Search results for: cellulose-degrading bacteria

Authors: Cynthia K. Burzell

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Aequor's Founder, a Marine and Medical Microbiologist, discovered novel, non-toxic chemicals in the ocean that uniquely remove biofilm in minutes and prevent its formation for days. These chemicals and over 70 synthesized analogs that Aequor developed can replace thousands of toxic biocides used in consumer and industrial products and, as new drug candidates, kill biofilm-forming bacteria and fungi Superbugs -the antimicrobial-resistant (AMR) pathogens for which there is no cure. Cynthia Burzell, PhD., is a Marine and Medical Microbiologist studying natural mechanisms that inhibit biofilm formation on surfaces in contact with water. In 2002, she discovered a new genus and several new species of marine microbes that produce small molecules that remove biofilm in minutes and prevent its formation for days. The molecules include new antimicrobials that can replace thousands of toxic biocides used in consumer and industrial products and can be developed into new drug candidates to kill the biofilm-forming bacteria and fungi -- including the antimicrobial-resistant (AMR) Superbugs for which there is no cure. Today, Aequor has over 70 chemicals that are divided into categories: (1) Novel natural chemicals. Lonza validated that the primary natural chemical removed biofilm in minutes and stated: "Nothing else known can do this at non-toxic doses." (2) Specialty chemicals. 25 of these structural analogs are already approved under the U.S. Environmental Protection Agency (EPA)'s Toxic Substances Control Act, certified as "green" and available for immediate sale. These have been validated for the following agro-industrial verticals: (a) Surface cleaners: The U.S. Department of Agriculture validated that low concentrations of Aequor's formulations provide deep cleaning of inert, nano and organic surfaces and materials; (b) Water treatments: NASA validated that one dose of Aequor's treatment in the International Space Station's water reuse/recycling system lasted 15 months without replenishment. DOE validated that our treatments lower energy consumption by over 10% in buildings and industrial processes. Future validations include pilot projects with the EPA to test efficacy in hospital plumbing systems. (c) Algae cultivation and yeast fermentation: The U.S. Department of Energy (DOE) validated that Aequor's treatment boosted biomass of renewable feedstocks by 40% in half the time -- increasing the profitability of biofuels and biobased co-products. DOE also validated increased yields and crop protection of algae under cultivation in open ponds. A private oil and gas company validated decontamination of oilfield water. (3) New structural analogs. These kill Gram-negative and Gram-positive bacteria and fungi alone, in combinations with each other, and in combination with low doses of existing, ineffective antibiotics (including Penicillin), "potentiating" them to kill AMR pathogens at doses too low to trigger resistance. Both the U.S. National Institutes for Health (NIH) and Department of Defense (DOD) has executed contracts with Aequor to provide the pre-clinical trials needed for these new drug candidates to enter the regulatory approval pipelines. Aequor seeks partners/licensees to commercialize its specialty chemicals and support to evaluate the optimal methods to scale-up of several new structural analogs via activity-guided fractionation and/or biosynthesis in order to initiate the NIH and DOD pre-clinical trials.

Keywords: biofilm, potentiation, prevention, removal

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Authors: Lucas B. Naves, Maria José Abreu

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Reprocessing single-use device has attracted interesting on the medical environment over the last decades. The reprocessing technique was sought in order to reduce the cost of purchasing the new medical device, which can achieve almost double of the price of the reprocessed product. In this manuscript, we have done a literature review, aiming the reuse of medical device that was firstly designed for single use only, but has become, more and more, effective on its reprocessing procedure. We also show the regulation, the countries which allows this procedure, the classification of these device and also the most important issue concerning the re-utilization of medical device, how to minimizing the risk of gram positive and negative bacteria, avoid cross-contamination, hepatitis B (HBV), and C (HCV) virus, and also human immunodeficiency virus (HIV).

Keywords: reusing, reprocessing, single-use medical device, HIV, hepatitis B and C

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Authors: Karima Oldyerou, B. Meddah, A. Tirtouil

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Laurusnobilis is an aromatic plant, common in Algeria and widely used by local people as a source of spice and for medicinal purposes. The essential oil of this plant is the subject of this work in a physicochemical and microbiological study. The extraction of the essential oil was carried by steam distillation and the highest yield (1.5%) was determined in May. The organoleptic and physico-chemical characters are consistent with those obtained in the literature with some differences that can be attributed to certain factors. Evaluation of antibacterial activity showed a sensitivity of Salmonella spp. with an MIC of 2,5 mg.ml-1, and other bacteria of the intestinal flora of Wistar rats: E. coli and Lactobacillus sp. have a high potential for resistance with MICs respectively equal to 10 and 20 mg.ml-1.

Keywords: laurus nobilis, essential oil, physicochemical character, MIC, intestinal flora, antibacterial activity

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Authors: D. Hammad, D. P. Tonge

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Rheumatoid arthritis (RA) is a disabling and common autoimmune disease during which the body's immune system attacks healthy tissues. This results in complicated and long-lasting actions being carried out by the immune system, which typically only occurs when the immune system encounters a foreign object. In the case of RA, the disease affects millions of people and causes joint inflammation, ultimately leading to the destruction of cartilage and bone. Interestingly, the disease mechanism still remains unclear. It is likely that RA occurs as a result of a complex interplay of genetic and environmental factors including an imbalance in the microorganism population inside our body. The human microbiome or microbiota is an extensive community of microorganisms in and on the bodies of animals, which comprises bacteria, fungi, viruses, and protozoa. Recently, the development of molecular techniques to characterize entire bacterial communities has renewed interest in the involvement of the microbiome in the development and progression of RA. We believe that an imbalance in some of the specific bacterial species in the gut, mouth and other sites may lead to atopobiosis; the translocation of these organisms into the blood, and that this may lead to changes in immune system status. The aim of this study was, therefore, to characterize the microbiome of RA serum samples in comparison to healthy control subjects using 16S rRNA gene amplification and sequencing. Serum samples were obtained from healthy control volunteers and from patients with RA both prior to, and following treatment. The bacterial community present in each sample was identified utilizing V4 region 16S rRNA amplification and sequencing. Bacterial identification, to the lowest taxonomic rank, was performed using a range of bioinformatics tools. Significantly, the proportions of the Lachnospiraceae, Ruminococcaceae, and Halmonadaceae families were significantly increased in the serum of RA patients compared with healthy control serum. Furthermore, the abundance of Bacteroides and Lachnospiraceae nk4a136_group, Lachnospiraceae_UGC-001, RuminococcaceaeUCG-014, Rumnococcus-1, and Shewanella was also raised in the serum of RA patients relative to healthy control serum. These data support the notion of a blood microbiome and reveal RA-associated changes that may have significant implications for biomarker development and may present much-needed opportunities for novel therapeutic development.

