فهرست مطالب

Jundishapur Journal of Microbiology
Volume:7 Issue: 9, Sep 2014

  • تاریخ انتشار: 1393/07/30
  • تعداد عناوین: 17
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  • Ali Fattahi Bafghi *, Mohammad Noorbala, Mohammad Taghi Noorbala, Mahdi Aghabagheri Page 1
    Background
    Cutaneous leishmaniasis (CL) is an endemic disease in developing countries. Although pentamidine orantimonite (Glucantime) has been recommended for cutaneous leishmaniasis treatment by the World Health Organization, there are some concerns too such as high cost, side effects, need for frequent injections, and restricted efficacy. Therefore, different methods have been used for CL treatment so far..
    Objectives
    This study assessed the sensitivity of two parasite agents of cutaneous leishmaniasis: Leishmania major and L. tropica to zinc sulphate in vitro. In the present study, the zinc sulphate effect on urban and rural strains of cutaneous leishmaniasis, viability of old world, in vitro is under investigation..
    Materials And Methods
    The design of the present study was experimental (laboratory-trial) based. Iranian endemic species of L. major and L. tropica were appropriately collected, proliferated, and maintained in the standard culture. Afterward, the proper concentrations of zinc sulphate were provided, sterilized, and added to the cultures containing parasites. In different intervals, parasites were counted by two
    Methods
    the slide and cell proliferation ELISA..
    Results
    Both parasite species showed sensitivity to zinc sulphate in vitro and in comparison with the control group, their numbers were reduced. Zinc sulphate (in concentrations of 0.5, 1, 2, and 3percent) was added to the cultures containing parasites, and the total number of the live parasites was counted through the slide method (Neubauer slide) every day up to the fifth day. The results were analyzed and found statistically significant (P < 0.05). In the second phase, the counting process was repeated with the addition of zinc sulphate compound with different concentrations (3, 4, 5, and 6 percent) and live parasite numbers were counted by ELISA method after 24 hours. The findings revealed that all the cultures containing zinc sulphate showed a slower growth in comparison to the control group. The mentioned difference was statistically significant (P < 0.05)...
    Conclusions
    Considering the safety of zinc sulphate compound in comparison with Glucantime, there is a possibility of using it in the treatment of CL caused by both species of L. major and L. tropica. It is obvious that more researches are mandatory both in vivo and in vitro to figure out its daily dosage, proper concentration, time and duration, and possible side effects..
    Keywords: Cutaneous Leishmaniasis, Zinc Sulphate, In Vitro, Therapeutics
  • Mahamoudreza Panahibazaz, Mojataba Moosavian, Gholamreza Khataminia, Mostafa Feghhi *, Farsim Yazdi, Effat Abbasi Montazeri Page 2
    Background
    Postoperative endophthalmitis is one the most serious complications of cataract surgery. The majority of causative organisms in this destructive infection come from the patient’s own periocular flora. Efforts have been made to reduce the virulence of organisms in the eyelid and conjunctiva with perioperative topical antibiotics, preparation of surgical field, covering eyelids and conjunctival surface with 5% povidone–iodine solution and intracameral antibiotics at the time of surgery to minimize the risk of endophthalmitis..
    Objectives
    We assessed the effect of subconjunctival injection of cefazolin and pouring povidone-iodine on the conjunctiva bacterial colony forming units (CFU) in phacoemulsification cataract surgery..Patients and
    Methods
    In this prospective, randomized, double-blind clinical trial, 122 patients having phacoemulsification cataract surgery with clear corneal incision and topical anesthesia were randomized into two groups including group 1 (subconjunctival injection of cefazolin) and group 2 (recipients of a drop of povidone-iodine). Cultures were collected from the bulbar conjunctiva at the injection site and from the corresponding location in the patient’s eye, three different times..
    Results
    The mean of eyelid samples on blood and chocolate agars, on the day after compared to the day before the surgery in group 1 showed a 52% and 56% reduction. These values were 58% and 50% in group 2 (P < 0.05). The mean CFU of conjunctiva before and at the end of surgery on blood and chocolate agars showed 57% and 56% reduction in group one and 51% and 52% reduction in group 2 (P < 0.05). While comparing mean CFU of conjunctiva at the end and one day post-surgery (interval of 14 ± 2 hours) showed 27% and 27% increase in group 1 and 20% and 21% increase in group 2 (P < 0.05), which reflects conjunctival flora proliferation during the early postoperative period..
