Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 111

Search results for: E.coli

111 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

Authors: S. Liepina, A. Jemeljanovs

Abstract:

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.

Keywords: Deer meat, pathogenic Escherichia coli

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

Authors: Marine A. Balayan, Susanna S. Mirzabekyan, Marine Isajanyan, Zaven S. Pepoyan, Аrmen H. Trchounian, Аstghik Z. Pepoyan, Helena Bujdakova

Abstract:

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"

Keywords: E. coli, ASAP, Probiotic formula, gut microflora.

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

Authors: Boci J., Çabeli P., Shtylla T., Kumbe I.

Abstract:

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.

Keywords: poultry, E.coli, Salmonella spp., Enterobacter

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108 Characterization of Silica Nanoparticles in Interaction with Escherichia coli Bacteria

Authors: Ibtissem Gammoudi, Ndeye Rokhaya Faye, Fabien Moroté, Daniel Moynet, Christine Grauby-Heywang, Touria Cohen-Bouhacina

Abstract:

The objective of the present investigation was to evaluate the morphology of Escherchia coli bacteria in interaction with SiO2 nanoparticles. This study was made by atomic force microscopy and quartz crystal microbalance using SiO2 nanoparticles with 10nm, 50nm and 100nm diameter and bacteria immobilized on polyelectrolyte multilayer films obtained by spin coating or by “layer by layer” (LbL) method.

Keywords: Atomic Force Microscopy, Escherichia coli, Quartz Crystal Microbalance, polyelectrolyte, silica nanoparticle.

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107 Antibacterial Capacity of Plumeria alba Petals

Authors: M. H. Syakira, L. Brenda

Abstract:

Antibacterial activity of Plumeria alba (Frangipani) petals methanolic extracts were evaluated against Escherichia coli, Proteus vulgaris,Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus saprophyticus, Enterococcus faecalis and Serratia marcescens by using disk diffusion method. Concentration extracts (80 %) showed the highest inhibition zone towards Escherichia coli (14.3 mm). Frangipani extract also showed high antibacterial activity against Staphylococcus saprophyticus, Proteus vulgaris and Serratia marcescens, but not more than the zones of the positive control used. Comparison between two broad specrum antibiotics to frangipani extracts showed that the 80 % concentration extracts produce the same zone of inhibition as Streptomycin. Frangipani extracts showed no bacterial activity towards Klebsiella pneumoniae, Pseudomonas aeruginosa and Enterococcus faecalis. There are differences in the sensitivity of different bacteria to frangipani extracts, suggesting that frangipani-s potency varies between these bacteria. The present results indicate that frangipani showed significant antibacterial activity especially to Escherichia coli.

Keywords: Frangipani, Plumeria alba, anti microbial, Escherichia coli

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

Authors: Rosarin Rujananon, Poonsuk Prasertsan, Amornrat Phongdara, Tanate Panrat, Jibin Sun, Sugima Rappert, An-Ping Zeng

Abstract:

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.

Keywords: Recombinant E.coli, 1, 3-propanediol, glycerol, fusion protein.

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105 Clinical Signs of Neonatal Calves in Experimental Colisepticemia

Authors: Samad Lotfollahzadeh

Abstract:

Escherichia coli (E. coli) is the most isolated bacteria from blood circulation of septicemic calves. Given the prevalence of septicemia in animals and its economic importance in veterinary practice, better understanding of changes in clinical signs following disease, may contribute to early detection of disorder. The present study has been carried out to detect changes of clinical signs in induced sepsis in calves with E. coli. Colisepticemia has been induced in 10 twenty-day old healthy Holstein- Frisian calves with intravenous injection of 1.5 X 109 colony forming units (cfu) of O111:H8 strain of E. coli. Clinical signs including rectal temperature, heart rate, respiratory rate, shock, appetite, sucking reflex, feces consistency, general behavior, dehydration and standing ability were recorded in experimental calves during 24 hours after induction of colisepticemia. Blood culture was also carried out from calves four times during experiment. ANOVA with repeated measure is used to see changes of calves’ clinical signs to experimental colisepticemia, and values of P≤ 0.05 was considered statistically significant. Mean values of rectal temperature and heart rate as well as median values of respiratory rate, appetite, suckling reflex, standing ability and feces consistency of experimental calves increased significantly during study (P<0.05). In the present study median value of shock score was not significantly increased in experimental calves (P> 0.05). The results of present study showed that total score of clinical signs in calves with experimental colisepticemia increased significantly, although score of some clinical signs such as shock did not change significantly.

