Search results for: Bacillus subtilis ATCC 21332
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 88

Search results for: Bacillus subtilis ATCC 21332

28 LIFirr with an Indicator of Microbial Activity in Paraffinic Oil

Authors: M. P. Casiraghi, C. M. Quintella, P. Almeida

Abstract:

Paraffinic oils were submitted to microbial action. The microorganisms consisted of bacteria of the genera Pseudomonas sp. and Bacillus lincheniforms. The alterations in interfacial tension were determined using a tensometer and applying the hanging drop technique at room temperature (299 K ±275 K). The alteration in the constitution of the paraffins was evaluated by means of gas chromatography. The microbial activity was observed to reduce interfacial tension by 54 to 78%, as well as consuming the paraffins C19 to C29 and producing paraffins C36 to C44. The LIFirr technique made it possible to determine the microbial action quickly.

Keywords: Paraffins, biosurfactants, LIFirr.

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27 Kinetics Study for the Recombinant Cellulosome to the Degradation of Chlorella Cell Residuals

Authors: C.-C. Lin, S.-C. Kan, C.-W. Yeh, C.-I Chen, C.-J. Shieh, Y.-C. Liu

Abstract:

In this study, lipid-deprived residuals of microalgae were hydrolyzed for the production of reducing sugars by using the recombinant Bacillus cellulosome, carrying eight genes from the Clostridium thermocellum ATCC27405. The obtained cellulosome was found to exist mostly in the broth supernatant with a cellulosome activity of 2.4 U/mL. Furthermore, the Michaelis-Menten constant (Km) and Vmax of cellulosome were found to be 14.832 g/L and 3.522 U/mL. The activation energy of the cellulosome to hydrolyze microalgae LDRs was calculated as 32.804 kJ/mol.

Keywords: Lipid-deprived residuals of microalgae, cellulosome, cellulose, reducing sugars, kinetics.

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26 In vitro Anti-tubercular Screening of Newly Synthesized Benzimidazole Derivatives

Authors: M. Shahar Yar, M. Mustaqeem Abdullah, Jaseela Majeed

Abstract:

A series of 1-(1H-benzimidazol-2-yl)-3-(substituted phenyl)-2-propen-1-one were allowed to react with hydrazine hydrate and phenyl hydrazine in submitted reactions to get pyrazoline and phenyl pyrazoline derivatives. All the compounds entered for screening at the Tuberculosis Antimicrobial Acquisition and Coordinating Facility (TAACF) for their in vitro antibacterial activity against Mycobacterium tuberculosis H37Rv strain (ATCC 27294) using Microplate Alamar Blue Assay (MABA) susceptibility test. The results expressed as MIC (minimum inhibitory concentration) in μg/mL. Among the fifteen compounds, eight compounds were found to have MIC values less than 10 μg/mL. These were subjected for cytotoxicity assay in VERO cells to determine CC50 (cytotoxic concentration 50%) values and finally SI (Selectivity Index) were calculated. Compound (XV) 2-[5-(4- fluorophenyl)-1-phenyl-4,5-dihydro-1H-3-pyrazolyl]-1Hbenzimidazole was considered the best candidate of the series that could be a good starting point to develop new lead compounds in the fight against tuberculosis.

Keywords: anti-tubercular activity, benzimidazole, pyrazoline.

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25 Effect of Organic Matter and Biofertilizers on Chickpea Quality and Biological Nitrogen Fixation

Authors: Khosro Mohammadi, Amir Ghalavand, Majid Aghaalikhani

Abstract:

In order to evaluation the effects of soil organic matter and biofertilizer on chickpea quality and biological nitrogen fixation, field experiments were carried out in 2007 and 2008 growing seasons. In this research the effects of different strategies for soil fertilization were investigated on grain yield and yield component, minerals, organic compounds and cooking time of chickpea. Experimental units were arranged in split-split plots based on randomized complete blocks with three replications. Main plots consisted of (G1): establishing a mixed vegetation of Vicia panunica and Hordeum vulgare and (G2): control, as green manure levels. Also, five strategies for obtaining the base fertilizer requirement including (N1): 20 t.ha-1 farmyard manure; (N2): 10 t.ha-1 compost; (N3): 75 kg.ha-1 triple super phosphate; (N4): 10 t.ha-1 farmyard manure + 5 t.ha-1 compost and (N5): 10 t.ha-1 farmyard manure + 5 t.ha-1 compost + 50 kg.ha-1 triple super phosphate were considered in sub plots. Furthermoree four levels of biofertilizers consisted of (B1): Bacillus lentus + Pseudomonas putida; (B2): Trichoderma harzianum; (B3): Bacillus lentus + Pseudomonas putida + Trichoderma harzianum; and (B4): control (without biofertilizers) were arranged in sub-sub plots. Results showed that integrating biofertilizers (B3) and green manure (G1) produced the highest grain yield. The highest amounts of yield were obtained in G1×N5 interaction. Comparison of all 2-way and 3-way interactions showed that G1N5B3 was determined as the superior treatment. Significant increasing of N, P2O5, K2O, Fe and Mg content in leaves and grains emphasized on superiority of mentioned treatment because each one of these nutrients has an approved role in chlorophyll synthesis and photosynthesis abilities of the crops. The combined application of compost, farmyard manure and chemical phosphorus (N5) in addition to having the highest yield, had the best grain quality due to high protein, starch and total sugar contents, low crude fiber and reduced cooking time.

