Comparison of paraffin bait, humic acid vitamin b agar and paraffin agar methods to isolate nocardia from soil

The Isolation of Nocardia species is complex and time-consuming, which is due to rapid growth of adjacent bacteria. Because of the importance of a specific medium with the ability of controlling intrusive microorganisms, this study aimed at comparing three laboratory methods to introduce the reliable isolation technique for Nocardia species. The soil samples were collected from different regions of Tehran province, Iran, and carefully transferred to the laboratory. The samples were cultured in three different media including Paraffin Baiting,Humic acid vitamin B agar and Paraffin agar, and incubated for 3-4 weeks at 35 °C. Of 110 soil samples, 31 Nocardia isolates [28.18%] were obtained from the media including Paraffin Baiting, [19; 17.27%], Humic acid and vitamin B agar [4; 3.63%], and Paraffin agar,[8; 7.27%].because of high rate of isolation, low cost and the clearance of colonies suspected nocardia, Paraffin Bait technique is more reliable and efficient compared to the other methods

Detection of leptospires serogroups, which are common causes of human acute leptospirosis in Guilan, northern Iran

This study is performed to reveal most common species and subspecies of leptospires that are main causes of human leptospirosis in Guilan, Northern Province of Iran. We performed IgM-ELISA and MAT on 282 blood samples from patients who attended to 3 hospitals in the flat area of Guilan Province with clinical symptoms consisted with leptospirosis. All specimens with titers >/= 160 against at least one pathogenic strain in MAT and with titers >/= 160 in IgM-ELISA were regarded confirmed positive cases indicative acute disease. For any confirmed positive cases, we determined the strains, which had the highest titer to determine the frequency of most common serovars and serogroups. Seventy of 282 sera had titers >/= 160 against at least one pathogenic strain in MAT and titers >/= 160 in IgM-ELISA. We determined frequency of common causative serogroups which had highest titers in 70 positive cases and only cases which had high titers in MAT and in IgM-ELISA were selected which is a reliable criterion to detect acute disease and to determine causative serogroup. Nine serogroups including sejroe, grippotyphosa, mini, ictero haemorrhagiae, celledoni, autumnalis, cynopteri, pomona, and javanica were more responsible of acute leptospirosis in Guilan

[Study of antibacterial properties of adiantum capillus-veneris extract on eight species of gram positive and negative bacteria]

Adiantum capillus-veneris L. is a traditional medicinal plant which was used in the treatment of bronchitis and coughs, and also to prevent hair loss. Methanolic extract of this plant has demonstrated significant antimicrobial activities. In this study the antibacterial properties of Adiantum capillus-veneris L. extract on eight species of Gram positive and negative bacteria were evaluated. The herbal sample of Adiantum capillus-veneris was collected during the summer [June-July] from the north region of Iran called Condoluse and identified by herbarium laboratory in Faculty of Pharmacy, Tehran University of Medical Sciences, where a voucher specimen is deposited. The sample was pretreated and extracted with methanol 96% by percolation method and then concentrated and stored in a safe bottle until the experiments started. By using dilution method different dilutions of the extract have been prepared [10%, 5%, 2.5% and 1.25%]. Antimicrobial activity of the methnolic extract of Adiantum capillus-veneris were evaluated against 8 strain of Gram positive and Gram negative bacteria using agar disk diffusion and agar well plate methods. Our results demonstrated that prepared dilutions of the Adiantum capillus-veneris extract had significant effects on Staphylococcus aureus, Escherichia coli and Helicobacter pylori strains. Also the results indicated no significant inhibitory effects on Salmonella typhi, Shigella sonnei, Pseudomonas aeroginosa, Proteus vulgaris and Streptococcus pyogenes strains. Consistent with the other studies, our investigation demonstrated some antimicrobial effects of Adiantum capillus-veneris extract. With respect to various and multiple chemical properties of this plant, it is suggested that Adiantum capillus-veneris can be used for more medical and therapeutic purposes

[Evaluation of antibacterial properties and review of 10 medicinal herbs on preventing the growth of pathogenic nocardia species]