Keywords: Calves, Clinical signs scoring, E. coli O111:H8, Experimental colisepticemia,

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104 An Alternative Antimicrobial Approach to Fight Bacterial Pathogens from Phellinus linteus

Authors: S. Techaoei, K. Jarmkom, P. Eakwaropas, W. Khobjai

Abstract:

The objective of this research was focused on investigating in vitro antimicrobial activity of Phellinus linteus fruiting body extracts on Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Phellinus linteus fruiting body was extracted with ethanol and ethyl acetate and was vaporized. The disc diffusion assay was used to assess antimicrobial activity against tested bacterial strains. Primary screening of chemical profile of crude extract was determined by using thin layer chromatography. The positive control and the negative control were used as erythromycin and dimethyl sulfoxide, respectively. Initial screening of Phellinus linteus crude extract with the disc diffusion assay demonstrated that only ethanol had greater antimicrobial activity against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. The MIC assay showed that the lower MIC was observed with 0.5 mg/ml of Pseudomonas aeruginosa and Methicillin-resistant Staphylococcus aureus and 0.25 mg/ml. of Escherichia coli and Staphylococcus aureus, respectively. TLC chemical profile of extract was represented at Rf ≈ 0.71-0.76.

Keywords: Staphylococcus aureus, Phellinus linteus, methicillin-resistant Staphylococcus aureus, antimicrobial activity, Escherichia coli.

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

Authors: S. Banerjee, A. Apte Deshpande, N. Mandi, S. Padmanabhan

Abstract:

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.

Keywords: fusion tag, bacterial expression, rare codons, human GMCSF

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

Authors: Hazim Saadoon Aljewari, Mohammed Ibraheem Nader, Abdul Hussain M. Alfaisal, NatthidaWeerapreeyakul, Sahapat

Abstract:

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.

Keywords: L-asparaginase, Purification, Cytotoxicity, selectivity index

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101 Biosynthesis of Titanium Dioxide Nanoparticles and Their Antibacterial Property

Authors: Prachi Singh

Abstract:

This paper presents a low-cost, eco-friendly and reproducible microbe mediated biosynthesis of TiO2 nanoparticles. TiO2 nanoparticles synthesized using the bacterium, Bacillus subtilis, from titanium as a precursor, were confirmed by TEM analysis. The morphological characteristics state spherical shape, with the size of individual or aggregate nanoparticles, around 30-40 nm. Microbial resistance represents a challenge for the scientific community to develop new bioactive compounds. Here, the antibacterial effect of TiO2 nanoparticles on Escherichia coli was investigated, which was confirmed by CFU (Colony-forming unit). Further, growth curve study of E. coli Hb101 in the presence and absence of TiO2 nanoparticles was done. Optical density decrease was observed with the increase in the concentration of TiO2. It could be attributed to the inactivation of cellular enzymes and DNA by binding to electron-donating groups such as carboxylates, amides, indoles, hydroxyls, thiols, etc. which cause little pores in bacterial cell walls, leading to increased permeability and cell death. This justifies that TiO2 nanoparticles have efficient antibacterial effect and have potential to be used as an antibacterial agent for different purposes.

Keywords: Antibacterial effect, CFU, Escherichia coli Hb101, growth curve, TEM, TiO2 nanoparticle, toxicity, UV-Vis.

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

Authors: Suleyman A. Muyibi, Munirat, A. Idris, Saedi Jami, Parveen Jamal, Mohd Ismail Abdul Karim

Abstract:

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. 

Keywords: E.coli, disinfection, Moringa oleifera, response surface methodology.

