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Abstract Bachground and objectives:Streptomyces is the most important genus in Actinomycetes family.The Streptomycetes are widely used in industry producing numerous chemical compounds including antibiotics, enzymes and anti- tumor agents. The aim of this study was to isolate soil-borne Streptomyces producing antimicrobial substances from soil of Golestan province of Iran and to survey anti-fungal metabolities produced by this organism. Material and Methods:In this study various soil samples were collected: ( forest areas of Naharkhoran in Gorgan and Kordkuy’s Derazno, Aghala’s deserts and agriculture lands of Aliabad ) were cultured on Actinomycet isolation agar and Starch casein agar were identified and purified by morphology and biochemistry tests. The activity of isolated Streptomyses against:Aspergillus flavus, Aspergillus, Candida albicans and Malasesia fur fur were studied by Agar Diffusion. Results:Of 120 samples, 24 are Streptomyces(20%) .The frequency of Streptomyces are reported in Aghala (10,41.6%),Derzno (8,33.3%) ,Nahar khoran(4,16.6%) and Aliabad(2,8.3%).Of 24 isolated Sterptomyses,two isolates have strong anti-fungal and six of them have moderate effect.We also see Streptomyses,isolated from desert area, have higher anti-fungal activity. Conclusion:It is recommended two isolated of Streptomyses be identified ana purified. Key words:Streptomyces , antifungal activities, antibiotic

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Abstract Background and objectives: Direct smear microscopy, because of its simplicity, rapidity, low cost and relatively sensitive is a suitable method to detecting pulmonary tuberculosis. This experiment was aimed at determining the best method for detecting Mycobacterium tuberculosis among three kinds of staining methods: Fluorochrome, Ziehl Neelsen and Tan Thiam Hok . Material and Methods: The sputum specimens (N=714) were obtained from people with suspected pulmonary tuberculosis and identified by three staining procedures. We determined the sensitivity, specificity, and the positive and negative predictive value of them and compared results by growth on Lowenstein-Jensen medium as gold standard. Results: Ninety-three (13%) of 714 sputum specimens were positive in culture method. The sensitivity of Tan Thiam Hok, Ziehl Neelsen and Fluorochrome are 89.2%, 91.3% and 95.6%, respectively, while their specificity and positive predictive value were 100%. Their negative predictive values were 98.3%, 98.7 %, 99.3 %, respectively. Conclusion: We conclude that Ziehl Neelsen can still be a reliable procedure for detecting AFB in sputum specimens because it has the appropriate sensitivity and specificity in comparison with another method of staining. Key words: pulmonary tuberculosis, Tan Thiam Hok staining, Ziehl-neelsen staining , Flurochrome staining

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Abstract Background and objectives: Genital tract infections are among the most common causes of patients referred to therapeutic centers. Nearly 75% of women suffer from genital Candida infection, at least once in their lifetime. The aim of present study was detection of Candida species causing vaginitis and the evaluation of antimycotic effects of ketoconazol, clotrimazole and fluconazole against Candida species. Material and Methods: In this study, 210 vaginal samples were obtained from the patients suspected of Vaginal Candidiasis. Direct examination and culture were carried out for all specimens to detect the yeast. The isolated yeast species were then identified, using various different tests such as culture on corn meal agar, tween-80, germ tube test, and assimilation test by API 20C kit by using Sabouraud Dextrose Agar and microdilution broth, MIC90 and MIC50 of drug were measured and determined their drug resistance. Results: In the present study, 100 yeast colonies were isolated from patients %80 are C. albicans and the rest are C. parapsilosis(2%), C. tropicalis(6%), C. glabrata(4%), C. krusei(2%), C. guilliermondii (3%), C.stellatoidea(3%). In terms of drug resistance test MIC50 and MIC90 of fluconazole for candida albicans are 5.33 and 35.27μg/ ml, respectively, and for non-albicans candida are 3 and 21.4μg/ml, respectively. Clotrimazole MIC for Candida albicans (MIC50, MIC90) 0.97 and 4.9μg/ml, respectively, and for non-albicans 0.63 and 3.4/ml, respectively. Kectoconazole MIC for Candida albicans 2.43 and 16.45μg/ml, respectively, and for non-albicans 1.12 and 6.6μg/ml, respectively. Conclusion: Clotrimazole has been better than the two other drugs for Candida species on the whole, non albicans species are more sensitive than albicans species in the presence of the drugs used in this study. Key words: Candida, vaginal candidiasis, Resistance drug , Tonekabon.

