Characterization of the biosurfactant production and enzymatic potential of bacteria isolated from an oil-contaminated saline soil / Caracterización de la producción de biosurfactantes y potencial enzimático de bacterias aisladas de un suelo salino contaminado con aceite

Biosurfactants are amphiphilic compounds with extensive applications in oily contaminated environments to remove hydrocarbons. Moreover, enzymes such as laccase and manganese peroxidase are responsible for the oxidation of a variety of phenolic compounds and aromatic amines. Therefore, in the present study, bacteria with the potential to produce biosurfactants and enzymes (namely, laccase, manganese peroxidase, and endoglucanase carboxymethyl cellulose (CMCase)) were isolated from petroleum oil-contaminated soil. From 15 isolated bacteria, three isolates were selected as the best producers of biosurfactants according to the related tests, such as tests for surface tension reduction. These three bacteria indicated tolerance to a salinity test and were classified as resistant and very resistant. The isolates 3, 12, 13, and 14 showed positive results for the degradation of guaiacol, phenol red, and carboxymethylcellulose, as well as the decoloration of methylene blue by the creation of a clear halo around the bacterial colony. Upon the quantitation of the laccase and manganese peroxidase activities, 22.58 U/L and 21.81 U/L, respectively, were measured by isolate 13. Furthermore, CMCase activity was recorded with 0.057436 U/ml belonging to isolate 14. Bacterial strains with appreciable laccase, peroxidase, CMCase activity, and biosurfactant production potentials were identified through 16S rDNA sequence analysis as Bacillus sp. (isolate 3), Bacillus toyonensis (isolate 12), Bacillus cereus (isolate 13), and Bacillus tropicus (isolate 14), and their nucleotide sequences were deposited in the GenBank. The potentials for the industrial applicability of the biosurfactants and enzymes abound, and production needs to be optimized by the selected bacterial strains.(AU)

Characterization of the biosurfactant production and enzymatic potential of bacteria isolated from an oil-contaminated saline soil.

Biosurfactants are amphiphilic compounds with extensive applications in oily contaminated environments to remove hydrocarbons. Moreover, enzymes such as laccase and manganese peroxidase are responsible for the oxidation of a variety of phenolic compounds and aromatic amines. Therefore, in the present study, bacteria with the potential to produce biosurfactants and enzymes (namely, laccase, manganese peroxidase, and endoglucanase carboxymethyl cellulose (CMCase)) were isolated from petroleum oil-contaminated soil. From 15 isolated bacteria, three isolates were selected as the best producers of biosurfactants according to the related tests, such as tests for surface tension reduction. These three bacteria indicated tolerance to a salinity test and were classified as resistant and very resistant. The isolates 3, 12, 13, and 14 showed positive results for the degradation of guaiacol, phenol red, and carboxymethylcellulose, as well as the decoloration of methylene blue by the creation of a clear halo around the bacterial colony. Upon the quantitation of the laccase and manganese peroxidase activities, 22.58 U/L and 21.81 U/L, respectively, were measured by isolate 13. Furthermore, CMCase activity was recorded with 0.057436 U/ml belonging to isolate 14. Bacterial strains with appreciable laccase, peroxidase, CMCase activity, and biosurfactant production potentials were identified through 16S rDNA sequence analysis as Bacillus sp. (isolate 3), Bacillus toyonensis (isolate 12), Bacillus cereus (isolate 13), and Bacillus tropicus (isolate 14), and their nucleotide sequences were deposited in the GenBank. The potentials for the industrial applicability of the biosurfactants and enzymes abound, and production needs to be optimized by the selected bacterial strains.

Simple synthesis of novel magnetic silver polymer nanocomposites with a good separation capacity and intrinsic antibacterial activities with high performance.

Two new coordination polymers namely, [(AgCN)4LS]n (1) and [(AgCN)3LN]n (2), were successfully synthesized by the reaction of AgNO3 and cyanide as a co-anion with LS[1,1'-(hexane-1,4-diyl)bis(3-methylimidazoline-2-thione] and LN[1,1,3,3-tetrakis(3,5-dimethyl-1-pyrazole)propane] ligands in order to use them for the preparation of magnetic nanocomposites with MnFe2O4 nanoparticles by an efficient and facile method. They were then well characterized via numerous techniques, including elemental analysis, FT-IR spectroscopy, TGA, PXRD, SEM, TEM, EDX, VSM, BET, ICP, and single-crystal X-ray diffraction. The considered polymers and their magnetic nanocomposites with nearly the same antibacterial activity demonstrated a highly inhibitive effect on the growth of Gram-negative (Escherichia coli, Pseudomonas aeruginosa) and Gram-positive (Staphylococcus aureus, Bacillus subtilis) bacteria. By considering the simple separation and recyclable characters of the magnetic nanocomposites, these materials are suitable to be used in biological applications.

