فهرست مطالب

Iranian Journal of Microbiology
Volume:5 Issue: 4, Dec 2013

  • تاریخ انتشار: 1392/09/05
  • تعداد عناوین: 21
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  • Saeed Shoja, Mojtaba Moosavian, Amir Peymani, Mohammad Amin Tabatabaiefar, Soodabeh Rostami, Nasim Ebrahimi Pages 315-322
    Background And Objectives
    Carbapenem resistant A. baumannii is an emerging cause of nosocomial infections. The aims of this study were identification of the most prevalent of carbapenem resistant genes, molecular typing and antimicrobial evaluation of A.baumannii in intensive care units.
    Materials And Methods
    Two hundred and six A. baumannii were isolated from tracheal tube discharge of hospitalized patients at different intensive care units in Ahvaz, Iran. Antimicrobial susceptibility test was done on all isolates. Multiplex and singleplex PCR were performed for detection of blaOXA-23-like, blaOXA-24-like, blaOXA-51-like,blaOXA-58-like, blaVIM, blaIMP, blaSPM and blaNDM genes. Genetic relationship of all isolates was determined by REP-PCR method.
    Results
    Out of 206 examined isolates, 198 (96.1%) isolates were resistant to imipenem and meropenem. However 3.9% isolates were sensitive to these antibiotics. The blaOXA-23-like and blaOXA-24-like genes were detected in 85% and 8.7% of strains, respectively. No blaOXA-58- like, blaIMP, blaVIM, blaSPM and blaNDM were detected. REP-PCR results showed that isolates were belonged to five genotypes: Genotype A was the most prevalent (P- value < 0.001): it was observed in 75 of 206 strains (36.4%). Genotype B, and C were found in 28.6% and 27.7%, respectively. The rate of other genotypes was as follows: D (2.4%), E (1%).
    Conclusion
    Based on the obtained results, the rate of carbapenem resistance was high among of A. baumannii which was isolated from intensive care units patients and oxacillinase genes were the most prevalent carbapenem resistant genes. These results revealed that three clones, A, B and C of A.baumannii are common in our hospitals.
    Keywords: Acinetobacter baumannii, Carbapenem resistant, Intensive Care Units
  • Shashidhar Vishwanath, Kiran Chawla, Anusha Gopinathan Pages 323-327
    Background
    Lower respiratory tract infections are among important causes of morbidity and mortality for all age groups. The emergence of multidrug resistant Gram-negative bacilli is an issue of increasing concern.
    Materials And Methods
    A retrospective study including respiratory specimens (sputum and BAL) was conducted in our tertiary care centre. Samples were processed for microscopy, culture and susceptibility testing following standard methods. Multidrug resistant Gram-negative bacilli causing lower respiratory tract infections were studied for their causation of disease. The effect of appropriate treatment on clinical outcome was observed.
    Results
    A total of 472 Gram-negative pathogens were isolated from sputum and broncho-alveolar lavage fluid specimens during the study period. Among these Gram-negative pathogens 175 (37%) were found to be multidrug resistant. Klebsiella pneumoniae 85 (48.6%) and Acinetobacter spp. 59 (33.7%) were the predominant multidrug resistant Gram-negative bacilli isolated. Based on clinico-microbiological correlation, 138 (78.9%) multidrug resistant isolates were found to be pathogenic and the rest 37 (21.1%) were considered as colonizers. After initiating appropriate antibiotic therapy, clinical improvement was seen in 110 (79.7%) patients. In the patients who showed improvement, amikacin (34.3%) and cefoperazone-sulbactum (21.8%) were found to be the most effective drugs.
    Conclusion
    A large majority of the isolated multidrug resistant Gram-negative bacilli were found to be pathogenic. Regular surveillance which directs appropriate empirical therapy; and good clinic-microbiological workup of each case of lower respiratory tract infection can reduce the morbidity and mortality associated with multidrug resistant organisms.
    Keywords: Acinetobacter spp., Gram, negative bacilli, Klebsiella pneumoniae, Lower respiratory infection, Multidrug resistance
  • Neda Mostatabi, Shohreh Farshad, Reza Ranjbar Pages 328-333
    Background And Objective
    Nowadays, the presence of extended-spectrum β-lactamases (ESBLs) producing strains in Serratia genus causes the emergence of resistance to many antibiotics. So, the lack of proper diagnosis of ESBLs strains can lead to failure in the treatment. The objective of the present study was to investigate ESBLs production in Serratia strains isolated from the clinical blood samples in Shiraz, Iran.
