Search results for: native microorganisms

Authors: Ashima Sharma

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Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

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Authors: Ragini Verma, J. Ponmozhi

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The oral cavity is home to a wide variety of microorganisms that lead to various diseases and even oral cancer. Despite advancements in the diagnosis and detection at the initial phase, the situation hasn’t improved much. Saliva-on-a-chip is an innovative point-of-care platform for early diagnosis of oral cancer and other oral diseases in live and dead cells using a microfluidic device with a current perspective. Some of the major challenges, like real-time imaging of the oral cancer microbes, high throughput values, obtaining a high spatiotemporal resolution, etc. were faced by the scientific community. Integrated microfluidics and microscopy provide powerful approaches to studying the dynamics of oral pathology, microbe interaction, and the oral microenvironment. Here we have developed a saliva-on-chip (salivary microbes) device to monitor the effect on oral cancer. Adhesion of cancer-causing F. nucleatum; subsp. Nucleatum and Prevotella intermedia in the device was observed. We also observed a significant reduction in the oral cancer growth rate when mortality and morbidity were induced. These results show that this approach has the potential to transform the oral cancer and early diagnosis study.

Keywords: microfluidic device, oral cancer microbes, early diagnosis, saliva-on-chip

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Authors: Faranak Beigmohammadi, Seyed Hadi Peighambardoust, Seyed Jamaledin Peighambardoust

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This study undertakes to investigate the ability of different kinds of nanocomposite films made of cloisite-30B with different percentages of silver and copper oxide nanoparticles incorporated into a low-density polyethylene (LDPE) polymeric matrix by a melt mixing method in order to inhibit the growth of microorganism in soy burger. The number of surviving cell of the total count was decreased by 3.61 log and mold and yeast diminished by 2.01 log after 8 weeks storage at 18 ± 0.5°C below zero, whilst pure LDPE did not has any antimicrobial effect. A composition of 1.3 % cloisite 30B-Ag and 2.7 % cloisite 30B-CuO for total count and 0 % cloisite 30B-Ag and 4 % cloisite 30B-CuO for yeast & mold gave optimum points in combined design test in Design Expert 7.1.5. Suitable microbial models were suggested for retarding above microorganisms growth in soy burger. To validation of optimum point, the difference between the optimum point of nanocomposite film and its repeat was not significant (p<0.05) by one-way ANOVA analysis using SPSS 17.0 software, while the difference was significant for pure film. Migration of metallic nanoparticles into a food stimulant was within the accepted safe level.

Keywords: modeling, nanocomposite film, packaging, soy burger

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Authors: Sanjay Bari, Vinod Ugale, Kamalkishor Patil

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Over the past several years the emergence of micro-organisms resistant to nearly all the class of antimicrobial agents has become a serious public health concern. In the present research, we report the synthesis and in-vitro antimicrobial activity of a new series of novel quinazolino-thiadiazoles 3 (a-j). The synthesized compounds were confirmed by melting point, IR, 1H-NMR, 13C NMR and Mass spectroscopy. In general, the results of the in-vitro antibacterial activity are encouraging, as out of 10 compounds tested, Compound 3f and 3i with a 4-chloro phenyl and 4-nitro phenyl at C-2 of thiadiazolyl of quinazolino-thiadiazoles, displayed the excellent antibacterial and antifungal activities against all the tested microorganisms (Bacterial and Fungal strain) with MIC values of 62.5 μg/mL. It is worth to mention that the combination of two biologically active moieties quinazoline and thiadiazole profoundly influences the biological activity. While evaluating the antimicrobial activity, it was observed that compounds having electron withdrawing groups on thiazole has shown profound activity in comparison to compounds having electron releasing groups. As a result of this study, it can be concluded that halogen substituent on thiazole ring increases antimicrobial activity. Possible improvements in the antimicrobial activity can be further achieved by slight modifications in the substituent’s and/or additional structural activity investigations to have good antimicrobial activity.

Keywords: antifungal, antimicrobial, quinazolino-thiazoles, synthesis

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Authors: T. Bettaieb, S. Soufi, S. Arbaoui

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Stevia rebaudiana Bert. is a natural sweet plant. The leaves contain diterpene glycosides stevioside, rebaudiosides A-F, steviolbioside and dulcoside, which are responsible for its sweet taste and have commercial value all over the world as sugar substitute in foods and medicines. Stevia rebaudiana Bert. is sensitive temperature lower than 9°C. The possibility of its outdoor culture in Tunisian conditions demand genotypes tolerant to low temperatures. In order to evaluate the low temperature tolerance of eight genotypes of Stevia rebaudiana, the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalases (CAT) were measured. Before carrying out the analyses, three genotypes of Stevia were exposed for 1 month at a temperature regime of 18°C during the day and 7°C at night similar to winter conditions in Tunisia. In response to the stress generated by low temperature, antioxidant enzymes activity revealed on native gel and quantified by spectrophotometry showed variable levels according to their degree of tolerance to low temperatures.