Highly sensitive determination of methionine by solvent-based de-emulsification dispersive liquid-liquid microextraction using bio-stabilized silver nanoparticles.

In this study, a solvent-based de-emulsification dispersive liquid-liquid microextraction method coupled with surface plasmon resonance of silver nanoparticles was developed for determination of trace levels of methionine. The stable and dispersed silver nanoparticles were synthesized by applying ascorbic acid as reducer and Stenotrophomonas sp. bacterial suspension as bio-stabilizer and then preconcentrated in organic phase according to a facile dispersive liquid-liquid microextraction procedure based on 1-octanol as extraction solvent, methyltrioctylammonium chloride (aliquat 336) as disperser and acetone as de-emulsifier. The presence of methionine influenced the intensity of plasmon resonance absorbance of silver nanoparticles, which was employed as a colorimetric probe for the determination of this amino acid. Under the optimal conditions, the linear analytical range of 5.6 to 234.5 nmol/L and a detection limit of 3.4 nmol/L were achieved for methionine. The relative standard deviation for seven replicate measurements of 33.5 and 107.2 nmol/L of methionine was 4.3 and 2.1%, respectively. The suggested method was successfully applied for the determination of methionine in biological samples.

Environmentally friendly acetic acid/steam explosion/supercritical carbon dioxide system for the pre-treatment of wheat straw.

It is well established that pretreatment of lignocellulosic biomass is required to achieve an effective enzymatic saccharification process. At the present time, most of the touted pre-treatment technologies would cause environmental pollution and unsustainable water use for the pretreated material prior to enzymatic saccharification. To address these shortcomings, the pretreatment technology which combines the supercritical CO2, SC-CO2 (a green solvent), acetic acid, and steam explosion was used to assess the pretreatment of wheat straw for enzymatic saccharification. The effects of solvent concentration, impregnation temperature and time, pre-treatment time, and temperature, as well as SC-CO2 pressure, contact time, and temperature, were evaluated. The results identified that at the optimum SC-CO2 pressure of 18 MPa, the highest amount of reducing sugars (RS) was produced from the cellulosic pulp using Acetic acid/Steam/SC-CO2 at 200 °C for 30 min, a value 20% more than the pulp produced with the Water/Steam/SC-CO2. The effectiveness of the pretreatment process was attributed not only to delignification and defibrillation but also to the exposure of the cellulose structure evidenced from the proportion of the ß-glycosidic linkages as shown by FTIR. Passing SC-CO2 after the pretreatment reduces the amounts of fermentation inhibitors and eliminates the use of wash water.

Screening Heterotrophic Ammonia Removal and Aerobic Denitrifying Bacteria from Wastewater of Ammonia Production Units of a Petrochemical Industry.

Petrochemical wastewaters have high ammonium contents that can cause serious environmental pollutions. Among different treatment methods, biological ammonia removal using bacteria has been attracted more attention due to their environmental friendly and high efficiency. The aim of the present study was to isolate heterotrophic nitrogen removal and aerobic denitrifying bacteria from petrochemical wastewater to be used for bioremediation. Wastewater and sludge samples were prepared and their chemical parameters were analyzed. Bacterial isolation was done through enrichment in mineral salt medium and basal salt medium and identified through 16S rRNA gene sequencing. The ability of isolates was evaluated at different concentrations of ammonia, nitrate, and carbon sources and at different time intervals. As a result, ten bacterial isolates were obtained that 5 strains were identified as heterotrophic nitrogen removal bacteria and 5 strains were able to do aerobic denitrification. From these, Pseudomonas guguanensis strain 4-n-1 with 93.2% ammonia removal during 24 h at 40 mg/L ammonium sulfate concentration and Pseudomonas guariconensis strain 5-d-1 with 89.2% denitrification during 24 h at 100 mg/L nitrate concentration were the best isolates. Finally, based on the efficiency of these isolates it can be concluded that effective ammonia removal can be achieved using these species as consortium and simultaneous heterotrophic nitrogen removal with aerobic denitrification is suitable method for efficient ammonium removal from petrochemical wastewater and reduction of ammonia concentration.