    Conclusions
    Due to the good tolerance of patients towards topical anesthesia, pouring a drop of povidone-iodine 10% seems to be a simple and acceptable method to reduce the growth of microorganisms of the conjunctiva..
    Keywords: Injection, Cefazolin, Povidone, Iodine, Phacoemulsification
  • Mahshid Dastkhosh, Pooneh Rahimi *, Setareh Haghighat, Peyvand Biglari, Nader Howaizi, Reza Saghiri, Akram Roohandeh Page 3
    Background
    Rabies is a major zoonotic viral disease and is detected using the World Health Organization standard diagnostic techniques. Rabies detection is preferably done using the fluorescent antibody technique (FAT) that provides reliable diagnosis with almost 100% accuracy for all variant strains, if a proper conjugate is used. Rabies virus nucleoprotein (NP) is the most important protein used in production of a specific diagnostic conjugate..
    Objectives
    The aim of this study was to extract the cell-associated rabies virus NP from infected Baby Hamster Kidney cell clone (BSR) with rabies virus (Pasteur vaccine strain/PV) and purify for a future project to produce an anti-NP conjugate..
    Materials And Methods
    Pasteur vaccine strain (PV) as the standard rabies vaccine strain with a focus-forming dose (FFD) of 105 was inoculated in to the BSR cell culture at a concentration of 106 cells per milliliter. Infected cells were harvested 72 hours after infection and the rabies NP was extracted from these cells by low-speed centrifugation and purification by ultracentrifugation in cesium chloride (CsCl) gradient. For analysis, the purified NP was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)..
    Results
    The volume of the lysate was 15 mL and it became 2.5 mL after purification, with a concentration of 3.25 mg/mL. The corresponding band to the cell lysate protein on the SDS-PAGE had a molecular weight of 50 KDa, similar to the molecular weight of NP in rabies virus..
    Conclusions
    The rabies virus NP could be extracted and purified in an appropriate amount from infected cell culture. The results of SDS-PAGE analysis showed that the intact rabies virus NP had been purified properly and thus could be used for further steps to produce the specific diagnostic rabies conjugate..
    Keywords: Fluorescent Antibody Technique, Diagnosis, Rabies Virus, Isolation, Purification, Nucleoproteins
  • Gholam Hossein Ebrahimipour, Zahra Khosravibabadi *, Hossein Sadeghi, Atusa Aliahmadi Page 4
    Background
    Antibiotics are usually assumed as secondary metabolites produced during the idiophase of microbial growth, which can kill or inhibit the growth of other microorganisms. Nowadays, indiscriminate use of antibiotics has resulted in resistant microorganisms. Therefore, screening researches on products with antimicrobial activities are necessary..
    Objectives
    To find new antibiotics to defend against pathogenic microorganisms resistant to common antibiotics, the bacterium isolated from skin of the frog called Rana ridibunda was studied for its antimicrobial activities..
    Materials And Methods
    An antibiotic-producing bacterium was isolated from the frog skin. The bacterium was identified based on 16SrDNA sequencing and biochemical and morphological characteristics. Antimicrobial activity of the culture supernatant was examined against laboratorial standard bacteria by disc diffusion and minimum inhibitory concentration (MIC) methods. To characterize the produced antimicrobial compound, the culture supernatant of the bacterium was washed by chloroform and dried at 40°C; then, the antimicrobial substance was extracted by methanol and acetone and detected by bioautography on silica gel plates. Dialysis tube was used to find the molecular weight of this substance..
    Results
    The isolated bacterium was identified as a new strain of Bacillus atrophaeus. The antimicrobial substance exhibited heat stability between 25ºC and 100ºC and was active in a broad pH range from 2.0 to 11.0. The bioautography assay showed that methanol was the optimum solvent for the extraction of antimicrobial substance. The dialysis tube indicated that the antimicrobial substance weight was less than 1 kDa and the compound did not precipitate with ammonium sulfate..
