Search results for: bacterial susceptibility
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1516

Search results for: bacterial susceptibility

796 Focus on the Bactericidal Efficacies of Alkaline Agents in Solid and the Required Time for Bacterial Inactivation

Authors: Hakimullah Hakim, Chiharu Toyofuku, Mari Ota, Mayuko Suzuki, Miyuki Komura, Masashi Yamada, Md. Shahin Alam, Natthanan Sangsriratanakul, Dany Shoham, Kazuaki Takehara

Abstract:

Disinfectants and their application are essential part of infection control strategies and enhancement of biosecurity at farms, worldwide. Alkaline agents are well known for their strong and long term antimicrobial capacities and most frequently are applied at farms for control and prevention of biological hazards. However, inadequate information regarding such materials’ capacities to inactivate pathogens and their improper applications fail farmers to achieve the mentioned goal. Thus, this requires attention to further evaluate their efficacies, under different conditions and in different ways. Here in this study we evaluated bactericidal efficacies of food additive grade of calcium hydroxide (FdCa(OH)2) powder derived from natural calcium carbonates obtained from limestone (Fine Co., Ltd., Tokyo, Japan), and bioceramic powder (BCX) derived from chicken feces at pH 13 (NMG environmental development Co., Ltd., Tokyo, Japan), for their efficacies to inactivate bacteria in feces. [Materials & Methods] Chicken feces were inoculated by 100 µl Escherichia coli and Salmonella Infantis in falcon tubes, individually, then FdCa(OH)2 or BCX powders were individually added to make final concentration of 0, 5, 10, 20 and 30% (w/w) in total weight of 0.5g, followed by properly mixing and incubating at room temperature for certain periods of time, in a dark place. Afterwards, 10 ml 1M Tris-HCl (pH 7.2) was added onto them to reduce their pH, in order to stop powders’ activities and to harvest the remained viable bacteria, whereas using normal medium or dW2 to recover bacteria increases the mixture pH, and as a result bacteria would be inactivated soon; therefore, the latter practice brings about incorrect and misleading results. Samples were then inoculated on DHL agar plates in order to calculate colony forming units (CFU)/ml of viable bacteria. [Results and Discussion] FdCa(OH)2 powder at 10% and 5% required 3 hr and 6 hr exposure times, respectively, while BCX powder at 20% concentrations required 6 hr exposure time to kill the mentioned bacteria in feces down to lower than detectable level (≤ 3.6 log10 CFU/ml). This study confirmed capacities of FdCa(OH)2 and BCX powders to inactivate bacteria in feces, and both materials are environment friendly materials, with no risk to human or animal’s health. This finding helps farmers to properly apply alkaline agents in appropriate concentrations and exposure times in their farms, in order to prevent and control infectious diseases outbreaks and to enhance biosecurity. Finally, this finding may help farmers to implement better strategies for infections control in their livestock farms.

Keywords: bacterial inactivation, bioceramic, biosecurity at livestock farms, chicken feces

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

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

Abstract:

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

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

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794 Infestations of Olive Fruit Fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), in Different Olive Cultivars in Çanakkale, Turkey

Authors: Hanife Genç

Abstract:

The olive fruit fly, Bactrocera oleae (Rossi), is an economically important and endemic pest in olive (Oleae europae) orchards in Turkey. The aim of this study was to determine olive fruit fly infestation in different olive cultivars in the laboratory. Olive fly infested fruits were collected in Çanakkale province to establish wild fly population. After having reproductive olive fly colonies, 14 olive cultivars were tested in the controlled laboratory conditions, at 23±2 °C, 65% RH and 16:8 h (light: dark) photoperiod. The olive samples from 14 different olive cultivars were collected in October 2015, in Campus of Dardanos, Çanakkale Onsekiz Mart University. Observations were carried out detecting some biological parameters such as the number of oviposition stings, active infestation, total infestation, the number of pupae and the adult emergence. The results indicated that oviposition stings were not associated with pupal yield. A few pupae were found within olive fruits which were not able to exit. Screening of the varieties suggested that less susceptible cultivar to olive fruit fly attacks was Arbequin while Gemlik-2M 2/3 showed significant susceptibility. Ovipositional preference of olive fly females and the success of larval development in different olive varieties are crucial for establishing new olive orchards to prevent high olive fruit fly infestation.

Keywords: infestation, olive fruit fly, olive cultivars, oviposition sting

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793 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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792 Evaluation of Ecological Resilience in Mountain-plain Transition Zones: A Case Study of Dujiangyan City, Chengdu

Authors: Zhu Zhizheng, Huang Yong, Li Tong

Abstract:

In the context of land and space development and resource environmental protection. Due to its special geographical location, mountain-plain transition zones are limited by many factors such as topography, mountain forest protection, etc., and their ecology is also more sensitive, with the characteristics of disaster susceptibility and resource gradient. Taking Dujiangyan City, Chengdu as an example, this paper establishes resilience evaluation indicators on the basis of ecological suitability evaluation through the analysis of current situation data and relevant policies: water conservation evaluation, soil and water conservation evaluation, biodiversity evaluation, soil erosion sensitivity evaluation, etc. Based on GIS spatial analysis, the ecological suitability and resilience evaluation results of Dujiangyan city were obtained by disjunction operation. The ecological resilience level of Dujiangyan city was divided into three categories: high, medium and low, with an area ratio of 50.81%, 16.4% and 32.79%, respectively. This paper can provide ideas for solving the contradiction between man and land in the mountain-plain transition zones, and also provide a certain basis for the construction of regional ecological protection and the delineation of three zones and three lines.

