidative stress. Materials and Methods: We employed trehalose accumulation measurement and colony-forming unit (CFU) counting using the ird mutants in exponential and stationary phases and compared them to the wild type grown in repressed, de-repressed, and stressed conditions to clarify the possible relationship between glucose signaling, oxidative stress response, and lifespan in S. pombe. Results: The lifespan of the ird5 mutant was significantly longer that of either the ird11 mutant or the wild type cells. Under repressed condition, the trehalose content was increased remarkably on the 3rd day of the study in the ird11 mutant and the wild type. Under de-repressed condition, the level of intracellular trehalose was notably increased on the 3rd day in ird11. Under stressed condition, the trehalose level in ird11 was increased on the 3rd day as a pattern similar to that observed in the wild type. Conclusions: Our results demonstrated no significant correlation between the ird5 lifespan and the trehalose concentration. Likewise, the correlation between lifespan extension, trehalose accumulation, and cellular resistance to hydrogen peroxide was not significant.

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