Detection of Pseudomonas aeruginosa in sputum samples by modified fluorescent in situ hybridization

Pseudomonas aeruginosa is the most common and dominant infectious agent that causes chronic pneumonia in patients with cystic fibrosis (CF). Fluorescent in situ hybridization (FISH) is a powerful molecular method for the specific and rapid diagnosis of bacteria, including the detection of P. aeruginosa in sputum samples from CF patients. High background fluorescence of viscous sputum samples obtained from CF patients may impede detection of microorganisms by FISH. The aim of this study was to test the application of biotin during FISH technique to reduce unspecific background fluorescence in sputum samples to facilitate and improve detection of P. aeruginosa. Sixty-three sputum samples from CF patients were tested by FISH to detect P. aeruginosa. All the 63 samples were also examined by a modified FISH procedure including biotin treatment. The FISH results were compared with those of conventional culture method. The specificity of FISH was 100%. The sensitivity of FISH for detection of P. aeruginosa from samples without biotin treatment was 83.3%, whereas in biotin-treated samples was 88.1%. Biotin reduced background fluorescence of 12 sputum samples of CF patients and it did not show any adverse effect on FISH results of the remaining sputum samples. Therefore, using of biotin in FISH procedure seems to facilitate and improve the detection of respiratory tract infections by P. aeruginosa in this population

Antibacterial activity of indium curcumin and indium diacetylcurcumin

Studies on curcumin, the principal element of turmeric powder, have demonstrated several biological actions such as antibacterial activity. Evaluation of new analogs or new compounds of curcumin for their antibacterial effect is interesting for researchers. In this in vitro study, we attempted to test the antibacterial activity of indium curcumin (In(CUR)3), indium diacetylcurcumin (In(DAC)3), and diacetylcurcumin (DAC) in comparison with curcumin. The action of these agents were examined on Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (ATCC 14990), Pseudomonas aeruginosa (ATCC 27853), and Escerichia coli (ATCC 25922). Curcumin was effective against S. aureus and S. epidermidis, whereas In(DAC)3 showed activity against S. epidermidis and P. aeruginosa. The effect of In(DAC)3 on P. aeruginosa is an advantage. Strikingly, In(CUR)3 exhibited antibacterial activity on all the four mentioned strains. DAC did not show antibacterial effect on any of the four test bacteria. The minimum inhibitory concentration (MIC) of curcumin was 187.5 μg/ml for S. aureus, and 46.9 μg/ml for S. epidermidis. However, the MIC of In(CUR)3 was lower for the same bacterial strains (93.8 μg/ml for S.aureus and 23.4 μg/ml for S. epidermidis). Therefore, In(CUR)3 was found to have more antibacterial effect than curcumin itself and could be a suitable candidate for further in vivo investigations

Invitro Antibacterial Activity of the Prosopis Juliflora Seed Pods on Some Common Pathogens

Introduction: Prosopis juliflora is probably the most widespread species of genus Prosopis and it is a good source of compounds that have been shown to be pharmacologically active. This plant has been used as a traditional treatment for several diseases. Aim: To investigate the in-vitro antibacterial activity of the P. juliflora seed pods from Bushehr, South West of Iran. Materials and Methods: In the present study, the antibacterial activity of P. juliflora seed pods extract was tested against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Pseudomonas aeruginosa. The minimum inhibitory concentration (MIC) of the extract was determined for each test microorganism. Results: P. juliflora seed pods extract exhibited antibacterial activity against all four test organisms. The MIC of the extract was 0.312 mg/ml and 0.078 mg/ml for S. aureus and S. epidermidis, respectively and 1.25 mg/ml for both E.coli and P.aeruginosa. Conclusion: P. juliflora seed pods from Bushehr, South West of Iran could be an appropriate source of antibacterial compounds that makes it a promising candidate for further studies

Evaluation of fluorescent in situ hybridization for rapid diagnosis of enterococcal wound infection

Enterococci are among prominent causes of nosocomial wound infections. Since the rapid detection of causative agents could make earlier administration of choice antibiotics and quick recovery of patients, so the application of rapid diagnostic methods is important. Therefore, this study was designed to evaluate fluorescent in situ hybridization (FISH) for the detection of Enterococcus in wound swab samples. The time needed for FISH procedure is about 3 h. Specimens taken from 33 hospitalized patients were examined by both FISH and culturing procedures. By using conventional culture, 10 of 33 wound samples were culture-positive. Out of these 10 specimens, eight were FISH-positive, but two specimens were FISH-negative for Enterococcus. The remaining 23 wound specimens were Enterococcus negative according to the both methods. Therefore, the specificity of FISH was 100%; however, this method showed 80% sensitivity. Because of high specificity of FISH, the combined application of FISH and cultivation methods would be suggested for detection of enterococci from wound specimens in situations in which rapid diagnosis has an advantage in the treatment of patients

In vitro antitumor activity of Gracilaria corticata (a red alga) against Jurkat and molt-4 human cancer cell lines

