Search results for: Recombinant E.coli

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

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

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

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

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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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Authors: Phanwipa Pangsri, Yawariyah Weahayee

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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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Authors: Imana Shahrin Tania, Mohammad Ali

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

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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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Authors: Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol Karim Sheikhi, Mehdi Hedayati

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Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.

Keywords: hTSH, bioinformatics, protein expression, cross reactivity.

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Authors: Archana Panchapakesan Iyer, Taha Abdullah Kumosani

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

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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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Authors: Simona Dostalova, Ana Maria Jimenez Jimenez, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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Various nanomaterials can be used as a drug delivery vehicles in nanomedicine, called nanocarriers. They can either be organic or inorganic, synthetic or natural-based. Although synthetic nanocarriers are easier to produce, they can often be toxic for the organism and thus not suitable for use in treatment. From naturalbased nanocarriers, the most commonly used are protein cages or viral capsids. In this work, virus bacteriophage λ was used for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2% maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000×g and 4°C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 μg·mL-1). Free drug molecules were removed by filtration. The encapsulation was verified using the absorbance for doxorubicin and atomic absorption spectrometry for platinum cytostatics. The amount of encapsulated drug linearly increased with the increasing concentration of applied drug with the determination coefficient R2=0.989 for doxorubicin; R2=0.967 for cisplatin; R2=0.989 for carboplatin and R2=0.996 for oxaliplatin. The overall encapsulation efficiency was calculated as 50% for doxorubicin; 8% for cisplatin; 6% for carboplatin and 10% for oxaliplatin.

Keywords: Bacteriophage λ, doxorubicin, platinum cytostatics, protein-based nanocarrier, viral capsid.

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Authors: S. Angalaparameswari, T.S. Mohamed Saleem, M. Alagusundaram, S. Ramkanth, V.S. Thiruvengadarajan, K. Gnanaprakash, C. Madhusudhana Chetty, G. Pratheesh

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The present research was designed to investigate the anti-microbial activity of aristolochic acid from the root of Aristolochia bracteata. From the methanolic & ethyl extract extracts of Aristolochia bracteata aristolochic acid I was isolated and conformed through IR, NMR & MS. The percentage purity of aristolochic acid I was determined by UV & HPLC method. Antibacterial activity of extracts of Aristolochia bracteata and the isolated compound was determined by disc diffusion method. The results reveled that the isolated aristolochic acid from methanolic extract was more pure than the compound from ethyl acetate extract. The various extracts (500μg/disc) of Aristolochia bracteata showed moderate antibacterial activity with the average zone of inhibition of 7-18 mm by disc diffusion method. Among the extracts, ethyl acetate & methanol extracts were shown good anti-microbial activity and the growth of E.coli (18 mm) was strongly inhibited. Microbial assay of isolated compound (Aristolochic acid I) from ethyl acetate & methanol extracts were shown good antimicrobial activity and the zone of inhibition of both at higher concentration 50 μg/ml was similar with the standard aristolochic acid. It may be concluded that the isolated compound of aristolochic acid I has good anti-bacterial activity.

Keywords: Aristolochic acid I, Anti-microbial activity, Aristolochia bracteata, Bacillus subtilis, E.coli

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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: Samy A. Selim

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The antimicrobial, antiplasmid and cytotoxic activities of marine algae Halimeda opuntia and Sarconema filiforme were investigated. Antimicrobial bioassay against some human pathogenic bacteria and yeast were conducted using disc diffusion method. Halimeda extract exhibited antibacterial activity against six species of microrganisms, with significant inhibition against Staphylococcus aureus. While Sarconema extract was better potent as antifungal against Candida albicans. Comparative antibacterial studies showed that Halimeda extract showed equivalent or better activity as compared with commercial antibiotic when tested against Staphylococcus aureus. Further tests conducted using dilution method showed both extracts as having bacteriostatic mode of action against the tested microorganisms. Methanol extract of two species showed significant cytotoxicity (LC50 <500μg) on brine shrimp. Halimeda opuntia showed highest cytotoxic activity (LC50 =192.3μg). Also, the present investigation was undertaken to investigate the ability of methanolic extract of the algal extracts to cure R-plasmids from certain clinical E. coli isolates. The active fraction of Halimeda and Sarconema could cure plasmids from E. coli at curing efficiencies of approximately 78%. The active fraction mediated plasmid curing resulted in the subsequent loss of antibiotic resistance encoded in the plasmids as revealed by antibiotic resistance profile of cured strains. The screening results confirm the possible use of marine algae Halimeda opuntia and Sarconema filiforme as a source of pharmacological benefits.