Keywords: blood microbiome, gut and oral bacteria, Rheumatoid arthritis, 16S rRNA gene sequencing

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Authors: Mohammad Reza Naghavi, Farzad Alaeimoghadam, Hossein Ghafoori

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Artemisia species, which are medically beneficial, are widespread in temperate regions of both Northern and Southern hemispheres among which Iran is located. About 35 species of Artemisia are indigenous in Iran among them some are widespread in all or most provinces, yet some are restricted to some specific regions. In this review paper, initially, GC-Mass results of some experiments done in different provinces of Iran are mentioned among them some compounds are common among species, some others are mostly restricted to other species; after that, medical advantages based on some researches on species of this genus are reviewed; different qualities such as anti-leishmania, anti-bacteria, antiviral as well as anti-proliferative could be mentioned.

Keywords: artemisia, GC-Mass analysis, medical advantage, antiviral

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Authors: Omaima A. Sharaf, Tarek A. Moussa, Said M. Badr El-Din, H. Moawad

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Background: Polycyclic aromatic hydrocarbons (PAHs) pose great environmental and human health concerns for their widespread occurrence, persistence, and carcinogenic properties. PAHs releases due to anthropogenic activities to the wider environment have led to higher concentrations of these contaminants than would be expected from natural processes alone. This may result in a wide range of environmental problems that can accumulate in agricultural ecosystems, which threatened to become a negative impact on sustainable agricultural development. Thus, this study aimed to evaluate the physico-chemical, and microbial properties of the compost leachate (CL) to assess its role as nutrient and microbial source (biostimulation/bioaugmentation) for developing a cost-effective bioremediation technology for PAHs contaminated sites. Material and Methods: PAHs-degrading bacteria were isolated from CL that was collected from a composting site located in central Scotland, UK. Isolation was carried out by enrichment using phenanthrene (PHR), pyrene (PYR) and benzo(a)pyrene (BaP) as the sole source of carbon and energy. The isolates were characterized using a variety of phenotypic and molecular properties. Six different isolates were identified based on the difference in morphological and biochemical tests. The efficiency of these isolates in PAHs utilization was assessed. Further analysis was performed to define taxonomical status and phylogenic relation between the most potent PAHs-utilizing bacterial strains and other standard strains, using molecular approach by partial 16S rDNA gene sequence analysis. Results indicated that the 16S rDNA sequence analysis confirmed the results of biochemical identification, as both of biochemical and molecular identification of the isolates assigned them to Bacillus licheniformis, Pseudomonas aeruginosa, Alcaligenes faecalis, Serratia marcescens, Enterobacter cloacae and Providenicia which were identified as the prominent PAHs-utilizers isolated from CL. Conclusion: This study indicates that the CL samples contain a diverse population of PAHs-degrading bacteria and the use of CL may have a potential for bioremediation of PAHs contaminated sites.

Keywords: polycyclic aromatic hydrocarbons, physico-chemical analyses, compost leachate, microbial and biochemical analyses, phylogenic relations, 16S rDNA sequence analysis

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Authors: Miaomiao Zhang, Sabrina Beckmann, Haluk Ertan, Rocky Chau, Mike Manefield

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Phenazines are a large class of nitrogen-containing aromatic heterocyclic compounds, which are almost exclusively produced by bacteria from diverse genera including Pseudomonas and Streptomyces. Phenazine-1-carboxylic acid (PCA) as one of 'core' phenazines are converted from chorismic acid before modified to other phenazine derivatives in different cells. Phenazines have attracted enormous interests because of their multiple roles on biocontrol, bacterial interaction, biofilm formation and fitness of their producers. However, in spite of ecological importance, degradation as a part of phenazines’ fate only have extremely limited attention now. Here, to isolate PCA-degrading bacteria, 200 mg L-1 PCA was supplied as sole carbon, nitrogen and energy source in minimal mineral medium. Quantitative PCR and Reverse-transcript PCR were employed to study abundance and activity of functional gene MFORT 16269 in PCA degradation, respectively. Intermediates and products of PCA degradation were identified with LC-MS/MS. After enrichment and isolation, a PCA-degrading strain was selected from soil and was designated as Rhodanobacter sp. PCA2 based on full 16S rRNA sequencing. As determined by HPLC, strain PCA2 consumed 200 mg L-1 (836 µM) PCA at a rate of 17.4 µM h-1, accompanying with significant cells yield from 1.92 × 105 to 3.11 × 106 cells per mL. Strain PCA2 was capable of degrading other phenazines as well, including phenazine (4.27 µM h-1), pyocyanin (2.72 µM h-1), neutral red (1.30 µM h-1) and 1-hydroxyphenazine (0.55 µM h-1). Moreover, during the incubation, transcript copies of MFORT 16269 gene increased significantly from 2.13 × 106 to 8.82 × 107 copies mL-1, which was 2.77 times faster than that of the corresponding gene copy number (2.20 × 106 to 3.32 × 107 copies mL-1), indicating that MFORT 16269 gene was activated and played roles on PCA degradation. As analyzed by LC-MS/MS, decarboxylation from the ring structure was determined as the first step of PCA degradation, followed by cleavage of nitrogen-containing ring by dioxygenase which catalyzed phenazine to nitrosobenzene. Subsequently, phenylhydroxylamine was detected after incubation for two days and was then transferred to aniline and catechol. Additionally, genomic and proteomic analyses were also carried out for strain PCA2. Overall, the findings presented here showed that a newly isolated strain Rhodanobacter sp. PCA2 was capable of degrading phenazines through decarboxylation and cleavage of nitrogen-containing ring, during which MFORT 16269 gene was activated and played important roles.

