Search results for: bacterial diversity

Authors: M. F. Miah, K. M. A. Zinnah, M. J. Raihan, H. Ali, M. N. Naser

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As genetic diversity is most important for existing, breeding and production of any fish; this study was undertaken for investigating genetic diversity of freshwater mud eel, Monopterus cuchia at population level where three ecological populations such as flooded area of Sylhet (P1), open water of Moulvibazar (P2) and open water of Sunamganj (P3) districts of Bangladesh were considered. Four arbitrary RAPD primers (OPB-12, C0-4, B-03 and OPB-08) were screened and RAPD banding patterns were analyzed among the populations considering 15 individuals of each population. In total 174, 138 and 149 bands were detected in the populations of P1, P2 and P3 respectively; however, each primer revealed less number of bands in each population. 100% polymorphic loci were recorded in P2 and P3 whereas only one monomorphic locus was observed in P1, recorded 97.5% polymorphism. Different genetic parameters such as inter-individual pairwise similarity, genetic distance, Nei genetic similarity, linkage distances, cluster analysis and allelic information, etc. were considered for measuring genetic diversity. The average inter-individual pairwise similarity was recorded 2.98, 1.47 and 1.35 in P1, P2 and P3 respectively. Considering genetic distance analysis, the highest distance 1 was recorded in P2 and P3 and the lowest genetic distance 0.444 was found in P2. The average Nei genetic similarity was observed 0.19, 0.16 and 0.13 in P1, P2 and P3, respectively; however, the average linkage distance was recorded 24.92, 17.14 and 15.28 in P1, P3 and P2 respectively. Based on linkage distance, genetic clusters were generated in three populations where 6 clades and 7 clusters were found in P1, 3 clades and 5 clusters were observed in P2 and 4 clades and 7 clusters were detected in P3. In addition, allelic information was observed where the frequency of p and q alleles were observed 0.093 and 0.907 in P1, 0.076 and 0.924 in P2, 0.074 and 0.926 in P3 respectively. The average gene diversity was observed highest in P2 (0.132) followed by P3 (0.131) and P1 (0.121) respectively.

Keywords: genetic diversity, Monopterus cuchia, population, RAPD, Bangladesh

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Authors: Ashkan Zargar, Ali Taheri Mirghaed, Zein Talal Barakat, Alireza Khosravi, Hamed Paknejad

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With the increase of bacterial resistance to the chemical antibiotics, replacing it with ecofriendly herbal materials and with no adverse effects in the host body is very important. Therefore, in this study, the effect of combined essential oil (Thymus vulgaris-Origanum magorana and Ziziphora clinopodioides) on the growth behavior of Yersinia ruckeri, Aeromonas hydrophila and Lactococcus garvieae was evaluated. The compositions of the herbal essential oils used in this study were determined by gas chromatography-mass spectrometry (GC-MS) while, the investigating of antimicrobial effects was conducted by the agar-disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and bacterial growth curves determination relied on optical density (OD) at 630 nm. The main compounds were thymol (40.60 %) and limonene (15.98 %) for Thymus vulgaris while carvacrol (57.86 %) and thymol (13.54 %) were the major compounds in Origanum magorana. As regards Ziziphora clinopodiodes, α-pinene (22.6 %) and carvacrol (21.1 %) represented the major constituents. Concerning Yersinia ruckeri, disc-diffusion results showed that t.O.z (50 % Origanum majorana) combined essential oil was presented the best inhibition zone (30.66 mm) but it was exhibited no significant differences with other tested commercial antibiotics except oxytetracycline (P <0/05). The inhibitory activity and the bactericidal effect of the t.O.z, unveiled by the MIC= 0.2 μL /mL and MBC= 1.6 μL /mL values, were clearly the best between all combined oils. The growth behaviour of Yersinia ruckeri was affected by this combined essential oil and changes in temperature and pH conditions affected herbal oil performance. As regard Aeromonas hydrophila, its results were so similar to Yersinia ruckeri results and t.O.z (50 % Origanum majorana) was the best between all combined oils (inhibition zone= 26 mm, MIC= 0.4 μL /mL and MBC= 3.2 μL /mL, combined essential oil was affected bacterial growth behavior). Also for Lactococcus garvieae, t.O.z (50 % Origanum majorana) was the best between all combined oils having the best inhibition zone= 20.66 mm, MIC= 0.8 μL /mL and MBC= 1.6 μL /mL and best effect on inhibiting bacterial growth. Combined herbal essential oils have a good and noticeable effect on the growth behavior of pathogenic bacteria in the laboratory, and by continuing research in the host, they may be a suitable alternative to control, prevent and treat diseases caused by these bacteria.

Keywords: bacterial pathogen, herbal medicine, growth behavior, fish

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Authors: Shruti Malviya, Chris Bowler

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Analysis of microbial eukaryotic diversity is fundamental to understanding ecosystems’ structure, biology, and ecology. Diatoms (Stramenopiles, Bacillariophyceae) are one of the most diverse and ecologically prominent groups of phytoplankton. This study was performed to enhance the understanding of global biodiversity patterns and structure of planktonic diatom communities across the world's oceans. We used the metabarcoding data set generated from the biological samples and associated environmental data collected during the Tara Oceans (2009-2013) global circumnavigation covering all major oceanic provinces. A total of ~18 million diatom V9-18S rDNA tags from 126 sampling stations, constituting 631 size-fractionated plankton communities were generated. Using ~250,000 unique diatom metabarcodes, the global diatom distribution and diversity across size classes, genus and ecological niches was assessed. Notably, our analysis revealed: (i) a new estimate of the total number of planktonic diatom species, (ii) a considerable unknown diversity and exceptionally high diversity in the open ocean, and (iii) complex diversity patterns across oceanic provinces. Also, co-occurrence of several ribotypes in locations separated by great geographic distances (equatorial stations) demonstrated a widespread but not ubiquitous distribution. This work provides a comprehensive perspective on diatom distribution and diversity in the world’s oceans and elaborates interconnections between associated theories and underlying drivers. It shows how meta-barcoding approaches can provide a framework to investigate environmental diversity at a global scale, which is deemed as an essential step in answering various ecological research questions. Consequently, this work also provides a reference point to explore how microbial communities will respond to environmental conditions.

