Search results for: strains

Authors: L. S. Markosyan, L. A. Ganushkina, N. S. Vardanyan, K. V. Harutyunova, M. V. Harutyunova

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Five original strains of entomopathogenic bacteria with insecticidal activity against mosquito larvae of the genera Aedes, Culex and Anopheles have been isolated from natural conditions in Armenia and characterized. According to morphological, physiological and biochemical parameters, all isolates were identified as Bacillus thuringiensis spp. israelensis (Bti). High larvicidal activity has been showed by three strains Bti. These strains can be recommended for industrial production of bacterial preparations.

Keywords: Armenia, Bacillus thuringiensis ssp. israelensis, bloodsucking mosquito control, new effective strains of bacteria.

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Authors: S. A. Aytkeldiyeva, A. K. Sadanov, E. R. Faizulina, A. A. Kurmanbayev

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15 strains of oil-destructing microorganisms were isolated from oil polluted soil of Western Kazakhstan. Strains 2-A and 41-3 with the highest oil-destructing activities were chosen from them. It was shown that these strains oxidized n-alkanes very well, but isoalkanes, isoparaffin, cycloparaffin and heavy aromatic compounds were destructed very slowly. These both strains were tested as preparations for bioremediation of oil-polluted soil in model and field experiments. The degree of utilizing of soil oil by this preparation was 79-84 % in field experiments.

Keywords: Bioremediation, n-alkanes, oil-polluted soil, oiloxidizingmicroorganisms.

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Authors: Amr A. El Hanafy, Yasir Anwar, Saleh A. Mohamed, Saleh Mohamed Saleh Al-Garni, Jamal S. M. Sabir, Osama A. H. Abu Zinadah, Mohamed Morsi Ahmed

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In the vicinity of red sea about 15 fungi species were isolated from oil contaminated sites. On the basis of aptitude to degrade the crude oil and DCPIP assay, two fungal isolates were selected amongst 15 oil degrading strains. Analysis of ITS-1, ITS-2 and amplicon pyrosequencing studies of fungal diversity revealed that these strains belong to Penicillium and Aspergillus species. Two strains that proved to be the most efficient in degrading crude oil was Aspergillus niger (54%) and Penicillium commune (48%) Subsequent to two weeks of cultivation in BHS medium the degradation rate were recorded by using spectrophotometer and GC-MS. Hence, it is cleared that these fungal strains has capability of degradation and can be utilize for cleaning the Saudi Arabian environment.

Keywords: Fungal strains, hydrocarbon contaminants, molecular identification, biodegradation, GC-MS.

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Authors: A. Kistaubayeva, I. Savitskaya, D. Ibrayeva, M. Abdulzhanova, N. Voronova

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36 strains of sulfur-oxidizing bacteria were isolated in Southern Kazakhstan soda-saline soils and identified. Screening of strains according bio-oxidation (destruction thiosulfate to sulfate) and enzymatic (Thiosulfate dehydrogenises and thiosulfate reductase) activity was conducted. There were selected modes of aeration and culture conditions (pH, temperature), which provide optimum harvest cells. These strains can be used in bio-melioration technology.

Keywords: Elemental sulfur, oxidation activity, Тhiobacilli, fertilizers, heterotrophic S-oxidizers.

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Authors: Ming-Lun Chiang, Hsi-Chia Chen, Chieh Wu, Yu-Ting Tseng, Ming-Ju Chen

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In this study, three strains of Vibrio parahaemolyticus (690, BCRC 13023 and BCRC 13025) were subjected to acid adaptation at pH 5.5 for 90 min. The survival of acid-adapted and non-adapted V. parahaemolyticus strains under simulated gastric condition and their protein expression profiles were investigated. Results showed that acid adaptation increased the survival of the test V. parahaemolyticus strains after exposure to simulated gastric juice (pH 3). Additionally, acid adaptation also affected the protein expression in these V. parahaemolyticus strains. Nine proteins, identified as atpA, atpB, DnaK, GroEL, OmpU, enolase, fructose-bisphosphate aldolase, phosphoglycerate kinase and triosephosphate isomerase, were induced by acid adaptation in two or three of the test strains. These acid-adaptive proteins may play important regulatory roles in the acid tolerance response (ATR) of V. parahaemolyticus.

