The study of biofilm function, structure and microbial interactions might help to improve our understanding of biofilm wastewater treatment processes. However, few reports specifically address the influence of interactions within multispecies biofilms on microbial activity and biofilm composition. Thus, the relationship between biofilm formation, denitrification activity, phosphorus removal and the composition of extracellular polymeric substances (EPS), exopolysaccharides and the bacterial community was investigated using biofilms of denitrifying and phosphorus removing strains Comamonas denitrificans 110, Brachymonas denitrificans B79, Aeromonas hydrophila L6 and Acinetobacter calcoaceticus ATCC23055. Denitrification activity within the biofilms generally increased with the amount of biofilm while phosphorus removal depended on bacterial growth rate. Synergistic effects of co-growth on denitrification (B. denitrificans B79 and A. hydrophila L6) and phosphorus removal (C. denitrificans 110 with either A. calcoaceticus or A. hydrophila L6) were observed. B. denitrificans B79 was highly affected by interspecies interactions with respect to biofilm formation, denitrification activity and EPS composition, while C. denitrificans 110 remained largely unaffected. In some of the dual and quadruple strain biofilms new exopolysaccharide monomers were detected which were not present in the pure strain samples.
Extracellular polymeric substances (EPS) play an important role in the formation and activity of biofilms in wastewater treatment (WWT). The EPS of the denitrifying biomarker Comamonas denitrificans strain 110, produced in different culture media and growth modes, were characterized. The EPS mainly contained protein (3-37%), nucleic acids (9-50%), and carbohydrates (3-21%). The extracellular DNA was found to be important for initial biofilm formation since biofilm, but not planktonic growth, was inhibited in the presence of DNase. The polysaccharide fraction appeared to consist of at least two distinct polymers, one branched fraction (A) made up of glucose and mannose with a molecular weight around 100 kDa. The other fraction (B) was larger and consisted of ribose, mannose, glucose, rhamnose, arabinose, galactose, and N-acetylglucosamine. Fraction B polysaccharides were mainly found in capsular EPS which was the dominant type in biofilms and agar-grown colonies. Fraction A was abundant in the released EPS, the dominant type in planktonic cultures. Biofilm and agar-grown EPS displayed similar overall properties while planktonic EPS showed clear compositional disparity. This study presents results on the physiology of a key WWT organism, which may be useful in the future development of improved biofilm techniques for WWT purposes.
Biofilm formation and adherence properties of 13 bacterial strains commonly found in wastewater treatment systems were studied in pure and mixed cultures using a crystal violet microtiter plate assay. Four different culture media were used, wastewater, acetate medium, glucose medium and diluted nutrient broth. The medium composition strongly affected biofilm formation. All strains were able to form pure culture biofilms within 24 h in at least one of the tested culture media and three strains were able to form biofilm in all four culture media, namely Acinetobacter calcoaceticus ATCC 23055, Comamonas denitrificans 123 and Pseudomonas aeruginosa MBL 0199. The adherence properties assessed were initial adherence, cell surface hydrophobicity, and production of amyloid fibers and extracellular polymeric substances. The growth of dual-strain biofilms showed that five organisms formed biofilm with all 13 strains while seven formed no or only weak biofilm when cocultured. In dual-strain cultures, strains with different properties were able to complement each other, giving synergistic effects. Strongest biofilm formation was observed when a mixture of all 13 bacteria were grown together. These results on attachment and biofilm formation can serve as a tool for the design of tailored systems for the degradation of municipal and industrial wastewater.
The pine weevil, Hylobius abietis, is a severe forest pest insect as it feeds on newly planted conifer seedlings. To identify and develop an antifeedant could be one step towards the protection of seedlings from feeding damage by the pine weevil. With the aim to trace the origin of the antifeedants previously found in feces of the pine weevil, we investigated the culturable bacteria associated with the gut and identified the volatiles they produced. Bacterial isolates were identified by 16S ribosomal RNA gene analysis. The volatile emissions of selected bacteria, cultivated on NB media or on the grated phloem of Scots pine twigs dispersed in water, were collected and analyzed by solid-phase microextraction gas chromatography-mass spectrometry. The bacterial isolates released a variety of compounds, among others 2-methoxyphenol, 2-phenylethanol, 3-methyl-1-butanol, 1-octen-3-ol, 3-octanone, dimethyl disulfide, and dimethyl trisulfide. A strong antifeedant effect was observed by 2-phenylethanol, which could thus be a good candidate for use to protect planted conifer seedlings against feeding damage caused by H. abietis.