Keywords: diatoms, Tara Oceans, biodiversity, metabarcoding

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Authors: Ayesha Aihetasham, Najma Arshad, Sobia Khan

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Damage causes by subterranean termites are of major concern today. Termites majorly treated with pesticides resulted in several problems related to health and environment. For this reason, plant-derived natural products specifically essential oils have been evaluated in order to control termites. The aim of the present study was to investigate the antitermitic potential of six essential oils on Heterotermes indicola subterranean termite. No-choice bioassay was used to assess the termiticidal action of essential oils. Further, gut from each set of treated termite group was extracted and analyzed for reduction in number of protozoa and bacteria by protozoal count method using haemocytometer and viable bacterial plate count (dilution method) respectively. In no-choice bioassay it was found that Foeniculum vulgare oil causes high degree of mortality 90 % average mortality at 10 mg oil concentration (10mg/0.42g weight of filter paper). Least mortality appeared to be due to Citrus sinensis oil (43.33 % average mortality at 10 mg/0.42g). The highest activity verified to be of Foeniculum vulgare followed by Eruca sativa, Trigonella foenum-graecum, Peganum harmala, Syzygium cumini and Citrus sinensis. The essential oil which caused maximum reduction in number of protozoa was P. harmala followed by T. foenum-graecum and E. sativa. In case of bacterial count E. sativa oil indicated maximum decrease in bacterial number (6.4×10⁹ CFU/ml). It is concluded that F. vulgare, E. sativa and P. harmala essential oils are highly effective against H. indicola termite and its gut microflora.

Keywords: bacterial count, essential oils, Heterotermes indicola, protozoal count

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Authors: Abdul Matin, Muazzama Akhtar, Shahid Nisar, Saddaf Mazzar, Umer Rashid

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Antibiotic resistance is an increasing concern worldwide now a day. Neisseria gonorrhea and Staphylococcus aureus are known to cause major human sexually transmitted and respiratory diseases respectively. Nanotechnology is an emerging discipline and its application in various fields especially in medical sciences is gigantic. In the present study, we synthesized multi-walled carbon nanotubes (MWNTs) using acid oxidation method and solubilized MWNTs were with length predominantly >500 nm and diameters ranging from 40 to 50 nm. The locally synthesized MWNTs were used against gram positive and negative bacteria to determine their impact on bacterial growth. Clinical isolates of Neisseria gonorrhea (isolate: 4C-11) and Staphylococcus aureus (isolate: 38541) were obtained from local hospital and normally cultured in LB broth at 37°C. Both clinical strains can be obtained on request from University of Gujarat. Spectophometric assay was performed to determine the impact of MWNTs on bacterial growth in vitro. To determine the effect of MWTNs on test organisms, various concentration of MWNTs were used and recorded observation on various time intervals to understand the growth inhibition pattern. Our results demonstrated that MWNTs exhibited toxic effects to Staphylococcus aureus while showed very limited growth inhibition to Neisseria gonorrhea, which suggests the resistant potential of Neisseria against nanoparticles. Our results clearly demonstrate the gradual decrease in bacterial numbers with passage of time when compared with control. Maximum bacterial inhibition was observed at maximum concentration (50 µg/ml). Our future work will include further characterization and mode of action of our locally synthesized MWNTs. In conclusion, we investigated and reported for the first time the inhibitory potential of locally synthesized MWNTs on local clinical isolates of Staphylococcus aureus and Neisseria gonorrhea.

Keywords: antibacterial activity, multi walled carbon nanotubes, Neisseria gonorrhea, spectrophotometer assay, Staphylococcus aureus

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Authors: Kubilay Kurtulus Bastas

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Bacterial diseases are very destructive and cause economic losses on pears. Promising plant extracts for the management of plant diseases are environmentally safe, long-lasting and extracts of certain plants contain alkaloids, tannins, quinones, coumarins, phenolic compounds, and phytoalexins. In this study, bacteria were isolated from different parts of pear exhibiting characteristic symptoms of bacterial diseases from the Central Anatolia, Turkey. Pathogenic bacteria were identified by morphological, physiological, biochemical and molecular methods as fire blight (Erwinia amylovora (39%)), bacterial blossom blast and blister bark (Pseudomonas syringae pv. syringae (22%)), crown gall (Rhizobium radiobacter (1%)) from different pear cultivars, and determined virulence levels of the pathogens with pathogenicity tests. The air-dried 25 plant material was ground into fine powder and extraction was performed at room temperature by maceration with 80% (v/v) methanol/distilled water. The minimum inhibitory concentration (MIC) values were determined by using modified disc diffusion method at five different concentrations and streptomycin sulphate was used as control chemical. Bacterial suspensions were prepared as 108 CFU ml⁻¹ densities and 100 µl bacterial suspensions were spread to TSA medium. Antimicrobial activity was evaluated by measuring the inhibition zones in reference to the test organisms. Among the tested plants, Origanum vulgare, Hedera helix, Satureja hortensis, Rhus coriaria, Eucalyptus globulus, Rosmarinus officinalis, Ocimum basilicum, Salvia officinalis, Cuminum cyminum and Thymus vulgaris showed a good antibacterial activity and they inhibited the growth of the pathogens with inhibition zone diameter ranging from 7 to 27 mm at 20% (w/v) in absolute methanol in vitro conditions. In vivo, the highest efficacy was determined as 27% on reducing tumor formation of R. radiobacter, and 48% and 41% on reducing shoot blight of E. amylovora and P. s. pv. syringae on pear seedlings, respectively. Obtaining data indicated that some plant extracts may be used against the bacterial diseases on pome fruits within sustainable and organic management programs.

Keywords: bacteria, eco-friendly management, organic, pear, plant extract

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Authors: T. Deepa, S. R. Inchara, S. V. Venkatesh, Seema Tharannum

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Concrete is the most widely used structural material. It is made using locally available materials. However, Concrete has low tensile strength and may crack in the early days with exothermic hydration. Bacillus subtilis bacteria that form endospores is the biological agent considered in this study for Biomineralization or MICP (Microbially Induced Calcite Precipitation) Technique and to address the increased Construction water demand, Recycled Grey Water which is obtained from STP of PES University, opted in place of Potable water. In this work, M30 grade conventional concrete is designed using OPC 53 grade cement, Manufactured Sand, Natural coarse aggregates, and Potable water. Conventional Concrete (CC), Bacterial Concrete with Potable water (BS), and Recycled Grey Water concrete (RGW) are the three different concrete specimens casted. Experimental studies such as the strength test and the surface hardness test are conducted on Conventional and Bacterial concrete samples after 7, 28, and 56 days of curing. Concrete cubes are subjected to a temperature of 50° C to investigate the effect of higher temperature. Cracked cube specimens are observed for Self-healing - as well as microstructure analysis with Scanning Electron Microscope (SEM), Energy Dispersive X-Ray Analysis (EDAX), and X-Ray Diffraction Analysis (XRD).Noticeable Calcium salt deposition is observed on the surface of BS and RGW cracked specimen. Surface hardness and EDAX test gave promising result on the advantage of using spore-forming bacteria in concrete. This is followed by the strength gain in Compression and Flexure. Results also indicate that Recycled Grey Water can be a substitute for Normal water in concrete.