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99 Emergence of Fluoroquinolone Resistance in Pigs, Nigeria

Authors: Igbakura I. Luga, Alex A. Adikwu

Abstract:

A comparison of resistance to quinolones was carried out on isolates of Shiga toxin-producing Escherichia coliO157:H7 from cattle and mecA and nuc genes harbouring Staphylococcus aureus from pigs. The isolates were separately tested in the first and current decades of the 21st century. The objective was to demonstrate the dissemination of resistance to this frontline class of antibiotic by bacteria from food animals and bring to the limelight the spread of antibiotic resistance in Nigeria. A total of 10 isolates of the E. coli O157:H7 and 9 of mecA and nuc genes harbouring S. aureus were obtained following isolation, biochemical testing, and serological identification using the Remel Wellcolex E. coli O157:H7 test. Shiga toxin-production screening in the E. coli O157:H7 using the verotoxin E. coli reverse passive latex agglutination (VTEC-RPLA) test; and molecular identification of the mecA and nuc genes in S. aureus. Detection of the mecA and nuc genes were carried out using the protocol by the Danish Technical University (DTU) using the following primers mecA-1:5'-GGGATCATAGCGTCATTATTC-3', mecA-2: 5'-AACGATTGTGACACGATAGCC-3', nuc-1: 5'-TCAGCAAATGCATCACAAACAG-3', nuc-2: 5'-CGTAAATGCACTTGCTTCAGG-3' for the mecA and nuc genes, respectively. The nuc genes confirm the S. aureus isolates and the mecA genes as being methicillin-resistant and so pathogenic to man. The fluoroquinolones used in the antibiotic resistance testing were norfloxacin (10 µg) and ciprofloxacin (5 µg) in the E. coli O157:H7 isolates and ciprofloxacin (5 µg) in the S. aureus isolates. Susceptibility was tested using the disk diffusion method on Muller-Hinton agar. Fluoroquinolone resistance was not detected from isolates of E. coli O157:H7 from cattle. However, 44% (4/9) of the S. aureus were resistant to ciprofloxacin. Resistance of up to 44% in isolates of mecA and nuc genes harbouring S. aureus is a compelling evidence for the rapid spread of antibiotic resistance from bacteria in food animals from Nigeria. Ciprofloxacin is the drug of choice for the treatment of Typhoid fever, therefore widespread resistance to it in pathogenic bacteria is of great public health significance. The study concludes that antibiotic resistance in bacteria from food animals is on the increase in Nigeria. The National Food and Drug Administration and Control (NAFDAC) agency in Nigeria should implement the World Health Organization (WHO) global action plan on antimicrobial resistance. A good starting point can be coordinating the WHO, Office of International Epizootics (OIE), Food and Agricultural Organization (FAO) tripartite draft antimicrobial resistance monitoring and evaluation (M&E) framework in Nigeria.

Keywords: Fluoroquinolone, Nigeria, resistance, Staphylococcus aureus.

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98 Modified Genome-Scale Metabolic Model of Escherichia coli by Adding Hyaluronic Acid Biosynthesis-Related Enzymes (GLMU2 and HYAD) from Pasteurella multocida

Authors: P. Pasomboon, P. Chumnanpuen, T. E-kobon

Abstract:

Hyaluronic acid (HA) consists of linear heteropolysaccharides repeat of D-glucuronic acid and N-acetyl-D-glucosamine. HA has various useful properties to maintain skin elasticity and moisture, reduce inflammation, and lubricate the movement of various body parts without causing immunogenic allergy. HA can be found in several animal tissues as well as in the capsule component of some bacteria including Pasteurella multocida. This study aimed to modify a genome-scale metabolic model of Escherichia coli using computational simulation and flux analysis methods to predict HA productivity under different carbon sources and nitrogen supplement by the addition of two enzymes (GLMU2 and HYAD) from P. multocida to improve the HA production under the specified amount of carbon sources and nitrogen supplements. Result revealed that threonine and aspartate supplement raised the HA production by 12.186%. Our analyses proposed the genome-scale metabolic model is useful for improving the HA production and narrows the number of conditions to be tested further.

Keywords: Pasteurella multocida, Escherichia coli, hyaluronic acid, genome-scale metabolic model, bioinformatics.

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97 Surface Charge Based Rapid Method for Detection of Microbial Contamination in Drinking Water and Food Products

Authors: Kandpal M. , Gundampati R. K , Debnath M.

