[Antifungal activity of total extract and chloroform, ethyl acetate, methanol and aqueous fractions of aerial parts of Ephedra pachyclada against fungal strains]

Increased consumption of synthetic antifungal compounds has created serious side effects and wide spread anti fungal resistance. This persuades researchers to look for effective herbal plants as an alternative option. We report the results of anti fungal properties of different crude extract and relevant fraction of aerial parts of Ephedra pachyclada. In this experimental study, extracts and fractions of Ephedra pachyclada were obtained by maceration standard methods. The crude extract and fractions were diluted in defferent values from 31.25 to 500mg/ml. Candida albicans, Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, and Fusarium oxysporium were used for anti fungal activity assessment using standard agar diffusion methods. Total extract and chloroform, ethyl acetate, methanol and aqueous fractions had antifungal effects against Candida albicans only. We concluded that this plant contains antifungal compounds on Candida albicans, so this experiments and evaluation are pre-requirement of any applicable recommendation

[Evaluation of antibiotic resistance in bacteria isolated from patients hospitalized in imam khomeini and burn hospitals in Ahvaz City, Iran]

In recent years the emergence of antibiotic resistance has a high prevalence, so that it has become one of the complexities in modern medicine. This study was performed with the aim of evaluating the bacteria isolated from clinical samples of the patients with various infections and estimating the prevalence of various bacteria and also antibiotic resistance pattern. At first, culture was prepared from wounds of the patients with nosocomial infection in Imam Khomeini and Burn hospitals in 2008-2009. Then, after isolation of bacteria, antibiotic susceptibility was determined using antibiotic discs. Data were analyzed using statistical tests [ratio] at the significance level of p

Simultaneous detection and differentiates of Brucella abortus and Brucella melitensis by combinatorial PCR

OBJECTIVE@#To evaluate simultaneous detection and differentiates of Brucella abortus (B. abortus) and Brucella melitensis (B. melitensis) through the combinatorial PCR method.@*METHODS@#This study was designed using three primers that could simultaneously identify and differentiate two major species of pathogenic Brucella in humans and animals. Identification and differentiation of each species using the size of the PCR product were determined. To determine the specificity of the method, bacteria close to the genus Brucella were used. Finally, to confirm PCR products, In addition to the products sequence, RFLP was performed on PCR products using restriction enzymes.@*RESULTS@#The method of optimized combinatorial PCR in this study could simultaneously detect and differentiate B. abortus and B. melitensis with high specificity and sensitivity in clinical samples. Differentiation of species is based on the resulting bands; therefore, the band 494 bp for B. abortus and 733 bp for B. melitensis were obtained. RFLP and sequencing results confirmed PCR results.@*CONCLUSIONS@#The results of this study shows that without routine diagnostic methods such as culture and serology tests, using the molecular method of combinatorial PCR, important species of Brucella can be simultaneously identified and differentiated in clinical samples.

[Incidence of van A, B, C, D, E in vancomycin resistant enterococcus isolated from fecal flora in Tehran]

Vancomycin-resistant enterococci [VRE] have emerged worldwide and have become an increasing problem in clinical settings. Acquired glycopeptide resistance in Enterococcus species is due to the acquisition of van A, van B, van D, van C and van E genes, resulting in the production of peptidoglycan precursors with reduced affinity for glycopeptide antibiotics. The origin of these van genes is still unknown, but recent studies have indicated that van B resistance in enterococci might arise from gene transfer from the human bowel flora. In this study, we investigated the presence of Enterococcus-associated van A, van B, van C, van D, van E genes in the feces of hospitalized patients. To determine the prevalence of vancomycin-resistant enterococci [VRE] fecal colonization of hospitalized patients, 422 Enterococcus spp. isolated from stool of patients in Amiralam hospital. Disk diffusion method was used to detect resistance to vancomycin. The MICs of vancomycin were determined by the agar dilution method. The presence of van A, B, C, D and E genes were assassed by PCR analysis. PCR was positive for van A for 6 out of 10 [60%] and van B for 4 out of 10 [40%] of VRE strains. Among the van positive enterococci, two [20%] specimens contained both van A and van B gene, whereas no van C, D and E positive enterococcal isolates were identified from these specimens. The MIC of VRE isolates were between 512- 1024 micro g/ml. Our results showed that most glycopeptide resistant Enterococcus isolated from stool of hospitalized patients carried van A and van B. It is also possible that frequency of infections caused by glycopeptide-resistant enterococci will increase in our geographical area

