Search results for: bacterial aggregation

Authors: Ruta Mickiene, Audrius Maruska, Ona Ragazinskiene

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This research focuses on phytochemistry and antimicrobial activities of compounds isolated and identified from species Rosmarinus officinalis L. This is a study of synergistic effects between phenolic fraction and essential oils. The antimicrobial activity of extracts from Rosmarinus officinalis L. originated from the sector of medicinal plants, Kaunas botanical garden of Vytautas Magnus University Lithuania, were tested by the method of series dilutions, against different bacteria species. Investigated microorganisms were Escherichia coli, Proteus vulgaris and Staphylococcus aureus with and without antibiotic resistances originating from livestock. The antimicrobial activities of extracts were described by determination of the Minimal Inhibitory Concentration (MIC). Preliminary results show that the MIC range between 9.0 % and 12.0 % for the different Rosmarinus officinalis L. extracts and bacterial species. The total amounts of phenolic compounds and total amounts of flavonoids were tested in the methanolic extracts of the plants. The chemical composition for essential oils analysed by GC/MS. Predominant components were alpha pinene (20%), camphor (10%), 1.8‐cineole (5%), phellandrene (6%), camphene (5%), beta pinene (4%), bornylacetate (4%), limonene (2%), borneol (3%), alpha terpineol (3%), cymene (2%), caryophyllene (15%), verbenone (7%), alpha terpinene (3%), eucalyptol (11%).

Keywords: antimicrobial activity, essential oil, Rosmarinus officinalis L., escherichia coli

Procedia PDF Downloads 383

Authors: D. Malouch, M. Berchel, C. Dreanno, S. Stachowski-Haberkorn, P-A. Jaffres

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Biofilm is a predominant lifestyle chosen by bacteria. Whether it is developed on an immerged surface or a mobile biofilm known as flocs, the bacteria within this form of life show properties different from its planktonic ones. Within the biofilm, the self-formed matrix of Extracellular Polymeric Substances (EPS) offers hydration, resources capture, enhanced resistance to antimicrobial agents, and allows cell-communication. Biofouling is a complex natural phenomenon that involves biological, physical and chemical properties related to the environment, the submerged surface and the living organisms involved. Bio-colonization of artificial structures can cause various economic and environmental impacts. The increase in costs associated with the over-consumption of fuel from biocolonized vessels has been widely studied. Measurement drifts from submerged sensors, as well as obstructions in heat exchangers, and deterioration of offshore structures are major difficulties that industries are dealing with. Therefore, surfaces that inhibit biocolonization are required in different areas (water treatment, marine paints, etc.) and many efforts have been devoted to produce efficient and eco-compatible antifouling agents. The different steps of surface fouling are widely described in literature. Studying the biofilm and its stages provides a better understanding of how to elaborate more efficient antifouling strategies. Several approaches are currently applied, such as the use of biocide anti-fouling paint6 (mainly with copper derivatives) and super-hydrophobic coatings. While these two processes are proving to be the most effective, they are not entirely satisfactory, especially in a context of a changing legislation. Nowadays, the challenge is to prevent biofouling with non-biocide compounds, offering a cost effective solution, but with no toxic effects on marine organisms. Since the micro-fouling phase plays an important role in the regulation of the following steps of biofilm formation7, it is desired to reduce or delate biofouling of a given surface by inhibiting the micro fouling at its early stages. In our recent works, we reported that some amphiphilic compounds exhibited bacteriostatic or bactericidal properties at a concentration that did not affect eukaryotic cells. These remarkable properties invited us to assess this type of bio-inspired phospholipids9 to prevent the colonization of surfaces by marine bacteria. Of note, other studies reported that amphiphilic compounds interacted with bacteria leading to a reduction of their development. An amphiphilic compound is a molecule consisting of a hydrophobic domain and a polar head (ionic or non-ionic). These compounds appear to have interesting antifouling properties: some ionic compounds have shown antimicrobial activity, and zwitterions can reduce nonspecific adsorption of proteins. Herein, we investigate the potential of amphiphilic compounds as inhibitors of bacterial growth and marine biofilm formation. The aim of this study is to compare the efficacy of four synthetic phospholipids that features a cationic charge (BSV36, KLN47) or a zwitterionic polar-head group (SL386, MB2871) to prevent microfouling with marine bacteria. We also study the toxicity of these compounds in order to identify the most promising compound that must feature high anti-adhesive properties and a low cytotoxicity on two links representative of coastal marine food webs: phytoplankton and oyster larvae.

Keywords: amphiphilic phospholipids, bacterial biofilm, marine microfouling, non-toxic antifouling

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Authors: Sara Boulmai̇z

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In order to study the microbiological quality of chawarma sold in Biskra, a sampling through some fastfoods of the city was done, the parameters studied are highlighted according to the criteria required by the country's trade management. Microbiological analyzes revealed different levels of contamination by microorganisms. The 10 samples were of an overall view of unsatisfactory quality, and according to the standards, no sample was satisfactory. The range of total aerobic mesophilic flora found is between 105 and 1.2 × 10 7 CFU / g, that of fecal coliforms is 104 to 2.4 × 10 5 CFU / g. The suspected pathogenic staphylococci were between 3.103 and 2.7.106 CFU / g. Salmonellae were absent in all samples, whereas sulphite-reducing anaerobes were present in a single sample. The rate of E. cloacae was between 103 and 6.104 CFU / g. As for fungi and safe mice, their rate was 103 to 107 CFU / g. The study of the sensitivity of antibiotics showed multi-resistance to all the antibiotics tested, although there is a sensitivity towards others. All strains of Staphylococcus aureus tested demonstrated resistance against erythromycin, 30% against streptomycin, and 10% against tetracycline. While the strains of E. cloacae were resistant in all strains to amoxicillin, ceftazidime, cefotaxime, and erythromycin, while they were sensitive to fosfomycin, sulfamethoxazole trimethoperine, ciprofloxacin, and tetracycline. While against chlorophenicol and ofloxacin, the sensitivity was dominant, although there was intermediate resistance. In this study demonstrates that foodborne illnesses remain a problem that arises in addition to the increasingly observed bacterial resistance and that, after all, healthy eating is a right.