Heterobasidion annosum s.s. and H. parviporum are severe pathogens of conifers causing butt rot and root rot thus reducing the economic value of timber. Here, the antifungal activity of Bacillus subtilis isolate A18 against these two Heterobasidion species was investigated. Five different culture media with different culture age were investigated to study the effect of substrate composition and culture age for metabolite production. Bacterial cultures and cell-free culture filtrates were tested for antifungal activity. Inhibition of fungal growth was analysed using the agar disc-diffusion method. MALDI-TOF and LC-HRMS analyses were used to identify the antifungal metabolites. Substrate composition and age of culture were found to be active variables with direct effect on the antifungal activity of bacterial culture extracts. High anti-fungal activity was observed when B. subtilis was cultured in PDB, SGB and LB media for four days. Mass-spectrometry analysis showed the presence of lipopeptides in culture filtrates identified as members of the surfactins, polymixins, kurstakins and fengycins. A culture filtrate containing fengycin-type lipopeptides showed the highest bioactivity against Heterobasidion species. Bacterial cultures had higher bioactivity compared to their respective cell free culture filtrates. The results of the present study suggest that B. subtilis A18 is a powerful biocontrol agent against Heterobasidion infections of tree wounds and stumps.
A strain of Penicillium expansum was studied for the production of styrene using forest waste biomass as a feeding substrate. The fungal strain was cultivated on bark of various trees supplemented with yeast extract and the volatiles produced were collected on Tenax TA and analyzed by gas chromatography-mass spectrometry. Fungus cultured on grated soft bark of pine (Pinus sylvestris) stems (GPB) and mature bark of oak (Quercus robur) supplemented with yeast extract produced relatively the highest amounts of styrene. The maximum styrene production rate was 52.5 mu g/h, 41 mu g/h and 27 mu g/h from fungus cultivated on 50 mL liquid media with 10 g GPB or mature bark of oak and potato dextrose broth respectively. These promising results suggest that the fungal strain could be used to produce "green" styrene plastics using renewable forest waste biomass.
The pine weevil Hylobius abietis (L.) is a severe pest of conifer seedlings in reforested areas of Europe and Asia. To identify minimally toxic and ecologically sustainable compounds for protecting newly planted seedlings, we evaluated the volatile metabolites produced by microbes isolated from H. abietis feces and frass. Female weevils deposit feces and chew bark at oviposition sites, presumably thus protecting eggs from feeding conspecifics. We hypothesize that microbes present in feces/frass are responsible for producing compounds that deter weevils. Here, we describe the isolation of a fungus from feces and frass of H. abietis and the biological activity of its volatile metabolites. The fungus was identified by morphological and molecular methods as Penicillium expansum Link ex. Thom. It was cultured on sterilized H. abietis frass medium in glass flasks, and volatiles were collected by SPME and analyzed by GC-MS. The major volatiles of the fungus were styrene and 3-methylanisole. The nutrient conditions for maximum production of styrene and 3-methylanisole were examined. Large quantities of styrene were produced when the fungus was cultured on grated pine bark with yeast extract. In a multi-choice arena test, styrene significantly reduced male and female pine weevils' attraction to cut pieces of Scots pine twigs, whereas 3-methylanisole only reduced male weevil attraction to pine twigs. These studies suggest that metabolites produced by microbes may be useful as compounds for controlling insects, and could serve as sustainable alternatives to synthetic insecticides.
The pine weevil is one of the most important pest insects of conifer reforestation areas in Europe. Female pine weevils cover their eggs with chewed bark and feces (frass) resulting in avoidance behavior of feeding conspecifics towards egg laying sites. It has been suggested that microorganisms present in the frass may be responsible for producing deterrent compounds for the pine weevil. The fungi Ophiostoma canum, O. pluriannulatum, and yeast Debaryomyces hansenii were isolated from aseptically collected pine-weevil frass. The isolated fungi were cultured on weevil frass broth and their volatiles were collected by SPME and identified by GC MS. D. hansenii produced methyl salicylate (MeS) as a major compound, whereas, in addition, O. canum and O. pluriannulatum produced 6-protoilludene. In a multi-choice lab bioassay, MeS strongly reduced pine weevil's attraction to the Pinus sylvestris volatiles. Thus, a fungal metabolite was found that strongly affects the pine weevil host-odor search. (C) 2014 Elsevier Ltd and The British Mycological Society. All rights reserved.
