Search results for: immobilized bacteria

Authors: Nélio Veiga, Paula Ferreira, Tiago Correia, Maria J. Correia, Carlos Pereira, Odete Amaral, Ilídio J. Correia

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Introduction: The application of fissure sealants is considered to be an important primary prevention method used in dental medicine. However, the formation of microleakage gaps between tooth enamel and the fissure sealant applied is one of the most common reasons of dental caries development in teeth with fissure sealants. The association between various dental biomaterials may limit the major disadvantages and limitations of biomaterials functioning in a complementary manner. The present study consists in the incorporation of a cariostatic agent – silver diamine fluoride (SDF) – in a resin-based fissure sealant followed by the study of release kinetics by spectrophotometry analysis of the association between both biomaterials and assessment of the inhibitory effect on the growth of the reference bacterial strain Streptococcus mutans (S. mutans) in an in vitro study. Materials and Methods: An experimental in vitro study was designed consisting in the entrapment of SDF (Cariestop^® 12% and 30%) into a commercially available fissure sealant (Fissurit^®), by photopolymerization and photocrosslinking. The same sealant, without SDF was used as a negative control. The effect of the sealants on the growth of S. mutans was determined by the presence of bacterial inhibitory halos in the cultures at the end of the incubation period. In order to confirm the absence of bacteria in the surface of the materials, Scanning Electron Microscopy (SEM) characterization was performed. Also, to analyze the release profile of SDF along time, spectrophotometry technique was applied. Results: The obtained results indicate that the association of SDF to a resin-based fissure sealant may be able to increase the inhibition of S. mutans growth. However, no SDF release was noticed during the in vitro release studies and no statistical significant difference was verified when comparing the inhibitory halo sizes obtained for test and control group.  Conclusions: In this study, the entrapment of SDF in the resin-based fissure sealant did not potentiate the antibacterial effect of the fissure sealant or avoid the immediate development of dental caries. The development of more laboratorial research and, afterwards, long-term clinical data are necessary in order to verify if this association between these biomaterials is effective and can be considered for being used in oral health management. Also, other methodologies for associating cariostatic agents and sealant should be addressed.

Keywords: biomaterial, fissure sealant, primary prevention, silver diamine fluoride

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Authors: Jung-En Kuan, Whei-Fen Wu

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In this study, a symbiotic bacteria from yellow mealworm's (Tenebrio molitor) mid-gut was isolated with characteristics of growth on minimal-tributyrin medium. After a PCR-amplification of its 16s rDNA, the resultant nucleotide sequences were then analyzed by schemes of the phylogeny trees. Accordingly, it was designated as Pseudomonas nitroreducens D-01. Next, by searching the lipolytic enzymes in its protein data bank, one of those potential lipolytic α/β hydrolases was identified, again using PCR-amplification and nucleotide-sequencing methods. To construct an expression of this lipolytic gene in plasmids, the target-gene primers were then designed, carrying the C-terminal his-tag sequences. Using the vector pET21a, a recombinant lipolytic hydrolase D gene with his-tag nucleotides was successfully cloned into it, of which the lipolytic D gene is under a control of the T7 promoter. After transformation of the resultant plasmids into Eescherichia coli BL21 (DE3), an IPTG inducer was used for the induction of the recombinant proteins. The protein products were then purified by metal-ion affinity column, and the purified proteins were found capable of forming a clear zone on tributyrin agar plate. Shortly, its enzyme activities were determined by degradation of p-nitrophenyl ester(s), and the substantial yellow end-product, p-nitrophenol, was measured at O.D.405 nm. Specifically, this lipolytic enzyme efficiently targets p-nitrophenyl butyrate. As well, it shows the most reactive activities at 40°C, pH 8 in potassium phosphate buffer. In thermal stability assays, the activities of this enzyme dramatically drop when the temperature is above 50°C. In metal ion assays, MgCl₂ and NH₄Cl induce the enzyme activities while MnSO₄, NiSO₄, CaCl₂, ZnSO₄, CoCl₂, CuSO₄, FeSO₄, and FeCl₃ reduce its activities. Besides, NaCl has no effects on its enzyme activities. Most organic solvents decrease the activities of this enzyme, such as hexane, methanol, ethanol, acetone, isopropanol, chloroform, and ethyl acetate. However, its enzyme activities increase when DMSO exists. All the surfactants like Triton X-100, Tween 80, Tween 20, and Brij35 decrease its lipolytic activities. Using Lineweaver-Burk double reciprocal methods, the function of the enzyme kinetics were determined such as Km = 0.488 (mM), Vmax = 0.0644 (mM/min), and kcat = 3.01x10³ (s⁻¹), as well the total efficiency of kcat/Km is 6.17 x10³ (mM⁻¹/s⁻¹). Afterwards, based on the phylogenetic analyses, this lipolytic protein is classified to type IV lipase by its homologous conserved region in this lipase family.

Keywords: enzyme, esterase, lipotic hydrolase, type IV

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Authors: N. Balaban, A. Buernstein, F. Gelman, Z. Ronen

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Brominated flame retardants are widespread pollutants, and are known to be toxic, carcinogenic, endocrinic disrupting as well as recalcitrant. The industrial complex Neot Hovav, in the Northern Negev, Israel, is situated above a fractured chalk aquitard, which is polluted by a wide variety of halogenated organic compounds. Two of the abundant pollutants found in the site are Dibromoneopentyl-glycol (DBNPG) and tribromoneopentyl-alcohol (TBNPA). Due to the elusive nature of the groundwater flow, it is difficult to connect between the spatial changes in contaminant concentrations to degradation. In this study, we attempt to determine whether these compounds are biodegraded in the groundwater, and to gain a better understanding concerning the bacterial community in the groundwater. This was achieved through the application of compound-specific isotope analysis (CSIA) of carbon (13^C/12^C) and bromine (81^Br/79^Br), and new-generation MiSeq pyrosequencing. The sampled boreholes were distributed among three main areas of the industrial complex: around the production plant of TBNPA and DBNPG; along the Hovav Wadi (small ephemeral stream) which crosses and drains the industrial complex; and downstream to the industrial area. TBNPA and DBNPG are found in all three areas, with no clear connection to the proximity of the borehole to the production plant. Initial isotopic data of TBNPA from boreholes in the area surrounding the production plant, reveal no changes in the carbon and bromine isotopic values. When observing the microbial groundwater community, the dominant phylum is Proteobacteria. Known anaerobic dehalogenating bacteria such as Dehalococcoides from the Chloroflexi phylum have also been detected. A statistical comparison of the groundwater microbial diversity using a multi-variant ordination of non-metric multidimensional scaling (NMDS) reveals three main clusters in accordance to spatial location in the industrial complex: all the boreholes sampled adjacent to the production plant cluster together and separately from the Wadi Hovav boreholes cluster and the downstream to the industrial area borehole cluster. This work provides the basis for the development and implication of an isotopic fractionation based tool for assessing the biodegradation of brominated organic compounds in contaminated environments, and a novel attempt to characterize the spatial microbial diversity in the contaminated site.

Keywords: biodegradation, brominated flame retardants, groundwater, isotopic fractionation, microbial diversity

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Authors: Fatemeh Elmi, Zahra Etemadifar

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Improvement of soil mechanical properties is crucial before its use in construction, as the low mechanical strength and unstable structure of soil in many parts of the world can lead to the destruction of engineering infrastructure, resulting in financial and human losses. Although, conventional methods, such as chemical injection, are often utilized to enhance soil strength and stiffness, they are generally expensive, require heavy machinery, and cause significant environmental effects due to chemical usage, and also disrupt urban infrastructure. Moreover, they are not suitable for treating large volume of soil. Recently, an alternative method to improve various soil properties, including strength, hardness, and permeability, has received much attention: the application of biological methods. One of the most widely used is biocementation, which is based on the microbial precipitation of calcium carbonte crystalls using ureolytic bacteria However, there are still limitations to its large-scale use that need to be resolved before it can be commercialized. These issues have not received enough attention in prior research. One limitation of MICP (microbially induced calcium carbonate precipitation) is that microorganisms cannot operate effectively in harsh and variable environments, unlike the controlled conditions of a laboratory. Another limitation of applying this technique on a large scale is the high cost of producing a substantial amount of bacterial culture and reagents required for soil treatment. Therefore, the purpose of the present study was to investigate soil improvement using the biocementation activity of poly-extremophile, calcium carbonate crystal- producing bacterial strain, Bhargavaea cecembensis N1, in whey as an inexpensive medium. This strain was isolated and molecularly identified from sandy soils in our previous research, and its 16S rRNA gene sequences was deposited in the NCBI Gene Bank with an accession number MK420385. This strain exhibited a high level of urease activity (8.16 U/ml) and produced a large amount of calcium carbonate (4.1 mg/ ml). It was able to improve the soil by increasing the compressive strength up to 205 kPa and reducing permeability by 36%, with 20% of the improvement attributable of calcium carbonate production. This was achieved using this strain in a whey culture medium. This strain can be an eco-friendly and economical alternative to conventional methods in soil stabilization, and other MICP related applications.