Abstract:

Microbial contamination, most of which are fecal born in drinking water and food industry is a serious threat to humans. Escherichia coli is one of the most common and prevalent among them. We have developed a sensor for rapid and an early detection of contaminants, taking E.coli as a threat indicator organism. The sensor is based on co-polymerizations of aniline and formaldehyde in form of thin film over glass surface using the vacuum deposition technique. The particular doping combination of thin film with Fe-Al and Fe-Cu in different concentrations changes its non conducting properties to p- type semi conductor. This property is exploited to detect the different contaminants, believed to have the different surface charge. It was found through experiments that different microbes at same OD (0.600 at 600 nm) have different conductivity in solution. Also the doping concentration is found to be specific for attracting microbes on the basis of surface charge. This is a simple, cost effective and quick detection method which not only decreases the measurement time but also gives early warnings for highly contaminated samples.

Keywords: Sensor, Vacuum deposition technique, thin film, E.coli detection, doping concentration.

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96 Novel Inhibitor of E. coli DNA Adenine Methyltransferase (Ecodam)

Authors: H. Elsawy, A. Jeltsch

Abstract:

EcoDam is an adenine-N6 DNA methyltransferase that methylates the GATC sites in the Escherichia coli genome. DNA-adenine methylation is not present in higher eukaryotes including humans. These observations raise the possibility that dam inhibitors may be used as anti-microbial agents. Polyphosphate (Poly(P)) is an important metabolite and signaling molecule in prokaryotes and eukaryotes. Here, by using gel retardation experiments to investigate the competition of DNA binding by EcoDam in the presence of polyphosphate, we found that Poly (P) strongly interferes with DNA binding by EcoDam, while same concentration of monophosphate does not. In addition, we demonstrated that Poly (P) binding inhibits the activity of EcoDam and our results suggest that Poly (P) led to strong inhibition of the EcoDam catalytic activity, while monophosphate had only moderate effect.

Keywords: Antibacterial drugs, EcoDam inhibitors, Polyphosphate.

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95 X-ray Crystallographic Analysis of MinC N-Terminal Domain from Escherichia coli

Authors: Jun Yop An, Kyoung Ryoung Park, Jung-Gyu Lee, Hyung-Seop Youn, Jung-Yeon Kang, Gil Bu Kang, Soo Hyun Eom

Abstract:

MinC plays an important role in bacterial cell division system by inhibiting FtsZ assembly. However, the molecular mechanism of the action is poorly understood. E. coli MinC Nterminus domain was purified and crystallized using 1.4 M sodium citrate pH 6.5 as a precipitant. X-ray diffraction data was collected and processed to 2.3 Å from a native crystal. The crystal belonged to space group P212121, with the unit cell parameters a = 52.7, b = 54.0, c = 64.7 Å. Assuming the presence of two molecules in the asymmetric unit, the Matthews coefficient value is 1.94 Å3 Da-1, which corresponds to a solvent content of 36.5%. The overall structure of MinCN is observed as a dimer form through anti-parallel ß-strand interaction.

Keywords: MinC, Cell division, Crystallization.

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

Authors: Jackson H. O. Onyuka, Rose Kakai, David M. Onyango, Peter F. Arama, John Gichuki, Ayub V.O. Ofulla

Abstract:

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.

Keywords: Aquatic environments, Antimicrobial resistance, Enteric bacteria, Lake Victoria Basin

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

Authors: G. Petruzzelli, F. Pedron, M. Grifoni, A. Pera, I. Rosellini, B. Pezzarossa

Abstract:

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.

Keywords: Biological sludge, Ca(OH)2, copper, pathogens, sanitation, zinc.

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92 Incidence of Pathogenic Bacteria in Cakes and Tarts Displayed for Sale in Tripoli, Libya

Authors: Sabrin M. Al-Jafaeri, Nuri S. Madi, Mohamed H. Nahaisi

Abstract:

This study was conducted to investigate the incidence of pathogenic bacteria: Salmonella, Shigella, Escherichia coli O157 and Staphylococcus aureus in cakes and tarts collected from thirtyfive confectionery producing and selling premises located within Tripoli city, Libya. The results revealed an incidence of S. aureus with 94.4 and 48.0 %, E. coli O157 with 14.7 and 4.0 % and Salmonella sp. with 5.9 and 8.0 % in cakes and tarts samples respectively; while Shigella was not detected in all samples. In order to determine the source of these pathogenic bacteria, cotton swabs were taken from the hands of workers on the production line, the surfaces of preparation tables and cream whipping instruments. The results showed that the cotton swabs obtained from the hands of workers contained S. aureus and Salmonella sp. with an incidence of 42.9 and 2.9 %, the cotton swabs obtained from the surfaces of preparation tables 22.9 and 2.9 % and the cotton swabs obtained from the cream whipping instruments 14.3 and 0.0 % respectively; while E. coli O157 and Shigella sp. were not detected in all swabs. Additionally, other bacteria were isolated from the hands of workers and the Surfaces of producing equipments included: Aeromonas sp., Pseudomonas sp., E. coli, Klebsiella sp., Enterobacter sp., Citrobacter sp., Proteus sp., Serratia sp. and Acinetobacter sp. These results indicate that some of the cakes and tarts might pose threat to consumer's health. Meanwhile, occurrences of pathogenic bacteria on the hands of those who are working in production line and the surfaces of equipments reflect poor hygienic practices at most confectionery premises examined in this study. Thus, firm and continuous surveillance of these premises is needed to insure the consumer's health and safety.

Keywords: Pathogenic bacteria, Cakes, Tarts, Confectionery producing and selling premises.

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91 Assessing the Antimicrobial Activity of Chitosan Nanoparticles by Fluorescence-Labeling

Authors: Laidson P. Gomes, Cristina T. Andrade, Eduardo M. Del Aguila, Cameron Alexander, Vânia M. F. Paschoalin

Abstract:

Chitosan is a natural polysaccharide prepared by the N-deacetylation of chitin. In this study, the physicochemical and antibacterial properties of chitosan nanoparticles, produced by ultrasound irradiation, were evaluated. The physicochemical properties of the nanoparticles were determined by dynamic light scattering and zeta potential analysis. Chitosan nanoparticles inhibited the growth of E. coli. The minimum inhibitory concentration (MIC) values were lower than 0.5 mg/mL, and the minimum bactericidal concentration (MBC) values were similar or higher than MIC values. Confocal laser scanning micrographs (CLSM) were used to observe the interaction between E. coli suspensions mixed with FITC-labeled chitosan polymers and nanoparticles.

Keywords: Chitosan nanoparticles, dynamic light scattering, zeta potential, confocal microscopy, antibacterial activity.

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90 Study of Microbial Critical Points of Saffron from Farm to Factory in Iran

Authors: N. Khazaei, M. Jouki, A. Kalbasi, H. Tavakolipour, S. Rajabifar, F. Motamedi. Sedeh, A. Jouki

Abstract:

In this research saffron samples were prepared from farms and sampling was done in four states contain : sampling from fresh saffron of petal with forceps , sampling from fresh saffron of petal by hands, sampling from dried sample by warm air in shadow, sampling from dried sample which dried by dryer. Samples collected and kept in sterile tubes and containers and carried to laboratory and maintained until experiment. Microbial experiments were performed to determine microbial load such as total count, Staphylococcus aureus, coli form, E.coli, mold and yeast. Results showed that in picking and drying stages the contamination amount increases in saffron samples. There was a significant difference between the microbial load of picked up saffron by forceps and by hands, and also between dried saffron by warm air in shadow and by dryer.

Keywords: saffron; contaminations; preparation method

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89 Paper-Based Colorimetric Sensor Utilizing Peroxidase-Mimicking Magnetic Nanoparticles Conjugated with Aptamers

Authors: Min-Ah Woo, Min-Cheol Lim, Hyun-Joo Chang, Sung-Wook Choi

Abstract:

We developed a paper-based colorimetric sensor utilizing magnetic nanoparticles conjugated with aptamers (MNP-Apts) against E. coli O157:H7. The MNP-Apts were applied to a test sample solution containing the target cells, and the solution was simply dropped onto PVDF (polyvinylidene difluoride) membrane. The membrane moves the sample radially to form the sample spots of different compounds as concentric rings, thus the MNP-Apts on the membrane enabled specific recognition of the target cells through a color ring generation by MNP-promoted colorimetric reaction of TMB (3,3',5,5'-tetramethylbenzidine) and H2O2. This method could be applied to rapidly and visually detect various bacterial pathogens in less than 1 h without cell culturing.