Keywords: Acid adaptation, protein expression, simulated gastric juice, Vibrio parahaemolyticus

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Authors: Gomaa N. Abdel-Rahman, Nadia R. A. Nassar, Yehia A. Heikal, Mahmoud A. M. Abou-Donia, Mohamed B. M. Ahmed, Mohamed Fadel

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Storage stability is the important factor of baker's yeast quality. Effect of the storage period (fifteen days) on storage sugars and cell viability of baker's yeast, produced from three S. cerevisiae strains (FC-620, FH-620, and FAT-12) as comparison with baker's yeast produced by S. cerevisae F-707 (original strain of baker's yeast factory) were investigated. Studied trehalose and glycogen content ranged from 10.19 to 14.79 % and from 10.05 to 10.69 % (d.w.), respectively before storage. The trehalose and glycogen content of all strains was decreased by increasing the storage period with no significant differences between the reduction rates of trehalose. Meanwhile, reduction rates of glycogen had significant differences between different strains, where the FH-620 and FC-620 strains had lowest rates as 18.12 and 20.70 %, respectively. Also, total viable cells and gassing power of all strains were decreased by increasing the storage period. FH-620 and FC-620 strains had the lowest values of reduction rates as an indicator of storage resistant. Where the reduction rates in total viable cells of FH-620 and FC-620 strains were 22.05 and 24.70%, respectively, while the reduction rates of gassing power were 20.90 and 24.30%, in the same order. On other hand, FAT-12 strain was more sensitive to storage as compared to original strain, where the reduction rates were 35.60 and 35.75%, respectively for total viable cells and gassing power.

Keywords: Baker’s yeast, trehalose, glycogen, gassing power.

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Authors: M. Raissy, M. Shahrani

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The present study was done to evaluate the presence of tetracycline resistance genes in Lactococcus garvieae isolated from cultured rainbow trout, West Iran. The isolates were examined for antimicrobial resistance using disc diffusion method. Of the 49 strains tested, 19 were resistant to tetracycline (38.7%), 32 to enrofloxacin (65.3%), 21 to erythromycin (42.8%), 20 to chloramphenicol and trimetoprim-sulfamethoxazole (40.8%). The strains were then characterized for their genotypic resistance profiles. The results revealed that all 49 isolates contained at least one of the tetracycline resistance genes. Tet (A) was found in 89.4% of tetracycline resistant isolates and the frequency of other gene were as follows: tet (E) 42.1%, tet (B) 47.3%, tet (D) 15.7%, tet (L) 26.3%, tet (K) 52.6%, tet (G) 36.8%, tet (34) 21%, tet (S) 63.1%, tet (C) 57.8%, tet (M) 73.6%, tet (O) 42.1%. The results revealed high levels of antibiotic resistance in L. garvieae strains which is a potential danger for trout culture as well as for public health.

Keywords: Lactococcus garvieae, rainbow trout, tetracycline resistance genes.

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Authors: A. Simoneau

Abstract:

Metal cutting is a severe plastic deformation process involving large strains, high strain rates, and high temperatures. Conventional analysis of the chip formation process is based on bulk material deformation disregarding the inhomogeneous nature of the material microstructure. A series of orthogonal cutting tests of AISI 1045 and 1144 steel were conducted which yielded similar process characteristics and chip formations. With similar shear angles and cut chip thicknesses, shear strains for both chips were found to range from 2.0 up to 2.8. The manganese-sulfide (MnS) precipitate in the 1144 steel has a very distinct and uniform shape which allows for comparison before and after chip formation. From close observations of MnS precipitates in the cut chips it is shown that the conventional approach underestimates plastic strains in metal cutting. Experimental findings revealed local shear strains around a value of 6. These findings and their implications are presented and discussed.