Keywords: chilling tolerance, enzymatic activity, stevia rebaudiana bert, low thermal stress

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Authors: Rawichar Chaipojjana, Suttipong Phosuksirikul, Arunsri Leejeerajumnean

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The objective of the study was to select the survival of probiotic strains when exposed to acidic and bile salts condition. Four probiotic strains (Lactobacillus casei subsp. rhamnosus TISTR 047, Lactobacillus casei TISTR 1500, Lactobacillus acidophilus TISTR 1338 and Lactobacillus plantarum TISTR 1465) were cultured in MRS broth and incubated at 35ºC for 15 hours before being inoculated into acidic condition (5 M HCl, pH 2) for 2 hours and bile salt (0.3%, pH 5.8) for 8 hour. The survived probiotics were counted in MRS agar. Among four stains, Lactobacillus casei subsp. rhamnosus TISTR 047 was the highest tolerance specie. Lactobacillus casei subsp. rhamnosus TISTR 047 reduced 6.74±0.07 log CFU/ml after growing in acid and 5.52±0.05 log CFU/ml after growing in bile salt. Then, double emulsion of microorganisms was chosen to encapsulate before spray drying. Spray drying was done with the inlet temperature 170ºC and outlet temperature 80ºC. The results showed that the survival of encapsulated Lactobacillus casei subsp. rhamnosus TISTR 047 after spray drying decreased from 9.63 ± 0.32 to 8.31 ± 0.11 log CFU/ml comparing with non-encapsulated, 9.63 ± 0.32 to 4.06 ± 0.08 log CFU/ml. Therefore, Lactobacillus casei subsp. rhamnosus TISTR 047 would be able to survive in gastrointestinal and spray drying condition.

Keywords: probiotic, acid, bile salt, spray drying

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Authors: Ayman El Behiry, Rasha Nabil Zahran, Reda Tarabees, Eman Marzouk, Musaad Al-Dubaib

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

Keywords: identification, mastitis pathogens, mass spectral, phenotypical

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Authors: Tan Yong Sheng Edgar, Yeong Wai Yee

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Age-related macular degeneration (AMD) is one of the leading cause of blindness. It can cause severe visual loss due to damaged retinal pigment epithelium (RPE). RPE is an important component of the retinal tissue. It functions as a transducing boundary for visual perception making it an essential factor for sight. The RPE also functions as a metabolically complex and functional cell layer that is responsible for the local homeostasis and maintenance of the extra photoreceptor environment. Thus one of the suggested method of treating such diseases would be regenerating these RPE cells. As such, we intend to grow these cells using a synthetic scaffold to provide a stable environment that reduces the batch effects found in natural scaffolds. Stiffness of the scaffold will also be investigated to determine the optimal Young’s modulus for cultivating these cells. The cells will be generated into a monolayer cell sheet and their functions such as formation of tight junctions and gene expression patterns will be assessed to evaluate the cell sheet quality compared to a native RPE tissue.

Keywords: RPE, scaffold, characterization, biomaterials, colloids and nanomedicine

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Authors: Andrés Calvillo Téllez, Marianne Martínez Zanzarric, José Cruz Núñez Pérez

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A modular system that performs basic monitoring of both water and air quality is presented. Monitoring is essential for environmental, aquaculture, and agricultural disciplines, where this type of instrumentation is necessary for data collection. The system uses low-cost components, which allows readings close to those with high-cost probes. The probe collects readings such as the coordinates of the geographical position, as well as the time it records the target parameters of the monitored. The modules or subsystems that make up the probe are the global positioning (GPS), which shows the altitude, latitude, and longitude data of the point where the reading will be recorded, a real-time clock stage, the date marking the time, the module SD memory continuously stores data, data acquisition system, central processing unit, and energy. The system acquires parameters to measure water quality, conductivity, pressure, and temperature, and for air, three types of ammonia, dioxide, and carbon monoxide gases were censored. The information obtained allowed us to identify the schedule of modification of the parameters and the identification of the ideal conditions for the growth of microorganisms in the water.

