Search results for: bacterial aggregation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1435

Search results for: bacterial aggregation

985 Bacteriological Quality and Physicochemical Water Beaches of the City of Annaba (Mediterranean Sea)

Authors: Wahiba Boudraa, Farah Chettibbi, Meriem Aberkane, Fatma Djamaa, Moussa Houhamdi

Abstract:

The intensity of human activities in regions surrounding the Mediterranean Sea always has a strong long-term environmental impact resulting in coastal and marine degradation, as well as an aggravated risk of more serious damage. The available data on water quality show that most water resources in Algeria are polluted by uncontrolled discharges from municipal sewage and untreated industrial effluents. Annaba is a coastal town in Algeria; The Gulf of Annaba, responds to these changes as it receives the continental inputs and urban waste, industrial without prior treatment of a highly industrialized and urbanized city, subject to the same environmental problems that know the rest of the Algerian coast. In later year, the beaches of bacterial enumeration process waters showed relatively high levels of bacterial indicators of fecal contamination (group D streptococci, total and fecal coliforms), which reflect the risks to people attending these beaches. During the twelve months of our study, we isolated from three beaches in the city of Annaba (St. Cloud, El-Kettara, and Djenane El Bey) a number of pathogenic microorganisms considered, namely: Salmonella, Aeromonas, Citrobacter, Yersinia, Enterococcus, and E.coli. The microbial count revealed elevated levels of coliform bacteria, fecal coliforms and fecal streptococci quite high especially in urban beaches (St. Cloud and El-Kettara). They are widely popular during the summer by many vacationers. For the physico-chemical parameters, there exist some weak values which increase during the pluvial period, hivernal and festival saison. These values remain, nevertheless, weak to be able to cause an organic or metallic pollution.

Keywords: quality microbiology, pollution of water, fecal contamination, physico-chemistry, beaches of Annaba city, Algeria.

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984 Polyacrylate Modified Copper Nanoparticles with Controlled Size

Authors: Robert Prucek, Aleš Panáček, Jan Filip, Libor Kvítek, Radek Zbořil

Abstract:

The preparation of Cu nanoparticles (NPs) through the reduction of copper ions by sodium borohydride in the presence of sodium polyacrylate with a molecular weight of 1200 is reported. Cu NPs were synthesized at a concentration of copper salt equal to 2.5, 5, and 10 mM, and at a molar ratio of copper ions and monomeric unit of polyacrylate equal to 1:2. The as-prepared Cu NPs have diameters of about 2.5–3 nm for copper concentrations of 2.5 and 5 mM, and 6 nm for copper concentration of 10 mM. Depending on the copper salt concentration and concentration of additionally added polyacrylate to Cu particle dispersion, primarily formed NPs grow through the process of aggregation and/or coalescence into clusters and/or particles with a diameter between 20–100 nm. The amount of additionally added sodium polyacrylate influences the stability of Cu particles against air oxidation. The catalytic efficiency of the prepared Cu particles for the reduction of 4-nitrophenol is discussed.

Keywords: copper, nanoparticles, sodium polyacrylate, catalyst, 4-nitrophenol

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983 Analysis of Chemical Composition of Raw Milk in Some Farms Animals in El Khoms, Libya

Authors: Dukali Abujnah

Abstract:

This study was undertaken to knowledge the chemical composition of some farm animals (cows ) by using ultrasonic milk analyzer. This method was used first time in our country in the field of milk hygiene to determine and comparison the composition percent of raw milk for farm animals as attempt to protect the health of consumers 240 raw milk samples for each species of animals included in this study for two different season of year first (Novembe 2014 - January 2015) , second from (May – Novembe2015) .The chemical analysis. In this way, a range of building suitable to put in the Libyan market product structures, and milk, for use as a new product or exposure to treatment such as pasteurized milk and long-life may be wider study project..Controls have been implemented in operating in Libya, companies which are part of the business created by the Libyan state. It was created only after the companies that made many of the studies and research carried out on areas of Libya which it was possible to intervene. Although troops spent in recent years, companies have not yet found their balance and milk production has not yet come to those set forth in the action plans, due to problems of various types that will be discussed in this study. We believe that these data have been taken in four years if analyzed properly, can provide more accurate information to farmers in their companies want to take dairy cattle breeding. The total bacterial count was 76 and 38 cfu/ml for locally produced and imported UHT milk respectively. Coliforms were negative for all examined locally produce and imported samples. E. coli were not isolated from any of the examined UHT samples. Aerobic spore-formers count mean count was 49 and 27 cfu/ml for both examined samples respectively. Obtained results show that most of the locally produced UHT milk does not satisfy the suggested Libyan Standards for UHT milk for fat% as well as total bacterial count/ml. Economical and public health importance of the obtained results were discussed, and control measures for improving the quality of UHT locally made milk were also discussed.

Keywords: locally, UHT milks, farms animals, raw milk analysis milk

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982 Enhanced Anti-Obesity Effect of Soybean by Fermentation with Lactobacillus plantarum P1201 in 3T3-L1 Adipocyte

Authors: Chengliang Xie, Jinhyun Ryu, Hyun Joon Kim, Gyeong Jae Cho, Wan Sung Choi, Sang Soo Kang, Kye Man Cho, Dong Hoon Lee

Abstract:

Obesity has become a global health problem and a source of major metabolic diseases like type-2 diabetes, hypertension, heart disease, nonalcoholic fatty liver and cancer. Synthetic anti-obesity drugs are effective but very costly and with undesirable side effects, so natural products such as soybean are needed as an alternative for obesity treatment. Lactobacillus Plantarum P1201is a probiotic bacterial strain reported to produce conjugated linoleic acid (CLA) and increase the ratio of aglycone-isoflavone of soybean, both of which have anti-obesity effect. In this study, the anti-obesity effect of the fermented soybean extract with P1201 (FSE) will be evaluated compared with that of the soybean extract (SE) by 3T3-L1 cells as an in vitro model of adipogenesis. 3T3-L1 cells were treated with SE and FSE during the nine days of the differentiation, lipid accumulation was evaluated by oil-red staining and triglyceride content and the mRNA expression level of adipogenic or lipogenic genes were analyzed by RT-PCR and qPCR. The results showed that formation of lipid droplets in differentiated 3T3-L1 cells was inhibited and triglyceride content was reduced by 23.1% after treated with 1000 μg/mL of FSE compared with control. For SE-treated groups, no delipidating effect was observed. The effect of FSE on adipogenesis inhibition can be attributed to the down-regulation of mRNA expressionof CCAAT/enhancer binding protein (C/EBP-α), lipoprotein lipase (LPL), adiponectin, adipocyte fatty acid-binding protein (aP2), fatty acid synthesis (FAS) and CoA carboxylase (ACC). Our results demonstrated that the anti-obesity effect of soybean can be improved by fermentation with P1201, and P1201can be used as a potential probiotic bacterial strain to produce natural anti-obesity food.