    Materials And Methods
    In this study, 39 Serratia strains isolated from the patients referred to Namazi Hospital, during a 2 year period were tested. The antimicrobial resistance of the isolates to 21 antibiotics was evaluated using Kirby-Bauer disk diffusion method. Combination disk method was used to determine the ESBL phenotype among the isolates. PCR was performed to investigate the presence of ESBL genes of SHV, OXA and TEM types.
    Results
    The lowest antibiotic resistance rates belonged to meropenem (7.69%) and imipenem (5.12%). Overall, positive ESBL phenotype was identified in 69% (n = 27) of the isolates, 70.37% (n = 19) for S. marcescens and 29.62% (n = 8) for S. liquefaciens. Results obtained by PCR showed that only 20.51% carried OXA gene and 15.38% carried SHV-1 gene. TEM gene was detected in none of the isolates.
    Conclusion
    This study showed a high prevalence of the emerging ESBL producing strains among clinical isolates of Serratia that could lead to an increase in antibiotic resistance. However, ESBLs genes other than those tested here may be more responsible for the emergence of ESBL phenotype among Serratia clinical isolates in our region.
    Keywords: Serratia, Extended, spectrum β, lactamases, OXA, SHV, TEM
  • Salman Khan, Priti Singh, Ashish Asthana, Mukhtar Ansari Pages 334-338
    Background And Objectives
    Shigella plays an important role as a causative organism of acute gastroenteritis, in children and others. Rapid emergence of antibiotic resistance warrants continuous monitoring of susceptibility pattern of bacterial isolates. We report here our findings about Shigella spp. isolates and their drug resistance patterns in Nepalese patients.
    Materials And Methods
    The study was conducted on 507 Nepalese patients with acute gastroenteritis attending outpatient and inpatient departments of Nepalgunj Medical college and teaching Hospital, Banke, Nepal from September 2011 to April 2013. Stool specimens were processed for isolation and identification of Shigella species following the standard microbiological methods while the disc diffusion test was used to determine antimicrobial resistance patterns of the recovered isolates at the central Laboratory of Microbiology.
    Results
    Sixty nine isolates were identified as Shigella species. S. flexneri, S. dysenteriae, S. boydii and S. sonnei accounted, respectively, for 42.03%, 27.54%, 21.74% and 8.70% of the total number of Shigella isolates. Resistance to nalidixic acid (95.65%), ampicillin (85.51%), co-trimoxazole (82.61%) and ciprofloxacin (47.83%) was observed. Among 69 isolates, 29 (42.03%) were from children aged 1-10 years and this group was statistically significant (P < 0.05), compared to the other age groups.
    Conclusions
    The study revealed endemicity of shigellosis with S. flexneri as the predominant serogroup in Nepalese patients. Children were at a higher risk of severe shigellosis. Nalidixic acid, ampicillin, co-trimoxazole and ciprofloxacin should not be used empirically as the first line drugs in treatment of shigellosis. Continuous local monitoring of resistance patterns is necessary for the appropriate selection of empirical antimicrobial therapy.
    Keywords: gastroenteritis, Shigella, Antimicrobial resistance, Nepal
  • Ramin Dibaj, Mohammad Mehdi Aslani, Parvin Heidarieh, Abdolrazagh Hashemi, Behrooz Ataei, Abbas Daei-Naser, Hasan Shojaei Pages 339-344
    Background And Objective
    Over 165 million cases of shigellosis occur in the world each year, mostly in developing countries. Outbreaks of shigellosis are associated with poor sanitation, natural calamities, contaminated food and crowded living conditions. In late summer 2006, during the final stage of an outbreak of shigellosis at a vast region of Isfahan province, Naein & Ardestan, our laboratory was assigned to investigate the outbreak in order to determine the causative agent.
    Materials And Methods
    A total of 146 rectal swabs which had been collected from the patients by local laboratories on separate days were screened using a battery of conventional and molecular tests.