Keywords: calibration, conductivity, datalogger, monitoring, real time clock, water quality

Procedia PDF Downloads 91

Authors: K. Karapetyan, F. Tkhruni, A. Aghajanyan, T. S. Balabekyan, L. Arstamyan

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Introduction: Globalization of the food supply has created the conditions favorable for the emergence and spread of food-borne and especially dangerous pathogens (EDP) in developing countries. The fresh-cut fruit and vegetable industry is searching for alternatives to replace chemical treatments with biopreservative approaches that ensure the safety of the processed foods product. Antimicrobial compounds of lactic acid bacteria (LAB) possess bactericidal or bacteriostatic activity against intestinal pathogens, spoilage organisms and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus and Salmonella. Endemic strains of LAB were isolated. The strains, showing broad spectrum of antimicrobial activity against food spoiling microorganisms, were selected. The genotyping by 16S rRNA sequencing, GS-PCR, RAPD PCR methods showed that they were presented by Lactobacillus rhamnosus109, L.plantarum 65, L.plantarum 66 and Enterococcus faecium 64 species. LAB are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB strains were isolated from different dairy products from rural households from the highland regions of Armenia. Serially diluted samples were spread on MRS (Merck, Germany) and hydrolyzed milk agar (1,2 % w/v). Single colonies from each LAB were individually inoculated in liquid MRS medium and incubated at 37oC for 24 hours. Culture broth with biomass was centrifuged at 10,000 g during 20 min for obtaining of cell free culture broth (CFC). The antimicrobial substances from CFC broth were purified by the combination of adsorption-desorption and ion-exchange chromatography methods. Separation of bacteriocins was performed using a HPLC method on "Avex ODS" C18 column. Mass analysis of peptides recorded on the device API 4000 in the electron ionization mode. The spot-on-lawn method on the test culture plated in the solid medium was applied. The antimicrobial activity is expressed in arbitrary units (AU/ml). Results. Purification of CFC broth of LAB allowed to obtain partially purified antimicrobial preparations which contains bacteriocins with broad spectrum of antimicrobial activity. Investigation of their main biochemical properties shown, that inhibitory activity of preparations is partially reduced after treatment with proteinase K, trypsin, pepsin, suggesting a proteinaceous nature of bacteriocin-like substances containing in CFC broth. Preparations preserved their activity after heat treatment (50-121 oC, 20 min) and were stable in the pH range 3–8. The results of SDS PAAG electrophoresis show that L.plantarum 66 and Ent.faecium 64 strains have one bacteriocin (BCN) with maximal antimicrobial activity with approximate molecular weight 2.0-3.0 kDa. From L.rhamnosus 109 two BCNs were obtained. Mass spectral analysis indicates that these bacteriocins have peptide bonds and molecular weight of BCN 1 and BCN 2 are approximately 1.5 kDa and 700 Da. Discussion: Thus, our experimental data shown, that isolated endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms of different taxonomic group, such as Salmonella sp., Esherichia coli, Bacillus sp., L.monocytogenes, Proteus mirabilis, Staph. aureus, Ps. aeruginosa. Obtained results proved the perspectives for use of endemic strains in the preservation of foodstuffs. Acknowledgments: This work was realized with financial support of the Project Global Initiatives for Preliferation Prevention (GIPP) T2- 298, ISTC A-1866.

Keywords: antimicrobial activity, bacteriocins, endemic strains, food safety

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Authors: Reginawanti Hindersah, Widiya Septiani Perdanawati, Dewi Azizah Sulaksana, Hidiyah Ayu Ma’rufah

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Maize in some region in East Nusa Tenggara Indonesia bordering Republic Democratic of Timor Leste is important local food crop and commonly cultivated using conventional method without appropriate plant nutrition system so that productivity is still low. A way to enhance local corn yield is adding biofertilizer containing nitrogen (N2) fixing bacteria such as Azotobacter. The purpose of this research was to determine N2 fixation, cytokinin as well as exopolysachharide production capacity of six indigenous Azotobacter strains in pure culture. The N2 fixation capacities of native 3 day old Azotobacter strains added to Ashby Media varied from 0.01 to 0.39 µM/g/hour. Cytokinin production of these strain in liquid culture of N-free Media was 0.11 to 40.04 ppm while exopolysachharide content in liquid culture of Vermani Media varied from 0.4 to 27.3 g/L. This results demonstrate that some local Azotobacter strains might be used as biofertilizer.