Keywords: Machining, metal cutting, microstructure, plastic strains, local strain.

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Authors: Thanh-Duoc Phan, Saden H. Zahiri, S. H. Masood, Mahnaz Jahedi

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In this article, using finite element analysis (FEA) and an X-ray diffractometer (XRD), cold-sprayed titanium particles on a steel substrate is investigated in term of cooling time and the development of residual strains. Three cooling-down models of sprayed particles after deposition stage are simulated and discussed: the first model (m1) considers conduction effect to the substrate only, the second model (m2) considers both conduction as well as convection effect to the environment, and the third model (m3) which is the same as the second model but with the substrate heated to a near particle temperature before spraying. Thereafter, residual strains developed in the third model is compared with the experimental measurement of residual strains, which involved a Bruker D8 Advance Diffractometer using CuKa radiation (40kV, 40mA) monochromatised with a graphite sample monochromator. For deposition conditions of this study, a good correlation was found to exist between the FEA results and XRD measurements of residual strains.

Keywords: cold gas dynamic spray, X-ray diffraction, explicit finite element analysis, residual strain, titanium, particle impact, deformation behavior.

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Authors: Ming-Lun Chiang, Chieh Wu, Ming-Ju Chen

Abstract:

Three strains of Vibrio parahaemolyticus (690, BCRC 13023 and BCRC 13025) implicated in food poisoning outbreaks in Taiwan were subjected to acid adaptation at pH 5.5 for 90 min. The growth behaviors of acid-adapted and non-adapted V. parahaemolyticus in the media supplemented with various nitrogen and carbon sources were investigated. The effects of acid adaptation on the thermostable direct hemolysin (TDH) secretion and fatty acid profiles of V. parahaemolyticus were also examined. Results showed that acid-adapted and non-adapted V. parahaemolyticus 690, BCRC 13023 and BCRC 13025 grew similarly in TSB-3% NaCl and basal media supplemented with various carbon and nitrogen sources during incubation period. Higher TDH secretion was noted with V. parahaemolyticus 690 among the three strains. However, acid-adapted strains produced less amounts of TDH than non-adapted strains when they were grown in TSB-3% NaCl. Additionally, acid adaptation increased the ratio of SFA/USFA in cells of V. parahaemolyticus strains.

Keywords: Vibrio parahaemolyticus, acid adaptation, thermostable direct hemolysin, fatty acid profile.

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Authors: Nadia R. A. Nassar, Yehia A. Heikal, Mahmoud A. M. Abou Donia, Mohamed Fadel, Gomaa N. Abdel-Rahman

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The aim of this study was to select the best strains of Saccharomyces cerevisiae able to resist lead and cadmium. Ten strains were screened on the basis of their resistance at different concentrations of 0, 2, 4, 8, 12, 16, 20 and 24 ppm for Pb and 0, 0.5, 1, 2, 4, 6, 8 and 10 ppm for Cd. The properties of baker's yeast quality were decreased by the increase of Pb or Cd in growth medium. The slope values of yield, total viable cells and gassing power of produced baker's yeast were investigated as an indicator of metal resistant. In addition, concentrations of Pb and Cd in produced baker's yeast were determined. The strain of S. cerevisiae FH-620 had the highest resistance against Pb and Cd and had the minimum levels of both two investigated metals in produced baker's yeast.

Keywords: Cadmium, lead, S. cerevisiae, tolerant.