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Abstract Background and objectives: Giardiasis is a parasitic infection of small intestine, with a worldwide distribution and the prevalence of Giardia in different parts of the world varies between 1 to 25%. Plants have the vast range of antimicrobial and antifungal activity that can be identified as alternative treatments for bacterial and parasitic pathogens, the same as Giardia. In this study, the methanol extracts of eucalyptus plants, Satureia hortensis and Heracleum glabrescens, on Giardia cysts were studied in vitro. Material and Methods: The cysts were isolated from the feces using a modified Bingham. After counting by Hemusytumetr, they were placed near by 200 mg / ml, 100 mg / ml and 10 mg / ml of the extracts prepared by DMSO for 30 and 60 minutes. Then, the number of dead and live cysts was counted under a microscope. Results: the fatality effect of the extracts in 60 minutes is higher than those of 30 minutes. The methanol extracts of Satureia hortensis, Eucalyptus and Heracleum glabrescens with the dilution of 200 mg/ml in 60 mins have the fatality effect of 84/3%, 63/3% and 44%, respectively. The highest fatality(84.3%) on Giardia cysts is related to Satureia hortensis with the dilution of 200 mg/ml in 60 mins and the Lowest(27%) is related to Heracleum glabrescens with the dilution of 10 mg/ml in 30-minute period. The significant relationship between the plant type and the fatality of methanol extracts is observed. Conclusion: the methanol extracts of Eucalyptus, Heracleum glabrescens and especially Satureia hortensis have anti-parasitic effects in the laboratory conditions. Thus, they can be used in the future, instead of the chemical antiparasitic drugs. Key words: Antibacterial Giardia lamblia cysts, Eucalyptus, Satureia hortensis, Heracleum glabrescens, Tonekabon

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Abstract Background and Objective:Staphylococcusaureus is one of the important factors causing nosocomial infections. Typically 25-30 percent of healthy people carry the bacteria in their anterior nasal cavity. The physicians(50%), nurses (70%) and hospital staff (90%) are the carriers of this bacteria, leading to the infection of inpatients. The emergence of antibiotic-resistant Staphylococcus strains to vancomycin and methicillin has brought about several problems in treatment of the infections caused by Staphylococcus strains. Hence, we aimed to study the frequency of staphylococcus aureus carriers and resistance pattern among medical personnel of the surgical ward in ShahidRajaee hospital, Tonekabon. Material and Methods: this analytic-descriptive study was conducted on the samples taken from nasal carriage of medical staff of surgical ward (N=120). Antibiotic- resistant of Staphylococcus strains was assessed by antibiogram and disk diffusion (DAD), in accordance with CLSI standards. Results: of 34 (28.33%) who are nasal carriers of staphylococcus, 12 are over 30 years old and 24 under 30. Based on antibiogram, 1.97% of specimens are sensitive to Gentamicin and Co-trimoxazole, 1.94% to Ciprofloxacin, 2.88% to Vancomycin and 6.20% to Methicillin. In addition, 100% of specimens are resistant to Ampicillin, 1.97% to Penicillin and 2.88% to Amoxicillin. Four isolates areresistant,both to methicillin and vancomycin. Conclusion:In this study, the spectrum of S. aureus resistant and sensitive strains to some antibiotics is similar to other studies, but a dramatic increase is seen in the rate of MRSA and non-susceptible cases to vancomycin. The Effectiveness of Penicillin, Amoxicillin and Ampicillin is still very low on S. aureus samples. Key words: Prevalance Resistance Pattern, Staphylococcus aureus, Medical Staff, Nasal Cavity, Tonekabon