Effect of Various Isolated Microbial Consortiums on the Biodegradation Process of Precipitated Asphaltenes from Crude Oil.

One of the serious problems in the oil industry is precipitation and deposition of asphaltenes in the different oil production stages including formation, wellbore, production tubing, flow lines, and separation units. This phenomenon causes a dramatic increase in the cost of oil production, processing, and transferring. Thus, it seems to be very necessary to use the removing methods for precipitated asphaltenes in different crude oil production and transferring stages. In this study, the ability of microorganisms for biodegradation of precipitated asphaltenes was investigated. For this purpose, four bacterial consortiums were isolated from oil-contaminated soil, crude oil, reservoir water, and oil sludge samples of an oil field located in the southwest of Iran. Based on the results of the designed experiments, by using response surface methodology (RSM) and central composite design, the bacterial consortiums were cultured in the flasks. Three levels of temperatures, salinity, pH, and initial asphaltene concentration as the substrate were considered as the parameters of culture medium and incubated growth mediums for 60 days. The maximum asphaltene biodegradation was 46.41% caused by the crude oil consortium including Staphylococcus saprophyticus sp. and Bacillus cereus sp. at 45 °C, salinity 160 g·L-1, pH 6.5, and 25 g·L-1 initial asphaltene concentration. Also, it was observed that the negative or positive impacts of culture media conditions such as temperature and salinity on asphaltene degradation depended on the type of the available bacterial consortium. The carbon-hydrogen-nitrogen-sulfur analysis showed that carbon, hydrogen, nitrogen, and in some cases, the sulfur in biodegraded samples are less than in control samples. Moreover, Fourier transform infrared analysis indicated that the alkyne groups were less resistant to biodegradation and were eliminated thoroughly after 2 months of incubation. In addition, alkane components were partially removed in treated asphaltene fraction. The parameters of culture medium were optimized by RSM, and besides, their effects on the performance of bacteria in the asphaltene biodegradation process were discussed. The validity of some available kinetic models to describe the behavior of the studied bacteria consortium was investigated, and it was observed that Tessier, Moser, and Contois models accurately predict the values of asphaltenes and biomass concentration at 30, 45, and 60 °C, respectively.

Co-composting of oil-based drilling cuttings by bagasse.

The feasibility of using a locally abundant bulking material (sugarcane bagasse) in Khuzestan province, Iran, to remove petroleum hydrocarbons from oil-based drill cuttings (OBDCs), using composting process, was investigated. OBDCs were collected from the discharge point of a drilling rig and bagasse was collected from a sugarcane agro-industry near Ahwaz. Experiments were performed in the dark and at room temperature, using different bagasse to OBDCs ratios. Degradation extent and kinetics of total petroleum hydrocarbons (TPHs) and poly aromatic hydrocarbons (PAHs), as well as dehydrogenase and urease enzymes activities and number of heterotroph bacteria during the co-composting of OBDCs and bagasse were studied and measured. Highest PAHs and TPH removals were observed when OBDCs were composted with 15% bagasse. After 70 days of incubation, PAHs and TPH were removed up to 24.8% and 67.5%, respectively. Studying the enzymes activities and number of heterotrophs with TPH and PAHs concentrations over time suggests that biodegradation is the main mechanism in the degradation process. The first-order kinetic model was fitted to the TPH and PAHs degradation data and contaminants half-lives were estimated to be in the range of 40-80 and 170-240 days respectively. DT90 values for TPH and PAHs were in the range of 120-260 and 560-1260 days, depending on the bagasse content.

Fibrinolysin production by Alcaligenes faecalis strain 26 isolated from environment.