Keywords: azotobacter, local isolate, N fixation, phythohormone, exopolysaccaride

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Authors: Kai-Chee Loh, Jingxin Zhang, Yen-Wah Tong

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Hydrolysis and acidogenesis are the rate-controlling steps in an anaerobic digestion (AD) process. Considering that the optimum conditions for each stage can be diverse diverse, the development of a multi-stage AD system is likely to the AD efficiency through individual optimization. In this research, we developed a highly integrate three-stage anaerobic digester (HM3) to combine the advantages of dry AD and wet AD for anaerobic co-digestion of food waste and horse manure. The digester design comprised mainly of three chambers - high-solids hydrolysis, high-solids acidogenesis and wet methanogensis. Through comparing the treatment performance with other two control digesters, HM3 presented 11.2 ~22.7% higher methane yield. The improved methane yield was mainly attributed to the functionalized partitioning in the integrated digester, which significantly accelerated the solubilization of solid organic matters and the formation of organic acids, as well as ammonia in the high-solids hydrolytic and acidogenic stage respectively. Additionally, HM3 also showed the highest volatile solids reduction rate among the three digesters. Real-time PCR and pyrosequencing analysis indicated that the abundance and biodiversity of microorganisms including bacteria and archaea in HM3 was much higher than that in the control reactors.

Keywords: anaerobic digestion, high-solids, food waste and horse manure, microbial community

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Authors: Mabtho Moreroa-Monyelo, Grace Ijoma, Rosina Nkuna, Tonderayi Matambo

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A moving bed biofilm reactor (MBBR) was used for the bioremediation of high strength chemical oxygen demand (COD) Fisher-Tropsch (FT) wastewater. The aerobic MBBR system was operated over 60 days. For metagenomics profile assessment of the targeted 16S sequence of bacteria involved in the bioremediation of the chemical compounds, sludge samples were collected every second day of operation. Parameters such as pH and COD were measured daily to compare the system efficiency as the changedin microbial diversity progressed. The study revealed that pH was a contributing factor to microbial diversity, which further affected the efficiency of the MBBR system. The highest COD removal rate of 86.4% was achieved at pH 8.3. It was observed that when there was more, A higher bacterial diversity led to an improvement in the reduction of COD. Furthermore, an OTUof 4530 was obtained, which were divided into 12 phyla, 27 classes, 44 orders, 74 families, and 138 genera across all sludge samples from the MBBR. A determination of the relative abundance of microorganisms at phyla level indicates that the most abundant phylum on day it was Firmicutes (50%); thereafter, the most abundant phylum changed toProteobacteria.

Keywords: biodegradation, fischer-tropsch wastewater, metagenomics, moving bed biofilm reactor

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Authors: F. Halladj, H. Amellal, S. Benamara

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Current consumer trends go towards natural products defined as the products obtained by a traditional manufacturing method. Vinegar is one of those products marketed; it may be industrially obtained by a submerged (fast) or traditional (slow) processes. The latter exhibited a high quality because of its complex microbiological transformations (or two-stage fermentation) by the native must flora. Moreover, although that Acetic acid bacteria have traditionally been considered to play the leading role in vinegar production, some studies have recently highlighted that also yeasts metabolism can affect traditional vinegar chemical properties in a remarkable way. Thus, the aim of this study was to monitor a traditional slow process of vinegar as applied in the south of Algeria using date with hard texture (Degla-Beida variety) to isolate and identify the involved yeasts in order to select them as starter culture. Phenotypic and molecular analysis show that the non-Saccharomyces were the main yeasts species isolated throughout the alcoholic spontaneous fermentation and they included Hanseniaspora guilliermondii and Torulaspora delbrueckii.

Keywords: date vinegar, traditional production, yeasts, Phenotypic, Algeria

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Authors: Rosa M. Gambier, Jennifer L. Carlson

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In order to revitalize the science curriculum for botany courses for non-science majors, we have introduced the use of the medicinal plants into a first-year botany course. We have connected the use of scientific method, scientific inquiry and active learning in the classroom with the study of Western Traditional Medical Botany. The students have researched models of Botanical medicine and have designed a sustainable medicinal plants garden using native medicinal plants from the northeast. Through the semester, the students have researched their chosen species, planted seeds in the college greenhouse, collected germination ratios, growth ratios and have successfully produced a beginners medicinal plant garden. Phase II of the project will be to tie in SCCCs community outreach goals by involving the public in the expanded development of the garden as a way of sharing learning about medicinal plants and traditional medicine outside the classroom.