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Abstract Background and Objective: One of the most common diseases of keratin tissues is dermatophytosis caused by dermatophyte fungi. Because of being contagious, it has a high prevalence rate in wrestling and body building gyms. This study was designed to evaluate the process of this disease and improve the hygiene of halls. Material and Methods: The Samples (N= 540) were obtained from athletes and gyms, and a questionnaire was used to gather information. To identify various specious of dermatophyte, the routine diagnostic procedures, culture media, and supplementary tests were performed. Results: Of samples taken from athletes, 59 wrestlers and 11 body builders suffer from dermatophytosis. Trichophytontonsurans (%28.81) and Epidermophytonfloccosum (%36.36) are the main isolates in wrestlers and body builders. Also the rate of epidermophytonfloccosum (%37.5) is the highest in the samples taken from gym mats and halls. Conclusion: Because of high prevalence of dermatophytosis, pay attention to increase of hygiene and training courses for coaches and athletes are crucially important. Keywords: Dermatophytosis Wrestling and Body Building Halls Challous

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Abstract Background and objective: There is increasing evidence for the role of oxidative stress in female reproductive tract. The purpose of this study was to determine the activity of antioxidant enzymes during menstrual cycle. In addition, the relationship between activity of antioxidant enzyme and sex hormones was evaluated. Materials and methods: In this study the activity of superoxide dismutase, glutathione peroxidase, glutathione reductase, catalase and total antioxidant capacity during the menses, follicular and luteal phases of the menstrual cycle in twenty women with regular menstrual cycle were studied. Furthermore, the correlation between activity of antioxidant enzymes and estradiol, progesterone, LH, FSH and testosterone were evaluated. Results: There was no significant difference between activity of superoxide dismutase, glutathione peroxidase, glutathione reductase, catalase and total antioxidant capacity during the menses, follicular and luteal phases of the menstrual cycle (P>0.05). We found significant correlation, in luteal phase, between superoxide dismutase and FSH (P<0.05، r=0.44) and LH P<0.05،r=0.54). Also it is observed between LH and glutathione peroxidase (P<0.05، r=0.44). Conclusion: Based on the results, there is no significant difference between antioxidant enzymes and total antioxidant capacity of plasma during menstrual cycle. In other words, physiologic system of women with regular menstrual cycle can protect body against oxidative stress and this is probably performed due to action of FSH and LH hormones. Keywords: Antioxidants Menstrual cycle Sex hormones

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Abstract Background and Objective: Recognizing and using of isolated phytase in the soil microorganisms are paramount importance to produce the Phytase enzyme utilized commercially in different industries. This study was conducted to recognize different bacillus species which are Phytase producers and detection of the gene that can produce this enzyme. Material and Methods: Soil samples were gathered through different parts of mountainous areas. The early isolation of bacillus was carried out in Bacillus Medium Agar. After isolating the bacteria and genome extraction, the responsible gene of enzyme producer recognized and amplified by PCR method. The size of this protein and the optimal production situation in supplemental exploitation such as SDS-PAGE and the enzymatic activity of its size were evaluated. Results: Of 40 samples, one bacterium secreting Phytase enzyme was isolated. This bacterium was sequenced and recognized Bacillus Sobtlis species that is classified in STR Genus. The size of protein phytase produced by this gene was about 45 KD and the enzyme activity at 55 degrees was measured about 5.65 in wavelength of 415 NM. The phytase gene with the size of 1200 bp was propagated. Conclusion: the microorganisms, in natural conditions, produce Phytase enzyme in limited amount and with the quality appropriate to microorganisms. Thus, isolating the bacilli producing Phytase enzyme and purifying this protein are highly significant. Key words: Bacillus Subtilis Phytase SDS-PAGE Enzymatic Activity Polymerization Chain Reaction

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Abstract Background and Objective: Chitin, which is a linear polymer of N-acetyl glucosamine residues, has been the most abundant polymer in nature after cellulose. In recent decades, Chitinases have received increased attention because of their wide range of applications, especially in biological control against fungi. Material and Methods: the isolation of bacilli producing chitinolytic enzymes was performed by collecting 40 soil samples from various regions of Gorgan, northern of Iran. The chitinolytic potential of the isolates was indicated by observation of clear zone in colloidal chitin agar medium. Identification of selected strains was performed by polyphasic taxonomy, and subtler identification and sequensing were carried out by extraction DNA. Antifungal effect was evaluated by well method against Candida albicans (ATCC 10231) Aspergillusniger (ATCC 2029)،Aspergillusflavus (IR6) Fusariumoxyporum (PTCC 5115) and Alternariaalternata (PTCC 5224). Results: Nine colonies of chitinase positive bacillus were isolated on choloidal Chitin Agar (CCA) and five of them had antifungal effect. R6 strain had the highest, and R2 and R3 had the lowest effect on fungi. The 16S rRNA sequence of these isolations in comparison with the known bacteria has 95-97% similarity. Conclusion: Some of the soil bacteria can have antagonestic effects on human and phytopathogenic agents existed in soil. Keywords: Bacillus Chitinase Soil Antifungal