    Conclusions
    This study showed that some properties of antimicrobial substances produced by the GA strain differed from other peptide antibiotics produced by the genus Bacillus such as bacitracin, which increases the likelihood of its novelty..
    Keywords: Antimicrobial Agent, Frog, Bacillus
  • Fatemeh Fotouhi *, Behrokh Farahmand, Behnaz Heidarchi, Maryam Esghaei, Sima Rafati, Masoumeh Tavassoti Kheiri Page 5
    Background
    Permanent antigenic variation of influenza viruses causes a major concern to develop an effective human influenza vaccine. Conserved antigens are new vaccine candidates because it is not necessary to match the prepared vaccine with circulating strains. Ion channel M2 protein is conserved among all influenza A viruses, allowing the virus to enter host cells..
    Objectives
    To prepare an effective vaccine against influenza A viruses, a chimerical DNA plasmid encoding Influenza virus M2 protein and Leishmania major HSP70 was constructed..
    Materials And Methods
    Influenza A/New Caledonia/20/99 (H1N1) was inoculated into MDCK cell line and total RNA was extracted. The full length M2 gene was amplified by RT-PCR using designed specific primers, cloned into pGEM-T Easy cloning vector and completely sequenced. The M2 gene was then subcloned into the pcDNA upstream of HSP70 gene. Recombinant plasmids were transfected into COS-7 cells to evaluate protein expression..
    Results
    The recombinant plasmids were confirmed by PCR, restriction enzyme analysis and sequencing. Three dimensional structure of chimer protein was assessed using specific software. Transient protein expression in eukaryotic cells was confirmed by specific mRNA detection, indirect Immunofluorescence test and western blotting..
    Conclusions
    M2-HSP70 chimer protein was successfully expressed in eukaryotic cells. Computational studies of chimer peptide sequence revealed that fusing HSP to the C-terminal of M2 protein does not mask the predominant epitope of M2. HSP70 is a molecular chaperon and immunostimulatory component. Genetically fusing antigens to HSPs leads to the enrichment of DNA vaccine potency. The immunogenicity of this construct with different formulation would be evaluated in further investigations..
    Keywords: Influenza Virus, Vaccine, Molecular Chaperone, Heat Shock Protein, M2 Protein
  • Jila Yavarian, Nastaran Gavvami, Setareh Mamishi * Page 6
    Background
    Encephalitis is swelling and inflammation of brain, usually due to viral infection. Viral encephalitis symptoms could be fever, headache, altered level of consciousness, and seizures..
    Objectives
    The aim of this study was detection of human herpesvirus-6 (HHV-6) DNA in cerebrospinal fluid (CSF) of patients with symptoms of possible acute encephalitis and without typical signs or symptoms of roseola infantum, using real-time polymerase chain reaction (PCR)..Patients and
    Methods
    We studied children two years old or younger, admitted to the pediatric emergency ward with encephalitis-like symptoms. Our evaluation included detection of HHV-6 in CSF of these patients. After DNA extraction, real-time PCR was performed with primers and a probe specific for the U22 open reading frame of both HHV-6A and B..
    Results
    From a total of 114 patients, HHV-6 was detected in 10 (8.8%), 90% of which were boys with mean age 7.7 months and median of 7.5 months. No significant differences were found in clinical presentations and laboratory findings between the patients positive and negative for HHV-6. All the children had complete recovery without neurological deficit or death..
    Conclusions
    According to this research and prevalence of HHV-6 in children, evaluation of CSF (detecting the HHV-6 DNA by PCR) is recommended in patients younger than 13 months with possible encephalitis..
    Keywords: Real, Time Polymerase Chain Reaction, Human Herpesvirus6, Cerebrospinal Fluid, Encephalitis
  • Pedram Haddadi, Soheila Zareifar, Parisa Badiee *, Abdolvahab Alborzi, Maral Mokhtari, Kamiar Zomorodian, Keyvan Pakshir, Hadis Jafarian Page 7
    Background
    Pediatric patients with neutropenia are vulnerable to invasive Candida infections. Candida is the primary cause of fungal infections, particularly in immunosuppressed patients. Candida albicans has been the most common etiologic agent of these infections, affecting 48% of patients.
