The Effect of Lime Stabilization on E. coli Destruction and Heavy Metal Bioavailability in Sewage Sludge for Agricultural Utilization

The addition of lime as Ca(OH)2 to sewage sludge to destroy pathogens (Escherichia coli), was evaluated also in relation to heavy metal bioavailability. The obtained results show that the use of calcium hydroxide at the dose of 3% effectively destroyed pathogens ensuring the stability at high pH values over long period and the duration of the sewage sludge stabilization. In general, lime addition decreased the total extractability of heavy metals indicating a reduced bioavailability of these elements. This is particularly important for a safe utilization in agricultural soils to reduce the possible transfer of heavy metals to the food chain.

Antimicrobial Potentials of Flavonoids Isolated from Tagetes erecta

In this study, we are interested in a species of the family of Asteraceae (Tagetes erecta). This family is considered as a source of antimicrobial extracts with strong capacity. The extraction of the flavonoids is carried out by the method of liquid/liquid with the use of successive solvents. Afterwards, we evaluated the biological activity of the flavonoids on five pathogenic bacterial stocks such as Escherichia coli, Bacillus subtilis, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus and two stocks of yeasts to knowing Candida albicans) and Saccharomyces cerevisiae, by employing the method of the aromatogramme starting from a solid disc. The result of the antimicrobial activity shows an action and a variable degree of sensitivity according to bacterial stocks tested. It will be noted that the flavonoids have an inhibiting effect on E. coli, B. subtilis, K. pneumoniae and S. aureus. But a resistance with respect to the extract by P. aeruginosa, C. albicans and S. cerevisiae is to be mentioned.

Bacteria Flora in the Gut and Respiratory Organs of Clarias gariepinus in Fresh and Brackish Water Habitats of Ondo State, South/West Nigeria

Bacteria flora of Clarias gariepinus collected from two natural habitats namely Owena River (freshwater) and Igbokoda lagoon (brackish water) were examined using standard microbiological procedures. Thirteen bacterial species were identified. The result indicated that from the identified bacteria isolated, Vibrio sp, Proteus sp. Shigella sp. and E. coli were present in both habitats (fresh and brackish waters). Others were habitat-selective such as Salmonella sp., Pseudomonas sp, Enterococcus sp, Staphylococcus sp. that were found only in freshwater habitat. While Branhamella sp, Streptococcus sp. and Micrococcus sp. were found in brackish water habitat. Bacteria load from Owena river (freshwater) was found to be the highest load recorded at 6.21 x 104cfu. T-test analysis also revealed that there was a marked significant difference between bacterial load in guts of sampled Clarias from fresh water and brackish water habitats.

SFE as a Superior Technique for Extraction of Eugenol-Rich Fraction from Cinnamomum tamala Nees (Bay Leaf) - Process Analysis and Phytochemical Characterization

Highest yield of eugenol-rich fractions from Cinnamomum tamala (bay leaf) leaves were obtained by supercritical carbon dioxide (SC-CO2), compared to hydro-distillation, organic solvents, liquid CO2 and subcritical CO2 extractions. Optimization of SC-CO2 extraction parameters was carried out to obtain an extract with maximum eugenol content. This was achieved using a sample size of 10g at 55°C, 512 bar after 60min at a flow rate of 25.0 cm3/sof gaseous CO2. This extract has the best combination of phytochemical properties such as phenolic content (1.77mg gallic acid/g dry bay leaf), reducing power (0.80mg BHT/g dry bay leaf), antioxidant activity (IC50 of 0.20mg/ml) and anti-inflammatory potency (IC50 of 1.89mg/ml). Identification of compounds in this extract was performed by GC-MS analysis and its antimicrobial potency was also evaluated. The MIC values against E. coli, P. aeruginosa and S. aureus were 0.5, 0.25 and 0.5mg/ml, respectively. 