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Authors: Siti Aishah Abu Bakar, Madihah Zawawi, Abdul Manaf Ali, Aini Ideris

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Newcastle Disease Virus (NDV), an avian paramyxovirus, is a highly contagious, generalised virus disease of domestic poultry and wild birds characterized by gastro-intestinal, respiratory and nervous signs. In this study, it was shown that NDV strain AF2240 and V4-UPM are cytolytic to Human Promyelocytic Leukemia, HL60 and Human T-lymphoblastic Leukemia, CEM-SS cells. Results from MTT cytolytic assay showed that CD50 for NDV AF2240 against HL60 was 130 HAU and NDV V4-UPM against HL60 and CEM-SS were 110.6 and 150.9 HAU respectively. Besides, both strains were found to inhibit the proliferation of cells in a dose dependent manner. The mode of cell death either by apoptosis or necrosis was further analyzed using acridine orange and propidium iodide (AO/PI) staining. Our results showed that both NDV strains induced primarily apoptosis in treated cells at CD50 concentration. In conclusion, both NDV strains caused cytolytic effects primarily via apoptosis in leukemia cells.

Keywords: Apoptosis, Cytolytic, Leukaemia, Newcastle DiseaseVirus

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Authors: Natalia Koltovaya, Nadezhda Zhuchkina, Ksenia Lyubimova

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We study the biological effects induced by ionizing radiation in view of therapeutic exposure and the idea of space flights beyond Earth's magnetosphere. In particular, we examine the differences between base pair substitution induction by ionizing radiation in model haploid and diploid yeast Saccharomyces cerevisiae cells. Such mutations are difficult to study in higher eukaryotic systems. In our research, we have used a collection of six isogenic trp5-strains and 14 isogenic haploid and diploid cyc1-strains that are specific markers of all possible base-pair substitutions. These strains differ from each other only in single base substitutions within codon-50 of the trp5 gene or codon-22 of the cyc1 gene. Different mutation spectra for two different haploid genetic trp5- and cyc1-assays and different mutation spectra for the same genetic cyc1-system in cells with different ploidy — haploid and diploid — have been obtained. It was linear function for dose-dependence in haploid and exponential in diploid cells. We suggest that the differences between haploid yeast strains reflect the dependence on the sequence context, while the differences between haploid and diploid strains reflect the different molecular mechanisms of mutations.

Keywords: Base pair substitutions, γ-rays, haploid and diploid cells, yeast Saccharomyces cerevisiae.

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Authors: A. J. Sanda, O. Olowofeso, M. A. Adeleke, A. O Oso, S. O. Durosaro, M. O. Sanda

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A total of 150 meat type chickens comprising 50 each of Arbor Acre, Marshall and Ross were used for this study which lasted for 10 weeks at the Federal University of Agriculture, Abeokuta, Nigeria. Growth performance data were collected from the third week through week 10 and data obtained were analysed using the Generalized Linear Model Procedure. Heritability estimates (h2) for body dimensions carried out on the chicken strains ranged from low to high. Marshall broiler chicken strain had the highest h2 for body weight 0.46±0.04, followed by Arbor Acre and Ross with h2 being 0.38±0.12 and 0.26±0.06, respectively. The repeatability estimates for body weight in the three broiler strains were high, and it ranged from 0.70 at week 4 to 0.88 at week 10. Relationships between the body weight and linear body measurements in the broiler chicken strains were positive and highly significant (p > 0.05).

Keywords: Broiler chicken strains, heritability, repeatability, traits.

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Authors: Ali Zazoua, Anis Zazoua, Ahcen Taleb, Nicole Jaffrezic-Renault

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Although oil-based drilling fluids are of paramount practical and economical interest, they represent a serious source of pollution, once released into the environment as drill cuttings. The aim of this study is to assess the capability of isolated microorganisms to degrade gasoil fuel. The commonly used physicochemical and biodegradation remediation techniques of petroleum contaminated soil were both investigated. The study revealed that natural biodegradation is favorable. Even though, the presence of heavy metals, the moisture level of (8.55%) and nutrient deficiencies put severe constrains on microorganisms- survival ranges inhibiting the biodegradation process. The selected strains were able to degrade the diesel fuel at significantly high rates (around 98%).