Keywords: bacillus subtilis, bacterial concrete, recycled grey water, self-healing, surface hardness of concrete

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Authors: Bedilu Tafesse

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Home gardens are important social and cultural spaces where knowledge related to agricultural practice is transmitted and through which households may improve their income and livelihood. High levels of inter- and intra-specific plant genetic diversity are preserved in home gardens. Plant diversity is threatened by rapid and unplanned urbanization, which increases environmental problems such as heating, pollution, loss of habitats and ecosystem disruption. Tropical home gardens have played a significant role in conserving plant diversity while providing substantial benefits to households. This research aimed to understand the relationship between household characteristics and plant diversity in western Ethiopia home gardens and the contributions of plants to the household economy. Plant diversity and different uses of plants were studied in a random sample of 111 suburban home gardens in the Ilu Ababora, Jima and Wellega suburban area, western Ethiopia, based on complete garden inventories followed by household surveys on socio-economic status during 2012. A total of 261 species of plants were observed, of which 41% were ornamental plants, 36% food plants, and 22% medicinal plants. Of these 16% were sold commercially to produce income. Avocado, bananas, and other fruits produced in excess. Home gardens contributed the equivalent of 7% of total annual household income in terms of food and commercial sales. Multiple regression analysis showed that education, time spent in gardening, land for cultivation, household expenses, primary conservation practices, and uses of special techniques explained 56% of the total plant diversity. Food, medicinal and commercial plant species had significant positive relationships with time spent gardening and land area for gardening. Education and conservation practices significantly affected food and medicinal plant diversity. Special techniques used in gardening showed significant positive relations with ornamental and commercial plants. Reassessments in different suburban and urban home gardens and proper documentation using same methodology is essential to build a firm policy for enhancing plant diversity and related values to households and surroundings.

Keywords: plant genetic diversity, urbanization, suburban home gardens, Ethiopia

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Authors: Sadie Namani, Lindita Ajazaj, Arjeta Zogaj, Vera Berisha, Bahrije Halili, Luljeta Hasani, Ajete Aliu

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Background: The outcome of bacterial meningitis is strongly related to the resistance of bacterial pathogens to the initial antimicrobial therapy. The objective of the study was to analyze the initial antimicrobial therapy, the resistance of meningeal pathogens and the outcome of adults bacterial meningitis cases. Materials/methods: This prospective study enrolled 46 adults older than 16 years of age, treated for bacterial meningitis during the years 2009 and 2010 at the infectious diseases clinic in Prishtinë. Patients are categorized into specific age groups: > 16-26 years of age (10 patients), > 26-60 years of age (25 patients) and > 60 years of age (11 patients). All p-values < 0.05 were considered statistically significant. Data were analyzed using Stata 7.1 and SPSS 13. Results: During the two year study period 46 patients (28 males) were treated for bacterial meningitis. 33 patients (72%) had a confirmed bacterial etiology; 13 meningococci, 11 pneumococci, 7 gram-negative bacilli (Ps. aeruginosa 2, Proteus sp. 2, Acinetobacter sp. 2 and Klebsiella sp. 1 case) and 2 staphylococci isolates were found. Neurological complications developed in 17 patients (37%) and the overall mortality rate was 13% (6 deaths). Neurological complications observed were: cerebral abscess (7/46; 15.2%), cerebral edema (4/46; 8.7%); haemiparesis (3/46; 6.5%); recurrent seizures (2/46; 4.3%), and single cases of thrombosis sinus cavernosus, facial nerve palsy and decerebration (1/46; 2.1%). The most common meningeal pathogens were meningococcus in the youngest age group, gram negative-bacilli in second age group and pneumococcus in eldery age group. Initial single-agent antibiotic therapy (ceftriaxone) was used in 17 patients (37%): in 60% of patients in the youngest age group and in 44% of cases in the second age group. 29 patients (63%) were treated with initial dual-agent antibiotic therapy; ceftriaxone in combination with vancomycin or ampicillin. Ceftriaxone and ampicillin were the most commonly used antibiotics for the initial empirical therapy in adults > 50 years of age. All adults > 60 years of age were treated with the initial dual-agent antibiotic therapy as in this age group was recorded the highest mortality rate (M=27%) and adverse outcome (64%). Resistance of pathogens to antimicrobics was recorded in cases caused by gram-negative bacilli and was associated with greater risk for developing neurological complications (p=0.09). None of the gram-negative bacilli were resistant to carbapenems; all were resistant to ampicillin while 5/7 isolates were resistant to cefalosporins. Resistance of meningococci and pneumococci to beta-lactams was not recorded. There were no statistical differences in the occurrence of neurological complications (p > 0.05), resistance of meningeal pathogens to antimicrobics (p > 0.05) and the inital antimicrobial therapy (one vs. two antibiotics) concerning group-ages in adults. Conclusions: The initial antibiotic therapy with ceftriaxone alone or in combination with vancomycin or ampicillin did not cover cases caused by gram-negative bacilli.