The pine weevil Hylobius abietis is a severe pest of conifer seedlings in reforestation areas. Weevils lay eggs in the root bark or in the soil near roots of recently dead trees and cover the eggs with frass (feces combined with chewed bark), possibly to avoid conspecific egg predation. The aim of the present investigation focused on isolation, identification, and volatile production of fungi from pine-weevil feces and frass. Fungi were isolated from weevil frass and feces separately, followed by identification based on ITS sequencing. Fifty-nine isolates belonging to the genera Penicillium, Ophiostoma, Mucor, Leptographium, Eucasphaeria, Rhizosphaera, Debaryomyces, and Candida were identified. Volatile organic compounds (VOCs) produced by the fungal community and fungal isolates cultured on weevil-frass broth were identified by SPME-GCMS. Major VOCs emitted from the fungal community and pure isolates were species- and strain specific and included isopentylalcohol, styrene, 3-octanone, 6-protoilludene, methyl salicylate, 3-methylanisole, 2-methoxyphenol, and phenol. Some of these are known to influence the orientation of pine weevils when tested among highly attractive newly planted conifer seedlings.
Functionalization of super paramagnetic iron oxide NPs (SPIONs) with different coatings renders them with unique physicochemical properties that allow them to be used in a broad range of applications such as drug targeting and water purification. However, it is required to fill the gap between the promises of any new functionalized SPIONs and the effects of these coatings on the NPs safety. Nanotoxicology is offering diverse strategies to assess the effect of exposure to SPIONs in a case-by-case manner but an integrated nanoimpact scale has not been developed yet. We have implemented the classical integrated biological response (IBR) into an integrated nanoimpact index (INI) as an early warning scale of nano-impact based on a combination of toxicological end points such as cell proliferation, oxidative stress, apoptosis and genotoxicity. Here, the effect of SPIONs functionalized with tri-sodium citrate (TSC), polyethylenimine (PEI), aminopropyl-triethoxysilane (APTES) and Chitosan (chitosan) were assessed on human keratinocytes and endothelial cells. Our results show that endothelial cells were more sensitive to exposure than keratinocytes and the initial cell culture density modulated the toxicity. PEI-SPIONs had the strongest effects in both cell types while TSC-SPIONS were the most biocompatible. This study emphasizes not only the importance of surface coatings but also the cell type and the initial cell density on the selection of toxicity assays. The INI developed here could offer an initial rationale to choose either modifying SPIONs properties to reduce its nanoimpact or performing a complete risk assessment to define the risk boundaries.
The use of natural coagulant protein in drinking water treatment has been discussed for a long time, though the method is still not in practice, probably due to limited knowledge and availability of material. In the present work, different Mustard varieties were tested for the presence of coagulant protein compared with Moringa seed extract and their potential application in water treatment. The coagulation activity of the protein extract was measured using synthetic clay solution as well as water from pond. The protein content was determined by Bradford method, molecular mass determined by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and peptide sequence was analyzed by Mass spectrometry. Extract of Mustard (large) and Moringa seed showed coagulation activity of a parts per thousand...70 and a parts per thousand...85 % after 90 min, respectively. Interestingly, seed extracts from other Mustard varieties had coagulation activity after heat activation at 95 A degrees C for 5 h. However, the coagulation activity of Mustard seed extract against turbid pond water was higher (a parts per thousand...60 %) compared to Moringa seed extract (a parts per thousand...50 %). The peptide sequence analysis of 6.5 and 9 kDa proteins was found to be homologous to Moringa coagulant protein and napin3, respectively. To our knowledge, this could be the first report on Mustard seed having coagulant protein. The coagulation activity of Mustard (large) against highly turbid pond water suggested that it could be a potential natural coagulant for water treatment.
Access to fresh water is limited due to the growing population and to provide safe drinking water is a global challenge. In this initial study, about a hundred different samples were screened for coagulation activity. Amongst the plants tested were fruits and vegetable pulps, leaves and seeds (like jackfruit, tamarind, papaya, orange, watermelon, pineapple, cucumber, green gram, peas, black gram, cluster bean rice, maize, chili, cabbage, mustard, cauliflower, beetroot and carrot). The crude extracts were prepared in water and salt solution in order to extract the coagulant protein and compare the coagulation activity. Among the tested samples, only a few seed extracts showed coagulation activity. The seed extracts of mustard and cabbage showed coagulation activity in both water and salt extracts and were comparable to that of Moringa seed extract. The salt extracts showed higher activity compared to that of water extract in most of the samples tested. The coagulation activity was stable after incubation at 95 WC for 5 hours implying heat resistance. A protein profile analysis showed major protein bands with a molecular weight around 6.5 and 9 kDa. In this preliminary study, mustard and cabbage seed extracts were promising in terms of coagulation and heat resistance, so could be an alternative to Moringa seed for water treatment and should be investigated further.