BACKGROUND AND OBJECTIVES: Fibrinolytic drugs are commonly used for fibrin clot lysis but due to their inappropriate side effects, as well as their high costs, using fibrinolytic enzymes has been paid attention. Bacterial sources of this enzyme are a good alternative for this purpose. The aim was fibrinolysin production through screening of fibrinolysin producing bacteria from environmental samples. MATERIALS AND METHODS: Bacterial isolation was performed from different environmental samples and was screened based on sheep blood clot digestion and culture on plasma plate. The most potent isolate was optimized for different growth parameters including temperature, pH and fibrinolysin production at optimum growth conditions. The stability of produced enzyme at various temperatures and pH and treatment with MgSO4, NiSO4, SDS and EDTA was then investigated. Finally this isolate was identified based on the 16S rRNA sequencing. RESULTS: As a result, from 79 different isolates, the most potent fibrinolysin producer was identified as Alcaligenes faecalis strain 26. This isolate produced 12 mm halo zone on plasma plate. Its optimum growth temperature and pH was 43°C and 7, respectively. The produced enzyme had the best stability at pH 7 and was also active up to 60°C. The fibrinolytic activity of this isolate was reduced following treatment with MgSO4, NiSO4 and also with protease inhibitors, such as SDS and EDTA. CONCLUSION: Based on the obtained results it can be suggested that Alcaligenes faecalis strain 26 has appropriate efficiency for fibrinolysin production that can be used in food industry and medicine.

Diversity and metallo-ß-lactamase-producing genes in Pseudomonas aeruginosa strains isolated from filters of household water treatment systems.

The microbiological quality of drinking water has long been a critical element in public health. Considering the high clinical relevance of Pseudomonas aeruginosa, we examined the filters of household water treatment systems for its presence and characteristics to determine the systems' efficiency in eliminating the bacteria. In total, filters of 50 household water treatment systems were examined. Microbiological and molecular methods were used for the detection and confirmation of P. aeruginosa isolates. Random Amplification of Polymorphic DNA-polymerase chain reaction (RAPD-PCR) was performed to detect similarities and differences among P. aeruginosa isolates. Combined disk (CD) method and double disk synergy test (DDST) were performed to detect metallo-beta-lactamase (MBL)-producing P. aeruginosa isolates. Finally, PCR was performed to detect MBL genes in MBL-producing strains. From the 50 analyzed systems, 76 colonies of P. aeruginosa were identified. In some systems, isolated bacteria from different filters harbored similar genetic profiles, indicating that these isolates may be able to pass through the filter and reach higher filters of the system. Phenotypic tests revealed 7 (9.2%) MBL-producing strains. Two isolates were positive for blaVIM-1, whereas one isolate was positive for blaNDM and blaIMP-1. The wide distribution of resistant phenotypes and genetic plasticity of these bacteria in household water treatment systems indicate that resistance mechanisms circulate among P. aeruginosa isolates in the environment of the filtration systems. The presence of MBL-producing genes in these systems and P. aeruginosa as a potential reservoir of these resistance genes can be a major concern for public health.

Ecofriendly demulsification of water in oil emulsions by an efficient biodemulsifier producing bacterium isolated from oil contaminated environment.

OBJECTIVE: Water in oil emulsions increase oil processing costs and cause damage to refinery equipment which necessitates demulsification. Since chemical demulsifiers cause environmental pollution, biodemulsifiers have been paid more attention. This study aims to identify biodemulsifier-producing bacteria from petroleum contaminated environments. RESULTS: As a result, several biodemulsifier producing strains were found that Stenotrophomonas sp. strain HS7 (accession number: MF445088) which produced a cell associated biodemulsifier showed the highest demulsifying ratio, 98.57% for water in kerosene and 66.28% for water in crude oil emulsion after 48 h. 35 °C, pH 7, 48 h incubation and ammonium nitrate as nitrogen source were optimum conditions for biodemulsifier production. Furthermore, it was found that hydrophobic carbon sources like as liquid paraffin is not preferred as the sole carbon source while a combination of various carbon sources including liquid paraffin will increase demulsification efficiency of the biodemulsifier. CONCLUSIONS: The appropriate potential of this biodemulsifier strengthens the possibility of its application in industries especially petroleum industry.

Magnetic graphene oxide inlaid with silver nanoparticles as antibacterial and drug delivery composite.

A three-component composite consisted of graphene oxide, cobalt ferrite, and silver nanoparticles has been prepared by a facile method and fully characterized. The antibacterial activity of this composite has been greatly enhanced after being combined with ciprofloxacin drug. This clearly showed the occurrence of a strong synergistic effect between ciprofloxacin and the Ag NPs in the composite. The ciprofloxacin-conjugated composite was found to be a potent antimicrobial agent while having rather low cytotoxicity and high stability. The studies based on field emission scanning electron microscopy (FESEM) analysis and zeta potential measurement have revealed that the composite sticks to the bacterial cell wall causing irreversible cell damage. This multifunctional magnetic nanocomposite was also examined as drug delivery system for ciprofloxacin in solutions with different pH. It was observed that the release of ciprofloxacin in this system is pH-sensitive with gradual and controlled manner. Mechanisms for the synergistic effect and drug release behavior, as well as explanation for the antibacterial action, of the nanocomposite were also demonstrated.