Keywords: medicinal plant garden, botany curriculum, active learning, community outreach

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Authors: Margareta Rahayuningsih, Nur Rahayu Utami, Tsabit A. M., Muh. Abdullah

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Mount Ungaran is one area that has remaining natural forest in Central Java, Indonesia. Mount Ungaran consists of several habitats that supporting appropriate areas for flora, fauna, and microorganisms biodiversity, particularly of it is protected by government law and IUCN red list data. Therefore, Mount Ungaran also settled up as AZE (Alliance for Zero Extinction) and IBA (Important Bird Area). The land use for agriculture and plantation reduces forest covered areas. It is serious threat to the existence of biodiversity in Moun Ungaran. This research has been identified community local wisdom that possible to be integrated as ethno-biological research and biodiversity conservation. The result showed at least four local wisdom that possible to be integrated to ethno-biological and biodiversity conservation were Wit Weh Woh (a ceremony of life-giving tree), Grebeg Alas Susuk Wangan (a ceremony for forest protection), Iriban (a ceremony of clean water resource protection), and tingkep tandur (a ceremony for ready-harvested plant protection). It is needed ethno-biological researches of local wisdom-contained values, which essential to be developed as a strategy for biodiversity conservation in Mount Ungaran.

Keywords: Mount Ungaran, local wisdom, biodiversity, fragmentation

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Authors: Abdulrahman Al Lily

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This article spans the domains of education, technology and geo-politics. It uses as an instance the Arab scholarship of education and technology, viewing its scholarly community through the geographical lens of regionalism. It enquires into the power relations among scholars in the Arab region and between scholars in the Arab region and their fellows from outside the region. It addresses the research question: to what extent have region-informed factors affected the scholarly community of education and technology in the Arab region? This question was answered by both qualitative and numerical enquiry, analysing documents, interviews and a survey of native Arabic-speaking scholars. Having analysed the data using the grounded theory approach, two categories of power relations among scholars were identified: power relations within a particular region and power relations across regions. Considering these two categories, a theoretical proposition could be posited that there could be power relationships among scholars that exist on a regional basis. The recommendation is therefore that research should further shed light upon the regionalistic (and thus geographically informed political) dynamics of scholarly communities.

Keywords: education, technology, politics, geography, regionalism, Arab

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Authors: Mamiko Orii

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This study reports on primary school second language teachers’ sources of language insecurity. Furthermore, it aims to develop an in-service training course to reduce anxiety and build sufficient English communication skills. Language/Linguistic insecurity refers to a lack of confidence experienced by language speakers. In particular, second language/non-native learners often experience insecurity, influencing their learning efficacy. While language learner insecurity has been well-documented, research on the insecurity of language teaching professionals is limited. Teachers’ language insecurity or anxiety in target language use may adversely affect language instruction. For example, they may avoid classroom activities requiring intensive language use. Therefore, understanding teachers’ language insecurity and providing continuing education to help teachers to improve their proficiency is vital to improve teaching quality. This study investigated Japanese primary school teachers’ language insecurity. In Japan, teachers are responsible for teaching most subjects, including English, which was recently added as compulsory. Most teachers have never been professionally trained in second language instruction during college teacher certificate preparation, leading to low confidence in English teaching. Primary source of language insecurity is a lack of confidence regarding English communication skills. Their actual use of English in classrooms remains unclear. Teachers’ classroom speech remains a neglected area requiring improvement. A more refined programme for second language teachers could be constructed if we can identify areas of need. Two questionnaires were administered to primary school teachers in Tokyo: (1) Questionnaire A: 396 teachers answered questions (using a 5-point scale) concerning classroom teaching anxiety and general English use and needs for in-service training (Summer 2021); (2) Questionnaire B: 20 teachers answered detailed questions concerning their English use (Autumn 2022). Questionnaire A’s responses showed that over 80% of teachers have significant language insecurity and anxiety, mainly when speaking English in class or teaching independently. Most teachers relied on a team-teaching partner (e.g., ALT) and avoided speaking English. Over 70% of the teachers said they would like to participate in training courses in classroom English. Questionnaire B’s results showed that teachers could use simple classroom English, such as greetings and basic instructions (e.g., stand up, repeat after me), and initiate conversation (e.g., asking questions). In contrast, teachers reported that conversations were mainly carried on in a simple question-answer style. They had difficulty continuing conversations. Responding to learners’ ‘on-the-spot’ utterances was particularly difficult. Instruction in turn-taking patterns suitable in the classroom communication context is needed. Most teachers received grammar-based instruction during their entire English education. They were predominantly exposed to displayed questions and form-focused corrective feedback. Therefore, strategies such as encouraging teachers to ask genuine questions (i.e., referential questions) and responding to students with content feedback are crucial. When learners’ utterances are incorrect or unsatisfactory, teachers should rephrase or extend (recast) them instead of offering explicit corrections. These strategies support a continuous conversational flow. These results offer benefits beyond Japan’s English as a second Language context. They will be valuable in any context where primary school teachers are underprepared but must provide English-language instruction.