Keywords: Biodegradation, Gasoil, Pollution, Microbial strains, Hydrocarbon, soil pollution

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Authors: S. Liepina, A. Jemeljanovs

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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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Authors: Nga Phuong Dang, Terje Vasskog, Ashwiny Pandey, Rajnish Kaur Calay

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Diatoms hold great potential for nutraceutical production as they are source of high value products such as eicosapentaenoic acid (EPA) and pigment fucoxanthin. EPA has proven human health benefits, and fucoxanthin can be used as both medicinal and nutritional ingredient to prevent and treat chronic diseases. The economic perspective of commercial production of a single product from microalgae is not attractive due to the high production cost. To improve the economic viability, we explore the concept of combining the production of both EPA and fucoxanthin in a single process. In our current study, we isolated twelve new microalgae isolates from Ofotfjord. Eight of them are diatoms and 4 of them are cyanobacteria and microalgae. Screening the new diatom isolates revealed that two strains are cold-tolerant diatom which can grow at 10 °C. They accumulated significant amount of lipid, which was up to 40-60% of the dried mass. The EPA contents from the two strains ranged from 15-19% of the total fatty acid, while fucoxanthin concentrations were between 1-1.4% of the dried biomass. Comparing with other studied diatom, this is promising result. The two strains hold promise as source for EPA and fucoxanthin production.

Keywords: Microalgae, fucoxanthin, eicosapentaenoic acid, diatom, fatty acid.

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Authors: Daomao Yang, Qizhi Wang

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Monoterpenoids are mainly found in plant essential oils and they are ideal substrates for biotransformation into oxygen-containing derivatives with important commercial value due to their low price and simple structure. In this paper, eleven strains of endophytic fungi from Praxelis clematidea were used as test strains to conduct the whole cell biotransformation of the monoterpenoids: (+)-limonene, (-)-limonene and myrcene. The fungi were inoculated in 50 ml Sabouraud medium and incubated at 30 ℃ with the agitation of 150 r/min for 6 d, and then 0.5% (v/v) substrates were added into the medium and biotransformed for further 3 d. Afterwards the cultures were filtered, and extracted using equal volume of ethyl acetate. The metabolites were analyzed by GC-MS technique with NIST database. The Total Ion Chromatogram of the extractions from the eleven strains showed that the main product of (+)- and (-)-limonene biotransformation was limonene-1,2-diol, while it is limonene and linalool oxide for biotransformation of myrcene. This work will help screen the microorganisms to biotransform the monoterpenes.

Keywords: Endophytic fungi, (+)–limonene, (-)–limonene, myrcene.

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Authors: Fereshteh Shacheraghi, Mohammad Reza Shakibaie, Hanieh Noveiri

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Fourty one strains of ESBL producing P.aeruginosa which were previously isolated from burn patients in Kerman University general hospital, Iran were subjected to PCR, RFLP and sequencing in order to determine the type of extended spectrum β- lactamases (ESBL), the restriction digestion pattern and possibility of mutation among detected genes. DNA extraction was carried out by phenol chloroform method. PCR for detection of bla genes was performed using specific primer for each gene. Restriction Fragment Length Polymorphism (RFLP) for ESBL genes was carried out using EcoRI, NheI, PVUII, EcoRV, DdeI, and PstI restriction enzymes. The PCR products were subjected to direct sequencing of both the strands for identification of the ESBL genes.The blaCTX-M, blaVEB-1, blaPER-1, blaGES-1, blaOXA-1, blaOXA-4 and blaOXA-10 genes were detected in the (n=1) 2.43%, (n=41)100%, (n=28) 68.3%, (n=10) 24.4%, (n=29) 70.7%, (n=7)17.1% and (n=38) 92.7% of the ESBL producing isolates respectively. The RFLP analysis showed that each ESBL gene has identical pattern of digestion among the isolated strains. Sequencing of the ESBL genes confirmed the genuinety of PCR products and revealed no mutation in the restriction sites of the above genes. From results of the present investigation it can be concluded that blaVEB-1 and blaCTX-M were the most and the least frequently isolated ESBL genes among the P.aeruginosa strains isolated from burn patients. The RFLP and sequencing analysis revealed that same clone of the bla genes were indeed existed among the antibiotic resistant strains.