Keywords: biocementation, Bhargavaea cecembensis, soil improvement, whey culture medium

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Authors: Raghuram Kandimalla, Sanjeeb Kalita, Bhaswati Choudhury, Jibon Kotoky

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The quest for developing an ideal suture material prompted our interest to develop a novel suture with advantageous characteristics to market available ones. We developed novel suture biomaterial from muga silk (Antheraea assama) and ramie (Boehmeria nivea) plant fiber. Field emission scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDX), attenuated total reflection fourier transform infrared spectroscopy (ATR-FTIR) and thermo gravimetric analysis (TGA) results revealed the physicochemical properties of the fibers which supports the suitability of fibers for suture fabrication. Tensile properties of the prepared sutures were comparable with market available sutures and it found to be biocompatible towards human erythrocytes and nontoxic to mammalian cells. The prepared sutures completely healed the superficial deep wound incisions within seven days in adult male wister rats leaving no rash and scar. Histopathology studies supports the wound healing ability of sutures, as rapid synthesis of collagen, connective tissue and other skin adnexal structures were observed within seven days of surgery. Further muga suture surface modified by exposing the suture to oxygen plasma which resulted in formation of nanotopography on suture surface. Broad spectrum antibiotic amoxicillin was functionalized on the suture surface to prepare an advanced antimicrobial muga suture. Surface hydrophilicity induced by oxygen plasma results in an increase in drug-impregnation efficiency of modified muga suture by 16.7%. In vitro drug release profiles showed continuous and prolonged release of amoxicillin from suture up to 336 hours. The advanced muga suture proves to be effective against growth inhibition of Staphylococcus aureus and Escherichia coli, whereas normal muga suture offers no antibacterial activity against both types of bacteria. In vivo histopathology studies and colony-forming unit count data revealed accelerated wound healing activity of advanced suture over normal one through rapid synthesis and proliferation of collagen, hair follicle and connective tissues.

Keywords: sutures, biomaterials, silk, Ramie

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Authors: Reenu Yadav, Abhay Asthana, S. K. Yadav

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Purpose: The aim of the present work was to develop an oral gel for brushing with an antimicrobial activity which will cure/protect from various periodontal diseases such as periodontitis, gingivitis, and pyorrhea. Methods: Plant materials procured from local suppliers, extracted and standardized. Screening of antimicrobial activity was carried out with the help of disk diffusion method. The gel was formulated by dried extracts of Beautea monosperma and Cordia obliquus. Gels were evaluated on various parameters and standardization of the formulation was performed. The release of drugs was studied in pH 6.8 using a mastication device.Total phenolic and flavonoid contents were estimated by folin-Ciocalteu and aluminium chloride method, and stability studies were performed (40°C and RH 75% ± 5% for 90 days) to assess the effect of temperature and humidity on the concentration of phenolic and flavonoid contents. The results of accelerated stability conditions were compared with that of samples kept at controlled conditions (RT). The control samples were kept at room temperature (25°C, 35% RH for 180 days). Results: Results are encouraging; extracts possess significant antimicrobial activity at very low concentration (15µg/disc, 20µg/disc and 15 µg/ disc) on oral pathogenic bacteria. The formulation has optimal characteristics, as well as has a pleasant appearance, fragrance, texture, and taste, is highly acceptable by the volunteers. The diffusion coefficient values ranged from 0.6655 to 0.9164. Since the R values of korsmayer papas were close to 1, Drug release from formulation follows matrix diffusion kinetics. Hence, diffusion was the mechanism of the drug release. Formulation follows non-Fickian transport mechanism. Most Formulations released 50 % of their contents within 25-30 minutes. Results obtained from the accelerated stability studies are indicative of a slight reduction in flavonoids and phenolic contents with time on long time storage. When measured degradation under ambient conditions, degradation was significantly lower than in accelerated stability study. Conclusion: Plant extracts possess compounds with antimicrobial properties can be used as. Developed formulation will cure/protect from various periodontal diseases. Further development and evaluations oral gel including the isolated compounds on the commercial scale and their clinical and toxicological studies are the future challenges.

Keywords: herbal gel, dental care, ambient conditions, commercial scale

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Authors: Ziani Borhane Eddine Cherif, Hazzi Mohamed, Mouhouche Fazia

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The vegetal extracts of aromatic and medicinal plants start to have much of interest like potential sources of natural bioactive molecules. Many features are conferred by the nature of the chemical function of their major constituents (phenol, alcohol, aldehyde, cetone). This biopotential lets us to focalize on the study of three main biological activities, the antioxidant, antibiotic and insecticidal activities of six Algerian aromatic plants in the aim of making in evidence by the chromatographic analysis (CPG and CG/SM) the phytochemical compounds implicating in this effects. The contents of Oxygenated monoterpenes represented the most prominent group of constituents in the majority of plants. However, the α-Terpineol (28,3%), Carvacrol (47,3%), pulégone (39,5%), Chrysanthenone (27,4%), Thymol 23,9%, γ-Terpinene 23,9% and 2-Undecanone(94%) were the main components. The antioxyding activity of the Essential oils and no-volatils extracts was evaluated in vitro using four tests: inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the 2,2-Azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) radical-scavenging activity (ABTS•+), the thiobarbituric acid reactive substances (TBARS) assays and the reducing power. The measures of the IC50 of these natural compounds revealed potent activity (between 254.64-462.76mg.l-1), almost similar to that of BHT, BHA, Tocopherol and Ascorbic acid (126,4-369,1 mg.l-1) and so far than the Trolox one (IC50= 2,82mg.l-1). Furthermore, three ethanol extracts were found to be remarkably effective toward DPPH and ABTS inhibition, compared to chemical antioxidant BHA and BHT (IC = 9.8±0.1 and 28±0.7 mg.l-1, respectively); for reducing power test it has also exhibited high activity. The study on the insecticidal activity effect by contact, inhalation, fecundity and fertility of Callosobruchus maculatus and Tribolium confusum showed a strong potential biocide reaching 95-100% mortality only after 24 hours. The antibiotic activity of our essential oils were evaluated by a qualitative study (aromatogramme) and quantitative (MIC, MBC and CML) on four bacteria (Gram+ and Gram-) and one strain of pathogenic yeast, the results of these tests showed very interesting action than that induced by the same reference antibiotics (Gentamycin, and Nystatin Ceftatidine) such that the inhibition diameters and MIC values for tested microorganisms were in the range of 23–58 mm and 0.015–0.25%(v/v) respectively.

Keywords: aromatic plants, essential oils, no-volatils extracts, bioactive molecules, antioxidant activity, insecticidal activity, antibiotic activity

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Authors: Katarzyna Miśkiewicz, Magdalena Lasoń-Rydel, Małgorzata Krępska, Katarzyna Sieczyńska, Iwona Masłowska-Lipowicz, Katarzyna Ławińska

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The aim of the study was to analyse a selection of 12 pasteurised products made from fruit and vegetables, such as fruit juices, fruit drinks, jams, marmalades and jam produced by small regional factories as well as large food corporations. The research was carried out as part of the project "Innovative system of healthy and regional food distribution", funded by the Ministry of Education and Science (Poland), which aims to create an economically and organisationally strong agri-food industry in Poland through effective cooperation between scientific and socio-economic actors. The main activities of the project include support for the creation of new distribution channels for regional food products and their easy access to a wide group of potential customers while maintaining the highest quality standards. One of the key areas of the project is food quality analyses conducted to indicate the competitive advantage of regional products. Presented here are studies on the content of sugars, vitamin C, preservatives, synthetic colours, sweeteners, sodium and potassium, as well as studies on the microbiological purity of selected products made from fruit and vegetables. The composition of products made from fruit and vegetables varies greatly and depends on both the type of raw material and the way it is processed. Of the samples tested, fruit drinks contained the least amount of sugars, and jam and marmalade made by large producers and bought in large chain stores contained the most. However, the low sugar content of some fruit drinks is due to the presence of the sweetener sucralose in their composition. The vitamin C content of the samples varied, being higher in products where it was added during production. All products made in small local factories were free of food additives such as preservatives, sweeteners and synthetic colours, indicating their superiority over products made by large producers. Products made in small local factories were characterised by a relatively high potassium content. The microbiological purity of commercial products was confirmed - no Salmonella spp. were detected, and the number of mesophilic bacteria, moulds, yeasts, and β-glucuronidase-positive E. coli was below the limit of quantification.

Keywords: fruit and vegetable products, sugars, food additives, HPLC, ICP-OES

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Authors: Falana Yetunde Olaitan, Ijah U. J. J, Solebo Shakirat O.