Historical documents belonging to medicine and pharmacy consist of valuable information about the herb and medicinal plants. The main purpose of this study was to investigate the antibacterial and phytochemical properties of 10 selected medicinal plants including of Echinophora platyloba D.C., Eucalyptus globules Labill, Euphorbia denticulate Lam., Hypericum perforatum L., Hypericum hyssopifolium Chaix, Hypericm scabrum L., Juniperus excelsa M.B., Peganum harmala L., Plantago major L. and Scirpus lacustris L. in preventing the growth of pathogenic Nocardia in-vitro. All species of Nocardia are found in nature in the soil and aquatic environments. The bacteria are aerobic, gram-positive, partially acid fast bacilli. Nocardia asteroids a highly pathogenic microorganism infects humans through the respiratory tract. The bacterium is primarily an opportunistic pathogen that causes the infection in patients with immunodeficiency. Nocardia brasiliensis is most commonly responsible for cutaneous infections. The antibacterial ability and phytochemical properties of methanolic extract of the above herbs against species of pathogenic Nocardia were investigated by means of appropriate agar distribution methods. After incubating of the cultured media, the diameter of the clean zone formed in each concentration was measured and correlated to the ability of the extracts to inhibit the growth of Nocarida strains. According to the results, the various concentrations of the 5 medicinal plants out of the 10 herbs e.g. Hypericum perforatum L., Hypericum, hyssopifolium Chaix, Hypericum scabrum L., Juniperus excelsa M.B., Scirpus lacustris L. have significant antibacterial activity against Nocardia asteroids and N. brasiliensis. Interestingly the results show inhibitory effects of the medicinal plants on different strains of the Nocarida in-vitro. It was found that the 5 medicinal plants out of the 10 herbs have potent antibacterial property. We came to this conclusion formed by 10% concentration of the above herbal extracts in appropriate distribution routs, were similar to that of the clear zones formed by specific antibiotic against Nocardia species, e.g. Amikacin, Amioxicillin, Ceftazidime, Ceftizoxime/Cefotaxime, Cephalothin, Cotrimoxazol. The antibacterial activity of the above herbs is probably associated to the potency of strong antibacterial properties of flavonoids, saponins and tanans presents in the methanolic extraction of the above medicinal plants

Evaluation of indirect fluorescent antibody assay for detection of Nocardia

Nocardiosis is an acute or suppurative chronic disease caused by an aerobic, gram-positive, weakly acid-fast and soil-borne filamentous and organism. Nocardia asteroides which is the dangerous and most frequently pathogen, infects humans through the respiratory tract. The bacterium is primarily an opportunistic pathogen that causes the infection in patients with underlying immunodeficiencies. The main purpose of the study was to detect antibody titre against Nocardia in all study groups, using indirect immunofluorescent assay [IFA]. Correlation between the antibody titre against Nocardia with age, sex, occupation, and chronic pulmonary infection and corticosteroid therapy patients was also investigated. The present investigation is a Cross-Sectional study conducted on a population consisted of 300 subjects including 200 hospitalized individuals patients, nurses and healthcare workers from Imam Khomeini hospital, and 100 health adult blood donors. None of the patients had already been diagnosed to be affected by Nocardia. Our results demonstrated four patients suffering from different infections, including TB, mycetoma, chronic pulmonary and chronic obstructive pulmonary diseases were IFA positive. None of the high risk hospital personnel, who were working in close proximity to the areas infected with Nocardia, were found to be IFA positive. Meanwhile there was no positive result in a group of patients [n=34] who were under corticosteroid therapy. Finally, considering the small sample size of the IFA positive cases no significant association between the IFA results and age, sex, occupation and clinical conditions of the subjects could be established

[ prevalence and molecular characterization of vancomycin resistant gram positive cocci isolated from patients in Tehran]

Gram positive bacteria, particularly, Staphylococcus aureus, coagulase negative staphylococci and enterococci are of particular concern in hospitals. But there has been increasing concern about the development of vancomycin resistant enterococci and MRSA strains with reduced susceptibility to vancomycin over the last decade. Therefore, the present study was carried out to confirm the identification of vancomycin resistant gram positive cocci, to determine antibiotic resistance pattern and to study vancomycin resistance genes. The isolates from clinical samples were collected from hospitalized patients and outpatients in Tehran. Gram positive cocci species identification was performed by using conventional tests and PCR using specific primers. VRE isolates were subjected to antibiotic susceptibility testing, MICs of vancomycin were determined by the E-test method. Determination of vancomycin resistance genes, vanA and vanB were performed with PCR. Confirmation of transposons was performed with specific primers for vanS. Out of 1030 gram positive isolates, none of the staphylococci or streptococci isolates were resistant to vancomycin. Most of vancomycin resistant isolates in this study were VRE. faecium [96%] and harbored vanA. All of the isolates were positive for vanS the conserved fragment of transposon and carried the identical digestion pattern like type strain. According to the results of this study, all of the vancomycin resistant isolates were enterococcus spp. Vancomycin resistant enterococci itself is now a major and largely untreatable infection, and can pass the vancomycin resistance genes to the other highly virulent gram positive cocci