Keywords: ESBL genes, PCR, RFLP, Sequencing, P.aeruginosa

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Authors: Eleftheria Tzamali, Panayiota Poirazi, Ioannis G. Tollis, Martin Reczko

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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.

Keywords: Bacterial polymorphism, clique identification, dynamic FBA, evolution, metabolic interactions.

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Authors: I. Savitskaya, A. Kistaubayeva, A. Zhubanova, I. Blavachinskaiya, D. Ibrayeva, M. Abdulzhanova, A. Otarbay, A.Isabekova

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This study was conducted for the investigation of number of cellulolytic bacteria and their ability in decomposition. Seven samples surface soil were collected on cellulose Zailiskii Alatau slopes. Cellulolitic activity of new strains of Bacillus, isolated from soil is determined. Isolated cellulose degrading bacteria were screened for determination of the highest cellulose activity by quantitative assay using Congo red, gravimetric assay and colorimetric DNS method trough of the determination of the parameters of sugar reduction. Strains are assigned to: B.subtilis, B.licheniformis, B. cereus and, В. megaterium. Bacillus strains consisting of several different types of cellulases have broad substrate specificity of cellulase complexes formed by them. Cellulolitic bacteria were recorded to have highest cellulase activity and selected for optimization of cellulase enzyme production.

Keywords: Cellulose-degrading bacteria, cellulase complex, foothills soil, screening.

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Authors: Jatindra N. Bhakta, Kouhei Ohnishi, Yukihiro Munekage, Kozo Iwasaki

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The growing health hazardous impact of arsenic (As) contamination in environment is the impetus of the present investigation. Application of lactic acid bacteria (LAB) for the removal of toxic and heavy metals from water has been reported. This study was performed in order to isolate and characterize the Asresistant LAB from mud and sludge samples for using as efficient As uptaking probiotic. Isolation of As-resistant LAB colonies was performed by spread plate technique using bromocresol purple impregnated-MRS (BP-MRS) agar media provided with As @ 50 μg/ml. Isolated LAB were employed for probiotic characterization process, acid and bile tolerance, lactic acid production, antibacterial activity and antibiotic tolerance assays. After As-resistant and removal characterizations, the LAB were identified using 16S rDNA sequencing. A total of 103 isolates were identified as As-resistant strains of LAB. The survival of 6 strains (As99-1, As100-2, As101-3, As102-4, As105-7, and As112-9) was found after passing through the sequential probiotic characterizations. Resistant pattern pronounced hollow zones at As concentration >2000 μg/ml in As99-1, As100-2, and As101-3 LAB strains, whereas it was found at ~1000 μg/ml in rest 3 strains. Among 6 strains, the As uptake efficiency of As102-4 (0.006 μg/h/mg wet weight of cell) was higher (17 – 209%) compared to remaining LAB. 16S rDNA sequencing data of 3 (As99- 1, As100-2, and As101-3) and 3 (As102-4, As105-7, and As112-9) LAB strains clearly showed 97 to 99% (340 bp) homology to Pediococcus dextrinicus and Pediococcus acidilactici, respectively. Though, there was no correlation between the metal resistant and removal efficiency of LAB examined but identified elevated As removing LAB would probably be a potential As uptaking probiotic agent. Since present experiment concerned with only As removal from pure water, As removal and removal mechanism in natural condition of intestinal milieu should be assessed in future studies.

Keywords: Lactic acid bacteria, As-resistant, characterization, Pediococcus sp., As removal probiotic.

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Authors: Ngo Thanh Xuan, Mai Thi Hang, Vu Nguyen Thanh

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A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.