Keywords: adults, bacterial meningitis, outcomes, therapy

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Authors: Susan Mwai, Mary Muchane, Peter Wachira, Sheila Okoth, Muchai Muchane, Halima Saado

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Tropical forests harbor a wide range of biodiversity and rich macro-fungi diversity compared to the temperate regions in the World. However, biodiversity is facing the threat of extinction following the rate of forest loss taking place before proper study and documentation of macrofungi is achieved. The present study was undertaken to determine the effect of converting indigenous habitat to plantation forest on macrofungi diversity. To achieve the objective of this study, an inventory focusing on macro-fungi diversity was conducted within Kereita block in Kikuyu Escarpment forest which is on the southern side of Aberdare mountain range. The macrofungi diversity was conducted in the indigenous forest and in more than 15 year old Patula plantation forest , during the wet (long rain season, December 2014) and dry (Short rain season, May, 2015). In each forest type, 15 permanent (20m x 20m) sampling plots distributed across three (3) forest blocks were used. Both field and laboratory methods involved recording abundance of fruiting bodies, taxonomic identity of species and analysis of diversity indices and measures in terms of species richness, density and diversity. R statistical program was used to analyze for species diversity and Canoco 4.5 software for species composition. A total number of 76 genera in 28 families and 224 species were encountered in both forest types. The most represented taxa belonged to the Agaricaceae (16%), Polyporaceae (12%), Marasmiaceae, Mycenaceae (7%) families respectively. Most of the recorded macro-fungi were saprophytic, mostly colonizing the litter 38% and wood 34% based substrates, which was followed by soil organic dwelling species (17%). Ecto-mycorrhiza fungi (5%) and parasitic fungi (2%) were the least encountered. The data established that indigenous forests (native ecosystems) hosts a wide range of macrofungi assemblage in terms of density (2.6 individual fruit bodies / m2), species richness (8.3 species / plot) and species diversity (1.49/ plot level) compared to the plantation forest. The Conversion of native forest to plantation forest also interfered with species composition though did not alter species diversity. Seasonality was also shown to significantly affect the diversity of macro-fungi and 61% of the total species being present during the wet season. Based on the present findings, forested ecosystems in Kenya hold diverse macro-fungi community which warrants conservation measures.

Keywords: diversity, Indigenous forest, macro-fungi, plantation forest, season

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Authors: Sunil M. Gaikwad, Yogesh J. Koli, Gopal A. Raut, Ganesh P. Bhawane

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The present investigation was directed to study the diversity of short-horned grasshoppers from a forested area of Kolhapur district, Maharashtra, India, which is spread along the hilly terrain of the Northern Western Ghats. The collection was made during 2013 to 2015, and identified with the help of a reference collection of ZSI, Kolkata, and recent literature and dry preserved. The study resulted in the enumeration of 40 species of short-horned grasshoppers belonging to four families of suborder: Caelifera. The family Acrididae was dominant (27 species) followed by Tetrigidae (eight species), Pyrgomorphidae (four species) and Chorotypidae (one species). The report of 40 species from the forest habitat of the study region highlights the significance of the Western Ghats. Ecologically, short-horned grasshoppers are integral to food chains, being consumed by a wide variety of animals. The observations of the present investigation may prove useful for conservation of the Diversity in Northern Western Ghats.

Keywords: diversity, Kolhapur, northern western Ghats, short-horned grasshoppers

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Authors: Nada Azhar

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As a developing country, The Kingdom of Saudi Arabia (KSA) needs to make the best possible use of its workforce for social and economic reasons. The workforce is diverse, calling for appropriate diversity management (DM). The thesis focuses on the banking sector in KSA. To date, there have been no studies on DM in the banking sector in this country. Many organizations have introduced specific policies and programmes to improve the recruitment, inclusion, promotion, and retention of diverse employees, in addition to the legal requirements existing in many countries. However, Western-centric models of DM may not be applicable, at least not in their entirety, in other regions. The aim of the study is to devise a framework for understanding gender, age and disability DM in the banking sector in KSA in order to enhance DM in this sector. A sample of 24 managers, 2 from each of the 12 banks, was interviewed to obtain their views on DM in the banking sector in KSA. Thematic analysis was used to analyze the data. These themes were used to develop the questionnaire, which was administered to 10 managers in each of the 12 banks. After analysis of these data, and completion of the study, the research will make a theoretical contribution to the knowledge on DM and a practical contribution to the management of diversity in Saudi banks. This paper concerns a work in progress.

Keywords: age, disability, diversity, gender, Kingdom of Saudi Arabia

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Authors: Wisam H. Benamer, Ehab A. Elfallah, Mohamed A. Elshaari, Farag A. Elshaari

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A challenge for laboratories is to provide bacterial identification and antibiotic sensitivity results within a short time. Hence, advancement in the required technology is desirable to improve timing, accuracy and quality. Even with the current advances in methods used for both phenotypic and genotypic identification of bacteria the need is there to develop method(s) that enhance the outcome of bacteriology laboratories in accuracy and time. The hypothesis introduced here is based on the assumption that the chromosome of any bacteria contains unique sequences that can be used for its identification and typing. The outcome of a pilot study designed to test this hypothesis is reported in this manuscript. Methods: The complete chromosome sequences of several bacterial species were downloaded to use as search targets for unique sequences. Visual basic and SQL server (2014) were used to generate a complete set of 18-base long primers, a process started with reverse translation of randomly chosen 6 amino acids to limit the number of the generated primers. In addition, the software used to scan the downloaded chromosomes using the generated primers for similarities was designed, and the resulting hits were classified according to the number of similar chromosomal sequences, i.e., unique or otherwise. Results: All primers that had identical/similar sequences in the selected genome sequence(s) were classified according to the number of hits in the chromosomes search. Those that were identical to a single site on a single bacterial chromosome were referred to as unique. On the other hand, most generated primers sequences were identical to multiple sites on a single or multiple chromosomes. Following scanning, the generated primers were classified based on ability to differentiate between medically important bacterial and the initial results looks promising. Conclusion: A simple strategy that started by generating primers was introduced; the primers were used to screen bacterial genomes for match. Primer(s) that were uniquely identical to specific DNA sequence on a specific bacterial chromosome were selected. The identified unique sequence can be used in different molecular diagnostic techniques, possibly to identify bacteria. In addition, a single primer that can identify multiple sites in a single chromosome can be exploited for region or genome identification. Although genomes sequences draft of isolates of organism DNA enable high throughput primer design using alignment strategy, and this enhances diagnostic performance in comparison to traditional molecular assays. In this method the generated primers can be used to identify an organism before the draft sequence is completed. In addition, the generated primers can be used to build a bank for easy access of the primers that can be used to identify bacteria.