Distribution of human leptospirosis in guilan province, Northern Iran

Leptospirosis is the most widespread zoonosis in the world and is more prevalent in tropical and temperate regions. Guilan Province in north of Iran, is an endemic region of human leptospirosis. Since diagnosis of leptospirosis according to clinical symptoms is very difficult due to lack of characteristic pathogonomic sign[s], laboratory support is necessary. In 2003, we obtained blood samples from patients hospitalized in main general hospitals of Guilan Province and were suspected as having leptospirosis according to their clinical presentations. We examined 995 sera by a commercial IgM and IgG ELISA kit to find positive cases. 62.7% of positive cases were male and about 86% of them were farmer. High distribution rate of leptospirosis was seen in middle-aged people [65% in 20-50 years old]. It seems that leptospirosis has a high occurrence in major cities and is mostly distributed in warm months of the year. Demographic analysis of the results indicates that leptospirosis is typically a rural and an occupational disease in the area

Characterization of vancomycin resistant enterococcus faecium

To determine the species distribution, updated drug susceptibility patterns and genes conferring resistance in clinical vancomycin resistant enterococcal [VRE] isolates. Clinical enterococcal isolates collected during 7 months, from September 2005 to April 2006 from hospitalized patients and outpatients were studied. Twenty five VRE were isolated from 450 enterococci samples [5.6%]. VRE isolates were subjected to antibiotic susceptibility tests. Genotype of these isolates was determined by PCR. All of the isolates were E. faecium and carried the vanA gene. Antibiotic susceptibility tests showed that the isolates were resistant to ampicillin 25[100%], ciprofloxacin 25[100%], gentamicin 24[96%], erythromycin 25[100%], tetracyclin 10[40%] and chloramphenicol 2[8%]. VRE strains were resistant to three antibiotics and were susceptible to new antibiotics linezolid and dalfopristin- quinupristin. Switching to treatment with these antibiotics would relieve the problem for a short time

[Isolation and characterization of streptomyces in composted Iranian soil, using chemotaxonomy]

The Actinomycetes are gram-positive organisms that tend to grow slowly as branching filaments. They were thought to be related to both bacteria and fungi, but in recent years it has been undoubtedly shown that they are prokaryotic organisms. Actinomycetes have proved to be causal agents of many human and animal infections. They also play as an important rule in producing new antibiotic agents. Fifty soil samples collected from different regions of the country were analyzed to determine the presence and types of antibiotic-producing Streptomycetes using dilution plating method. Starch casein agar and glucose yeast extract agar were used as the culture media. 88 selected organisms from 140 isolates were subjected to chemotaxonomy determination. The isomeric form of diaminopimelic acid [DAP] was determined by thin-layer chromatography [TLC] of hydrolyzed whole-organism preparations. A sample of the hydrolysated extract was applied as a 2[cm] band from the base of the TLC plates. A standard containing meso and LL-DAP was also run at both sides of the sheet for comparison with test mixtures. The plates were developed for about 3 [1/2] hours in a glass tank containing 40 ml methanol, 2 ml 6Mol HCl, 5ml Pyridine, and 13 ml water. DAP appears as green/brown spots, changing to yellowish with time. The LL-DAP, meso-DAP and hydroxy-DAP isomers have approximate R[f] values. Approximately 97% of test strains contained LL-DAP and the remaining 3%, contained meso-DAP. This study demonstrated a wide variety of Streptomycetes bacteria isolates which are capable to produce antibiotics and TLC is an appropriate method in determination of Them

[Evaluation of two commercial qualitative and semi quantitative ELISA methods for diagnosis of acute human leptospirosis]