Keywords: Antimicrobial, antiplasmid Cytotoxicity, Marine Algae.

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Authors: Wai Prathumpai, Pranee Rachtawee, Sutamat Khajeeram, Pariya Na Nakorn

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The  β-glucan produced by Ophiocordyceps dipterigena BCC 2073 is a (1, 3)-β-D-glucan with highly branching O-6-linkedside chains that is resistant to acid hydrolysis (by hydrochloric acid and porcine pancreatic alpha-amylase). This β-glucan can be utilized as a prebiotic due to its advantageous structural and biological properties. The effects of using this β-glucan as the sole carbon source for the in vitro growth of two probiotic bacteria (L. acidophilus BCC 13938 and B. animalis ATCC 25527) were investigated. Compared with the effect of using 1% glucose or fructo-oligosaccharide (FOS) as the sole carbon source, using 1% β-glucan for this purpose showed that this prebiotic supported and stimulated the growth of both types of probiotic bacteria and induced them to produce the highest levels of metabolites during their growth. The highest levels of lactic and acetic acid, 10.04 g·L^-1 and 2.82 g·L^-1, respectively, were observed at 2 h of cultivation using glucose as the sole carbon source. Furthermore, the fermentation broth obtained using 1% β-glucan as the sole carbon source had greater antibacterial activity against selected pathogenic bacteria (B. subtilis TISTR 008, E. coli TISTR 780, and S. typhimurium TISTR 292) than did the broths prepared using glucose or FOS as the sole carbon source. The fermentation broth obtained by growing L. acidophilus BCC 13938 in the presence of β-glucan inhibited the growth of B. subtilis TISTR 008 by more than 70% and inhibited the growth of both S. typhimurium TISTR 292 and E. coli TISTR 780 by more than 90%. In conclusion, O. dipterigena BCC 2073 is a potential source of a β-glucan prebiotic that could be used for commercial production in the near future.

Keywords: β-glucan, Ophiocordyceps dipterigena, prebiotic, probiotic, antimicrobial.

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Authors: Tzong Yuan Wu, Sheng Kuo Hsieh, Tzyy Rong Jinn

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Fip-gts, an immunomodulatory protein purified from Ganoderma tsugae, has been reported to possess therapeutic effects in the treatment of cancer and autoimmune disease. For medicinal application, a recombinant Fip-gts was successfully expressed and purified in Sf21 insect cells by our previously work. It is important to evaluate the immunomodulatory activity of the rFip-gts. To assess the immunomodulatory potential of rFip-gts, the T lymphocytes of murine splenocytes were used in the present study. Results revealed that rFip-gts induced cellular aggregation formation. Additionally, the expression of IL-2 and IFN-r were up-regulated after the treatment of rFip-gts, and a corresponding increased production of IL-2 and IFN-r in a dose-dependent manner. The results showed that rFip-gts has an immunomodulatory activity in inducing Th1 lymphocytes from murine splenocytes released IL-2 and IFN-γ, thus suggest that rFip-gts may have therapeutic potential in vivo as an immune modulator.

Keywords: Fungal immunomodulatory protein, Ganodermatsugae, Interleukin 2, Interferon γ, Lingzhi.

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Authors: Klementina Puto, Ermelinda Nexhipi, Evi Llaka

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Dairy by-products are a fairly good environment for microorganisms due to their composition for their growth. Microbial populations have a significant impact in the production of cheese, butter, yogurt, etc. in terms of their organoleptic quality and at the same time some also cause their breakdown. In this paper, the microbiological contamination of soft cheese, butter and yogurt produced in the country (domestic) and imported is assessed, as an indicator of hygiene with impact on public health. The study was extended during September 2018-June 2019 and was divided into three periods, September-December, January-March, and April-June. During this study, a total of 120 samples were analyzed, of which 60 samples of cheese and butter locally produced, and 60 samples of imported soft cheese and butter productions. The microbial indicators analyzed are Staphylococcus aureus and E. coli. Analyzes have been conducted at the Food Safety Laboratory (FSIV) in Tirana in accordance with EU Regulation 2073/2005. Sampling was performed according to the specific international standards for these products (ISO 6887 and ISO 8261). Sampling and transport of samples were done under sterile conditions. Also, coding of samples was done to preserve the anonymity of subjects. After the analysis, the country's soft cheese products compared to imports were more contaminated with S. aureus and E. coli. Meanwhile, the imported butter samples that were analyzed, resulted within norms compared to domestic ones. Based on the results, it was concluded that the microbial quality of samples of cheese, butter and yogurt analyzed remains a real problem for hygiene in Albania. The study will also serve business operators in Albania to improve their work to ensure good hygiene on the basis of the HACCP plan and to provide a guarantee of consumer health.