Keywords: decarboxylation, MFORT16269 gene, phenazine-1-carboxylic acid degradation, Rhodanobacter sp. PCA2

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Authors: Elmo Thiago Lins Cöuras Ford, Valentina Alessandra Carvalho do Vale

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It presents a heating system using low cost alternative solar collectors to promote the disinfection of water in low income communities that take water contaminated by bacteria. The system consists of two solar collectors, with total area of 4 m² and was built using PET bottles and cans of beer and soft drinks. Each collector is made up of 8 PVC tubes, connected in series and work in continuous flow. It will determine the flux the most appropriate to generate the temperature to promote the disinfection. Will be presented results of the efficiency and thermal loss of system and results of analysis of water after undergoing the process of heating.

Keywords: disinfection of water, solar heating system, poor communities, PVC

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Authors: Zohreh Bayat, Dariush Minai-Tehrani

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Lipases constitute one of the most important groups of industrial enzymes that catalyze the hydrolysis of triacylglycerol to glycerol and fatty acids. Muscarinic antagonist relieves smooth muscle spasm of the gastrointestinal tract and effect on the cardiovascular system. In this research, the effect of a muscarinic antagonist on the lipase activity of Pseudomonas aeruginosa was studied. Lineweaver–Burk plot showed that the drug inhibited the enzyme by competitive inhibition. The IC50 value (60 uM) and Ki (30 uM) of the drug revealed the drug bound to the enzyme with high affinity. Determination of enzyme activity in various pH and temperature showed that the maximum activity of lipase was at pH 8 and 60°C both in presence and absence of the drug.

Keywords: bacteria, inhibition, kinetics, lipase

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Authors: Khosro Ghazvinian, Touba Khodaiean

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Anaplasma organisms are obligatory intracellular bacteria belonging to the order Rickettsiales, family Anaplasmataceae. This disease is distributed around the globe and infected ticks are the most important vectors in anaplasmosis transmission. There is a little information about anaplasmosis in camels. This research investigated the blood films of 35 (20 male, 15 female) camels randomly selected from a flock of 150 camels. Samples were stained with Giemsa and Anaplasma sp. organisms were observed in six out of 35 (17.14 %) blood films. There were also some changes in Diff-Quick and morphology of leukocytes. No significant difference between male and female camels was observed (P>0.05). According to the results anaplasmosis is presented among camels in Iran.

Keywords: anaplasma, anaplasmosis, camel, Iran

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Authors: Vanja Vlajkov, Ivana PajčIn, Mila Grahovac, Marta Loc, Dragana Budakov, Jovana Grahovac

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The rhizosphere soil refers to the plant roots' dynamic environment characterized by their inhabitants' high biological activity. Rhizospheric bacteria are recognized as effective biocontrol agents and considered cardinal in alternative strategies for securing ecological plant diseases management. The need to suppress fungal pathogens is an urgent task, not only because of the direct economic losses caused by infection but also due to their ability to produce mycotoxins with harmful effects on human health. Aspergillus and Fusarium species are well-known producers of toxigenic metabolites with a high capacity to colonize crops and enter the food chain. The bacteria belonging to the Bacillus genus has been conceded as a plant beneficial species in agricultural practice and identified as plant growth-promoting rhizobacteria (PGPR). Besides incontestable potential, the full commercialization of microbial biopesticides is in the preliminary phase. Thus, there is a constant need for estimating the suitability of novel strains to be used as a central point of viable bioprocess leading to market-ready product development. In the present study, 76 potential producing strains were isolated from the rhizosphere soil, sampled from different localities in the Autonomous Province of Vojvodina, Republic of Serbia. The selective isolation process of strains started by resuspending 1 g of soil samples in 9 ml of saline and incubating at 28° C for 15 minutes at 150 rpm. After homogenization, thermal treatment at 100° C for 7 minutes was performed. Dilution series (10-1-10-3) were prepared, and 500 µl of each was inoculated on nutrient agar plates and incubated at 28° C for 48 h. The pure cultures of morphologically different strains indicating belonging to the Bacillus genus were obtained by the spread-plate technique. The cultivation of the isolated strains was carried out in an Erlenmeyer flask for 96 h, at 28 °C, 170 rpm. The antagonistic activity screening included two phytopathogenic fungi as test microorganisms: Aspergillus sp. and Fusarium sp. The mycelial growth inhibition was estimated based on the antimicrobial activity testing of cultivation broth by the diffusion method. For the Aspergillus sp., the highest antifungal activity was recorded for the isolates Kro-4a and Mah-1a. In contrast, for the Fusarium sp., following 15 isolates exhibited the highest antagonistic effect Par-1, Par-2, Par-3, Par-4, Kup-4, Paš-1b, Pap-3, Kro-2, Kro-3a, Kro-3b, Kra-1a, Kra-1b, Šar-1, Šar-2b and Šar-4. One-way ANOVA was performed to determine the antagonists' effect statistical significance on inhibition zone diameter. Duncan's multiple range test was conducted to define homogenous groups of antagonists with the same level of statistical significance regarding their effect on antimicrobial activity of the tested cultivation broth against tested pathogens. The study results have pointed out the significant in vitro potential of the isolated strains to be used as biocontrol agents for the suppression of the tested mycotoxigenic fungi. Further research should include the identification and detailed characterization of the most promising isolates and mode of action of the selected strains as biocontrol agents. The following research should also involve bioprocess optimization steps to fully reach the selected strains' potential as microbial biopesticides and design cost-effective biotechnological production.

Keywords: Bacillus, biocontrol, bioprocess, mycotoxigenic fungi

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Authors: Fouzia Qamar, Shahida Hasnain

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The objective of the present study was to evaluate the possible role of Agrobacterium tumefaciens as a possible insect biocontrol agent. Pests selected for the present challenge were adult males of Periplaneta americana and last instar larvae of Pieris brassicae and Spodoptera litura. Different ranges of bacterial doses were selected and tested to score the mortalities of the insects after 24 hours, for the lethal dose estimation studies. Mode of application for the inoculation of the bacteria, was the microinjection technique. The evaluation of the possible entomopathogenic carrying attribute of bacterial Ti plasmid, led to the conclusion that the loss of plasmid was associated with the loss of virulence against target insects.

