Search results for: bacterial aggregation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1445

Search results for: bacterial aggregation

335 Medial Temporal Tau Predicts Memory Decline in Cognitively Unimpaired Elderly

Authors: Angela T. H. Kwan, Saman Arfaie, Joseph Therriault, Zahra Azizi, Firoza Z. Lussier, Cecile Tissot, Mira Chamoun, Gleb Bezgin, Stijn Servaes, Jenna Stevenon, Nesrine Rahmouni, Vanessa Pallen, Serge Gauthier, Pedro Rosa-Neto

Abstract:

Alzheimer’s disease (AD) can be detected in living people using in vivo biomarkers of amyloid-β (Aβ) and tau, even in the absence of cognitive impairment during the preclinical phase. [¹⁸F]-MK-6420 is a high affinity positron emission tomography (PET) tracer that quantifies tau neurofibrillary tangles, but its ability to predict cognitive changes associated with early AD symptoms, such as memory decline, is unclear. Here, we assess the prognostic accuracy of baseline [18F]-MK-6420 tau PET for predicting longitudinal memory decline in asymptomatic elderly individuals. In a longitudinal observational study, we evaluated a cohort of cognitively normal elderly participants (n = 111) from the Translational Biomarkers in Aging and Dementia (TRIAD) study (data collected between October 2017 and July 2020, with a follow-up period of 12 months). All participants underwent tau PET with [¹⁸F]-MK-6420 and Aβ PET with [¹⁸F]-AZD-4694. The exclusion criteria included the presence of head trauma, stroke, or other neurological disorders. There were 111 eligible participants who were chosen based on the availability of Aβ PET, tau PET, magnetic resonance imaging (MRI), and APOEε4 genotyping. Among these participants, the mean (SD) age was 70.1 (8.6) years; 20 (18%) were tau PET positive, and 71 of 111 (63.9%) were women. A significant association between baseline Braak I-II [¹⁸F]-MK-6240 SUVR positivity and change in composite memory score was observed at the 12-month follow-up, after correcting for age, sex, and years of education (Logical Memory and RAVLT, standardized beta = -0.52 (-0.82-0.21), p < 0.001, for dichotomized tau PET and -1.22 (-1.84-(-0.61)), p < 0.0001, for continuous tau PET). Moderate cognitive decline was observed for A+T+ over the follow-up period, whereas no significant change was observed for A-T+, A+T-, and A-T-, though it should be noted that the A-T+ group was small.Our results indicate that baseline tau neurofibrillary tangle pathology is associated with longitudinal changes in memory function, supporting the use of [¹⁸F]-MK-6420 PET to predict the likelihood of asymptomatic elderly individuals experiencing future memory decline. Overall, [¹⁸F]-MK-6420 PET is a promising tool for predicting memory decline in older adults without cognitive impairment at baseline. This is of critical relevance as the field is shifting towards a biological model of AD defined by the aggregation of pathologic tau. Therefore, early detection of tau pathology using [¹⁸F]-MK-6420 PET provides us with the hope that living patients with AD may be diagnosed during the preclinical phase before it is too late.

Keywords: alzheimer’s disease, braak I-II, in vivo biomarkers, memory, PET, tau

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334 Biogenic Amines Production during RAS Cheese Ripening

Authors: Amr Amer

Abstract:

Cheeses are among those high-protein-containing foodstuffs in which enzymatic and microbial activities cause the formation of biogenic amines from amino acids decarboxylation. The amount of biogenic amines in cheese may act as a useful indicator of the hygienic quality of the product. In other words, their presence in cheese is related to its spoilage and safety. Formation of biogenic amines during Ras cheese (Egyptian hard cheese) ripening was investigated for 4 months. Three batches of Ras cheese were manufactured using Egyptian traditional method. From each batch, Samples were collected at 1, 7, 15, 30, 60, 90 and 120 days after cheese manufacture. The concentrations of biogenic amines (Tyramine, Histamine, Cadaverine and Tryptamine) were analyzed by high performance liquid chromatography (HPLC). There was a significant increased (P<0.05) in Tyramine levels from 4.34± 0.07 mg|100g in the first day of storage till reached 88.77± 0.14 mg|100g at a 120-day of storage. Also, Histamine and Cadaverine levels had the same increased pattern of Tyramine reaching 64.94± 0.10 and 28.28± 0.08 mg|100g in a 120- day of storage, respectively. While, there was a fluctuation in the concentration of Tryptamine level during ripening period as it decreased from 3.24± 0.06 to 2.66± 0.11 mg|100g at 60-day of storage then reached 5.38±0.08 mg|100g in a 120- day of storage. Biogenic amines can be formed in cheese during production and storage: many variables, as pH, salt concentration, bacterial activity as well as moisture, storage temperature and ripening time, play a relevant role in their formation. Comparing the obtained results with the recommended standard by Food and Drug Administration "FDA" (2001), High levels of biogenic amines in various Ras cheeses consumed in Egypt exceeded the permissible value (10 mg%) which seemed to pose a threat to public health. In this study, presence of high concentrations of biogenic amines (Tyramine, Histamine, cadaverine and Tryptamine) in Egyptian Ras cheeses reflects the bad hygienic conditions under which they produced and stored. Accordingly, the levels of biogenic amines in different cheeses should be come in accordance with the safe permissible limit recommended by FDA to ensure human safety.

Keywords: Ras cheese, biogenic amines, tyramine, histamine, cadaverine

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333 Exploring the Correlation between Population Distribution and Urban Heat Island under Urban Data: Taking Shenzhen Urban Heat Island as an Example

Authors: Wang Yang

Abstract:

Shenzhen is a modern city of China's reform and opening-up policy, the development of urban morphology has been established on the administration of the Chinese government. This city`s planning paradigm is primarily affected by the spatial structure and human behavior. The subjective urban agglomeration center is divided into several groups and centers. In comparisons of this effect, the city development law has better to be neglected. With the continuous development of the internet, extensive data technology has been introduced in China. Data mining and data analysis has become important tools in municipal research. Data mining has been utilized to improve data cleaning such as receiving business data, traffic data and population data. Prior to data mining, government data were collected by traditional means, then were analyzed using city-relationship research, delaying the timeliness of urban development, especially for the contemporary city. Data update speed is very fast and based on the Internet. The city's point of interest (POI) in the excavation serves as data source affecting the city design, while satellite remote sensing is used as a reference object, city analysis is conducted in both directions, the administrative paradigm of government is broken and urban research is restored. Therefore, the use of data mining in urban analysis is very important. The satellite remote sensing data of the Shenzhen city in July 2018 were measured by the satellite Modis sensor and can be utilized to perform land surface temperature inversion, and analyze city heat island distribution of Shenzhen. This article acquired and classified the data from Shenzhen by using Data crawler technology. Data of Shenzhen heat island and interest points were simulated and analyzed in the GIS platform to discover the main features of functional equivalent distribution influence. Shenzhen is located in the east-west area of China. The city’s main streets are also determined according to the direction of city development. Therefore, it is determined that the functional area of the city is also distributed in the east-west direction. The urban heat island can express the heat map according to the functional urban area. Regional POI has correspondence. The research result clearly explains that the distribution of the urban heat island and the distribution of urban POIs are one-to-one correspondence. Urban heat island is primarily influenced by the properties of the underlying surface, avoiding the impact of urban climate. Using urban POIs as analysis object, the distribution of municipal POIs and population aggregation are closely connected, so that the distribution of the population corresponded with the distribution of the urban heat island.