Keywords: Aspergillus niger XP, cloning, over expression, Pichia pastoris, phyA , phytase.

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Authors: T. Buyuksirit, H. Kuleasan

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Natural antimicrobials are used to preserve foods that can be found in plants, animals, and microorganisms. Antimicrobial substances are natural or artificial agents that produced by microorganisms or obtained semi/total chemical synthesis are used at low concentrations to inhibit the growth of other microorganisms. Food borne pathogens and spoilage microorganisms are inactivated by the use of antagonistic microorganisms and their metabolites. Yeasts can produce toxic proteins or glycoproteins (toxins) that cause inhibition of sensitive bacteria and yeast species. Antimicrobial substance producing phenotypes belonging different yeast genus were isolated from different sources. Toxins secreted by many yeast strains inhibiting the growth of other yeast strains. These strains show antimicrobial activity, inhibiting the growth of mold and bacteria. The effect of antimicrobial agents produced by yeasts can be extremely fast, and therefore may be used in various treatment procedures. Rapid inhibition of microorganisms is possibly caused by microbial cell membrane lipopolysaccharide binding and in activation (neutralization) effect. Antimicrobial agents inhibit the target cells via different mechanisms of action.

Keywords: Antimicrobial agents, Glycoprotein, Toxic protein, Yeast.

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Authors: A. El Behiry, R. N. Zahran, R. Tarabees, E. Marzouk, M. Al-Dubaib

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Mastitis is one of the most economic disease affecting dairy cows worldwide. Its classic diagnosis using bacterial culture and biochemical findings is a difficult and prolonged method. In this research, using of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) permitted identification of different microorganisms with high accuracy and rapidity (only 24 hours for microbial growth and analysis). During the application of MALDI-TOF MS, one hundred twenty strains of Staphylococcus and Streptococcus species isolated from milk of cows affected by clinical and subclinical mastitis were identified, and the results were compared with those obtained by traditional methods as API and VITEK 2 Systems. 37 of totality 39 strains (~95%) of Staphylococcus aureus (S. aureus) were exactly detected by MALDI TOF MS and then confirmed by a nuc-based PCR technique, whereas accurate identification was observed in 100% (50 isolates) of the coagulase negative staphylococci (CNS) and Streptococcus agalactiae (31 isolates). In brief, our results demonstrated that MALDI-TOF MS is a fast and truthful technique which has the capability to replace conventional identification of several bacterial strains usually isolated in clinical laboratories of microbiology.

Keywords: Identification, mastitis pathogens, mass spectral, phenotypical.

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Authors: M. Mahboubkhah, H. Fayazfar

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In this research, Forming Limit Diagrams for supertension sheet metals which are using in automobile industry have been obtained. The exerted strains to sheet metals have been measured with four different methods and the errors of each method have also been represented. These methods have been compared with together and the most efficient and economic way of extracting of the exerted strains to sheet metals has been introduced. In this paper total error and uncertainty of FLD extraction procedures have been derived. Determination of the measurement uncertainty in extracting of FLD has a great importance in design and analysis of the sheet metal forming process.

Keywords: Forming Limit Diagram, Major and Minor Strain, Measurement Uncertainty.

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Authors: E. Soboleva, E. Sergachyova, S. G. Davydenko, T. V. Meledina

Abstract:

Problem of food preservation is extremely important for mankind. Viscous damage ("illness") of bread results from development of Bacillus spp. bacteria. High temperature resistant spores of this microorganism are steady against 120°C) and remain in bread during pastries, potentially causing spoilage of the final product. Scientists are interested in further characterization of bread spoiling Bacillus spp. species. Our aim was to find weather yeast Saccharomyces cerevisiae strains that are able to produce natural antimicrobial killer factor can preserve bread illness. By diffusion method, we showed yeast antagonistic activity against spore-forming bacteria. Experimental technological parameters were the same as for bakers' yeasts production on the industrial scale. Risograph test during dough fermentation demonstrated gas production. The major finding of the study was a clear indication of the presence of killer yeast strain antagonistic activity against rope in bread causing bacteria. After demonstrating antagonistic effect of S. cerevisiae on bacteria using solid nutrient medium, we tested baked bread under provocative conditions. We also measured formation of carbon dioxide in the dough, dough-making duration and quality of the final products, when using different strains of S. cerevisiae. It is determined that the use of yeast S. cerevisiae RCAM 01730 killer strain inhibits appearance of rope in bread. Thus, natural yeast antimicrobial killer toxin, produced by some S. cerevisiae strains is an anti-rope in bread protector.

Keywords: Bakers' yeasts, rope in bread, Saccharomyces cerevisiae.

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Authors: Sarma. M.V.R.K, Saharan. K, Prakash. A, Bisaria. V. S, Sahai.V

Abstract:

Vermiculite was used to develop inorganic carrier-based formulations of fluorescent pseudomonad strains R62 and R81. The effect of bio-inoculation of fluorescent pseudomonad strains R62 and R81 (plant growth promoting and biocontrol agent) on growth responses of Vigna-mungo under field condition was enumerated. The combined bioinoculation of these two organisms in a formuation increased the pods yield by 300% in comparison to the control crop. There was also significant increment in the other plant growth responses such as dry root weight, dry shoot weight, shoot length and number of branches per plant.

Keywords: Bio-inoculants formulation, Fluorescent pseudomonad, Plant growth promotion activity.

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Authors: Mihriban Korukluoglu, Goksen Arik, Cagla Erdogan, Selen Kocakoglu

Abstract:

Kefir is a traditional fermented refreshing beverage which is known for its valuable and beneficial properties for human health. Mainly yeast species, lactic acid bacteria (LAB) strains and fewer acetic acid bacteria strains live together in a natural matrix named “kefir grain”, which is formed from various proteins and polysaccharides. Different microbial species live together in slimy kefir grain and it has been thought that synergetic effect could take place between microorganisms, which belong to different genera and species. In this research, yeast and LAB were isolated from kefir samples obtained from Uludag University Food Engineering Department. The cell morphology of isolates was screened by microscopic examination. Gram reactions of bacteria isolates were determined by Gram staining method, and as well catalase activity was examined. After observing the microscopic/morphological and physical, enzymatic properties of all isolates, they were divided into the groups as LAB and/or yeast according to their physicochemical responses to the applied examinations. As part of this research, the antagonistic/synergistic efficacy of the identified five LAB and five yeast strains to each other were determined individually by disk diffusion method. The antagonistic or synergistic effect is one of the most important properties in a co-culture system that different microorganisms are living together. The synergistic effect should be promoted, whereas the antagonistic effect is prevented to provide effective culture for fermentation of kefir. The aim of this study was to determine microbial interactions between identified yeast and LAB strains, and whether their effect is antagonistic or synergistic. Thus, if there is a strain which inhibits or retards the growth of other strains found in Kefir microflora, this circumstance shows the presence of antagonistic effect in the medium. Such negative influence should be prevented, whereas the microorganisms which have synergistic effect on each other should be promoted by combining them in kefir grain. Standardisation is the most desired property for industrial production. Each microorganism found in the microbial flora of a kefir grain should be identified individually. The members of the microbial community found in the glue-like kefir grain may be redesigned as a starter culture regarding efficacy of each microorganism to another in kefir processing. The main aim of this research was to shed light on more effective production of kefir grain and to contribute a standardisation of kefir processing in the food industry.

Keywords: Antagonistic effect, kefir, lactic acid bacteria (LAB), synergistic, yeast.

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Authors: Ahmed F. Azmy , Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed

Abstract:

Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosa (PS1), and Pseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazinon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 10^12CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Keywords: Acinetobacter baumannii, biodegradation, Malathion, organophosphate pesticides.

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