Keywords: agrobacterium tumefaciens, toxicity assessment, biopesticidal attribute, entomopathogenic agent

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Authors: Zuzana Sedrlova, Eva Slaninova, Ines Fritz, Christina Daffert, Stanislav Obruca

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Cyanobacteria are ecologically extremely important phototrophic gram-negative bacteria capable of oxygenic photosynthesis. They synthesize many interesting metabolites such as glycogen, carotenoids, but the most interesting metabolites are polyhydroxyalkanoates (PHA). The main advantage of cyanobacteria is the fact they do not require costly organic substrate and, oppositely, cyanobacteria can fix CO₂. PHA serves primarily as a carbon and energy source and occurs in the form of intracellular granules in bacterial cells. It is possible, PHA helps cyanobacteria to survive stress conditions since increased PHA synthesis was observed during cultivation in stress conditions. PHA is microbial biopolymers that are biodegradable with similar properties as petrochemical synthetic plastics. Production of PHA by heterotrophic bacteria is expensive; for price reduction waste materials as input, materials are used. Positively, cyanobacteria principally do not require organic carbon substrate since they are capable of CO₂ fixation. In this work, we demonstrated that stress conditions lead to the highest obtained yields of PHA in cyanobacterial cultures. Two cyanobacterial cultures from genera Synechocystis were used in this work. Cultivations were performed either in Erlenmayer flask or in tube multicultivator. Multiple stressors were applied on cyanobacterial cultures, and stressors include PHA precursors. PHA precursors are chemical substances and some of them do not occur naturally in the environment. Cultivation with the same PHA precursors in the same concentration led to a 1,6x higher amount of PHA when a multicultivator was used. The highest amount of PHA reached 25 % of PHA in dry cyanobacterial biomass. Both strains are capable of co-polymer synthesis in the presence of their structural precursor. The composition of co-polymer differs in Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. Synechocystis sp. PCC 6803 cultivated with γ-butyrolakton accumulated co-polymer of 3-hydroxybutyrate (3HB) and 4-hydroxybutyrate (4HB) the composition of the copolymer was 56 % of 4HB and 44 % of 3HB. The total amount of PHA, as well as yield of biomass, was lower than in control due to the toxic properties of γ-butyrolakton. Funding: This study was partly funded by the project GA19- 19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), a project I 4082-B25. This work was supported by Brno, Ph.D. Talent – Funded by the Brno City Municipality.

Keywords: co-polymer, cyanobacteria, PHA, synechocystis

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Authors: J. O. Alabi

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Biogas is a clean burning, easily produced natural fuel that is an important source of energy for cooking and heating in rural areas and third world countries. Anaerobic bacteria inside biodigesters break down biomass to produce biogas. (Which is 70% methane)? Currently there is no simple way to compress and store biogas. So, in order to use biogas as a source of energy, a direct feed from biodigeser to the store tap or heater must be made. Any excess biogas is vented into the atmosphere, which is wasteful and car have a negative effect on the environment, we have been tasked with designing a system that will be able to compress biogas using an off-grid power supply, making the biogas portable and makes through the use of large-scale, shared biodigester. Our final design is a system that maximizes simplicity and safety while minimizing cost.

Keywords: biogas, biodigesters, natural fuel, bionanotechnology

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Authors: Mokgadi Miranda Hlongwane, Ntebogeng Sharon Mokgalaka, Felix Dapare Dakora

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Lessertia (L.) frutescens (syn. Sutherlandia frutescens) is a leguminous medicinal plant indigenous to South Africa. Traditionally, L. frutescens has been used to treat cancer, diabetes, epilepsy, fever, HIV, stomach problems, wounds and other ailments. This legume is endemic to the Cape fynbos, with large populations occurring wild and cultivated in the Cape Florist Region. Its widespread distribution in the Western Cape, Northern Cape, Eastern Cape and Kwazulu-Natal is linked to its increased use as a phytomedicine in the treatment of various diseases by traditional healers. The frequent harvesting of field plants for use as a medicine has made it necessary to undertake studies towards the conservation of Lessertia frutescens. As a legume, this species can form root nodules and fix atmospheric N₂ when in symbiosis with soil bacteria called rhizobia. So far, however, few studies (if any) have been done on the efficacy and diversity of native bacterial symbionts nodulating L. frutescens in South Africa. The aim of this project was to isolate and characterize L. frutescens-nodulating bacteria from five different locations in the Western Cape Province. This was done by trapping soil rhizobia using rhizosphere soil suspension to inoculate L. frutescens seedlings growing in sterilized sand and receiving sterile N-free Hoagland nutrient solution under glasshouse conditions. At 60 days after planting, root nodules were harvested from L. frutescens plants, surface-sterilized, macerated, and streaked on yeast mannitol agar (YMA) plates and incubated at 28 ˚C for observation of bacterial growth. The majority of isolates were slow-growers that took 6-14 days to appear on YMA plates. However, seven isolates were fast-growers, taking 2-4 days to appear on YMA plates. Single-colony cultures of the isolates were assessed for their ability to nodulate L. frutescens as a homologous host under glasshouse conditions. Of the 92 bacterial isolates tested, 63 elicited nodule formation on L. frutescens. Symbiotic effectiveness varied markedly between and among test isolates. There were also significant (p≤0.005) differences in nodulation, shoot biomass, photosynthetic rates, leaf transpiration and stomatal conductance of L. frutescens plants inoculated with the test isolates, which is an indication of their functional diversity.

Keywords: lessertia frutescens, nodulating, rhizobia, symbiotic effectiveness

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Authors: F. A. Abdel-Mohdy, M. K. El-Bisi, A. Abou-Okeil, A. A. Sleem, S. El-Sabbagh, Kawther El-Shafei, Hoda S. El-Sayed, S. M. ElSawy

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Chitosan/ polyacrylic acid membranes containing different amounts of Al2(SO4) and/or TiO2 were prepared. The prepared membranes were characterized by measuring mechanical properties, such as tensile strength and elongation at break, swelling properties, antimicrobial properties against gram-positive and gram-negative bacteria and blood clotting. The results obtained indicate that the presence of Al2(SO4) and TiO2 in the membrane formulations have an incremental effect on the antimicrobial properties and blood clotting in albino rate.