Keywords: POI, satellite remote sensing, the population distribution, urban heat island thermal map

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332 Biophysical Characterization of the Inhibition of cGAS-DNA Sensing by KicGAS, Kaposi's Sarcoma-Associated Herpesvirus Inhibitor of cGAS

Authors: D. Bhowmik, Y. Tian, Q. Yin, F. Zhu

Abstract:

Cyclic GMP-AMP synthase (cGAS), recognises cytoplasmic double-stranded DNA (dsDNA), indicative of bacterial and viral infections, as well as the leakage of self DNA by cellular dysfunction and stresses, to elicit the host's immune responses. Viruses also have developed numerous strategies to antagonize the cGAS-STING pathway. Kaposi's sarcoma-associated herpesvirus (KSHV) is a human DNA tumor virus that is the causative agent of Kaposi’s sarcoma and several other malignancies. To persist in the host, consequently causing diseases, KSHV must overcome the host innate immune responses, including the cGAS-STING DNA sensing pathway. We already found that ORF52 or KicGAS (KSHV inhibitor of cGAS), an abundant and basic gamma herpesvirus-conserved tegument protein, directly inhibits cGAS enzymatic activity. To better understand the mechanism, we have performed the biochemical and structural characterization of full-length KicGAS and various mutants in regarding binding to DNA. We observed that KicGAS is capable of self-association and identified the critical residues involved in the oligomerization process. We also characterized the DNA-binding of KicGAS and found that KicGAS cooperatively oligomerizes along the length of the double stranded DNA, the highly conserved basic residues at the c-terminal disordered region are crucial for DNA recognition. Deficiency in oligomerization also affects DNA binding. Thus DNA binding by KicGAS sequesters DNA and prevents it from being detected by cGAS, consequently inhibiting cGAS activation. KicGAS homologues also inhibit cGAS efficiently, suggesting inhibition of cGAS is evolutionarily conserved mechanism among gamma herpesvirus. These results highlight the important viral strategy to evade this innate immune sensor.

Keywords: Kaposi's sarcoma-associated herpesvirus, KSHV, cGAS, DNA binding, inhibition

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331 Treatment of Porphyromonas gingivalis Induced Gingivitis in Albino Rats with Tetracycline-Loaded Nanochitosan, an Immunohistochemical Analysis

Authors: Rania Hanafi Said, Rasha Mohamed Taha

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Background: By using nanoparticles as drug delivery, it may be possible to avoid the drawbacks of systemic antibiotic dosing, including bacterial antibiotic resistance. The goal of this study was to see how well tetracycline loaded on nanochitosan worked to treat gingival inflammation in albino rats caused by Porphyromonas gingivalis. The study analyzed immunohistochemically the localization of the pro-inflammatory cytokine Interleukin-1beta (IL-1β). Material and methods: In this study, fifty mature male albino rats weighing 150 to 180 grams each were used. They were randomly divided into five groups. We checked for weight changes in rats. Ten male albino rats were included in Group I, which served as a negative control group. Ten rats were included in Group II, where they were exposed once to Porphyromonas. Group III contained ten rats, which were treated the same as Group II plus daily injections of diluted tetracycline powder at the infection sites. Ten rats in Group IV received the same procedure as those in Group II before receiving daily injections of nanochitosan at the injection sites. Finally, Group V, which had ten rats. Following the same protocol as Group II, they received localized injections of tetracycline loaded on nanochitosan once daily. Rats' gingivae were extracted and prepared after they were anesthetized. The biopsies were examined histologically and immunohistochemically by light microscopy. Results: Groups I and V had a nearly normal histological appearance of gingival tissue. In Groups II, III, and IV, degeneration was seen because the epithelial cells were bigger, collagen fibers were pulling away from the lamina propria connective tissue, and the basement membranes had come to an end. There was no discernible difference between groups V and I when they were examined immunohistochemically. Conclusion: The use of nano chitosan as a tetracycline carrier is a novel technique to overcome the drug's rising level of resistance.

Keywords: Immunohistochemistry, Nanochitosan, porphyromonas gingivitis, Tetracycline

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330 Characterization of Herberine Hydrochloride Nanoparticles

Authors: Bao-Fang Wen, Meng-Na Dai, Gao-Pei Zhu, Chen-Xi Zhang, Jing Sun, Xun-Bao Yin, Yu-Han Zhao, Hong-Wei Sun, Wei-Fen Zhang

Abstract:

A drug-loaded nanoparticles containing berberine hydrochloride (BH/FA-CTS-NPs) was prepared. The physicochemical characterizations of BH/FA-CTS-NPs and the inhibitory effect on the HeLa cells were investigated. Folic acid-conjugated chitosan (FA-CTS) was prepared by amino reaction of folic acid active ester and chitosan molecules; BH/FA-CTS-NPs were prepared using ionic cross-linking technique with BH as a model drug. The morphology and particle size were determined by Transmission Electron Microscope (TEM). The average diameters and polydispersity index (PDI) were evaluated by Dynamic Light Scattering (DLS). The interaction between various components and the nanocomplex were characterized by Fourier Transform Infrared Spectroscopy (FT-IR). The entrapment efficiency (EE), drug-loading (DL) and in vitro release were studied by UV spectrophotometer. The effect of cell anti-migratory and anti-invasive actions of BH/FA-CTS-NPs were investigated using MTT assays, wound healing assays, Annexin-V-FITC single staining assays, and flow cytometry, respectively. HeLa nude mice subcutaneously transplanted tumor model was established and treated with different drugs to observe the effect of BH/FA-CTS-NPs in vivo on HeLa bearing tumor. The BH/FA-CTS-NPs prepared in this experiment have a regular shape, uniform particle size, and no aggregation phenomenon. The results of DLS showed that mean particle size, PDI and Zeta potential of BH/FA-CTS NPs were (249.2 ± 3.6) nm, 0.129 ± 0.09, 33.6 ± 2.09, respectively, and the average diameter and PDI were stable in 90 days. The results of FT-IR demonstrated that the characteristic peaks of FA-CTS and BH/FA-CTS-NPs confirmed that FA-CTS cross-linked successfully and BH was encapsulated in NPs. The EE and DL amount were (79.3 ± 3.12) % and (7.24 ± 1.41) %, respectively. The results of in vitro release study indicated that the cumulative release of BH/FA-CTS NPs was (89.48±2.81) % in phosphate-buffered saline (PBS, pH 7.4) within 48h; these results by MTT assays and wund healing assays indicated that BH/FA-CTS NPs not only inhibited the proliferation of HeLa cells in a concentration and time-dependent manner but can induce apoptosis as well. The subcutaneous xenograft tumor formation rate of human cervical cancer cell line HeLa in nude mice was 98% after inoculation for 2 weeks. Compared with BH group and BH/CTS-NPs group, the xenograft tumor growth of BH/FA-CTS-NPs group was obviously slower; the result indicated that BH/FA-CTS-NPs could significantly inhibit the growth of HeLa xenograft tumor. BH/FA-CTS NPs with the sustained release effect could be prepared successfully by the ionic crosslinking method. Considering these properties, block proliferation and impairing the migration of the HeLa cell line, BH/FA-CTS NPs could be an important compound for consideration in the treatment of cervical cancer.

Keywords: folic-acid, chitosan, berberine hydrochloride, nanoparticles, cervical cancer

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329 Evaluating the Management of Febrile Infants (Less than 90 Days) Presenting to Tallaght Ed- Completed Audit Cycle

Authors: Amel Osman, Stewart McKenna

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Aim: Fever may present as the sole sign of a serious underlying infection in young infants. Febrile Infants aged less than 90 days are at an elevated susceptibility to invasive bacterial infections, thus presenting a challenge in ensuring the appropriate management of these cases. This study aims to ensure strict adherence to NICE guidelines for the management of fever in infants between 0 and 90 days presenting to Tallaght Hospital ED. A comprehensive audit, followed by a re-audit, was conducted to enhance the quality of care delivered to these patients. In accordance with NICE guidelines, all febrile infants should undergo blood tests. Additionally, LP should be performed in all neonates under 28 days, infants displaying signs of illness, and those with WCC below 5 or above 15. Method: A retrospective case review was performed, encompassing all patients aged between 0 to 90 days who presented with fever at Tallaght ED. Data retrieval was conducted from electronic records on two separate occasions, six months apart. The evaluation encompassed the assessment of body temperature as well as both partial and full septic workups. Results: Over the study period, 150 infants presented to the ED with fever in the initial audit, and 120 in the re-audit. In the first study, 81 patients warranted a full septic workup as per NICE, but only 48 received it. Conversely, 40 patients met criteria for a partial septic workup, with 12 undergoing blood tests. In the second study, 73 patients qualified for a full septic workup, of which 52 were completed. Additionally, 27 patients were indicated for a partial workup, with 20 undergoing blood tests. Conclusion: Managing febrile infants under three months of age presenting to Tallaght ED remains a persistent challenge, underscoring the need for continuous educational initiatives to guarantee that these patients receive the requisite assessments and treatments.

