Medical Laboratory Journal

Prevalence of Plasmid-Mediated Quinolones Resistance among Klebsiella pneumoniae Strains Isolated from Hospitals in Borujerd, Iran

Leili Shokoohizadeh

ABSTRACT
       Background and Objective: Klebsiella pneumoniae is one of the most common causes of bacterial infections. Presence of plasmid-mediated quinolone resistance genes causes low level of resistance in K. pneumoniae. This study investigated the prevalence of resistance to quinolones and fluoroquinolones, and the frequency of qnrA, qnrB and qnrS genes among K. pneumoniae strains.
        Methods: The study was performed on 100 K. pneumoniae strains isolated from hospitals in city of Borujerd (Iran) during April to September 2014. Susceptibility of the isolates to nalidixic acid, ciprofloxacin, norfloxacin and ofloxacin was evaluated. Minimum inhibitory concentration (MIC) of ciprofloxacin was determined using ciprofloxacin Etest strips. Polymerase chain reaction was performed to detect qnrA, qnrB and qnrS genes in quinolone-resistant isolates using specific primers.
      Results: The results showed that 38% of the isolates were resistance to both nalidixic acid and ciprofloxacin. The prevalence of ofloxacin- and norfloxacin-resistant isolates was determined to be 18% and 15%, respectively. The MIC values for ciprofloxacin were ranging from 0.064 to ≥256 μg/ml. In addition, four ciprofloxacin-resistant isolates (10%) had MIC of ≥256 μg/ml. The qnrA gene was not detected in any of the quinolone-resistant isolates. Moreover, 23.6% (n=9) and 5.2% (n=2) of the quinolones-resistant isolates contained the qnrB and qnrS genes, respectively.
      Conclusion: Although 38 isolates were ciprofloxacin-resistant, the qnrB, qnrS genes were detected in a small number of isolates. This indicates the involvement of factors other than the qnr genes in resistance of these isolates to quinolones.
       Keywords: Klebsiella Pneumoniae, Qnr protein, Borujerd.

Seroepidemiological Study of Toxoplasma gondii in Patients with Multiple Sclerosis in Ahvaz, Southeastern Iran

Mehdi Tavalla

ABSTRACT
       Background and Objective: Toxoplasmosis is a common parasitic infection worldwide. The infection can be caused via consumption of contaminated meat and mother-to-child (congenital) transmission, causing changes in central nervous system tissue, eye irritation and sometimes death. The human form of the disease is often asymptomatic and may be accompanied with general discomfort and swelling of the lymph nodes when associated with chorioretinitis. Acute infection in immunocompromised individuals could lead to mortality. The purpose of this study was to determine the prevalence of anti-Toxoplasma gondii antibodies in serum of patients with multiple sclerosis (MS) referred to the MS Center in city of Ahvaz, southeastern Iran.
        Methods: Blood samples were taken from 100 patients with MS and 100 healthy control participants. After separating the serum, presence of anti-Toxoplasma antibodies (IgG, IgM) was evaluated by enzyme-linked immunosorbent assay.
        Results: Frequency of anti-Toxoplasma IgG was 38% and 21% in the patients and controls, respectively. Toxoplasma IgM antibodies were not detected in any of the study groups. Pearson correlation coefficient showed a significant association between Toxoplasma antibodies and MS.
       Conclusion: Due to high prevalence of toxoplasmosis in MS patients, it is recommended to measure serum titers of the patients regularly, and placed them under antiparasitic therapy when necessary.
      Keywords: Toxoplasma, Multiple sclerosis, MS, Ahvaz.

Evaluation of miR-101 Level in Patients with Chronic Hepatitis B Virus Infection and Liver Cirrhosis