Keywords: chawarma, microbiological quality, pathogens., street food

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Authors: Daria Olkiewicz, Maciej Walczak

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In this paper, the biocidal effects of trans-cinnamaldehyde (a main component of cinnamon oil) and geraniol (a constituent of Pelargonium graveolens essential oil) are presented. The activity of the combination of trans-cinnamaldehyde and geraniol was tested against 3 bacterial strains: Staphylococcus aureus ATCC6538 (Gramm+), Escherichia coli ATCC8739 (Gramm-, Lac+) and Pseudomonas aeruginosa KKP 991(Gramm-, Lac-). The biocidal activity of trans-cinnamaldehyde-geraniol mixture against bacteria mentioned above was evaluated by disk-diffusion method. The model strains were exposed on 1, 2.5, 5 and 10 mg of trans-cinnamaldehyde-geraniol mixture per disk, and all strains were susceptible to this combination of plant compounds. For all microorganisms, also Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) were estimated. For Staphylococcus aureus MIC was 0.0625 mg/ml of the trans-cinnamaldehyde and geraniol mixture, and MBC was 1.25 mg/ml; For Escherichia coli MIC=0.5 mg/ml, MBC=1 mg/ml, and finally Pseudomonas aeruginosa was inhibited in 0.5 mg/ml, and minimal biocidal concentration of tested mixture for it was 1.25 mg/ml. There are also reports about the synergistic working of trans-cinnamaldehyde and geraniol against microorganisms and the antimicrobial activity of polymers enriched with trans-cinnamaldehyde or geraniol, therefore the successful development and introduction to the today life of biocidal polymer enriched with trans-cinnamaldehyde and geraniol are possible.

Keywords: antibacterial activity, biocidal polymers, geraniol, trans-cinnamaldehyde

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Authors: Guillaume Lemahieu, Matthias Sentis, Giovanni Brambilla, Gérard Meunier

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Static Multiple Light Scattering (SMLS) has been shown to be a straightforward technique for the characterization of colloidal dispersions without dilution, as multiply scattered light in backscattered and transmitted mode is directly related to the concentration and size of scatterers present in the sample. In this view, the use of SMLS for stability measurement of various dispersion types has already been widely described in the literature. Indeed, starting from a homogeneous dispersion, the variation of backscattered or transmitted light can be attributed to destabilization phenomena, such as migration (sedimentation, creaming) or particle size variation (flocculation, aggregation). In a view to investigating more on the dispersibility of colloidal suspensions, an experimental set-up for “at the line” SMLS experiment has been developed to understand the impact of the formulation parameters on particle size and dispersibility. The SMLS experiment is performed with a high acquisition rate (up to 10 measurements per second), without dilution, and under direct agitation. Using such experimental device, SMLS detection can be combined with the Hansen approach to optimize the dispersing and stabilizing properties of TiO₂ particles. It appears that the dispersibility and the stability spheres generated are clearly separated, arguing that lower stability is not necessarily a consequence of poor dispersibility. Beyond this clarification, this combined SMLS-Hansen approach is a major step toward the optimization of dispersibility and stability of colloidal formulations by finding solvents having the best compromise between dispersing and stabilizing properties. Such study can be intended to find better dispersion media, greener and cheaper solvents to optimize particles suspensions, reduce the content of costly stabilizing additives or satisfy product regulatory requirements evolution in various industrial fields using suspensions (paints & inks, coatings, cosmetics, energy).

Keywords: dispersibility, stability, Hansen parameters, particles, solvents

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Authors: Ali Mohamadi Sani

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The aim of this study was to determine the antimicrobial effect of Helichrysum arenarium L. essential oil in "in-vitro" condition on the growth of seven microbial species including Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Saccharomyces cereviciae, Candida albicans, Aspergillus flavus and Aspergillus parasiticus using microdilution method. The minimum inhibitory concentration (MIC) and minimum bactericidal or fungicidal concentration (MBC, MFC) were determined for the essential oil at ten concentrations. Finally, the sensitivity of tested microbes to the essential oil of H. arenarium was investigated. Results showed that Bacillus subtilis (MIC=781.25 and MBC=6250 µg/ml) was more resistance than two other bacterial species. Among the tested yeasts, Saccharomyces cereviciae (MIC=97.65 and MFC=781.25 µg/ml) was more sensitive than Candida albicans, while among the fungal species, growth of Aspergillus parasiticus inhibited at lower concentration of oil than the Aspergillus flavus. The extracted essential oil exhibited the same MIC value in the liquid medium against all fungal strains (48.82 µg/ml), while different activity against A. flavus and A. parasiticus was observed in this medium with MFC values of 6250 and 390.625µg/ml, respectively. The results of the present study indicated that Helichrysum arenarium L essential oil had significant (P<0.05) antimicrobial activity; therefore, it can be used as a natural preservation to increase the shelf life of food products.

Keywords: Helichrysum arenarium, antimicrobial, essential oil, MIC

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Authors: Asif Jamal, Asad Ali, Muhammad Ishtiaq Ali

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The agriculture productivity losses due to microbial pathogens have been a serious issue in Pakistan and rest of the world. Present work was designed to isolate soil borne microorganisms having the antagonistic ability against notorious phytopathogens. From the initial collection of 23 bacterial isolates, two potent strains of Bacillus were screened on the basis of their comparative efficacy against devastating fungal pathogens. The strains AK-1 and AK-5 showed excellent inhibitory indexes against the majority of tested fungal strains. It was noted that both strains of Bacillus showed significant biocontrolling activity against Aspergillus flavus, Fusarium moniliforme, Colletotricum falcatum, Botrytis cinerea, Aspergillus niger, Fusarium oxysporum, Phythopthora capsici and Rhizopus oryzae. The strain AK-1 was efficient to suppress Aspergillus species and Rhizopus oryzae while AK-5 expressed significant antagonistic activity against Fusarium, Botrytis, and Colletotricum species. On the basis of in vitro assay, it can be postulated that the Bacillus strains AK-1 and AK-5 can be used as a bio-protective agent against various plant diseases. In addition, their applications as natural pesticides could be very helpful to prevent the adverse effects of chemical pesticides.