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Genetically modified crops are already phenomenally successful and are grown worldwide in more than eighteen countries on more than 67 million hectares. Nigeria, in October 2018, approved Bacillus thuringiensis (Bt) cotton and maize; therefore, the need to carry out environmental risk assessment studies. A total of 15 4L octagonal ceramic pots were filled with 4kg of soil and placed on the bench in 2 rows of 10 pots each and the 3rd row of 5 pots, 1st-row pots were used to plant GM cotton seeds, while the 2nd-row pots were used for non-GM cotton seeds and the 3rd row of 5 pots served as control, all in the screen house. Soil samples for metagenomic DNA extraction were collected at random and at the monthly interval after planting at a distance of 2mm from the plant’s root and at a depth of 10cm using a sterile spatula. Soil samples for physicochemical analysis were collected before planting and after harvesting the GM and non-GM crops as well as from the control soil. The DNA was extracted, quantified and sequenced; Sample 1A (DNA from GM cotton Soil at 1st interval) gave the lowest sequence read with 0.853M while sample 2B (DNA from GM cotton Soil at 2nd interval) gave the highest with 5.785M, others gave between 1.8M and 4.7M. The samples treatment were grouped into four, Group 1 (GM cotton soil from 1 to 3 intervals) had between 800,000 and 5,700,000 strains of microbes (SOM), Group 2 (non GM cotton soil from 1 to 3 intervals) had between 1,400,600 and 4,200,000 SOM, Group 3 (control soil) had between 900,000 and 3,600,000 SOM and Group 4 (initial soil) had between 3,700,000 and 4,000,000 SOM. The microbes observed were predominantly bacteria (including archaea), fungi, dark matter alongside protists and phages. The predominant bacterial groups were the Terrabacteria (Bacillus funiculus, Bacillus sp.), the Proteobacteria (Microvirga massiliensis, sphingomonas sp.) and the Archaea (Nitrososphaera sp.), while the fungi were Aspergillus fischeri and Fusarium falciforme. The comparative analysis between groups was done using JACCARD PERMANOVA beta diversity analysis at P-value not more than 0.76 and there was no significant pair found. The pH for initial, GM cotton, non-GM cotton and control soil were 6.28, 6.26, 7.25, 8.26 and the percentage moisture was 0.63, 0.78, 0.89 and 0.82, respectively, while the percentage Nitrogen was observed to be 17.79, 1.14, 1.10 and 0.56 respectively. Other parameters include, varying concentrations of Potassium (0.46, 1,284.47, 1,785.48, 1,252.83 mg/kg) and Phosphorus (18.76, 17.76, 16.87, 15.23 mg/kg) were recorded for the four treatments respectively. The soil consisted mainly of silt (32.09 to 34.66%) and clay (58.89 to 60.23%), reflecting the soil texture as silty – clay. The results were then tested with ANOVA at less than 0.05 P-value and no pair was found to be significant as well. The results suggest that the GM crops have no significant effect on microbial ecology and physicochemical properties of the soil and, in turn, no direct or indirect effects on human health.

Keywords: genetically modified crop, microbial ecology, physicochemical properties, metagenomics, DNA, soil

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Authors: D. Berdous, H. Ferfera-Harrar

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Superabsorbent polymers (SAPs) or hydrogels with three-dimensional hydrophilic network structure are high-performance water absorbent and retention materials. The in situ synthesis of metal nanoparticles within polymeric network as antibacterial agents for bio-applications is an approach that takes advantage of the existing free-space into networks, which not only acts as a template for nucleation of nanoparticles, but also provides long term stability and reduces their toxicity by delaying their oxidation and release. In this work, SAP/nanosilver nanocomposites were successfully developed by a unique green process at room temperature, which involves in situ formation of silver nanoparticles (AgNPs) within hydrogels as a template. The aim of this study is to investigate whether these AgNPs-loaded hydrogels are potential candidates for antimicrobial applications. Firstly, the superabsorbents were prepared through radical copolymerization via grafting and crosslinking of acrylamide (AAm) onto chitosan backbone (Cs) using potassium persulfate as initiator and N,N’-methylenebisacrylamide as the crosslinker. Then, they were hydrolyzed to achieve superabsorbents with ampholytic properties and uppermost swelling capacity. Lastly, the AgNPs were biosynthesized and entrapped into hydrogels through a simple, eco-friendly and cost-effective method using aqueous silver nitrate as a silver precursor and curcuma longa tuber-powder extracts as both reducing and stabilizing agent. The formed superabsorbents nanocomposites (Cs-g-PAAm)/AgNPs were characterized by X-ray Diffraction (XRD), UV-visible Spectroscopy, Attenuated Total reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Inductively Coupled Plasma (ICP), and Thermogravimetric Analysis (TGA). Microscopic surface structure analyzed by Transmission Electron Microscopy (TEM) has showed spherical shapes of AgNPs with size in the range of 3-15 nm. The extent of nanosilver loading was decreased by increasing Cs content into network. The silver-loaded hydrogel was thermally more stable than the unloaded dry hydrogel counterpart. The swelling equilibrium degree (Q) and centrifuge retention capacity (CRC) in deionized water were affected by both contents of Cs and the entrapped AgNPs. The nanosilver-embedded hydrogels exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus bacteria. These comprehensive results suggest that the elaborated AgNPs-loaded nanomaterials could be used to produce valuable wound dressing.

Keywords: antibacterial activity, nanocomposites, silver nanoparticles, superabsorbent Hydrogel

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Authors: Louise Ödlund, Danica Djuric Ilic

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From a system perspective, there are several benefits of DH. A possibility to utilize the excess heat from waste incineration and biomass-based combined heat and power (CHP) production (e.g. possibility to utilize the excess heat from electricity production) are two examples. However, in a future sustainable society, the benefits of DH may be less obvious. Due to the climate changes and increased energy efficiency of buildings, the demand for space heating is expected to decrease. Due to the society´s development towards circular economy, a larger amount of the waste will be material recycled, and the possibility for DH production by the energy recovery through waste incineration will be reduced. Furthermore, the benefits of biomass-based CHP production will be less obvious since the marginal electricity production will no longer be linked to high greenhouse gas emissions due to an increased share of renewable electricity capacity in the electricity system. The purpose of the study is (1) to provide an overview of the possible development of other sectors which may influence the DH in the future and (2) to detect new business strategies which would enable for DH to adapt to the future conditions and remain competitive to alternative heat production in the future. A system approach was applied where DH is seen as a part of an integrated system which consists of other sectors as well. The possible future development of other sectors and the possible business strategies for DH producers were searched through a systematic literature review In order to remain competitive to the alternative heat production in the future, DH producers need to develop new business strategies. While the demand for space heating is expected to decrease, the space cooling demand will probably increase due to the climate changes, but also due to the better insulation of buildings in the cases where the home appliances are the heat sources. This opens up a possibility for applying DH-driven absorption cooling, which would increase the annual capacity utilization of the DH plants. The benefits of the DH related to the energy recovery from the waste incineration will exist in the future since there will always be a need to take care of materials and waste that cannot be recycled (e.g. waste containing organic toxins, bacteria, such as diapers and hospital waste). Furthermore, by operating central controlled heat pumps, CHP plants, and heat storage depending on the intermittent electricity production variation, the DH companies may enable an increased share of intermittent electricity production in the national electricity grid. DH producers can also enable development of local biofuel supply chains and reduce biofuel production costs by integrating biofuel and DH production in local DH systems.

Keywords: district heating, sustainable business strategies, sustainable development, system approach

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Authors: Hanouf A. S. Al Nuwaysir, Nadine Moubayed, Abir Ben Bacha, Islem Abid

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Consumption of waste water continues to cause significant problems for human health in both developed and developing countries. Many regulations have been implied by different world authorities controlling water quality for the presence of coliforms used as standard indicators of water quality deterioration and historically leading health protection concept. In this study, the European directive for the detection of Escherichia coli, ISO 9308-1, was applied to examine and monitor coliforms in water samples collected from Wadi Hanifa and neighboring wells, Riyadh governorate, kingdom of Saudi Arabia, which is used for irrigation and industrial purposes. Samples were taken from different locations for 8 months consecutively, chlorine concentration ranging from 0.1- 0.4 mg/l, was determined using the DPD FREE CHLORINE HACH kit. Water samples were then analyzed following the ISO protocol which relies on the membrane filtration technique (0.45µm, pore size membrane filter) and a chromogenic medium TTC, a lactose based medium used for the detection and enumeration of total coliforms and E.coli. Data showed that the number of bacterial isolates ranged from 60 to 300 colonies/100ml for well and surface water samples respectively; where higher numbers were attributed to the surface samples. Organisms which apparently ferment lactose on TTC agar plates, appearing as orange colonies, were selected and additionally cultured on EMB and MacConkey agar for a further differentiation among E.coli and coliform bacteria. Two additional biochemical tests (Cytochrome oxidase and indole from tryptophan) were also investigated to detect and differentiate the presence of E.coli from other coliforms, E. coli was identified in an average of 5 to 7colonies among 25 selected colonies.On the other hand, a more rapid, specific and sensitive analytical molecular detection namely single colony PCR was also performed targeting hha gene to sensitively detect E.coli, giving more accurate and time consuming identification of colonies considered presumptively as E.coli. Comparative methodologies, such as ultrafiltration and direct DNA extraction from membrane filters (MoBio, Grermany) were also applied; however, results were not as accurate as the membrane filtration, making it a technique of choice for the detection and enumeration of water coliforms, followed by sufficiently specific enzymatic confirmatory stage.