Cloning and Functional Characterization of Promoter Elements of the D Hordein Gene from the Barley (Hordeum vulgare L.) by Bioinformatic Tools

The low level of foreign genes expression in transgenic plants is a key factor that limits plant genetic engineering. Because of the critical regulatory activity of the promoters on gene transcription, they are studied extensively to improve the efficiency of the plant transgenic system. The strong constitutive promoters, such as CaMV 35S promoter and Ubiqutin 1 maize are usually used in plant biotechnology research. However the expression level of the foreign genes in all tissues is often undesirable. But using a strong seed-specific promoter to limit gene expression in the seed solves such problems. The purpose of this study is to isolate one of the seed specific promoters of Hordeum vulgare. So one of the common varieties of Hordeum vulgare in Iran was selected and their genomes extracted then the D-Hordein promoter amplified using the specific designed primers. Then the amplified fragment of the insert cloned in an appropriate vector and then transformed to E. coli. At last for the final admission of accuracy the cloned fragments sent for sequencing. Sequencing analysis showed that the cloned fragment DHPcontained motifs; like TATA box, CAAT-box, CCGTCC-box, AMYBOX1 and E-box etc., which constituted the seed-specific promoter activity. The results were compared with sequences existing in data banks. D-Hordein promoters of Alger has 99% similarity at 100 % coverage. The results also showed that D-Hordein promoter of barley and HMW promoter of wheat are too similar.

Antimicrobial Activity of Girardinia heterophylla

In the present study an attempt has been made to prepare the crude extracts of leaves and stem of ‘Girardinia heterophylla’ by using various solvents like petroleum ether, ethanol and double distilled water. The samples were given the code NGLS 1, NGLS 2, NGLS 3 and NGSS 1, NGSS 2 and NGSS 3 respectively. All the extracts were used to study their antimicrobial activity against gram positive bacteria e.g. Bacillus subtilis, gram negative bacteria e.g. E. coli and K. pneumonia and antifungal activity against Aspergillus niger. The results of the antimicrobial activity showed that all the crude extracts of the plant possesses antibacterial activity. Maximum antibacterial activity was shown by NGLS 2, NGLS 3 and NGSS 3 against K. pneumonia. The growth of fungus A. niger was also inhibited by all the crude extracts. Maximum inhibition was shown by NGSS 2 followed by NGSS 1.

Statistical Optimization of Process Conditions for Disinfection of Water Using Defatted Moringa oleifera Seed Extract

In this study, statistical optimization design was used to study the optimum disinfection parameters using defatted crude Moringa oleifera seed extracts against Escherichia coli (E. coli) bacterial cells. The classical one-factor-at-a-time (OFAT) and response surface methodology (RSM) was used. The possible optimum range of dosage, contact time and mixing rate from the OFAT study were 25mg/l to 200mg/l, 30minutes to 240 minutes and 100rpm to 160rpm respectively. Analysis of variance (ANOVA) of the statistical optimization using faced centered central composite design showed that dosage, contact time and mixing rate were highly significant. The optimum disinfection range was 125mg/l, at contact time of 30 minutes with mixing rate of 120 rpm. 

High Efficiency, Selectivity against Cancer Cell Line of Purified L-Asparaginase from Pathogenic Escherichia coli

L-asparaginase was extracted from pathogenic Escherichia coli which was isolated from urinary tract infection patients. L-asparaginase was purified 96-fold by ultrafiltration, ion exchange and gel filtration giving 39.19% yield with final specific activity of 178.57 IU/mg. L-asparaginase showed 138,356±1,000 Dalton molecular weight with 31024±100 Dalton molecular mass. Kinetic properties of enzyme resulting 1.25×10-5 mM Km and 2.5×10-3 M/min Vmax. L-asparaginase showed a maximum activity at pH 7.5 when incubated at 37 ºC for 30 min and illustrated its full activity (100%) after 15 min incubation at 20-37 ºC, while 70% of its activity was lost when incubated at 60 ºC. L-asparaginase showed cytotoxicity to U937 cell line with IC50 0.5±0.19 IU/ml, and selectivity index (SI=7.6) about 8 time higher selectivity over the lymphocyte cells. Therefore, the local pathogenic E. coli strains may be used as a source of high yield of L-asparaginase to produce anti cancer agent with high selectivity.