Keywords: Consumer, health, dairy, by-products, microbial.

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Authors: Rupinder Bakshi, Geeta Walia, Anita Gupta

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Urinary Tract Infections are considered as one of the most common bacterial infections with an estimated annual global incidence of 150 million. Antimicrobial drug resistance is one of the major threats due to wide spread usage of uncontrolled antibiotics. In this study, a total number of 9149 urine samples were collected from R.H Patiala and processed in the Department of Microbiology G. M. C Patiala (January 2013 to December 2013). Urine samples were inoculated on MacConkey’s and blood agar plates and incubated at 370C for 24 hrs. The organisms were identified by colony characters, Gram’s staining, and biochemical reactions. Antimicrobial susceptibility of the isolates was determined against various antimicrobial agents (Hi – Media Mumbai India) by Kirby Bauer DISK diffusion method on Muller Hinton agar plates. Maximum patients were in the age group of 21-30 yrs followed by 31-40 yrs. Males (34%) are less prone to urinary tract infections than females (66%). Culture was positive in 25% of the samples. Escherichia coli was the most common isolate 60.3% followed by Klebsiella pneumoniae 13.5%, Proteus spp. 9% and Staphylococcus aureus 7.6%. Most of the urinary isolates were sensitive to, carbepenems, Aztreonam, Amikacin, and Piperacillin + Tazobactum. All the isolates showed a good sensitivity towards Nitrofurantoin (82%). ESBL production was found to be 70.6% in Escherichia coli and 29.4% in Klebsiella pneumonia. Susceptibility of ESBL producers to Imipenem, Nitrofurantoin and Amikacin were found to be 100%, 76%, and 75% respectively. Uropathogens are increasingly showing resistance to many antibiotics making empiric management of outpatient UTIs challenging. Ampicillin, Cotrimoxazole and Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin could be used in lower urinary tract infection. Knowledge of uropathogens and their antimicrobial susceptibility pattern in a geographical region will help in appropriate and judicious antibiotic usage in a health care setup.

Keywords: Urinary Tract Infection, UTI, antibiotic susceptibility pattern, ESBL.

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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: Douglas N. Simões, Michele Pittol, Vanda F. Ribeiro, Daiane Tomacheski, Ruth M. C. Santana

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The increasing demand of thermoplastic elastomers is related to the wide range of applications, such as automotive, footwear, wire and cable industries, adhesives and medical devices, cell phones, sporting goods, toys and others. These materials are susceptible to microbial attack. Moisture and organic matter present in some areas (such as shower area and sink), provide favorable conditions for microbial proliferation, which contributes to the spread of diseases and reduces the product life cycle. Compounds based on SEBS copolymers, poly(styrene-b-(ethylene-co-butylene)-b-styrene, are a class of thermoplastic elastomers (TPE), fully recyclable and largely used in domestic appliances like bath mats and tooth brushes (soft touch). Zinc oxide and zinc ions loaded in personal and home care products have become common in the last years due to its biocidal effect. In that sense, the aim of this study was to evaluate the effect of zinc as antimicrobial agent in compounds based on SEBS/polypropylene/oil/ calcite for use as refrigerator seals (gaskets), bath mats and sink squeegee. Two zinc oxides from different suppliers (ZnO-Pe and ZnO-WR) and one masterbatch of zinc ions (M-Zn-ion) were used in proportions of 0%, 1%, 3% and 5%. The compounds were prepared using a co-rotating double screw extruder (L/D ratio of 40/1 and 16 mm screw diameter). The extrusion parameters were kept constant for all materials. Tests specimens were prepared using the injection molding machine. A compound with no antimicrobial additive (standard) was also tested. Compounds were characterized by physical (density), mechanical (hardness and tensile properties) and rheological properties (melt flow rate - MFR). The Japan Industrial Standard (JIS) Z 2801:2010 was applied to evaluate antibacterial properties against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). The Brazilian Association of Technical Standards (ABNT) NBR 15275:2014 were used to evaluate antifungal properties against Aspergillus niger (A. niger), Aureobasidium pullulans (A. pullulans), Candida albicans (C. albicans), and Penicillium chrysogenum (P. chrysogenum). The microbiological assay showed a reduction over 42% in E. coli and over 49% in S. aureus population. The tests with fungi showed inconclusive results because the sample without zinc also demonstrated an inhibition of fungal development when tested against A. pullulans, C. albicans and P. chrysogenum. In addition, the zinc loaded samples showed worse results than the standard sample when tested against A. niger. The zinc addition did not show significant variation in mechanical properties. However, the density values increased with the rise in ZnO additives concentration, and had a little decrease in M-Zn-ion samples. Also, there were differences in the MFR results in all compounds compared to the standard.