Keywords: Aptamer, colorimetric sensor, E. coli O157:H7, magnetic nanoparticle, polyvinylidene difluoride.

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88 Antimicrobial Potentials of Flavonoids Isolated from Tagetes erecta

Authors: N. Behidj-Benyounes, S. Bennaamane, F. Zohra Bissaad, N. Chebouti, H. Mohandkaci, N. Abdalaziz, S. Iddou

Abstract:

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.

Keywords: Antimicrobial activity, flavonoids, microbial strains, Tagetes erecta L.

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87 Preliminary Study of Antimicrobial Activity against Escherichia coli and Probiotic Properties of Lactic Acid Bacteria Isolated from Thailand Fermented Foods

Authors: Phanwipa Pangsri, Yawariyah Weahayee

Abstract:

The lactic acid bacteria (LAB) were isolated from 10 samples of fermented foods (Sa-tor-dong and Bodo) in South locality of Thailand. The 23 isolates of lactic acid bacteria were selected, which were exhibited a clear zone and growth on MRS agar supplemented with CaCO3. All of lactic acid bacteria were tested on morphological and biochemical. The result showed that all isolates were Gram’s positive, non-spore forming but only 10 isolates displayed catalase negative. The 10 isolates including BD1 .1, BD 1.2, BD 2.1, BD2.2, BD 2.3, BD 3.1, BD 4.1, BD 5.2, ST 4.1 and ST 5.2 were selected for inhibition activity determination. Only 2 strains (ST 4.1 and BD 2.3) showed inhibition zone on agar, when using Escherichia coli sp. as target strain. The ST 4.1 showed highest inhibition zone on agar, which was selected for probiotic property testing. The ST4.1 isolate could grow in MRS broth containing a high concentration of sodium chloride 6%, bile salts 7%, pH 4-10 and vary temperature at 15-45°C.

Keywords: Lactic acid bacteria, Probiotic, Antimicrobial.

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86 Synthesis, Characterization and Coating of the Zinc Oxide Nanoparticles on Cotton Fabric by Mechanical Thermo-Fixation Techniques to Impart Antimicrobial Activity

Authors: Imana Shahrin Tania, Mohammad Ali

Abstract:

The present study reports the synthesis, characterization and application of nano-sized zinc-oxide (ZnO) particles on a cotton fabric surface. The aim of the investigations is to impart the antimicrobial activity on textile cloth. Nanoparticle is synthesized by wet chemical method from zinc sulphate and sodium hydroxide. SEM (scanning electron micrograph) images are taken to demonstrate the surface morphology of nanoparticles. XRD analysis is done to determine the crystal size of the nanoparticle. With the conformation of nanoformation, the cotton woven fabric is treated with ZnO nanoparticle by mechanical thermo-fixation (pad-dry-cure) technique. To increase the wash durability of nano treated fabric, an acrylic binder is used as a fixing agent. The treated fabric shows up to 90% bacterial reduction for S. aureus (Staphylococcus aureus) and 87% for E. coli (Escherichia coli) which is appreciable for bacteria protective clothing.

Keywords: Nanoparticle, zinc oxide, cotton fabric, antibacterial activity, binder.

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85 Cloning and Expression of D-Threonine Aldolase from Ensifer arboris NBRC100383

Authors: Sang-Ho Baik

Abstract:

D-erythro-cyclohexylserine (D chiral unnatural β-hydroxy amino acid expected for the synthesis of drug for AIDS treatment. To develop a continuous bioconversion system with whole cell biocatalyst of D-threonine aldolase (D genes for the D-erythro-CHS production, D-threonine aldolase gene was amplified from Ensifer arboris 100383 by direct PCR amplication using two degenerated oligonucleotide primers designed based on genomic sequence of Shinorhizobium meliloti Sequence analysis of the cloned DNA fragment revealed one open-reading frame of 1059 bp and 386 amino acids. This putative D-TA gene was cloned into NdeI and EcoRI (pEnsi His-tag sequence or BamHI (pEnsi-DTA[2]) sequence of the pET21(a) vector. The expression level of the cloned gene was extremely overexpressed by E. coli BL21(DE3) transformed with pEnsi-DTA[1] compared to E. coli BL21(DE3) transformed with pEnsi-DTA[2]. When the cells expressing the wild used for D-TA enzyme activity, 12 mM glycine was successfully detected in HPLC analysis. Moreover, the whole cells harbouring the recombinant D-TA was able to synthesize D-erythro of 0.6 mg/ml in a batch reaction.