Keywords: infants, fever, septic workup, tallaght

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328 In silico Subtractive Genomics Approach for Identification of Strain-Specific Putative Drug Targets among Hypothetical Proteins of Drug-Resistant Klebsiella pneumoniae Strain 825795-1

Authors: Umairah Natasya Binti Mohd Omeershffudin, Suresh Kumar

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Klebsiella pneumoniae, a Gram-negative enteric bacterium that causes nosocomial and urinary tract infections. Particular concern is the global emergence of multidrug-resistant (MDR) strains of Klebsiella pneumoniae. Characterization of antibiotic resistance determinants at the genomic level plays a critical role in understanding, and potentially controlling, the spread of multidrug-resistant (MDR) pathogens. In this study, drug-resistant Klebsiella pneumoniae strain 825795-1 was investigated with extensive computational approaches aimed at identifying novel drug targets among hypothetical proteins. We have analyzed 1099 hypothetical proteins available in genome. We have used in-silico genome subtraction methodology to design potential and pathogen-specific drug targets against Klebsiella pneumoniae. We employed bioinformatics tools to subtract the strain-specific paralogous and host-specific homologous sequences from the bacterial proteome. The sorted 645 proteins were further refined to identify the essential genes in the pathogenic bacterium using the database of essential genes (DEG). We found 135 unique essential proteins in the target proteome that could be utilized as novel targets to design newer drugs. Further, we identified 49 cytoplasmic protein as potential drug targets through sub-cellular localization prediction. Further, we investigated these proteins in the DrugBank databases, and 11 of the unique essential proteins showed druggability according to the FDA approved drug bank databases with diverse broad-spectrum property. The results of this study will facilitate discovery of new drugs against Klebsiella pneumoniae.

Keywords: pneumonia, drug target, hypothetical protein, subtractive genomics

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327 Phytochemical Screening, Antioxidant and Antibacterial Activity of Annona cherimola Mill

Authors: Arun Jyothi Bheemagani, Chakrapani Pullagummi, Anupalli Roja Rani

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Exploration of the chemical constituents of the plants and pharmacological screening may provide us the basis for the development of novel agents. Plants have provided us some of the very important life saving drugs used in the modern medicine. The aim of our work was to screen the phytochemical constituents and antimicrobial and antioxidant activities of methanol extract of leaves of Annona cherimola Mill plant from Tirumala forest, Tirupathi. It was originally called Chirimuya by the Inca people who lived where it was growing in the Andes of South America, is an edible fruit-bearing species of the genus Annona from the family Annonaceae. Annona cherimola Mill is a multipurpose tree with edible fruits and is one of the sources of the medicinal products. The antibacterial activity was measured by agar well diffusion method; the diameter of the zone of bacterial growth inhibition was measured after incubation of plates. The inhibitory effect was studied against the pathogenic bacteria (Klebsiella pneumonia, Bacillus subtilis, Staphylococcus aureus and Escherichia coli (E. coli). Antioxidant assays were also performed for the same extracts by spectrophotometric methods using known standard antioxidants as reference. The studied plant extracts were found to be very effective against the pathogenic microorganisms tested. The methanolic extract of Annona cherimola Mill from showed maximum activity against Escherichia coli and Staphylococcus aureus and the least concentration required showing the activity was 0.5mg/ml. Phytochemical screening of the plants revealed the presence of flavonoids, alkaloids, steroids and carbohydrates. Good presence of antioxidants was also found in the methanolic extracts.

Keywords: annona cherimola, phytochemicals, antioxidant and antibacterial activity, methanol extract

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326 An Unbiased Profiling of Immune Repertoire via Sequencing and Analyzing T-Cell Receptor Genes

Authors: Yi-Lin Chen, Sheng-Jou Hung, Tsunglin Liu

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Adaptive immune system recognizes a wide range of antigens via expressing a large number of structurally distinct T cell and B cell receptor genes. The distinct receptor genes arise from complex rearrangements called V(D)J recombination, and constitute the immune repertoire. A common method of profiling immune repertoire is via amplifying recombined receptor genes using multiple primers and high-throughput sequencing. This multiplex-PCR approach is efficient; however, the resulting repertoire can be distorted because of primer bias. To eliminate primer bias, 5’ RACE is an alternative amplification approach. However, the application of RACE approach is limited by its low efficiency (i.e., the majority of data are non-regular receptor sequences, e.g., containing intronic segments) and lack of the convenient tool for analysis. We propose a computational tool that can correctly identify non-regular receptor sequences in RACE data via aligning receptor sequences against the whole gene instead of only the exon regions as done in all other tools. Using our tool, the remaining regular data allow for an accurate profiling of immune repertoire. In addition, a RACE approach is improved to yield a higher fraction of regular T-cell receptor sequences. Finally, we quantify the degree of primer bias of a multiplex-PCR approach via comparing it to the RACE approach. The results reveal significant differences in frequency of VJ combination by the two approaches. Together, we provide a new experimental and computation pipeline for an unbiased profiling of immune repertoire. As immune repertoire profiling has many applications, e.g., tracing bacterial and viral infection, detection of T cell lymphoma and minimal residual disease, monitoring cancer immunotherapy, etc., our work should benefit scientists who are interested in the applications.

Keywords: immune repertoire, T-cell receptor, 5' RACE, high-throughput sequencing, sequence alignment

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325 Plant Growth and Yield Enhancement of Soybean by Inoculation with Symbiotic and Nonsymbiotic Bacteria

Authors: Timea I. Hajnal-Jafari, Simonida S. Đurić, Dragana R. Stamenov

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Microbial inoculants from the group of symbiotic-nitrogen-fixing rhizobia are well known and widely used in production of legumes. On the other hand, nonsymbiotic plant growth promoting rhizobacteria (PGPR) are not commonly used in practice. The objective of this study was to examine the effects of soybean inoculation with symbiotic and nonsymbiotic bacteria on plant growth and seed yield of soybean. Microbiological activity in rhizospheric soil was also determined. The experiment was set up using a randomized block system in filed conditions with the following treatments: control-no inoculation; treatment 1-Bradyrhizobium japonicum; treatment 2-Azotobacter sp.; treatment 3-Bacillus sp..In the flowering stage of growth (FS) the number of nodules per plant (NPP), root length (RL), plant height (PH) and weight (PW) were measured. The number of pod per plant (PPP), number of seeds per pod (SPP) and seed weight per plant (SWP) were recorded at the end of vegetation period (EV). Microbiological analyses of soil included the determination of total number of bacteria (TNB), number of fungi (FNG), actinomycetes (ACT) and azotobacters (AZB) as well as the activity of the dehydrogenase enzyme (DHA). The results showed that bacterial inoculation led to the formation of root nodules regardless of the treatments with statistically no significant difference. Strong nodulation was also present in control treatment. RL and PH were positively influenced by inoculation with Azotobacter sp. and Bacillus sp., respectively. Statistical analyses of the number of PPP, SPP, and SWP showed no significant differences among investigated treatments. High average number of microorganisms were determined in all treatments. Most abundant were TNB (log No 8,010) and ACT (log No 6,055) than FNG and AZB with log No 4,867 and log No 4,025, respectively. The highest DHA activity was measured in the FS of soybean in treatment 3. The application of nonsymbiotic bacteria in soybean production can alleviate initial plant growth and help the plant to better overcome different stress conditions caused by abiotic and biotic factors.