    Objectives
    The aim of this study was to identify Candida spp. isolated from children with neutropenia and determine the antifungal susceptibility pattern of the isolated yeasts..Patients and
    Methods
    In this study 188 children with neutropenia were recruited, fungal surveillance cultures were carried out on nose, oropharynx, stool, and urine samples. Identification of Candida strains was performed using germ tube and chlamydospore production tests on an API 20 C AUX system. Susceptibility testing on seven antifungal agents was performed using the agar-based E-test method..
    Results
    A total of 229 yeasts were isolated. Among those, C. albicans was the most common species followed by C. krusei, C. parapsilosis, C. glabrata, C. tropicalis, C. famata, C. dubliniensis, C. kefyr, and other Candida species. C. glabrata was the most resistant isolated yeasts, which was 70% resistant to fluconazole and 50% to itraconazole, 7.5% to amphotericin B and 14% to ketoconazole. All the tested species were mostly sensitive to caspofungin..
    Conclusions
    Knowledge about the susceptibility patterns of colonized Candida spp. can be helpful for clinicians to manage pediatric patients with neutropenia. In this study, caspofungin was the most effective antifungal agent against the colonized Candida spp. followed by conventional amphotericin B..
    Keywords: Neutropenia, Candidemia, Antifungal, Susceptibility, Caspofungin, Amphotericin
  • Fahem Mohsin Mahmood, Haider Sabah Kadhim *, Liqaa Riadh Mousa Al Khuzaee Page 8
    Background
    Human papillomavirus (HPV) is the causal factor for cervical cancer. However, the role of HPV infection in ovarian cancer is unclear..
    Objectives
    This study aimed to determine the presence of human papillomavirus-16 (HPV-16) in ovarian tumor tissues..Patients and
    Methods
    This was a retrospective study, which included 61 Archived human ovarian tumor tissues embedded in paraffin blocks. The ovarian tumor tissues were divided into four groups. The first group was the malignant ovarian epithelial tumor group; it included 31 cases with invasive surface epithelial ovarian tumors. The second group was the borderline epithelial ovarian tumor group: it included four cases with borderline intermediate malignancy. The third group was the benign epithelial ovarian tumors group: it included 18 cases with benign epithelial ovarian tumors. The fourth group had functional ovarian cystic lesions: it included eight cases with non-neoplastic functional ovarian cysts. Sections were made from each of the paraffin embedded blocks and examined using immunohistochemistry to detect HPV 16-E6-oncoprotein in ovarian tumor tissues..
    Results
    Out of the eight cases with functional cysts only one case (12.5%) expressed HPV. No HPV expression was seen in cases with benign and borderline tumors. Out of the 31 cases with one malignant surface epithelial ovarian tumor only three (9.67%) cases expressed HPV. There was no significant statistical difference in HPV expression among neoplastic and non-neoplastic ovarian tumors included in the present study (P= 0.476)..
    Conclusions
    HPV type 16 was detected in only 9.67% of malignant epithelial tumors. It appears that HPV infection plays a relatively minor role in the pathogenesis of ovarian carcinomas..
    Keywords: Human Papillomavirus, 16, Tumor, Immunohistochemistry
  • Bahman Abdi-Hachesoo *, Rahem Khoshbakht, Hassan Sharifiyazdi, Mohammad Tabatabaei, Saeid Hosseinzadeh, Keramat Asasi Page 9
    Background
    Campylobacter is one of the leading bacterial species causing foodborne illnesses in humans. Antimicrobial agents have been extensively used for treatment of Campylobacter infections; but in the recent years, both animal and human isolates of this bacterium have shown resistance to several antibiotics such as tetracycline..
    Objectives
    The aim of this study was to investigate the presence of genetic determinants of tetracycline resistance in Campylobacter spp. recovered from poultry carcasses in Shiraz, Iran..
    Materials And Methods
    Eighty-three thermophilic Campylobacter spp. Isolates were first identified based on multiplex polymerase chain reaction (PCR) and then screened for presence of tetracycline resistance genes (tet (A), tet (B), tet (O) and te (S)) by PCR..