Synthesis, structural characterization, QSAR and docking studies of a new binuclear nickel (II) complex based on the flexible tetradentate N-donor ligand as a potent antibacterial and anticancer agent.

A new nickel (II)complex namely [Ni2(Lt)Cl4] derived from the NiCl2.6H2O and 1,1,3,3-tetrakis(3,5-dimethyl-1-pyrazolyl)propane (Lt) has been synthesized and fully characterized by the single crystal X-ray diffraction, elemental analysis, FT-IR, UV-vis, density functional theory (DFT) calculations, antibacterial and anticancer activities. In the title complex, each of the Ni(II) atoms is tetrahedrally coordinated by two N atoms from one of the chelating bidentate bis(3,5-dimethylpyrazolyl)methane units of the Lt ligand and two Cl as terminal ligands. The neighboring [Ni2(Lt)Cl4] molecules are linked together by the intermolecular CH⋯Cl hydrogen bonds to generate a 1D chain structure. The chains are further stabilized by the intermolecular CH⋯π interactions to form a two-dimensional non-covalent bonded structure. The antibacterial activity of the free Lt ligand and its Ni (II) complex shows that the ability of these compounds to inhibit growth of the tested bacteria increase from the Lt to binuclear nickel (II) complex. Scanning probe microscopy (SPM) study of the treated B. subtilis and E. coli bacteria was implemented to understand the structural changes caused by the interactions between the nickel (II) complex and the target bacteria. The cytotoxicity test of the Lt ligand and its complex was evaluated against the human carcinoma cell line (Caco-2) using the MTT assay. The results indicate that the Lt ligand and its complex display cytotoxicity against Caco-2 with the IC50 values of 36.29µM and 12.97µM, respectively. Therefore, the complex can be nominated as a potential anticancer agent. Molecular docking investigations on the five standard antibiotic, five standard anticancer drugs, free Lt ligand, title complex and twelve receptors were performed by Autodock vina function. The results of docking and DFT calculations are in line with the in vitro data obtained via the antibacterial and anticancer activity of Lt ligand and its made-complex.

Biological Synthesis of Selenium Nanoparticles and Evaluation of their Bioavailability

ABSTRACT Nanoparticles due to their unique properties have attracted more attention and their bacterial biosynthesis is more favorable because is environmental friendly and the size and yield of nanoparticles can be optimized. The aim of the present study was biosynthesis of Selenium nanoparticles using Bacillus cereus. For this purpose, bacterial culture was prepared in the presence of sodium selenate solution and incubated (30°C, 24 h). The produced nanoparticles were purified through consequent centrifugation, washing with 0.9% NaCl, sonication, washing with Tris- HCl containing Sodium dodecyl sulfate (SDS) and finally isolation with water- octanol two phase systems. Then using Ultraviolet-Visible spectroscopy, dynamic light scattering (DLS), scaning electron microscopy (SEM) and X-ray diffraction (XRD) analysis, nanoparticle production was confirmed. The bioavailability of nanoparticles was also investigated in rat. As a result of this study spherical selenium nanoparticles with a mean diameter of 170 nm were biosynthesized. MIC (minimum inhibitory concentration) and MBC (minimum bactericidal concentration) of selenium for Bacillus cereus were same and equal to 75mM. Absorption and secretion of nanoselenium was significantly higher than bulk Selenium (P<0.05). In conclusion in the present study without any chemical substance, spherical Selenium nanoparticles were produced that do not have any environmental contamination. Furthermore, the metabolism of these particles suggests higher absorption rate of them that facilitates its application in medicine and also veterinary medicine.

Phylogenetic typing and detection of extended-spectrum ß-lactamases in Escherichia coli isolates from broiler chickens in Ahvaz, Iran.