Keywords: english as a second/non-native language, in-service training, primary school, teachers’ language insecurity

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Authors: Xia Huo

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The composition and metabolites of the gut microbiota can be altered by environmental pollutants. However, the effect of co-exposure to multiple pollutants on the human gut microbiota has not been sufficiently studied. In this study, gut microorganisms and their metabolites were compared between 33 children from Guiyu and 34 children from Haojiang. The exposure level was assessed by estimating the daily intake (EDI) of polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), 6PPD-quinone (6PPDQ), and metal(loid)s in dust. Significant correlations were found between the EDIs of 6PPDQ, BDE28, PCB52, Ni, Cu, and both the alpha diversity index and specific metabolites in single-element models. The study found that the Bayesian kernel machine regression (BKMR) model showed a negative correlation between the EDIs of five pollutants (6PPDQ, BDE28, PCB52, Ni, and Cu) and the Chao 1 index, particularly beyond the 55th percentile. Furthermore, the EDIs of these five pollutants were positively correlated with the levels of the metabolite 2,4-diaminobutyric acid while negatively correlated with the levels of d-erythro-sphingosine and d-threitol. Our research suggests that exposure to 6PPDQ, BDE28, PCB52, Ni, and Cu in kindergarten dust is associated with alterations in the gut microbiota and its metabolites. These alterations may be associated with neurodevelopmental abnormalities in children.

Keywords: gut microbiota, 6PPDQ, PBDEs, PCBs, metal(loid)s, BKMR

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Authors: Pontsho E. Tseke, Phatu W. Mashela

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Climate-smart agriculture dictates that underusilised indigenous plant, which served as food for local marginalized communities, be assessed for introduction into mainstream agriculture. Most of the underutilised indigenous plants had survived adverse conditions in the wild; with limited information on how the interact with most abiotic and biotic factors. Cucumis myriocarpus leafy vegetable has nutritional, pharmacological and industrial applications, with limited information on how it interacts with effective microorganisms. The objective of this study was to determine the effects vesicular arbuscular mycorrhiza (VAM) on the growth of C. myriocarpus indigenous leafy vegetable under greenhouse conditions. Four-weeks-old seedlings of C. myriocarpus were transplanted into 20-cm-diameter plastic pots. Two weeks after transplanting, VAM was applied at 0, 10, 20, 30, 40, 50, 60 and 70 g Biocult-VAM plant. At 56 days after treatments, plant growth variables of C. myriocarpus with increase Biocult-VAM levels exhibited positive quadratic relations. Plant variables and increasing concentrations of salinity exhibited positive quadric relations, with 95 to 99% associations. Inclusion, Biocult-VAM can be used in sustainable production of C. myriocarpus for functional food security.

Keywords: abiotic, biotic, rhizasphere, sustainable agriculture

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Authors: Michelle Maurer, Antonia Last, Mark S. Gresnigt, Bernhard Hube, Alexander S. Mosig

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The intestinal epithelium forms an essential barrier to prevent translocation of microorganisms, toxins or other potentially harmful molecules into the bloodstream. In particular, dendritic cells of the intestinal epithelium orchestrate an adapted response of immune tolerance to commensals and immune defense against invading pathogens. Systemic inflammation is typically associated with a dysregulation of this adapted immune response and is accompanied by a disruption of the epithelial and endothelial gut barrier which enables dissemination of pathogens within the human body. To understand the pathophysiological mechanisms underlying the inflammation-associated gut barrier breakdown, it is crucial to elucidate the complex interplay of the host and the intestinal microbiome. A microfluidically perfused three-dimensional intestine-on-chip model was established to emulate these processes in the presence of immune cells, commensal bacteria, and facultative pathogens. Multi-organ tissue flow (MOTiF) biochips made from polystyrene were used for microfluidic perfusion of the intestinal tissue model. The biochips are composed of two chambers separated by a microporous membrane. Each chamber is connected to inlet and outlet channels allowing independent perfusion of the individual channels and application of microfluidic shear stress. Human umbilical vein endothelial cells (HUVECs), monocyte-derived macrophages and intestinal epithelial cells (Caco-2) were assembled on the biochip membrane. Following 7 – 14 days of growth in the presence of physiological flow conditions, the epithelium was colonized with the commensal bacterium Lactobacillus rhamnosus, while the endothelium was perfused with peripheral blood mononuclear cells (PBMCs). Additionally, L. rhamnosus was co-cultivated with the opportunistic fungal pathogen Candida albicans. Within one week of perfusion, the epithelial cells formed self-organized and well-polarized villus- and crypt-like structures that resemble essential morphological characteristics of the human intestine. Dendritic cells were differentiated in the epithelial tissue that specifically responds to bacterial lipopolysaccharide (LPS) challenge. LPS is well-tolerated at the luminal epithelial side of the intestinal model without signs of tissue damage or induction of an inflammatory response, even in the presence of circulating PBMC at the endothelial lining. In contrast, LPS stimulation at the endothelial side of the intestinal model triggered the release of pro-inflammatory cytokines such as TNF, IL-1β, IL-6, and IL-8 via activation of macrophages residing in the endothelium. Perfusion of the endothelium with PBMCs led to an enhanced cytokine release. L. rhamnosus colonization of the model was tolerated in the immune competent tissue model and was demonstrated to reduce damage induced by C. albicans infection. A microfluidic intestine-on-chip model was developed to mimic a systemic infection with a dysregulated immune response under physiological conditions. The model facilitates the colonization of commensal bacteria and co-cultivation with facultative pathogenic microorganisms. Both, commensal bacteria alone and facultative pathogens controlled by commensals, are tolerated by the host and contribute to cell signaling. The human intestine-on-chip model represents a promising tool to mimic microphysiological conditions of the human intestine and paves the way for more detailed in vitro studies of host-microbiota interactions under physiologically relevant conditions.