Keywords: bacteria, inoculation, soybean, microbial activity

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324 Preventive Effect of Three Kinds of Bacteriophages to Control Vibrio coralliilyticus Infection in Oyster Larvae

Authors: Hyoun Joong Kim, Jin Woo Jun, Sib Sankar Giri, Cheng Chi, Saekil Yun, Sang Guen Kim, Sang Wha Kim, Jeong Woo Kang, Se Jin Han, Se Chang Park

Abstract:

Vibrio corallilyticus is a well-known pathogen of coral. It is also infectious to a variety of shellfish species, including Pacific oyster (Crassostrea gigas) larvae. V. corallilyticus is remained to be a major constraint in marine bivalve aquaculture practice, especially in artificial seed production facility. Owing to the high mortality and contagious nature of the pathogen, large amount of antibiotics has been used for disease prevention and control. However, indiscriminate use of antibiotics may result in food and environmental pollution, and development of antibiotic resistant strains. Therefore, eco-friendly disease preventative measures are imperative for sustainable bivalve culture. The present investigation proposes the application of bacteriophage (phage) as an effective alternative method for controlling V. corallilyticus infection in marine bivalve hatcheries. Isolation of phages from sea water sample was carried out using drop or double layer agar methods. The host range, stability and morphology of the phage isolates were studied. In vivo phage efficacy to prevent V. corallilyticus infection in oyster larvae was also performed. The isolated phages, named pVco-5 and pVco-7 was classified as a podoviridae and pVco-14, was classified as a siphoviridae. Each phages were infective to four strains of seven V. corallilyticus strains tested. When oyster larvae were pre-treated with the phage before bacterial challenge, mortality of the treated oyster larvae was lower than that in the untreated control. This result suggests that each phages have the potential to be used as therapeutic agent for controlling V. corallilyticus infection in marine bivalve hatchery.

Keywords: bacteriophage, Vibrio coralliilyticus, Oyster larvae, mortality

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323 Biodegradation Ability of Polycyclic Aromatic Hydrocarbon (PAHs) Degrading Bacillus cereus Strain JMG-01 Isolated from PAHs Contaminated Soil

Authors: Momita Das, Sofia Banu, Jibon Kotoky

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Environmental contamination of natural resources with persistent organic pollutants is of great world-wide apprehension. Polycyclic aromatic hydrocarbons (PAHs) are among the organic pollutants, released due to various anthropogenic activities. Due to their toxic, carcinogenic and mutagenic properties, PAHs are of environmental and human concern. Presently, bioremediation has evolved as the most promising biotechnology for cleanup of such contaminants because of its economical and less cost effectiveness. In the present study, distribution of 16 USEPA priority PAHs was determined in the soil samples collected from fifteen different sites of Guwahati City, the Gateway of the North East Region of India. The total concentrations of 16 PAHs (Σ16 PAHs) ranged from 42.7-742.3 µg/g. Higher concentration of total PAHs was found more in the Industrial areas compared to all the sites (742.3 µg/g and 628 µg/g). It is noted that among all the PAHs, Naphthalene, Acenaphthylene, Anthracene, Fluoranthene, Chrysene and Benzo(a)Pyrene were the most available and contain the higher concentration of all the PAHs. Since microbial activity has been deemed the most influential and significant cause of PAH removal; further, twenty-three bacteria were isolated from the most contaminated sites using the enrichment process. These strains were acclimatized to utilize naphthalene and anthracene, each at 100 µg/g concentration as sole carbon source. Among them, one Gram-positive strain (JMG-01) was selected, and biodegradation ability and initial catabolic genes of PAHs degradation were investigated. Based on 16S rDNA analysis, the isolate was identified as Bacillus cereus strain JMG-01. Topographic images obtained using Scanning Electron Microscope (SEM) and Atomic Force Microscope (AFM) at scheduled time intervals of 7, 14 and 21 days, determined the variation in cell morphology during the period of degradation. AFM and SEM micrograph of biomass showed high filamentous growth leading to aggregation of cells in the form of biofilm with reference to the incubation period. The percentage degradation analysis using gas chromatography and mass analyses (GC-MS) suggested that more than 95% of the PAHs degraded when the concentration was at 500 µg/g. Naphthalene, naphthalene-2-methy, benzaldehyde-4-propyl, 1, 2, benzene di-carboxylic acid and benzene acetic acid were the major metabolites produced after degradation. Moreover, PCR experiments with specific primers for catabolic genes, ndo B and Cat A suggested that JMG-01 possess genes for PAHs degradation. Thus, the study concludes that Bacillus cereus strain JMG-01 has efficient biodegrading ability and can trigger the clean-up of PAHs contaminated soil.

Keywords: AFM, Bacillus cereus strain JMG-01, degradation, polycyclic aromatic hydrocarbon, SEM

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322 Effect of Cellulase Pretreatment for n-Hexane Extraction of Oil from Garden Cress Seeds

Authors: Boutemak Khalida, Dahmani Siham

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Garden cress (Lepidium Sativum L.) belonging to the family Brassicaceae, is edible growing annual herb. Its various parts (roots, leaves and seeds) have been used to treat various human ailments. Its seed extracts have been screened for various biological activities like hypotensive, antimicrobial, bronchodilator, hypoglycaemic and antianemic. The aim of the present study is to optimize the process parameters (cellulase concentration and incubation time) of enzymatic pre-treatment of the garden cress seeds and to evaluate the effect of cellulase pre-treatment of the crushed seeds on the oil yield, physico-chemical properties and antibacterial activity and comparing to non-enzymatic method. The optimum parameters of cellulase pre-treatment were as follows: cellulase of 0,1% w/w and incubation time of 2h. After enzymatic pre-treatment, the oil was extracted by n-hexane for 1.5 h, the oil yield was 4,01% for cellulase pre-treatment as against 10,99% in the control sample. The decrease in yield might be caused a result of mucilage. Garden cress seeds are covered with a layer of mucilage which gels on contact with water. At the same time, the antibacterial activity was carried out using agar diffusion method against 4 food-borne pathogens (Escherichia coli, Salmonella typhi,Staphylococcus aureus, Bacillus subtilis). The results showed that bacterial strains are very sensitive to the oil with cellulase pre-treatment. Staphylococcus aureus is extremely sensitive with the largest zone of inhibition (40 mm), Escherichia coli and salmonella typhi had a very sensitive to the oil with a zone of inhibition (26 mm). Bacillus subtilizes is averagely sensitive which gave an inhibition of 16 mm. But it does not exhibit sensivity to the oil without enzymatic pre-treatment with a zone inhibition (< 8 mm). Enzymatic pre-treatment could be useful for antimicrobial activity of the oil, and hold a good potential for use in food and pharmaceutical industries.

Keywords: Lepidium sativum L., cellulase, enzymatic pretreatment, antibacterial activity.