Keywords: bacteria chromosome, bacterial identification, sequence, primer generation

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Authors: Saqib Saleem Abdullah, Saima Masood, Hafsa Zaneb, Shela Gul Bokhari, Muti Ur Rehman, Jamil Akbar

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A 5- weeks feeding trial was carried out to determine the effectiveness of Bacterial Phytase (Phyzyme®) in broilers, at different dietary levels of Phosphorous. 140 d-old broilers (Hubbard) were randomly divided into 4 groups (n=4). Birds were fed corn-based basal diet or the same diet supplemented with 3 different levels of non Phytate Phosphorous (NPP) (0.45 %, 0.30 % and 0.15 %). Furthermore, the diets were supplemented with bacterial Phytase. Birds were fed ad libitum and kept under thermo neutral conditions. The parameters studied were; body weight gain (BWG), tibial bone characteristics (TBC), serum Calcium (Ca), Phosphorus (P) and Alkaline Phosphatase (AP) levels and tibia ash percentage (TAP). BWG of the broilers was calculated at weekly interval and remaining parameters were calculated after slaughtering the birds at 35thday. Results suggested that Phytase supplementation at 0.30% NPP (Non Phytate Phosphorus + Bacterial Phytase) increased (P < 0.05) the BWG, bone length, bone weight, tibiotarsal index, medullary canal diameter and diaphysis diameter however, rubosticity index was reduced to minimum (P < 0.05) at this dietary level of phosphorous when compared with other groups. Maximum (P < 0.05) rubosticity index was observed in control group with 0% Phytase. Furthermore, Phytase addition at 0.30 % NPP also improved (P < 0.05) Ca, P and AP levels in the blood. Phytase supplementation at lower phosphorus level (0.30%NPP) improved BWG and TBC including bone density and bone quality in broilers hence it can be concluded that addition of Phytase at 0.30% NPP may prove beneficial for bone and overall performance in broilers.

Keywords: diaphysis diameter, phytase, rubosticity index, tibia

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Authors: Carlos R. Duharte Rodríguez, Ibrain Ceballo Acosta, Carmen B. Busoch Morlán, Angel Regueiro Gómez, Annet Martinez Hernández

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This work shows the designing and characterization of a prototype of laboratory incubator as support of research in Microbiology, in particular during studies of bacterial growth in biological samples, with the help of optic methods (Turbidimetry) and electrometric measurements of bioimpedance. It shows the results of simulation and experimentation of the design proposed for the canals of measurement of the variables: temperature and humidity, with a high linearity from the adequate selection of sensors and analogue components of every channel, controlled with help of a microcontroller AT89C51 (ATMEL) with adequate benefits for this type of application.

Keywords: microbiology, bacterial growth, incubation station, microorganisms

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Authors: Samia Dahmani

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The research investigated the process and outcomes of an ongoing project “Den Grønne Rute”: a network of recreational rooms to be implemented throughout a former vulnerable neighborhood: Trekanten in the city of Holstebro in Denmark. The aim of the project is to better integrate Trekanten in the city and which initiated reflections upon the (dis)connection and the integration in relation to urban planning and city management, as well as the extent to which the project considers diversity since Holstebro is a growing multicultural city. With this research, it was first verified if Trekanten is splintered from Holstebro city, and secondly if planning for diversity, by engaging people in the process via place-making approaches, can help redress the disconnection between the neighborhood and the rest of the city. More specifically, the paper aims at exploring the role of place-making “Den Grønne Rute” in Trekanten in promoting planning for diversity and reassembling the splintering in the city. The theoretical and conceptual framework served to analyze the relationship between the splintering urbanism concept and the community involvement’s role in an inclusive process. The field study examines the detachment between Trekanten and Holstebro and the extent to which the project can overcome the disconnection. Methodologically a mix-methods approach was adopted where two semi-structured interviews, a focus group, and an online survey were conducted. Contrary to prior assumptions, the results showed that not only is Trekanten splintered from the city, but also the city is greatly disconnected from Trekanten, and hence Holstebro is a splintered city. The surprise was that Trekanten is moreover inner-splintered. The splintering urbanism accordingly has different dimensions. Even though the project’s design seemed to incorporate diverse ages and groups of people, its process lacks an understanding of the diversity’s relevance in promoting inclusiveness. In fact, the analysis revealed socio-cultural and psychological splintering. Since place-making, as a collaborative approach in planning, is itself an expression of diversity (since it brings differences into play), reconsidering diversity within the process by engaging people at the early stages of planning was recommended. Another suggestion was not to limit the project to a destination but more as an experience to remember and a story to tell. Only by bringing people together in re-imagining the place can Trekanten reassemble with Holstebro and vice versa. The aim of the research was to add a new perspective to the splintering urbanism and planning for diversity so to advance place-making as an approach in promoting the latter and redressing the former.

Keywords: the splintering urbanism, place-making, planning for diversity, Den Grønne Rute, Trekanten, Holstebro

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Authors: Samia Dahmani

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The research investigated the process and outcomes of an ongoing project, “Den Grønne Rute”: a network of recreational rooms to be implemented throughout a former vulnerable neighborhood: Trekanten, in the city of Holstebro in Denmark. The aim of the project is to better integrate Trekanten in the city and which initiated reflections upon the (dis)connection and the integration in relation to urban planning and city management, as well as the extent to which the project considers diversity since Holstebro is a growing multicultural city. With this research, it was first verified if Trekanten is splintered from Holstebro city, and secondly if planning for diversity, by engaging people in the process via place-making approaches, can help redress the disconnection between the neighborhood and the rest of the city. More specifically, the paper aims at exploring the role of place-making “Den Grønne Rute” in Trekanten in promoting planning for diversity and reassembling the splintering in the city. The theoretical and conceptual framework served to analyze the relationship between the splintering urbanism concept and the community involvement’s role for an inclusive process. The field study examines the detachment between Trekanten and Holstebro and the extent to which the project can overcome the disconnection. Methodologically a mix-methods approach was adopted where two semi-structured interviews, a focus group, and an online survey were conducted. Contrary to prior assumptions, the results showed that not only is Trekanten splintered from the city, but also the city is greatly disconnected from Trekanten, and hence Holstebro is a splintered city. The surprise was that Trekanten is moreover inner-splintered. The splintering urbanism accordingly has different dimensions. Even though the project’s design seemed to incorporate diverse ages and groups of people, its process lacks an understanding of the diversity’s relevance in promoting inclusiveness. In fact, the analysis revealed socio-cultural and psychological splintering. Since place-making, as a collaborative approach in planning, is itself an expression of diversity (since it brings differences into play), reconsidering diversity within the process by engaging people at the early sages of planning was recommended. Another suggestion was not to limit the project to a destination but more as an experience to remember and a story to tell. Only by bringing people together in re-imagining the place, can Trekanten reassemble with Holstebro and vice versa. The aim with the research was to add a new perspective to the splintering urbanism and planning for diversity so to advance place-making as an approach in promoting the latter and redressing the former.