    Results
    Thirteen specimens tested positive for Shigella spp. They were identified as S. sonnei (6, 46.1%), S. dysenteriae (4, 30.8%), S. flexneri (2, 15.4%) and Shigella spp (1, 7.7%) by conventional and molecular microbiological tests. According to ribotyping results the isolates were grouped into 3 distinct clusters encompassing the majority of isolates and a single line of descent representing isolate S122 which was nonreactive with any Shigella polyvalent antisera.
    Conclusion
    This diarrheal outbreak appeared to be the result of shigellosis. Despite the fact that Shigella sonnei was the predominant organism isolated from patients, the causative agent of outbreak diarrhea remains obscure, since other Shigella species were also involved. The serologic testing supports this conclusion, as do the molecular patterns of the Shigella isolates. Having considered the time of investigation which was in the late stage of the outbreak, it was very likely that a collection of endemic and epidemic clinical samples was screened resulting in isolation of various Shigella species.
    Keywords: Outbreak, Shigella, Ribotyping
  • Mohammad Mohammadzadeh, Mana Oloomi, Saeid Bouzari Pages 345-349
    Background And Objectives
    Enteropathogenic Escherichia coli (EPEC) divided into two groups typical and atypical (aspect). The main virulence genes are located in a pathogenicity island called LEE (Locus of Enterocyte Effacement). LEE frequently inserted in tRNA genes of selC, pheU and pheV in the bacterial chromosome. tEPEC and aEPEC strains have some differences in their pathogenicity. The purpose of this was to investigate the possible differences between tEPEC and aEPEC strains according to the virulence genes encoding by LEE and their relation to insertion sites.
    Materials And Methods
    In this study 130 E. coli isolates confirmed by biochemical analysis from diarrheal patients, were evaluated for EPEC pathotype by PCR. All EPEC strains tested for presence of some LEE encoded virulence genes and sites of LEE insertion by PCR method.
    Results
    Among 50 strains of EPEC 28 (56%) and 22 (44%) were typical and atypical strains respectively. 19 strains (30%) showed insertion in selC, 7 (14%) in pheU, 4 (8%) in pheV, 8 (16%) in pheU and pheV, 1 (2%) in selC and pheU, 6 (12%) in pheV, pheU and selC and 5 (10%) had no insertion in these sites. Moreover, spa (n = 8, 16%), espB (n = 16, 32%), espD (n = 18, 36%), espF (n = 8, 16%), espG (n = 13, 26%), espH (n = 12, 24%), map (n = 11, 32%) and tir (n = 4, 8%) were present among the strains.
    Conclusion
    Results showed that most of the virulence genes are present in tEPEC isolates. However, aEPEC isolates may acquire other virulence factors. The majority of tEPEC strains showed insertion at selC and aEPEC strains in pheV and pheU.
    Keywords: EPEC, LEE, virulence genes, tRNA
  • Mana Oloomi, Saeid Bouzari, Maryam Imani, Narges Akhtarian Pages 350-355
    Background And Objectives
    Baculovirus can be used as a vector in gene delivery system. Viral envelope of baculovirus would display expressed protein/peptide and it could render as a potential vaccine delivery system. In this regard, the gene coding for A subunit of shiga toxin (StxA) from Escherichia coli (E. coli) strain was cloned in a baculovirus expression system. StxA subunit has the ability to inhibit protein synthesis and this ability applied in cancer therapy. In this study, expression of StxA in baculovirus as a protein delivery system was assessed in vitro.
    Material And Methods
    StxA gene was cloned in pTriEx™ multisystem expression vector. This vector enables the protein expression in multisystem, E. coli and baculovirus. This construct was used to express the gene in E. coli and baculovirus. The construct containing StxA gene was made in baculovirus and expression was confirmed, then baculovirus expressing STXA transfect HeLa cells.
    Results
    The expression of STXA peptide (32kDa) was confirmed by SDS-PAGE and western blotting in both expression systems. The A subunit challenge to human cell Lines was applied as a delivery system by baculoviruses. On the other hand, the inhibition of cell proliferation was also demonstrated by baculovirus containing STXA subunit.
    Conclusion
    STXA peptide expression in baculovirus was shown in E. coli and baculovirus expression system. Furthermore, it was shown that A subunit of Shiga toxin delivered by baculovirus can inhibit cell proliferation in HeLa cells and leading to cell death. Therefore, this prototype system could be a promising model for in vivo cancer therapy and targeted protein delivery system.