Keywords: fermentation, Lactobacillus plantarum P1201, obesity, soybean

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981 Potential of Water Purification of Turbid Surface Water Sources in Remote Arid and Semi-Arid Rural Areas of Rajasthan by Moringa Oleifera (Drumstick) Tree Seeds

Authors: Pomila Sharma

Abstract:

Rajasthan is among regions with greatest climate sensitivity and lowest adaptive capabilities. In many parts of the Rajasthan surface water which can be highly turbid and contaminated with fecal coliform bacteria is used for drinking purposes. The majority rely almost exclusively upon traditional sources of highly turbid and untreated pathogenic surface water for their domestic water needs. In many parts of rural areas of Rajasthan, it is still difficult to obtain clean water, especially remote habitations with no groundwater due to quality issues or depletion and limited feasibility to connect with surface water schemes due to low density of population in these areas to justify large infrastructure investment. The most viable sources are rain water harvesting, community managed open wells, private wells, ponds and small-scale irrigation reservoirs have often been the main traditional sources of rural drinking water. Turbidity is conventionally removed by treating the water with expensive chemicals. This study has to investigate the use of crushed seeds from the tree Moringa oleifera (drumstick) as a natural alternative to conventional coagulant chemicals. The use of Moringa oleifera seed powder can produce potable water of higher quality than the original source. Moringa oleifera a native species of northern India, the tree is now grown extensively throughout the tropics and found in many countries of Africa, Asia & South America. The seeds of tree contains significant quantities of low molecular weight, water soluble proteins which carries the positive charge when the crushed seeds are added to water. This protein binds in raw water with negatively charged turbid water with bacteria, clay, algae, etc. Under proper mixing, these particles make flocks, which may be left to settle by gravity or be removed by filtration. Using Moringa oleifera as a replacement coagulation in such surface sources of arid and semi-arid areas can meet the need for water purification in remote places of Rajasthan state of India. The present study accesses to find out laboratory based investigation of the effect of seeds of Moringa tree on its coagulation effectiveness (purification) using turbid water samples of surface source of the Rajasthan state. In this study, moringa seed powder showed that filtering with seed powder may diminish water pollution and bacterial counts. Results showed Moringa oleifera seeds coagulate 90-95% of turbidity and color efficiently leading to an aesthetically clear supernatant & reduced about 85-90% of bacterial load reduction in samples.

Keywords: bacterial load, coagulant, turbidity, water purification

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980 Time Synchronization between the eNBs in E-UTRAN under the Asymmetric IP Network

Authors: M. Kollar, A. Zieba

Abstract:

In this paper, we present a method for a time synchronization between the two eNodeBs (eNBs) in E-UTRAN (Evolved Universal Terrestrial Radio Access) network. The two eNBs are cooperating in so-called inter eNB CA (Carrier Aggregation) case and connected via asymmetrical IP network. We solve the problem by using broadcasting signals generated in E-UTRAN as synchronization signals. The results show that the time synchronization with the proposed method is possible with the error significantly less than 1 ms which is sufficient considering the time transmission interval is 1 ms in E-UTRAN. This makes this method (with low complexity) more suitable than Network Time Protocol (NTP) in the mobile applications with generated broadcasting signals where time synchronization in asymmetrical network is required.

Keywords: IP scheduled throughput, E-UTRAN, Evolved Universal Terrestrial Radio Access Network, NTP, Network Time Protocol, assymetric network, delay

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979 The Use of Beneficial Microorganisms from Diverse Environments for the Management of Aflatoxin in Maize

Authors: Mathias Twizeyimana, Urmila Adhikari, Julius P. Sserumaga, David Ingham

Abstract:

The management of aflatoxins (naturally occurring toxins produced by certain fungi, most importantly Aspergillus flavus and A. parasiticus) relies mostly on the use of best cultural practices and, in some cases, the use of the biological control consisting of atoxigenic strains inhibiting the toxigenic strains through competition resulting in considerable toxin reduction. At AgBiome, we have built a core collection of over 100,000 fully sequenced microbes from diverse environments and employ both the microbes and their sequences in the discovery of new biological products for disease and pest control. The most common approach to finding beneficial microbes consists of isolating microorganisms from samples collected from diverse environments, selecting antagonistic strains through empirical screening, studying modes of action, and stabilization through the formulation of selected microbial isolates. A total of 608 diverse bacterial strains were screened using a high-throughput assay (48-well assay) to identify strains that inhibit toxigenic A. flavus growth on maize kernels. Active strains in 48-well assay had their pathogen inhibiting activity confirmed using the Flask Assay and were concurrently tested for their ability to reduce the aflatoxin content in maize grains. Strains with best growth inhibition and reduction of aflatoxin were tested in the greenhouse and field trials. From the field trials, three bacterial strains, AFS000009 (Pseudomonas chlororaphis), AFS032321 (Bacillus subtilis), AFS024683 (Bacillus velezensis), had aflatoxin concentrations (ppb) values that were significantly lower than those of inoculated control. The identification of biological products with high efficacy in inhibiting pathogen growth and eventually reducing the aflatoxin content will provide a valuable alternative to control strategies used in aflatoxin contamination management.

Keywords: aflatoxin, microorganism bacteria, biocontrol, beneficial microbes

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978 The Response of Soil Biodiversity to Agriculture Practice in Rhizosphere

Authors: Yan Wang, Guowei Chen, Gang Wang

Abstract:

Soil microbial diversity is one of the important parameters to assess the soil fertility and soil health, even stability of the ecosystem. In this paper, we aim to reveal the soil microbial difference in rhizosphere and root zone, even to pick the special biomarkers influenced by the long term tillage practices, which included four treatments of no-tillage, ridge tillage, continuous cropping with corn and crop rotation with corn and soybean. Here, high-throughput sequencing was performed to investigate the difference of bacteria in rhizosphere and root zone. The results showed a very significant difference of species richness between rhizosphere and root zone soil at the same crop rotation system (p < 0.01), and also significant difference of species richness was found between continuous cropping with corn and corn-soybean rotation treatment in the rhizosphere statement, no-tillage and ridge tillage in root zone soils. Implied by further beta diversity analysis, both tillage methods and crop rotation systems influence the soil microbial diversity and community structure in varying degree. The composition and community structure of microbes in rhizosphere and root zone soils were clustered distinctly by the beta diversity (p < 0.05). Linear discriminant analysis coupled with effect size (LEfSe) analysis of total taxa in rhizosphere picked more than 100 bacterial taxa, which were significantly more abundant than that in root zone soils, whereas the number of biomarkers was lower between the continuous cropping with corn and crop rotation treatment, the same pattern was found at no-tillage and ridge tillage treatment. Bacterial communities were greatly influenced by main environmental factors in large scale, which is the result of biological adaptation and acclimation, hence it is beneficial for optimizing agricultural practices.

Keywords: tillage methods, biomarker, biodiversity, rhizosphere

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977 Carboxyfullerene-Modified Titanium Dioxide Nanoparticles in Singlet Oxygen and Hydroxyl Radicals Scavenging Activity

Authors: Kai-Cheng Yang, Yen-Ling Chen, Er-Chieh Cho, Kuen-Chan Lee

Abstract:

Titanium dioxide nanomaterials offer superior protection for human skin against the full spectrum of ultraviolet light. However, some literature reviews indicated that it might be associated with adverse effects such as cytotoxicity or reactive oxygen species (ROS) due to their nanoscale. The surface of fullerene is covered with π electrons constituting aromatic structures, which can effectively scavenge large amount of radicals. Unfortunately, fullerenes are poor solubility in water, severe aggregation, and toxicity in biological applications when dispersed in solvent have imposed the limitations to the use of fullerenes. Carboxyfullerene acts as the scavenger of radicals for several years. Some reports indicate that carboxyfullerene not only decrease the concentration of free radicals in ambience but also prevent cells from reducing the number or apoptosis under UV irradiation. The aim of this study is to decorate fullerene –C70-carboxylic acid (C70-COOH) on the surface of titanium dioxide nanoparticles (P25) for the purpose of scavenging ROS during the irradiation. The modified material is prepared through the esterification of C70-COOH with P25 (P25/C70-COOH). The binding edge and structure are studied by using Transmission electron microscope (TEM) and Fourier transform infrared (FTIR). The diameter of P25 is about 30 nm and C70-COOH is found to be conjugated on the edge of P25 in aggregation morphology with the size of ca. 100 nm. In the next step, the FTIR was used to confirm the binding structure between P25 and C70-COOH. There are two new peaks are shown at 1427 and 1720 cm-1 for P25/C70-COOH, resulting from the C–C stretch and C=O stretch formed during esterification with dilute sulfuric acid. The IR results further confirm the chemically bonded interaction between C70-COOH and P25. In order to provide the evidence of scavenging radical ability of P25/C70-COOH, we chose pyridoxine (Vit.B6) and terephthalic acid (TA) to react with singlet oxygen and hydroxyl radicals. We utilized these chemicals to observe the radicals scavenging statement via detecting the intensity of ultraviolet adsorption or fluorescence emission. The UV spectra are measured by using different concentration of C70-COOH modified P25 with 1mM pyridoxine under UV irradiation for various duration times. The results revealed that the concentration of pyridoxine was increased when cooperating with P25/C70-COOH after three hours as compared with control (only P25). It indicates fewer radicals could be reacted with pyridoxine because of the absorption via P25/C70-COOH. The fluorescence spectra are observed by measuring P25/C70-COOH with 1mM terephthalic acid under UV irradiation for various duration times. The fluorescence intensity of TAOH was decreased in ten minutes when cooperating with P25/C70-COOH. Here, it was found that the fluorescence intensity was increased after thirty minutes, which could be attributed to the saturation of C70-COOH in the absorption of radicals. However, the results showed that the modified P25/C70-COOH could reduce the radicals in the environment. Therefore, we expect that P25/C70-COOH is a potential materials in using for antioxidant.

Keywords: titanium dioxide, fullerene, radical scavenging activity, antioxidant

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976 The Impact of Oxytetracycline on the Aquaponic System, Biofilter, and Plants

Authors: Hassan Alhoujeiri, Angele Matrat, Sandra Beaufort, Claire joaniss Cassan, Jerome Silvester

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Aquaponics is a sustainable food production technology, and its transition to industrial-scale systems has created several challenges that require further investigation in order to make it a robust process. One of the critical concerns is the potential accumulation of compounds from veterinary treatments, phytosanitary agents, fish feed, or simply from contaminated water sources. The accumulation of these substances could negatively impact fish health, microbial biofilters, and plant growth, thereby disrupting the system’s overall balance and functionality. The lack of legislation and knowledge regarding the presence of such compounds in aquaponic systems raises concerns about their potential impact on both system balance and food safety. In this study, we focused on the effects of oxytetracycline (OTC), an antibiotic commonly used in aquaculture, on both the microbial biofilter and plant growth. Although OTC is rarely applied in aquaponics today, the fish compartment may need to be isolated from the system during treatment, as it inhibits specific bacterial populations, which could affect the microbial biofilter's efficiency. However, questions remain about the aquaponic system's tolerance threshold, particularly in cases of treatment or residual OTC traces post-treatment. This study results indicated a decline in microbial biofilter activity to 20% compared to the control, potentially corresponding to treatments of 41 mg/L of OTC. Analysis of microbial populations in the biofilter, using flow cytometry and microscopy (confocal and scanning electron microscopy), revealed an increase in bacterial mortality without disrupting the microbial biofilm. Additionally, OTC exposure led to noticeable changes in plant morphology (e.g., color) and growth, though it did not fully inhibit development. However, no significant effects were observed on seed germination at the tested concentrations despite a measurable impact on subsequent plant growth.

Keywords: aquaponic, oxytetracycline, nitrifying biofilter, plant, micropollutants, sustainability

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975 Measuring Housing Quality Using Geographic Information System (GIS)

Authors: Silvija ŠIljeg, Ante ŠIljeg, Ivan Marić

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Measuring housing quality is being done on objective and subjective level using different indicators. During the research 5 urban and housing indicators formed according to 58 variables from different housing, domains were used. The aims of the research were to measure housing quality based on GIS approach and to detect critical points of housing in the example of Croatian coastal Town Zadar. The purposes of GIS in the research are to generate models of housing quality indexes by standardisation and aggregation of variables and to examine accuracy model of housing quality index. Analysis of accuracy has been done on the example of variable referring to educational objects availability. By defining weighted coefficients and using different GIS methods high, middle and low housing quality zones were determined. Obtained results can be of use to town planners, spatial planners and town authorities in the process of generating decisions, guidelines, and spatial interventions.

Keywords: housing quality, GIS, housing quality index, indicators, models of housing quality

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974 Characterization of the Lytic Bacteriophage VbɸAB-1 against Drug Resistant Acinetobacter baumannii Isolated from Hospitalized Pressure Ulcers Patients

Authors: M. Doudi, M. H. Pazandeh, L. Rahimzadeh Torabi

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Bedsores are pressure ulcers that occur on the skin or tissue due to being immobile and lying in bed for extended periods. Bedsores have the potential to progress into open ulcers, increasing the possibility of variety of bacterial infection. Acinetobacter baumannii, a pathogen of considerable clinical importance, exhibited a significant correlation with Bedsores (pressure ulcers) infections, thereby manifesting a wide spectrum of antibiotic resistance. The emergence of drug resistance has led researchers to focus on alternative methods, particularly phage therapy, for tackling bacterial infections. Phage therapy has emerged as a novel therapeutic approach to regulate the activity of these agents. The management of bacterial infections greatly benefits from the clinical utilization of bacteriophages as a valuable antimicrobial intervention. The primary objective of this investigation consisted of isolating and discerning potent bacteriophage capable of targeting multi drug-resistant (MDR) and extensively drug-resistant (XDR) bacteria obtained from pressure ulcers. In present study, analyzed and isolated A. baumannii strains obtained from a cohort of patients suffering from pressure ulcers at Taleghani Hospital in Ahvaz, Iran. An approach that included biochemical and molecular identification techniques was used to determine the taxonomic classification of bacterial isolates at the genus and species levels. The molecular identification process was facilitated by using the 16S rRNA gene in combination with universal primers 27 F, and 1492 R. Bacteriophage was obtained through the isolation process conducted on treatment plant sewage located in Isfahan, Iran. The main goal of this study was to evaluate different characteristics of phage, such as their appearance, range of hosts they can infect, how quickly they can enter a host, their stability at varying temperatures and pH levels, their effectiveness in killing bacteria, the growth pattern of a single phage stage, mapping of enzymatic digestion, and identification of proteomics patterns. The findings demonstrated that an examination was conducted on a sample of 50 specimens, wherein 15 instances of A. baumannii were identified. These microorganisms are the predominant Gram-negative agents known to cause wound infections in individuals suffering from bedsores. The study's findings indicated a high prevalence of antibiotic resistance in the strains isolated from pressure ulcers, excluding the clinical strains that exhibited responsiveness to colistin.According to the findings obtained from assessments of host range and morphological characteristics of bacteriophage VbɸAB-1, it can be concluded that this phage possesses specificity towards A. Baumannii BAH_Glau1001 was classified as a member of the Plasmaviridae family. The bacteriophage mentioned earlier showed the strongest antibacterial effect at a temperature of 18 °C and a pH of 6.5. Through the utilization of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis on protein fragments, it was established that the bacteriophage VbɸAB-1 exhibited a size range between 50 and 75 kilodaltons (KDa). The numerous research findings on the effectiveness of phages and the safety studies conducted suggest that the phages studied in this research can be considered as a practical solution and recommended approach for controlling and treating stubborn pathogens in burn wounds among hospitalized patients.