Keywords: urban and rural planning, ecological resilience, dujiangyan city, mountain-plain transition zones

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791 In Silico Study of the Biological and Pharmacological Activity of Nigella sativa

Authors: Ammar Ouahab, Meriem Houichi , Sanna Mihoubi

Abstract:

Background: Nigella sativa is an annual flowering plant, belongs to the Ranunculaceae family. It has many pharmacological activities such as anti-inflammatory; anti-bacterial; anti-hepatotoxic activities etc. Materials: In order to predict the pharmacological activity of Nigella Sativa’s compounds, some web based servers were used, namely, PubChem, Molinspiration, ADMET-SAR, PASS online and PharMapper. In addition to that, AutoDOCK was used to investigate the different molecular interactions between the selected compounds and their target proteins. Results: All compounds displayed a stable interaction with their targets and satisfactory binding energies, which means that they are active on their targets. Conclusion: Nigella sativa is an effective medicinal plant that has several ethno-medical uses; the latter uses are proven herein via an in-silico study of their pharmacological activities.

Keywords: Nigella sativa, AutoDOCK, PubChem, Molinspiration, ADMET-SAR, PharMapper, PASS online server, docking

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790 Prevalence and Potential Risk Factors Associated with Skin Affection in Donkeys

Authors: Mohamed Z. Sayed-Ahmed, Ahmed M. Ahdy, Emad E. Younis, Sabry A. El-Khodary

Abstract:

Little research information is available on the prevalence of diseases of donkeys in Egypt. Across sectional study was undertaken between March 2009 and February 2010 to verify the prevalence of skin affection of donkeys. A total of 1134 donkeys in northern Egypt were investigated. A questionnaire was constructed to verify the number of infected contact animals as well as the associated factors. Physical examination was carried out, and the distribution of skin lesions was recorded. Skin scraping and biopsy were obtained to perform bacteriological, mycological, and histopathological examinations. Thirty-five (3.09%) out of 1134 noticed donkeys had skin affections including mange (18/35), dermatophytosis (6/35), bacterial dermatitis (6/35) urticaria (2/35) and allergic dermatitis (3/35). The present results indicate that mange and dermatophytosis are the prevalent skin diseases in donkeys. Contact with other animal species of contaminated environment may contribute to the occurrence of the diseases.

Keywords: donkeys, Egypt, prevalence, skin affection

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789 Development of Gold Nanoparticles-Antibody System for the Selective Photothermal Destruction of Multidrug Resistant Bacteria

Authors: Teodora Mocan, Lucian Mocan, Cornel Iancu, Flaviu A. Tabaran, Bartos Dana, Matea Cristian

Abstract:

Antimicrobial resistance, which threatens the efficacy of the existing antibiotics represents a worldwide public health issue. At the current time, vancomycin is the only responsive treatment although has significant cytotoxicity, is partially effective and it is poorly retained by infected tissues. From a clinical point of view, attractive alternative approaches for treating such Meticillin-Resistant Staphylococcus Aureus (MRSA) strains would be using agents that cause physical damage to the bacteria. Modular nanopharmaceuticals systems are being designed to address all of these multifunctional capabilities for the ideal bacterial treatment, with the ability to mix and match appropriate functions. Here we present a novel method of selective laser photothermal ablation of MRSA bacteria mediated by gold nanoparticles bound to PBP antibody against PBP protein located on the MRSA surface.

Keywords: MRSA, laser, nanoparticle, antibody

Procedia PDF Downloads 259
788 Molecular Identification of Camel Tick and Investigation of Its Natural Infection by Rickettsia and Borrelia in Saudi Arabia

Authors: Reem Alajmi, Hind Al Harbi, Tahany Ayaad, Zainab Al Musawi

Abstract:

Hard ticks Hyalomma spp. (family: Ixodidae) are obligate ectoparasite in their all life stages on some domestic animals mainly camels and cattle. Ticks may lead to many economic and public health problems because of their blood feeding behavior. Also, they act as vectors for many bacterial, viral and protozoan agents which may cause serious diseases such as tick-born encephalitis, Rocky-mountain spotted fever, Q-fever and Lyme disease which can affect human and/or animals. In the present study, molecular identification of ticks that attack camels in Riyadh region, Saudi Arabia based on the partial sequence of mitochondrial 16s rRNA gene was applied. Also, the present study aims to detect natural infections of collected camel ticks with Rickessia spp. and Borelia spp. using PCR/hybridization of Citrate synthase encoding gene present in bacterial cells. Hard ticks infesting camels were collected from different camels located in a farm in Riyadh region, Saudi Arabia. Results of the present study showed that the collected specimens belong to two species: Hyalomma dromedari represent 99% of the identified specimens and Hyalomma marginatum which account for 1 % of identified ticks. The molecular identification was made through blasting the obtained sequence of this study with sequences already present and identified in GeneBank. All obtained sequences of H. dromedarii specimens showed 97-100% identity with the same gene sequence of the same species (Accession # L34306.1) which was used as a reference. Meanwhile, no intraspecific variations of H. marginatum mesured because only one specimen was collected. Results also had shown that the intraspecific variability between individuals of H. dromedarii obtained in 92 % of samples ranging from 0.2- 6.6%, while the remaining 7 % of the total samples of H. dromedarii showed about 10.3 % individual differences. However, the interspecific variability between H. dromedarii and H. marginatum was approximately 18.3 %. On the other hand, by using the technique of PCR/hybridization, we could detect natural infection of camel ticks with Rickettsia spp. and Borrelia spp. Results revealed the natural presence of both bacteria in collected ticks. Rickettsial spp. infection present in 29% of collected ticks, while 35% of collected specimen were infected with Borrelia spp. The valuable results obtained from the present study are a new record for the molecular identification of camel ticks in Riyadh, Saudi Arabia and their natural infection with both Rickettsia spp. and Borrelia spp. These results may help scientists to provide a good and direct control strategy of ticks in order to protect one of the most important economic animals which are camels. Also results of this project spotlight on the disease that might be transmitted by ticks to put out a direct protective plan to prevent spreading of these dangerous agents. Further molecular studies are needed to confirm the results of the present study by using other mitochondrial and nuclear genes for tick identification.