Keywords: Antimicrobial, home device, SEBS, zinc.

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Authors: Moatlhodi Wise Letshwenyo, Lesedi Lebogang

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Botswana is an arid country that needs to start reusing wastewater as part of its water security plan. Pilot scale slow sand filtration in combination with roughing filter was investigated for the treatment of effluent from Botswana International University of Science and Technology to meet Botswana irrigation standards. The system was operated at hydraulic loading rates of 0.04 m/hr and 0.12 m/hr. The results show that the system was able to reduce turbidity from 262 Nephelometric Turbidity Units to a range between 18 and 0 Nephelometric Turbidity Units which was below 30 Nephelometric Turbidity Units threshold limit. The overall efficacy ranged between 61% and 100%. Suspended solids, Biochemical Oxygen Demand, and Chemical Oxygen Demand removal efficiency averaged 42.6%, 45.5%, and 77% respectively and all within irrigation standards. Other physio-chemical parameters were within irrigation standards except for bicarbonate ion which averaged 297.7±44 mg L^-1 in the influent and 196.22±50 mg L^-1 in the effluent which was above the limit of 92 mg L^-1, therefore averaging a reduction of 34.1% by the system. Total coliforms, fecal coliforms, and Escherichia coli in the effluent were initially averaging 1.1 log counts, 0.5 log counts, and 1.3 log counts respectively compared to corresponding influent log counts of 3.4, 2.7 and 4.1, respectively. As time passed, it was observed that only roughing filter was able to reach reductions of 97.5%, 86% and 100% respectively for faecal coliforms, Escherichia coli, and total coliforms. These organism numbers were observed to have increased in slow sand filter effluent suggesting multiplication in the tank. Water quality index value of 22.79 for the physio-chemical parameters suggests that the effluent is of excellent quality and can be used for irrigation purposes. However, the water quality index value for the microbial parameters (1820) renders the quality unsuitable for irrigation. It is concluded that slow sand filtration in combination with roughing filter is a viable option for the treatment of secondary effluent for reuse purposes. However, further studies should be conducted especially for the removal of microbial parameters using the system.

Keywords: Irrigation, roughing filter, slow sand filter, turbidity, water quality index.

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Authors: Eugenie A. Kachvoryan, Astghik Z. Pepoyan, Maria V. Harutyunova, Anahit M. Manvelyan

Abstract:

Taking into account the importance of Lake Sevan and Lake Sevan basin-s rivers for Armenian economy, the main goals of our investigations were the documentation of water quality and the biodiversity of invertebrates developed in Lake Sevan basin-s rivers and selected tributaries. Moderately satisfied ecological condition for the biodiversity of Lake Sevan basin-s rivers has been established, and the changes in species- composition of zoobenthos in Lake Sevan were detected. A growing tendency of antibiotic resistance among E. coli isolates in water resources has been shown.

Keywords: Biodiversity, ecosystem, Lake Sevan, water-quality, zoobenthos.

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Authors: Seung Pil Pack

Abstract:

Carbonic anhydrases (CAs) has been focused as biological catalysis for CO2 sequestration process because it can catalyze the conversion of CO2 to bicarbonate. Here, codon-optimized sequence of α type-CA cloned from Duneliala species. (DsCAopt) was constructed, expressed, and characterized. The expression level in E. coli BL21(DE3) was better for codon-optimized DsCAopt than intact sequence of DsCAopt. DsCAopt enzyme shows high-stability at pH 7.6/10.0. In final, we demonstrated that in the Ca2+ solution, DsCAopt enzyme can catalyze well the conversion of CO2 to CaCO3, as the calcite form.

Keywords: Carbonic anhydrase, Codon-optimization, Duneliala species, CO2 sequestration

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Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

Abstract:

One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures.