The interplay between atmospheric corrosion and antimicrobial efficiency of bare Cu and Cu5Zn5Al1Sn was studied upon exposures simulating high-touch surface conditions. The survival of the bacteria Bacillus subtilis during surface contact with Cu and Cu5Zn5Al1Sn was examined under different degrees of surface oxidation, tarnishing, wettability and copper ion release. Depending on surface conditions complete bacteria inhibition was obtained within 4 min on Cu and within 6-10 min on Cu5Zn5Al1Sn. The antibacterial efficiency increases slightly with copper release rate and is governed by complex interactions between the corroded metal surface, bacteria and extracellular polymeric substances produced by the bacteria.
Metal-based high-touch surfaces used for indoor applications such as doorknobs, light switches, handles and desks need to remain their antimicrobial properties even when tarnished or degraded. A novel laboratory methodology of relevance for indoor atmospheric conditions and fingerprint contact has therefore been elaborated for combined studies of both tarnishing/corrosion and antimicrobial properties of such high-touch surfaces. Cu metal was used as a benchmark material. The protocol includes pre-tarnishing/corrosion of the high touch surface for different time periods in a climatic chamber at repeated dry/wet conditions and artificial sweat deposition followed by the introduction of bacteria onto the surfaces via artificial sweat droplets. This methodology provides a more realistic and reproducible approach compared with other reported procedures to determine the antimicrobial efficiency of high-touch surfaces. It provides further a possibility to link the antimicrobial characteristics to physical and chemical properties such as surface composition, chemical reactivity, tarnishing/corrosion, surface roughness and surface wettability. The results elucidate that bacteria interactions as well as differences in extent of tarnishing can alter the physical properties (e.g. surface wettability, surface roughness) as well as the extent of metal release. The results clearly elucidate the importance to consider changes in chemical and physical properties of indoor hygiene surfaces when assessing their antimicrobial properties.
The name Arsenic is derived from the Greek word arsenikon, meaning potent. This element occurs in the ecosystem in different oxidation states of which As(III) and As(V) are most common to humans, animals, plant species. As is present in drinking water and soil from natural sources as well as a pollutant from agricultural and industrial processes. Differences in arsenic uptake by different plant species is controlled by many factors such as root surface area, root exudates, and rate of evapotranspiration. Some plant species have high affinity to accumulate arsenic in tissues above ground. Hyperaccumulator plants have a threshold arsenic content above 1,000 μg g-1 DM. We review bioremediation studies with especial emphasis on biosorption research on different arsenic species, plants and their biomass, agricultural and industry wastes, as well as the biomass of some fungi species. Bioremediation is considered as an alternative technique for the removal of As in groundwater. One of the popular methods among bioremediation techniques, phytoremediation uses living plants to remove arsenic from the environment or to render it less toxic, in bioaccumulation processes. Phytoremediation techniques often do not take into account the biosorption processes of living plants and plant litter. In biosorption techniques, contaminants can be removed by a biological substrate as a sorbent such as bacteria, fungi, algae or vascular plants. Bioremediation assures in situ treatment of polluted soils. Biosorption characteristics, equilibrium and kinetics of different biosorbents have also been addressed here. Evaluation of the current literature suggests that arsenic bioavailability and molecular level phytoremediation processes in bioremediation are crucial for designing phytoremediation technologies with improved, predictable remedial success.
Lake Pastos Grandes in Bolivia is mainly composed of salt flats, which are sporadically and only partially submerged during the wet season. In the present study, the chemical composition of water samples of the lake and some influent rivers was determined. We found that it is likely that the lake was influenced by the dilution of metals from ancient evaporites. We performed the first metagenomic studies of this lake. Analyses of shotgun metagenomics revealed that the relative abundances of Burkholderiales and Pseudomonadales were noteworthy in the water samples, whereas the archaea belonging to the Halobacteriales and Cyanobacteria from subsection III had high abundances in the salt flat. The eukaryotes Crustacea and Diatomea exhibited the highest abundances in the water samples. We investigated further the potential effect of human activities on the nitrogen cycle mobilization in the lake and the propagation of antimicrobial resistance genes. This is the first report about the cycle in the lake. Additionally, rifamycin resistance genes and genes related to efflux pumps, which are not considered a hazard when identified in metagenomes, had the uppermost relative abundances in all sampling points. We found that Lake Pastos Grandes hitherto does not show an appreciable influence by anthropogenic actions. The microbiome of Lake Pastos Grandes, including microbial distribution, the nitrogen cycle and antibiotic resistance genes, was analyzed.