Keywords: Camel ticks, Rickessia spp. , Borelia spp. , mitochondrial 16s rRNA gene

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787 Carrot: A Possible Source of Multidrug-Resistant Acinetobacter Transmission

Authors: M. Dahiru, O. I. Enabulele

Abstract:

The research wish to investigate the occurrence of multidrug- resistant Acinetobacter, in carrot and estimate the role of carrot in its transmission, in a rapidly growing urban population. Thus, 50 carrot samples were collected from Jakara wastewater irrigation farms and analyzed on MacConkey agar and screened by Microbact 24E (Oxoid) and susceptibility of isolates tested against 10 commonly used antibiotics. Acinetobacter baumannii and A. lwoffii were isolated in 22.00% and 16% of samples respectively. Resistance to ceporex and penicillin of 36.36% and 27.27% in A. baumannii, and sensitivity to ofloxacin, pefloxacin, gentimycin and co-trimoxazole, were observed. However, for A. lwoffii apart from 37.50% resistance to ceporex, it was also resistant to all other drugs tested. There was a similarity in the resistant shown by A. baumannii and A. lwoffii to fluoroquinolones drugs and β- lactame drugs families in addition to between sulfonamide and animoglycoside demonstrated by A. lwoffii. Interestingly, when resistant similarities to different antibiotics were compared for A. baumannii and A. lwoffii as a whole, significant correlation was observed at P < 0.05 to CPX to NA (46.2%), and SXT to AU (52.6%) respectively, and high multi drug resistance (MDR) of 27.27% and 62.50% by A. baumannii and A. lwoffii respectively and overall MDR of 42.11% in all isolates. The occurrence of multidrug-resistance pathogen in carrot is a serious challenge to public health care, especially in a rapidly growing urban population where subsistence agriculture contributes greatly to urban livelihood and source of vegetables.

Keywords: urban agriculture, public health, fluoroquinolone, sulfonamide, multidrug-resistance

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786 Influence of S.carnosus Bacteria as Biocollector for the Recovery Organic Matter in the Flotation Process

Authors: G. T. Ramos-Escobedo, E. T. Pecina-Treviño, L. F. Camacho-Ortegon, E. Orrantia-Borunda

Abstract:

The mineral bioflotation represents a viable alternative for the evaluation of new processes benefit alternative. The adsorption bacteria on minerals surfaces will depend mainly on the type of the microorganism as well as of the studied mineral surface. In the current study, adhesion of S. carnosus on coal was studied. Several methods were used as: DRX, Fourier Transform Infra Red (FTIR) adhesion isotherms and kinetic. The main goal is the recovery of organic matter by the microflotation process on coal particles with biological reagent (S. carnosus). Adhesion tests revealed that adhesion took place after 8 h at pH 9. The results suggest that the adhesion of bacteria to solid substrates can be considered an abiotic physicochemical process that is consequently governed by bacterial surface properties such as their specific surface area, hydrophobicity and surface functionalities. The greatest coal fine flotability was 75%, after 5 min of flotation.

Keywords: fine coal, bacteria, adhesion, recovery organic matter

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785 Differential Infection of Primary Human B-Cells and EBV Positive B-Lymphoma Cell Lines by Recombinant AAV Serotypes

Authors: Elham Ahmadi, Mehrdad Ravanshad, Joyce Fingeroth, Mazyar Ziyaeyan, Rajesh Panigrahi, Jun Xie, Gao Guangping

Abstract:

B-cell proliferative disorders often occur among persons that are T-cell compromised. These disorders are primarily EBV+ and can first present with a focal lesion. Direct introduction of oncolytic viruses into localized tumors provides theoretical advantages over chemotherapy and immunotherapy by reducing systemic toxicity, to which the immunocompromised host is most vulnerable. Widely studied as a vehicle for gene therapy, AAV has only rarely been applied to treat cancer. As a prelude to development of a therapeutic vehicle, we assessed the ability of 15 distinct recombinant AAV serotypes (rAAV1, rAAV2, rAAV3b, rAAV4, rAAV5, rAAV6, rAAV6.2, rAAV6TM, rAAV7, rAAV8, rAAVrh8, rAAV9, rAAVrh10, rAAV39, rAAV43) bearing eGFP to infect human B-cell tumor lines compared with primary B-cells in vitro. Enhanced infection of tumor lines by AAV 6.2 was demonstrated by flow cytometry. EBV superinfection of EBV negative B-cell tumor lines increased susceptibility to AAV6.2 infection. As proof of concept, AAV6.2 bearing HSV-1 thymidine kinase in place of eGFP eliminated tumor cells upon exposure to ganciclovir.