Keywords: chickpea, biofertilizer, nitrogen fixation.

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24 Activation of Prophenoloxidase during Bacterial Injection into the Desert Locust, Schistocerca Gregaria

Authors: Shaiemaa, A. Momen, Dalia, A.M. Salem, Emad, M.S. Barakat, Mohamed, S. Salama

Abstract:

The present study has been conducted to characterize the prophenoloxidase (PPO) system of the desert locust, Schistocerca gregaria following injection of Bacillus thuringiensis kurstaki (Bt). The bulk of PPO system was associated with haemocytes and a little amount was found in plasma. This system was activated by different activators such as laminarin, lipopolysaccharide (LPS) and trypsin suggesting that the stimulatory mechanism may involve an enzyme cascade of one or more associated molecules. These activators did not activate all the molecules of the cascade. Presence of phenoloxidase activity (PO) coincides with the appearance of protein band with molecular weight (MW) 70.154 KD (Kilo Dalton).

Keywords: Schistocerca gregaria, haemolymph, proteins, prophenoloxidase system, phenoloxidase

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23 Pt(IV) Complexes with Polystrene-bound Schiff Bases as Antimicrobial Agent: Synthesis and Characterization

Authors: Dilek Nartop, Nurşen Sarı, Hatice Öğütçü

Abstract:

Novel polystrene-bound Schiff bases and their Pt(IV) complexes have been prepared from condensation reaction of polystyrene-A-NH2 with 2-hydroxybenzaldehyde and 5-fluoro-3- bromo-2-hydroxybenzaldehyde. The structures of Pt(IV) complexes with polystyrene including Schiff bases have been determined by elemental analyses, magnetic susceptibility, IR, 1H-NMR, UV-vis, TG/DTA and AAS. The antibacterial and antifungal activities of the synthesized compounds have been studied by the well-diffusion method against some selected microorganisms: (Bacillus cereus spp., Listeria monocytogenes 4b, Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermis, Brucella abortus, Escherichia coli, Pseudomonas putida spp., Shigella dysenteria type 10, Salmonella typhi H).

Keywords: Polymer-bound Schiff bases, polystyrene-A-NH2, Pt(IV) complexes, biological activity.

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22 Tolerance of Heavy Metals by Gram Positive Soil Bacteria

Authors: I. V. N. Rathnayake, Mallavarapu Megharaj, Nanthi Bolan, Ravi Naidu

Abstract:

With the intention of screening for heavy metal tolerance, a number of bacteria were isolated and characterized from a pristine soil. Two Gram positive isolates were identified as Paenibacillus sp. and Bacillus thuringeinsis. Tolerance of Cd2+, Cu2+ and Zn2+ by these bacteria was studied and found that both bacteria were highly sensitive to Cu2+ compared to other two metals. Both bacteria showed the same pattern of metal tolerance in the order Zn+ > Cd2+ > Cu2+. When the metal tolerance in both bacteria was compared, Paenibacillus sp. showed the highest sensitivity to Cu2+ where as B. thuringiensis showed highest sensitivity to Cd2+ and Zn2+ .These findings revealed the potential of Paenibacillus sp. in developing a biosensor to detect Cu2+ in environmental samples.

Keywords: Heavy metals, bacteria, soil, tolerance.

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21 Partial Purification of Cytotoxic Peptides against Gastric Cancer Cells from Protein Hydrolysate of Euphorbia hirta Linn.

Authors: S. Yodyingyong, C. Chaichana, C. Nuchsuk, S. Roytrakul, N. P. T-Thienprasert, S. Ratanapo

Abstract:

Protein hydrolysates prepared from a number of medicinal plants are promising sources of various bioactive peptides. In this work, proteins from dried whole plant of Euphorbia hirta Linn. were extracted and digested with pepsin for 12h. The hydrolysates of lesser than 3 KDa were fractionated by a cut-off membrane. The peptide hydrolysate was then purified by an anion-exchange chromatography on DEAE-Sephacel™ column and reverse-phase chromatography on Sep-pak C18 column, respectively. The cytotoxic effect of each peptide fraction against a gastric carcinoma cell line (KATO-III, ATCC No. HTB103) was investigated using colorimetric MTT viability assay. A human liver cell line (Chang Liver, CLS No. 300139) was used as a control normal cell line. Two purified peptide peaks, peak l and peak ll at 100µg peptides mL-1 affected cell viability of the gastric cancer cell lines to 63.85±4.94 and 66.92±6.46%, respectively. Our result showed for the first time that the peptide fractions derived from protein hydrolysate of Euphorbia hirta Linn. have anti-gastric cancer activity, which offers a potential novel and natural anti-gastric cancer remedy.

Keywords: Cytotoxic, peptides, Euphorbia hirta Linn., gastric carcinoma.

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

Authors: Salem El-t. Ashoor, Ibtisam M. Ediab

Abstract:

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.