Introduction: Leptospirosis is most widespread Zoonosis in the world, especially in Tropical and temperate regions with moist climate. Diagnosis of Leptospirosis according to clinical symptoms is difficult and uncertain due to lack of specific sign [s]. Leptospira is a fastidious bacterium. Isolation of these bacteria by culture is difficult, time consuming and hence doses not contribute to an early diagnosis. Specific antibodies against Leptospira appear from 6th day of disease onset. The Microscopic Agglutination Test [MAT] is most reliable assay but generally requires paired sera for detection of seroconversion and is considered too complex for routine use

Objective: We performed this study to final a simple and reliable method for diagnosis of acute human leptospirosis. ELISA is another common method for diagnosis of Leptospirosis

Materials and Methods: In summer of 2004, we examined 282 single sera of patients who were suspected of Leptospirosis by a commercial none quantities and an in-house semi quantative ELISA assays and compared their results with MAT

Results: Mean time of first taking blood samples were 6:36 after onset of symptoms. All specimens with titers >/=640 against a pathogenic serovar in MAT were regarded as confirmed leptospirosis [104 from 282]. All specimens which were positive in any IgM-ELISA assays were compared with the results of MAT. In our study, sensitivity, specifity, positive and negative predictive values of in house ELISA were 89.4%, 87%, 80.2% and 93.4% consequently but were 87.5%, 41.5%, 44.6% and 85% for comerical IgM-ELISA assay consequently

Conclusion: The results of our study show that IgM- ELISA assay is a reliable and sensitive method for diagnosis of acute leptospirosis and also show that in house semi quantative IgM ELISA was more specific and commercial qualitative IgM- ELISA was more sensitive

diagnostic study of nocardiosis patients being confined in Shareati training hospital in Tehran, using cultural and serological methods

The present investigation was carried out to detect nocardiosis in immunocompromised patients confined in the pulmonary ward of Tehran's Shareati Training Hospital through the use of indirect immunofluorescence assay [IFA] and bacterial culture methods. The comparison of the two methods and the correlation between the antibody titer and the statistical and epidemiological data were also investigated. One hundred and one patients with advanced symptomatic pulmonary infection were studied in the course of a twenty-month period. Individual patients' sputum, BAL [bronchoalveolar lavage] and blood sera were tested. From each sample three thin smears were prepared for microscopic observations. The samples were cultured in Sabouraud's dextrose, blood and paraffin agar. The detection of antibody against Nocardia asteroides was carried out in all study groups, using the IFA method. The medical history of patients was also obtained through questionnaires for further analysis. Forty one patients suspected for Nocardiosis with an antibody titer ranging from 1/4 to 1/512, detected by IFA method, included 26 [63.4%] men and 15 [14.8%] women. The age of the patients varied from 7- 80 years. Those with reasonable antibody titers included 15 [36.5%] housewives and 9 [21.9%] workers. Nocardia asteroides was isolated from only one patient suffering from Wagner vasculitis with an antibody titer of 1/512 in serum. Furthermore, in-vitro investigation for the differentiation of the isolates was performed and confirmed the notion that the organism which grew on the primary media was, indeed, the Nocardia asteroids complex. Our results revealed that the bronchopulmonary infections, which occur in high-risk patients T-cell deficiencies, long term corticosteroid therapy, immunocompromised hosts, HIV infection, organ transplantation- was an important index for the primary diagnosis of Nocardiosis. As the important finding of the present research, antibody titer of 1/64 could be taken as a cut-off value for diagnosis of patients infected with Nocardia, though lower titers should not be totally ignored

epidemiology of ieptospirosis in guilan provice 2003

Leptospirosis is most widespread Zoonosis in the world and is more prevalent in tropical and temperate regions. In most Of Iran, climatologic and ecological conditions are unfavorable for leptospirosis to play an important role as a public health problem. However this does not count for the neglected flat area of Guilan province which presumably represents a region with a high incidence of human leptospirosis. This area has a subtropical climate with mainly farmers as inhabitants. By far the most important agricultural activity is rice farming. Most of farmers used to keep domestic animals in their houses and rodents are abundant. To find evidence for a high incidence of leptospirosis in the Guilan province, we collected blood samples from patients who attended one of the three big general hospitals in the province with clinical symptoms consistent with leptospirosis in 2003. All sera were stored at -20°C until examination by ELISA and MAT. All patients whose serum had titers

evaluation of the potent inhibitory effects of royal jelly fractions against streptomyces bacteria