Keywords: AAV, gene therapy, lymphoma, malignancy, tropism

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784 Virtualization of Biomass Colonization: Potential of Application in Precision Medicine

Authors: Maria Valeria De Bonis, Gianpaolo Ruocco

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Nowadays, computational modeling is paving new design and verification ways in a number of industrial sectors. The technology is ripe to challenge some case in the Bioengineering and Medicine frameworks: for example, looking at the strategical and ethical importance of oncology research, efforts should be made to yield new and powerful resources to tumor knowledge and understanding. With these driving motivations, we approach this gigantic problem by using some standard engineering tools such as the mathematics behind the biomass transfer. We present here some bacterial colonization studies in complex structures. As strong analogies hold with some tumor proliferation, we extend our study to a benchmark case of solid tumor. By means of a commercial software, we model biomass and energy evolution in arbitrary media. The approach will be useful to cast virtualization cases of cancer growth in human organs, while augmented reality tools will be used to yield for a realistic aid to informed decision in treatment and surgery.

Keywords: bacteria, simulation, tumor, precision medicine

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783 Thermomagnetic Convection of a Ferrofluid in a Non-Uniform Magnetic Field Induced a Current Carrying Wire

Authors: Ashkan Vatani, Peter Woodfield, Nam-Trung Nguyen, Dzung Dao

Abstract:

Thermomagnetic convection of a ferrofluid flow induced by the non-uniform magnetic field around a current-carrying wire was theoretically analyzed and experimentally tested. To show this phenomenon, the temperature rise of a hot wire, immersed in DIW and Ferrofluid, as a result of joule heating has been measured using a transient hot-wire technique. When current is applied to the wire, a temperature gradient is imposed on the magnetic fluid resulting in non-uniform magnetic susceptibility of the ferrofluid that results in a non-uniform magnetic body force which makes the ferrofluid flow as a bulk suspension. For the case of the wire immersed in DIW, free convection is the only means of cooling, while for the case of ferrofluid a combination of both free convection and thermomagnetic convection is expected to enhance the heat transfer from the wire beyond that of DIW. Experimental results at different temperatures and for a range of constant currents applied to the wire show that thermomagnetic convection becomes effective for the currents higher than 1.5A at all temperatures. It is observed that the onset of thermomagnetic convection is directly proportional to the current applied to the wire and that the thermomagnetic convection happens much faster than the free convection. Calculations show that a 35% enhancement in heat transfer can be expected for the ferrofluid compared to DIW, for a 3A current applied to the wire.

Keywords: cooling, ferrofluid, thermomagnetic convection, magnetic field

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782 Development and Evaluation of Antimicrobial Herbal Mouthwash Including Methanolic Extracts of Beautea monosperma and Cordia obliqua

Authors: Reenu Yadav, S. K. Yadav

Abstract:

Herbal therapy has been used for daily oral health care to prevent, treat or cure oral conditions from halitosis to periodontal diseases. The importance of mouth and teeth cleanliness has been recognized from the earliest days of civilization to the 21st century. In the present study, leaves and seeds of Cordia obliqua and barks and twigs of Beautea monosperma, which is used traditionally for oral diseases was evaluated for its antimicrobial activity. The antimicrobial activity tests indicated that the methanolic extract exhibited stronger activities against the commonly encountered oral bacterial and fungal pathogens. The mouthwash formulation prepared and it is compared with marketed formulation HiOra. The results indicated that the herbal mouthwash could inhibit the growth of oral pathogens and may prevent plaque and other periodontal diseases caused by dental pathogens.

Keywords: herbal mouthwash, bio medicine, life sciences, herbal extracts

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781 Phytochemical Screening and Antibacterial Activities of Tapinanthus dodoneifolius Leaves Extracts against Some Selected Clinical Isolates

Authors: Isa Usman Balan, Umar Aliyu, Ahmad Tijjani Muhammed

Abstract:

The laboratory scale experiment was conducted to determine the phytochemical constituents and antibacterial activities of epiphytic neem leaves (Tapinanthusdodoneifolius) extracts on some selected clinical isolates. The samples were collected using polythene bags to avoid unnecessary contamination of the plants, and they were collected from the old site garden of the BUK. The phytochemical screening and antibacterial test were carried out in the Chemistry and Biology laboratory, respectively at Bayero University Kano (BUK). The result obtained showed that carbohydrates, glycosides, steroids, alkaloids, phenol, saponins and flavonoids are present in the ethanolic extract. However, chloroform extract showed only glycosides, phenols, and carbohydrates. Furthermore, there was no significant difference between the ethanolic extracts and bacterial isolates (p<0.05).

Keywords: phytochemical screening, antibacterial, clinical isolates, epiphytic neem leaves, Tapinanthus dodoneifolius

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780 Influence of Cation Substitution on Magnetic Transitions and Ordering in La2NixCo1-xMnO6 Compounds (x = 0.2 - 0.8)

Authors: Amine.Harbia, Hicham. Moutaabbidb, Yann. Le Godecb, Said. Benmokhtara, Mouhammed. Moutaabbida

Abstract:

This study explores the structural and magnetic characteristics of newly synthesized double perovskite oxides, La₂NiₓCo1-xMnO₆, with x ranging from 0.2 to 0.8. Utilizing X-ray powder diffraction and SQUID magnetometry, we analyzed the compounds that consistently exhibit a monoclinic structure with the P21/n space group at ambient temperature. it findings reveal that as Ni2+ is progressively substituted by Co2+, there is a corresponding decrease in cell parameters, attributable to the smaller ionic radius of Ni2+ (0.69 Å) compared to Co2+ (0.74 Å). The crystal structure features octahedrally coordinated (Co/Ni)2+ and Mn4+ cations with oxygen, forming (Co/Ni)O6 and MnO6 octahedra linked via oxygen atoms along different crystallographic axes. Magnetic characterization conducted over a temperature range of 2 to 300 K in both DC and AC magnetic fields, showed a predominant paramagnetic to ferromagnetic transition between 232 K and 260 K, with the Curie temperature notably increasing with higher x values. Samples with x=0.2, 0.25, and 0.5 exhibited a secondary PM-FM transition between 200 K and 208 K. Cation ordering was quantitatively assessed, indicating a higher ordering in Ni2+-rich samples (x=0.75 and 0.8) at over 96%, whereas the sample with x=0.25 showed minimal ordering. Furthermore, the out-of-phase component of the AC susceptibility displayed frequency-dependent transitions between 65 K and 110 K, suggesting the presence of superparamagnetic domains across all samples.

Keywords: double perovskite oxides, magnetic transitions, cation ordering, squid magnetometry

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779 Characterization of Surface Microstructures on Bio-Based PLA Fabricated with Nano-Imprint Lithography

Authors: D. Bikiaris, M. Nerantzaki, I. Koliakou, A. Francone, N. Kehagias

Abstract:

In the present study, the formation of structures in poly(lactic acid) (PLA) has been investigated with respect to producing areas of regular, superficial features with dimensions comparable to those of cells or biological macromolecules. Nanoimprint lithography, a method of pattern replication in polymers, has been used for the production of features ranging from tens of micrometers, covering areas up to 1 cm², down to hundreds of nanometers. Both micro- and nano-structures were faithfully replicated. Potentially, PLA has wide uses within biomedical fields, from implantable medical devices, including screws and pins, to membrane applications, such as wound covers, and even as an injectable polymer for, for example, lipoatrophy. The possibility of fabricating structured PLA surfaces, with structures of the dimensions associated with cells or biological macro- molecules, is of interest in fields such as cellular engineering. Imprint-based technologies have demonstrated the ability to selectively imprint polymer films over large areas resulting in 3D imprints over flat, curved or pre-patterned surfaces. Here, we compare nano-patterned with nano-patterned by nanoimprint lithography (NIL) PLA film. A silicon nanostructured stamp (provided by Nanotypos company) having positive and negative protrusions was used to pattern PLA films by means of thermal NIL. The polymer film was heated from 40°C to 60°C above its Tg and embossed with a pressure of 60 bars for 3 min. The stamp and substrate were demolded at room temperature. Scanning electron microscope (SEM) images showed good replication fidelity of the replicated Si stamp. Contact-angle measurements suggested that positive microstructuring of the polymer (where features protrude from the polymer surface) produced a more hydrophilic surface than negative micro-structuring. The ability to structure the surface of the poly(lactic acid), allied to the polymer’s post-processing transparency and proven biocompatibility. Films produced in this were also shown to enhance the aligned attachment behavior and proliferation of Wharton’s Jelly Mesenchymal Stem cells, leading to the observed growth contact guidance. The bacterial attachment patterns of some bacteria, highlighted that the nano-patterned PLA structure can reduce the propensity for the bacteria to attach to the surface, with a greater bactericidal being demonstrated activity against the Staphylococcus aureus cells. These biocompatible, micro- and nanopatterned PLA surfaces could be useful for polymer– cell interaction experiments at dimensions at, or below, that of individual cells. Indeed, post-fabrication modification of the microstructured PLA surface, with materials such as collagen (which can further reduce the hydrophobicity of the surface), will extend the range of applications, possibly through the use of PLA’s inherent biodegradability. Further study is being undertaken to examine whether these structures promote cell growth on the polymer surface.

Keywords: poly(lactic acid), nano-imprint lithography, anti-bacterial properties, PLA

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778 Microbiological and Physicochemical Evaluation of Traditional Greek Kopanisti Cheese Produced by Different Starter Cultures