Keywords: (1:10-phenanthroline) (L1), 4-4-amino phenazone (L2), diphenyl carbizon (L3), DFT study, antibacterial.

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19 Probiotics’ Antibacterial Activity on Beef and Camel Minced Meat at Altered Ranges of Temperature

Authors: Rania Samir Zaki

Abstract:

Because of their inhibitory effects, selected probiotic Lactobacilli may be used as antimicrobial against some hazardous microorganisms responsible for spoilage of fresh minced beef (cattle) minced meat and camel minced meat. Lactic acid bacteria were isolated from camel meat. These included 10 isolates; 1 Lactobacillus fermenti, 4 Lactobacillus plantarum, 4 Lactobacillus pulgaricus, 3 Lactobacillus acidophilus and 1 Lactobacillus brevis. The most efficient inhibitory organism was Lactobacillus plantarum which can be used as a propiotic with antibacterial activity. All microbiological analyses were made at the time 0, first day and the second day at altered ranges of temperature [4±2 ⁰C (chilling temperature), 25±2 ⁰C, and 38±2 ⁰C]. Results showed a significant decrease of pH 6.2 to 5.1 within variant types of meat, in addition to reduction of Total Bacterial Count, Enterococci, Bacillus cereus and Escherichia coli together with the stability of Coliforms and absence of Staphylococcus aureus.

Keywords: Antibacterial, camel meat, inhibition, probiotics.

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18 Evaluation of the Microbiological, Chemical and Sensory Quality of Carp Processed by the Sous Vide Method

Authors: Özlem Pelin Can

Abstract:

This study evaluated the microbiological quality and the sensory characteristics of carp fillets processed by the sousvide method when stored at 2 and 10 °C. Four different combinations of sauced–storage were studied then stored at 2 or 10 °C was evaluate periodically sensory, microbiological and chemical quality. Batches stored at 2 °C had lower growth rates of mesophiles and psychrotrophs. Moreover, these counts decreased by increasing the heating temperature and time. Staphylococcus aureus, Bacillus cereus, Clostridium perfringens and Listeria monocytogenes were not found in any of the samples. The heat treatment of 90 °C for 15 min and sauced was the most effective to ensure the safety and extend the shelf-life of sousvide carp preserving its sensory characteristics. This study establishes the microbiological quality of sous vide carp and emphasizes the relevance of the raw materials, heat treatment and storage temperature to ensure the safety of the product.

Keywords: Sous- vide methods, carp, sauce, microbiological, chemical and sensory quality

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17 Solid State Fermentation of Cassava Peel with Trichoderma viride (ATCC 36316) for Protein Enrichment

Authors: Olufunke O. Ezekiel, Ogugua C. Aworh

Abstract:

Solid state fermentation of cassava peel with emphasis on protein enrichment using Trichoderma viride was evaluated. The effect of five variables: moisture content, pH, particle size (p), nitrogen source and incubation temperature; on the true protein and total sugars of cassava peel was investigated. The optimum fermentation period was established to be 8 days. Total sugars were 5-fold higher at pH 6 relative to pH 4 and 7-fold higher when cassava peels were fermented at 30oC relative to 25oC as well as using ammonium sulfate as the nitrogen source relative to urea or a combination of both. Total sugars ranged between 123.21mg/g at 50% initial moisture content to 374mg/g at 60% and from 190.59mg/g with particle size range of 2.00>p>1.41mm to 310.10mg/g with 4.00>p>3.35mm.True protein ranged from 229.70 mg/g at pH 4 to 284.05 mg/g at pH 6; from 200.87 mg/g with urea as nitrogen source and to 254.50mg/g with ammonium sulfate; from 213.82mg/g at 50% initial moisture content to 254.50mg/g at 60% moisture content, from 205.75mg/g in cassava peel with 5.6>p> 4.75mm to 268.30 in cassava peel with particle size 4.00>p>3.35mm, from 207.57mg/g at 25oC to 254.50mg/g at 30oC Cassava peel with particle size 4.00>p>3.35 mm and initial moisture content of 60% at pH 6.0, 30oC incubation temperature with ammonium sulfate (10g N / kg substrate) was most suitable for protein enrichment with Trichoderma viride. Crude protein increased from 4.21 % in unfermented cassava peel samples to 10.43 % in fermented samples.

Keywords: Cassava peel, Solid state fermentation, Trichoderma viride, Total sugars, True protein.

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16 Synthesis of Peptide Amides using Sol-Gel Immobilized Alcalase in Batch and Continuous Reaction System

Authors: L. N. Corîci, A. E. Frissen, D -J. Van Zoelen, I. F. Eggen, F. Peter, C. M. Davidescu, C. G. Boeriu

Abstract:

Two commercial proteases from Bacillus licheniformis (Alcalase 2.4 L FG and Alcalase 2.5 L, Type DX) were screened for the production of Z-Ala-Phe-NH2 in batch reaction. Alcalase 2.4 L FG was the most efficient enzyme for the C-terminal amidation of Z-Ala-Phe-OMe using ammonium carbamate as ammonium source. Immobilization of protease has been achieved by the sol-gel method, using dimethyldimethoxysilane (DMDMOS) and tetramethoxysilane (TMOS) as precursors (unpublished results). In batch production, about 95% of Z-Ala-Phe-NH2 was obtained at 30°C after 24 hours of incubation. Reproducibility of different batches of commercial Alcalase 2.4 L FG preparations was also investigated by evaluating the amidation activity and the entrapment yields in the case of immobilization. A packed-bed reactor (0.68 cm ID, 15.0 cm long) was operated successfully for the continuous synthesis of peptide amides. The immobilized enzyme retained the initial activity over 10 cycles of repeated use in continuous reactor at ambient temperature. At 0.75 mL/min flow rate of the substrate mixture, the total conversion of Z-Ala-Phe-OMe was achieved after 5 hours of substrate recycling. The product contained about 90% peptide amide and 10% hydrolysis byproduct.

Keywords: packed-bed reactor, peptide amide, protease, sol-gel immobilization.

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15 Control of Staphylococcus aureus in Meat System by in situ and ex situ Bacteriocins from Lactobacillus sakei and Pediococcus spp.

Authors: M. Naimi, M. B. Khaled

Abstract:

The present study consisted of an applied test in meat system to assess the effectiveness of three bio agents bacteriocinproducing strains: Lm24: Lactobacillus sakei, Lm14and Lm25: Pediococcus spp. Two tests were carried out: The ex situ test was intended for three batches added with crude bacteriocin solutions at 12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the in situ one consisted of four batches; three of them inoculated with one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The two used tests were done in the presence of the pathogen St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch served as a positive or a negative control was used too. The incubation was performed at 7°C. Total viable counts, staphylococci and lactic acid bacteria, at the beginning and at selected times with interval of three days were enumerated. Physico-chemical determinations (except for in situ test): pH, dry mater, sugars, fat and total protein, at the beginning and at end of the experiment, were done, according to the international norms. Our results confirmed the ex situ effectiveness. Furthermore, the batches affected negatively the total microbial load over the incubation days, and showed a significant regression in staphylococcal load at day seven, for Lm14, Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed that, at the last day of culture, staphylococcal load was nil for the three batches. In the in situ test, the cultures displayed less inhibitory attitude and recorded a decrease in staphylococcal load, for Lm14, Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore, physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%, 0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%, 5.37%, 5.5%. This study suggests that the use of selected strains as Lm25 could lead to the best results and would help in preserving and extending the shelf life of lamb meat.

Keywords: Biocontrol, in situ and ex situ, meat system, St. aureus, Lactobacillus sakei, Pediococcus spp.

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14 Cadmium Filter Cake of a Hydrometallurgical Zinc Smelter as a New Source for the Biological Synthesis of CdS Quantum Dots

Authors: Mehran Bakhshi, Mohammad Raouf Hosseini, Mohammadhosein Rahimi

Abstract:

The cadmium sulfide nanoparticles were synthesized from the nickel-cadmium cake of a hydrometallurgical zinc producing plant and sodium sulfide as Cd2+ and S-2 sources, respectively. Also, the synthesis process was performed by using the secretions of Bacillus licheniformis as bio-surfactant. Initially, in order to obtain a cadmium rich solution, two following steps were carried out: 1) Alkaline leaching for the removal of zinc oxide from the cake, and 2) acidic leaching to dissolve cadmium from the remained solid residue. Afterward, the obtained CdSO4 solution was used for the nanoparticle biosynthesis. Nanoparticles were characterized by the energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD) to confirm the formation of CdS crystals with cubic structure. Also, transmission electron microscopy (TEM) was applied to determine the particle sizes which were in 2-10 nm range. Moreover, the presence of the protein containing bio-surfactants was approved by using infrared analysis (FTIR). In addition, the absorbance below 400 nm confirms quantum particles’ size. Finally, it was shown that valuable CdS quantum dots could be obtained from the industrial waste products via environment-friendly biological approaches.

Keywords: Biosynthesis, cadmium cake, cadmium sulfide, nanoparticle, zinc smelter.

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13 Antimicrobial Activity and Phytochemicals Screening of Jojoba (Simmondsia chinensis) Root Extracts and Latex

Authors: Ferial M. Abu-Salem, Hayam M. Ibrahim

Abstract:

Plants are rich sources of bioactive compounds. In this study the photochemical screening of hexane, ethanolic and aqueous extracts of roots and latex of jojoba (Simmondsia chinensis) plant revealed the presence of saponins, tannins, alkaloids, steroids and glycosides. Ethanolic extract was found to be richer in these metabolites than hexane, aqueous extracts and latex. The extracts and latex displayed effective antimicrobial activity against Salmonella typhimurium, Bacillus cereus, Clostridium perfringens, Staphylococcus aureus, Escherichia coli, Candida albicans and Aspergillus flavus. The increase in volume of the extracts and latex caused more activity, as shown by zones of inhibition. Candida albicans growth was inhibited only by hexane extract. Jojoba latex was not effective against Candida albicans at 0.1 and 0.5 ml extracts concentration but showed 5mm zone of inhibition at (1.0 ml). Lower volume (0.1ml) of latex encouraged Aspergillus flavus growth, while at (1.00 ml) reduced its mycelial growth. Thus, jojoba root extracts and latex can be of potential natural antimicrobial agents.