The removal efficiency of diclofenac and ketoprofen for five fungal strains was examined. The synergistic effect between selected fungal strains using synthetic wastewater media was studied using both individual and mixed strains. The mixed cultures had no additional effect in reducing diclofenac and ketoprofen. In contrary, fungi removed pharmaceutical substances faster when growing individually. The results showed that T. versicolor monoculture removed diclofenac completely (>99.9 %) in non-sterile municipal wastewater within three hours of incubation and continued to show complete reduction within 14 days using fungal biofilm carriers. The mechanism of removal was proposed based on the laccase enzyme production and biosorption study which revealed that T. versicolor could used in both processes. Additionally, this study demonstrated that T. versicolor could be stable in the biofilm carriers, and thus able to compete with other microorganisms in wastewater to be a promising candidate to remove diclofenac from municipal wastewater during the treatment process.
Fungi have been shown to be good candidates to remove pharmaceuticals. However, the pilot-scale application mainly deals with the strict growth conditions and competition between microbial communities from wastewater. Thus, the bioaugmentation for removing the pharmaceuticals in municipal wastewater by Trametes versicolor and Aspergillus luchuensis using fluidized bed pelleted bioreactor was studied. To find the optimal performance of bioaugmentation by both fungi periodical biomass volume, 10 and 50 g per 1.25 L bioreactor, were tested. The removal of nutrients, pharmaceutical substances, and changes in the pH value, laccase activity, and total cell number of microorganisms were analyzed. The results showed that bioaugmentation has an effect in removing NH4-N and lower accumulation of NO3-N, whereas PO4−3-P and TOC did not show a significant effect on the reduction of nutrient load compared to the control without fungal addition. The results of removal efficiency for diclofenac, carbamazepine, and sulfamethoxazole showed that there was no significant effect on the removal while the highest removal efficiency (> 90 %) for ibuprofen, ketoprofen, and metoprolol was achieved by both fungi with periodical addition with 50 g of biomass after an incubation time of 3 hours. Finally, the data analysis with AI-based experimental design indicated that A. luchuensis can be a useful fungus for pharmaceutical removal and implies a perspective approach for optimization of fluidized bed pelleted bioreactor. Overall, the obtained results and the use of an AI-based platform is a promising approach for the optimization and operation of fluidized bed pelleted bioreactors.
Fungi have been shown to be effective candidates to remove pharmaceuticals. However, the pilot-scale application mainly deals with the strict growth conditions and competition between microbial community from wastewater. Thus, the bioaugmentation for removing the pharmaceuticals in municipal wastewater by T. versicolor and A. luchuensis using fluidized bed pelleted bioreactor was studied. To find the optimal performance of bioaugmentation two periodical biomass volume, 10 and 50 g per 1.25 L bioreactor, were tested. The removal of nutrients, pharmaceutical substances and changes in the pH value, laccase activity, and total microorganisms were analyzed. The results showed that bioaugmentation has an effect in removing NH 4 -N and lower accumulation of NO 3 -N, whereas PO 4 -3 -P and TOC did not show significant effect on the reduction of nutrient load. The results of removal efficiency for diclofenac, carbamazepine, and sulfamethoxazole showed that there was no significant effect on the removal while the highest removal efficiency (> 90%) for ibuprofen, ketoprofen, and metoprolol was achieved by both fungi with periodical addition with 50 g of biomass after an incubation time of 3 hours. Finally, the data analysis with AI-based experimental design indicated that A. luchuensis can be a promising fungus for pharmaceutical removal and implies as a promising approach for optimization of fluidized bed pelleted bioreactor. Overall, the obtained results and the use of an AI-based platform are a promising approach for optimization and operation of fluidized bed pelleted bioreactor.
Discharge of organic load from treated wastewater may cause environmental eutrophication. Recently, fungi have gained much attention due to their removal of pharmaceutical substances by enzymatic degradation and adsorption. However, the fungal effect in removing nutrients is less investigated. Therefore, two fungal species, the white-rot fungus T. versicolor as a laboratory strain and the mold A. luchuensis as an environmental isolate from the municipal wastewater treatment plant, were studied to determine the fungal potential for phosphorus, nitrogen, and the total organic carbon removal from municipal wastewater, carrying out a batch scale experiment to a fluidized bed pelleted bioreactor. During the batch scale experiment, the total removal (99.9 %) of phosphorus by T. versicolor was attained after a 6 hours-long incubation period while the maximal removal efficiency (99.9 %) for phosphorus from A. luchuensis was gained after an incubation period of 24 hours. Furthermore, both fungi showed that the pH adjustment to 5.5 kept the concentration of nitrogen constant and stabilized the total organic carbon reduction process for the entire incubation period. The results from the fluidized bed bioreactor demonstrated opposite tendencies on a nutrient removal comparing to a batch experiment where no significant effect on phosphorus, nitrogen, and total organics carbon reduction was observed. The obtained results from this study of batch and fluidized bed bioreactor experiments are a promising starting point for a successful fungal treatment optimization and application to wastewater treatment.