Authors: M. Kazou, A. Gavriil, O. Kalagkatsi, T. Paschos, E. Tsakalidou

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Kopanisti cheese is a Greek soft Protected Designation of Origin (PDO) cheese made of raw cow, sheep or goat milk, or mixtures of them, with similar organoleptic characteristics to that of Roquefort cheese. Traditional manufacturing of Kopanisti cheese is limited in small-scale dairies, without the addition of starter cultures. Instead, an amount of over-mature Kopanisti cheese, called Mana Kopanisti, is used to initiate ripening. Therefore, the selection of proper starter cultures and the understanding of the contribution of various microbial groups to its overall quality is crucial for the production of a high-quality final product with standardized organoleptic and physicochemical characteristics. Taking the above into account, the aim of the present study was the investigation of Kopanisti cheese microbiota and its role in cheese quality. For this purpose, four different types of Kopanisti were produced in triplicates, all with pasteurized cow milk, with the addition of (A) the typical mesophilic species Lactococcus lactis and Lactobacillus paracasei used as starters in the production of soft spread cheeses, (B) strains of Lactobacillus acidipiscis and Lactobacillus rennini previously isolated from Kopanisti and Mana Kopanisti, (C) all the species from (A) and (B) as inoculum, and finally (D) the species from (A) and Mana Kopanisti. Physicochemical and microbiological analysis was performed for milk and cheese samples during ripening. Enumeration was performed for major groups of lactic acid bacteria (LAB), total mesophilic bacteria, yeasts as well as hygiene indicator microorganisms. Bacterial isolates from all the different LAB groups, apart from enterococci, alongside yeasts isolates, were initially grouped using repetitive sequence-based polymerase chain reaction (rep-PCR) and then identified at the species level using 16S rRNA gene and internal transcribed spacer (ITS) DNA region sequencing, respectively. Sensory evaluation was also performed for final cheese samples at the end of the ripening period (35 days). Based on the results of the classical microbiological analysis, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, ranged between 7 and 10 log colony forming unit (CFU) g⁻¹, phychrotrophic bacteria, and yeast extract glucose chloramphenicol (YGC) isolates between 4 and 8 log CFU g⁻¹, while coliforms and enterococci up to 2 log CFU g⁻¹ throughout ripening in cheese samples A, C and D. In contrast, in cheese sample B, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, phychrotrophic bacteria, and YGC isolates ranged between 0 and 10 log CFU g⁻¹ and coliforms and enterococci up to 2 log CFU g⁻¹. Although the microbial counts were not that different among samples, identification of the bacterial and yeasts isolates revealed the complex microbial community structure present in each cheese sample. Differences in the physicochemical characteristics among the cheese samples were also observed, with pH ranging from 4.3 to 5.3 and moisture from 49.6 to 58.0 % in the final cheese products. Interestingly, the sensory evaluation also revealed differences among samples, with cheese sample B ranking first based on the total score. Overall, the combination of these analyses highlighted the impact of different starter cultures on the Kopanisti microbiota as well as on the physicochemical and sensory characteristics of the final product.

Keywords: Kopanisti cheese, microbiota, classical microbiological analysis, physicochemical analysis

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777 In vitro Antioxidant and Antibacterial Activities of Methanol Extracts of Tamus communis L. from Algeria

Authors: F. Belkhiri, A. Baghiani, S. Boumerfeg, N. Charef, S. Khennouf, L. Arrar

Abstract:

The present study was conducted to evaluate the in vitro antioxidant and antibacterial properties of methanolic extracts from roots of Tamus communis L. (TCRE), which is a plant used in traditional medicine in Algeria. The antioxidant potential of pattern was evaluated using tow complementary techniques, inhibition of free radical DPPH and the test of β-Carotene/linoleic acid. The antioxidant test indicates that non-polar fractions of TCRE (chloroform and ethyl acetate fractions) were more active than the polar fractions. Among these fractions, the chloroform extract appear in the DPPH test an IC50 of (18.89 µg/ml) comparable to that of BHT (18.6 µg/ml). This fraction was able to inhibiting the oxidation of β-Carotene with a percentage of inhibition (89.84 %). In antibacterial test, non-polar fractions showed antibacterial activity very important compared with the polar fractions. These fractions have inhibited the growth of four from nine bacterial strains, causing zones of inhibition from 08 to 23 mm of diameter.

Keywords: antioxidant activity, antibacterial activity, Tamus communis L., polar fractions

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776 An Invertebrate-Type Lysozyme from Chinese Mitten Crab Eriocheir Sinensis: Cloning and Characterization

Authors: Fengmei Li, Li Xu, Guoliang Xia

Abstract:

Lysozyme is a catalytic enzyme that performs bacterial cell lysis by cleaving the β-1,4-glycosidic bond between N-acetylmuramic acid and N-acetylglucosamine of peptidoglycan in cell walls. In the present study, an invertebrate-type (i-type) lysozyme gene was cloned from Chinese mitten crab Eriocheir sinensis (designated as EsLysozyme) based on PCR-based rapid amplification of cDNA ends (RACE) technology. The full-length cDNA of EsLysozyme was of 831 bp. SMART and SIGNALP 3.0 program analysis revealed that EsLysozyme contained a signal peptide and a destabilase domain. The five amino acid residues (Tyr63, Trp64, Tyr91, His110, Pro114) and the conserved motif GSLSCG(P/Y)FQI and CL(E/L/R/H)C(I/M)C in i-type lysozymes were also found in EsLysozyme. The high similarity of EsLysozyme with L. vannamei lysozymes and phylogenetic analysis suggested that EsLysozyme should be a new member of i-type lysozyme family.

Keywords: i-type lysozyme, Eriocheir sinensis, cloning, characterization

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775 Early Formation of Adipocere in Subtropical Climate

Authors: Asit K. Sikary, O. P. Murty

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Adipocere formation is a modification of the process of putrefaction. It consists mainly of saturated fatty acids, formed by the post-mortem hydrolysis and hydrogenation of body fats with the help of bacterial enzymes in the presence of warmth, moisture and anaerobic bacteria. In temperate climate, it takes weeks to develop while in India it starts to begin within 4-5 days. In this study, we have collected cases with adipocere formation, which were from the South Delhi region (average room temperature 27-390C) and autopsied at our centre. Details of the circumstances of the death, cause and time of death, surrounding environment and demographic profile of the deceased were taken into account. Total 16 cases were included in this study. Adipocere formation was predominantly present over cheeks, shoulder, breast, flanks, buttocks, and thighs. Out of 16, 11 cases were found in a dry atmosphere, 5 cases were brought from the water. There were 5 cases in which adipocere formation was seen in less than 2 days, and among them, in 1 case, as early as one day. This study showed that adipocere formation can be seen as early as 1 day in a hot and humid environment.