Keywords: biological control, Bacillus spp, fungal pathogens, agriculture

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Authors: Meksem Nabila, Bordjiba Ouahiba, Meraghni Messaouda, Meksem Amara Leila, Djebar Mohhamed Reda

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The problems of protection of the cultures are being more and more important that they interest great number of farmers and scientists because of the excessive use of the organic phytosanitary products of synthesis that causes fatal damages on the environment. To reduce the inconveniences produced by these pesticides, the use of "biopesticides" originated from plants could be an alternative. The aim of this work is the valuation of a botanical species: Eucalyptus camaldulensis from Northeastern Algeria which extracts are supposed to have an antimicrobial activity, similar to pesticides. The extraction of secondary metabolites from the leaves of E. camaldulensis was realized using methanol and water, and measurements of total polyphenols were made by spectrometric method. Determination of the antimicrobial activity of the extracts at issue was realized in vitro on phyto-pathogenic fungal and bacterial stumps. Tests of comparison were included in the essays by using the chemical pesticidal products of synthesis. The obtained results show that the plant contains polyphenols with an efficiency mattering of the order of 22 %. These polyphenols have a strong fungicidal and bactericidal pesticidal activity against various microbial stumps and the values of the zones of inhibition are more important compared with that obtained in the presence of the chemicals of synthesis (fungicide).

Keywords: eucalyptus camaldulensis, biopesticide, polyphenols, antimicrobial activity

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Authors: Anita Vidacs, Robert Rajko, Csaba Vagvolgyi, Judit Krisch

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With the optimization of a new disinfectant the number of tests could be decreased and the cost of processing too. Good sanitizers are eco-friendly and allow no resistance evolvement of bacteria. The essential oils (EOs) are natural antimicrobials, and most of them have the Generally Recognized As Safe (GRAS) status. In our study, the effect of the EOs cinnamon, marjoram, and thyme was investigated against mixed species bacterial biofilms of Escherichia coli, Listeria monocytogenes, Pseudomonas putida, and Staphylococcus aureus. The optimal concentration of EOs, disinfection time and level of pH were evaluated with the aid of Response Surface Box-Behnken Design (RSD) on 1 day and 7 days old biofilms on metal, plastic, and wood surfaces. The variable factors were in the range of 1-3 times of minimum bactericide concentration (MBC); 10-110 minutes acting time and 4.5- 7.5 pH. The optimized EO disinfectant was compared to industrial used chemicals (HC-DPE, Hypo). The natural based disinfectants were applicable; the acting time was below 30 minutes. EOs were able to eliminate the biofilm from the used surfaces except from wood. The disinfection effect of the EO based natural solutions was in most cases equivalent or better compared to chemical sanitizers used in food industry.

Keywords: biofilm, Box-Behnken design, disinfectant, essential oil

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Authors: Anuja Singh, Avik K. Pati, Ashok K. Mishra

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Organic fluorophores with π-conjugated scaffolds are important because of their interesting optoelectronic properties. In recent years, our lab has been engaged in understanding the photophysics of small diacetylene bridged fluorophores and found the diynes as a promising class of π-conjugated fluorophores. Building on this understanding, recently we have focused on the photophysics of a less explored class of cross-conjugated Y-shaped enediynes (one double and two triple bonds). Here we present the photophysical properties of such enediynes which show interesting photophysical properties that include dual emissions from locally excited (LE) and intramolecular charge transfer (ICT) states and ring size dependent aggregate fluorescence in non-aqueous media. The dyes also show prominent aggregate fluorescence in mixed-aqueous solvents and solid powder form. We further show that the solid state fluorescence can be reversibly switched multiple of cycles by external stimuli, highlighting their potential applications in solid states. The enediynes with push-pull electronic substituents/moieties exhibit high contrast fluorescence color switching upon continuous photon illumination. The intriguing photophysical outcomes of the enediynyl fluorophores are judiciously exploited to generate single-component white light emission in binary solvent mixtures and sense polar aprotic vapor in polymer film matrices. The photophysical behavior of the dyes is further successfully utilized to monitor the microenvironment changes of biologically relevant anisotropic media such as bile salts. In summary, the newly introduced cross-conjugated enediynes enrich the toolbox of organic fluorophores and vouch to display versatile applications.

Keywords: aggregation in solution and solid state, enediynes, physical photochemistry and photophysics, vapor sensing and white light emission

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Authors: Shashi Arya, Nand L. Singh, Samiksha Singh, Pradeep K. Mishra, Siddh N. Upadhyay

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The enormous generation of E- waste and its recycling have greater environmental concern especially in developing countries like India. A major part of this waste comprises printed circuit boards (PCBs). Edge trims of PCBs have high copper content ranging between 25-60%. The extraction of various metals out of these PCBs is more or less a proven technology, wherein various hazardous chemicals are being used in the resource recovery, resulting into secondary pollution. The current trend of extracting of valuable metals is the utilization of microbial strains to eliminate the problem of a secondary pollutant. Keeping the above context in mind, this work aims at the enhanced recovery of copper from edge trims, through bioleaching using bacterial strain Acidithiobacillus ferrooxidans. The raw material such as motherboards, hard drives, floppy drives and DVD drives were obtained from the warehouse of the University. More than 90% copper could be extracted through bioleaching using Acidithiobacillus ferrooxidans. Inoculate concentration has merely insignificant effect over copper recovery above 20% inoculate concentration. Higher concentration of inoculation has the only initial advantage up to 2-4 days. The complete recovery has been obtained between 14- 24 days.