Keywords: acinetobacter baumannii, extremely drug- resistant, phage therapy, surgery wound

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973 Selection of Lead Mobilizing Bacteria from Contaminated Soils and Their Potential in Promoting Plant Growth through Plant Growth Promoting Activity

Authors: Maria Manzoor, Iram Gul, Muhammad Arshad

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Bacterial strains were isolated from contaminated soil collected from Rawalpindi and Islamabad. The strains were investigated for lead resistance and their effect on Pb solubility and PGPR activity. Incubation experiments were carried for inoculated and unoculated soil containing different levels of Pb. Results revealed that few stains (BTM-4, BTM-11, BTM-14) were able to tolerate Pb up to 600 mg L-1, whereas five strains (BTM-3, BTM-6, BTM-10, BTM-21 and BTM-24) showed significant increase in solubility of Pb when compared to all other strains and control. The CaCl2 extractable Pb was increased by 13.6, 6.8, 4.4 and 2.4 folds compared to un-inoculated control soil at increased soil Pb concentration (500, 1000, 1500 and 200 mg kg-1, respectively). The selected bacterial strains (11) were further investigated for plant growth promotion activity through PGPR assays including. Germination and root elongation assays were also conducted under elevated metal concentration in controlled conditions to elucidate the effects of microbial strains upon plant growth and development. The results showed that all the strains tested in this study, produced significantly varying concentrations of IAA, siderophores and gibberellic acid along with ability to phosphorus solubilization index (PSI). The results of germination and root elongation assay further confirmed the beneficial role of the microbial strains in elevating metal stress through PGPR activity. Among all tested strains, BTM-10 significantly improved plant growth. 1.3 and 2.7 folds increase in root and shoot length was observed when compared to control. Which may be attributed to presence of important plant growth promoting enzymes (IAA 74.6 μg/ml; GA 19.23 μg/ml; Sidrophore units 49% and PSI 1.3 cm). The outcome of this study indicates that these Pb tolerant and solubilizing strains may have the potential for plant growth promotion under metal stress and can be used as mediator when coupled with heavy metal hyperaccumulator plants for phytoremediation of Pb contaminated soil.

Keywords: Pb resistant bacteria, Pb mobilizing bacteria, Phytoextraction of Pb, PGPR activity of bacteria

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972 Biodegradation of 2,4-Dichlorophenol by Pseudomonas chlororaphis Strain Isolated from Activated Sludge Sample from a Wastewater Treatment Plant in Durban, South Africa

Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

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Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

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971 Assessment of Bioaerosol and Microbial Volatile Organic Compounds in Different Sections of Library

Authors: Himanshu Lal, Bipasha Ghosh, Arun Srivastava

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A pilot study of indoor air quality in terms of bioaerosol (fungus and bacteria) and few selective microbial volatile organic compounds (MVOCs) was carried out in different indoor sections of a library for two seasons, namely monsoon and post monsoon. Bioaerosol sampling was carried out using Anderson six stage viable sampler at a flow rate of 28.3 L/min while MVOCs were collected on activated charcoal tubes ORBOTM 90 Carboxen 564.Collected MVOCs were desorbed using carbon disulphide (CS2) and analysed by GC-FID. Microscopic identification for fungus was only carried out. Surface dust was collected by sterilised buds and cultured to identify fungal contaminants. Unlike bacterial size distribution, fungal bioaerosol concentration was found to be highest in the fourth stage in different sections of the library. In post monsoon season both fungal bioaerosol (710 to 3292cfu/m3) and bacterial bioaerosol (298 to 1475cfu/m3) were fund at much greater concentration than in monsoon. In monsoon season unlike post monsoon, I/O ratio for both the bioaerosol fractions was more than one. Rain washout could be the reason of lower outdoor concentration in monsoon season. On the contrary most of the MVOCs namely 1-hexene, 1-pentanol and 1-octen-3-ol were found in the monsoon season instead of post monsoon season with the highest being 1-hexene with 7.09µg/m3 concentration. Among the six identified fungal bioaerosol Aspergillus, Cladosporium and Penicillium were found in maximum concentration while Aspergillus niger, Curvuleria lunata, Cladosporium cladosporioides and Penicillium sp., was indentified in surface dust samples. According to regression analysis apart from environmental factors other factors also played an important role. Thus apart from outdoor infiltration and human sources, accumulated surface dust mostly on organic materials like books, wooden furniture and racks can be attributed to being one of the major sources of both fungal bioaerosols as well as MVOCs found in the library.

Keywords: bacteria, Fungi, indoor air, MVOCs

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970 Attention Multiple Instance Learning for Cancer Tissue Classification in Digital Histopathology Images

Authors: Afaf Alharbi, Qianni Zhang

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The identification of malignant tissue in histopathological slides holds significant importance in both clinical settings and pathology research. This paper introduces a methodology aimed at automatically categorizing cancerous tissue through the utilization of a multiple-instance learning framework. This framework is specifically developed to acquire knowledge of the Bernoulli distribution of the bag label probability by employing neural networks. Furthermore, we put forward a neural network based permutation-invariant aggregation operator, equivalent to attention mechanisms, which is applied to the multi-instance learning network. Through empirical evaluation of an openly available colon cancer histopathology dataset, we provide evidence that our approach surpasses various conventional deep learning methods.

Keywords: attention multiple instance learning, MIL and transfer learning, histopathological slides, cancer tissue classification

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969 Profiling of Bacterial Communities Present in Feces, Milk, and Blood of Lactating Cows Using 16S rRNA Metagenomic Sequencing

Authors: Khethiwe Mtshali, Zamantungwa T. H. Khumalo, Stanford Kwenda, Ismail Arshad, Oriel M. M. Thekisoe

Abstract:

Ecologically, the gut, mammary glands and bloodstream consist of distinct microbial communities of commensals, mutualists and pathogens, forming a complex ecosystem of niches. The by-products derived from these body sites i.e. faeces, milk and blood, respectively, have many uses in rural communities where they aid in the facilitation of day-to-day household activities and occasional rituals. Thus, although livestock rearing plays a vital role in the sustenance of the livelihoods of rural communities, it may serve as a potent reservoir of different pathogenic organisms that could have devastating health and economic implications. This study aimed to simultaneously explore the microbial profiles of corresponding faecal, milk and blood samples from lactating cows using 16S rRNA metagenomic sequencing. Bacterial communities were inferred through the Divisive Amplicon Denoising Algorithm 2 (DADA2) pipeline coupled with SILVA database v138. All downstream analyses were performed in R v3.6.1. Alpha-diversity metrics showed significant differences between faeces and blood, faeces and milk, but did not vary significantly between blood and milk (Kruskal-Wallis, P < 0.05). Beta-diversity metrics on Principal Coordinate Analysis (PCoA) and Non-Metric Dimensional Scaling (NMDS) clustered samples by type, suggesting that microbial communities of the studied niches are significantly different (PERMANOVA, P < 0.05). A number of taxa were significantly differentially abundant (DA) between groups based on the Wald test implemented in the DESeq2 package (Padj < 0.01). The majority of the DA taxa were significantly enriched in faeces than in milk and blood, except for the genus Anaplasma, which was significantly enriched in blood and was, in turn, the most abundant taxon overall. A total of 30 phyla, 74 classes, 156 orders, 243 families and 408 genera were obtained from the overall analysis. The most abundant phyla obtained between the three body sites were Firmicutes, Bacteroidota, and Proteobacteria. A total of 58 genus-level taxa were simultaneously detected between the sample groups, while bacterial signatures of at least 8 of these occurred concurrently in corresponding faeces, milk and blood samples from the same group of animals constituting a pool. The important taxa identified in this study could be categorized into four potentially pathogenic clusters: i) arthropod-borne; ii) food-borne and zoonotic; iii) mastitogenic and; iv) metritic and abortigenic. This study provides insight into the microbial composition of bovine faeces, milk, and blood and its extent of overlapping. It further highlights the potential risk of disease occurrence and transmission between the animals and the inhabitants of the sampled rural community, pertaining to their unsanitary practices associated with the use of cattle by-products.