Keywords: adipocere, drowning, hanging, humid environment, strangulation, subtropical climate

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774 Prevalence of Methylenetetrahydrofolate Reductase A1298C Variant in Tunisian Childhood Acute Lymphoblastic Leukemia

Authors: Rim Frikha, Maha Ben Jema, Moez Elloumi, Tarek Rebai

Abstract:

Background: Acute lymphoblastic leukemia (ALL); a common blood cancer characterized by the interaction between genetic and environmental factors. Methylenetetrahydrofolate reductase (MTHFR) is an essential folate metabolic enzyme in the processes of DNA synthesis and methylation. A common functional variant of the MTHFR gene, the A1298C, which induces disturbances in folate metabolism, may affect susceptibility to ALL. Objective: The present study aimed to assess the prevalence of MTHFR polymorphism A1298 > C in Tunisian children with ALL. Materials and Methods: A total of 28 Tunisian ALL children were enrolled in this study. Genomic DNA was extracted from whole venous blood collected in ethylenediaminetetraacetic acid (EDTA). Genotyping was carried out with restriction fragment length polymorphism (RFLP) using MboII restriction enzyme. Genotype distribution and allele frequency of MTHFR A1298C was calculated in ALL patients. Results: The A1298C variant of MTHFR was found in 11(19.6%) heterozygous and one homozygous patient (3.5%). Conclusions: This result highlights that A1298C polymorphism of MTHFR is common in Tunisian childhood ALL and suggests that this variant may have a potential role in leukemogenesis. Genotyping of large samples and different ethnicities are required to validate these findings.

Keywords: methylenetetrahydrofolate reductase, acute lymphoblastic leukemia, A1298C variant, prevalence

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773 Cloning and Expression of the ansZ Gene from Bacillus sp. CH11 Isolated from Chilca salterns in Peru

Authors: Stephy Saavedra, Annsy C. Arredondo, Gisele Monteiro, Adalberto Pessoa Jr, Carol N. Flores-Fernandez, Amparo I. Zavaleta

Abstract:

L-asparaginase from bacterial sources is used in leukemic treatment and food industry. This enzyme is classified based on its affinity towards L-asparagine and L-glutamine. Likewise, ansZ genes express L-asparaginase with higher affinity to L-asparagine. The aim of this work was to clone and express of ansZ gene from Bacillus sp. CH11 isolated from Chilca salterns in Peru. The gene encoding L-asparaginase was cloned into pET15b vector and transformed in Escherichia coli BL21 (DE3) pLysS. The expression was carried out in a batch culture using LB broth and 0.5 mM IPTG. The recombinant L-asparaginase showed a molecular weight of ~ 39 kDa by SDS PAGE and a specific activity of 3.19 IU/mg of protein. The cloning and expression of ansZ gene from this halotolerant Bacillus sp. CH11 allowed having a biological input to improve a future scaling-up.

Keywords: ansZ gene, Bacillus sp, Chilca salterns, recombinant L-asparaginase

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772 Composition and in Vitro Antimicrobial Activity of Three Eryngium L. Species

Authors: R. Mickiene, A. Friese, U. Rosler, A. Maruska, O. Ragazinskiene

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This research focuses on phytochemistry and antimicrobial activities of compounds isolated and identified from three species of Eryngium. The antimicrobial activity of extracts from Eryngiumplanum L., Eryngium maritimum L., Eryngium campestre L. grown in Lithuania, were tested by the method of series dilutions, against different bacteria species: Escherichia coli, Proteus vulgaris and Staphylococcus aureus with and without antibiotic resistances, originating from livestock. The antimicrobial activity of extracts was described by determination of the minimal inhibitory concentration. Preliminary results show that the minimal inhibitory concentration range between 8.0 % and 17.0 % for the different Eryngium extracts and bacterial species.The total amounts ofphenolic compounds and total amounts of flavonoids were tested in the methanolic extracts of the plants. Identification and evaluation of the phenolic compounds were performed by liquid chromatography. The essential oils were analyzed by gas chromatography mass spectrometry.

Keywords: antimicrobial activities, Eryngium L. species, essential oils, gas chromatography mass spectrometry

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771 Antibiotic Resistance and Tolerance to Biocides in Enterobacter

Authors: Rebiahi Sid Ahmed, Boutarfi Zakaria, Rahmoun Malika, Antonio Galvez

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The objective of this study was to explore the possible correlation between resistance to antibiotics and tolerance to biocides in Gram-negative bacilli isolated from the University Hospital Center of Tlemcen. This study focused on 175 clinical isolates of Gram-negative bacilli, it is a question of exploring: their level and profile of resistance to antibiotics, their tolerance to biocides, as well as the identification of the genetic supports of this resistance. Enterobacter spp. was the most predominant bacterial genus, all isolates harbored at least one of the studied genes with significant resistance capacity. Our results show, in some cases, a possible positive correlation between the presence of biocide tolerance genes and those of antibiotic resistance; in fact, tolerance to biocides could be one of the co-selection factors for antibiotic resistance. The results of this study should encourage the good practice of hygiene measures as well as the rational use of antimicrobials in order to hinder the development and emergence of resistance in our hospital departments.Mots clés : Antibiotiques, Biocides, Enterobacter, Hôpital, Résistance,

Keywords: antibiotic, biocides, enterobacter, hospital, resistance

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770 Insecticide Resistance Detection on Dengue Vector, Aedes albopictus Obtained from Kapit, Kuching and Sibu Districts in Sarawak State, Malaysia