Keywords: acidithiobacillus ferrooxidans, bioleaching, e-waste, printed circuit boards

Procedia PDF Downloads 328

Authors: Sadaf Ilyas

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Recombinant cytokines have been employed successfully as potential therapeutic agent. Some cytokine therapies are already used as a part of clinical practice, ranging from early exploratory trials to well established therapies that have already received approval. Interleukin 15 is a pleiotropic cytokine having multiple roles in peripheral innate and adaptive immune cell function. It regulates the activation, proliferation and maturation of NK cells, T-cells, monocytes/macrophages and granulocytes, and the interactions between them thus acting as a bridge between innate and adaptive immune responses. Unraveling the biology of IL-15 has revealed some interesting surprises that may point toward some of the first therapeutic applications for this cytokine. In this study, the human interleukin 15 gene was isolated, amplified and ligated to a TA vector which was then transfected to a bacterial host, E. coli Top10F’. The sequence of cloned gene was confirmed and it showed 100% homology with the reported sequence. The confirmed gene was then subcloned in pET Expression system to study the IPTG induced expression of IL-15 gene. Positive expression was obtained for number of clones that showed 15 kd band of IL-15 in SDS-PAGE analysis, indicating the successful strain development that can be studied further to assess the potential therapeutic intervention of this cytokine in relevance to human diseases.

Keywords: Interleukin 15, pET expression system, immune therapy, protein purification

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Authors: D. Sengani, N. Potgieter, P. E. L. Mojapelo

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Three bacteria species which comprise of Gram negative and Gram positive microorganisms were isolated and identified on the basis of morpho-cultural study, catalase tests, oxidase tests and biochemical characteristics were found belonging to different genera including Burkholderia cepacia, Pasteurella pneumotropica and Enterococcus faecalis. The main objective of this study was to isolate and identify PCB degrading bacteria from PCB contaminated soils and test them for their degradation ability of PCBs in natural habitat conditions. The results indicated an overall decrease of PCB concentration level with the gradient average ranging from 1.5 to 1.8 respectively. Enterococcus faecalis removed as much as 32% of PCBs in the contaminated soil samples. Whereas Pasteurella pneumotropica could remove 24% of PCBs, Burkholderia cepacia 21% of PCBs and the mixed culture removed 23%. Data showed that the three bacterial strains could tolerate high concentration of PCBs. The results provided the evidence that naturally occurring bacteria in soil contaminated with PCBs have the potential to degrade PCBs. Statistical analysis showed that there was a significant positive correlation between bacteria growth and treatment with a coefficient of (r) =0.1459 and p value < 0.001.

Keywords: bacteria, bioaccumulation, biodegradation, bioremediation, polychlorinated biphenyls

Procedia PDF Downloads 237

Authors: Sutapa Som Chaudhury, Chitrangada Das Mukhopadhyay

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Alzheimer’s disease is a well-known form of dementia since its discovery in 1906. Current Food and Drug Administration approved medications e.g. cholinesterase inhibitors, memantine offer modest symptomatic relief but do not play any role in disease modification or recovery. In last three decades many small molecules, chaperons, synthetic peptides, partial β-secretase enzyme blocker have been tested for the development of a drug against Alzheimer though did not pass the 3rd clinical phase trials. Here in this study, we designed a PEGylated, aminogenic, tripeptidic polymer with two different molecular weights based on the aggregation prone amino acid sequence 17-20 in amyloid beta (Aβ) 1-42. Being conjugated with poly-ethylene glycol (PEG) which self-assembles into hydrophilic nanoparticles, these PEGylated tripeptides constitute a very good drug delivery system crossing the blood brain barrier while the peptide remains protected from proteolytic degradation and non-specific protein interactions. Moreover, being completely aminogenic they would not raise any side effects. These peptide inhibitors were evaluated for their effectiveness against Aβ42 fibrillation at an early stage of oligomer to fibril formation as well as preformed fibril clearance via Thioflavin T (ThT) assay, dynamic light scattering analyses, atomic force microscopy and scanning electron microscopy. The inhibitors were proved to be safe at a higher concentration of 20µM by the reduction assay of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye. Moreover, SHSY5Y neuroblastoma cells have shown a greater survivability when treated with the inhibitors following Aβ42 fibril and oligomer treatment as compared with the control Aβ42 fibril and/or oligomer treated neuroblastoma cells. These make the peptidic inhibitors a promising compound in the aspect of the discovery of alternative medication for Alzheimer’s disease.

Keywords: Alzheimer’s disease, alternative medication, amyloid beta, PEGylated peptide

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Authors: N. Chikhi-Chorfi, L. Arbia, S. Zenia, H.Lounici

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Opuntia ficus-indica belongs to the Cactaceae family. The cactus is mainly cultivated for its fruit (prickly pear) that, eaten after pealing, is sweet and juicy, and rich in nutritional compounds, such as ascorbic acid and polyphenols. Different parts of O. ficus-indica are used in the traditional medicine of several countries: the cladodes are utilized to reduce serum cholesterol level and blood pressure, for treatment of ulcers, rheumatic pain, wounds, fatigue, capillary fragility, and liver conditions. This original study, investigate the effect of polyphenols of O. ficus indica (cactus) cladodes against periodontal bacteria collected from patients with periodontitis. The quantitative analysis of total polyphenols (TPP) was determined with Follin-Ciocalteu method. Different concentrations of extracts of O. ficus indica were tested by the disk method on two bacterial strains: Porphyromonas gingivalis and Prevotella intermedia responsible for periodontal disease. The results showed a good correlation between the concentration of total polyphenols and the antibacterial activity of the extracts of Opuntia ficus indica against P. gingivalis and P. intermedia with R² = 0.94 and R² = 0.90 respectively. This observation suggests that these extracts could be used in the treatment and prevention of periodontitis.