    Keywords: Baculovirus, STXA peptide, Expression
  • Sanaz Goljanian Tabrizi, Javad Hamedi, Fatemeh Mohammadipanah Pages 356-365
    Background And Objectives
    Annual incidence of infection with S. Typhi is estimated to be about 17 million cases worldwide. A systematic search among actinomycete isolates from soil of Iran aimed at finding active actinomycetes against the causative agent of typhoid fever, Salmonella Typhi was carried out during this study.
    Materials And Methods
    Our anti-Salmonella screening program resulted in nine highly active actinomycete isolates. All nine antibiotic producing strains showed broad-spectrum antibacterial activity, as five strains showed antifungal activity as well. Based on microscopic morphology and cell wall analysis, all nine active actinomycete strains were representatives of the genus Streptomyces. Three of the producing strains including the isolates HG87, HG116 and HG443 with inhibition zone of >20 mm, were selected for further identification and investigation of cytotoxic effects.Results and
    Conclusion
    None of the producing strains showed cytotoxicity on HEK and USSC cell lines, while strain HG116 showed excellent antitumor activity on T47D cancer cell lines. Isolates HG87, HG116 and HG443 can be distinguished from the related species by some phenotypic and biochemical characteristics. Our results demonstrate the broad-range biological activity exhibited by bioactive compounds of soil actinomycetes from Iran.
    Keywords: Actinomycetes, Antibiotic, Cytotoxic activity, Salmonella enterica serovar Typhi
  • Mohadese Mozafari, Parissa Farnia, Mona Afraei, Zahra Derakhshani Nezhad, Mohammad Reza Masjedi, Ali Akbar Velayati Pages 366-373
    Background And Objectives
    Molecular epidemiology tools are widely used in determining epidemiology of tuberculosis. Spoligotyping is a molecular epidemiology method that is used for characterization and typing of Mycobacterium tuberculosis complex strains. The method is based on polymorphism of the chromosomal DR locus consisting of identical 36-bp DRs alternating with 35-41 unique spacers. The objective of this study was to investigate the prevalence of M. tuberculosis spoligotypes in different provinces of Iran.
    Materials And Methods
    1242 M. tuberculosis strains were isolated from TB patients of Mycobacteriology Research center (MRC). DNA was extracted from patient’s clinical samples. PCR was performed by using of specific primers for DR region. The amplified DNA was hybridized to the spoligotyping Membrane. Hybridized DNA was detected with ECL detection kit and by exposing ECL Hyperfilm to the membrane. The obtained result was entered to a binary format and was analyzed using SpolDB4 database.
    Results
    Spoligotyping resulted in 136 different patterns. Out of 1242 M. tuberculosis strains, 1165 strains (93.8%) were classified into 59 clusters and the remaining strains (6.2 %) were singleton.
    Conclusions
    The results of present study showed that strains of CAS family were more prevalent than other strains in Iran. Other prevalent families were Haarlem, T and Beijing, respectively.
    Keywords: Molecular epidemiology, Tuberculosis, Spoligotyping
  • Shima Mahmoudi, Setareh Mamishi, Mona Ghazi, Reihaneh Hosseinpour Sadeghi, Babak Pourakbari Pages 374-378
    Background And Objectives
    ESAT-6 (6-kDaearly secretory antigenic target) and CFP-10 (10-kDa culture filtrate protein) have been described as dominant antigens recognized by T-cells and considered as virulence factors in Mycobacterium tuberculosis. The aim of this study was to clone, express and purify recombinant ESAT-6 and CFP-10 proteins of M. tuberculosis in soluble form.
    Materials And Methods
    ESAT-6 andCFP-10 genes were amplified by PCR, cloned into pET32a (+) vector, and overexpress-ed using isopropyl-beta-D-thiogalactopyranoside in E. coli BL21 (DE3). ESAT-6 andCFP-10 proteins were purified by Ni-NTA affinity chromatography and were detected by anti- ESAT-6 and anti -CFP10 antibodies.
    Results
    ESAT-6 andCFP-10 genes were successfully expressed and purified. Anti- ESAT-6 and anti-CFP-10 antibodies were produced after induction of immunization against purified ESAT-6 andCFP-10 proteins in rabbit.
    Conclusion
    In this study, we cloned, expressed and purified sufficient amounts of ESAT-6 andCFP-10 and it would be tested for the development of diagnostic kit for M. tuberculosis in future.