Authors: Koon Weng Lau, Chee Dhang Chen, Abdul Aziz Azidah, Mohd Sofian-Azirun

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Recently, Sarawak state of Malaysia encounter an outbreak of dengue fever. Aedes albopictus has incriminated as one of the important vectors of dengue transmission. Without an effective vaccine, approaches to control or prevent dengue will be a focus on the vectors. The control of Aedes mosquitoes is still dependent on the use of chemical insecticides and insecticide resistance represents a threat to the effectiveness of vector control. This study was conducted to determine the resistance status of 11 active ingredients representing four major insecticide classes: DDT, dieldrin, malathion, fenitrothion, bendiocarb, propoxur, etofenprox, deltamethrin, lambda-cyhalothrin, cyfluthrin, and permethrin. Standard WHO test procedures were conducted to determine the insecticide susceptibility. Aedes albopictus collected from Kapit (resistance ratio, RR = 1.04–3.02), Kuching (RR = 1.17–4.61), and Sibu (RR = 1.06–3.59) exhibited low resistance toward all insecticides except dieldrin. This study reveled that dieldrin is still effective against Ae. albopictus, followed by fenitrothion, cyfluthrin, and deltamethrin. In conclusion, Ae. albopictus in Sarawak exhibited different resistance levels toward various insecticides and alternative solutions should be implemented to prevent further deterioration of the condition.

Keywords: Aedes albopictus, dengue, insecticide resistance, Malaysia

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769 The Effect of Some Microorganisms from Gastrointestinal Tracts on the Nutritive Value of Broiler Diets

Authors: S. Sangsoponjit, W. Suphalucksana, K. Soytong

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A 2x2 factorial experiment was carried out to determine the effects of two levels of diet supplemented with and without microorganisms in combination with and without feed sterilisation on the nutritive value of broiler diets with four replications in each treatment. Some microorganisms from the gastrointestinal tracts of chickens were supplemented in commercial broiler diets. They were bacterial (BC-NA-01), actinomycetes (BI-NA-03, BC-NA-02 and BL-NA-02), Aspergillus niger sp.(BD-PDA-01), Mucor sp.(BL-PDA-02), Rhizopus stolonifer sp.(BI-PDA-02) and Trichoderma sp.(BL-PDA-02). The results of the proximate analysis revealed that the diet supplemented with microorganisms had a higher percentage of DM and CF in the starter diet(0-3 wks), grower diet(4-5wks) and finisher diet (last period) than the diet without microorganisms (p<0.05). Also, they were higher in the percentage of CP in the starter diet and EE in both the starter diet and grower diet than the diet without microorganisms (p<0.05). The sterilised diet had a higher percentage of moisture than the non-sterilized diet (p<0.01). Also, they were higher in the percentage of CP in the starter diet and CF in both the grower diet and finisher diet than the non-sterilized diet (p<0.05). The sterilized diet supplemented with microorganisms was higher in ME than the non-sterilize diet without microorganisms in the starter diet, grower diet and finisher diet (P<0.01).

Keywords: microorganisms, gastrointestinal tract, nutritive value, broiler diets

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768 Crystallography Trials of Escherichia coli Nitrate Transporter, NarU

Authors: Naureen Akhtar

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The stability of the protein in detergent-containing solution is the key for its successful crystallisation. Fluorescence-detection size-exclusion chromatography (FSEC) is a potential approach for screening monodispersity as well as the stability of protein in a detergent-containing-solution. In this present study, covalently linked Green Fluorescent Protein (GFP) to bacterial nitrate transporter, NarU from Escherichia coli was studied for pre-crystallisation trials by FSEC. Immobilised metal ion affinity chromatography (IMAC) and gel filtration were employed for their purification. The main objectives of this study were over-expression, detergent screening and crystallisation of nitrate transporter proteins. This study could not produce enough proteins that could realistically be taken forward to achieve the objectives set for this particular research. In future work, different combinations of variables like vectors, tags, creation of mutant proteins, host cells, position of GFP (N- or C-terminal) and/or membrane proteins would be tried to determine the best combination as the principle of technique is still promising.

Keywords: transporters, detergents, over-expression, crystallography

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767 Improvement of Antibacterial Activity for Ceftazidime by Partially Purified Tannase from Penicillium expansum

Authors: Sahira N. Muslim, Alaa N. Mohammed, Saba Saadoon Khazaal, Batool Kadham Salman, Israa M. S. AL-Kadmy, Sraa N. Muslim, Ahmed S. Dwaish, Sawsan Mohammed Kareem, Sarah N. Aziz, Ruaa A. Jasim

Abstract:

Tannase has wide applications in food, beverage, brewing, cosmetics and chemical industries and one of the major applications of tannase is the production of gallic acid. Gallic acid is used for manufacturing of trimethoprim. In the present study, a local fungal strain of Penicillium expansum A4 isolated from spoilt apple samples gave the highest production level of tannase. Tannase was partially purified with a recovery yield of 92.52% and 6.32 fold of purification by precipitation using ammonium sulfate at 50% saturation. Tannase led to increased antimicrobial activity of ceftazidime against Pseudomonas aeruginosa and S. aureus and had a synergism effect at low concentrations of ceftazidime, and thus, tannase may be a useful adjuvant agent for the treatment of many bacterial infections in combination with ceftazidime.

Keywords: ceftazidime, Penicillium expansum, tannase, antimicrobial activity

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