To investigate the anti-bacterial properties of royal jelly against six different bacteria [Escherichia coil, Staphylococcus aureus, Streptomyces griseus, and three unclassified strains of Streptomyces]. Design: To find out the antibacterial properties of royal jelly by means of agar distribution method [drop-plate]. Setting: The diameter of the clean zone formed in each concentration was measured and correlated to the ability of the extracts to inhibit the growth of microorganisms. Interestingly the results show inhibitory effects of royal jelly on different strains of the bacteria in-vitro. Four concentrations of water soluble extracts of pure royal jelly were prepared and added drop wise to the bacterial strains seed layer cultured individually. Main outcome measures: Ether-soluble and non-soluble fractions of the royal jelly were also examined using the above method. It was found that the antibacterial effect of ether-soluble fraction is substantially higher than the effect of ether non-soluble fraction. The zones formed by ether-soluble fractions of royal jelly in the route of distribution in agar belong to the potency of anti-bacterial properties of fatty acids present in this fraction of royal jelly [10 -HDA]. Conclusions: The non ether-soluble fraction of royal jelly contains a bactericidal substance called royalisin that was found to have potent antibacterial activity. This fraction of royal jelly revealed weaker antibacterial effect than the ether-soluble fraction and even of pure royal jelly. Hence apart from its nutritional and medicinal effects, royal jelly's bactericidal ability should be considered and employed whenever appropriate in clinical medicine

Pulmonary Nocardiosis associated with Cushing's syndrome

Nocardia species consists of strict aerobic bacilli that form branched hyphae in both tissues and culture. Infection with Nocardia is an uncommon but important cause of morbidity and mortality in immunocompromised patients and organ transplant recipients. Since Nocardia species have been isolated from the soil of different regions of Iran, as well as different cases of Nocardiosis, this study was undertaken to evaluate the role and frequency of Nocardia in pulmonary infections in the country. The present study was planned to isolate Nocardia bacteria from immunocompromised patients who had been referred to Maseeh-e-Daneshvari Training Hospital [Tehran]. 142 patients with advanced symptomatic pulmonary disease were studied during a period of seven months. Of at[the patients surveyed 102 were tested both for their BAL [bronchoalveolar lavage] and sputum. For the rest of the patient's sputum samples were not accessible. From each sample three thin smears were prepared for microscopic observations. The samples were cultured in Sabouraud dextrose agar and paraffin agar. Medical histories of patients were also recorded in the certain questionnaire for further data analysis. Nocardia asteroids was isolated from only a patient suffering from Cushing's syndrome with bronchogenic carcinoma [Ectopic adrenocorticotrophic hormone [ACTH] syndrome]. Further in-vitro investigation for differentiation the isolate was performed and confirmed that the organism grew on primary media was Nocardia steroids complex. Results revealed that the normal concentration of NaOH [4%], which is routinely being used for identification of Mycobacteria species, could inhibit the growth of Nocardia. Therefore, decontamination procedure of the samples collected for isolation of Nocardia was performed using 1% NaOH in this study

Study of tuberculous infection rate in townships in a Centeral Province of Iran

Tuberculosis is a continuous threat for health in all parts of the world. An estimated one-third of the world' s population is infected with the Mycobacterium tuberculosis and 7 to 8 million people develop TB disease each year. The purpose of this study was to investigate the rate of tuberculosis in the townships of Yazd province, Iran. During the study period [19971999], 3885 suspected tuberculosis patients [1820 males and 2065 females; aged 8-85 years] who had been referred to the Yazd referral polyclinic were investigated by Ziehl Neelsen staining and culture method and questionnaire was completed for each subject. Then, Collected data were analyzed by statistical package for social science [SPSS] and chi-square program. The results show that, of the total suspected tuberculosis, 604 cases were found to be positive for tuberculosis. The average annual rate of tuberculosis was 26.8 cases per 100000 population [23.1/100000 males and 31/100000 females]. The highest and lowest rates of tuberculosis were observed among Sadough [78.1/100000] and Abarkouh townships population [19.8/100000] and also among age group >/= 50 years old [111/100000] and < 10 years old [7/100000], respectively. The average annual rates of pulmonary and extra-pulmonary tuberculosis in Yazd province were 152 cases [20.2%] and 48 cases [6.4%], respectively. It seems that, despite the efforts, which have been done for prevention, diagnosis and treatment of patients with tuberculosis, it is still considered as a threat for health in the Yazd province, Iran