Keywords: Antimicrobial activity, Jojoba (Simmondsia chinensis), latex, photochemical, root Extracts.

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12 Characterization of the Microbial Induced Carbonate Precipitation Technique as a Biological Cementing Agent for Sand Deposits

Authors: Sameh Abu El-Soud, Zahra Zayed, Safwan Khedr, Adel M. Belal

Abstract:

The population increase in Egypt is urging for horizontal land development which became a demand to allow the benefit of different natural resources and expand from the narrow Nile valley. However, this development is facing challenges preventing land development and agriculture development. Desertification and moving sand dunes in the west sector of Egypt are considered the major obstacle that is blocking the ideal land use and development. In the proposed research, the sandy soil is treated biologically using Bacillus pasteurii bacteria as these bacteria have the ability to bond the sand partials to change its state of loose sand to cemented sand, which reduces the moving ability of the sand dunes. The procedure of implementing the Microbial Induced Carbonate Precipitation Technique (MICP) technique is examined, and the different factors affecting on this process such as the medium of bacteria sample preparation, the optical density (OD600), the reactant concentration, injection rates and intervals are highlighted. Based on the findings of the MICP treatment for sandy soil, conclusions and future recommendations are reached.

Keywords: Soil stabilization, biological treatment, MICP, sand cementation.

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11 Chemical Composition of Essential Oil and in vitro Antibacterial and Anticancer Activity of the Hydroalcolic Extract from Coronilla varia

Authors: Dehpour A. A., Eslami B., Rezaie S., Hashemian S. F., Shafie F., Kiaie M.

Abstract:

The aims of study were investigation on chemical composition essential oil and the effect of extract of Coronilla varia on antimicrobial and cytotoxicity activity. The essential oils of Coronilla varia is obtained by hydrodistillation and analyzed by (GC/MS) for determining their chemical composition and identification of their components. Antibacterial activity of plant extract was determined by disc diffusion method and anticancer activity measured by MTT assay. The major components in essential oil were Caryophyllene Oxide (60.19%), Alphacadinol (4.13%) and Homoadantaneca Robexylic Acid (3.31%). The extracts from Coronilla varia had interesting activity against Proteus mirabilis in the concentration of 700 μg/disc and did not show any activity against Staphylococus aureus, Bacillus subtillis, Klebsiella pneumonia and Entrobacter cloacae. The positive control, Ampicillin, Chloramphenicol and Cenphalothin had shown zone of inhibition resistant all bacteria. The ethanol extract of Corohilla varia inhibited on MCF7 cell lines. IC50 0.6(mg/ml) was the optimum concentration of extract from Coronilla varia inhibition of cell line growth. The MCF7 cancer cell line and Proteus mirabilis were more sensitive to Coronilla varia ethanol extract.

Keywords: Coronilla varia, Essential oil, Antibacterial, Anticancer, HeLa cell line.

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10 An Advanced Approach Based on Artificial Neural Networks to Identify Environmental Bacteria

Authors: Mauro Giacomini, Stefania Bertone, Federico Caneva Soumetz, Carmelina Ruggiero

Abstract:

Environmental micro-organisms include a large number of taxa and some species that are generally considered nonpathogenic, but can represent a risk in certain conditions, especially for elderly people and immunocompromised individuals. Chemotaxonomic identification techniques are powerful tools for environmental micro-organisms, and cellular fatty acid methyl esters (FAME) content is a powerful fingerprinting identification technique. A system based on an unsupervised artificial neural network (ANN) was set up using the fatty acid profiles of standard bacterial strains, obtained by gas-chromatography, used as learning data. We analysed 45 certified strains belonging to Acinetobacter, Aeromonas, Alcaligenes, Aquaspirillum, Arthrobacter, Bacillus, Brevundimonas, Enterobacter, Flavobacterium, Micrococcus, Pseudomonas, Serratia, Shewanella and Vibrio genera. A set of 79 bacteria isolated from a drinking water line (AMGA, the major water supply system in Genoa) were used as an example for identification compared to standard MIDI method. The resulting ANN output map was found to be a very powerful tool to identify these fresh isolates.

Keywords: Cellular fatty acid methyl esters, environmental bacteria, gas-chromatography, unsupervised ANN.

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9 Biological Effects of a Carbohydrate-Binding Protein from an Annelid, Perinereis nuntia Against Human and Phytopathogenic Microorganisms

Authors: Sarkar M. A. Kawsar, Sarkar M. A. Mamun, Md S. Rahman, Hidetaro Yasumitsu, Yasuhiro Ozeki

Abstract:

Lectins have a good scope in current clinical microbiology research. In the present study evaluated the antimicrobial activities of a D-galactose binding lectin (PnL) was purified from the annelid, Perinereis nuntia (polychaeta) by affinity chromatography. The molecular mass of the lectin was determined to be 32 kDa as a single polypeptide by SDS-PAGE under both reducing and non-reducing conditions. The hemagglutinating activity of the PnL showed against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-Gal, GalNAc, Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro antibacterial screening studies against 11 gram-positive and gram-negative microorganisms. From the screening results, it was revealed that PnL exhibited significant antibacterial activity against gram-positive bacteria. Bacillus megaterium showed the highest growth inhibition by the lectin (250 μg/disc). However, PnL did not inhibit the growth of gram-negative bacteria such as Vibrio cholerae and Pseudomonas sp. PnL was also examined for in vitro antifungal activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited the mycelial growth of Alternaria alternata (24.4%). These results indicate that future findings of lectin applications obtained from annelids may be of importance to life sciences.