Keywords: periodontal disease, P. gingivalis, P. intermedia, polyphenols, Opuntia ficus indica

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Authors: Ayuko Itsuki, Sachiyo Aburatani

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Soil neutral sugar contents in Kasuga-yama Hill Primeval Forest, which is a World Heritage Site in Nara, Japan consisting of lowland laurel-leaved forest where natural conditions have been preserved for more than 1,000 years, were examined using the two-step hydrolysis to clarify the source of the neutral sugar and relations with the neutral sugar constituted the soil organic matter and the microbial biomass. Samples were selected from the soil (L, F, H and A horizons) surrounding laurel-leaved (BB-1) and Carpinus japonica (BB-2 and PW) trees for analysis. The neutral sugars were one factor of increasing the fungal and bacterial biomass in the laurel-leaved forest soil (BB-1). The more neutral sugar contents in the Cryptomeria japonica forest soil (PW) contributed to the growth of the bacteria and fungi than those of in the Cryptomeria japonica forest soil (BB-2). The neutral sugars had higher correlation with the numbers of bacteria and fungi counted by the dilution plate count method than by the direct microscopic count method. The numbers of fungi had higher correlation with those of bacteria by the dilution plate method.

Keywords: forest soil, neutral sugars, soil organic matter, two-step hydrolysis

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Authors: S. Alqarni, J. C. McCann, A. Palladino, J. J. Loor

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Seven fistulated Holstein cows were used from 3 weeks prepartum to 4 weeks postpartum to determine the relative abundance of 7 different species of ruminal microorganisms. The prepartum diet was based on corn silage. In the postpartum, diet included ground corn, grain by-products, and alfalfa haylage. Ruminal digesta were collected at five times: -14, -7, 10, 20, and 28 days around parturition. Total DNA from ruminal digesta was isolated and real-time quantitative PCR was used to determine the relative abundance of bacterial species. Eubacterium ruminantium and Selenomonas ruminantium were not affected by time (P>0.05). Megasphaera elsdenii and Prevotella bryantii increased significantly postpartum (P<0.001). Conversely, Butyrivibrio proteoclasticus decreased gradually from -14 through 28 days (P<0.001). Fibrobacter succinogenes was affected by time being lowest at day 10 (P=0.02) while Anaerovibrio lipolytica recorded the lowest abundance at -7 d followed by an increase by 20 days postpartum (P<0.001). Overall, these results indicate that changes in diet after parturition affect the abundance of ruminal bacteria, particularly M. elsdenii (a lactate-utilizing bacteria) and P. bryantii (a starch-degrading bacteria) which increased markedly after parturition likely as a consequence of a higher concentrate intake.

Keywords: rumen bacteria, transition cows, rumen metabolism, peripartal period

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Authors: Yi Sun, George Ed Rainger, Steve P. Watson

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Background: Beyond the classical roles in haemostasis and thrombosis, platelets are important in the initiation and development of various thrombo-inflammatory diseases. In atherosclerosis and deep vein thrombosis, for example, platelets bridge monocytes with endothelium and form heterotypic aggregates with monocytes in the circulation. This can alter monocyte phenotype by inducing their activation, stimulating adhesion and migration. These interactions involve cell surface receptor-ligand pairs on both cells. This list is likely incomplete as new interactions of importance to platelet biology are continuing to be discovered as illustrated by our discovery of PEAR-1 binding to FcεR1α. Results: We have developed a highly sensitive avidity-based assay to identify novel extracellular interactions among 126 recombinantly-expressed platelet cell surface and secreted proteins involved in platelet aggregation. In this study, we will use this method to identify novel platelet-monocyte interactions. We aim to identify ligands for orphan receptors and novel partners of well-known proteins. Identified interactions will be studied in preliminary functional assays to demonstrate relevance to the inflammatory processes supporting atherogenesis. Conclusions: Platelet-monocyte interactions are essential for the development of thromboinflammatory disease. Up until relatively recently, technologies only allow us to limit our studies on each individual protein interaction at a single time. These studies propose for the first time to study the cell surface platelet-monocyte interactions in a systematic large-scale approach using a reliable screening method we have developed. If successful, this will likely to identify previously unknown ligands for important receptors that will be investigated in details and also provide a list of novel interactions for the field. This should stimulate studies on developing alternative therapeutic strategies to treat vascular inflammatory disorders such as atherosclerosis, DVT and sepsis and other clinically important inflammatory conditions.

Keywords: membrane proteins, large-scale screening, platelets, recombinant expression

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Authors: P. H. Mfengwana, S. S. Mashele, L. Verschaeve, R. Anthonissen, I. T. Manduna

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Gunnera perpensa is traditionally used mostly by women for the treatment of different gynaecological related conditions due to its proven uterine contractility effects. The uses of this plant include menstrual pain relief, treatment of infertility and promotion of easy labour. However, even though this plant species has been reported to possess numerous medicinal properties, to author’s best knowledge, its safety has not been investigated. Thus, this study was aimed at investigating the genotoxicity and antigenotoxicity of Gunnera perpensa aqueous, methanol and dichloromethane extracts. The in vitro toxicity of the plant extracts was assessed with the neutral red uptake (NRU) test. Genotoxic and antigenotoxic properties of Gunnera perpensa were investigated using high-throughput assays: bacterial Vitotox test and the alkaline comet assay with and without S9 activation on human C3A cells. Ethyl Methanesulfonate (EMS) and 4-nitroquinoline-oxide (4-NQO) were used as positive controls, respectively. All extracts showed toxicity in a dose-dependent manner; however, that does not mean they were all genotoxic. Methanol extract did show genotoxicity with S9 (metabolism) only at the highest concentration of 500 µg/ml due to increased DNA damage observed, however, no genotoxicity was observed from other concentrations. Therefore, the results show that Gunnera perpensa extracts are genotoxic and not safe for human use.