Keywords: coliform, cytochrome oxidase, hha primer, membrane filtration, single colony PCR

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Authors: Maximiliano Guerra, Lexi Frankel, Amalia D. Ardeljan, Sarah Ghali, Diya Kohli, Omar M. Rashid.

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Introduction/Background: Haemophilus influenzae is present as a commensal organism in the nasopharynx of most healthy adults from where it can spread to cause both systemic and respiratory tract infection. Pathogenic properties of this bacterium as well as defects in host defense may result in the spread of these bacteria throughout the body. This can result in a proinflammatory state and colonization particularly in the lungs. Recent studies have failed to determine a link between H. Influenzae colonization and prostate cancer, despite previous research demonstrating the presence of proinflammatory states in preneoplastic and neoplastic prostate lesions. Given these contradictory findings, the primary goal of this study was to evaluate the correlation between H. Influenzae infection and the incidence of prostate cancer. Methods: To evaluate the incidence of Haemophilus influenzae infection and the development of prostate cancer in the future we used data provided by a Health Insurance Portability and Accountability Act (HIPAA) compliant national database. We were afforded access to this database by Holy Cross Health, Fort Lauderdale for the express purpose of academic research. Standard statistical methods were employed in this study including Pearson’s chi-square tests. Results: Between January 2010 and December 2019, the query was analyzed and resulted in 13, 691 patients in both the control and C. difficile infected groups, respectively. The two groups were matched by age range and CCI score. In the Haemophilus influenzae infected group, the incidence of prostate cancer was 1.46%, while the incidence of the prostate cancer control group was 4.56%. The observed difference in cancer incidence was determined to be a statistically significant p-value (< 2.2x10^-16). This suggests that patients with a history of C. difficile have less risk of developing prostate cancer (OR 0.425, 95% CI: 0.382 - 0.472). Treatment bias was considered, the data was analyzed and resulted in two groups matched groups of 3,208 patients in both the infected with H. Influenzae treated group and the control who used the same medications for a different cause. Patients infected with H. Influenzae and treated had an incidence of prostate cancer of 2.49% whereas the control group incidence of prostate cancer was 4.92% with a p-value (< 2.2x10^-16) OR 0.455 CI 95% (0.526 -0.754), proving that the initial results were not due to the use of medications. Conclusion: The findings of our study reveal a statistically significant correlation between H. Influenzae infection and a decreased incidence of prostate cancer. Our findings suggest that prior infection with H. Influenzae may confer some degree of protection to patients and reduce their risk for developing prostate cancer. Future research is recommended to further characterize the potential role of Haemophilus influenzae in the pathogenesis of prostate cancer.

Keywords: Haemophilus Influenzae, incidence, prostate cancer, risk.

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Authors: L. D. Upegui, Isabel Alvarez-Solorza, Karina Garduno-Ulloa, Maren Boecker

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Introduction: The increasing prevalence rate of resistant and multiresistant bacterial strains to antibiotics is a threat to public health and requires a rapid multifunctional answer. Individuals that are affected by resistant strains present a higher morbidity and mortality than individuals that are infected with the same species of bacteria but with sensitive strains. There have been identified risk factors that are related to the misuse and overuse of antibiotics, like socio-demographic characteristics and psychological aspects of the individuals that have not been explored objectively due to a lack of valid and reliable instruments for their measurement. Objective: To validate a questionnaire for the evaluation of the levels of knowledge related to the use of antibiotics in a Mexican population. Materials and Methods: Analytical cross-sectional observational study. The questionnaire consists of 12 items to evaluated knowledge (1=no, 2=not sure, 3=yes) regarding the use of antibiotics, with higher scores corresponding to a higher level of knowledge. Data are collected in a sample of students. Data collection is still ongoing. In this abstract preliminary results of 30 respondents are reported which were collected during pilot-testing. The validation of the instrument was done using the Rasch model. Fit to the Rasch model was tested checking overall fit to the model, unidimensionality, local independence and evaluating the presence of Differential Item Functioning (DIF) by age and gender. The software Rumm2030 and the SPSS were used for the analyses. Results: The participants of the pilot-testing presented an average age of 32 years ± 12.6 and 53% were women. The preliminary results indicated that the items showed good fit to the Rasch model (chi-squared=12.8 p=0.3795). Unidimensionality (number of significant t-tests of 3%) could be proven, the items were locally independent, and no DIF was observed. Knowledge was the smallest regarding statements on the role of antibiotics in treating infections, e.g., most of the respondents did not know that antibiotics would not work against viral infections (70%) and that they could also cause side effects (87%). The knowledge score ranged from 0 to 100 points with a transformed measurement (mean of knowledge 27.1 ± 4.8). Conclusions: The instrument showed good psychometric proprieties. The low scores of knowledge about antibiotics suggest that misinterpretations on the use of these medicaments were prevalent, which could influence the production of antibiotic resistance. The application of this questionnaire will allow the objective identification of 'Hight risk groups', which will be the target population for future educational campaigns, to reduce the knowledge gaps on the general population as an effort against antibiotic resistance.

Keywords: antibiotics, knowledge, misuse, overuse, questionnaire, Rasch model, validation

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Authors: Asma Bashir, Neha Farid, Kashif Ali, Kiran Fatima

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Background: Bacteriocins are a subclass of antimicrobial peptides that are becoming extremely important in treatments. It is possible to utilise bacteriocins in place of or in addition to traditional antibiotics. It is possible to treat a variety of infections, including Vancomycin-Resistant Staphylococcus aureus (VRSA) and Methicillin-Resistant Staphylococcus aureus (MRSA), using the targeted spectrum of activity of these microorganisms. Method: This study aimed to examine the efficiency of antibiotics and bacteriocin against VRSA and MRSA. The effects of bacteriocins, such as enterocin KAE01, enterocin KAE03, enterocin KAE05, and enterocin KAE06 isolated from Enterococcus faecium strains, alone and in combination with vancomycin and methicillin antibiotics were examined. The selection technique utilized the minimum inhibitory concentrations (MICs) against Gram-positive indicator strain ATCC 6538 Methicillin-Resistant Staphylococcus aureus (MRSA) and indicator strain KSA 02 Vancomycin-Resistant Staphylococcus aureus (VRSA). Results: We report the isolation and identification of enterocins KAE01, KAE03, KAE05, and KAE06 from food isolates of Enterococcus faecium (KAE01, KAE03, KAE05, and KAE06). After isolating the protein, it was partially purified with ammonium sulphate precipitation and purified with fast protein liquid chromatography (FPLC) procedures. Combinations of enterocin KAE01, 1 citric acid, 1 lactic acid, and microcin J25, 1 reuterin, 1 citric acid, and microcin J25, 1 reuterin, 1 lactic acid shown synergistic benefits (FIC index = 0.5) against Vancomycin-Resistant Staphylococcus aureus (VRSA). In addition, a moderately synergistic (FIC index = 0.75) interaction was seen between pediocin PA-1, 1 citric acid, 1 lactic acid, and reuterin 1 citric acid, 1 lactic acid against L. ivanovii HPB28. In the presence of acids, nisin Z exhibited a modestly synergistic effect (FIC index = 0.625-0.75); however, it exhibited additive effects (FIC index = 1) when combined with reuterin or pediocin PA-1 against L. ivanovii HPB28. The efficacy of synergistic consortiums against Gram-positive bacteria was examined. Conclusion: Combining antimicrobials with various modes of action boosted efficacy and expanded the spectrum of inhibition, particularly against multidrug-resistant pathogens, according to our research.

Keywords: Enterococcus faecium, bacteriocin, antimicrobial resistance, antagonistic activity, vancomycin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus aureus

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Authors: Umut Kokbas, Mustafa Nisari

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Immunoglobulins, also known as antibodies, are glycoprotein molecules produced by activated B cells that transform into plasma cells and result in them. Antibodies are critical molecules of the immune response to fight, which help the immune system specifically recognize and destroy antigens such as bacteria, viruses, and toxins. Immunoglobulin classes differ in their biological properties, structures, targets, functions, and distributions. Five major classes of antibodies have been identified in mammals: IgA, IgD, IgE, IgG, and IgM. Evaluation of the immunoglobulin isotype can provide a useful insight into the complex humoral immune response. Evaluation and knowledge of immunoglobulin structure and classes are also important for the selection and preparation of antibodies for immunoassays and other detection applications. The immunoglobulin test measures the level of certain immunoglobulins in the blood. IgA, IgG, and IgM are usually measured together. In this way, they can provide doctors with important information, especially regarding immune deficiency diseases. Hypogammaglobulinemia (HGG) is one of the main groups of primary immunodeficiency disorders. HGG is caused by various defects in B cell lineage or function that result in low levels of immunoglobulins in the bloodstream. This affects the body's immune response, causing a wide range of clinical features, from asymptomatic diseases to severe and recurrent infections, chronic inflammation and autoimmunity Transient infant hypogammaglobulinemia (THGI), IgM deficiency (IgMD), Bruton agammaglobulinemia, IgA deficiency (SIgAD) HGG samples are a few. Most patients can continue their normal lives by taking prophylactic antibiotics. However, patients with severe infections require intravenous immune serum globulin (IVIG) therapy. The IgE level may rise to fight off parasitic infections, as well as a sign that the body is overreacting to allergens. Also, since the immune response can vary with different antigens, measuring specific antibody levels also aids in the interpretation of the immune response after immunization or vaccination. Immune deficiencies usually occur in childhood. In Immunology and Allergy clinics, apart from the classical methods, it will be more useful in terms of diagnosis and follow-up of diseases, if it is fast, reliable and especially in childhood hypogammaglobulinemia, sampling from children with a method that is more convenient and uncomplicated. The antibodies were attached to the electrode surface via the poly hydroxyethyl methacrylamide cysteine nanopolymer. It was used to evaluate the anodic peak results obtained in the electrochemical study. According to the data obtained, immunoglobulin determination can be made with a biosensor. However, in further studies, it will be useful to develop a medical diagnostic kit with biomedical engineering and to increase its sensitivity.