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Abstract Background and Objective: Enterococci are normal flora of human body and considered as the third leading cause of nosocomial infections. The aim of this study was to determine drug resistance of Enterococcus species through biochemical methods. Material and Methods: One hundred twenty-eight of enterococcus suspected samples were isolated from gorgan and gonbad’s hospitals from April to June, 2013. The samples were cultured on blood agar, chrome-agar, EMB agar and some special cultures of isolation of Enterococcus species. Suspension of bacteria was grown in Mueller Hinton agar and the inhibition zone diameter was determined by disk antibiogram. Results: Of 128 samples, 109(85.15%) were enterococci faecalis and 19 (14.85%) Enterococcus Faecium. In all of 128 cases, eight showed resistance to amoxicillin, ten to ampicillin, five to gentamicin, five to ciprofloxacin, six to chloramphenicol, four to cephalexin and one to vancomycin. Conclusion: It seems to be necessary to use drug sensitivity test for having appropriate treatment and preventing from resistance strains. Keywords: Enterococci, Antibiotic Resistance, Antibiogram

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Abstract Background and Objective: With the development of drug resistance in strains of fungi, there is a considerable resistance of Candida albicans strains to fluconazole. Molecular studies are developing to determine the relationship of such a drug resistance with the increased gene expression of enzymes produced in drug-resistant Candida isolates. We aimed to evaluate the relationship between extracellular lipase gene (LIP8) expression of Candida albicans isolated from candidiasis and sensitivity or resistance to fluconazole. Material and Methods: Drug susceptibility of Candida albicans was performed in oral and vaginal candidiasis to determine the proportion of strains sensitive or resistant to fluconazole using NCCLS method. To evaluate and compare the expression of these genes in the susceptible and resistant strains, RT real-time PCR reaction was used. Results: Of 46 Candida albicans, 20 were susceptible, 12 were semi-susceptible and 14 were resistant to fluconazole. By using PCR reaction, the results showed that the expression of this gene in fluconazole-susceptible isolates was moderate, while it was high in the isolates resistant to fluconazole. Conclusion: The results of lipase gene (LIP8) expression showed that the additional expression of some genes of the enzymes responsible for virulence of Candida may also play a role in resistance to fluconazole. Keywords: Candidiasis, Lipase Gene Expression, RT real-time PCR, Fluconazole

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Background and objective: Implementation of standard methods for accurate detection of bacteria, correct antibiotic susceptibility testing and effective treatment of bacterial infections play important roles in development of public health and prevention of drug resistance. This study aimed to detect bacteria using standard methods and compare the results with the results obtained in teaching hospitals’ laboratories.

Methods: Positive culture plates containing bacteria isolated from patients in hospital laboratories in city of Sari were transferred to microbiology laboratory of Faculty of Medicine at Mazandaran University of Medical Sciences, after determining the genus and species of bacteria and antibiotic susceptibility testing of the isolates. The samples were re-examined based on standard protocols, and antibiotic susceptibility testing was done using the Kirby-Bauer method.

Results: Of 101 patients, 20% of bacteria and 22.5% of antibiotic sensitivity results reported by the hospital laboratories were incorrect. There were significant differences between the two study groups in terms of bacterial species detection and sensitivity to some drugs (P<0.05).

Conclusion: In the present study, lack of implementation of internal quality control programs in some hospital laboratories and lack of proper monitoring by regulatory authorities in different departments of the hospital have caused 20% false-detection results in hospital reports. Inconsistency in results of laboratories, false antibiograms and subsequent false laboratory reports cause drug resistance in some patients. This indicates the necessity of continuous training in the field of Microbiology and implementation of standard protocols and methods for detection of bacterial species and antibiotic susceptibility testing.