    Keywords: M. tuberculosis, ESAT, 6, CFP, 10
  • Mansour Mayahi, Nader Mosavari, Saleh Esmaeilzadeh, Kaveh Parvandar Asadollahi Pages 379-382
    Background And Objectives
    Diagnosis of avian tuberculosis by conventional culture method is still considered as the “gold standard” technique. The main objective of this study was to compare growth of Mycobacterium avium subsp. avium on four specific Mycobacterial cultures such as glycerinated Lowenstein-Jensen medium, pyruvate-enriched Lowenstein-Jensen medium, mycobactin J-supplemented Herrold-egg yolk medium and plain Herrold-egg yolk medium
    Materials And Methods
    Eighty out of more than 600 pigeons were selected based on their clinical signs and poor health conditions. The birds were numbered and their clinical signs were registered in the working sheets, and under standard condi-tion, euthanized, subjecting to necropsy examinations, followed by bacterial culture on four specific media for Mycobacterium avium subsp. avium, including glycerinated Lowenstein-Jensen (LJG) medium, pyruvate-enriched Lowenstein-Jensen medium (LJP), mycobactin J-supplemented Herrold-egg yolk medium and plain Herrold-egg yolk medium.
    Results
    Fifty one Mycobacterium avium subsp. avium were isolated from pigeons. Mycobactin J-supplemented Herrold-egg yolk media yielded greater number of colonies in shorter incubation time in compare with other media.
    Conclusion
    It was concluded that most of the isolates need mycobactin as a growth factor.
    Keywords: Mycobacterium avium, pigeon, culture, Avium tuberculosis
  • Abbas Ali Imani Fooladi, Yousef Tarvedi Zadeh, Rezvan Mehrab, Raheleh Halabiyan, Taghi Azizi Pages 383-390
    Background And Objectives
    Tuberculosis (TB) is the oldest known bacterial disease in humans. Due to the rise of morbidity in recent years, early diagnosis of the disease is necessary.
    Materials And Methods
    In this study we used Fluorescent Amplification–Based Specific Hybridization (FLASH) PCR to targetIS6110 for rapid detection of M. tuberculosis (MTB). To investigate the important factors influencing the risk of TB, data from patients and their medical records were analyzed.
    Result
    The sensitivity and specificity of FLASH-PCR for detecting MTB were determined as 93.33% and 92.5%, respectively. The findings of this study have suggested that removal of the contaminants in FLASH-PCR sign ificantly reduced the detection time, and MTB was much more rapidly detected in the clinical specimens compared to the conventional culture and smear examination. Results of the medical survey showed that the majority of TB patients were males, over 51 years old, smokers, with pulmonary TB and normal chest X-ray (CXR).
    Conclusion
    MTB can be rapidly detected inclinical specimens using FLASH-PCR in comparison with culture and smear examination.
    Keywords: Mycobacterium tuberculosis, FLASH, PCR, Rapid detection, Tuberculosis
  • Mohan Kumar Pasupuleti, Ravindra Reddy Nagireddy, Roopa Dinahalli, Deepa Anumala, Avula Kishore Kumar, Vinay Chavan Pages 391-395
    Background And Objectives
    Gingival and periodontal diseases are associated with specific bacterial infections. The main aim of the study was to know whether the periodontitis is associated with an increased risk for acute myocardial infarction (AMI) and to know the distribution of Porphyromonas gingivalis in patients with acute myocardial infarction associated with chronic periodontitis and acute myocardial infarction Groups.
    Materials And Methods
    Out of 50 patients, 20 were diagnosed as acute myocardial infarction associated with chronic periodontitis (Group I), twenty patients were suffering from AMI (Group II) and 10 patients were healthy (control Group III).Results and
    Conclusion
    Periodontal pathogens were identified by phenotypic, enzymatic and hybridization methods. The total bacterial load and the number of Porphyromonas gingivalis pathogens were more in Group I when compared to Group II and Group III. Thus, the present study confirmed an association between periodontitis and AMI.