Abdolvahab moradi

ABSTRACT
       Background and Objective: MiRNAs are small RNAs that are expressed in most eukaryotes, and can regulate gene expression by attaching to the 3’ end of target mRNA. MicroRNA-101 (miR-101) post-transcriptional regulation is important for host-virus interactions. In addition, miR-101 has a tumor suppressive role in liver cancer and metastasis, and induces apoptosis in tumor cells. We examined miR-101 expression in patients with chronic hepatitis B, hepatitis B virus (HBV)-associated cirrhosis and healthy individuals.
       Methods: The study was performed on 108 whole blood samples (36 samples from each group) collected in EDTA tubes. RNA was extraction by RNX-plus kit according to the manufacturer’s protocol. Finally, miRNA expression was evaluated using relative real time PCR.
         Results: A 2.4-fold increase was observed in miR-101 expression in patients with chronic hepatitis B, while there was a 3.5-fold increase in miR-101 expression in patients with HBV-associated cirrhosis compared with healthy controls (P=0.003). MiR-101 overexpression in patients with HBV-associated cirrhosis was more notable that in patients with chronic hepatitis B.
         Conclusion: According to the results, evaluating miR-101 expression may predict disease progression from chronic hepatitis B to HBV-associated cirrhosis.
         Keywords: MicroRNAs, Chronic Hepatitis B, Liver Cirrhosis, MiR-101.

Effects of Methanolic Extract of Ficus carica Leaves on Cystic Echinococcosis

Jaber Ali poor

ABSTRACT
          Background and objective: Surgery is the gold standard treatment for hydatidosis. Scolicidal agents could be used during surgery to kill protoscoleces and prevent cyst recurrence after rupturing of main lesion. Non-chemical agents with sufficient protoscolex-killing activity are known to be fully effective in this regard with fewer side effects. Fig tree is an Iranian native plant, which has been used for medicinal purposes in traditional medicine.
          Methods: After obtaining infected hydatid cyst of the liver from a slaughterhouse in Babol (Iran), the percentage of live protoscoleces was calculated by critical staining with 0.1 % eosin. Then, the protoscolex-killing activity of methanolic extract of fig leaves was evaluated at concentrations of 2.5, 5, 10, 15 and 20 mg/ml in 2, 4, 8, 12, and 16 minutes exposure times . Statistical analysis was performed using SPSS software (version 22).
          Results: Concentration of 20 mg/ml of the extract showed 100% protoscolex-killing activity within the first 2 minutes of exposure. In addition, the lowest protoscolex-killing activity (45%) was observed at concentration of 2.5 mg/ml after 2 minutes. The results also showed that the protoscolex-killing activity of the extract increases significantly in a concentration- and time-dependent manner (P <0.001).
         Conclusion: The methanolic extract of fig leaf at concentration of 20 mg/ml could exert significant scolicidal effect within 2 minutes of exposure. Therefore, complementary experiments should be performed on animal models to further assess the efficiency of the extract for killing protoscoleces of hydatid cyst during surgery.
          Keywords: Echinococcus granulosus, methanolic extract of Ficus carica, broth dilution.

Isolation and Characterization of Hyperthermophilic Nanobacteria from a Hot Spring in Ardabil, Iran

Reza Habibipour

ABSTRACT
       Background and Objective: Nanobacteria are nanometer-scale particles with different shapes, which have been a subject of debate in modern microbiology. They belong to a proposed class of living organisms, specifically cell-walled microorganisms with a size much smaller than the generally accepted lower limit for life. Since some microorganisms are able to continue growth at high temperatures, we aimed to isolate thermophilic bacteria from Gheinarcheh hot spring in Ardabil (Iran) and identify the characteristics of these microorganisms.
       Methods: Microbial mats were found in cultures from Gheinarcheh hot spring in North West of Iran. Synthetic media were prepared and used for isolation of protease-producing thermophilic bacteria, and identification of the features of microbial mats. Cultures were incubated at temperature range of 60-120 oC. Scanning electron microscopy, DNA extraction and polymerase chain reaction were used to further identify characteristics of the microbial biofilms.
        Results: Microbial biofilms of nanoparticles were detected in our samples. Growth of the bacteria increased at all temperatures tested. Results of scanning electron microscopy showed nano-sized particles in the scale of 60 nm. No band was visible in gel electrophoresis of polymerase chain reaction products.
Conclusion: This study is the first to report the presence of hyperthermophilic nanobacteria in Iran.
      Keywords: Nanobacteria, Hyperthermophile, Microbial Mat.