Keywords: Chitosan, acrylic acid, antibacterial, blood clotting, membrane

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Authors: Karen Louise Lacey, Dario Javier Pavon Vargas, Massimiliano Rinaldi, Luca Cattani, Sara Rainieri

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The effects of high-pressure processing (HPP) and thermal treatments (TT) on quality markers of strawberry nectar (12 °Brix, 3,3 pH) was studied before and after treatments. TT and HPP treatments ensured a 3-log aerobic bacteria inactivation. No significant difference was detected in terms of pH and °Brix. TT samples were less red (a* less positive) than all HPP treated samples, while all samples were less red than the control. Apparent viscosity was significantly increased in all the HPP treatments, at 10 1/s shear rate, control was 79.04±7.94 mPa•s and the 600 MPa-20 min treatment were 327.10±1.64 mPa•s. This work suggests that HPP treatments may maintain the quality markers of strawberry nectar better.

Keywords: HPP, strawberry nectar, colour , viscosity

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Authors: Elmo Thiago Lins Cöuras Ford, Valentina Alessandra Carvalho do Vale

Abstract:

It presents a heating system using low cost alternative solar collectors to promote the disinfection of water in low income communities that take water contaminated by bacteria. The system consists of two solar collectors, with total area of 4 m² and was built using PET bottles and cans of beer and soft drinks. Each collector is made up of 8 PVC tubes, connected in series and work in continuous flow. It will determine the flux the most appropriate to generate the temperature to promote the disinfection. It will be presented results of the efficiency and thermal loss of system and results of analysis of water after undergoing the process of heating.

Keywords: Disinfection of water, solar heating system, poor communities, bioinformatics, biomedicine

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Authors: Abdessemed Zineb, Atia Yahia, Yeza Salima

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Water is self-purified by activated sludge process which makes its uniqueness. The main goal is the microbial biocenosis study of the input and output water of the waste water treatment system plant Timgad. 89.47% of the identified biocenosis belongs to ɤ-Proteobacteria while the remaining 10.52 % is equally divided between α-Proteobacteria and β-Proteobacteria. The antibiotics susceptibility profiles reveal that over 30 % are wild strains while the penicillinases are often present (11.30-20 %) with also other profiles. This proportion is worrying that the water discharged join the Oued Soltez used for irrigation. This disadvantage involves the installation of a chlorination step.

Keywords: activated sludge, biocenosis, antibiotics profiles, penicillinases, physic-chemical quality

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Authors: Tariku Nefo Duke

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Embelia is a genus of climbing shrubs in the family Myrsinaceae. Embelia schimperi is as important in traditional medicine as the other species in the genus. The plant has been much known as a local medicine for the treatment of tapeworms. In this project, extraction, phytochemical screening tests, isolation, and characterization of berries of the Embelia schimperi plant have been conducted. The chemical investigations of methanol and ethyl acetate (1:1) ratio extracts of the berries lead to the isolation of three new compounds. The compounds were identified to be alkaloids coded as AD, AN, and AG. Structural elucidations of the isolated compounds were accomplished using spectroscopic methods (IR, UV, ¹H NMR, ¹³C NMR, DEPT and 2D NMR, HPLC, and LC-MS). The alkaloid coded as (AN) has a wide MIC range of 6.31-25.46 mg/mL against all tested bacteria strains.

Keywords: Embelia schimper, HPLC, alkaloids, 2D NMR, MIC

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Authors: Zexiao Zheng, Irene M. C. Lo

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Organic pollutants, ammonia, and bacteria are major contaminants in sewage, which may adversely impact ecosystems without proper treatment. Conventional wastewater treatment plants (WWTPs) are operated to remove these contaminants from sewage but suffer from high carbon emissions and are powerless to remove emerging organic pollutants (EOPs). Herein, we have developed a low operational carbon emissions multifunctional photoelectrochemical (PEC) system for saline sewage treatment to simultaneously remove organic compounds, ammonia, and bacteria, coupled with H2 evolution. A reduced BiVO4 (r-BiVO4) with improved PEC properties due to the construction of oxygen vacancies and V4+ species was developed for the multifunctional PEC system. The PEC/r-BiVO4 process could treat saline sewage to meet local WWTPs’ discharge standard in 40 minutes at 2.0 V vs. Ag/AgCl and completely degrade carbamazepine (one of the EOPs), coupled with significant evolution of H2. A remarkable reduction in operational carbon emissions was achieved by the PEC/r-BiVO4 process compared with large-scale WWTPs, attributed to the restrained direct carbon emissions from the generation of greenhouse gases. Mechanistic investigation revealed that the PEC system could activate chloride ions in sewage to generate reactive chlorine species and facilitate •OH production, promoting contaminants removal. The PEC system exhibited operational feasibility at different pH and total suspended solids concentrations and has outstanding reusability and stability, confirming its promising practical potential. The study combined the simultaneous removal of three major contaminants from saline sewage and H2 evolution in a single PEC process, demonstrating a viable approach to supplementing and extending the existing wastewater treatment technologies. The study generated profound insights into the in-situ activation of existing chloride ions in sewage for contaminants removal and offered fundamental theories for applying the PEC system in sewage remediation with low operational carbon emissions. The developed PEC system can fit well with the future needs of wastewater treatment because of the following features: (i) low operational carbon emissions, benefiting the carbon neutrality process; (ii) higher quality of the effluent due to the elimination of EOPs; (iii) chemical-free in the operation of sewage treatment; (iv) easy reuse and recycling without secondary pollution.

Keywords: contaminants removal, H2 evolution, multifunctional PEC system, operational carbon emissions, saline sewage treatment, r-BiVO4 photoanodes