Keywords: biosensor, immunosensor, immunoglobulin, infection

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Authors: Nibal Al-Batsh, Issam Al-Khatib, Subha Ghannam

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Yatta is the study area for this research, located 9 km south of Hebron City in the West Bank in Palestine. It has been connected to a water network since 1974 serving nearly 85% of the households. The water network is old and inadequate to meet the needs of the population. The water supply made available to the area is also very limited, estimated to be around 20 l/c.d. Residents are thus forced to rely on water vendors which supply water with a lower quality compared to municipal water while being 400% more expensive. As a cheaper and more reliable alternative, rainwater harvesting is a common practice in the area, with the majority of the households owning at least one cistern. Rainwater harvesting is of great socio-economic importance in areas where water sources are scarce or polluted. The quality of harvested rainwater used for drinking and domestic purposes in the Yatta area was assessed throughout a year long period. A total of 100 water samples were collected from (50 rainfed cisterns) with an average capacity of 69 m3, adjacent to cement-roof catchment with an average area of 145 m2. Samples were analyzed for a number of parameters including: pH, Alkalinity, Hardness, Turbidity, Total Dissolved Solids (TDS), NO3, NH4, chloride and salinity. Microbiological contents such as Total Coliforms (TC) and Fecal Coliforms (FC) bacteria were also analyzed. Results showed that most of the rainwater samples were within WHO and EPA guidelines set for chemical parameters while revealing biological contamination. The pH values of mixed water ranged from 6.9 to 8.74 with a mean value of 7.6. collected Rainwater had lower pH values than mixed water ranging from 7.00 to 7.57 with a mean of 7.21. Rainwater also had lower average values of conductivity (389.11 µScm-1) compared to that of mixed water (463.74 µScm-1) thus indicating lower values of salinity (0.75%). The largest TDS value measured in rainwater was 316 mg/l with a mean of 199.86 mg /l. As far as microbiological quality is concerned, TC and FC were detected in 99%, 52% of collected rainwater samples, respectively. The research also addressed the impact of different socio-economic attributes on rainwater harvesting using information collected through a survey from the area. Results indicated that the majority of homeowners have the primary knowledge necessary to collect and store water in cisterns. Most of the respondents clean both the cisterns and the catchment areas. However, the research also arrives at a conclusion that cleaning is not done in a proper manner. Results show that cisterns with an operating capacity of 69 m3 would provide sufficient water to get through the dry summer months. However, the catchment area must exceed 146 m2 to produce sufficient water to fill a cistern of this size in a year receiving average precipitation.

Keywords: rainwater harvesting, runoff coefficient, water quality, microbiological contamination

Procedia PDF Downloads 285

Authors: Noah Emil Glisik

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Alcohol use disorder (AUD) presents significant health challenges worldwide and is particularly concerning in Slovenia, where high alcohol consumption contributes to elevated rates of comorbidities, including depression and suicide. This review examines emerging evidence linking gut microbiome dysbiosis to AUD, exploring whether gut microbiome alterations merely result from alcohol use or actively contribute to the persistence of addiction. Additionally, it discusses how microbial changes may influence psychological symptoms, including anxiety and depressive states, which are closely associated with suicidality in this population. To address gaps in existing research, a systematic literature search was conducted through PubMed, Web of Science, and ScienceDirect. Inclusion criteria focused on studies examining gut microbiome changes in AUD, particularly those assessing gut-brain axis interactions and microbial species impacting inflammation and neurotransmitter pathways. Studies were excluded if they lacked peer review or did not specifically assess microbiome effects on mental health outcomes. A qualitative literature review approach was applied, synthesizing findings into key themes on microbial changes, neuroinflammatory pathways, and treatment implications. Data were organized into tables to provide a clear comparison of microbiota alterations across studies, highlighting specific bacterial species and their potential effects on AUD. This review emphasizes patterns in AUD patients, where reductions in anti-inflammatory species, such as Faecalibacterium prausnitzii and Roseburia intestinalis, coincide with increases in pro-inflammatory bacteria like Enterococcus faecalisand Lactobacillus rhamnosus. These shifts contribute to increased gut permeability and systemic inflammation, potentially influencing the kynurenine pathway, which is linked to depressive symptoms and elevated alcohol cravings. Furthermore, the review explores the potential of probiotic therapies targeting these microbial imbalances as adjunctive treatments for AUD, particularly those focusing on strains that support anti-inflammatory pathways and gut barrier integrity. Restoring microbial homeostasis through probiotics or fecal microbiota transplantation may not only reduce inflammation but also alleviate mental health symptoms associated with addiction, including suicidality. The findings underscore the need for further clinical trials assessing microbiome-targeted therapies as innovative, multifaceted approaches to AUD treatment in Slovenia and beyond.

Keywords: alcohol use disorder, gut-brain axis, microbiome dysbiosis, probiotic therapy.

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Authors: Jos Olijve

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Introduction and Purpose: Endotoxins are found in the outer membrane of gram-negative bacteria and have profound in vitro and in vivo responses. They can trigger strong immune responses and negatively affect various cellar activities particular cells expressing toll-like receptors. They are therefore unwanted contaminants of biomaterials sourced from natural raw materials, and their activity must be as low as possible. Collagen and gelatin are natural extracellular matrix components and have, due to their low allergenic potential, suitable biological properties, and tunable physical characteristics, high potential in biomedical applications. The purpose of this study was to determine the influence of autoclave sterilization of gelatin on physical properties and endotoxin level compared to surfactant purified gelatin. Methods: Type A gelatin from Sigma-Aldrich (G1890) with endotoxin level of 35000 endotoxin units (EU) per gram gelatin and type A gelatins from Rousselot Gent with endotoxin activity of 30000 EU per gram were used. A 10 w/w% G1890 gelatin solution was autoclave sterilized during 30 minutes at 121°C and 1 bar over pressure. The physical properties and the endotoxin level of the sterilized G1890 gelatin were compared to a type A gelatin from Rousselot purified with Triton X100 surfactant. The Triton X100 was added to a concentration of 0.5 w/w% which is above the critical micellar concentration. The gelatin surfactant mixtures were kept for 30-45 minutes under constant stirring at 55-60°C. The Triton X100 was removed by active carbon filtration. The endotoxin levels of the gelatins were measured using the Endozyme recombinant factor C method from Hyglos GmbH (Germany). Results and Discussion: Autoclave sterilization significantly affect the physical properties of gelatin. Molecular weight of G1890 decreased from 140 to 50kDa, and gel strength decreased from 300 to 40g. The endotoxin level of the gelatin reduced after sterilization from 35000 EU/g to levels of 400-500 EU/g. These endotoxin levels are however still far above the upper endotoxin level of 0.05 EU/ml, which resembles 5 EU/g gelatin based on a 1% gelatin solution, to avoid cell proliferation alteration. Molecular weight and gel strength of Rousselot gelatin was not altered after Triton X100 purification and remained 150kDa and 300g respectively. The endotoxin levels of Triton X100 purified Rousselot gelatin was < 5EU/g gelatin. Conclusion: Autoclave sterilization of gelatin is, in comparison to Triton X100 purification, not efficient to inactivate endotoxin levels in gelatin to levels below the upper limit to avoid cell proliferation alteration. Autoclave sterilization gave a significant decrease in molecular weight and gel strength which makes autoclave sterilized gelatin, in comparison to Triton X100 purified gelatin, not suitable for 3D printing.