Though culture-independent methods have been used in preference to traditional isolation techniques for characterization of microbial community of wastewater treatment plants, it is difficult to widely apply this approach in resource-poor countries. The present study aimed to develop a test to identify the culturable portion of bacterial community in a high-strength wastewater. Wastewater samples were collected from nitrification-denitrification and settling tanks of the treatment plant of Elmo Leather AB tannery located in Boras, Sweden. After cultivating on nutrient agar with the optimal dilution (10(-2)), phenotypic and biochemical identification of the bacteria were done with colony morphology, Gram reaction, growth on MacConkey, phenylethanol media, triple sugar Iron agar slants, catalase and oxidase tests. Biochemical grouping of the isolates was done based on their test results for MacConkey, phenylethanol media, triple sugar Iron agar and oxidase test reaction. From the biochemical groups, isolates were randomly selected for API test and 16SrRNA gene sequencing. The isolates from the denitrification, nitrification tank were identified to be Paracoccus denitrificans (67%), Azoarcus spp (3%) and Spingomonas wittichii (1%). From the settling tank, Paracoccus denitrificans (22%), Corynebacterium freneyi (20%) and Bacillus cereus (1%) were identified. The grouping based on biochemical test results as well as the identification based on sequencing has shown coherence except for discrepancies with the API test. The preliminary implications of the grouping based on culture-based characteristics and its potential application for resource-limited environmental microbial studies is discussed.
Antisense peptide nucleic acids (PNA) can inhibit bacterial gene expression with gene and sequence specificity. Using attached carrier peptides that aid cell permeation, the antisense effects when targeting essential genes are sufficient to prevent growth and even kill bacteria. However, many design uncertainties remain, including the difficult question of target sequence selection. In this study, we synthesized 90 antisense peptide-PNAs to target sequences in a head to tail manner across the entire length of the mRNA encoding beta-lactamase. The results from this scan pointed to the start codon region as most sensitive to inhibition. To confirm and refine the result, a higher-resolution scan was conducted over the start codon region of the beta-lactamase gene and the essential Escherichia coli acpP gene. For both genes, the start codon region, including the Shine-Dalgarno motif, was sensitive, whereas antisense agents targeted outside of this region were largely ineffective. These results are in accord with natural antisense mechanisms, which typically hinder the start codon region, and the sensitivity of this region should hold true for most bacterial genes as well as for other RNase H-independent antisense agents that rely on a steric blocking mechanism. Therefore, although other design parameters are also important, the start codon region in E. coli mRNA is the most reliable target site for antisense PNAs.
A better understanding of the oviposition behaviour of malaria vectors might facilitate the development of new vector control tools. However, the factors that guide the aquatic habitat selection of gravid females are poorly understood. The present study explored the relative attractiveness of similar artificial ponds (0.8 m(2)) aged at varying lengths prior to opening in such a way that wild Anopheles arabiensis could choose between ponds that were freshly set up, or were aged 4 or 17 days old, to lay eggs. Physicochemical parameters, bacterial profile and volatile organic compounds emitted from ponds were investigated over three experimental rounds. Fresh ponds contained on average twice as many An. arabiensis instar larvae (mean 50, 95% confidence interval (CI) = 29-85) as the ponds that had aged 4 days (mean = 24, 95% CI = 14-42) and 17 days (mean = 20, 95% CI: 12-34). Fresh ponds were associated with a significantly higher turbidity combined with higher water temperature, higher nitrite levels and a lower pH and chlorophyll level than the older ponds. Round by round analyses suggested that bacteria communities differed between age groups and also that 4-heptanone, 2-ethylhexanal and an isomer of octenal were exclusively detected from the fresh ponds. These characteristics may be useful with respect to developing attract and kill strategies for malaria vector control.