Keywords: the splintering urbanism, placemaking, planning for diversity, den grønne rute, trekanten, holstebro

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Authors: O. Baghriche, A. Zertal, C. Pulgarin, J. Kiwi, R. Sanjines

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High-Power Impulse Magnetron Sputtering (HIPIMS) is a technology that belongs to the field of Ionized PVD of thin films. This study shows the first complete report on ultrathin TiO2/Ag nano-particulate films sputtered by highly ionized pulsed plasma magnetron sputtering (HIPIMS) leading to fast bacterial loss of viability. The Ag and the TiO2/Ag sputtered films induced complete Escherichia coli inactivation in the dark, which was not observed in the case of TiO2. When Ag was present, the bacterial inactivation was accelerated under low intensity solar simulated light and this has implications for a potential for a practical technology. The design, preparation, testing and surface characterization of these innovative films are described in this study. The HIPIMS sputtered composite films present an appreciable savings in metals compared to films obtained by conventional sputtering methods. HIPIMS sputtering induces a strong interaction with the rugous polyester 3-D structure due to the higher fraction of the Ag-ions (M+) attained in the magnetron chamber. The immiscibility of Ag and TiO2 in the TiO2/Ag films is shown by High Angular Dark Field (HAADF) microscopy. The ionization degree of the film forming species is significantly increased and film growth is assisted by an intense ion flux. Reports have revealed the significant enhancement of the film properties as the HIPIMS technology is used. However, a decrease of the deposition rate, as compared to the conventional DC magnetron sputtering Pulsed (DCMSP) process is commonly observed during HIPIMS.

Keywords: E. coli, HIPIMS, inactivation bacterial, sputtering

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Authors: Bintou Jobe

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Background: The research is focused on the concept of equality, diversity, and inclusion (EDI) and the need to achieve equity by treating individuals according to their circumstances and needs. The research is rooted in the UK Equality Act 2010, which emphasizes the importance of equal opportunities for all individuals regardless of their background and social life. However, inequality persists in society, particularly for those from minority backgrounds who face discrimination. Research Aim: The aim of this research is to promote equality, diversity, and inclusion by encouraging the regeneration of minds and the eradication of stereotypes. The focus is on promoting good Equality, Diversity and Inclusion practices in various settings, including schools, colleges, universities, and workplaces, to create environments where every individual feels a sense of belonging. Methodology: The research utilises a literature review approach to gather information on promoting inclusivity, diversity, and inclusion. Findings: The research highlights the significance of promoting equality, diversity, and inclusion practices to ensure that individuals receive the respect and dignity they deserve. It emphasises the importance of treating individuals based on their unique circumstances and needs rather than relying on stereotypes. The research also emphasises the benefits of diversity and inclusion in enhancing innovation, creativity, and productivity. The theoretical importance of this research is to raise awareness about the importance of regenerating minds, challenging stereotypes, and promoting equality, diversity, and inclusion. The emphasis is on treating individuals based on their circumstances and needs rather than relying on generalizations. Diversity and inclusion are beneficial in different settings, as highlighted by the research. By raising awareness about the importance of mind regeneration, eradicating stereotypes, and promoting equality, diversity, and inclusion, this research makes a significant contribution to the subject area. It emphasizes the necessity of treating individuals based on their unique circumstances instead of relying on generalizations. However, the methodology could be strengthened by incorporating primary research to complement the literature review approach. Data Collection and Analysis Procedures: The research utilised a literature review approach to gather relevant information on promoting inclusivity, diversity, and inclusion. NVivo software application was used to analysed and synthesize the findings to identify themes and support the research aim and objectives. Question Addressed: This research addresses the question of how to promote inclusivity, diversity, and inclusion and reduce the prevalence of stereotypes and prejudice. It explores the need to treat individuals based on their unique circumstances and needs rather than relying on generic assumptions. Encourage individuals to adopt a more inclusive approach. Provide managers with responsibility and training that helps them understand the importance of their roles in shaping the workplace culture. Have an equality, diversity, and inclusion manager from a majority background at the senior level who can speak up for underrepresented groups and flag any issues that need addressing. Conclusion: The research emphasizes the importance of promoting equality, diversity, and inclusion practices to create a fairer society. It highlights the need to challenge stereotypes, treat individuals according to their circumstances and needs, and promote a culture of respect and dignity.

Keywords: equality, fairer society, inclusion, diversity

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Authors: Argyro Maria Skourmalla

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Internationalization and management of linguistic diversity in Higher Education (HE) have gained much attention in research in the last few years. Internationalization policies in HE aims at promoting the dual role of Higher Education Institutions (HEIs), civilization and competitiveness. In the context of the European Union, the European Education Area initiative aims at “inclusive national education and training systems” through networking and exchange between HEIs. However, the use of English as a ‘lingua academica’ in the place of the official, national, and regional/minority languages raises questions regarding linguistic diversity, linguistic rights and concerns that have to do with the scientific weakening of these languages. In fact, the European Civil Society Platform for Multilingualism, in the Declaration for Multilingualism in Higher Education, draws attention to the use of English at the expense of other regional/national languages and the impact of English-only language policy on an epistemological level. The above issues were brought up during semi-structured interviews with lecturing staff coming from three multilingual Universities in Europe. Lecturers shared their experiences and the practices they use to manage linguistic diversity in these three Universities. Findings show that even though different languages are used in teaching across disciplines, English -or ‘Globish’ as mentioned during an interview- is widely used in research. Despite English being accepted as the “lingua academica,” issues regarding loss of identity come up

Keywords: higher education, internationalization, linguistic diversity, teaching, research, English

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Authors: Soyhan Egitim

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This case study aims to investigate leadership practices in diversity management in the liberal arts department of a Japanese university. In 2013, the Japanese Ministry of Education, Sports, Science, and Technology (MEXT) revealed their English education reform plan in response to rapid globalization. Based on the new reform plan, Japanese universities would expand their international faculty in order to promote globalization through an increased number of intercultural communication and content-based language classes in English. The study employed a two-phased qualitative approach to gain a deeper understanding of the management strategies employed in diversity management, and the leadership practices influenced those management strategies. In the first phase, a closed-ended qualitative survey was conducted with ten adjunct faculty members from the liberal arts department. The results indicate that syllabus design, grading scheme, textbook choices, and class management policies are strictly regulated by the tenured Japanese faculty. In the second phase, semi-structured interviews were held with international faculty members to understand their personal experiences. Their responses revealed that top-down management approaches are counter-effective in the department’s efforts to promote diversity and thus, a new organizational culture needs to be nurtured to emphasize inclusion alongside diversity. In this regard, the study proposes collaborative leadership as an inclusive leadership practice to minimize power differences in the hierarchy and increase opportunities for inclusion in the rapidly diversifying workforce.