Keywords: microbial profiling, 16S rRNA, NGS, feces, milk, blood, lactating cows, small-scale farmers

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968 Bronchoscopy and Genexpert in the Diagnosis of Pulmonary Tuberculosis in the Indian Private Health Sector: A Short Case Series

Authors: J. J. Mathew

Abstract:

Pulmonary tuberculosis is highly prevalent in the Indian subcontinent. Most cases of pulmonary tuberculosis are diagnosed with sputum examinations and the vast majority of these are undertaken by the government run establishments. However, mycobacterial cultures are not routinely done, unless drug resistance is detected based on clinical response. Modern diagnostic tests like bronchoscopy and Genexpert are not routinely employed in the government institutions for the diagnosis of pulmonary tuberculosis, but have been accepted widely by good private institutions. The utility of these investigations in the private sector is not yet well recognized. This retrospective study aims to assess the usefulness of bronchoscopy and Genexpert in the diagnosis of pulmonary tuberculosis in quaternary care private hospital in India. 30 patients with respiratory symptoms raising the possibility of tuberculosis based on clinical and radiological features, but without any significant sputum production, were subject to bronchoscopy and BAL samples taken for microbiological studies, including Genexpert. 6 out of the 30 patients were found to be Genexpert positive and none of them showed Rifampicin resistance. All the 6 cases had upper zone predominant disease. One of the 6 cases of tuberculosis had another co-existent bacterial infection according to the routine culture studies. 6 other cases were proven to be due to other bacterial infections alone, 2 had a malignant diagnosis and the remaining cases were thought to be non-infective pathologies. The Genexpert results were made available within 48 hours in the 6 positive cases. All of them were commenced on standard anti-tuberculous regimen with excellent clinical response. The other infective cases were also managed successfully based on the drug susceptibilities. The study has shown the usefulness of these investigations as early intervention enabled diagnosis facilitating treatment and prevention of any clinical deterioration. The study lends support to early bronchoscopy and Genexpert testing in suspected cases of pulmonary tuberculosis without significant sputum production, in a high prevalence country which normally relies on sputum examination for the diagnosis of pulmonary tuberculosis.

Keywords: pulmonary, tuberculosis, bronchoscopy, genexpert

Procedia PDF Downloads 244
967 Reduction of Biofilm Formation in Closed Circuit Cooling Towers

Authors: Irfan Turetgen

Abstract:

Closed-circuit cooling towers are cooling units that operate according to the indirect cooling principle. Unlike the open-loop cooling tower, the filler material includes a closed-loop water-operated heat exchanger. The main purpose of this heat exchanger is to prevent the cooled process water from contacting with the external environment. In order to ensure that the hot water is cooled, the water is cooled by the air flow and the circulation water of the tower as it passes through the pipe. They are now more commonly used than open loop cooling towers that provide cooling with plastic filling material. As with all surfaces in contact with water, there is a biofilm formation on the outer surface of the pipe. Although biofilm has been studied very well on plastic surfaces in open loop cooling towers, studies on biofilm layer formed on the heat exchangers of the closed circuit tower have not been found. In the recent study, natural biofilm formation was observed on the heat exchangers of the closed loop tower for 6 months. At the same time, nano-silica coating, which is known to reduce the formation of the biofilm layer, a comparison was made between the two different surfaces in terms of biofilm formation potential. Test surfaces were placed into biofilm reactor along with the untreated control coupons up to 6-months period for biofilm maturation. Natural bacterial communities were monitored to analyze the impact to mimic the real-life conditions. Surfaces were monthly analyzed in situ for their microbial load using epifluorescence microscopy. Wettability is known to play a key role in biofilm formation on surfaces, because characteristics of surface properties affect the bacterial adhesion. Results showed that surface-conditioning with nano-silica significantly reduce (up to 90%) biofilm formation. Easy coating process is a facile and low-cost method to prepare hydrophobic surface without any kinds of expensive compounds or methods.

Keywords: biofilms, cooling towers, fill material, nano silica

Procedia PDF Downloads 127
966 Application of Adaptive Neural Network Algorithms for Determination of Salt Composition of Waters Using Laser Spectroscopy

Authors: Tatiana A. Dolenko, Sergey A. Burikov, Alexander O. Efitorov, Sergey A. Dolenko

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In this study, a comparative analysis of the approaches associated with the use of neural network algorithms for effective solution of a complex inverse problem – the problem of identifying and determining the individual concentrations of inorganic salts in multicomponent aqueous solutions by the spectra of Raman scattering of light – is performed. It is shown that application of artificial neural networks provides the average accuracy of determination of concentration of each salt no worse than 0.025 M. The results of comparative analysis of input data compression methods are presented. It is demonstrated that use of uniform aggregation of input features allows decreasing the error of determination of individual concentrations of components by 16-18% on the average.

Keywords: inverse problems, multi-component solutions, neural networks, Raman spectroscopy

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965 Detection and Identification of Antibiotic Resistant UPEC Using FTIR-Microscopy and Advanced Multivariate Analysis

Authors: Uraib Sharaha, Ahmad Salman, Eladio Rodriguez-Diaz, Elad Shufan, Klaris Riesenberg, Irving J. Bigio, Mahmoud Huleihel

Abstract:

Antimicrobial drugs have played an indispensable role in controlling illness and death associated with infectious diseases in animals and humans. However, the increasing resistance of bacteria to a broad spectrum of commonly used antibiotics has become a global healthcare problem. Many antibiotics had lost their effectiveness since the beginning of the antibiotic era because many bacteria have adapted defenses against these antibiotics. Rapid determination of antimicrobial susceptibility of a clinical isolate is often crucial for the optimal antimicrobial therapy of infected patients and in many cases can save lives. The conventional methods for susceptibility testing require the isolation of the pathogen from a clinical specimen by culturing on the appropriate media (this culturing stage lasts 24 h-first culturing). Then, chosen colonies are grown on media containing antibiotic(s), using micro-diffusion discs (second culturing time is also 24 h) in order to determine its bacterial susceptibility. Other methods, genotyping methods, E-test and automated methods were also developed for testing antimicrobial susceptibility. Most of these methods are expensive and time-consuming. Fourier transform infrared (FTIR) microscopy is rapid, safe, effective and low cost method that was widely and successfully used in different studies for the identification of various biological samples including bacteria; nonetheless, its true potential in routine clinical diagnosis has not yet been established. The new modern infrared (IR) spectrometers with high spectral resolution enable measuring unprecedented biochemical information from cells at the molecular level. Moreover, the development of new bioinformatics analyses combined with IR spectroscopy becomes a powerful technique, which enables the detection of structural changes associated with resistivity. The main goal of this study is to evaluate the potential of the FTIR microscopy in tandem with machine learning algorithms for rapid and reliable identification of bacterial susceptibility to antibiotics in time span of few minutes. The UTI E.coli bacterial samples, which were identified at the species level by MALDI-TOF and examined for their susceptibility by the routine assay (micro-diffusion discs), are obtained from the bacteriology laboratories in Soroka University Medical Center (SUMC). These samples were examined by FTIR microscopy and analyzed by advanced statistical methods. Our results, based on 700 E.coli samples, were promising and showed that by using infrared spectroscopic technique together with multivariate analysis, it is possible to classify the tested bacteria into sensitive and resistant with success rate higher than 90% for eight different antibiotics. Based on these preliminary results, it is worthwhile to continue developing the FTIR microscopy technique as a rapid and reliable method for identification antibiotic susceptibility.