    Keywords: Acute myocardial infarction, periodontal disease, Porphyromonas gingivalis
  • Mohammad Khalifeh Gholi, Fereshteh Shamsipour, Hossein Ajhdarkosh, Naser Ebrahimi Daryani, Mohammad Reza Pourmand, Mostafa Hosseini, Amir Ghasemi, Mohammad Hasan Shirazi Pages 396-401
    Background And Objectives
    Invasive and non-invasive techniques are used to diagnose H. pylori infection. Some factors influence the choice of a diagnostic test, such as the sensitivity and specificity of the tests, the clinical circumstances and the cost-effectiveness of the testing strategy. The aim of this study was to reveal the relationship between different H. pylori infection diagnosis methods, and clarify the application scope of each diagnosis method.
    Materials And Methods
    91 patients were included in the study, and specimens including biopsies, blood and stool were taken. Biopsies were evaluated by hematoxylin and eosin, and Giemsa staining. A sequence of 294 bp in the ureC (glmM) gene was amplified. The rapid urease test (RUT) was performed using a non-commercial validated test. Stool samples were analyzed using a polyclonal ELISA stool antigen test. A serological assay for IgG antibodies was performed by a commercial Helicobacter pylori IgG ELISA kit.
    Results
    According to the predefined criteria, a total of 46 (50.5%) patients tested were positive by at least 2 of the 3 biopsy-based methods. The best sensitivity (95.6%) belonged to histology and RUT. The sensitivities of other tests including PCR, serology and stool antigen test were 93.5%, 91.3% and 73.9%, respectively. RUT showed the best specificity (100%), and the specificities of the other tests, including PCR, stool antigen test, histology and serology, were 95.6%, 86.7%, 77.8% and 55.6%, respectively.
    Conclusion
    In view of the better results obtained for invasive vs. non-invasive tests, for a more accurate diagnosis, it is advisable not to solely rely on non-invasive methods of H. Pylori diagnosis.
    Keywords: Helicobacter pylori, Histopathology, Stool antigen test, ELISA, RUT, PCR
  • Abdollah Derakhshandeh, Roya Firouzi, Ali Goudarztalejerdi Pages 402-405
    Background And Objectives
    Brucella, causative of brucellosis, has some potential virulence factors involved in Brucella replication and its strategies to circumvent the immune response. One of them is the virB gene that encodes the type ІV secretion system proteins (T4SS) involved in intracellular replication of organism. Brucella virulence factor A (bvfA), and urease (ure) has also been described as being implicated in survival, and virulence in the hosts. The aim of this study was to investigate the B. melitensis virulence factor genes among Brucella isolated from aborted fetuses of sheep and goats in Fars province, southern Iran.
    Materials And Methods
    A total of 42 isolates of B. melitensis isolated from aborted fetuses between 2005-2011 in Fars province of Iran was used in this study. PCR assay was performed in order to detect the virB, bvfA, and ure genes using specific primers.Results and
    Conclusions
    The frequency of bvfA, virB, and ure genes was 78.50%, 73.80%, and 88.09% among all isolates respectively. The results of the present study showed that most Brucella isolates from this region have virulence factors genes (virB, bvfA, ure) in their genome, and most B. melitensis had ure genes that has been hypothesized to play a role in the pathogenesis of disease.
    Keywords: Brucella melitensis, Virulence genes (bvfA, virB, ure), Sheep, Goat, Fars province, Iran
  • Parisa Badiee, Amir Arastehfar, Hadis Jafarian Pages 406-410
    Background And Objectives
    The aim of this study was to compare direct microscopic examination with culture and PCR for the diagnosis of Mucorales infection in blood and tissue specimens.
    Material And Methods
    Blood samples and tissue specimens were obtained from 28 patients (total 58 samples) with suspected invasive fungal infection and cultured on proper media. Direct smear of tissue samples was done with potassium hydroxide, hematoxylin and eosin, and methenamine silver staining. DNA extracted from blood and tissue specimens were used for semi-nested PCR targeting 18S rDNA of Mucorales species.
    Results
    Mucormycosis was documented in 7/28 (25%) of tissue specimens with positive findings by direct smear, of which PCR and culture were positive in 6 (86%) and 5 (70%) specimens, respectively. The etiologic agents were Mucor spp. and Rhizopus spp. However, culture and PCR results for all blood specimens were negative.
    Conclusions
    As the orders of Mucorales do not have well growth in culture media, PCR with tissue specimens is more sensitive than tissue or blood culture methods. Unfortunately, there is no alternative method for direct smear, which is an invasive method. Molecular methods may be helpful in these cases.