Keywords: About four key words or phrases in alphabetical order, separated by commas.

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84 Identification Common Microbes Observed on Polyester Tufting

Authors: A. Ashjaran, M.E. Yazdanshenas, R. Ghazi Saeidi, S. Moghadamifar

Abstract:

Tufting carpet is a very suitable substrate for growing microorganism such as pathogenic microbes, due to the direct touch with human body, long washing periods and laying on the floor; in fact there are 3 major problems: To risk human health, Prepare bad odors and Destruction of the products.. In the presented research, for investigation of presence most common microbes on polyester tufting, first goods laid in a public place (in the corridor fair) for 30 days and the existence of some microbes were investigate on it with two methods of enrichment in nutrient environments such as thioglycolate and noutrunt brath, and shake the dust off the polyester tufting onto cultivation mediums such as blood agar and noutrunt agar. After the microorganism colonics are grown, the colonies were separated and six microbial tests such as cataloes and sitrat were carried out in five phases on the colonics for identifying the varieties of bacteria. As a result of tests, 5 type of bacteria, such as Escherichia coli, staphylococcus saprophytic as were identified. Each of the mentioned bacteria can be seriously harmful for the heath of human.

Keywords: Microorganisms, Polyester tufting, Escherichia coli, Staphylococcus saprophytic, Blood agar, Thioglycolate

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83 PCR based Detection of Food Borne Pathogens

Authors: Archana Panchapakesan Iyer, Taha Abdullah Kumosani

Abstract:

Many high-risk pathogens that cause disease in humans are transmitted through various food items. Food-borne disease constitutes a major public health problem. Assessment of the quality and safety of foods is important in human health. Rapid and easy detection of pathogenic organisms will facilitate precautionary measures to maintain healthy food. The Polymerase Chain Reaction (PCR) is a handy tool for rapid detection of low numbers of bacteria. We have designed gene specific primers for most common food borne pathogens such as Staphylococci, Salmonella and E.coli. Bacteria were isolated from food samples of various food outlets and identified using gene specific PCRs. We identified Staphylococci, Salmonella and E.coli O157 using gene specific primers by rapid and direct PCR technique in various food samples. This study helps us in getting a complete picture of the various pathogens that threaten to cause and spread food borne diseases and it would also enable establishment of a routine procedure and methodology for rapid identification of food borne bacteria using the rapid technique of direct PCR. This study will also enable us to judge the efficiency of present food safety steps taken by food manufacturers and exporters.

Keywords: food borne pathogens, PCR, food safety, rapiddetection.

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82 Effects and Mechanization of a High Gradient Magnetic Separation Process for Particulate and Microbe Removal from Ballast Water

Authors: Zhijun Ren, Zhang Lin, Zhao Ye, Zuo Xiangyu, Mei Dongxing

Abstract:

As a pretreatment process of ballast water treatment, the performance of high gradient magnetic separation (HGMS) technology for the removal of particulates and microorganisms was studied. The results showed that HGMS process could effectively remove suspended particles larger than 5 µm and had ability to resist impact load. Microorganism could also be effectively removed by HGMS process, and the removal effect increased with increasing magnetic field strength. The maximum removal rates for Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were 4016.1% and 9675.3% higher, respectively, than without the magnetic field. In addition, the superoxide dismutase (SOD) activity of the microbes decreased by 32.2% when the magnetic field strength was 15.4 mT for 72 min. The microstructure of the stainless steel wool was investigated, and the results showed that particle removal by HGMS has common function by the magnetic force of the high-strength, high-gradient magnetic field on weakly magnetic particles in the water, and on the stainless steel wool.

Keywords: HGMS, particulates, superoxide dismutase activity, steel wool magnetic medium.

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