This study was conducted to reveal the phylogenetic background, to detect the genes encoding TEM, SHV and CTX-M-15 extended-spectrum ß-lactamases (ESBL), and to analyze their distribution in phylo-groups of 150 Escherichia coli isolates from broiler chickens in Ahvaz (Southwest of Iran). Seventy- five cloacal swabs from healthy birds (fecal isolates), and 75 heart blood samples from birds with colibacillosis (septicemic isolates) were obtained. All isolates were phylotyped and screened for ESBL genes by polymerase chain reaction (PCR). The fecal isolates belonged to four main phylo-groups, including 41 isolates (54.67%) to A, 9 (12.00%) to B1, 5 (6.67%) to B2, and 20 (26.67%) to D. Of septicemic isolates, 37 isolates (49.33%) were classified as phylotype A, 5 (6.67%) as B1, 10 (13.33%) as B2, and 23 (30.67%) as D. In molecular analysis, a total of 72 isolates (35 fecal and 37 septicemic) were identified to harbor ESBL genes, which were distributed in phylo-groups A, B1, B2, and D. Regardless of the type of isolate, blaCTX-M-15 gene was the most common genotype, followed by blaTEM and blaSHV genes. This study suggests that broiler chickens in Iran are infected to ESBL genes- harboring Escherichia coli strains which may be spread to the food chain through fecal contamination of carcasses during slaughtering.

Molecular Detection and Typing of Human Papillomaviruses in Paraffin-Embedded Cervical Cancer and Pre-Cancer Tissue Specimens.

BACKGROUND: Cervical cancer is one of the important reasons of mortality among females. Prevention, early diagnosis and immediate treatment can affect the rate of mortality in this cancer and several epidemiological studies have shown a strong relationship between human papilloma viruses (HPVs) and cervical cancer. OBJECTIVES: The present study was conducted to survey HPV infections in a women population with cervical cancer and cervical dysplasia/metaplasia in southwest of Iran. MATERIALS AND METHODS: 72 paraffin-embedded cervical biopsies which had been previously archived from women with cervical cancer and cervical dysplasia were examined by polymerase chain reaction (PCR). Afterward, the detected HPV strains were typed by restriction fragment length polymorphism (RFLP) analysis of PCR amplicons. RESULTS: 60 out of 72 samples had necessary requirements and HPV DNA was detected in 43.3% of these samples. Most HPV positive samples belonged to women aged from 48 to 63 years. On the other hand, HPV infection among patients with squamous cell carcinoma (SCC) was 48.78% and in women with dysplasia/metaplasia was 26.66%. The most prevalent type of the human papilloma virus was HPV16 (100%). CONCLUSIONS: Knowing the most prevalent type of the human papilloma viruses circulating in the population (HPV16) can be applied in the future screening and managing programs of this major disease and also in vaccination against the prevalent types of the virus. Meanwhile, it seems that more studies should be performed to determine the role of different risk factors involved in development of the disease, especially those related with social behaviors and traditions with respect to different areas.

The Association of Panton-Valentine leukocidin and mecA Genes in Methicillin-Resistant Staphylococcus aureus Isolates From Patients Referred to Educational Hospitals in Ahvaz, Iran.

BACKGROUND: Staphylococcus aureus, an important human pathogen is one of the main causative agents of nosocomial infection. Virulence genes play a major role in the pathogenicity of this agent and its infections. Methicillin-Resistant Staphylococcus aureus (MRSA) isolates are major challenge among infectious agents that can cause severe infections and mortality. Methicillin-resistant S. aureus produces a unique type of Penicillin Binding Protein 2a (PBP2a) that has low affinity for ß-lactam antibiotics. Most of the MRSA bacterial strains can also produce a leukotoxin as Panton-Valentine Leukocidin (PVL) that increases the virulence of MRSA strains and can cause severe necrotic pneumonia. The presence of pvl gene is a genetic marker for the MRSA populations. OBJECTIVES: The aim of this study was to explore the association of pvl and mecA genes in clinical isolates of MRSA. MATERIALS AND METHODS: Fifty MRSA isolates were collected from 200 clinical samples from three different educational hospitals in Ahvaz, Iran, and identified by biochemical tests including catalase, oxidase, tube coagulase, mannitol fermentation, and sensitivity to furazolidone, resistance to bacitracin, PYR test and Voges-Proskauer test. Their resistance to methicillin was evaluated using the disc diffusion method. DNA was extracted by boiling and then the presence of pvl and mecA genes was investigated by the polymerase chain reaction method using specific primers. RESULTS: The results revealed that mecA and pvl genes were positive for 15 (30%) and 3 (6%) of the isolates, respectively. None of mecA positive isolates was positive for pvl gene. CONCLUSIONS: It can be concluded from these results that fortunately the prevalence of pvl gene is low in MRSA isolates in this region and there is no association between the presence of pvl and mecA genes in these isolates.