Keywords: endotoxin, gelatin, molecular weight, sterilization, Triton X100

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Authors: Piya Roychoudhury, Ruma Pal

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Cyanobacteria, commonly known as blue green algae were found to be an effective bioreagent for nanoparticle synthesis. Nowadays silver nanoparticles (AgNPs) are very popular due to their antimicrobial and anti-proliferative activity. To exploit these characters in different biotechnological fields, it is very essential to synthesize more stable, non-toxic nano-silver. For this reason silver-gold alloy (Ag-AuNPs) nanoparticles are of great interest as they are more stable, harder and more effective than single metal nanoparticles. In the present communication we described a simple technique for rapid synthesis of biocompatible AgNP and Ag-AuNP employing cyanobacteria, Leptolyngbya and Lyngbya respectively. For synthesis of AgNP the biomass of Leptolyngbya valderiana (200 mg Fresh weight) was exposed to 9 mM AgNO3 solution (pH 4). For synthesis of Ag-AuNP Lyngbya majuscula (200 mg Fresh weight) was exposed to equimolar solution of hydrogen tetra-auro chlorate and silver nitrate (1mM, pH 4). After 72 hrs of exposure thallus of Leptolyngyba turned brown in color and filaments of Lyngbya turned pink in color that indicated synthesis of nanoparticles. The produced particles were extracted from the cyanobacterial biomass using nano-capping agent, sodium citrate. Firstly, extracted brown and pink suspensions were taken for Energy Dispersive X-ray (EDAX) analysis to confirm the presence of silver in brown suspension and presence of both gold and silver in pink suspension. Extracted nanoparticles showed a distinct single plasmon band (AgNP at 411 nm; Ag-Au NP at 481 nm) in Uv-vis spectroscopy. It was revealed from Transmission electron microscopy (TEM) that all the synthesized particles were spherical in nature with a size range of ~2-25 nm. In X-ray powder diffraction (XRD) analysis four intense peaks appeared at 38.2°, 44.5°, 64.8°and 77.8° which confirmed the crystallographic nature of synthesized particles. Presence of different functional groups viz. N-H, C=C, C–O, C=O on the surface of nanoparticles were recorded by Fourier transform infrared spectroscopy (FTIR). Scanning Electron microscopy (SEM) images showed the surface topography of metal treated filaments of cyanobacteria. The stability of the particles was observed by Zeta potential study. Antibiotic property of synthesized particles was tested by Agar well diffusion method against gram negative bacteria Pseudomonas aeruginosa. Overall, this green-technique requires low energy, less manufacturing cost and produces rapidly eco-friendly metal nanoparticles.

Keywords: cyanobacteria, silver nanoparticles, silver-gold alloy nanoparticles, spectroscopy

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Authors: Andreea Molnar, Amalia Ardeljan, Lexi Frankel, Marissa Dallara, Brittany Nagel, Omar Rashid

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Background: Escherichia coli is a gram-negative, facultative anaerobic bacteria that belongs to the family Enterobacteriaceae and resides in the intestinal tracts of individuals. E.Coli has numerous strains grouped into serogroups and serotypes based on differences in antigens in their cell walls (somatic, or “O” antigens) and flagella (“H” antigens). More than 700 serotypes of E. coli have been identified. Although most strains of E. coli are harmless, a few strains, such as E. coli O157:H7 which produces Shiga toxin, can cause intestinal infection with symptoms of severe abdominal cramps, bloody diarrhea, and vomiting. Infection with E. Coli can lead to the development of systemic inflammation as the toxin exerts its effects. Chronic inflammation is now known to contribute to cancer development in several organs, including the prostate. The purpose of this study was to evaluate the correlation between E. Coli and the incidence of prostate cancer. Methods: Data collected in this cohort study was provided by a Health Insurance Portability and Accountability Act (HIPAA) compliant national database to evaluate patients infected with E.Coli infection and prostate cancer using the International Classification of Disease (ICD-10 and ICD-9 codes). Permission to use the database was granted by Holy Cross Health, Fort Lauderdale for the purpose of academic research. Data analysis was conducted through the use of standard statistical methods. Results: Between January 2010 and December 2019, the query was analyzed and resulted in 81, 037 patients after matching in both infected and control groups, respectively. The two groups were matched by Age Range and CCI score. The incidence of prostate cancer was 2.07% and 1,680 patients in the E. Coli group compared to 5.19% and 4,206 patients in the control group. The difference was statistically significant by a p-value p<2.2x10-16 with an Odds Ratio of 0.53 and a 95% CI. Based on the specific treatment for E.Coli, the infected group vs control group were matched again with a result of 31,696 patients in each group. 827 out of 31,696 (2.60%) patients with a prior E.coli infection and treated with antibiotics were compared to 1634 out of 31,696 (5.15%) patients with no history of E.coli infection (control) and received antibiotic treatment. Both populations subsequently developed prostate carcinoma. Results remained statistically significant (p<2.2x10-16), Odds Ratio=0.55 (95% CI 0.51-0.59). Conclusion: This retrospective study shows a statistically significant correlation between E.Coli infection and a decreased incidence of prostate cancer. Further evaluation is needed in order to identify the impact of E.Coli infection and prostate cancer development.

Keywords: E. Coli, prostate cancer, protective, microbiology

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Authors: Carolina Gutierrez-Cortes, Hector Suarez, Gustavo Buitrago

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Bacteriocins are antimicrobial peptides produced by bacteria as a competitive strategy for substrate and habitat. Those peptides have a potential use as food biopreservatives due to their antimicrobial activity against foodborne pathogens, avoiding the use of additives that can be harmful to consumers. The industrial production of bacteriocins is currently expensive; one of the options to be competitive is the development of economic culture media, for example, with the use of agro-industrial wastes such as whey. This study evaluated the growth and production of bacteriocins from four strains: Pediococcus pentosaceus 63, Pediococcus pentosaceus 145, Pediococcus pentosaceus 146 and Pediococcus pentosaceus 147 isolated from ‘minas cheese’ (artisanal cheese made from raw milk in the state of Minas Gerais, Brazil) in order to select a strain with growth at high rates and higher antimicrobial activity against Listeria monocytogenes 104 after incubation on the culture medium designed with whey and other components. The media used were: MRS broth, modified MRS broth (using different sources of carbon and nitrogen and different amounts of micronutrients) and a culture medium designed by a factorial design using whey and other components. The final biomass concentrations of the four strains in MRS broth after 24 hours of incubation were very similar 9.25, 9.33, 9.25 and 9.22 (log CFU/mL) for P. pentosaceus 63, P. pentosaceus 145, P. pentosaceus 146 and P. pentosaceus 147 respectively. In the same assays, antimicrobial activity of 3200 AU/mL for the first three and of 12800 AU/mL for P. pentosaceus 147 were obtained. Culture of P. pentosaceus 63 on modified MRS broth, showed the effect of some sources of carbon on the activity of bacteriocin, obtaining 12800 AU/mL with dextrose and 25600 AU/mL with maltose. Cultures of P. pentosaceus 145, 146 and 147 with these same sugars presented activity of 12800 AU/mL. It was observed that the modified MRS medium using whey increased the antimicrobial activity of the strains at 16000, 6400, 16000 and 19200 AU/mL for each strain respectively, keeping the biomass at values close to 9 log units. About nitrogen sources, it was observed that the combination of peptone (10 g /L), meat extract (10 g/L) and yeast extract (5 g/L) promoted the highest activity (12800 AU/mL), and in all cases MgSO4, MnSO4, K2HPO4 and ammonium citrate at low concentrations adversely affected bacteriocin production. Because P. pentosaceus 147 showed the highest antimicrobial activity in the presence of whey, it was used to evaluate the culture medium (peptone (10 g/L), meat extract (8 g/L), yeast extract (2 g/L), Tween® 80 (1 g/L), ammonium citrate (2 g/L), sodium acetate (5 g/L), MgSO4 (0.2 g/L), MnSO4 (0.04 g/L)). With the designed medium added with whey, 9.34 log units of biomass concentration and 19200 AU/mL were achieved for P. pentosaceus 147. The above suggest that the new medium promotes the antimicrobial activity of P. pentosaceus 147 allowing the use of an economic medium using whey.

Keywords: antimicrobial activity, bacteriocins, pediococcus, whey

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Authors: Rym Saidi, Pape Alioune Ndiaye, Mohamed Idbella, Ammar Ibnyasser, Zineb Rchiad, Issam Kadmiri Meftahi, Khalid Daoui, Adnane Bargaz

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Chickpea (Cicer arietinum L.) is an important leguminous crop grown worldwide, and the second most important food legume in Morocco. In addition, that chickpea plays a significant role in humans’ dietary consumption, it has key ecological interest in terms of biological N-fixation (BNF) having the ability to symbiotically secure 20-80% of needed. Alongside nitrogen (N), low soil phosphorus (P) availability is one of the major factors limiting chickpea growth and productivity. After nitrogen, P is the most important macronutrient for plants growth and development as well as the BNF. In the context of improving chickpea symbiotic performance, co-application of beneficial bacterial inoculants (including Mesorhizobium) and Rock P-fertilizer could boost chickpea performance and productivity, owing to increasing P-utilization efficiency and overall nutrient acquisition under P-deficiency conditions. Greenhouse experiment was conducted to evaluate the response of two chickpea varieties (Arifi “A” and Bochra “B”) to co-application of RP-fertilizer alongside Mesorhizobium and phosphate solubilizing bacteria (PSB) consortium under P-deficient soil in Morocco. Our findings demonstrate that co-applying RP50 with bacterial inoculant significantly increased NDW by 85.71% and 109.09% in A and B chickpea varieties respectively, compared to uninoculated RP-fertilized plants. Nodule Pi and leghemoglobin (LHb) contents also increased in RP-fertilized bacterial inoculants plants. Likewise, shoot and root dry weights of both chickpea varieties increased with bacterial inoculation and RP-fertilization. This is due to enhanced Pi content in shoot (282.54% and 291.42%) and root (334.30% and 408.32%) in response to RP50-Inc compared to unfertilized uninoculated plants, for A and B chickpea varieties respectively. Rhizosphere available P was also increased by 173.86% and 182.25% in response to RP50-Inc as compared to RP-fertilized uninoculated plants, with a positive correlation between soil available P and root length in inoculated plants of A. and B. chickpea varieties (R= 0.49; 0.6) respectively. Furthermore, Mesorhizobium was among the dominant genera in rhizosphere bacterial diversity of both chickpea varieties. This can be attributed to its capacity to enhance plant growth traits, with a more pronounced effect observed in B. variety. Our research demonstrates that integrated fertilization with bacterial inoculation effectively improves biological N-fixation and P nutrition, enhancing the agrophysiological performance of Moroccan chickpea varieties, particularly in restricted P-availability conditions.