Study the Biological Activities of Tribulus Terrestris Extracts

In this study the extracts of the Iraqi herb Tribulus terrestris (Al-Hassage or Al-Kutub) was done by using of polar and non polar solvents, then the biological activity of these extractants was studied in three fields, First, the antibacterial activity (in vitro) on gram positive bacteria (Staphylococcus aureus), and gram negative bacteria (E. coli, Proteus vulgaris, Pseudomonas aerugiuosa, and Klebsiella), all extracts showed considerable activity against all bacteria. Second, the effect of extracts on free serum testosterone level in male mice (in vivo), the alcoholic, and acetonitrilic extracts showed significant (P < 0.05) increase in free serum testosterone level, and we found that the extracts contained compounds with less genotoxic effects in mice germ cells. 3rd, was to study the effect of methanolic extract of T. terrestris in diabetes management.

Placebo Effect on Psychoemotional Status and Clinical Analyses of Armenian Crohn's Disease Patients

The Zung self-depression scale and Beck Anxiety Inventory were used both to study the depression and anxiety levels of Armenian Crohn’s disease patients, and to reveal the relation between emotional status and placebo effect of these patients. On the other hand, the blood cell analyses and gut bacteria investigations were used to assess the placebo effect on ESR, and haemoglobin-s and leukocyte-s levels as well as gut commensal E. coli quantities of these patients. Despite of registered high levels of depression and anxiety, the high placebo effect on psychoemotional status for investigated patients during the investigations was described. On the other hand, no positive effect of placebo on measurements of ESP and hemoglobin-s levels of Crohn’s disease patients was revealed. The importance of use of psychotherapies for optimal outcomes during treatments of Crohn’s disease is discussed.

Synthesis, Investigation, DFT Study and Biological Activity of Zirconium (IV) Complexes

Zirconium diamine and triamine complexes can possess biological activities. These complexes were synthesised via the reaction of equimolar quantities of 1,10-phenanthroline {NC3H3(C6H2)NC3H3} (L1) or 4-4-amino phenazone {ONC6H5(NH)CH(NH2} (L2) or diphenyl carbizon {HNNCO(NH)2(C6H5)} (L3) with a Zirconium Salt {ZrOCl2} in a 1:1 ratio to form complexes [{NC3H3(C6H2)NC3H3}ZrOCl2}] [ZrOCl2L1], [{(O2NC6H4(NH)(NH2)}ZrOCl2] [ZrOCl2L2] and [{HNNCO(NH)2(C6H5)ZrOCl2}] [ZrOCl2L3] respectively. They were characterised using Fourier Transform Infrared (FT-IR) and UV-Visible spectroscopy. Also a variable temperature study of these complexes was completed, using UV-Visible spectroscopy to observe electronic transitions under temperature control. Also a DFT study was done on these complexes via the information from FT-IR and UV-Visible spectroscopy. These complexes were found to show different inhibition to the growth of bacterial strains of Bacillus spp. & Klebsiella spp. & E. coli & Proteus spp. & Pseudomona spp. at different concentrations (0.001, 0.2 and 1M). For better understanding these complexes were examined by using a Density Functional Theory (DFT) calculation.

Computational Identification of Bacterial Communities

Stable bacterial polymorphism on a single limiting resource may appear if between the evolved strains metabolic interactions take place that allow the exchange of essential nutrients [8]. Towards an attempt to predict the possible outcome of longrunning evolution experiments, a network based on the metabolic capabilities of homogeneous populations of every single gene knockout strain (nodes) of the bacterium E. coli is reconstructed. Potential metabolic interactions (edges) are allowed only between strains of different metabolic capabilities. Bacterial communities are determined by finding cliques in this network. Growth of the emerged hypothetical bacterial communities is simulated by extending the metabolic flux balance analysis model of Varma et al [2] to embody heterogeneous cell population growth in a mutual environment. Results from aerobic growth on 10 different carbon sources are presented. The upper bounds of the diversity that can emerge from single-cloned populations of E. coli such as the number of strains that appears to metabolically differ from most strains (highly connected nodes), the maximum clique size as well as the number of all the possible communities are determined. Certain single gene deletions are identified to consistently participate in our hypothetical bacterial communities under most environmental conditions implying a pattern of growth-condition- invariant strains with similar metabolic effects. Moreover, evaluation of all the hypothetical bacterial communities under growth on pyruvate reveals heterogeneous populations that can exhibit superior growth performance when compared to the performance of the homogeneous wild-type population.