Keywords: antigenotoxicity, comet test, genotoxicity, Gunnera perpensa, vitotox assay

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Authors: Seyed Hassan Moosa-kazemi, Jalal Mohammadi Soleimani, Hassan Vatandoost, Mohammad Hassan Shirazi, Sara Hajikhani, Roonak Bakhtiari, Morteza Akbari, Siamak Hydarzadeh

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Malaria is an infectious disease and considered most important health problems in the southeast of Iran. Iran is elimination malaria phase and new tool need to vector control. Paraterasgenesis is a new way to cut of life cycle of the malaria parasite. In this study, the microflora of the surface and gut of various stages of Anopheles fluviatilis James as one of the important malaria vector was studied using biochemical and molecular techniques during 2013-2014. Twelve bacteria species were found including; Providencia rettgeri, Morganella morganii, Enterobacter aerogenes, Pseudomonas oryzihabitans, Citrobacter braakii، Citrobacter freundii، Aeromonas hydrophila، Klebsiella oxytoca, Citrobacter koseri, Serratia fonticola، Enterobacter sakazakii and Yersinia pseudotuberculosis. The species of Alcaligenes faecalis, Providencia vermicola and Enterobacter hormaechei were identified in various stages of the vector and confirmed by biochemical and molecular techniques. We found Providencia rettgeri proper candidate for paratransgenesis.

Keywords: Anopheles fluviatilis, bacteria, malaria, Paraterasgenesis, Southern Iran

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Authors: Ben Nadler

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Cell adhesion plays a vital role in many cell activities. The motivation to model cell adhesion is to study important biological processes, such as cell spreading, cell aggregation, tissue formation, and cell adhesion, which are very challenging to study by experimental methods alone. This study provides important insight into cell adhesion, which can lead to improve regenerative medicine and tissue formation techniques. In this presentation the biological cells adhesion is mediated by receptors–ligands binding and the diffusivity of the receptor on the cell membrane surface. The ability of receptors to diffuse on the cell membrane surface yields a very unique and complicated adhesion mechanism, which is exclusive to cells. The phospholipid bilayer, which is the main component in the cell membrane, shows fluid-like behavior associated with the molecules’ diffusivity. The biological cell is modeled as a fluid-like membrane with negligible bending stiffness enclosing the cytoplasm fluid. The in-plane mechanical behavior of the cell membrane is assumed to depend only on the area change, which is motivated by the fluidity of the phospholipid bilayer. In addition, the presence of receptors influences on the local mechanical properties of the cell membrane is accounted for by including stress-free area change, which depends on the receptor density. Based on the physical properties of the receptors and ligands the attraction between the receptors and ligands is modeled as a charged-nonpolar which is a noncovalent interaction. Such interaction is a short-range type, which decays fast with distance. The mobility of the receptor on the cell membrane is modeled using the diffusion equation and Fick’s law is used to model the receptor–receptor interactions. The resultant interaction force, which includes receptor–ligand and receptor–receptor interaction, is decomposed into tangential part, which governs the receptor diffusion, and normal part, which governs the cell deformation and adhesion. The formulation of the governing equations and numerical simulations will be presented. Analysis of the adhesion characteristic and properties are discussed. The roles of various thermomechanical properties of the cell, receptors and ligands on the cell adhesion are investigated.

Keywords: cell adhesion, cell membrane, receptor-ligand interaction, receptor diffusion

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Authors: S. L. J. Tan, N. Billa, C. J. Roberts

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Oral delivery of amphotericin B (AmpB) potentially eliminates constraints and side effects associated with intravenous administration, but remains challenging due to the physicochemical properties of the drug such that it results in meagre bioavailability (0.3%). In an advanced formulation, 1) nanostructured lipid carriers (NLC) were formulated as they can accommodate higher levels of cargoes and restrict drug expulsion and 2) a mucoadhesion feature was incorporated so as to impart sluggish transit of the NLC along the gastrointestinal tract and hence, maximize uptake and improve bioavailability of AmpB. The AmpB-loaded NLC formulation was successfully formulated via high shear homogenisation and ultrasonication. A chitosan coating was adsorbed onto the formed NLC. Physical properties of the formulations; particle size, zeta potential, encapsulation efficiency (%EE), aggregation states and mucoadhesion as well as the effect of the variable pH on the integrity of the formulations were examined. The particle size of the freshly prepared AmpB-loaded NLC was 163.1 ± 0.7 nm, with a negative surface charge and remained essentially stable over 120 days. Adsorption of chitosan caused a significant increase in particle size to 348.0 ± 12 nm with the zeta potential change towards positivity. Interestingly, the chitosan-coated AmpB-loaded NLC (ChiAmpB NLC) showed significant decrease in particle size upon storage, suggesting 'anti-Ostwald' ripening effect. AmpB-loaded NLC formulation showed %EE of 94.3 ± 0.02 % and incorporation of chitosan increased the %EE significantly, to 99.3 ± 0.15 %. This suggests that the addition of chitosan renders stability to the NLC formulation, interacting with the anionic segment of the NLC and preventing the drug leakage. AmpB in both NLC and ChiAmpB NLC showed polyaggregation which is the non-toxic conformation. The mucoadhesiveness of the ChiAmpB NLC formulation was observed in both acidic pH (pH 5.8) and near-neutral pH (pH 6.8) conditions as opposed to AmpB-loaded NLC formulation. Hence, the incorporation of chitosan into the NLC formulation did not only impart mucoadhesive property but also protected against the expulsion of AmpB which makes it well-primed as a potential oral delivery system for AmpB.

Keywords: Amphotericin B, mucoadhesion, nanostructured lipid carriers, oral delivery

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Authors: A. Dali-Ali, F. Agag, H. Beldjilali, A. Oukebdane, K. Meddeber, R. Dali-Yahia, N. Midoun

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The ability of Acinetobacter baumannii to develop multiple resistances towards to the majority of antibiotics explains the therapeutic difficulties encountered in severe infections. Furthermore, its persistence in the humid or dry environment promotes cross-contamination in intensive care units. The aim of our study was to describe the epidemiological and bacterial resistance profiles of hospital-acquired infections caused by Acinetobacter baumannii in the intensive care unit of our teaching hospital. During the study period (June 3, 2012 to December 31, 2013), 305 patients having duration of hospitalization equal or more than 48 hours were included in the study. Among these, 36 had developed, at least, one health-care associated infection caused by Acinetobacter baumannii. The rate of infected patients was equal to 11.8% (36/305). The rate of cumulative incidence of hospital-acquired pneumonia was the highest (9.2%) followed by central venous catheter infection (1.3%). Analysis of the various antibiotic resistance profile shows that 93.8% of the strains were resistant to imipenem. The nosocomial infection control committee set up a special program not only to reduce the high rates of incidence of these infections but also to descrease the rate of imipenem resistance.