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Authors: Abdul Walusansa

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In this paper, paper notes and coins were collected from general public in Kampala City where ready-to-eat food can be served, in order to survey for bacterial contamination. The total bacterial number and potentially pathogenic organisms loading on currency were tested. All isolated potential pathogens were also tested for antibiotic resistance against four most commonly prescribed antibiotics. 1. The bacterial counts on one hundred paper notes sample were ranging between 6～10918/cm cm-2,the median was 141/ cm-2, according to the data it was much higher than credit cards and Australian notes which were made of polymer. The bacterial counts on sixty coin samples were ranging between 2～380/cm-2, much less than paper notes. 2. Coliform (65.6%), E. coli (45.9%), S. aureus (41.7%), B. cereus (67.7%), Salmonella (19.8%) were isolated on one hundred paper notes. Coliform (22.4%), E. coli (5.2%), S. aureus (24.1%), B. cereus (34.5%), Salmonella (10.3%) were isolated from sixty coin samples. These results suggested a high rate of potential pathogens contamination of paper notes than coins. 3. Antibiotic resistances are commonly in most of the pathogens isolated on currency. Ampicillin resistance was found in 60%of Staphylococcus aureus isolated on currency, as well as 76.6% of E. coil and 40% of Salmonella. Erythromycin resistance was detected in 56.6% of S. aureus and in 80.0% of E. coli. All the pathogens isolated were sensitive to Norfloxacin, Salmonella and S. aureus also sensitive to Cefaclor. In this paper, we also studied the antimicrobial capability of metal coins, coins collected from different countries were tested for the ability to inhibit the growth of E. sakazakii, S. aureus, E. coli, L. monocytogenes and S. typhimurium. 1) E. sakazakii appeared very sensitive to metal coins, the second is S. aureus, but E. coli, L. monocytogenes and S. typhimurium are more resistant to these metal coin samples. 2) Coins made of Nickel-brass alloy and Copper-nickel alloy showed a better effect in anti-microbe than other metal coins, especially the ability to inhibited the growth of E. sakazakii and S. aureus, all the inhibition zones produced on nutrient agar are more than 20.6 mm. Aluminium-bronze alloy revealed weak anti-microbe activity to S. aureus and no effect to kill other pathogens. Coins made of stainless steel also can’t resist bacteria growth. 3) Surprisingly, one cent coins of USA which were made of 97.5% Zinc and 2.5% Cu showed a significant antimicrobial capability, the average inhibition zone of these five pathogens is 45.5 mm.

Keywords: antibiotic sensitivity, bacteria, currency, coins, parasites

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Authors: S. Mamoucha, N.Tsafantakis, T. Ioannidis, S. Chatzipanagiotou, C. Nikolaou, L. Skaltsounis, N. Fokialakis, N. Christodoulakis

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Introduction: Pistacia lentiscus L. (well known as Mastic tree) is an evergreen sclerophyllous shrub that extensively thrives in the eastern Mediterranean area yet only the trees cultivated in the southern region of the Greek island Chios produces mastic resin. Different parts of P. lentiscus L. var. chia have been used in folk medicine for various purposes, such as tonic, aphrodisiac, antiseptic, antihypertensive and management of dental, gastrointestinal, liver, urinary, and respiratory tract disorders. Several studies have focused on the antibacterial activity of its resin (gum) and its essential oil. However, there is no study combining anatomy of the plant organs, phytochemical profile, and antibacterial screening of the plant. In our attempt to discover novel bioactive metabolites from the mastic tree, we screened its antibacterial activity not only against ATCC strains but also against clinical, resistant strains. Materials-methods: Leaves were investigated using Transmission (ΤΕΜ) and Scanning Εlectron Microscopy (SEM). Histochemical tests were performed on fresh and fixed tissue. Extracts prepared from dried, powdered leaves using 3 different solvents (DCM, MeOH and H2O) the waste water obtained after a hydrodistillation process for essential oil production were screened for their phytochemical content and antibacterial activity. Μetabolite profiling of polar and non-polar extracts was recorded by GC-MS and LC-HRMS techniques and analyzed using in-house and commercial libraries. The antibacterial screening was performed against Staphylococcus aureus ATCC25923, Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853 and against clinical, resistant strains Methicillin-resistant S. aureus (MRSA), Carbapenem-Resistant Metallo-β-Lactamase (carbapenemase) P. aeruginosa (VIM), Klebsiella pneumoniae carbapenemases (KPCs) and Acinetobacter baumanii resistant strains. The antibacterial activity was tested by the Kirby Bauer and the Agar Well Diffusion method. The zone of inhibition (ZI) of each extract was measured and compared with those of common antibiotics. Results: Leaf is compact with inosclereids and numerous idioblasts containing a globular, spiny crystal. The major nerves of the leaf contain a resin duct. Mesophyll cells showed accumulation of osmiophillic metabolites. Histochemical treatments defined secondary metabolites in subcellular localization. The phytochemical investigation revealed the presence of a large number of secondary metabolites, belonging to different chemical groups, such as terpenoids, phenolic compounds (mainly myricetin, kaempferol and quercetin glycosides), phenolic, and fatty acids. Among the extracts, the hydrostillation wastewater achieved the best results against most of the bacteria tested. MRSA, VIM and A. baumanii were inhibited. Conclusion: Extracts from plants have recently been of great interest with respect to their antimicrobial activity. Their use emerged from a growing tendency to replace synthetic antimicrobial agents with natural ones. Leaves of P. lentiscus L. var. chia showed a high antimicrobial activity even against drug - resistant bacteria. Future prospects concern the better understanding of mode of action of the antibacterial activity, the isolation of the most bioactive constituents and the clarification if the activity is related to a single compound or to the synergistic effect of several ones.

Keywords: antibacterial screening, leaf anatomy, phytochemical profile, Pistacia lentiscus var. chia

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Authors: Elif Tugce Aksun Tumerkan, Chris Lowe, Hannah Krupa

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Fluorescent proteins are self-sufficient in forming chromophores with a visible wavelength from 3 amino acids sequence within their own polypeptide structure. This chromophore – a molecule that absorbs a photon of light and exhibits an energy transition equal to the energy of the absorbed photon. Fluorescent proteins (FPs) consisted of a chain of 238 amino acid residues and composed of 11 beta strands shaped in a cylinder surrounding an alpha helix structure. A better understanding of the system of the chromospheres and the increasing advance in protein engineering in recent years, the properties of FPs offers the potential for new applications. They have used sensors and probes in molecular biology and cell-based research that giving a chance to observe these FPs tagged cell localization, structural variation and movement. For clarifying functional uses of fluorescent proteins, electrophoretic properties of these proteins are one of the most important parameters. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is used for determining electrophoretic properties commonly. While there are many techniques are used for determining the functionality of protein-based research, SDS-PAGE analysis can only provide a molecular level assessment of the proteolytic fragments. Before SDS-PAGE analysis, fluorescent proteins need to successfully purified. Due to directly purification of the target, FPs is difficult from the animal, gene expression is commonly used which must be done by transformation with the plasmid. Furthermore, used gel within electrophoresis and staining agents properties have a key role. In this review, the different factors that have the impact on the electrophoretic properties of fluorescent proteins explored. Fluorescent protein separation and purification are the essential steps before electrophoresis that should be done very carefully. For protein purification, gene expression process and following steps have a significant function. For successful gene expression, the properties of selected bacteria for expression, used plasmid are essential. Each bacteria has own characteristics which are very sensitive to gene expression, also used procedure is the important factor for fluorescent protein expression. Another important factors are gel formula and used staining agents. Gel formula has an effect on the specific proteins mobilization and staining with correct agents is a key step for visualization of electrophoretic bands of protein. Visuality of proteins can be changed depending on staining reagents. Apparently, this review has emphasized that gene expression and purification have a stronger effect than electrophoresis protocol and staining agents.