A Novel Cytokine Derived Fusion Tag for Over- Expression of Heterologous Proteins in E. coli

We report a novel fusion tag for expressing recombinant proteins in E. coli. The fusion tag is the C-terminus part of the human GMCSF gene comprising 45 amino acids, which aid in over expression of otherwise non expressible genes. Expression of hIFN a2b with this fusion tag also escapes the requirement of rare codons for expression. This is also a first report of a small fusion tag of human origin having affinity to heparin sepharose column facilitating the purification of fusion protein.

The Effect of Sodium Chloride and pH on the Antimicrobial Effectiveness of Essential Oils Against Pathogenic and Food Spoilage Bacteria:Implications in Food Safety

The purpose of this study was to elucidate the factors affecting antimicrobial effectiveness of essential oils against food spoilage and pathogenic bacteria. The minimum inhibition concentrations (MIC) of the essential oils, were determined by turbidimetric technique using Biocreen C, analyzer. The effects of pH ranging from 7.3 to 5.5 in absence and presence of essential oils and/or NaCl on the lag time and mean generation time of the bacteria at 370C, were carried out and results were determined showed that, combination of low pH and essential oil at 370C had additive effects against the test micro-organisms. The combination of 1.2 % (w/v) of NaCl and clove essential oil at 0.0325% (v/v) was effective against E. coli. The use of concentrations less than MIC in combination with low pH and or NaCl has the potential of being used as an alternative to “traditional food preservatives".

Prevalence and Antimicrobial Susceptibility Patterns of Enteric Bacteria Isolated from Water and Fish in Lake Victoria Basin of Western Kenya

A cross sectional study design and standard microbiological procedures were used to determine the prevalence and antimicrobial susceptibility patterns of Escherichia coli, Salmonella enterica serovar typhimurium and Vibrio cholerae O1 isolated from water and two fish species Rastrineobola argentea and Oreochromis niloticus collected from fish landing beaches and markets in the Lake Victoria Basin of western Kenya. Out of 162 samples analyzed, 133 (82.1%) were contaminated, with S. typhimurium as the most prevalent (49.6%), followed by E. coli (46.6%), and lastly V. cholerae (2.8%). All the bacteria isolates were sensitive to ciprofloxacin. E. coli isolates were resistant to ampicillin, tetracycline, cotrimoxazole, chloramphenical and gentamicin while S. typhimurium isolates exhibited resistance to ampicillin, tetracycline, and cotrimoxazole. The V. cholerae O1 isolates were resistant to tetracycline and ampicillin. The high prevalence of drug resistant enteric bacteria in water and fish from the study region needs public health intervention from the local government.

The Presence of Enterobacters (E.Coli and Salmonella spp.) in Industrial Growing Poultry in Albania

The development of the poultry industry in Albania is mainly based on the existence of intensive modern farms with huge capacities, which often are mixed with other forms. Colibacillosis is commonly displayed regardless of the type of breeding, delivering high mortality in poultry industry. The mechanisms with which pathogen enterobacters are able to cause the infection in poultry are not yet clear. The routine diagnose in the field, followed by isolation of E. coli and species of Salmonella genres in reference laboratories cannot lead in classification or full recognition of circulative strains in a territory, if it is not performed a differentiation among the present microorganisms in intensive farms and those in rural areas. In this study were isolated 1.496 strains of E. coli and 378 Salmonella spp. This study, presents distribution of poultry pathogenosity of E.coli and Salmonella spp., based on the usage of innovative diagnostic methods.