    Results
    The overall prevalence of Campylobacter jejuni and C. coli among the examined isolates was 51.8% and 48.2%, respectively. Tetracycline resistance genes of tet (B) and tet (S) were not seen among these Campylobacter spp. Isolates, whereas the most common tet gene identified was tet (O), found in 83.1% (69/83) of all the isolates. The tet (O) gene sequence comparison between C. jejuni and C. coli showed 100% similarity and these sequences (JX853721and JX853722) were also identical to the homologous sequences of other strains of Campylobacter spp. existing in the GenBank databases. In addition, tet (A) was found in 18% (15/83) of Campylobacter spp. isolates. To our knowledge, this represents the first report of tet (A) in Campylobacter spp. There was 100% homology between the sequences of tet (A) from this study (JX891463 and JX891464) and the tet (A) sequences mentioned for other bacteria in the GenBank databases..
    Conclusions
    The high prevalence of tet (O) resistance gene along with new detection of tet (A) resistance gene in Campylobacter spp. isolated from poultry carcasses revealed an extensive tetracycline resistance among Campylobacter isolates from poultry in Iran. It emphasized the need for cautious use of tetracycline in poultry production to decrease the extension of tetracycline-resistant Campylobacter spp..
    Keywords: Campylobacter, Tetracycline Resistance
  • Seyed Mohammad Hassan Aletayeb, Ashraf Sepehran, Hazhir Javaherizadeh * Page 10
    Introduction
    Neonatal parotitis is a rare disease. Neonatal suppurative parotitis commonly presents with facial swelling, irritability, tenderness of parotid region, and with or without fever. Acute neonatal suppurative parotitis is one of the differential diagnoses of facial swelling with a prevalence of 3.8/10’000 of neonatal admission..
    Case Presentation
    A 32-day-old girl with fever and restlessness was admitted in the hospital. Left facial swelling was found during physical examination. Redness was observed in the face. Prenatal history was normal. Birth weight was 3500 g. Body weight, length, and head circumference were 4300 g (75 th percentile), 52 cm (50 th percentile), and 38 cm (75 th percentile), respectively. She was breastfed. Pulse and respiratory rates were 130/min and 50/min, respectively. Axillary temperature was 37.8°C. Head examination revealed normal sized fontanel (1.5 × 1.5 cm) without bulging. Eye and ear were normal. Abdominal examination revealed no abnormal findings. Results of urine analysis and culture were normal. Blood urea nitrogen, sodium, potassium, and blood sugar were normal. Blood amylase was 10 U/L. Bilateral multiple reactive lymph node (size = 6 × 10 mm) at anterior cervical chain with a left facial swelling was observed in ultrasonography report. Pus was obtained following gentle pressure on Stensen’s duct. Staphylococcus aureus was detected in the microscopic and microbiological evaluations.The patient received a seven-day treatment course with vancomycin and amikacin. Neonate was discharged in a good condition..
    Conclusions
    Acute suppurative parotitis should be suspected in infants with fever, and irritability in pre-auricular region; and should be treated with appropriate antibiotics..
    Keywords: Parotitis, Staphylococcus aureus, Fever, Stensen's Duct
  • Seyed Mohammad Alavi *, Mohammad Mehdi Khoshkho, Shokrolah Salmanzadeh, Mehdi Eghtesad Page 11
    Background
    Diabetes mellitus (DM) due to suppressive effect on cellular immunity can impact on progression of tuberculosis (TB)..
    Objectives
    The aim of this study was to investigate the impact of DM on the epidemiological, clinical and para clinical aspects of pulmonary TB..Patients and
    Methods
    The information of 148 admitted pulmonary TB patients in infectious ward of Razi hospital in Ahvaz from 2009 to 2010 was extracted from their medical files. The patients were divided into two groups as TB with DM (n = 36) and TB without DM (n = 112). The related data on epidemiology, signs, symptoms, radiology and sputum smear examination in both groups were compared in SPSS 16 by using chi squared test..