Keywords: antibiotics, E.coli, FTIR, multivariate analysis, susceptibility, UTI

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964 The Lytic Bacteriophage VbɸAB-1 Against Drug-Resistant Acinetobacter Baumannii Isolated from Hospitalized Pressure Ulcers Patients

Authors: M. Doudi, M. H. Pazandeh, L. Rahimzadeh Torabi

Abstract:

Bedsores are pressure ulcers that occur on the skin or tissue due to being immobile and lying in bed for extended periods. Bedsores have the potential to progress into open ulcers, increasing the possibility of a variety of bacterial infections. Acinetobacter baumannii, a pathogen of considerable clinical importance, exhibited a significant correlation with Bedsores (pressure ulcers) infections, thereby manifesting a wide spectrum of antibiotic resistance. The emergence of drug resistance has led researchers to focus on alternative methods, particularly phage therapy, for tackling bacterial infections. Phage therapy has emerged as a novel therapeutic approach to regulate the activity of these agents. The management of bacterial infections greatly benefits from the clinical utilization of bacteriophages as a valuable antimicrobial intervention. The primary objective of this investigation consisted of isolating and discerning potent bacteriophage capable of targeting multi-drug-resistant (MDR) and extensively drug-resistant (XDR) bacteria obtained from pressure ulcers. The present study analyzed and isolated A. baumannii strains obtained from a cohort of patients suffering from pressure ulcers at Taleghani Hospital in Ahvaz, Iran. An approach that included biochemical and molecular identification techniques was used to determine the taxonomic classification of bacterial isolates at the genus and species levels. The molecular identification process was facilitated by using the 16S rRNA gene in combination with universal primers 27 F and 1492 R. Bacteriophage was obtained through the isolation process conducted on treatment plant sewage located in Isfahan, Iran. The main goal of this study was to evaluate different characteristics of phage, such as their appearance, the range of hosts they can infect, how quickly they can enter a host, their stability at varying temperatures and pH levels, their effectiveness in killing bacteria, the growth pattern of a single phage stage, mapping of enzymatic digestion, and identification of proteomics patterns. The findings demonstrated that an examination was conducted on a sample of 50 specimens, wherein 15 instances of A. baumannii were identified. These microorganisms are the predominant Gram-negative agents known to cause wound infections in individuals suffering from bedsores. The study's findings indicated a high prevalence of antibiotic resistance in the strains isolated from pressure ulcers, excluding the clinical strains that exhibited responsiveness to colistin. According to the findings obtained from assessments of host range and morphological characteristics of bacteriophage VbɸAB-1, it can be concluded that this phage possesses specificity towards A. Baumannii BAH_Glau1001 was classified as a member of the Podoviridae family. The bacteriophage mentioned earlier showed the strongest antibacterial effect at a temperature of 18 °C and a pH of 6.5. Through the utilization of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis on protein fragments, it was established that the bacteriophage VbɸAB-1 exhibited a size range between 50 and 75 kilodaltons (KDa). The numerous research findings on the effectiveness of phages and the safety studies conducted suggest that the phages studied in this research can be considered as a practical solution and recommended approach for controlling and treating stubborn pathogens in burn wounds among hospitalized patients. The findings of our research indicated that isolated phages could be an effective antimicrobial and an appreciate candidate for prophylaxis against pressure ulcers.

Keywords: acinetobacter baumannii, extremely drug-resistant, phage therapy, surgery wound

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963 Identification of the Antimicrobial Property of Double Metal Oxide/Bioactive Glass Nanocomposite Against Multi Drug Resistant Staphylococcus aureus Causing Implant Infections

Authors: M. H. Pazandeh, M. Doudi, S. Barahimi, L. Rahimzadeh Torabi

Abstract:

The use of antibiotics is essential in reducing the occurrence of adverse effects and inhibiting the emergence of antibiotic resistance in microbial populations. The necessity for a novel methodology concerning local administration of antibiotics has arisen, with particular focus on dealing with localized infections prompted by bacterial colonization of medical devices or implant materials. Bioactive glasses (BG) are extensively employed in the field of regenerative medicine, encompassing a diverse range of materials utilized for drug delivery systems. In the present investigation, various drug carriers for imipenem and tetracycline, namely single systems BG/SnO2, BG/NiO with varying proportions of metal oxide, and nanocomposite BG/SnO2/NiO, were synthesized through the sol-gel technique. The antibacterial efficacy of the synthesized samples was assessed through the utilization of the disk diffusion method with the aim of neutralizing Staphylococcus aureus as the bacterial model. The current study involved the examination of the bioactivity of two samples, namely BG10SnO2/10NiO and BG20SnO2, which were chosen based on their heightened bacterial inactivation properties. This evaluation entailed the employment of two techniques: the measurement of the pH of simulated body fluid (SBF) solution and the analysis of the sample tablets through X-ray diffraction (XRD), scanning electron microscopy (SEM), and Fourier transform infrared (FTIR) spectroscopy. The sample tablets were submerged in SBF for varying durations of 7, 14, and 28 days. The bioactivity of the composite bioactive glass sample was assessed through characterization of alterations in its surface morphology, structure, and chemical composition. This evaluation was performed using scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, and X-ray diffraction spectroscopy. Subsequently, the sample was immersed in simulated liquids to simulate its behavior in biological environments. The specific body fat percentage (SBF) was assessed over a 28-day period. The confirmation of the formation of a hydroxyapatite surface layer serves as a distinct indicator of bioactivity. The infusion of antibiotics into the composite bioactive glass specimen was done separately, and then the release kinetics of tetracycline and imipenem were tested in simulated body fluid (SBF). Antimicrobial effectiveness against various bacterial strains have been proven in numerous instances using both melt and sol-gel techniques to create multiple bioactive glass compositions. An elevated concentration of calcium ions within a solution has been observed to cause an increase in the pH level. In aqueous suspensions, bioactive glass particles manifest a significant antimicrobial impact. The composite bioactive glass specimen exhibits a gradual and uninterrupted release, which is highly desirable for a drug delivery system over a span of 72 hours. The reduction in absorption, which signals the loss of a portion of the antibiotic during the loading process from the initial phosphate-buffered saline solution, indicates the successful bonding of the two antibiotics to the surfaces of the bioactive glass samples. The sample denoted as BG/10SnO2/10NiO exhibits a higher loading of particles compared to the sample designated as BG/20SnO2 in the context of bioactive glass. The enriched sample demonstrates a heightened bactericidal impact on the bacteria under investigation while concurrently preserving its antibacterial characteristics. Tailored bioactive glass that incorporates hydroxyapatite, with a regulated and efficient release of drugs targeting bacterial infections, holds promise as a potential framework for bone implant scaffolds following rigorous clinical evaluation, thereby establishing potential future biomedical uses. During the modification process, the introduction of metal oxides into bioactive glass resulted in improved antibacterial characteristics, particularly in the composite bioactive glass sample that displayed the highest level of efficiency.