Background: Cedrol, a sesquiterpene alcohol, is the first identified oviposition attractant for African malaria vectors. Finding the natural source of this compound might help to elucidate why Anopheles gambiae and Anopheles arabiensis prefer to lay eggs in habitats containing it. Previous studies suggest that cedrol may be a fungal metabolite and the essential oil of grass rhizomes have been described to contain a high amount of different sesquiterpenes. Results: Rhizomes of the grass Cyperus rotundus were collected in a natural malaria mosquito breeding site. Two fungi were isolated from an aqueous infusion with these rhizomes. They were identified as Fusarium falciforme and a species in the Fusarium fujikuroi species complex. Volatile compounds were collected from the headspace above fungal cultures on Tenax traps which were analysed by gas chromatography-mass spectrometry (GCMS). Cedrol and a cedrol isomer were detected in the headspace above the F. fujikuroi culture, while only cedrol was detected above the F. falciforme culture. Conclusion: Cedrol an oviposition attractant for African malaria vectors is produced by two fungi species isolated from grass rhizomes collected from a natural mosquito breeding site.
A promising sustainable strategy to valorize cellulose to high-value adsorbents for trace pharmaceuticals, like diclofenac sodium (DCF), in the water is demonstrated. Carbon nanospheres (CN) as the DCF adsorbent were derived from cellulose through a one-pot microwave-assisted hydrothermal carbonization method. CN exhibited efficient DCF removal (100% removal of 0.001 mg/mL DCF in 30 s and 59% removal of 0.01 mg/mL DCF in 1 h). The adsorption kinetics and isotherm data were well-fitted with the pseudo-second-order kinetic model and Langmuir model, respectively. The adsorption process was endothermic and spontaneous as confirmed by the thermodynamic parameters. Multiple characterization techniques including SEM/EDS, FTIR, FTIR-imaging and zeta potential were applied to qualitatively investigate the adsorption process. π-π stacking and hydrogen bonding were proposed as the dominant adsorption interactions. CN also demonstrated effective adsorption capacity towards three other commonly-detected contaminants in the wastewater including ketoprofen (KP), benzophenone (BZP), and diphenylamine (DPA), each bearing partial structural similarity with DCF. The affinity of the contaminants towards CN followed the order DPA > BZP > DCF > KP, which could be explained by the different configurations and chemical units. It was speculated that for DCF and KP, the steric hindrance and electrostatic repulsion produced by dissociated carboxyl groups can impede the adsorption process as compared to DPA and BZP. This methodology could offer further insights into the drug adsorption on the cellulose-derived carbon adsorbents and the use of bioderived carbons for treatment of wastewaters contaminated with pharmaceuticals.
Stringent microbial cell barriers limit the application of many substances in research and therapeutics. Carrier peptides that penetrate or translocate across cell membranes may help overcome this problem. To assess peptide-mediated delivery into two yeast and three bacterial species, a range of cell penetrating and signal peptide sequences were fused to green fluorescent protein (GFP), expressed in Escherichia coli, partially purified and incubated with growing cells. Fluorescence microscopy indicated several peptides that mediated delivery. In particular, VLTNENPFSDP efficiently delivered GFP into Candida albicans and Staphylococcus aureus, while YKKSNNPFSD was most efficient for Bacillus subtilis and CFFKDEL for Escherichia coli. Carrier peptides may improve delivery of certain large molecular mass molecules into microorganisms for research and therapeutic applications.
Microorganisms possess stringent cell membranes which limit the cellular uptake of antimicrobials. One strategy to overcome these barriers is to attach drugs or research reagents to carrier peptides that enter cells by passive permeation or active uptake. Here the short endocytosis signal peptide NPFSD was found to efficiently deliver both FITC and GFP,into Saccharomyces cerevisiae and Candida albicans with uptake into the majority of cells in a population. The NPFSD signal is itself non-toxic, but when fused to the ricin A chain toxin (RTA) the peptide enhanced both cell uptake and toxicity against C albicans, which like other yeasts is resistant to naked RTA. Cell entry required at least 1 h incubation, temperatures above 4degreesC, and an energy source, and uptake was outcompeted with free peptide. Therefore, the NPFSD peptide can carry a range of compounds into yeasts and this delivery route holds promise to enhance the activity of antifungals.
The present work compares the use of three flocculants for sedimentation of sludge and sludge water content from sewage wastewater i.e. magnetic iron oxide nanoparticles (MION), ferrous sulfate (chemical) and Moringa crude extract (protein). Sludge water content, wet/dry weight, turbidity and color were performed for, time kinetics and large-scale experiment. A 30% reduction of the sludge water content was observed when the wastewater was treated with either protein or chemical coagulant. The separation of sludge from wastewater treated with MION was achieved in less than 5. min using an external magnet, resulted in 95% reduction of sludge water content. Furthermore, MION formed denser flocs and more than 80% reduction of microbial content was observed in large volume experiments. The results revealed that MION is efficient in rapid separation of sludge with very low water content, and thus could be a suitable alternative for sludge sedimentation and dewatering in wastewater treatment processes.