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321 The Effect of Extracts of 12 Local Medicinal Plants Against Uropathogenic Escherichia Coli

Authors: Hafida Merzouk

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Urinary tract infections are among the most serious public health issues in all age groups. Thus, the empirical therapy should based on local levels of resistance, as indicated in several studies from different countries, to effectively avoid the emergence of multidrug-resistant bacterial strains and recurrent infections. Numerous effective antibiotic treatments are available, but wouldbe ineffective for treating recurrent cystitis caused by a urinary tract infection, as well as the emergence of drug resistance. That iswhy the aim of this study was to highlight the antibacterial and the antioxidant activity of 11 medicinal plants used traditionally in Algeria against E. coli, the most responsible urinary tract infections. First, the extraction of total polyphenols with aqueous acetone showed variable yields. The highest yield was obtained by Asplenium trichomanes with 27%, followed by Petroselinum crispum and Ciannamomum cassia with an equal yield of 21%. Artemisia herba-alba gave the lowest yield (9%). The extracts of different plants showed variable contents of phenolic compounds. Reducing power and DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging activity revealed that most of the extracts studied had significant activity. The anti-free radical activity was very high in the extract of A splenium adiantum-nigrum compared with the other extracts studied, but Petroselinum crispum and Parietaria officinalis had the lowest reducing activity; Antibacterial activity was determined on E. coli strainsusing the diffusion, MICs (Minimum Inhibitory Concentrations) and MBCs (Minimum Bactericidal concentrations) methods. The strains tested were sensitive to most extracts studied, except Asplenium adiantum-nigrum extract, for which both strains showed resistance.

Keywords: E. coli, medicinal plants, phenolic compounds, urinary infections

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320 Synthesis of Highly Sensitive Molecular Imprinted Sensor for Selective Determination of Doxycycline in Honey Samples

Authors: Nadia El Alami El Hassani, Soukaina Motia, Benachir Bouchikhi, Nezha El Bari

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Doxycycline (DXy) is a cycline antibiotic, most frequently prescribed to treat bacterial infections in veterinary medicine. However, its broad antimicrobial activity and low cost, lead to an intensive use, which can seriously affect human health. Therefore, its spread in the food products has to be monitored. The scope of this work was to synthetize a sensitive and very selective molecularly imprinted polymer (MIP) for DXy detection in honey samples. Firstly, the synthesis of this biosensor was performed by casting a layer of carboxylate polyvinyl chloride (PVC-COOH) on the working surface of a gold screen-printed electrode (Au-SPE) in order to bind covalently the analyte under mild conditions. Secondly, DXy as a template molecule was bounded to the activated carboxylic groups, and the formation of MIP was performed by a biocompatible polymer by the mean of polyacrylamide matrix. Then, DXy was detected by measurements of differential pulse voltammetry (DPV). A non-imprinted polymer (NIP) prepared in the same conditions and without the use of template molecule was also performed. We have noticed that the elaborated biosensor exhibits a high sensitivity and a linear behavior between the regenerated current and the logarithmic concentrations of DXy from 0.1 pg.mL−1 to 1000 pg.mL−1. This technic was successfully applied to determine DXy residues in honey samples with a limit of detection (LOD) of 0.1 pg.mL−1 and an excellent selectivity when compared to the results of oxytetracycline (OXy) as analogous interfering compound. The proposed method is cheap, sensitive, selective, simple, and is applied successfully to detect DXy in honey with the recoveries of 87% and 95%. Considering these advantages, this system provides a further perspective for food quality control in industrial fields.

Keywords: doxycycline, electrochemical sensor, food control, gold nanoparticles, honey, molecular imprinted polymer

Procedia PDF Downloads 317
319 Infection Profile of Patients Undergoing Autologous Bone Marrow Transplantation in Tabriz, Iran

Authors: Naser Shagerdi Esmaeli, Mohsen Hamidpour

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Background and Objective: Hematopoietic stem cell transplantation (HSCT) has been widely used for treating oncological and hematological diseases. Although HSCT has helped to improve patient survival, the risk of developing an infection during hospitalization is an important cause of morbidity and mortality. This study aimed to analyze the infection profile during hospitalization and the associated risk factors among patients undergoing autologous HSCT at the University Hospital, Shahid Ghazi Tabatabaei Hospital, Tabriz, Iran. Subjects and Methods: This was a cross-sectional study on patients undergoing autologous HSCT at a public university hospital. Methods: Patients with febrile neutropenia between 2015 and 2018 were retrospectively evaluated regarding their infection profile and associated risk factors. This survey included: bacterial culture and blood culture on specific media. Results: Infection occurred in 57.2% of 56 patients with febrile neutropenia. The main source of infection was the central venous catheter (25.9%). Infection was chiefly due to Gram-positive bacteria, although Gram-negative-related infections were more severe and caused a higher death rate. Sex, age, skin color, nutritional status, and underlying disease were not associated with the development of infection. Patients with severe mucositis (Grades III and IV) had a higher infection rate (P < 0.001). Patients who developed pulmonary complications during hospitalization had higher infection rates (P = 0.002). Infection was the main cause of death (57.1%) in the study sample. Conclusion: Strategies aimed at reducing infection-related mortality rates among patients undergoing autologous HSCT are necessary.

Keywords: hematopoietic stem cell, autologous bone marrow transplantation, infection profile, tabriz, Iran

Procedia PDF Downloads 120
318 Application of Nanoparticles on Surface of Commercial Carbon-Based Adsorbent for Removal of Contaminants from Water

Authors: Ahmad Kayvani Fard, Gordon Mckay, Muataz Hussien

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Adsorption/sorption is believed to be one of the optimal processes for the removal of heavy metals from water due to its low operational and capital cost as well as its high removal efficiency. Different materials have been reported in literature as adsorbent for heavy metal removal in waste water such as natural sorbents, organic polymers (synthetic) and mineral materials (inorganic). The selection of adsorbents and development of new functional materials that can achieve good removal of heavy metals from water is an important practice and depends on many factors, such as the availability of the material, cost of material, and material safety and etc. In this study we reported the synthesis of doped Activated carbon and Carbon nanotube (CNT) with different loading of metal oxide nanoparticles such as Fe2O3, Fe3O4, Al2O3, TiO2, SiO2 and Ag nanoparticles and their application in removal of heavy metals, hydrocarbon, and organics from waste water. Commercial AC and CNT with different loadings of mentioned nanoparticle were prepared and effect of pH, adsorbent dosage, sorption kinetic, and concentration effects are studied and optimum condition for removal of heavy metals from water is reported. The prepared composite sorbent is characterized using field emission scanning electron microscopy (FE-SEM), high transmission electron microscopy (HR-TEM), thermogravimetric analysis (TGA), X-ray diffractometer (XRD), the Brunauer, Emmett and Teller (BET) nitrogen adsorption technique, and Zeta potential. The composite materials showed higher removal efficiency and superior adsorption capacity compared to commercially available carbon based adsorbent. The specific surface area of AC increased by 50% reaching up to 2000 m2/g while the CNT specific surface area of CNT increased by more than 8 times reaching value of 890 m2/g. The increased surface area is one of the key parameters along with surface charge of the material determining the removal efficiency and removal efficiency. Moreover, the surface charge density of the impregnated CNT and AC have enhanced significantly where can benefit the adsorption process. The nanoparticles also enhance the catalytic activity of material and reduce the agglomeration and aggregation of material which provides more active site for adsorbing the contaminant from water. Some of the results for treating wastewater includes 100% removal of BTEX, arsenic, strontium, barium, phenolic compounds, and oil from water. The results obtained are promising for the use of AC and CNT loaded with metal oxide nanoparticle in treatment and pretreatment of waste water and produced water before desalination process. Adsorption can be very efficient with low energy consumption and economic feasibility.