Keywords: chickpea varieties, bacterial consortium, inoculants, Mesorhizobium, Rock-P fertilizer, phosphorus deficiency, agrophysiological performance

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Authors: Wael M. Rabeh, Cesar Carrasco-Lopez, Juliana C. Ferreira, Pance Naumov

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Bioluminescence, the emission of light from a biological process, is found in various living organisms including bacteria, fireflies, beetles, fungus and different marine organisms. Luciferase is an enzyme that catalyzes a two steps oxidation of luciferin in the presence of Mg2+ and ATP to produce oxyluciferin and releases energy in the form of light. The luciferase assay is used in biological research and clinical applications for in vivo imaging, cell proliferation, and protein folding and secretion analysis. The luciferase enzyme consists of two domains, a large N-terminal domain (1-436 residues) that is connected to a small C-terminal domain (440-544) by a flexible loop that functions as a hinge for opening and closing the active site. The two domains are separated by a large cleft housing the active site that closes after binding the substrates, luciferin and ATP. Even though all insect luciferases catalyze the same chemical reaction and share 50% to 90% sequence homology and high structural similarity, they emit light of different colors from green at 560nm to red at 640 nm. Currently, the majority of the structural and biochemical studies have been conducted on green-emitting firefly luciferases. To address the color emission mechanism, we expressed and purified two luciferase enzymes with blue-shifted green and red emission from indigenous Brazilian species Amydetes fanestratus and Phrixothrix, respectively. The two enzymes naturally emit light of different colors and they are an excellent system to study the color-emission mechanism of luciferases, as the current proposed mechanisms are based on mutagenesis studies. Using a vapor-diffusion method and a high-throughput approach, we crystallized and solved the crystal structure of both enzymes, at 1.7 Å and 3.1 Å resolution respectively, using X-ray crystallography. The free enzyme adopted two open conformations in the crystallographic unit cell that are different from the previously characterized firefly luciferase. The blue-shifted green luciferase crystalized as a monomer similar to other luciferases reported in literature, while the red luciferases crystalized as an octamer and was also purified as an octomer in solution. The octomer conformation is the first of its kind for any insect’s luciferase, which might be relate to the red color emission. Structurally designed mutations confirmed the importance of the transition between the open and close conformations in the fine-tuning of the color and the characterization of other interesting mutants is underway.

Keywords: bioluminescence, enzymology, structural biology, x-ray crystallography

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Authors: Isabel Brás, Artur Figueirinha, Bruno Esteves, Luísa P. Cruz-Lopes

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The agriculture lignocellulosic by-products are receiving increased attention, namely in the search for filter materials that retain contaminants from water. These by-products, specifically almond and hazelnut shells are abundant in Portugal once almond and hazelnuts production is a local important activity. Hazelnut and almond shells have as main constituents lignin, cellulose and hemicelluloses, water soluble extractives and tannins. Along the adsorption of heavy metals from contaminated waters, water soluble compounds can leach from shells and have a negative impact in the environment. Usually, the chemical characterization of treated water by itself may not show environmental impact caused by the discharges when parameters obey to legal quality standards for water. Only biological systems can detect the toxic effects of the water constituents. Therefore, the evaluation of toxicity by biological tests is very important when deciding the suitability for safe water discharge or for irrigation applications. The main purpose of the present work was to assess the potential impacts of waters after been treated for heavy metal removal by hazelnut and almond shells adsorption systems, with short term acute toxicity tests. To conduct the study, water at pH 6 with 25 mg.L-1 of lead, was treated with 10 g of shell per litre of wastewater, for 24 hours. This procedure was followed for each bark. Afterwards the water was collected for toxicological assays; namely bacterial resistance, seed germination, Lemna minor L. test and plant grow. The effect in isolated bacteria strains was determined by disc diffusion method and the germination index of seed was evaluated using lettuce, with temperature and humidity germination control for 7 days. For aquatic higher organism, Lemnas were used with 4 days contact time with shell solutions, in controlled light and temperature. For terrestrial higher plants, biomass production was evaluated after 14 days of tomato germination had occurred in soil, with controlled humidity, light and temperature. Toxicity tests of water treated with shells revealed in some extent effects in the tested organisms, with the test assays showing a close behaviour as the control, leading to the conclusion that its further utilization may not be considered to create a serious risk to the environment.

Keywords: lignocellulosic wastes, adsorption, acute toxicity tests, risk assessment

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Authors: Eliane G. Tótoli, Hérida Regina N. Salgado

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Daptomycin is an important antimicrobial agent used in clinical practice nowadays, since it is very active against some Gram-positive bacteria that are particularly challenges for the medicine, such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE). The importance of environmental preservation has receiving special attention since last years. Considering the evident need to protect the natural environment and the introduction of strict quality requirements regarding analytical procedures used in pharmaceutical analysis, the industries must seek environmentally friendly alternatives in relation to the analytical methods and other processes that they follow in their routine. In view of these factors, green analytical chemistry is prevalent and encouraged nowadays. In this context, infrared spectroscopy stands out. This is a method that does not use organic solvents and, although it is formally accepted for the identification of individual compounds, also allows the quantitation of substances. Considering that there are few green analytical methods described in literature for the analysis of daptomycin, the aim of this work was the development and validation of a green analytical method for the quantification of this drug in lyophilized powder for injectable solution, by Fourier-transform infrared spectroscopy (FT-IR). Method: Translucent potassium bromide pellets containing predetermined amounts of the drug were prepared and subjected to spectrophotometric analysis in the mid-infrared region. After obtaining the infrared spectrum and with the assistance of the IR Solution software, quantitative analysis was carried out in the spectral region between 1575 and 1700 cm-1, related to a carbonyl band of the daptomycin molecule, and this band had its height analyzed in terms of absorbance. The method was validated according to ICH guidelines regarding linearity, precision (repeatability and intermediate precision), accuracy and robustness. Results and discussion: The method showed to be linear (r = 0.9999), precise (RSD% < 2.0), accurate and robust, over a concentration range from 0.2 to 0.6 mg/pellet. In addition, this technique does not use organic solvents, which is one great advantage over the most common analytical methods. This fact contributes to minimize the generation of organic solvent waste by the industry and thereby reduces the impact of its activities on the environment. Conclusion: The validated method proved to be adequate to quantify daptomycin in lyophilized powder for injectable solution and can be used for its routine analysis in quality control. In addition, the proposed method is environmentally friendly, which is in line with the global trend.

Keywords: daptomycin, Fourier-transform infrared spectroscopy, green analytical chemistry, quality control, spectrometry in IR region

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Authors: Jeffrey A. Amelse

Abstract:

Many corporations aspire to become Net Zero Carbon Carbon Dioxide by 2035-2050. This paper examines what it will take to achieve those goals. Achieving Net Zero CO₂ requires an understanding of where energy is produced and consumed, the magnitude of CO₂ generation, and proper understanding of the Carbon Cycle. The latter leads to the distinction between CO₂ and biomass carbon sequestration. Short reviews are provided for prior technologies proposed for reducing CO₂ emissions from fossil fuels or substitution by renewable energy, to focus on their limitations and to show that none offer a complete solution. Of these, CO₂ sequestration is poised to have the largest impact. It will just cost money, scale-up is a huge challenge, and it will not be a complete solution. CO₂ sequestration is still in the demonstration and semi-commercial scale. Transportation accounts for only about 30% of total U.S. energy demand, and renewables account for only a small fraction of that sector. Yet, bioethanol production consumes 40% of U.S. corn crop, and biodiesel consumes 30% of U.S. soybeans. It is unrealistic to believe that biofuels can completely displace fossil fuels in the transportation market. Bioethanol is traced through its Carbon Cycle and shown to be both energy inefficient and inefficient use of biomass carbon. Both biofuels and CO₂ sequestration reduce future CO₂ emissions from continued use of fossil fuels. They will not remove CO₂ already in the atmosphere. Planting more trees has been proposed as a way to reduce atmospheric CO₂. Trees are a temporary solution. When they complete their Carbon Cycle, they die and release their carbon as CO₂ to the atmosphere. Thus, planting more trees is just 'kicking the can down the road.' The only way to permanently remove CO₂ already in the atmosphere is to break the Carbon Cycle by growing biomass from atmospheric CO₂ and sequestering biomass carbon. Sequestering tree leaves is proposed as a solution. Unlike wood, leaves have a short Carbon Cycle time constant. They renew and decompose every year. Allometric equations from the USDA indicate that theoretically, sequestrating only a fraction of the world’s tree leaves can get the world to Net Zero CO₂ without disturbing the underlying forests. How can tree leaves be permanently sequestered? It may be as simple as rethinking how landfills are designed to discourage instead of encouraging decomposition. In traditional landfills, municipal waste undergoes rapid initial aerobic decomposition to CO₂, followed by slow anaerobic decomposition to methane and CO₂. The latter can take hundreds to thousands of years. The first step in anaerobic decomposition is hydrolysis of cellulose to release sugars, which those who have worked on cellulosic ethanol know is challenging for a number of reasons. The key to permanent leaf sequestration may be keeping the landfills dry and exploiting known inhibitors for anaerobic bacteria.