Keywords: cell biology, gene expression, staining agents, SDS-page

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Authors: Elvan G. Bulut, Betul Goksun, Tugba G. Gun, Ozge Sakiyan Demirkol, Kamuran Ayhan, Kezban Candogan

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Dysphagia, difficulty in swallowing solid foods and thin liquids, is one of the common health threats among the elderly who require foods with modified texture in their diet. Although there are some commercial food formulations or hydrocolloids to thicken the liquid foods for dysphagic individuals, there is still a need for developing and offering new food products with enriched nutritional, textural and sensory characteristics to safely nourish these patients. 3D food printing is an appealing alternative in creating personalized foods for this purpose with attractive shape, soft and homogenous texture. In order to modify texture and prevent phase separation, hydrocolloids are generally used. In our laboratory, an optimized 3D printed beef-based formulation specifically for people with swallowing difficulties was developed based on the research project supported by the Scientific and Technological Research Council of Turkey (TÜBİTAK Project # 218O017). The optimized formulation obtained from response surface methodology was 60% beef powder, 5.88% gelatin, and 0.74% kappa-carrageenan (all in a dry basis). This product was enriched with powders of freeze-dried beet, celery, and red capia pepper, butter, and whole milk. Proximate composition (moisture, fat, protein, and ash contents), pH value, CIE lightness (L*), redness (a*) and yellowness (b*), and color difference (ΔE*) values were determined. Counts of total mesophilic aerobic bacteria (TMAB), lactic acid bacteria (LAB), mold and yeast, total coliforms were conducted, and detection of coagulase positive S. aureus, E. coli, and Salmonella spp. were performed. The 3D printed products had 60.11% moisture, 16.51% fat, 13.68% protein, and 1.65% ash, and the pH value was 6.19, whereas the ΔE* value was 3.04. Counts of TMAB, LAB, mold and yeast and total coliforms before and after 3D printing were 5.23-5.41 log cfu/g, < 1 log cfu/g, < 1 log cfu/g, 2.39-2.15 log EMS/g, respectively. Coagulase positive S. aureus, E. coli, and Salmonella spp. were not detected in the products. The data obtained from this study based on determining some important product characteristics of functional beef-based formulation provides an encouraging basis for future research on the subject and should be useful in designing mass production of 3D printed products of similar composition.

Keywords: beef, dysphagia, product characteristics, texture-modified foods, 3D food printing

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Authors: Astrid Tannert, Anuradha Ramoji, Christina Ebert, Frederike Gladigau, Lorena Tuchscherr, Jürgen Popp, Ute Neugebauer

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Staphylococcus aureus is a facultative intracellular pathogen, which by entering host cells may evade immunologic host response as well as antimicrobial treatment. In that way, S. aureus can cause persistent intracellular infections which are difficult to treat. Depending on the strain, S. aureus may persist at different intracellular locations like the phagolysosome. The first barrier invading pathogens from the blood stream that they have to cross are the endothelial cells lining the inner surface of blood and lymphatic vessels. Upon proceeding from an acute to a chronic infection, intracellular pathogens undergo certain biochemical and structural changes including a deceleration of metabolic processes to adopt for long-term intracellular survival and the development of a special phenotype designated as small colony variant. In this study, the endothelial cell line Ea.hy 926 was used as a model for acute and chronic S. aureus infection. To this end, Ea.hy 926 cells were cultured on QIAscout™ Microraft Arrays, a special graded cell culture substrate that contains around 12,000 microrafts of 200 µm edge length. After attachment to the substrate, the endothelial cells were infected with GFP-expressing S. aureus for 3 weeks. The acute infection and the development of persistent bacteria was followed by confocal laser scanning microscopy, scanning the whole Microraft Array for the presence and for detailed determination of the intracellular location of fluorescent intracellular bacteria every second day. After three weeks of infection representative microrafts containing infected cells, cells with protruded infections and cells that did never show any infection were isolated and fixed for Raman micro-spectroscopic investigation. For comparison, also microrafts with acute infection were isolated. The acquired Raman spectra are correlated with the fluorescence microscopic images to give hints about a) the molecular alterations in endothelial cells during acute and chronic infection compared to non-infected cells, and b) metabolic and structural changes within the pathogen when entering a mode of persistence within host cells. We thank Dr. Ruth Kläver from QIAGEN GmbH for her support regarding QIAscout technology. Financial support by the BMBF via the CSCC (FKZ 01EO1502) and from the DFG via the Jena Biophotonic and Imaging Laboratory (JBIL, FKZ PO 633/29-1, BA 1601/10-1) is highly acknowledged.

Keywords: correlative image analysis, intracellular infection, pathogen-host adaption, Raman micro-spectroscopy

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Authors: Frans J. Smit, Wisdom A. Munzeiwa, Hermanus C. M. Vosloo, Lyn-Marie Birkholtz, Richard K. Haynes