Keywords: collaborative leadership, diversity, inclusion, international faculty, top-down

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Authors: Chidambaram Thamaraiselvan, Carlos Dosoretz

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This work will explore the influence of low voltage electric field, both alternating (AC) and direct (DC) currents, on biofouling control to highly electrically conductive self-supporting carbon nanotubes (CNT) membranes at conditions which encourage bacterial growth. A mutant strain of Pseudomonas putida S12 was used a model bacterium. The antibiofouling studies were performed with flow-through mode connecting an electric circuit in resistive mode. Major emphasis was placed on AC due to its ability of repulsing and inactivating bacteria. The observations indicate that an AC potential >1500 mV, 1 kHz frequency, 100 Ω external resistance on ground side and pulse wave above the offset (+0.45) almost completely prevented attachment of bacteria (>98.5%) and bacterial inactivation (95.3±2.5%). Findings suggest that at the conditions applied, direct electron transfer might be dominant in a decrease of cell viability. AC resulted more effective than DC, both in terms of biofouling reduction compared to cathodic DC and in terms of cell inactivation compared to anodic DC. This electrically polarized CNT membranes offer a viable antibiofouling strategy to hinder biofouling and simplify membrane care during filtration.

Keywords: bacterial attachment, biofouling control, low electric potential, water treatment

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Authors: Surasak Siripornadulsil, Nutt Poomai, Wilailak Siripornadulsil

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Due to a high ethanol demand, the approach for effective ethanol production is important and has been developed rapidly worldwide. Several agricultural wastes are highly abundant in celluloses and the effective cellulose enzymes do exist widely among microorganisms. Accordingly, the cellulose degradation using microbial cellulose to produce a low-cost substrate for ethanol production has attracted more attention. In this study, the cellulose producing bacterial strain has been isolated from rich straw and identified by 16S rDNA sequence analysis as Acinetobacter sp. KKU44. This strain is able to grow and exhibit the cellulose activity. The optimal temperature for its growth and cellulose production is 37 °C. The optimal temperature of bacterial cellulose activity is 60 °C. The cellulose enzyme from Acinetobacter sp. KKU44 is heat-tolerant enzyme. The bacterial culture of 36 h. showed highest cellulose activity at 120 U/mL when grown in LB medium containing 2% (w/v). The capability of Acinetobacter sp. KKU44 to grow in cellulosic agricultural wastes as a sole carbon source and exhibiting the high cellulose activity at high temperature suggested that this strain could be potentially developed further as a cellulose degrading strain for a production of low-cost substrate used in ethanol production.

Keywords: cellulose enzyme, bagasse, rice straw, rice husk, acinetobacter sp. KKU44

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Authors: Adjebli Ahmed, Messis Abdelaziz, Ayeche Riad, Tighilet Karim, Talbi Melissa, Smaili Yanis, Lehri Mokrane, Louardiane Mustapha

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Colorectal cancer is one of the most common types of cancer worldwide, and its incidence has been increasing in recent years. Data and fecal samples from colorectal cancer patients were collected at the Amizour Public Hospital's oncology department (Bejaia, Algeria). Microbiological and cohort study were conducted at the Biological Engineering of Cancers laboratory at the Faculty of Medicine of the University of Bejaia. All the data showed that patients aged between 50 and 70 years were the most affected by colorectal cancer, while the age categories of [30-40] and [40-50] were the least affected. Males were more likely to be at risk of contracting colorectal cancer than females. The most common types of colorectal cancer among the studied population were sigmoid cancer, rectal cancer, transverse colon cancer, and ascending colon cancer. The hereditary factor was found to be more dominant than other risk factors. Bacterial identification revealed the presence of certain pathogenic and opportunistic bacterial genera, such as E. coli, K. pneumoniae, Shigella sp, and Streptococcus group D. These results led us to conclude that dysbiosis of the intestinal microbiome is strongly present in colorectal cancer patients at the EPH of Amizour.

Keywords: microbiome, colorectal cancer, risk factors, bacterial identification

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Authors: Recep Keşli, Cengiz Demir, Onur Türkyılmaz

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Objective: The aim of this study was to determine the prevalence of Mycoplasma hominis and Ureaplasma urealyticum as the causative agents in men with non-gonococcal urethtritis, and anti-bacterial resistance rates. Methods: The Study was carried out in the two Medical Microbiology Laboratories belonging to: Konya Education and Research Hospital and ANS Practice and Research Hospital, Afyon Kocatepe University, between January 2012 and December 2015. Urethral samples were obtained from patients by using a swab. Mycoplasma hominis and Ureaplasma urealyticum were detected by using Mycoplasma IST-2 kit (bio-Mérieux, Marcy l'Étoile, France). Neisseria gonorrhoea was excluded by Gram staining and culture methods. Results: Of all the one hundred and eighty-eight male patients with urethritis, forty M. hominis and forty two U. urealyticum were detected. Resistance rates of M. hominis strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 5 %, 65 %, 25 %, 5 %, 80 %, 20 %, 20 %, 20 %, 5 %, respectively. Resistance rates of U. urealyticum strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 4.7 %, 66.6 %, 23.8 %, 4.75 %, 81 %, 19 %, 19 %, 4.7 % respectively. No resistance was detected against to josamycin, for both the strains. Conclusions: It was concluded that; ciprofloxacin and ofloxacin had the weakest; josamycin, doxycycline, and tetracycline had the strongest in vitro anti-bacterial activity, for treatment of the NGU. So josamycin, doxycycline, and tetracycline should be preferred as the first choice of anti-bacterial agents, for treatment of the patients with non-gonococcal male urethritis.