Keywords: carbon dioxide, net zero, sequestration, biomass, leaves

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Authors: masmoudi Jabri Khaoula, Zitouni Hana, Bousselmi Latifa, Akrout Hanen

Abstract:

Mathematical modeling has become an essential tool for sustainable wastewater management, particularly for the simulation and the optimization of complex processes involved in activated sludge systems. In this context, the activated sludge model (ASM3h) was used for the simulation of a Biological Membrane Reactor (MBR) as it includes the integration of biological wastewater treatment and physical separation by membrane filtration. In this study, the MBR with a useful volume of 12.5 L was fed continuously with poultry slaughterhouse wastewater (PSWW) for 50 days at a feed rate of 2 L/h and for a hydraulic retention time (HRT) of 6.25h. Throughout its operation, High removal efficiency was observed for the removal of organic pollutants in terms of COD with 84% of efficiency. Moreover, the MBR has generated a treated effluent which fits with the limits of discharge into the public sewer according to the Tunisian standards which were set in March 2018. In fact, for the nitrogenous compounds, average concentrations of nitrate and nitrite in the permeat reached 0.26±0.3 mg. L-1 and 2.2±2.53 mg. L-1, respectively. The simulation of the MBR process was performed using SIMBA software v 5.0. The state variables employed in the steady state calibration of the ASM3h were determined using physical and respirometric methods. The model calibration was performed using experimental data obtained during the first 20 days of the MBR operation. Afterwards, kinetic parameters of the model were adjusted and the simulated values of COD, N-NH4+and N- NOx were compared with those reported from the experiment. A good prediction was observed for the COD, N-NH4+and N- NOx concentrations with 467 g COD/m³, 110.2 g N/m³, 3.2 g N/m³ compared to the experimental data which were 436.4 g COD/m³, 114.7 g N/m³ and 3 g N/m³, respectively. For the validation of the model under dynamic simulation, the results of the experiments obtained during the second treatment phase of 30 days were used. It was demonstrated that the model simulated the conditions accurately by yielding a similar pattern on the variation of the COD concentration. On the other hand, an underestimation of the N-NH4+ concentration was observed during the simulation compared to the experimental results and the measured N-NO3 concentrations were lower than the predicted ones, this difference could be explained by the fact that the ASM models were mainly designed for the simulation of biological processes in the activated sludge systems. In addition, more treatment time could be required by the autotrophic bacteria to achieve a complete and stable nitrification. Overall, this study demonstrated the effectiveness of mathematical modeling in the prediction of the performance of the MBR systems with respect to organic pollution, the model can be further improved for the simulation of nutrients removal for a longer treatment period.

Keywords: activated sludge model (ASM3h), membrane bioreactor (MBR), poultry slaughter wastewater (PSWW), reuse

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Authors: Gabriela Kalcikova, Igor Bosevski, Ula Rozman, Andreja Zgajnar Gotvajn

Abstract:

Antibiotics are extensively used in human medicine and also in animal husbandry to prevent or control infections. Recently, a lot of attention has been put on veterinary antibiotics, because their global consumption is increasing and it is expected to be 106.600 tons in 2030. Most of veterinary antibiotics are introduced into the environment via animal manure, which is used as fertilizer. One of such veterinary antibiotics is tiamulin. It is used the form of fumarate for treatment of pig and poultry. It is used against prophylaxis of dysentery, pneumonia and mycroplasmal infections, but its environmental impact is practically unknown. Tiamulin has been found very persistent in animal manure and thus it is expected that can be, during rainfalls, transported into the aquatic environment and affect various organisms. For assessment of its environmental impact, it is necessary to evaluate its biodegradability and toxicity to various organisms from different levels of a food chain. Therefore, the aim of our study was to evaluate ready biodegradability and toxicity of tiamulin fumarate to various organisms. Bioassay used included luminescent bacterium Vibrio fischeri heterotrophic and nitrifying microorganisms of activated sludge, water flea Daphnia magna and duckweed Lemna minor. For each species, EC₅₀ values were calculated. Biodegradability test was used for determination of ready biodegradability and it provides information about biodegradability of tiamulin under the most common environmental conditions. Results of our study showed that tiamulin differently affects selected organisms. The most sensitive organisms were water fleas with 48hEC₅₀ = 14.2 ± 4.8 mg/L and duckweed with 168hEC₅₀ = 22.6 ± 0.8 mg/L. Higher concentrations of tiamulin (from 10 mg/L) significantly affected photosynthetic pigments content in duckweed and concentrations above 80 mg/L cause visible chlorosis. It is in agreement with previous studies showing significant effect of tiamulin on green algae and cyanobacteria. Tiamuline has a low effect on microorganisms. The lower toxicity was observed for heterotrophic microorganisms (30minEC₅₀ = 1656 ± 296 mg/L), than Vibrio fisheri (30minEC₅₀ = 492 ± 21) and the most sensitive organisms were nitrifying microorganisms (30minEC₅₀ = 183 ± 127 mg/L). The reason is most probably the mode of action of tiamulin being effective to gram-positive bacteria while gram-negative (e.g., Vibrio fisheri) are more tolerant to tiamulin. Biodegradation of tiamulin was very slow with a long lag-phase being 20 days. The maximal degradation reached 40 ± 2 % in 43 days of the test and tiamulin as other antibiotics (e.g. ciprofloxacin) are not easily biodegradable. Tiamulin is widely used antibiotic in veterinary medicine and thus present in the environment. According to our results, tiamulin can have negative effect on water fleas and duckweeds, but the concentrations are several magnitudes higher than that found in any environmental compartment. Tiamulin is low toxic to tested microorganisms, but it is very low biodegradable and thus possibly persistent in the environment.

Keywords: antibiotics, biodegradability, tiamulin, toxicity

Procedia PDF Downloads 186

Authors: Yik Sin Chan, Bee Ling Chuah, Wei Quan Chan, Ri Jin Cheng, Yan Hang Oon, Kong Soo Khoo, Nam Weng Sit

Abstract:

Traditionally, medicinal plants have been used to treat different kinds of ailments including infectious diseases. They serve as a good source of lead compounds for the development of new and safer anti-infective agents. This study aimed to investigate the antimicrobial potential of the leaves of three medicinal plants, namely Catunaregam spinosa (Rubiaceae; Mountain pomegranate), Houttuynia cordata (Saururaceae; "fishy-smell herb") and Rhapis excelsa (Arecaceae; “broadleaf lady palm”). The leaves extracts were obtained by sequential extraction using hexane, chloroform, ethyl acetate, ethanol, methanol and water. The antibacterial and antifungal activities were assessed using a colorimetric broth microdilution method against a panel of human pathogenic bacteria (Gram-positive: Bacillus cereus and Staphylococcus aureus; Gram-negative: Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa) and fungi (yeasts: Candida albicans, Candida parapsilosis and Cryptococcus neoformans; Moulds: Aspergillus fumigatus and Trichophyton mentagrophytes) respectively; while antiviral activity was evaluated against the Chikungunya virus on monkey kidney epithelial (Vero) cells by neutral red uptake assay. All the plant extracts showed bacteriostatic activity, however, only 72% of the extracts (13/18) were found to have bactericidal activity. The lowest minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were given by the hexane extract of C. spinosa against S. aureus with the values of 0.16 and 0.31 mg/mL respectively. All the extracts also possessed fungistatic activity. Only the hexane, chloroform and ethyl acetate extracts of H. cordata exerted inhibitory activity against A. fumigatus, giving the lowest fungal susceptibility index of 16.7%. In contrast, only 61% of the extracts (11/18) showed fungicidal activity. The ethanol extract of R. excelsa exhibited the strongest fungicidal activity against C. albicans, C. parapsilosis and T. mentagrophytes with minimum fungicidal concentration (MFC) values of 0.04–0.08 mg/mL, in addition to its methanol extract against T. mentagrophytes (MFC=0.02 mg/mL). For anti-Chikungunya virus activity, only chloroform and ethyl acetate extracts of R. excelsa showed significant antiviral activity with 50% effective concentrations (EC50) of 29.9 and 78.1 g/mL respectively. Extracts of R. excelsa warrant further investigations into their active principles responsible for antifungal and antiviral properties.

Keywords: bactericidal, Chikungunya virus, extraction, fungicidal

Procedia PDF Downloads 403