    Keywords: Invasive fungal infection, Mucoracea, Semi, nested PCR, 18S rDNA
  • Maryam Fatemeh Sheikholeslami, Javid Sadraei, Parissa Farnia, Mehdi Forozandeh, Hamid Emadi Kochak, Payam Tabarsi, Sayed Ali Reza Nadji, Majid Pirestani, Mohammad Reza Masjedi, Ali Akbar Velayati Pages 411-417
    Background And Objectives
    With increasing rate of immunodeficiency diseases in the world, opportunistic micro-organism such as Pneumocystis jirovecii (P. jirovecii) become more important. Little information is available on prevalence of this life-threatening microorganism in Iran. This study was designed to determine the colonization and the rate of active disease caused by P. jirovecii in two groups of Iranian immunosuppressed patients.
    Materials And Methods
    Two hundred and fifty five pulmonary samples were collected from two groups of immunosuppress-ed patients to detect a 260bp fragment of mt LSU rRNA gene of P. jirovecii by nested PCR. The first group was COPD patients consumed oral, inhaled or injectable corticosteroid and the second group was patients with malignancies under chemotherapy. Both groups were referred to National Research Institute of Tuberculosis and Lung Disease and Imam Khomeini hospital because of pulmonary symptoms. All patients introduced to this project were confirmed HIV sera-negative by ELISA and western blot test.
    Results
    The mean age of COPD patients was 66.5 ± 11 (41-88) years and all of them were men. The mean age of patients with malignancy (PMs) was 43 ± 11 (23-65) years and 51.6% were men. The P. jirovecii was colonized in 7 of 89 COPD patients (7.9%) and its DNA was isolated from 11 of 153 PMs (7.2%). The microorganism could cause active disease in 7 of 67 (10.5%) PMs who suffered from pneumonia.
    Conclusion
    The study showed that P. jirovecii was one of colonizing agents in the COPD patients, but it could cause active disease in PMs. Generally, the microorganism can exist in the lung of non-HIV+ immunosuppressed patients. Therefore, it should be considered as a potential infective agent in non-HIV+ immunocompromised patients.
    Keywords: Colonization, P. jirovecii, COPD, Non, HIV+ immunocompromised patients
  • Razieh Talebzadeh, Daryoush Alipour Pages 418-421
    Background And Objectives
    Essential oils may improve the utilization of nutrients by ruminal microorganisms. The aim of this study was to assess the effect of different doses of ajowan essential oils (AEO) on growth and fibrolytic activity of anaerobic fungi, and generic distribution of ciliated protozoa (in vitro).
    Material And Methods
    Different doses of AEO (0, 150, 300, 450 and 600 ppm) were added to experimental tubes. The effect of AEO was evaluated on growth and fibrolytic enzyme activity of an anaerobic fungus (Neocalimastix spp.). Generic distribution of ciliated protozoa were evaluated in response to different doses of AEO.Results and
    Conclusion
    The growth of fungus (Neocalimastix spp.) were inhibited and activity of fibrolytic enzymes of fungus were reduced by adding AEO. Also, an inhibitory effect was seen in concentration of ciliated protozoa and some genus were completely disappeared at the doses of 300 ppm and higher. The doses used in this study reduced the fibrolytic activity of the studied rumen microorganisms which is undesirable in practical animal nutrition. Further research is needed to assess the effects of AEO at lower doses on these parameters and also proteolysis and methanogenesis.
    Keywords: Rumen, Carum copticum, anaerobic fungi, ciliated protozoa, ajowan
  • Reyhaneh Noormohamadi, Fatemeh Tabandeh, Parvin Shariati, Maryam Otadi Pages 422-427
    Background And Objectives
    Lipases are valuable biocatalysts which are widely used in the detergent, food, dairy and pharmaceutical industries. The aims of the present study included the isolation of a lipase-producer from industrial zones and the partial characterization of the enzyme.
    Materials And Methods
    A number of bacteria were isolated from sites related to the oil industries. An isolate forming a halo zone in a selective medium (TW agar) was then selected and grown on a medium suitable for the production of lipase. The isolate was subsequently identified by the 16S rRNA sequencing method, and its enzyme activity was measured by a spectrophotometer using pNPP as a substrate.