Some Peculiarities of Growth and Functional Activity of Escherichia coli Strain from Probiotic Formula “ASAP“

It has been shown that pH 7,3 and 37 0C are the optimal condition for the growth of E. coli “ASAP". The cells grow well on Glucose, Lactose, D-Mannitol, D-Sorbitol, (+)-Xylose, L- (+)-Arabinose and Dulcitol. No growth has been observed on Sucrose, Inositol, Phenylalanine, and Tryptophan. The strain is sensitive to a range of antibiotics. The present study has demonstrated that E. coli “ASAP" inhibit the growth of S. enterica ATCC #700931 in vitro. The studies on conjugating activity has revealed no conjugant of E. coli “ASAP" with plasmid strains E. coli G35#59 and S. enterica ATCC #700931. On the other hand, the conjugants with low frequencies were obtained from E. coli “ASAP" with E. coli G35#61, and E. coli “ASAP" with randomly chosen isolate from healthy human gut microflora: E. coli E6. The results of present study have demonstrated improvements in gut microflora condition of patients with different diseases after the administration of “ASAP"

Antimicrobial, Antiplasmid and Cytotoxicity Potentials of Marine Algae Halimeda opuntia and Sarconema filiforme Collected from Red Sea Coast

The antimicrobial, antiplasmid and cytotoxic activities of marine algae Halimeda opuntia and Sarconema filiforme were investigated. Antimicrobial bioassay against some human pathogenic bacteria and yeast were conducted using disc diffusion method. Halimeda extract exhibited antibacterial activity against six species of microrganisms, with significant inhibition against Staphylococcus aureus. While Sarconema extract was better potent as antifungal against Candida albicans. Comparative antibacterial studies showed that Halimeda extract showed equivalent or better activity as compared with commercial antibiotic when tested against Staphylococcus aureus. Further tests conducted using dilution method showed both extracts as having bacteriostatic mode of action against the tested microorganisms. Methanol extract of two species showed significant cytotoxicity (LC50

Detection of Pathogenic Escherichia coli Strains Pollution in Red Deer Meat in Latvia and Determination the Compatibility of VT1, VT2, eae A Genes in their Isolate

Tasks of the work were study the possible E.coli contamination in red deer meat, identify pathogenic strains from isolated E.coli, determine their incidence in red deer meat and determine the presence of VT1, VT2 and eaeA genes for the pathogenic E.coli. 8 (10%) samples were randomly selected from 80 analysed isolates of E.coli and PCR reaction was performed on them. PCR was done both on initial materials – samples of red deer meat - and for already isolated liqueurs. Two of analysed venison samples contain verotoxin-producing strains of E. coli. It means that this meat is not safe to consumer. It was proven by the sequestration reaction of E. coli and by comparison of the obtained results with the database of microorganism genome available on the internet that the isolated culture corresponds to region 16S rDNS of E. coli thus presenting correctness of the microbiological methods.

Construction of Recombinant E.coli Expressing Fusion Protein to Produce 1,3-Propanediol

In this study, a synthetic pathway was created by assembling genes from Clostridium butyricum and Escherichia coli in different combinations. Among the genes were dhaB1 and dhaB2 from C. butyricum VPI1718 coding for glycerol dehydratase (GDHt) and its activator (GDHtAc), respectively, involved in the conversion of glycerol to 3-hydroxypropionaldehyde (3-HPA). The yqhD gene from E.coli BL21 was also included which codes for an NADPHdependent 1,3-propanediol oxidoreductase isoenzyme (PDORI) reducing 3-HPA to 1,3-propanediol (1,3-PD). Molecular modeling analysis indicated that the conformation of fusion protein of YQHD and DHAB1 was favorable for direct molecular channeling of the intermediate 3-HPA. According to the simulation results, the yqhD and dhaB1 gene were assembled in the upstream of dhaB2 to express a fusion protein, yielding the recombinant strain E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP41Y3). Strain BP41Y3 gave 10-fold higher 1,3-PD concentration than E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP31Y2) expressing the recombinant enzymes simultaneously but in a non-fusion mode. This is the first report using a gene fusion approach to enhance the biological conversion of glycerol to the value added compound 1,3- PD.