Gracilaria corticata is a red alga which can be collected from many sea coasts around the world such as China, India, Persian Gulf, etc. The Persian Gulf is a unique marine habitat infested with diverse seaweeds. The aim of the present study is to explore anticancer potential of the crude extracts from G. corticata which was collected from the Bushehr coast (South west of Iran). Here, different concentration of the aqueous extract from G. corticata was tested for probable antitumoral activity on Jurkat and molt- 4 human lymphoblastic leukemic cell lines. The cells were treated by different concentration of algal extract and the number of viable cells was determined by trypan blue. Also, cytotoxicity of the extract was evaluated by methyl thiazolyl tetrazolium (MTT) assay. The results showed that 9.336 and 9.726 μg/μl of algal extract were the most effective concentrations against Jurkat and molt-4 cells, respectively. The water crude extract of red alga G. corticata had significant anticancer activity and it might be a good candidate for further investigations in order to develop a natural compound as an anticancer agent which can be used for the production of potential anticancer drug and novel pharmaceutical leads

USE OF A MODIFIED FLUORESCENT IN SITU HYBRIDIZATION PROCEDURE TO IMPROVE THE IDENTIFICATION OF STREPTOCOCCUS PNEUMONIAE IN BLOOD CULTURES

Streptococcus pneumoniae is an important causative agent for bacteremia. Fluorescent in situ hybridization (FISH) is a helpful molecular technique for the rapid identification of S. pneumoniae in positive blood cultures. There are many reports concerning the application of an enzymatic treatment with lysozyme in the FISH procedure for partial cell wall digestion of S. pneumoniae. However, this study was aimed to test the FISH procedure without enzymatic treatment for the identification of S. pneumoniae in blood culture specimens. Seventy-seven positive blood culture specimens containing Gram-positive cocci were examined by both the conventional laboratory methods and FISH. Detection of S. pneumoniae was performed by two FISH procedures: one procedure was performed with an enzymatic treatment step and the other one was done without enzymatic treatment. In addition, the specimens were tested by the FISH procedure with enzymatic treatment to detect Streptococcus pyogenes and Enterococcus. The specificity of FISH in comparison with conventional culture methods was 100%. The sensitivity of the FISH procedure with enzymatic treatment for the detection of S. pneumoniae was 90%, whereas, the sensitivity of the FISH procedure without enzymatic treatment was 100%. In fact, by omission of enzymatic treatment, detection of S. pneumoniae was improved in 6 specimens. The results of the FISH and culture methods for the detection of S. pyogenes and Enterococcus were compatible. Altogether, FISH procedure without enzymatic treatment step seems to improve the detection of S. pneumoniae in some case

Seroepidemiological study of toxoplasmosis in childbearing age women in Bushehr City, south west of Iran in 2009

Toxoplasmosis is a parasitic disease caused by an intracellular protozoan, Toxoplasma gondii and congenital form is one of the most important clinical aspects of this disease. Seroepidemiological studies among women of childbearing age could provide appropriate approaches to design prevention measures to prevent congenital Toxoplasmosis. In this study, 303 blood samples were collected from women of childbearing age who referred to the health center clinics for premarital examinations in Bushehr city. Sera were tested for anti-Toxoplasma IgG and IgM antibodies by using ELISA method. 71 cases (23.4%) were seropositive and 232 (76.6%) serum samples were seronegative for anti-Toxoplasma IgG antibody, 10 samples (3.3%) were seropositive and 293 samples (96.7%) were seronegative for antiToxoplasma IgM antibody. According to the results, there was no significant association between seropositivity and age groups, area of residency, history of contact with cat, educational levels and nutritional behaviors. Most women of childbearing age in Bushehr City do not have immunity against Toxoplasmosis. These women are at risk of acquired Toxoplasmosis in their pregnancy period. It is recommended to health managers to design the educational measures to prevent congenital Toxoplasmosis

Seroepidemiological study of toxoplasmosis in childbearing age women in Bushehr City, south west of Iran in 2009

Toxoplasmosis is a parasitic disease caused by an intracellular protozoan,Toxoplasma gondii and congenital form is one of the most important clinical aspects of this disease. Seroepidemiological studies among women of childbearing age could provide appropriate approaches to design prevention measures to prevent congenital Toxoplasmosis. In this study, 303 blood samples were collected from women of childbearing age who referred to the health center clinics for premarital examinations in Bushehr city. Sera were tested for anti-Toxoplasma IgG and IgM antibodies by using ELISA method. 71 cases (23.4%) were seropositive and 232 (76.6%) serum samples were seronegative for anti-Toxoplasma IgG antibody, 10 samples (3.3%) were seropositive and 293 samples (96.7%) were seronegative for anti-Toxoplasma IgM antibody. According to the results, there was no significant association between seropositivity and age groups, area of residency, history of contact with cat, educational levels and nutritional behaviors. Most women of childbearing age in Bushehr City do not have immunity against Toxoplasmosis. These women are at risk of acquired Toxoplasmosis in their pregnancy period. It is recommended to health managers to design the educational measures to prevent congenital Toxoplasmosis.Keywords: Toxoplasma gondii, premarital women, seroepidemiolog, ELISAAfrican Journal of Biotechnology Vol. 9(36), pp. 5809-5812, 6 September, 201