    Results
    The mean age of TB with DM patients was higher TB without DM patients (56.6 ± 12.7 vs. 44.8 ± 18.3; respectively, P = 0.006). Whereas cough, night sweating, fever and weigh loss was not statistically different, sputum, hemoptysis and dyspnea was more prominent in TB with DM (69.4%, 33.4%, 44.5% vs. 36.6%, 9.8%, 20.5%; P = 0.005, P = 0.001, P = 0.005, respectively). In chest x-ray, cavitation and reticulonodular pattern was more frequent in TB with DM (55.5%, 22.2% vs. 31.2%, 8% - P = 0.008, P = 0.02, respectively). The rate of sputum smear positivity in TB with DM and TB without DM was 66.6% and 47.3%, respectively (P = 0.03)..
    Conclusions
    According to the results of this study, in approach to every DM cases suffering of respiratory symptoms such as productive cough, hemoptysis and dyspnea in association with cavitation or miliary mottling in chest x-ray, pulmonary TB should be considered at the top of the differential diagnosis list..
    Keywords: Pulmonary tuberculosis, Diabetes Mellitus, Epidemiology, Clinical Features, Radiology
  • Roya Rafiee, Fereshteh Eftekhar *, Seyyed Ahmad Tabatabaei, Dariush Minaee Tehrani Page 12
    Background
    Pseudomonas aeruginosa is one of the most common causes of nosocomial infections. Resistance of P. aeruginosa to β-lactam antibiotics may be the result of acquired resistance through mutation and over production of various antibiotic inactivating enzymes. This research aimed to determine the prevalence of extended-spectrum β-lactamases (ESBL) and metallo β-lactamase (MBL) production as well as the presence of their related genes among AmpC β-lactamase producing P. aeruginosa isolated from burns..
    Objectives
    The current study aimed to determine the prevalence of class A ESBL and MBL production in relation to the presence of their related genes among AmpC β-lactamase producing P. aeruginosa isolated from burns..
    Materials And Methods
    The antimicrobial susceptibility of 51 P. aeruginosa isolates from patients with burns was examined against 13 antibiotics by the disc diffusion method. Minimum inhibitory concentrations (MIC) for imipenem and ceftazidime were measured by the microdilution method. AmpC production was detected by AmpC disc and the modified three-dimensional extract tests. ESBL phenotype was confirmed by the double disc synergy test (DDST). Presence of β-lactamase genes was detected by specific primers and polymerase chain reaction (PCR)..
    Results
    All isolates were multidrug resistant. AmpC, ESBL and MBL production were observed in 35 (68.6%), 20 (39.2%) and 19 (37.3%) isolates, respectively. Overall, 43 isolates (84.3%) carried β-lactamase genes, out of which 31 (60.8%) harbored blaAmpC, 20 (39.2%) had blaTEM and 11 (21.6%) carried blaPER-1 genes. Among the AmpC producers, two isolates (6.5%) carried blaAmpC + blaESBL, 13 (41.9%) had blaAmpC + blaMBL and six (19.4%) produced the three enzymes..
    Conclusions
    A high prevalence of multiple β-lactamase production was observed among the AmpC producers (60%), of which the majority co-produced AmpC and MBL. The current study results showed correlation between β-lactamase production and the presence of antibiotic resistance genes in the isolates..
    Keywords: Pseudomonas aeruginosa, Beta, Lactamase, blaAMPC, blaESBL, blaMBL
  • Nasrin Noohi, Gholamhosein Ebrahimipour, Mahdi Rohani, Malihe Talebi *, Mohammad Reza Pourshafie Page 13
    Background
    Lactic acid bacteria, especially Lactobacillus spp., have been considered as excellent probiotic microorganisms, because of their activities in reducing the enteric diseases and maintaining healthy poultry..
    Objectives
    The current study aimed to evaluate the phenotypic characteristics and the probiotic potentials of Lactobacillus spp. isolated from poultry..
    Materials And Methods
    A total of 168 lactic acid bacteria (LAB) were isolated from healthy six and twenty-one-day old chickens and their feed samples. The isolated bacteria were identified by morphological, biochemical, and molecular tests including Polymerase Chain Reaction (PCR) and 16S rRNA gene sequencing. Biochemical fingerprinting with Phene Plate system (Ph-P) was done and the acid and bile resistant lactobacilli were subjected to the antibiotic susceptibility test..