Keywords: Acinetobacer baumannii, epidemiological profile, hospital acquired infections, intensive care unit

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Authors: Ikram Mohamed Eltayeb Elsiddig, Abdel Khalig Muddather, Hiba Abdel Rahman Ali, Saad Mohamed Hussein Ayoub

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Anogeissusleiocarpus (Combretaceae) is well known for its medicinal uses in African traditional medicine, for treating many human diseases mainly skin diseases and infections.Mycetoma disease is a fungal and/ or bacterial skin infection, mainly cause by Madurella mycetomatis fungus.This study was carried out in vitro to investigate the antifungal activity of Anogeissusleiocarpus leaf extracts against the isolated pathogenicMadurellamycetomatis, by using the NCCLS modified method compared to Ketoconazole standard drug and MTT assay. The bioactive fraction was subjected to chemical analysis implementing different chromatographic analytical methods (TLC, HPLC, and LC-MS/MS). The results showed significance antifungal activity of A. leiocarpus leaf extractsagainst the isolated pathogenicM. mycetomatis, compared to negative and positive controls. The chloroform fraction showed the highest antifungal activity.The chromatographic analysis of the chloroform fraction with the highest activity showed the presence of important bioactive compounds such as ellagic and flavellagic acids derivatives, flavonoids and stilbenoid, which are well known for their antifungal activity.

Keywords: Anogeissus leiocarpus, crude extracts and fractions of Anogeissus leiocarpus, in vitrosusceptibility of Madurella mycetomatis, Madurella mycetomatis

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Authors: Masome Moeni, Roya Abedizadeh, Elham Aram, Hamid Sadeghi-Abandansari, Davood Sabour, Robert Menzel, Ali Hassanpour

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Mitochondria- targeting immunogenic cell death inducers (MT-ICD) have been designed to trigger intrinsic apoptosis signalling pathway in malignant cells and revive the antitumour immune system. MT-ICD inducers have considered to be non-specific, which can deteriorate the ability to initiate mitochondria-selective oxidative stress, causing high toxicity. Iron oxide nanoparticles (IONPs) can be an ideal candidate as vehicles for utilizing in immunotherapy due to their biocompatibility, modifiable surface chemistry, magnetic characteristics and multi-functional applications in single platform. These types of NPs can facilitate a real time imaging which can provide an effective strategy to analyse pharmacokinetic parameters of nano-formula, including blood circulation time, targeted and controlled release at tumour microenvironment. To our knowledge, the conjugation of IONPs with MT-ICD and oxaliplatin (a chemotherapeutic agent used for the treatment of colorectal cancer) for immunotherapy have not been investigated. Herein, IONPs were generated via co-precipitation reaction at high temperatures, followed by coating the colloidal suspension with tetraethyl orthosilicate and 3-aminopropyltriethoxysilane to optimize their bio-compatibility, preventing aggregation and maintaining stability at physiological pH, then functionalized with (3-carboxypropyl) triphenyl phosphonium bromide for mitochondrial delivery. Analytical results demonstrated the successful process of IONPs functionalization. In particular, the colloidal particles of doped IONPs exhibited an excellent stability and dispersibility. The resultant particles were also successfully loaded with the oxaliplatin for an active mitochondrial targeting in immunotherapy, resulting in well-maintained super-paramagnetic characteristics and stable structure of the functionalized IONPs with nanoscale particle sizes.

Keywords: Immunotherapy, mitochondria, cancer, iron oxide nanoparticle

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Authors: Muhammad Raqib Rasul, Tavga Sulaiman Rashid

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Fertilization increases efficiency and obtains better quality of product recovery in agricultural activities. However, fertilizer consumption increased exponentially throughout the world, causing severe environmental problems. Biofertilizers can be a practical approach to minimize chemical fertilizer sources and ultimately develop soil fertility. This study was carried out to isolate, identify and characterize bacteria from medicinal plant (Rumex tuberosus L. and Verbascum sp.) rhizosphere for in vivo screening. 25 bacterial isolates were isolated and several biochemical tests were performed. Two isolates that were positive for most biochemical tests were chosen for the field experiment. The isolates were identified as Go1 Alcaligenes faecalis (Accession No. OP001725) and T11 (Bacillus sp.) based on the 16S rRNA sequence analysis that was compared with related bacteria in GenBank database using MEGA 6.1. For the field trial isolate GO1 and T11 (separately and mixed), NPK as a positive control was used. Both isolates increased plant height, chlorophyll content, number of tubers, and tuber’s weight. The results demonstrated that these two isolates of bacteria can potentially replace with chemical fertilizers for potato production.

Keywords: biofertilizer, Bacillus subtilis, Alcaligenes faecalis, potato tubers, in vivo screening

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Authors: Hajar Zarei, AliAkbar Zarenejadatashgah, Vuk Uskoković, Hiroshi Watabe