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Authors: Abdulwahab Kammon, Tan Sheau Wei, Abdul Rahman Omar, Abdunaser Dayhum, Ibrahim Eldghayes, Monier Sharif

Abstract:

Infectious bronchitis virus (IBV) is a very dynamic and evolving virus, causing major economic losses to the global poultry industry. Recently, the Libyan poultry industry faced severe outbreak of respiratory distress associated with high mortality and dramatic drop in egg production. Tracheal and cloacal swabs were analyzed for several poultry viruses. IBV was detected using SYBR Green I real-time PCR detection based on the nucleocapsid (N) gene. Sequence analysis of the partial N gene indicated high similarity (~ 94%) to IBV strain 3382/06 that was isolated from Taiwan. Even though the IBV strain 3382/06 is more similar to that of the Mass type H120, the isolate has been implicated associated with intertypic recombinant of 3 putative parental IBV strains namely H120, Taiwan strain 1171/92 and China strain CK/CH/LDL/97I. Complete sequencing and antigenicity studies of the Libya IBV strains are currently underway to determine the evolution of the virus and its importance in vaccine induced immunity. In this paper we documented for the first time the presence of possibly variant IBV strain from Libya which required dramatic change in vaccination program.

Keywords: Libya, Infectious bronchitis, Molecular characterization.

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Authors: Tahereh Naji, Mahsa Jassemi

Abstract:

In this investigation, the antibacterial effects of ethanolic and 7:3 isopropyl –hexane mixture extracts of Zingiber officinale were evaluated against three Gram positive bacteria, B. cereus, S.epidermidis, S. aureus and three Gram negative bacteria, E. coli, K.pneumonia and P.areuginosa. Utilizing paper disk diffusion and well methods in-vitro, MIC and MBC were determined by macrodilution. The results showed that ethanolic rhizome extract of ginger had significantly active than Isopropyl –hexan extract. Further work needs to be done in these extracts including fractionation to isolate active constituents and subsequent pharmacological evaluation.

Keywords: Antibacterial, Medicinal plant extract, Zingiberofficinale.

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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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Authors: Hai Xu, Yiwei Wang, Xi Bao, Bihua Deng, Pengcheng Li, Yu Lu

Abstract:

T7 phage could be used as a perfect vector for peptides expression and haptens presentation. T7-3GnRH recombinant phage was constructed by inserting three copies of Gonadotrophin Releasing Hormone (GnRH) gene into the multiple cloning site of T7 Select 415-1b phage genome. The positive T7-3GnRH phage was selected by using polymerase chain reaction amplification, and the p10B-3GnRH fusion protein was verified by SDS-PAGE and Western-blotting assay. T7-3GnRH vaccine was made and immunized with 10¹⁰ pfu in 0.2 ml per dose in mice. Blood samples were collected at an interval in weeks, and anti-GnRH antibody and testosterone concentrations were detected by ELISA and radioimmunoassay, respectively. The results show that T7-3GnRH phage particles confer a high immunogenicity to the GnRH-derived epitope. Moreover, the T7-3GnRH vaccine induced higher level of anti-GnRH antibody than ImproVac^®. However, the testosterone concentrations in both immunized groups were at a similar level, and the testis developments were significantly inhibited compared to controls. These findings demonstrated that the anti-GnRH antibody could neutralize the endogenous GnRH to down regulate testosterone level and limit testis development, highlighting the potential value of T7-3GnRH in the immunocastration vaccine research.

Keywords: Gonadotrophin releasing hormone, GnRH, immunocastration, T7 phage, phage vaccine.

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Authors: Mona Heydari, Ehsan Motamedian, Seyed Abbas Shojaosadati

Abstract:

Prediction of perturbations after genetic manipulation (especially gene knockout) is one of the important challenges in systems biology. In this paper, a new algorithm is introduced that integrates microarray data into the metabolic model. The algorithm was used to study the change in the cell phenotype after knockout of Gss gene in Escherichia coli BW25113. Algorithm implementation indicated that gene deletion resulted in more activation of the metabolic network. Growth yield was more and less regulating gene were identified for mutant in comparison with the wild-type strain.

Keywords: Metabolic network, gene knockout, flux balance analysis, microarray data, integration.

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Authors: Hiba Hasan, Khalid Raza

Abstract:

Reverse engineering of genetic regulatory network involves the modeling of the given gene expression data into a form of the network. Computationally it is possible to have the relationships between genes, so called gene regulatory networks (GRNs), that can help to find the genomics and proteomics based diagnostic approach for any disease. In this paper, clustering based method has been used to reconstruct genetic regulatory network from time series gene expression data. Supercoiled data set from Escherichia coli has been taken to demonstrate the proposed method.

Keywords: Gene expression, gene regulatory networks (GRNs), clustering, data preprocessing, network visualization.

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