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Malaria and antimicrobial infections remain significant global health concerns, necessitating the continuous search for novel therapeutic approaches. This abstract presents an overview of the potential use of triazenes as effective agents against malaria and various antimicrobial pathogens. Triazenes are a class of compounds characterized by a linear arrangement of three nitrogen atoms, rendering them structurally distinct from their cyclic counterparts. This study investigates the efficacy of triazenes against malaria and explores their antimicrobial activity. Preliminary results revealed significant antimalarial activity of the triazenes, as evidenced by in vitro screening against P. falciparum, the causative agent of malaria. Furthermore, the compounds exhibited broad-spectrum antimicrobial activity, indicating their potential as effective antimicrobial agents. These compounds have shown inhibitory effects on various essential enzymes and processes involved in parasite survival, replication, and transmission. The mechanism of action of triazenes against malaria involves interactions with critical molecular targets, such as enzymes involved in the parasite's metabolic pathways and proteins responsible for host cell invasion. The antimicrobial activity of the triazenes against bacteria and fungi was investigated through disc diffusion screening. The antimicrobial efficacy of triazenes has been observed against both Gram-positive and Gram-negative bacteria, as well as multidrug-resistant strains, making them potential candidates for combating drug-resistant infections. Furthermore, triazenes possess favourable physicochemical properties, such as good stability, solubility, and low toxicity, which are essential for drug development. The structural versatility of triazenes allows for the modification of their chemical composition to enhance their potency, selectivity, and pharmacokinetic properties. These modifications can be tailored to target specific pathogens, increasing the potential for personalized treatment strategies. In conclusion, this study highlights the potential of triazenes as promising candidates for the development of novel antimalarial and antimicrobial therapeutics. Further investigations are necessary to determine the structure-activity relationships and optimize the pharmacological properties of these compounds. The results warrant additional research, including MIC studies, to further explore the antimicrobial activity of the triazenes. Ultimately, these findings contribute to the development of more effective strategies for combating malaria and microbial infections.

Keywords: malaria, anti-microbials, triazene, resistance

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Authors: Andrea M. Rivas-Castillo, Marlenne Gómez-Ramirez, Isela Rodríguez-Pozos, Norma G. Rojas-Avelizapa

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Spent catalysts are considered as hazardous residues of major concern, mainly due to the simultaneous presence of several metals in elevated concentrations. Although hydrometallurgical, pyrometallurgical and chelating agent methods are available to remove and recover some metals contained in spent catalysts; these procedures generate potentially hazardous wastes and the emission of harmful gases. Thus, biotechnological treatments are currently gaining importance to avoid the negative impacts of chemical technologies. To this end, diverse microorganisms have been used to assess the removal of metals from spent catalysts, comprising bacteria, archaea and fungi, whose resistance and metal uptake capabilities differ depending on the microorganism tested. Acidophilic sulfur oxidizing bacteria have been used to investigate the biotreatment and extraction of valuable metals from spent catalysts, namely Acidithiobacillus thiooxidans and Acidithiobacillus ferroxidans, as they present the ability to produce leaching agents such as sulfuric acid and sulfur oxidation intermediates. In the present work, the ability of A. thiooxidans DSM 26636 for the bioleaching of metals contained in five different spent catalysts was assessed by growing the culture in modified Starkey mineral medium (with elemental sulfur at 1%, w/v), and 1% (w/v) pulp density of each residue for up to 21 days at 30 °C and 150 rpm. Sulfur-oxidizing activity was periodically evaluated by determining sulfate concentration in the supernatants according to the NMX-k-436-1977 method. The production of sulfuric acid was assessed in the supernatants as well, by a titration procedure using NaOH 0.5 M with bromothymol blue as acid-base indicator, and by measuring pH using a digital potentiometer. On the other hand, Inductively Coupled Plasma - Optical Emission Spectrometry was used to analyze metal removal from the five different spent catalysts by A. thiooxidans DSM 26636. Results obtained show that, as could be expected, sulfuric acid production is directly related to the diminish of pH, and also to highest metal removal efficiencies. It was observed that Al and Fe are recurrently removed from refinery spent catalysts regardless of their origin and previous usage, although these removals may vary from 9.5 ± 2.2 to 439 ± 3.9 mg/kg for Al, and from 7.13 ± 0.31 to 368.4 ± 47.8 mg/kg for Fe, depending on the spent catalyst proven. Besides, bioleaching of metals like Mg, Ni, and Si was also obtained from automotive spent catalysts, which removals were of up to 66 ± 2.2, 6.2±0.07, and 100±2.4, respectively. Hence, the data presented here exhibit the potential of A. thiooxidans DSM 26636 for the simultaneous bioleaching of metals contained in spent catalysts from diverse provenance.

Keywords: bioleaching, metal removal, spent catalysts, Acidithiobacillus thiooxidans

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Authors: Magdalena Diaka, Paweł Mazierski, Joanna Żebrowska, Michał Winiarski, Tomasz Klimczuk, Adriana Zaleska-Medynska

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During the last years, TiO2 nanotubes have been widely studied due to their unique highly ordered array structure, unidirectional charge transfer and higher specific surface area compared to conventional TiO2 powder. These photoactive materials, in the form of thin layer, can be activated by low powered and low cost irradiation sources (such as LEDs) to remove VOCs, microorganism and to deodorize air streams. This is possible due to their directly growth on a support material and high surface area, which guarantee enhanced photon absorption together with an extensive adsorption of reactant molecules on the photocatalyst surface. TiO2 nanotubes exhibit also lots of other attractive properties, such as potential enhancement of electron percolation pathways, light conversion, and ion diffusion at the semiconductor-electrolyte interface. Pure TiO2 nanotubes were previously used to remove organic compounds from the gas phase as well as in water splitting reaction. The major factors limiting the use of TiO2 nanotubes, which have not been fully overcome, are their relatively large band gap (3-3,2 eV) and high recombination rate of photogenerated electron–hole pairs. Many different strategies were proposed to solve this problem, however titania nanostructures containing incorporated metal oxides like Ag2O shows very promising, new optical and photocatalytic properties. Unfortunately, there is still very limited number of reports regarding application of TiO2/MxOy nanostructures. In the present work, we prepared TiO2/Ag2O nanotubes obtained by anodization of Ti-Ag alloys containing 5, 10 and 15 wt. % Ag. Photocatalysts prepared in this way were characterized by X-ray diffraction spectroscopy (XRD), scanning electron microscopy (SEM), luminescence spectroscopy and UV-Vis spectroscopy. The activities of new TiO2/Ag2O were examined by photocatalytic degradation of toluene in gas phase reaction and phenol in aqueous phase using 1000 W Xenon lamp (Oriel) and light emitting diodes (LED) as a irradiation sources. Additionally efficiency of bacteria (Pseudomonas aeruginosa) removal from the gas phase was estimated. The number of surviving bacteria was determined by the serial twofold dilution microtiter plate method, in Tryptic Soy Broth medium (TSB, GibcoBRL).

Keywords: photocatalysis, antibacterial properties, titania nanotubes, new TiO2/MxOy nanostructures

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Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash

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Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.

Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking

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