Keywords: antimicrobial resistance, Mycoplasma hominis, non-gonococcal urethritis, Ureaplasma urealyticum

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

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Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosae (PS1),andPseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazenon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 1012CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Keywords: Acinetobacter baumannii, biodegradation, malathion, organophosphate pesticides

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Authors: A. Hamieh, Z. Olama, H. Holail

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Drinking Water Distribution Systems provide opportunities for microorganisms that enter the drinking water to develop into biofilms. Antimicrobial agents, mainly chlorine, are used to disinfect drinking water, however, there are not yet standardized disinfection strategies with reliable efficacy and development of novel anti-biofilm strategies is still of major concern. In the present study the ability of Lactobacillus acidophilus and Streptomyces sp. cell free supernatants to inhibit the bacterial biofilm formation in Drinking Water Distribution System in Lebanon was investigated. Treatment with cell free supernatants of Lactobacillus acidophilus and Streptomyces sp. at 20% concentration resulted in average biofilm inhibition (52.89 and 39.66% respectively). A preliminary investigation about the mode of action of biofilm inhibition revealed that cell free supernatants showed no bacteriostatic or bactericidal activity against all the tested isolates. Pre-coating wells with supernatants revealed that Lactobacillus acidophilus cell free supernatant inhibited average biofilm formation (62.53%) by altering the adhesion of bacterial isolates to the surface, preventing the initial attachment step, which is important for biofilm production.

Keywords: biofilm, cell free supernatant, distribution system, drinking water, lactobacillus acidophilus, streptomyces sp, adhesion

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Authors: Caterina Calicchio, Talia Sbardella

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The ability to move through geographical and symbolic spaces is key for building new nodes and social relationships. Especially in the framework of language learning, accepting and valuing diversity can help to create a constructive atmosphere of cooperation, innovation, and creativity. Thus, it is important to outline the stages of forming a learning community, focusing on the characteristics that can favor its development. It is known that elements like curiosity and motivation are significant for individual language learning; hence, the study attempts to investigate how factors like openness to diversity and cultural immersion could improve Italian learning and teaching. This paper aims to indicate the factors that could be significant for the development of a Learning Community by presenting a case study on a course on Italian as a second language for beginners: first, the theoretical matrices underlying social learning will be outlined. Secondly, a quantitative study will be described based on an adaptation of the openness to diversity and some insights psychometric scale questionnaire developed at the Umbra Institute. The questionnaire was delivered to 52 American college students with open-ended and closed-ended questions. Students were asked to specify their level of agreement to a set of statements on a six-point Likert scale ranging from (1) Strongly disagree to (6) Strongly agree. The data has been analyzed with a quantitative and qualitative method and has been represented in a pie chart and in a histogram. Moreover, mean and frequency have been calculated. The research findings demonstrate that openness to diversity and challenge enhances cross-cutting skills such as intercultural and communicative competence: through cultural immersion and the facility of speaking with locals, the participants have been able to develop their own Italian L2 language community. The goal is to share with the scientific community some insights to trace possible future lines of research.

Keywords: Italian as second language, language learning, learning community, openness to diversity

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Authors: Musadiq Idris, Sajjad Ali, Syed A. Khaliq, Umer Farooq

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Postmortem record of 217 captive ungulates including Black-buck (n=31), Chinkara (n=20), Hog deer (n=116), Spotted deer (n=35), Red Deer n=(04), and Rusa deer (n=11) submitted to the Veterinary Research Institute, Lahore, Pakistan was analyzed to determine the primary cause of mortality in these animals. The submissions included temporal distribution from Government wildlife captive farms, zoo, and private ownerships, over a three year period (2007-2009). The most common cause of death was found to be trauma (20.27%), followed by parasitic diseases (15.67%), bacterial diseases (11.98%), stillbirths (9.21%), snakebites (2.76%), gut affections (2.30%), neoplasia (1.38%) and starvation (0.92%). The exact cause of death could not be determined in 77 of 217 animals. Pneumonia (8.29%) and tuberculosis (3.69%) were the most common bacterial diseases. Analyses for parasitic infestation revealed tapeworms to be highest (11.05%), followed by roundworms (8.29%) and hemoparasitism (5.07%) (babesiosis and theileriosis). The mortality rate in young ungulates was lower as compared to adults (32.26% and 67.74%). Gender wise data presented higher mortality in females (55.30%) compared to males (44.70%). In conclusion, highest mortality factor in captive ungulates was trauma, followed by parasitic and bacterial infestations/infections of tapeworms and pneumonia, respectively. Furthermore, necropsies provided substantial information on etiology of death and other related epidemiological aspects.

Keywords: age, epidemiology, gender, mortality, ungulates

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Authors: Muhammad Awais Afzal, Lorena Tuchscherr De Hauschopp, Christian Hübner

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Introduction: Autophagy is an evolutionarily conserved lysosome-dependent degradation pathway, which can be induced by extrinsic and intrinsic stressors in living systems to adapt to fluctuating environmental conditions. In the context of inflammatory stress, autophagy contributes to the elimination of invading pathogens, the regulation of innate and adaptive immune mechanisms, and regulation of inflammasome activity as well as tissue damage repair. Lysosomes can be recycled from autolysosomes by the process of autophagic lysosome reformation (ALR), which depends on the presence of several proteins including Spatacsin. Thus ALR contributes to the replenishment of lysosomes that are available for fusion with autophagosomes in situations of increased autophagic turnover, e.g., during bacterial infections, inflammatory stress or sepsis. Objectives: We aimed to assess whether ALR plays a role for cell survival in an in-vitro bacterial infection model. Methods: Mouse embryonic fibroblasts (MEFs) were isolated from wild-type mice and Spatacsin (Spg11-/-) knockout mice. Wild-type MEFs and Spg11-/- MEFs were infected with Staphylococcus aureus (multiplication of infection (MOI) used was 10). After 8 and 16 hours of infection, cell viability was assessed on BD flow cytometer through propidium iodide intake. Bacterial intake by cells was also calculated by plating cell lysates on blood agar plates. Results: in-vitro infection of MEFs with Staphylococcus aureus showed a marked decrease of cell viability in ALR deficient Spatacsin knockout (Spg11-/-) MEFs after 16 hours of infection as compared to wild-type MEFs (n=3 independent experiments; p < 0.0001) although no difference was observed for bacterial intake by both genotypes. Conclusion: Suggesting that ALR is important for the defense of invading pathogens e.g. S. aureus, we observed a marked increase of cell death in an in-vitro infection model in cells with compromised ALR.

Keywords: autophagy, autophagic lysosome reformation, bacterial infections, Staphylococcus aureus

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