Detection of Virulence Genes Tcpa and Ctxb in Isolates from Surface Water and Salt Water Samples in Golestan, Iran

Shaghayegh Anvari

ABSTRACT
         Background and Objective: Vibrio is a genus of bacteria that are widely distributed in aquatic environments. The genus includes several important pathogens that endanger farm animals and humans who ingest seafood or water contaminated with the bacteria. Virulence of Vibrio spp. is regulated by ctxAB and tcpA genes. The aim of this study was to determine the prevalence of Vibrio spp., and tcpA and ctxB virulence genes in isolates from surface water and salt water samples collected from Golestan Province, Iran.
          Methods: Overall, 115 water samples were collected from the Caspian Sea coast, lagoons and rivers in the Golestan Province. The samples were filtered by membrane filtration method, and enriched in alkaline peptone water with 1% NaCl. The isolates were grown on TCBS agar, and identified by biochemical tests. Presence of the tcpA and ctxB virulence genes was investigated by polymerase chain reaction.
          Results: In this study, Vibrio alginolyticus was the predominant species (38%) isolated from the seawater and surface water samples, followed by Vibrio parahaemolyticus (23%), Vibrio harvei (15%), Vibrio fluvialis (14%) and Vibrio damsela (10%). The virulence genes were not detected in any of the isolates found in the study.
        Conclusion: This study indicates that V. alginolyticus is the most prevalent Vibrio spp. in surface water and seawater samples collected from the Golestan Province, Iran.
          Keywords: Environmental Vibrio, Surface water, ctx B gene, tcpA gene.

CCL2 Polymorphism in Drug-Resistant and Drug-Responsive Patients with Epilepsy in Isfahan, Iran

Zahra Rahimi

ABSTRACT
        Background and objective: Approximately 50 million people worldwide (1% of the world's population) suffer from epilepsy. Among 700 thousand people with epilepsy in Iran, 20% have refractory epilepsy. Accumulation of leukocytes in patients' brain parenchyma is thought to be related to different types of epilepsy. Recent clinical observations suggest that therapeutic strategies that interfere with leukocytes or cause them to migrate may have therapeutic efficacy in epilepsy. The aim of this study was to identify treatment-resistant patients, and investigate the association between polymorphism rs1024611 in CCL2 gene and drug resistance in patients with epilepsy in Isfahan, Iran.
        Methods: Blood samples were taken from 50 patients with intractable epilepsy (case group) and 50 drug-responsive patients with epilepsy (control group). Genomic DNA was extracted from peripheral blood by salting out method. Specific primers were designed by Oligo 7 software to investigate polymorphism rs1024611 using PCR-RFLP. The preliminary results for a number of samples were confirmed by sequencing.
        Results: The results of this study showed that there was a significant relationship between intractable epilepsy and presence of C allele.
       Conclusion: Similar to previous study, we found a significant association between CCL2 gene polymorphism and drug-resistant epilepsy.
        Keywords: Epilepsy, Drug Resistance, Polymorphism, CCL2.

Profile of Eight Prophage Sequences Present in the Genomes of Different Acinetobacter baumannii Strains

Majid Bouzari

ABSTRACT
          Background and Objective: Prophage sequences are major contributors to interstrain variations within the same bacterial species. Acinetobacter baumannii is a gram-negative bacterium that causes a wide range of nosocomial infections, especially in intensive care unit inpatients. Prophage sequences constitute a considerable proportion of several sequenced complete genomes of A. baumannii. The aim of this study was to investigate the presence of prophage sequences in A. baumannii strains isolated from burn patients, and compare the results with other studies.
          Methods: Presence of eight prophage sequences was investigated in the genome of ten multi-drug resistant A. baumannii isolates obtained from burn sites of 10 burn patients in a hospital in Isfahan, Iran. PCR and sequencing were performed to detect the prophage sequences. The presence of the eight prophage sequences in the genome of A. baumannii strains from other studies was investigated by BLAST analysis of whole nucleotide sequence of prophage sequences.
          Results: The isolates in the present study had different prophage sequence profiles. Two isolates did not contain any of the sequences, while two isolates contained three and two of the prophage sequences. Other isolates contained only one sequence. The prophage sequence profiles observed in this study were not found in A. baumannii isolates from other studies.
          Conclusion: The results of this study indicate that the prophage sequences profile can be useful for studying the epidemiology of A. baumannii strains.
          Keywords: Acinetobacter baumannii, genome, prophage sequences.