Detection of Brucella abortus by alkB and IS711 based primers

Brucellosis is a zoonotic disease, which involves both animals and human. Although the conventional methods have been widely used for its laboratory diagnosis, the PCR techniques have proved to be useful due to specificity, sensitivity and the rapidness. Various target sequences of brucella bacterium such as OMP2, 16s RNA and IS711 have been used for the primer designing. All primer sets have shown different sensitivities and specificities. In present investigation, PCR protocol and primer designated based on IS711 and a fragment of chromosomal DNA all were optimized with standard genome and clinical samples. Numerous tissue samples [liver, kidney, lymph node, and uterus] were prepared and were cultured by the bacteriological standard methods along with the serology positive human samples. PCR protocol was optimized and the primer's sensitivity and the specificity were checked using pure genome of B. abortus. All samples were tested by the standard bacteriological methods. The samples were then subject to PCR amplification and the PCR product was confirmed using the RFLP technique. The culture results indicated a poor sensitivity as it was previously reported. The PCR product 157 bp was observed on the agarose gel indicating that significant number of clinical samples [human brucellosis cases] were positive by PCR but not by the culture method. Although B. abortus DNA was detected in all the culture positive veterinary specimens, some cross-reactions with close related bacteria were observed that might influence the interpretation of the results. The sensitivity of the present PCR protocol was significantly higher when alk B and IS711 based primers were used in compare to each of the alkB and IS711 based primers alone. More research will be needed to improve the specificity and sensitivity of the PCR protocol before recommending for routine laboratory works

Collagenase activity in prevotella bivius Isolated from patients with premature rupture of membranes

Bacterial vaginosis [BV] has been considered to be the most prevalent infection found in sexually active women. BV is thought to paly an important role in the premature rupture of membranes [PROM] and preterm birth. Preterm delivery accounts for a substantial percentage of low birth weight infants and perinatal mortality and morbidity. Bacteroides and Prevotella species have been isolated from the amniochorion of women with preterm birth and PROM. Women with >10 [4]/mL Prevotella bivius [formerly Bacteroides bivius] have a 60-100% higher rate of preterm delivery. The purpose of this study was to determine wether some strains of Prevotella species isolated from PROM and BV patients produce proteases especially collagenase enzymess which faciliate the rupture of membranes leading to preterm birth. Vaginal specimens have been obtained from 120 women with BV and premature rupture of membrane in 30-44 weeksss gestational age. Twenty anaerobic coccobacilli consisiting of Bacteroides fragilis, black pigmented Bacteroides and Prevotella bivius were isolated and identiofied. The isolates were examined for protease activity, using porcine skin gelatin and casein as substrates by Martleys method. Elastase and collagenase activity were detected using elastin, guinea pig collagen, bovine Achilles tendon collagen, FALGPA and GP, VPR as substrates by leverson method. Collagenase and elatase activity was detected in 90 and 100% of isolates. Gelatinase and caseinase activity was detected in 40 and 50% of isolates. Collagenase produced by Prevotella bivia isolates was purrified by ammonium sulphate precipitation, gel filtration and ion exchange chromatography. The collagenase was cleaved from the synthetic collagen substrate FALGPA, and GP, VPK substrates. The activity of the enzyme was inhibited by EDTA, Antipain and PMSF. This study suggests that proteases produced by Prevotella bivia may be involved in pathogenesis of premature rupture of membrane. PROM before 37 weeks has been reported to be significantly higher among patients with Bacteroides and Prevotella colonisation of the genital tract. The amniochorion consists of collage and elastin which convey physical integrity to the placenta. Collagenase and elastase released into genital tract may promote connective tissue destruction in the cervix and chorioammion membranes