Keywords: host-microbiota interaction, immune tolerance, microfluidics, organ-on-chip

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Authors: Alicja Zapolnik-Plachetka

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The paper examines the impact of the Members’ of the European Parliament (MEPs) language choice on the linguistic quality of their political discourse as delivered by the interpreters. The study, designed by the author, who is an EU interpreter herself, consisted of three phases. First, a number of speeches of Polish and Spanish MEPs were analyzed to determine whether the incidence of use of certain figures of speech depending on whether the speech had been delivered in English or their respective mother tongue. Then the use of figures of speech was also analyzed based on speeches by some British MEPs, in order to determine what was the incidence for the native users of English. Subsequently, the speeches were compared with their interpretations to find out whether the interpreters managed to convey accurately the means of oratory used by the MEPs. The final result shows that in case of institutional environments dependant on simultaneous interpretation the speakers’ choices can, in fact, influence the linguistic quality of the political communication.

Keywords: content accuracy, European Parliament, political discourse, simultaneous interpreting

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Authors: Rupa Rani, Vipin Kumar

Abstract:

Endosulfan, an organochlorine pesticide is of environmental concern due to its apparent persistence and toxicity. It has been reported as contaminants in soil, air, and water and is bioaccumulated and magnified in ecosystems. The combined use of microorganisms and plants has great potential for remediating soil contaminated with organic compounds such as pesticides. The objective of this study was to evaluate whether the bacterial inoculation influences plant growth promotion, endosulfan degradation in soil and endosulfan accumulation in different plant parts. Lycopersicon esculentum L. (Tomato) was grown in endosulfan spiked soil and inoculated with endosulfan tolerant bacterial strains. Endosulfan residues from different parts of plants and soil were extracted and estimated by using gas chromatograph equipped with 63Ni electron capture detector (GC-ECD). The inoculation of bacterial strains into the soil with plants showed a beneficial effect on endosulfan degradation and plant biomass production. Maximum endosulfan (90%) degradation was observed after 120 days of bacterial inoculation in the soil. Furthermore, there was significantly less endosulfan accumulation in roots and shoots of bacterial strains inoculated plants as compared to uninoculated plants. The results show the effectiveness of inoculated endosulfan tolerant bacterial strains to increase the remediation of endosulfan contaminated soil.

Keywords: organochlorine pesticides, endosulfan, degradation, plant-bacteria partnerships

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Authors: G. Surendra Babu

Abstract:

We all know the importance of mineral wealth, which has been buried inside the layers of the earth for decades. These are the natural energy sources that are used in our day to day life like fuel, electricity, construction, etc. but the process of extraction causes damage to the nature that can’t be returned back and which are left over after completion of mining we can see these are barren from decades these remain unused degraded land. Most of them are covered with vegetation before the start during mining which damages the native vegetation of the region and disturbs the watershed boundary of the regions and it also disturbs the biodiversity of the reign. The major motto of the study is to understand the various issues that are found and to understand various methods of reclamations process that are suitable for revegetating and also variously practiced which are carried out in the different case studies and government guidelines procedure of lease licenses which includes the environmental clearances and also to study the vegetation pattern according to the major issues identified. And finally suggesting the new guidelines with respect to the old guidelines which helps in the revegetation of the mine-sites which helps in establishing of its own sustainable ecosystem in future.