Keywords: carbon nanotube, activated carbon, adsorption, heavy metal, water treatment

Procedia PDF Downloads 234
317 Surface Defect-engineered Ceo₂−x by Ultrasound Treatment for Superior Photocatalytic H₂ Production and Water Treatment

Authors: Nabil Al-Zaqri

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Semiconductor photocatalysts with surface defects display incredible light absorption bandwidth, and these defects function as highly active sites for oxidation processes by interacting with the surface band structure. Accordingly, engineering the photocatalyst with surface oxygen vacancies will enhance the semiconductor nanostructure's photocatalytic efficiency. Herein, a CeO2₋ₓ nanostructure is designed under the influence of low-frequency ultrasonic waves to create surface oxygen vacancies. This approach enhances the photocatalytic efficiency compared to many heterostructures while keeping the intrinsiccrystal structure intact. Ultrasonic waves induce the acoustic cavitation effect leading to the dissemination of active elements on the surface, which results in vacancy formation in conjunction with larger surface area and smaller particle size. The structural analysis of CeO₂₋ₓ revealed higher crystallinity, as well as morphological optimization, and the presence of oxygen vacancies is verified through Raman, X-rayphotoelectron spectroscopy, temperature-programmed reduction, photoluminescence, and electron spinresonance analyses. Oxygen vacancies accelerate the redox cycle between Ce₄+ and Ce₃+ by prolongingphotogenerated charge recombination. The ultrasound-treated pristine CeO₂ sample achieved excellenthydrogen production showing a quantum efficiency of 1.125% and efficient organic degradation. Ourpromising findings demonstrated that ultrasonic treatment causes the formation of surface oxygenvacancies and improves photocatalytic hydrogen evolution and pollution degradation. Conclusion: Defect engineering of the ceria nanoparticles with oxygen vacancies was achieved for the first time using low-frequency ultrasound treatment. The U-CeO₂₋ₓsample showed high crystallinity, and morphological changes were observed. Due to the acoustic cavitation effect, a larger surface area and small particle size were observed. The ultrasound treatment causes particle aggregation and surface defects leading to oxygen vacancy formation. The XPS, Raman spectroscopy, PL spectroscopy, and ESR results confirm the presence of oxygen vacancies. The ultrasound-treated sample was also examined for pollutant degradation, where 1O₂was found to be the major active species. Hence, the ultrasound treatment influences efficient photocatalysts for superior hydrogen evolution and an excellent photocatalytic degradation of contaminants. The prepared nanostructure showed excellent stability and recyclability. This work could pave the way for a unique post-synthesis strategy intended for efficient photocatalytic nanostructures.

Keywords: surface defect, CeO₂₋ₓ, photocatalytic, water treatment, H₂ production

Procedia PDF Downloads 141
316 Development of Hydrophilic Materials for Nanofiltration Membrane Achieving Dual Resistance to Fouling and Chlorine

Authors: Xi Quan Cheng, Yan Chao Xu, Xu Jiang, Lu Shao, Cher Hon Lau

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A hydrophilic thin-film-composite (TFC) nanofiltration (NF) membrane has been developed through the interfacial polymerization (IP) of amino-functional polyethylene glycol (PEG) and trimesoyl chloride. The selective layer is formed on a polyethersulfone (PES) support that is characterized using FTIR, XPS and SEM, and is dependent on monomer immersion duration, and the concentration of monomers and additives. The higher hydrophilicity alongside the larger pore size of the PEG-based selective layer is the key to a high water flux of 66.0 L m-2 h-1 at 5.0 bar. With mean pore radius of 0.42 nm and narrow pore size distribution, the MgSO4 rejections of the PEG based PA TFC NF membranes can reach up to 80.2 %. The hydrophilic PEG based membranes shows positive charged since the isoelectric points range from pH=8.9 to pH=9.1 and the rejection rates for different salts of the novel membranes are in the order of R(MgCl2)>R(MgSO4)>R(NaCl)>R(Na2SO4). The pore sizes and water permeability of these membranes are tailored by varying the molecular weight and molecular architecture of amino-functional PEG. Due to the unique structure of the selective layer of the PEG based membranes consisting of saturated aliphatic construction unit (CH2-CH2-O), the membranes demonstrate dual resistance to fouling and chlorine. The membranes maintain good salt rejections and high water flux of PEG based membranes after treatment by 2000 ppm NaClO for 24 hours. Interestingly, the PEG based membranes exhibit excellent fouling resistance with a water flux recovery of 90.2 % using BSA as a model molecule. More importantly, the hydrophilic PEG based NF membranes have been exploited to separate several water soluble antibiotics (such as tobramycin, an aminoglycoside antibiotic applied in the treatment of various types of bacterial infections), showing excellent performance in concentration or removal of antibioics.

Keywords: nanofiltration, antibiotic separation, hydrophilic membrane, high flux

Procedia PDF Downloads 318
315 Plant Growth, Symbiotic Performance and Grain Yield of 63 Common Bean Genotypes Grown Under Field Conditions at Malkerns Eswatini

Authors: Rotondwa P. Gunununu, Mustapha Mohammed, Felix D. Dakora

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Common bean is the most importantly high protein grain legume grown in Southern Africa for human consumption and income generation. Although common bean can associate with rhizobia to fix N₂ for bacterial use and plant growth, it is reported to be a poor nitrogen fixer when compared to other legumes. N₂ fixation can vary with legume species, genotype and rhizobial strain. Therefore, screening legume germplasm can reveal rhizobia/genotype combinations with high N₂-fixing efficiency for use by farmers. This study assessed symbiotic performance and N₂ fixation in 63 common bean genotypes under field conditions at Malkerns Station in Eswatini, using the ¹⁵N natural abundance technique. The shoots of common bean genotypes were sampled at a pod-filling stage, oven-dried (65oC for 72h), weighed, ground into a fine powder (0.50 mm sieve), and subjected to ¹⁵N/¹⁴N isotopic analysis using mass spectrometry. At maturity, plants from the inner rows were harvested for the determination of grain yield. The results revealed significantly higher modulation (p≤0.05) in genotypes MCA98 and CIM-RM01-97-8 relative to the other genotypes. Shoot N concentration was highest in genotype MCA 98, followed by KAB 10 F2.8-84, with most genotypes showing shoot N concentrations below 2%. Percent N derived from atmospheric N₂ fixation (%Ndfa) differed markedly among genotypes, with CIM-RM01-92-3 and DAB 174, respectively, recording the highest values of 66.65% and 66.22 % N derived from fixation. There were also significant differences in grain yield, with CIM-RM02-79-1 producing the highest yield (3618.75 kg/ha). These results represent an important contribution in the profiling of symbiotic functioning of common bean germplasm for improved N₂ fixation.

Keywords: nitrogen fixation, %Ndfa, ¹⁵N natural abundance, grain yield

Procedia PDF Downloads 219
314 Comparative Correlation Investigation of Polynuclear Aromatic Hydrocarbons (PAHs) in Soils of Different Land Uses: Sources Evaluation Perspective

Authors: O. Onoriode Emoyan, E. Eyitemi Akporhonor, Charles Otobrise

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Polycyclic Aromatic Hydrocarbons (PAHs) are formed mainly as a result of incomplete combustion of organic materials during industrial, domestic activities or natural occurrence. Their toxicity and contamination of terrestrial and aquatic ecosystem have been established. Though with limited validity index, previous research has focused on PAHs isomer pair ratios of variable physicochemical properties in source identification. The objective of this investigation was to determine the empirical validity of Pearson correlation coefficient (PCC) and cluster analysis (CA) in PAHs source identification along soil samples of different land uses. Therefore, 16 PAHs grouped as endocrine disruption substances (EDSs) were determined in 10 sample stations in top and sub soils seasonally. PAHs was determined the use of Varian 300 gas chromatograph interfaced with flame ionization detector. Instruments and reagents used are of standard and chromatographic grades respectively. PCC and CA results showed that the classification of PAHs along kinetically and thermodyanamically-favoured and those derived directly from plants product through biologically mediated processes used in source signature is about the predominance PAHs are likely to be. Therefore the observed PAHs in the studied stations have trace quantities of the vast majority of the sixteen un-substituted PAHs which may ultimately inhabit the actual source signature authentication. Type and extent of bacterial metabolism, transformation products/substrates, and environmental factors such as: salinity, pH, oxygen concentration, nutrients, light intensity, temperature, co-substrates and environmental medium are hereby recommended as factors to be considered when evaluating possible sources of PAHs.