    Results
    Amongst all of the examined LAB, 30.3% were resistant to bile and acid. Most of the isolated LAB (57.1%) belonged to the genus Lactobacillus with Lactobacillus brevis (78.1%) as the dominant species followed by L. reuteri (16.6%), L. plantarum (3%), and L. vaginalis (2%). The remaining isolates were identified as Pediococcus spp. (42.9%). The Ph-P cluster analysis of 75 L. brevis and 16 L. reuteri strains showed high phenotypic diversity. Whilst the results of Ph-P typing from L. reuteri strains showed low phenotypic variations especially among the strains sensitive to acid and bile salts..
    Conclusions
    Overall, the results showed that some of the high potential probiotic LAB species existed in Iranian poultry..
    Keywords: Lactobacillus, Poultry, Probiotic
  • Narges Cheraghchi, Pejvak Khaki, Soheila Moradi Bidhendi *, Azar Sabokbar Page 14
    Background
    Salmonella spp. is the major bacterial pathogen in poultry and is responsible for significant economic losses of the poultry industry in many parts of the world. Among Salmonella spp., Salmonella gallinarum and S. pullorum are the most common causative agents of chicken salmonellosis resulting in high mortality and morbidity..
    Objectives
    The aim of this study was to identify S. gallinarum and S. pullorum by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method..
    Materials And Methods
    In this study, 13 samples of Salmonella, isolated from local poultry, were obtained from Razi Type Culture Collection (RTCC). For the PCR-RFLP method based on the fliC gene, extracted DNA was used as a template for amplifying of the fliC gene (197bp) using specific primers. PCR products were subjected to digestion using Hinp1I restriction endonuclease..
    Results
    For the PCR, 197 bp fliC fragment was amplified from all 13 isolates. Ten out of 13 were S. gallinarum and the other three were S. pullorum. As part of the PCR-RFLP, two fragments were obtained (82 bp and 115 bp) for all S. gallinarum, whereas no digestion was observed in S. pullorum, and 197 bp fragment was seen..
    Conclusions
    PCR-RFLP with fliC gene and Hinp1I endonuclease were successfully applied to differentiate the two biotypes. The results suggested that this technique could be effective in detecting S. gallinarum and S. pullorum..
    Keywords: Salmonella gallinarum, Salmonella pullorum, RFLP, Hinp1I Endonuclease
  • Parisa Tajer Mohammad Ghazvini, Rouha Kasra Kermanshahi *, Ahmad Nozad Golikand, Majid Sadeghizadeh Page 15
    Background
    Magnetotactic bacteria (MTB) have the ability to biomineralize unique intracellular magnetic nanosize particles. These bacteria and their magnetosomes are under special attraction because of their great useful potential in nano-biotechnological and biomedical applications. MTB are ubiquitous in aquatic environments, but their isolation and axenic cultivation in pure culture is very difficult and only a limited number of them have been isolated in pure culture..
    Objectives
    The main goal of this study was screening, isolation and cultivation of a new strain of these fastidious bacteria in pure culture from Iran to use them and their magnetosomes..
    Materials And Methods
    Thirty samples were collected from various aquatic habitats. Most important physicochemical environmental factors that are involved in growth of MTB in the microcosms were investigated using inductively coupled plasma atomic emission spectroscopy (ICP-AES), portable dissolved oxygen meter, etc. Capillary racetrack technique and magnetic separation were used to purify and enrich MTB. Various isolation media were simultaneously used for isolation of a new magnetotactic bacterium in pure culture. Two imaging techniques were used to visualize the characterizations and cell division: transmission electron microscopy (TEM) and field-emission scanning electron microscopy (FESEM). Polymerase chain reaction (PCR), ChromasPro software and MEGA5 were applied for sequence analysis of the 16S rRNA gene..
    Results
    The results revealed a correlation of important physicochemical factors such as pH and iron with growth and blooms of these bacteria in the microcosms. New strain MTB-KTN90 was isolated in a modified isolation medium at microaerophilic zone from Anzali lagoon, Iran and cultured in a modified growth medium subsequently. The phylogenetic analysis showed that the strain belongs to Alphaproteobacteria. Growth and iron uptake studies indicated an important role by this bacterium in the iron biogeochemical cycle. For the first time, this paper introduced a cultured magnetotactic Alphaproteobacterium, able to synthesize magnetosomes in the temperatures above 30°C and reduce selenate oxyanion..