Keywords: reclamation, open-pit mining, revegetation, reclamation methods

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Authors: David S. Rosenstein

Abstract:

The author believes that modern spoken languages should be equally difficult (or easy) to learn, since all normal children learning their native languages do so at approximately the same rate and with the same competence, progressing from easy to more complex grammar and syntax in the same way. Why then, do some languages seem more difficult than others? Perhaps people are referring to the written language, where it may be true that mastering Chinese requires more time than French, which in turn requires more time than Spanish. But this may be marginal, since Chinese and French children quickly catch up to their Spanish peers in reading comprehension. Rather, the real differences in difficulty derive from two sources: hardened L1 language habits trying to cope with contrasting L2 habits; and unfamiliarity with unique L2 characteristics causing faulty expectations. It would seem that effective L2 teaching and learning must take these two sources of difficulty into consideration. The author feels that the latter (faulty expectations) causes the greatest difficulty, making effective teaching and learning somewhat different for each given foreign language. Examples from Chinese and other languages are presented.

Keywords: learning different languages, language learning difficulties, faulty language expectations

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Authors: Asad Ali, Asif Jamal

Abstract:

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 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: bacillus species, biocontrol agent, biopesticides, phytopathogens

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Authors: Daniel Olshansky, Ramiro Rodrıguez Colmeiro

Abstract:

Relay Mining presents a scalable solution employing probabilistic mechanisms and crypto-economic incentives to estimate RPC volume usage, facilitating decentralized multitenant rate limiting. Network traffic from individual applications can be concurrently serviced by multiple RPC service providers, with costs, rewards, and rate limiting governed by a native cryptocurrency on a distributed ledger. Building upon established research in token bucket algorithms and distributed rate-limiting penalty models, our approach harnesses a feedback loop control mechanism to adjust the difficulty of mining relay rewards, dynamically scaling with network usage growth. By leveraging crypto-economic incentives, we reduce coordination overhead costs and introduce a mechanism for providing RPC services that are both geopolitically and geographically distributed.

Keywords: remote procedure call, crypto-economic, commit-reveal, decentralization, scalability, blockchain, rate limiting, token bucket

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Authors: U. Umadevi, T. Umakanthan

Abstract:

Globally, mycotic dermatitis is very common but there is no single proven specific allopathic treatment regimen. In this study, domestic animals with skin diseases of different age and breed from geographically varied regions of Tamil Nadu state, India were employed. Most of them have had previous treatment with native and allopathic medicines without success. Clinically, the skin lesions were found to be mild to severe. The trial animals were treated with poly herbal formulation (ointment) prepared using the indigenous medicinal plants – viz Andrographis paniculata, Lawsonia inermis and Madhuca longifolia. Allopathic antifungal drugs and ointments, povidone iodine and curabless (Terbinafine HCl, Ofloxacin, Ornidazole, Clobetasol propionate) were used in control. Comparatively, trial animals were found to have lesser course of treatment time and higher recovery rate than control. In Ethnoveterinary, this combination was tried for the first time. This herbal formulation is economical and an alternative for skin diseases.

Keywords: allopathic drugs, dermatitis, domestic animals, poly herbal formulation

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Authors: Ruta Mickiene, Audrius Maruska, Ona Ragazinskiene

Abstract:

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

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Authors: Soheil Behdarvandirad

Abstract:

The relationship between depression and self-focused language has been well documented. The more depressed a person is, the more "I"s, "me"s, and "my"s they will use. The present study attempted to factor in the impact of mood and examine whether negative mood has self-focused impacts similar to those of depression. For this purpose, 160 Iranian native speakers of Farsi were divided into three experimental groups of negative, neutral, and positive groups. After completing the BDI-II inventory and depression measurement, they were presented with pretested mood stimuli (3 separate videos to induce the target moods). Finally, they were asked to write between 10 to 20 minutes about a topic that asked them to freely write about their state of life, how you feel about it and the reasons that had shaped their current life circumstances. While the significant correlation between depression and I-talk was observed, negative mood led to more we-talk in general and seemed to even push the participants away from self-rumination. It seems that it is an emotion-regulatory strategy that participants subconsciously adopt to distract themselves from the disturbing mood. However, negative mood intensified the self-focused language among depressed participants. Such results can be further studied by examining brain areas that are more involved in self-perception and particularly in precuneus.

Keywords: self-focused language, depression, mood, precuneus

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