The existing water treatment process often uses chemicals, which is of high health and environmental concern. The present study focused on the efficiency of microemulsion prepared magnetic iron oxide nanoparticles (ME-MIONs) and protein-functionalized nanoparticles (MOCP. +. ME-MIONs) in water treatment. Their influence on mineral ions and microorganisms present in the surface water from lake Brunnsviken and örlången, Sweden were investigated. Ion analysis of water samples before and after treatment with nanoparticles was performed. Microbial content was analyzed by colony forming units (CFU/ml). The results impart that ME-MIONs could reduce the water turbidity even in low turbid water samples. Reduction of microbial content (98%) was observed at 37 °C and more than 90% reduction was seen at RT and 30 °C when compared to untreated samples from lake örlången. The investigated surface water treatment method with ME-MIONs was not significantly affecting the mineral ion composition, which implies their potential complement in the existing treatment process.
The present study investigates the effective removal of phosphate in sewage wastewater using magnetic iron oxide nanoparticles (MION). The microemulsion-prepared magnetic iron oxide nanoparticles (ME-MION) of around 7-10 nm was synthesized using water-in-oil microemulsion method. The interaction of ME-MION and phosphate was studied using In situ FT-IR technique. Batch experiments were carried out with wastewater to determine the conc. and time efficiency using ME-MION for removal of phosphate. The vibration peak at 1004 cm-1 and the presence of hydroxyl group (OH-) at 3673 cm-1 confirms the binding of phosphate to ME-MION. ME-MION with 0.44 g L-1 exhibited more than 95% phosphate reduction in 5 min and close to 100% in 20 min. Conversely the experimental data obtained has been fitted with Langmuir isotherm model and also exhibited high correlation coefficients. The ME-MION was regenerated and can be reused for minimum 5 consecutive times. Efficient and fast reduction of phosphate was attained while the recovery of nanoparticles was achieved by an external magnetic field. To the author's knowledge, this is the first report that underscores around 100% phosphate removal from wastewater using ME-MION in 20 min. The approach utilized in this study offers a potential technique in the reduction of phosphate in wastewater whilst, reducing the time and reuse of nanoparticles.
Nanotechnology is considered as one of the key techniques that provide unique materials with high reactivity due to large surface to volume ratio and which could address the fundamental issues in water sector and environment. The present study investigates the efficiency of magnetic iron oxide nanoparticles in wastewater treatment based on Central Composite Face centered (CCF) matrix of response surface methodology for the reduction of turbidity and total nitrogen. The multiple linear regression fit (MLR) obtained for turbidity (r2 0.97) and total nitrogen reduction (r 0.94) supports the future predictions obtaining a significant model. The maximum reduction of turbidity and total nitrogen achieved was 93% and 41% respectively. Other contaminants such as color, total organic carbon, nitrate and microbial content could be reduced. The present study reveals that magnetic property, time and reduction of pollutants by magnetic nanoparticles could impart an efficient treatment process.
The increased levels of organic carbon in sewage wastewater during recent years impose a great challenge to the existing wastewater treatment process (WWTP). Technological innovations are therefore sought that can reduce the release of organic carbon into lakes and seas. In the present study, magnetic nanoparticles (NPs) were synthesized, functionalized with poly(ethylenimine) (PEI), and characterized using TEM (transmission electron microscopy), X-ray diffraction (XRD), FTIR (Fourier transform infrared spectroscopy), CCS (confocal correlation spectroscopy), SICS (scattering interference correlation spectroscopy), magnetism studies, and thermogravimetric analysis (TGA). The removal of total organic carbon (TOC) and other contaminants using PEI-coated magnetic nanoparticles (PEI-NPs) was tested in wastewater obtained from the Hammarby Sjöstadsverk sewage plant, Sweden. The synthesized NPs were about 12 nm in diameter and showed a homogeneous particle size distribution in dispersion by TEM and CCS analyses, respectively. The magnetization curve reveals superparamagnetic behavior, and the NPs do not reach saturation because of surface anisotropy effects. A 50% reduction in TOC was obtained in 60 min when using 20 mg/L PEI-NPs in 0.5 L of wastewater. Along with TOC, other contaminants such as turbidity (89%), color (86%), total nitrogen (24%), and microbial content (90%) were also removed without significant changes in the mineral ion composition of wastewater. We conclude that the application of PEI-NPs has the potential to reduce the processing time, complexity, sludge production, and use of additional chemicals in the WWTP.