Production of Polyhydroxybutyrate by Bacillus axaraqunsis BIPC01 using Petrochemical Wastewater as Carbon Source

The aim of this study was to use petrochemical wastewater as the source of carbon for the production of polyhydroxyalkanoates (PHA) in an effort to decrease its cost of production. For this purpose, PHA producing bacteria were isolated from the petrochemical wastewater of Bandar Imam, Iran. The purified colonies were screened for PHA by Sudan Black B and Nile Blue A staining. Among positively stained bacteria, the best PHA producer was selected on the basis of cell growth, PHA content and the monomer composition of PHA. The phenotypic and genotypic identification this isolate showed it to be Bacillus axaraqunsis. The PHA was produced at a cell density of about 9.46 g/l of maximum concentration of 6.33g/l l, corresponding to 66% of cell dry weight. These results showed that B. axaraqunsis BIPC01 could be a potent PHA producer using wastewater for industrial purpose and simultaneously reducing the environmental pollution.

The Antibacterial Activity of Cassia fistula Organic Extracts.

BACKGROUND: Cassia fistula, is a flowering plant and a member of Fabaceae family. Its leaves are compound of 4 - 8 pairs of opposite leaflets. There are many Cassia species around the world which are used in herbal medicine. OBJECTIVES: This study was designed to examine in vitro anti-bacterial activity of methanolic and ethanolic extracts of C. fistula native to Khuzestan, Iran. MATERIALS AND METHODS: The microbial inhibitory effect of methanolic and ethanolic extracts of C. fistula was tested on 3 Gram positive: Bacillus cereus, Staphylococcus aureus and S. epidermidis and 5 Gram negative: Salmonella Typhi, Kelebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, and Proteus mirabilis bacterial species using disc diffusion method at various concentrations. The minimum inhibitory and bactericidal concentrations (MIC and MBC) were measured by the tube dilution assay. RESULTS: The extract of C. fistula was effective against B. cereus, S. aureus, S. epidermidis, E. coli and K. pneumoniae. The most susceptible microorganisms to ethanolic and methanolic extracts were E. coli and K. pneumoniae, respectively. Also B. cereus and S. aureus showed the least sensitivity to ethanolic and methanolic extracts, respectively. The MIC (minimum inhibitory concentration) and MBC (minimum bactericidal concentration) of ethanolic extracts against S. aureus, E. coli, S. epidermidis and K. pneumoniae were also determined. CONCLUSIONS: With respect to the obtained results and regarding to the daily increase of the resistant microbial strains to the commercial antibiotics, it can be concluded that these extracts can be proper candidates of antibacterial substance against pathogenic bacterial species especially S. aureus, E. coli, K. pneumoniae and S. epidermidis.

Introducing Urtica dioica, A Native Plant of Khuzestan, As an Antibacterial Medicinal Plant.

BACKGROUND: Urtica dioica is a flowering plant with long history of use in folk medicine and as a food source. OBJECTIVES: This study examined in vitro antibacterial potential of alcoholic extracts of U. dioica. MATERIALS AND METHODS: Hydroalcoholic extracts from aerial parts were prepared using aqueous solution of ethanol and methanol and their inhibitory effects against clinical isolates was examined by disc diffusion method at different doses. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentration (MBC) indexes were also investigated. The scanning electron microscopy (SEM) analysis was also performed to find structural changes of affected bacteria consequent to exposing with extracts. RESULTS: Both extracts were active against Bacillus cereus, Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli with respectively 16, 10, 18, and 14 mm (methanolic) and 11, 9, 17, and 16 mm (ethanolic) inhibition zone. The MIC of ethanolic extract against S. epidermidis and E. coli was respectively 10 and 40 mg/mL. The MIC of methanolic extract against S. aureus and S. epidermidis was 40 and 10 mg/mL, respectively. The MBC was found only for S. epidermidis (20 mg/mL). In SEM analysis the round shape of S. epidermidis was changed and irregular shapes were appeared, which suggest that the main target of these extracts was cell wall. CONCLUSIONS: Extracts of U. dioica showed significant antibacterial effect against some clinically important pathogenic bacteria. Based on the obtained results it can be concluded that U. dioica is useful as antibacterial and bactericidal agent in treating infectious diseases.