Keywords: antibacterial, bioactive glasses, implant infections, multi drug resistant

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962 Light-Scattering Characteristics of Ordered Arrays Nobel Metal Nanoparticles

Authors: Yassine Ait-El-Aoud, Michael Okomoto, Andrew M. Luce, Alkim Akyurtlu, Richard M. Osgood III

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Light scattering of metal nanoparticles (NPs) has a unique, and technologically important effect on enhancing light absorption in substrates because most of the light scatters into the substrate near the localized plasmon resonance of the NPs. The optical response, such as the resonant frequency and forward- and backward-scattering, can be tuned to trap light over a certain spectral region by adjusting the nanoparticle material size, shape, aggregation state, Metallic vs. insulating state, as well as local environmental conditions. In this work, we examined the light scattering characteristics of ordered arrays of metal nanoparticles and the light trapping, in order to enhance absorption, by measuring the forward- and backward-scattering using a UV/VIS/NIR spectrophotometer. Samples were fabricated using the popular self-assembly process method: dip coating, combined with nanosphere lithography.

Keywords: dip coating, light-scattering, metal nanoparticles, nanosphere lithography

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961 Bacterial Causes of Cerebral Abscess and Impact on Long Term Patient Outcomes

Authors: Umar Rehman, Holly Roy, K. T. Tsang, D. S. Jeyaretna, W Singleton, B. Fisher, P. A. Glew, J. Greig, Peter C. Whitfield

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Introduction: A brain abscess is a life-threatening condition, carrying significant mortality. It requires rapid identification and treatment. Management involves a combination of antibiotics and surgery. The aim of the current study was to identify common bacteria responsible for cerebral abscesses as well as the long term functional and neurological outcomes of patients following treatment in a retrospective series at a single UK neurosurgical centre. Methodology: We analysed patients that had received a diagnosis of 'cerebral abscess' or 'subdural empyema' between June 2002 and June 2018. This was done in the form of a retrospective review. The search resulted in a total of 180 patients; with 37 patients being excluded (spinal abscess, below 18 or non-abscess related admissions). Data were collected from medical case notes including information about demographics, comorbidities, immunosuppression, presentation, size/location of lesions, pathogens, treatment, and outcomes. Results: In total, we analysed 143 patients between the ages of 18-90. Focal neurological deficit and headaches were seen in 84% and 68% of patients respectively. 108 positive brain cultures were seen; with the largest proportion, 59.2% being gram-positive cocci, with strep intermedius being the most common pathogen identified in 13.9% of patients. Of the patients with positive blood cultures (n=11), 72.7% showed the same organism both in the blood and on the brain cultures. Long term outcomes (n=72) revealed that 48% of patients seizure-free without requiring anti-epileptics, 51.3% of patients had full recovery of their neurological symptoms. There was a mortality rate of 13.9% in the series. Conclusion: In conclusion, the largest bacterial cause of abscess within our population was due to gram-positive cocci. The majority of the patient demonstrated full neurological recovery with close to half of patients not requiring anti-epileptics following discharge.

Keywords: bacteria, cerebral abscess, long term outcome, neurological deficit

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960 Antimicrobial Action and Its Underlying Mechanism by Methanolic Seed Extract of Syzygium cumini on Bacillus subtilis

Authors: Alok Kumar Yadav, Saurabh Saraswat, Preeti Sirohi, Manjoo Rani, Sameer Srivastava, Manish Pratap Singh, Nand K. Singh

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The development of antibiotic resistance in bacteria is increasing at an alarming rate, and this is considered as one of the most serious threats in the history of medicine, and an alternative solution should be derived so as to tackle this problem. In many countries, people use the medicinal plants for the treatment of various diseases as these are cheaper, easily available and least toxic. Syzygium cumini is used for the treatment of various kinds of diseases but their mechanism of action is not reported. The antimicrobial activity of Syzygium cumini was tested by the well diffusion assay and zone of inhibition was reported to be 20.06 mm as compared to control with MIC of 0.3 mg/ml. Genomic DNA fragmentation of Bacillus subtilis revealed apoptosis and FE-SEM indicate cell wall cracking on several intervals of time. Propidium iodide staining results showed that few bacterial cells were stained in the control and population of stained cells increase after exposing them for various period of time. Flow cytometric kinetic data analysis on the membrane permeabilization in bacterial cell showed the significant contribution of antimicrobial potential of the seed extract on antimicrobial-induced permeabilization. Two components of Syzygium cumini methanolic seed extract was found to be quite active against four enzymes like PDB ID- 1W5D, 4OX3, 3MFD and 5E2F which have a very crucial role in membrane synthesis in Bacillus subtilis by in silico analysis. Through in silico analysis, lupeol showed highest binding energy for macromolecule 1W5D and 4OX3 whereas stigmasterol showed the highest binding energy for macromolecule 3MFD and 5E2F respectively. It showed that methanolic seed extract of Syzygium cumini can be used for the inhibition of foodborne infections caused by Bacillus subtilis and also as an alternative of prevalent antibiotics.

Keywords: antibiotics, Bacillus subtilis, inhibition, Syzygium cumini

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959 Effects of Indole on Aerobic Biodegradation of Butanoic Acid by Pseudomonas aeruginosa and Serratia marcescens

Authors: J. B. J. Njalam’mano, E. M. N. Chirwa

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In low resource settings in Africa and other developing regions, pit latrines remain the dominant basic minimum acceptable form of sanitation. However, unpleasant smells-malodours emitted from faecal sludge in the pit latrines, which elicit disgusting or repulsive response, are one of the factors that thwart people to use latrines and instead opt for open defecation as an alternative. This provides an important but often overlooked major impediment, dissuading people from adopting and using the pit latrines hence affecting successful, effective sanitation promotion. The malodours are primarily attributed to four odorants: butanoic acid (C₄H₈O₂), dimethyl trisulphide (C₂H₆S₃), indole (C₈H₇N) and para-cresol (C₇H₈O). Several pit latrine deodorisation methods such as addition of carbonous materials, use of ventilation systems and urine separation are available, and they continue to occupy their niche, but social, economic, environmental and technological shortfalls remain. Bioremediation has been gaining popularity because it is inexpensive, simple to operate and environmentally friendly. Recently, the biodegradation of butanoic acid as individual odorant has been studied. However, to the best of our knowledge, there have been no kinetic studies of the butanoic acid in the presence of other key odorous compounds. In this study, a series of experiments were conducted to investigate the effects of indole on the removal of butanoic acid under aerobic conditions using indigenous bacteria strains, Pseudomonas aeruginosa, and Serratia marcescens isolated from faecal sludge as pure cultures as well as mixed cultures. In this purpose, butanoic acid removal was performed in a batch reactor containing the bacterial strains in mineral salt medium (MSM) amended with 3000 ppm of butanoic acid at the temperature of 30°C, under continuous stirring rate of 150 rpm and the concentration of indole was varied from 50-200 ppm. The initial pH of the solution was in the range of 6.0-7.2. Overall, there were significant differences in the bacterial growth rate and total butanoic acid removal dependent on the concentration of indole in the solution.