Keywords: Perinereis nuntia, Lectin, Inhibition zone, Mycelial growth.

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8 Microbiological and Physicochemical Studies of Wetland Soils in Eket, Nigeria

Authors: Ime R. Udotong, Ofonime U. M. John, Justina I. R. Udotong

Abstract:

The microbiological and physicochemical characteristics of wetland soils in Eket Local Government Area were studied between May 2001 and June 2003. Total heterotrophic bacterial counts (THBC), total fungal counts (TFC), and total actinomycetes counts (TAC) were determined from soil samples taken from four locations at two depths in the wet and dry seasons. Microbial isolates were characterized and identified. Particle size and chemical parameters were also determined using standard methods. THBC ranged from 5.2 (+0.17) x106 to 1.7 (+0.18) x107 cfu/g and from 2.4 (+0.02) x106 to 1.4 (+0.04) x107cfu/g in the wet and dry seasons, respectively. TFC ranged from 1.8 (+0.03) x106 to 6.6 (+ 0.18) x106 cfu/g and from 1.0 (+0.04) x106 to 4.2 (+ 0.01) x106 cfu/g in the wet and dry seasons, respectively .TAC ranged from 1.2 (+0.53) x106 to 6.0 (+0.05) x106 cfu/g and from 0.6 (+0.01) x106 to 3.2 (+ 0.12) x106 cfu/g in the wet and dry season, respectively. Acinetobacter, Alcaligenes, Arthrobacter, Bacillus, Beijerinckja, Enterobacter, Micrococcus, Flavobacterium, Serratia, Enterococcus, and Pseudomonas species were predominant bacteria while Aspergillus, Fusarium, Mucor, Penicillium, and Rhizopus were the dominant fungal genera isolated. Streptomyces and Norcadia were the actinomycetes genera isolated. The particle size analysis showed high sand fraction but low silt and clay. The pH and % organic matter were generally acidic and low, respectively at all locations. Calcium dominated the exchangeable bases with low electrical conductivity and micronutrients. These results provide the baseline data of Eket wetland soils for its management for sustainable agriculture.

Keywords: Wetland soils, Microbial counts, physicochemicalcharacteristics, Sustainable agriculture.

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7 Wheat Bran Carbohydrates as Substrate for Bifidobacterium lactis Development

Authors: V. Radenkovs, D. Klava, K. Juhnevica

Abstract:

The present study addresses problems and solutions related to new functional food production. Wheat (Triticum aestivum L) bran obtained from industrial mill company “Dobeles dzirnavieks”, was used to investigate them as raw material like nutrients for Bifidobacterium lactis Bb-12. Enzymatic hydrolysis of wheat bran starch was carried out by α-amylase from Bacillus amyloliquefaciens (Sigma Aldrich). The Viscozyme L purchased from (Sigma Aldrich) were used for reducing released sugar. Bifidibacterium lactis Bb-12 purchased from (Probio-Tec® CHR Hansen) was cultivated in enzymatically hydrolysed wheat bran mash. All procedures ensured the number of active Bifidobacterium lactis Bb-12 in the final product reached 105 CFUg-1. After enzymatic and bacterial fermentations sample were freeze dried for analysis of chemical compounds. All experiments were performed at Faculty of Food Technology of Latvia University of Agriculture in January- March 2013. The obtained results show that both types of wheat bran (enzymatically treated and non-treated) influenced the fermentative activity and number of Bifidibacterium lactis Bb-12 viable in wheat bran mash. Amount of acidity strongly increase during the wheat bran mash fermentation. The main objective of this work was to create low-energy functional enzymatically and bacterially treated food from wheat bran using enzymatic hydrolysis of carbohydrates and following cultivation of Bifidobacterium lactis Bb-12.

Keywords: Viscozyme L, α-amylase, Bifidobacterium lactis, fermented wheat bran.

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6 Isolation of a Bacterial Community with High Removal Efficiencies of the Insecticide Bendiocarb

Authors: Eusebio A. Jiménez-Arévalo, Deifilia Ahuatzi-Chacón, Juvencio Galíndez-Mayer, Cleotilde Juárez-Ramírez, Nora Ruiz-Ordaz

Abstract:

Bendiocarb is a known toxic xenobiotic that presents acute and chronic risks for freshwater invertebrates and estuarine and marine biota; thus, the treatment of water contaminated with the insecticide is of concern. In this paper, a bacterial community with the capacity to grow in bendiocarb as its sole carbon and nitrogen source was isolated by enrichment techniques in batch culture, from samples of a composting plant located in the northeast of Mexico City. Eight cultivable bacteria were isolated from the microbial community, by PCR amplification of 16 rDNA; Pseudoxanthomonas spadix (NC_016147.2, 98%), Ochrobacterium anthropi (NC_009668.1, 97%), Staphylococcus capitis (NZ_CP007601.1, 99%), Bosea thiooxidans. (NZ_LMAR01000067.1, 99%), Pseudomonas denitrificans. (NC_020829.1, 99%), Agromyces sp. (NZ_LMKQ01000001.1, 98%), Bacillus thuringiensis. (NC_022873.1, 97%), Pseudomonas alkylphenolia (NZ_CP009048.1, 98%). NCBI accession numbers and percentage of similarity are indicated in parentheses. These bacteria were regarded as the isolated species for having the best similarity matches. The ability to degrade bendiocarb by the immobilized bacterial community in a packed bed biofilm reactor, using as support volcanic stone fragments (tezontle), was evaluated. The reactor system was operated in batch using mineral salts medium and 30 mg/L of bendiocarb as carbon and nitrogen source. With this system, an overall removal efficiency (ηbend) rounding 90%, was reached.

Keywords: Bendiocarb, biodegradation, biofilm reactor, carbamate insecticide.

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5 Use of Corn Stover for the Production of 2G Bioethanol, Enzymes and Xylitol under a Biorefinery Concept

Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

Abstract:

The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever growing accepted option in the way to the development of biorefinery complexes. In the Mexican state of Sinaloa, a significant amount of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second generation bioethanol (2G), enzymes and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our work group. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU/mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: Biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol.

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4 Density of Hydrocarbonoclastic Bacteria and Polycyclic Aromatic Hydrocarbon Accumulation in Iko River Mangrove Ecosystem, Nigeria

Authors: Ime R. Udotong, Samuel I. Eduok, Joseph P. Essien, Basil N. Ita

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Sediment and mangrove root samples from Iko River Estuary, Nigeria were analyzed for microbial and polycyclic aromatic hydrocarbon (PAH) content. The total heterotrophic bacterial (THB) count ranged from 1.1x107 to 5.1 x107 cfu/g, total fungal (TF) count ranged from 1.0x106 to 2.7x106 cfu/g, total coliform (TC) count ranged from 2.0x104 to 8.0x104cfu/g while hydrocarbon utilizing bacterial (HUB) count ranged from 1.0x 105 to 5.0 x 105cfu/g. There was a range of positive correlation (r = 0.72 to 0.93) between THB count and total HUB count, respectively. The organisms were Staphylococcus aureus, Bacillus cereus, Flavobacterium breve, Pseudomonas aeruginosa, Erwinia amylovora, Escherichia coli, Enterobacter sp, Desulfovibrio sp, Acinetobacter iwoffii, Chromobacterium violaceum, Micrococcus sedentarius, Corynebacterium sp, and Pseudomonas putrefaciens. The PAH were Naphthalene, 2-Methylnaphthalene, Acenapthylene, Acenaphthene, Fluorene, Phenanthene, Anthracene, Fluoranthene, Pyrene, Benzo(a)anthracene, Chrysene, Benzo(b)fluoranthene, Benzo(k)fluoranthene, Benzo(a)pyrene, Dibenzo(a,h)anthracene, Benzo(g,h,l)perylene ,Indeno(1,2,3-d)pyrene with individual PAH concentrations that ranged from 0.20mg/kg to 1.02mg/kg, 0.20mg/kg to 1.07mg/kg and 0.2mg/kg to 4.43mg/kg in the benthic sediment, epipellic sediment and mangrove roots, respectively. Total PAH ranged from 6.30 to 9.93mg/kg, 6.30 to 9.13mg/kg and 9.66 to 16.68mg/kg in the benthic sediment, epipellic sediment and mangrove roots, respectively. The high concentrations in the mangrove roots are indicative of bioaccumulation of the pollutant in the plant tissue. The microorganisms are of ecological significance and the detectable quantities of polycyclic aromatic hydrocarbon could be partitioned and accumulated in tissues of infaunal and epifaunal organisms in the study area.

Keywords: Hydrocarbonoclastic bacteria, Iko River estuary, Mangrove, Polycyclic aromatic hydrocarbon.

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3 Identification of Promiscuous Epitopes for Cellular Immune Responses in the Major Antigenic Protein Rv3873 Encoded by Region of Difference 1 of Mycobacterium tuberculosis

Authors: Abu Salim Mustafa

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Rv3873 is a relatively large size protein (371 amino acids in length) and its gene is located in the immunodominant genomic region of difference (RD)1 that is present in the genome of Mycobacterium tuberculosis but deleted from the genomes of all the vaccine strains of Bacillus Calmette Guerin (BCG) and most other mycobacteria. However, when tested for cellular immune responses using peripheral blood mononuclear cells from tuberculosis patients and BCG-vaccinated healthy subjects, this protein was found to be a major stimulator of cell mediated immune responses in both groups of subjects. In order to further identify the sequence of immunodominant epitopes and explore their Human Leukocyte Antigen (HLA)-restriction for epitope recognition, 24 peptides (25-mers overlapping with the neighboring peptides by 10 residues) covering the sequence of Rv3873 were synthesized chemically using fluorenylmethyloxycarbonyl chemistry and tested in cell mediated immune responses. The results of these experiments helped in the identification of an immunodominant peptide P9 that was recognized by people expressing varying HLA-DR types. Furthermore, it was also predicted to be a promiscuous binder with multiple epitopes for binding to HLA-DR, HLA-DP and HLA-DQ alleles of HLA-class II molecules that present antigens to T helper cells, and to HLA-class I molecules that present antigens to T cytotoxic cells. In addition, the evaluation of peptide P9 using an immunogenicity predictor server yielded a high score (0.94), which indicated a greater probability of this peptide to elicit a protective cellular immune response. In conclusion, P9, a peptide with multiple epitopes and ability to bind several HLA class I and class II molecules for presentation to cells of the cellular immune response, may be useful as a peptide-based vaccine against tuberculosis.