    Results
    The selected isolate was identified by the molecular method as Pseudomonas sp. Its extracellular lipase activity was 41.5 ± 1.4 U/ml, and the high affinity of this enzyme for the substrate was indicated by the kinetic parameters of Km and Vm, which were estimated by the the Lineweaver-Burk plot as 0.77 mM and 49.5 U/ml, respectively. Activation energy of lipase calculated from the Arrhenius plot was found to be 20.78 kJ/mol, and a temperature coefficient (Q10) of 4.39 indicated the high catalytic activity of the enzyme and the temperature dependence of the enzymatic reaction.
    Conclusion
    The results demonstrated that the indigenous isolate could have potential applications in many relevant industries.
    Keywords: Lipase, Pseudomonas, Kinetic constants, Thermodynamic parameters
  • Darab Yazdani, Zainal Abidin Mior Ahmad, Tan Yee How, Indu Bala Jaganath, Sahar Shahnazi Pages 428-433
    Background And Objectives
    Food contamination by aflatoxins is an important food safety concern for agricultural products. In order to identify and develop novel antifungal agents, several plant extracts and isolated compounds have been evaluated for their bioactivities. Anti-infectious activity of Piper betle used in traditional medicine of Malaysia has been reported previously.
    Materials And Methods
    Crude methanol extract from P. betel powdered leaves was partitioned between chloroform and water. The fractions were tested against A. flavus UPMC 89, a strong aflatoxin producing strain. Inhibition of mycelial growth and aflatoxin biosynthesis were tested by disk diffusion and macrodillution techniques respectively. The presence of aflatoxin was determined by thin-layer chromatography (TLC) and fluorescence spectroscopy techniques using AFB1 standard. The chloroform soluble compounds were identified using HPLC-Tandem mass spectrometry technique.
    Results
    The results, evaluated by measuring the mycelial growth and quantification of aflatoxin B1(AFLB1) production in broth medium revealed that chloroform soluble compounds extract from P. betle dried leaves was able to block the aflatoxin biosynthesis pathway at concentration of 500μg/ml without a significant effect on mycelium growth. In analyzing of this effective fractions using HPLC-MS2 with ESI ionization technique, 11 phenolic compounds were identified.
    Conclusion
    The results showed that the certain phenolic compounds are able to decline the aflatoxin production in A. flavus with no significant effect on the fungus mycelia growth. The result also suggested P. betle could be used as potential antitoxin product.
    Keywords: phenolic compounds, A. flavus, aflatoxin biosynthesis, Piper betle, LC, MS2
  • Narjes Jafari, Rouha Kasra Kermanshai, Mohammad Reza Soudi Pages 434-440
    Background And Objectives
    Synthetic dyes are recalcitrant to degradation and toxic to different organisms. Decolorization of textile wastewaters is one of the major concerns since last decades. Physical-chemical treatments are very expensive and frequently producing large amounts of toxic wastes. Biological treatments can be more convenient. In the present study, an attempt has been made for decolorization of azo dyes using microbial process.
    Material And Methods
    Screening of microorganisms capable of azo dye decolorization was performed from activated sludge. The decolorization of various dyes (Reactive Black 5, Reactive Orange 16, Reactive Red 198, Direct Blue 71, Direct Yellow 12 and Direct Black 22) was determined by measuring the absorbance of culture supernatant at their λmax. Culture supernatants were also analyzed for UV-Vis absorption between 200-800 nm. The effect of aeration, temperature, different concentrations of glucose and NaCl was studied with an aim to determine the optimal conditions required for maximum decolorization.
    Results
    The yeast (strain JKS4) which had high ability to decolorize different azo dyes was isolated. Under aerobic condition, the yeast strain showed 85.7% of decolorization at 200 mg/l Reactive Black 5 (as a model azo dye), 1% (w/v) glucose concentration and 35°C after 24 h. All the examined dyes were extensively decolorized (53.35-97.9%) after 24 h. With elongated incubation period, complete decolorization was observed in presence of all dyes. From the physiological properties and phylogenetic analysis based on the 26S rDNA sequences, strain JKS4 was classified into Candida palmioleophila.
    Conclusions
    Because of high decolorizing activity against various azo dyes commonly used in the textile industries, it is proposed that the isolated yeast may have a practical application in the biotransformation of various dye effluents.
    Keywords: Azo dyes, Decolorization, Candida palmioleophaila, Textile Wastewater