Keywords: biodegradation, butanoic acid, indole, pit latrine

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958 Microbial Biogeography of Greek Olive Varieties Assessed by Amplicon-Based Metagenomics Analysis

Authors: Lena Payati, Maria Kazou, Effie Tsakalidou

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Table olives are one of the most popular fermented vegetables worldwide, which along with olive oil, have a crucial role in the world economy. They are highly appreciated by the consumers for their characteristic taste and pleasant aromas, while several health and nutritional benefits have been reported as well. Until recently, microbial biogeography, i.e., the study of microbial diversity over time and space, has been mainly associated with wine. However, nowadays, the term 'terroir' has been extended to other crops and food products so as to link the geographical origin and environmental conditions to quality aspects of fermented foods. Taking the above into consideration, the present study focuses on the microbial fingerprinting of the most important olive varieties of Greece with the state-of-the-art amplicon-based metagenomics analysis. Towards this, in 2019, 61 samples from 38 different olive varieties were collected at the final stage of ripening from 13 well spread geographical regions in Greece. For the metagenomics analysis, total DNA was extracted from the olive samples, and the 16S rRNA gene and ITS DNA region were sequenced and analyzed using bioinformatics tools for the identification of bacterial and yeasts/fungal diversity, respectively. Furthermore, principal component analysis (PCA) was also performed for data clustering based on the average microbial composition of all samples from each region of origin. According to the composition, results obtained, when samples were analyzed separately, the majority of both bacteria (such as Pantoea, Enterobacter, Roserbergiella, and Pseudomonas) and yeasts/fungi (such as Aureobasidium, Debaromyces, Candida, and Cladosporium) genera identified were found in all 61 samples. Even though interesting differences were observed at the relative abundance level of the identified genera, the bacterial genus Pantoea and the yeast/fungi genus Aureobasidium were the dominant ones in 35 and 40 samples, respectively. Of note, olive samples collected from the same region had similar fingerprint (genera identified and relative abundance level) regardless of the variety, indicating a potential association between the relative abundance of certain taxa and the geographical region. When samples were grouped by region of origin, distinct bacterial profiles per region were observed, which was also evident from the PCA analysis. This was not the case for the yeast/fungi profiles since 10 out of the 13 regions were grouped together mainly due to the dominance of the genus Aureobasidium. A second cluster was formed for the islands Crete and Rhodes, both of which are located in the Southeast Aegean Sea. These two regions clustered together mainly due to the identification of the genus Toxicocladosporium in relatively high abundances. Finally, the Agrinio region was separated from the others as it showed a completely different microbial fingerprinting. However, due to the limited number of olive samples from some regions, a subsequent PCA analysis with more samples from these regions is expected to yield in a more clear clustering. The present study is part of a bigger project, the first of its kind in Greece, with the ultimate goal to analyze a larger set of olive samples of different varieties and from different regions in Greece in order to have a reliable olives’ microbial biogeography.

Keywords: amplicon-based metagenomics analysis, bacteria, microbial biogeography, olive microbiota, yeasts/fungi

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957 Impact of Ecosystem Engineers on Soil Structuration in a Restored Floodplain in Switzerland

Authors: Andreas Schomburg, Claire Le Bayon, Claire Guenat, Philip Brunner

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Numerous river restoration projects have been established in Switzerland in recent years after decades of human activity in floodplains. The success of restoration projects in terms of biodiversity and ecosystem functions largely depend on the development of the floodplain soil system. Plants and earthworms as ecosystem engineers are known to be able to build up a stable soil structure by incorporating soil organic matter into the soil matrix that creates water stable soil aggregates. Their engineering efficiency however largely depends on changing soil properties and frequent floods along an evolutive floodplain transect. This study, therefore, aims to quantify the effect of flood frequency and duration as well as of physico-chemical soil parameters on plants’ and earthworms’ engineering efficiency. It is furthermore predicted that these influences may have a different impact on one of the engineers that leads to a varying contribution to aggregate formation within the floodplain transect. Ecosystem engineers were sampled and described in three different floodplain habitats differentiated according to the evolutionary stages of the vegetation ranging from pioneer to forest vegetation in a floodplain restored 15 years ago. In addition, the same analyses were performed in an embanked adjacent pasture as a reference for the pre-restored state. Soil aggregates were collected and analyzed for their organic matter quantity and quality using Rock Eval pyrolysis. Water level and discharge measurements dating back until 2008 were used to quantify the return period of major floods. Our results show an increasing amount of water stable aggregates in soil with increasing distance to the river and show largest values in the reference site. A decreasing flood frequency and the proportion of silt and clay in the soil texture explain these findings according to F values from one way ANOVA of a fitted mixed effect model. Significantly larger amounts of labile organic matter signatures were found in soil aggregates in the forest habitat and in the reference site that indicates a larger contribution of plants to soil aggregation in these habitats compared to the pioneer vegetation zone. Earthworms’ contribution to soil aggregation does not show significant differences in the floodplain transect, but their effect could be identified even in the pioneer vegetation with its large proportion of coarse sand in the soil texture and frequent inundations. These findings indicate that ecosystem engineers seem to be able to create soil aggregates even under unfavorable soil conditions and under frequent floods. A restoration success can therefore be expected even in ecosystems with harsh soil properties and frequent external disturbances.

Keywords: ecosystem engineers, flood frequency, floodplains, river restoration, rock eval pyrolysis, soil organic matter incorporation, soil structuration

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956 Prevalence, Antimicrobial Susceptibility Pattern and Associated Risk Factors for Salmonella Species and Escherichia coli from Raw Meat at Butchery Houses in Mekelle, Tigray, Ethiopia

Authors: Haftay Abraha Tadesse, Atsebaha Gebrekidan Kahsay, Mahumd Abdulkader

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Background: Salmonella species and Escherichia coli are important foodborne pathogens affecting humans and animals. They are among the most important causes of infection that are associated with the consumption of contaminated food. This study was aimed to determine the prevalence, antimicrobial susceptibility patterns and associated risk factors for Salmonella species and E. coli in raw meat from butchery houses of Mekelle, Northern Ethiopia. Methodology: A cross-sectional study was conducted from January to September 2019. Socio-demographic data and risk factors were collected using a predesigned questionnaire. Meat samples were collected aseptically from the butchery houses and transported using icebox to Mekelle University, College of Veterinary Sciences for the isolation and identification of Salmonella species and E. coli, Antimicrobial susceptibility patterns were determined using Kirby disc diffusion method. Data obtained were cleaned and entered into Statistical Package for the Social Sciences version 22 and logistic regression models with odds ratio were calculated. P-value < 0.05 was considered as statistically significant. Results: A total of 153 out of 384 (39.8%) of the meat specimens were found to be contaminated. The contamination of Salmonella species and E. coli were 15.6% (n=60) and 20.8%) (n=80), respectively. Mixed contamination (Salmonella species and E. coli) was observed in 13 (3.4 %) of the analyzed. Poor washing hands regularly (AOR = 8.37; 95% CI: 2.75-25.50) and not using gloves during meat handling (AOR=11. 28; 95% CI: (4.69 27.10) were associated with an overall bacterial contamination.About 95.5% of the tested isolates were sensitive to chloramphenicol and norfloxacin while the resistance of amoxyclav_amoxicillin and erythromycin were both isolated bacteria species. The overall multidrug resistance pattern for Salmonella and E. coli were 51.4% (n=19) and 31.8% (14), respectively. Conclusion: Of the 153 (153/384) contaminated raw meat, 60 (15.6%) and 80 (20.8%) were contaminated by Salmonella species and E. coli, respectively. Poor hand washing practice and not using glove during meat handling showed significant association with bacterial contamination. Multidrug-resistant showed in Salmonella species and E. coli were 19 (51.4%) and 14 (31.8%), respectively.

Keywords: antimicrobial susceptibility test, butchery houses, e. coli, salmonella species

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