Keywords: comparative correlation, kinetically and thermodynamically-favored PAHs, pearson correlation coefficient, cluster analysis, sources evaluation

Procedia PDF Downloads 422
313 Escherichia Coli Producing Extended-spectrum Beta-lactamase (ESBL) at the Tambacounda Regional Hospital Center (CHRT), Senegal

Authors: Assane Dieng, Abass Sarr, Gora Lo, Awa Ba, Halimatou Diop Ndiaye, Makhtar Camara

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Introduction: Escherchia coli is one of the main beta-lactamase-producing bacteria. In Senegal, we are witnessing a weakness in the technical platform and a deficit in qualified personnel in laboratories, especially in rural areas. This is why there is little updated data on antibiotic bacterial resistance in rural areas. The objective is to study infections with Escherichia coli producing extended-spectrum beta-lactamase (ESBL) at the Tambacounda Regional Hospital Center (CHRT), Senegal. Methodology: Samples were taken from patients consulted at the Tambacounda Regional Hospital Center (CHRT) from January to October 2024. The search for Escherichia coli was carried out by determining morphological, biochemical and cultural characteristics. The antibiogram was performed using the Mueller-Hinton (MH) agar diffusion method following the recommendations of the Antibiogram Committee of the French Society of Microbiology (CA SFM). ESBL production was detected using the double synergy method Results: A total of 287 germs were isolated during the study period, including 96 strains of Escherichia coli, 47 of which produced a broad-spectrum beta-lactamase. ESBL-producing E. coli strains were more common in men (61.2%). The most affected age groups were [60-79 years] and [20-39 years] with respective rates of 36.7% and 27%. ESBL-producing E. coli strains were isolated mainly in non-hospitalized patients (79%). Beta-lactamase-producing E. coli strains were mainly resistant to tetracycline (95.7%), norfloxacin (91.5%) and ciprofloxacin (87.2%).Conclusion: ESBL-producing E. coli strains are also present in rural areas Thus, to have more comprehensive data, qualified personnel as well as a high-level technical platform should be available in these laboratories.

Keywords: E. coli, beta-lactamase, rural area, non hospitalized

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312 Effect of Chemical Mutagen on Seeds Germination of Lima Bean

Authors: G. Ultanbekova, Zh. Suleimenova, Zh. Rakhmetova, G. Mombekova, S. Mantieva

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Plant Growth Promoting Rhizobacteria (PGPR) are a group of free-living bacteria that colonize the rhizosphere, enhance plant growth of many cereals and other important agricultural crops and protect plants from disease and abiotic stresses through a wide variety of mechanisms. The use of PGPR has been proven to be an environmentally sound way of increasing crop yields by facilitating plant growth. In the present study, strain improvement of PGPR isolates were carried out by chemical mutagenesis for the improvement of growth and yield of lima bean. Induced mutagenesis is widely used for the selection of microorganisms producing biologically active substances and further improving their activities. Strain improvement is usually done by classical mutagenesis which involves exposing the microbes to chemical or physical mutagens. The strains of Pseudomonas putida 4/1, Azotobacter chroococcum Р-29 and Bacillus subtilis were subjected to mutation process for strain improvement by treatment with a chemical agent (sodium nitrite) to cause mutation and were observed for its consequent action on the seeds germination and plant growth of lima bean (Phaseolus lunatus). Bacterial mutant strains of Pseudomonas putida M-1, Azotobacter chroococcum M-1 and Bacillus subtilis M-1, treated with sodium nitrite in the concentration of 5 mg/ml for 120 min, were found effective to enhance the germination of lima bean seeds compared to parent strains. Moreover, treatment of the lima bean seeds with a mutant strain of Bacillus subtilis M-1 had a significant stimulation effect on plant growth. The length of the stems and roots of lima bean treated with Bacillus subtilis M-1 increased significantly in comparison with parent strain in 1.6 and 1.3 times, respectively.

Keywords: chemical mutagenesis, germination, kidney bean, plant growth promoting rhizobacteria (PGPR)

Procedia PDF Downloads 199
311 Modification of Escherichia coli PtolT Expression Vector via Site-Directed Mutagenesis

Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey

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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.

Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression

Procedia PDF Downloads 376
310 Assessment of Some Biological Activities of Methanolic Crude Extract from Polygonum maritimum L.

Authors: Imad Abdelhamid El-Haci, Wissame Mazari, Fayçal Hassani, Fawzia Atik Bekkara

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Much attention has been paid to the antioxidants, which are expected to prevent food and living systems from peroxidative damage. Incorporation of synthetic antioxidants in food products is under strict regulation due to the potential health hazards caused by such compounds. The use of plants as traditional health remedies is very popular and important for 80% of the world’s population in African, Asian, Latin America and Middle Eastern Countries. Their use is reported to have minimal side effects. In recent years, pharmaceutical companies have spent considerable time and money in developing therapeutics based upon natural products extracted from plants. In other part, due to the continuous emergence of antibiotic-resistant strains there is continual demand for new antibiotics. Chemical compounds from medicinal plant especially are targeted by many researches. In this light, genus Polygonum (Polygonaceae), comprising about 45 genera (300 species), is distributed worldwide, mostly in north temperate regions. They have been reported to have uses in traditional medicine, such as anti-inflammation, promoting blood circulation, dysentery, diuretic, haemorrhage and many other uses. In our study, Polygonum maritimum (from Algerian coast) was extracted with 80% methanol to obtain a crude extract. P. maritimum extract (PME) had a very high content of total phenol, which was 352.49 ± 18.03 mg/g dry weight, expressed as gallic acid equivalent. PME exhibited excellent antioxidant activity, as measured using DPPH and H2O2 scavenging assays. It also showed a high antibacterial activity against gram positive bacterial strains: Bacillus cereus, Bacillus subtilis and Staphylococcus aureus with an MIC 0,12 mg/mL.

Keywords: Polygonum maritimum, crude extract, antioxidant activity, antibacterial activity

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309 The Potential of Edaphic Algae for Bioremediation of the Diesel-Contaminated Soil

Authors: C. J. Tien, C. S. Chen, S. F. Huang, Z. X. Wang

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Algae in soil ecosystems can produce organic matters and oxygen by photosynthesis. Heterocyst-forming cyanobacteria can fix nitrogen to increase soil nitrogen contents. Secretion of mucilage by some algae increases the soil water content and soil aggregation. These actions will improve soil quality and fertility, and further increase abundance and diversity of soil microorganisms. In addition, some mixotrophic and heterotrophic algae are able to degrade petroleum hydrocarbons. Therefore, the objectives of this study were to analyze the effects of algal addition on the degradation of total petroleum hydrocarbons (TPH), diversity and activity of bacteria and algae in the diesel-contaminated soil under different nutrient contents and frequency of plowing and irrigation in order to assess the potential bioremediation technique using edaphic algae. The known amount of diesel was added into the farmland soil. This diesel-contaminated soil was subject to five settings, experiment-1 with algal addition by plowing and irrigation every two weeks, experiment-2 with algal addition by plowing and irrigation every four weeks, experiment-3 with algal and nutrient addition by plowing and irrigation every two weeks, experiment-4 with algal and nutrient addition by plowing and irrigation every four weeks, and the control without algal addition. Soil samples were taken every two weeks to analyze TPH concentrations, diversity of bacteria and algae, and catabolic genes encoding functional degrading enzymes. The results show that the TPH removal rates of five settings after the two-month experimental period were in the order: experiment-2 > expermient-4 > experiment-3 > experiment-1 > control. It indicated that algal addition enhanced the degradation of TPH in the diesel-contaminated soil, but not for nutrient addition. Plowing and irrigation every four weeks resulted in more TPH removal than that every two weeks. The banding patterns of denaturing gradient gel electrophoresis (DGGE) revealed an increase in diversity of bacteria and algae after algal addition. Three petroleum hydrocarbon-degrading algae (Anabaena sp., Oscillatoria sp. and Nostoc sp.) and two added algal strains (Leptolyngbya sp. and Synechococcus sp.) were sequenced from DGGE prominent bands. The four hydrocarbon-degrading bacteria Gordonia sp., Mycobacterium sp., Rodococcus sp. and Alcanivorax sp. were abundant in the treated soils. These results suggested that growth of indigenous bacteria and algae were improved after adding edaphic algae. Real-time polymerase chain reaction results showed that relative amounts of four catabolic genes encoding catechol 2, 3-dioxygenase, toluene monooxygenase, xylene monooxygenase and phenol monooxygenase were appeared and expressed in the treated soil. The addition of algae increased the expression of these genes at the end of experiments to biodegrade petroleum hydrocarbons. This study demonstrated that edaphic algae were suitable biomaterials for bioremediating diesel-contaminated soils with plowing and irrigation every four weeks.