    Conclusions
    This paper may serve as a guide to screening, isolation, and cultivation of more new MTB. The new isolated strain opens up good opportunities for biotechnological applications such as medicine to bioremediation processes due to its unique abilities..
    Keywords: Magnetic Nanoparticles, Magnetotactic Bacteria, 16S rRNA Phylogeny, Alphaproteobacteria, Anzali Lagoon
  • Hamid Tebyanian, Seyed Hanif Mirhosseiny, Babak Kheirkhah *, Mehdi Hassanshahian, Hamze Farhadian Page 16
    Background
    Mycoplasma synoviae is an important avian pathogen which can cause both respiratory disease and synovial joint inflammation (synovitis) in poultry. Mycoplasmas spp. may cause the respiratory system infection in ostriches with symptoms such as inflammation of the nose, trachea and also damages of lungs..
    Objectives
    The current study aimed to use the M. synoviae specific Polymerase Chain Reaction (PCR) and microbiological methods in order to isolate and identify M. synoviae from suspected ostriches in Kerman Province, Iran, and compare the two methods (microbiological and PCR) employed to confirm Mycoplasmal contamination of ostrich lungs..
    Materials And Methods
    Fifty three samples of different parts of lung and trachea were immediately collected after slaughtering the ostriches in Kerman Province six months. Samples were cultured in the same conditions in pleuropneumonia-like organism (PPLO) broth and to isolate and identify M. synoviae, PCR and microbiological methods were conducted. The identified isolates were confirmed by specific amplification of 16S rRNA gene (163 and 207 base pair)..
    Results
    In the current study, 25 and 17 out of 53 ostrich samples were identified as Mycoplasma-positive in the PCR and microbiological methods, respectively; and 13 out of 25 the mentioned Mycoplasma-positive samples were also confirmed by PCR method..
    Conclusions
    The current study showed that PCR method is time consuming, effective, and efficient method to detect M. synoviae infection in ostriches. PCR method could be recommended as an alternative for culturing; M. synoviae was isolated from ostriches for first time in Kerman Province, Iran..
    Keywords: Mycoplasma synoviae, PCR, Microbiological methods
  • Babak Soltani *, Abbas Taghavi Ardakani, Alireza Moravveji, Mahzad Erami, Mostafa Haji Rezaei, Rezvan Moniri, Mansoor Namazi Page 17
    Background
    Nasal colonization of healthy children with Staphylococcus aureus is an important risk factor for different infections. Detection of colonized individuals with methicillin resistant S. aureus (MRSA) and its eradication is the proper prevention strategy for infection spread in the community and health-care centers..
    Objectives
    The aim of this study was to determine the prevalence, associated risk factors and antibiotic resistance pattern among healthy children who were nasal carriers of S. aureus..Patients and
    Methods
    This cross-sectional study was conducted on 350 one month to 14-year-old healthy children living in Kashan/Iran. The nasal specimens were cultured in blood agar medium for S. aureus. Positive cultures were evaluated for cephalothin, co-trimoxazole, clindamycin, ciprofloxacin, oxacillin and vancomycin susceptibility by the disc diffusion method and E-test. Risk factors for nasal carriage of S. aureus and MRSA were evaluated..
    Results
    Frequency of S. aureus nasal carriage was 92 from 350 cases (26.2%), amongst which 33 (35.9%) were MRSA. Isolates indicated an overall resistance of 52.2% to cephalothin, 33.7% to co-trimoxazol, 26.1% to ciprofloxacin, 26.1% to clindamycin, 35.9% to oxacillin and 4.3% to vancomycin. Factors associated with MRSA nasal carriage included gender (P value 0.001), age of less than four years (P value 0.016), number of individuals in the family (P value < 0.001), antibiotic use (P value < 0.001) and admission (P value < 0.001) during the previous three months, parental smoking (P value < 0.001) and sleeping with parents (P value 0.022)..
    Conclusions
    Age of less than four years, male sex, family size being more than four, antibiotic use and admission during the previous three months, parental smoking and sleeping with parents were independent risk factors for nasal colonization with MRSA..
    Keywords: Nasal Colonization, Staphylococcus aureus, MRSA, Risk Factors