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The reactive oxygen species (ROS) generated by radiation in nuclear diagnostic imaging and radiotherapy could damage the structure of the proteins in noncancerous cells surrounding the tumor. The critical factor in many age-related diseases, such as Alzheimer, Parkinson, or Huntington diseases, is the oxidation of proteins by the ROS as molecular triggers of the given pathologies. Our studies by spectroscopic experiments showed doses close to therapeutic ones (1 to 5 Gy) could lead to changes of secondary and tertiary structures in BSA protein macromolecule as a protein model as well as the aggregation of polypeptide chain but without the fragmentation. For this reason, we investigated the radioprotective effects of natural (vitamin C and E) and synthetic materials (CNPs and FIOMPs) on the structural changes in BSA protein induced by gamma irradiation at a therapeutic dose (3Gy). In the presence of both vitamins and synthetic materials, the spectroscopic studies revealed that irradiated BSA was protected from the structural changes caused by ROS, according to in vitro research. The radioprotective property of CNPs and FIOMPs arises from enzyme mimetic activities (catalase, superoxide dismutase, and peroxidase) and their antioxidant capability against hydroxyl radicals. In the case of FIOMPs, a porous structure also leads to increased ROS recombination with each other in the same radiolytic track and subsequently decreased encounters with BSA. The hydrophilicity of vitamin C resulted in the major scavenging of ROS in the solvent, whereas hydrophobic vitamin E localized on the nonpolar patches of the BSA surface, where it did not only neutralize them thanks to the moderate BSA binding constant but also formed a barrier for diffusing ROS. To the best of our knowledge, there has been a persistent lack of studies investigating the radioactive effect of mentioned materials on proteins. Therefore, the results of our studies can open a new widow for application of these common dietary ingredients and new synthetic NPs in improving the safety of radiotherapy.

Keywords: reactive oxygen species, spectroscopy, bovine serum albumin, gamma radiation, radioprotection

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Authors: Babak Sadeghi, Parisa Ghayomipour

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ZnO/Ag nanocomposites coated with polyvinyl chloride (PVC) were prepared by chemical reduction method, for anti-infection biomaterial application. There is a growing interest in attempts in using biomolecular as the templates to grow inorganic nanocomposites in controlled morphology and structure. By optimizing the experiment conditions, we successfully fabricated high yield of ZnO/Ag nanocomposite with full coverage of high-density polyvinyl chloride (PVC) coating. More importantly, ZnO/Ag nanocomposites were shown to significantly inhibit the growth of S. aureus in solution. It was further shown that ZnO/Ag nanocomposites induced thiol depletion that caused death of S. aureus. The coatings were fully characterized using techniques such as scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-ray diffraction (XRD). Most importantly, compared to uncoated metals, the coatings on PVC promoted healthy antibacterial activity. Importantly, compared to ZnO-Ag -uncoated PVC, the ZnO/Ag nanocomposites coated was approximately triplet more effective in preventing bacteria attachment. The result of Thermal Gravimetric Analysis (TGA) indicates that, the ZnO/Ag nanocomposites are chemically stable in the temperature range from 50 to 900 ºC. This result, for the first time, demonstrates the potential of using ZnO/Ag nanocomposites as a coating material for numerous anti-bacterial applications.

Keywords: nanocomposites, antibacterial activity, scanning electron microscopy (SEM), x-ray diffraction (XRD)

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Authors: Małgorzata Tomczyńska, Joanna Saluk-Bijak

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Background: Graves’ disease is a frequent organ-specific autoimmune thyroid disease, which characterized by the presence of different kind autoantibodies, that, in most cases, act as agonists of the thyrotropin receptor, leading to hyperthyroidism. Role of platelets and lymphocytes can be modulated in the pathophysiology of thyroid autoimmune diseases. Interference in the physiology of platelets can lead to enhanced activity of these cells. Activated platelets can bind to circulating lymphocytes and to affect lymphocyte adhesion. Platelets and lymphocytes can regulate mutual functions. Therefore, the activation of T lymphocytes, as well as blood platelets, is associated with the development of inflammation and oxidative stress within the target tissue. The present study was performed to investigate a platelet-lymphocyte relation by assessing the degree of their mutual aggregation in whole blood of patients with Graves’ disease. Also, the purpose of this study was to examine the impact of platelet interaction on lymphocyte migration capacity. Methods: 30 patients with Graves’ disease were recruited in the study. The matched 30 healthy subjects were served as the control group. Immunophenotyping of lymphocytes was carried out by flow cytometry method. A CytoSelect™ Cell Migration Assay Kit was used to evaluate lymphocyte migration and adhesion to blood platelets. Visual assessment of lymphocyte-platelet aggregate morphology was done using confocal microscope after magnetic cell isolation by Miltenyi Biotec. Results: The migration and functional responses of lymphocytes to blood platelets were greater in the group of Graves’ disease patients compared with healthy controls. The group of Graves’ disease patients exhibited a reduced T lymphocyte and a higher B cell count compared with controls. Based on microscopic analysis, more platelet-lymphocyte aggregates were found in patients than in control. Conclusions: Studies have shown that in Graves' disease, lymphocytes show increased platelet affinity, more strongly migrating toward them, and forming mutual cellular conglomerates. This may be due to the increased activation of blood platelets in this disease.

Keywords: blood platelets, cell migration, Graves’ disease, lymphocytes, lymphocyte-platelet aggregates

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Authors: M. Lizzy Madiwani, Ignatious Ncube, Evelyn Madoroba

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This study was designed to determine the distribution of pathogenic bacteria in household raised chickens and study their virulence and antibiotic profiles. For this purpose, 40 chickens were purchased from families in the Capricorn district and sacrificed for sampling. Tissues were cultured on different bacteriological media followed by biotyping using Matrix-assisted Laser Desorption Ionization-time of Flight (MALDI-TOF). Disk diffusion test was performed to determine the antibiotic susceptibility profiles of these bacteria. Out of a total of 160 tissue samples evaluated, E. coli and Salmonella were detected in these tissues. Furthermore, determination of the pathogenic E. coli and Salmonella strains at species level using primer sets that target selected genes of interest in the polymerase chain reaction (PCR) assay was employed. The invA gene, a confirmatory gene of Salmonella was detected in all the Salmonella isolates. The study revealed that there is a high distribution of Salmonella and pathogenic E. coli in these chickens. Therefore, further studies on identification at the species level are highly recommended to provide management and sanitation practices to lower this prevalence. The antimicrobial susceptibly data generated from this study can be a valuable reference to veterinarians for treating bacterial diseases in poultry.

Keywords: antimicrobial, Escherichia coli, pathogens, Salmonella

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