Keywords: catabolic gene, diesel, diversity, edaphic algae

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308 Development of Fluorescence Resonance Energy Transfer-Based Nanosensor for Measurement of Sialic Acid in vivo

Authors: Ruphi Naz, Altaf Ahmad, Mohammad Anis

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Sialic acid (5-Acetylneuraminic acid, Neu5Ac) is a common sugar found as a terminal residue on glycoconjugates in many animals. Humans brain and the central nervous system contain the highest concentration of sialic acid (as N-acetylneuraminic acid) where these acids play an important role in neural transmission and ganglioside structure in synaptogenesis. Due to its important biological function, sialic acid is attracting increasing attention. To understand metabolic networks, fluxes and regulation, it is essential to be able to determine the cellular and subcellular levels of metabolites. Genetically-encoded fluorescence resonance energy transfer (FRET) sensors represent a promising technology for measuring metabolite levels and corresponding rate changes in live cells. Taking this, we developed a genetically encoded FRET (fluorescence resonance energy transfer) based nanosensor to analyse the sialic acid level in living cells. Sialic acid periplasmic binding protein (sia P) from Haemophilus influenzae was taken and ligated between the FRET pair, the cyan fluorescent protein (eCFP) and Venus. The chimeric sensor protein was expressed in E. coli BL21 (DE3) and purified by affinity chromatography. Conformational changes in the binding protein clearly confirmed the changes in FRET efficiency. So any change in the concentration of sialic acid is associated with the change in FRET ratio. This sensor is very specific to sialic acid and found stable with the different range of pH. This nanosensor successfully reported the intracellular level of sialic acid in bacterial cell. The data suggest that the nanosensors may be a versatile tool for studying the in vivo dynamics of sialic acid level non-invasively in living cells

Keywords: nanosensor, FRET, Haemophilus influenzae, metabolic networks

Procedia PDF Downloads 133
307 Organic Permeation Properties of Hydrophobic Silica Membranes with Different Functional Groups

Authors: Sadao Araki, Daisuke Gondo, Satoshi Imasaka, Hideki Yamamoto

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The separation of organic compounds from aqueous solutions is a key technology for recycling valuable organic compounds and for the treatment of wastewater. The wastewater from chemical plants often contains organic compounds such as ethyl acetate (EA), methylethyl ketone (MEK) and isopropyl alcohol (IPA). In this study, we prepared hydrophobic silica membranes by a sol-gel method. We used phenyltrimethoxysilane (PhTMS), ethyltrimethoxysilan (ETMS), Propyltrimethoxysilane (PrTMS), N-butyltrimethoxysilane (BTMS), N-Hexyltrimethoxysilane (HTMS) as silica sources to introduce each functional groups on the membrane surface. Cetyltrimethyl ammonium bromide (CTAB) was used as a molecular template to create suitable pore that enable the permeation of organic compounds. These membranes with five different functional groups were characterized by SEM, FT-IR, and permporometry. Thicknesses and pore diameters of silica layer for all membrane were about 1.0 μm and about 1 nm, respectively. In other words, functional groups had an insignificant effect on the membrane thicknesses and the formation of the pore by CTAB. We confirmed the effect of functional groups on the flux and separation factor for ethyl acetate (EA), methyl ethyl ketone, acetone and 1-butanol (1-BtOH) /water mixtures. All membranes showed a high flux for ethyl acetate compared with other compounds. In particular, the hydrophobic silica membrane prepared by using BTMS showed 0.75 kg m-2 h-1 of flux for EA. For all membranes, the fluxes of organic compounds showed the large values in the order corresponding to EA > MEK > acetone > 1-BtOH. On the other hand, carbon chain length of functional groups among ETMS, PrTMS, BTMS, PrTMS and HTMS did not have a major effect on the organic flux. Although we confirmed the relationship between organic fluxes and organic molecular diameters or fugacity of organic compounds, these factors had a low correlation with organic fluxes. It is considered that these factors affect the diffusivity. Generally, permeation through membranes is based on the diffusivity and solubility. Therefore, it is deemed that organic fluxes through these hydrophobic membranes are strongly influenced by solubility. We tried to estimate the organic fluxes by Hansen solubility parameter (HSP). HSP, which is based on the cohesion energy per molar volume and is composed of dispersion forces (δd), intermolecular dipole interactions (δp), and hydrogen-bonding interactions (δh), has recently attracted attention as a means for evaluating the resolution and aggregation behavior. Evaluation of solubility for two substances can be represented by using the Ra [(MPa)1/2] value, meaning the distance of HSPs for both of substances. A smaller Ra value means a higher solubility for each substance. On the other hand, it can be estimated that the substances with large Ra value show low solubility. We established the correlation equation, which was based on Ra, of organic flux at low concentrations of organic compounds and at 295-325 K.

Keywords: hydrophobic, membrane, Hansen solubility parameter, functional group

Procedia PDF Downloads 379
306 Comparison of Bactec plus Blood Culture Media to BacT/Alert FAN plus Blood Culture Media for Identification of Bacterial Pathogens in Clinical Samples Containing Antibiotics

Authors: Recep Kesli, Huseyin Bilgin, Ela Tasdogan, Ercan Kurtipek

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Aim: The aim of this study was to compare resin based Bactec plus aerobic/anaerobic blood culture bottles (Becton Dickinson, MD, USA) and polymeric beads based BacT/Alert FA/FN plus blood culture bottles (bioMerieux, NC, USA) in terms of microorganisms recovery rates and time to detection (TTD) in the patients receiving antibiotic treatment. Method: Blood culture samples were taken from the patients who admitted to the intensive care unit and received antibiotic treatment. Forty milliliters of blood from patients were equally distributed into four types of bottles: Bactec Plus aerobic, Bactec Plus anaerobic, BacT/Alert FA Plus, BacT/Alert FN Plus. Bactec Plus and BacT/Alert Plus media were compared to culture recovery rates and TTD. Results: Blood culture samples were collected from 382 patients hospitalized in the intensive care unit and 245 patients who were diagnosed as having bloodstream infections were included in the study. A total of 1528 Bactec Plus aerobic, Bactec Plus anaerobic, BacT/Alert FA Plus, BacT/Alert FN Plus blood culture bottles analyzed and 176, 144, 154, 126 bacteria or fungi were isolated, respectively. Gram-negative and gram-positive bacteria were significantly more frequently isolated in the resin-based Bactec Plus bottles than in the polymeric beads based BacT/Alert Plus bottles. The Bactec Plus and BacT/Alert Plus media recovery rates were similar for fungi and anaerobic bacteria. The mean TTDs in the Bactec Plus bottles were shorter than those in the BacT/Alert Plus bottles regardless of the microorganisms. Conclusion: The results of this study showed that resin-containing media is a reliable and time-saving tool for patients who are receiving antibiotic treatment due to sepsis in the intensive care unit.

Keywords: Bactec Plus, BacT/Alert Plus, blood culture, antibiotic

Procedia PDF Downloads 146