Keywords: Mycobacterium tuberculosis, Rv3873, peptides, vaccine

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2 Achievements of Healthcare Services Vis-À-Vis the Millennium Development Goals Targets: Evidence from Pakistan

Authors: Saeeda Batool, Ather Maqsood Ahmed

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This study investigates the impact of public healthcare facilities and socio-economic circumstances on the status of child health in Pakistan. The complete analysis is carried out in correspondence with fourth and sixth millennium development goals. Further, the health variables chosen are also inherited from targeted indicators of the mentioned goals (MDGs). Trends in the Human Opportunity Index (HOI) for both health inequalities and coverage are analyzed using the Pakistan Social and Living Standards Measurement (PLSM) data set for 2001-02 to 2012-13 at the national and provincial level. To reveal the relative importance of each circumstance in achieving the targeted values for child health, Shorrocks decomposition is applied on HOI. The annual point average growth rate of HOI is used to simulate the time period for the achievement of target set by MDGs and universal access also. The results indicate an improvement in HOI for a reduction in child mortality rates from 52.1% in 2001-02 to 67.3% in 2012-13, which confirms the availability of healthcare opportunities to a larger segment of society. Similarly, immunization against measles and other diseases such as Diphtheria, Polio, Bacillus Calmette-Guerin (BCG), and Hepatitis has also registered an improvement from 51.6% to 69.9% during the period of study at the national level. On a positive note, no gender disparity has been found for child health indicators and that health outcome is mostly affected by the parental and geographical features and availability of health infrastructure. However, the study finds that this achievement has been uneven across provinces. Pakistan is not only lagging behind in achieving its health goals, disappointingly with the current rate of health care provision, but it will take many additional years to achieve its targets.

Keywords: Socio-economic circumstances, unmet MDGs, public healthcare services, child and infant mortality.

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1 The Evaluation of Antioxidant and Antimicrobial Activities of Essential Oil and Aqueous, Methanol, Ethanol, Ethyl Acetate and Acetone Extract of Hypericum scabrum

Authors: A. Heshmati, M. Y Alikhani, M. T. Godarzi, M. R. Sadeghimanesh

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Herbal essential oil and extracts are a good source of natural antioxidants and antimicrobial compounds. Hypericum is one of the potential sources of these compounds. In this study, the antioxidant and antimicrobial activity of essential oil and aqueous, methanol, ethanol, ethyl acetate and acetone extract of Hypericum scabrum was assessed. Flowers of Hypericum scabrum were collected from the surrounding mountains of Hamadan province and after drying in the shade, the essential oil of the plant was extracted by Clevenger and water, methanol, ethanol, ethyl acetate and acetone extract was obtained by maceration method. Essential oil compounds were identified using the GC-Mass. The Folin-Ciocalteau and aluminum chloride (AlCl3) colorimetric method was used to measure the amount of phenolic acid and flavonoids, respectively. Antioxidant activity was evaluated using DPPH and FRAP. The minimum inhibitory concentration (MIC) and the minimum bacterial/fungicide concentration (MBC/MFC) of essential oil and extracts were evaluated against Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella typhimurium, Aspergillus flavus and Candida albicans. The essential oil yield of was 0.35%, the lowest and highest extract yield was related to ethyl acetate and water extract. The most component of essential oil was α-Pinene (46.35%). The methanol extracts had the highest phenolic acid (95.65 ± 4.72 µg galic acid equivalent/g dry plant) and flavonoids (25.39 ± 2.73 µg quercetin equivalent/g dry plant). The percentage of DPPH radical inhibition showed positive correlation with concentrations of essential oil or extract. The methanol and ethanol extract had the highest DDPH radical inhibitory. Essential oil and extracts of Hypericum had antimicrobial activity against the microorganisms studied in this research. The MIC and MBC values for essential oils were in the range of 25-25.6 and 25-50 μg/mL, respectively. For the extracts, these values were 1.5625-100 and 3.125-100 μg/mL, respectively. Methanol extracts had the highest antimicrobial activity. Essential oil and extract of Hypericum scabrum, especially methanol extract, have proper antimicrobial and antioxidant activity, and it can be used to control the oxidation and inhibit the growth of pathogenic and spoilage microorganisms. In addition, it can be used as a substitute for synthetic antioxidant and antimicrobial compounds.

Keywords: Antimicrobial, antioxidant, extract, hypericum.

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