Effect of the Application of Ochrobactrum sp.-Immobilised Biochar on the Remediation of Diesel-Contaminated Soil.

The immobilisation of bacteria on biochar has shown potential for enhanced remediation of petroleum hydrocarbon-contaminated soil. However, there is a lack of knowledge regarding the effect of bacterial immobilisation on biosolids-derived biochar for the remediation of diesel-contaminated soil. This current study aimed to assess the impact of the immobilisation of an autochthonous hydrocarbonoclastic bacteria, Ochrobacterium sp. (BIB) on biosolids-derived biochar for the remediation of diesel-contaminated soil. Additionally, the effect of fertiliser application on the efficacy of the BIB treatment was investigated. Biochar (BC) application alone led to significantly higher hydrocarbon removal than the control treatment at all sampling times (4887-11,589 mg/kg higher). When Ochrobacterium sp. was immobilised on biochar (BIB), the hydrocarbon removal was greater than BC by 5533 mg/kg and 1607 mg/kg at weeks 10 and 22, respectively. However, when BIB was co-applied with fertiliser (BIBF), hydrocarbon removal was lower than BIB alone by 6987-11,767 mg/kg. Quantitative PCR (q-PCR) analysis revealed that the gene related to Ochrobacterium sp. was higher in BIB than in the BC treatment, which likely contributed to higher hydrocarbon removal in the BIB treatment. The results of the q-PCR analysis for the presence of alkB genes and FTIR analysis suggest that the degradation of alkane contributed to hydrocarbon removal. The findings of this study demonstrate that bacterial immobilisation on biosolids-derived biochar is a promising technique for the remediation of diesel-contaminated soil. Future studies should focus on optimising the immobilisation process for enhanced hydrocarbon removal.

Dead in the water - Role of relic DNA and primer choice for targeted sequencing surveys of anaerobic sewage sludge intended for biological monitoring.

DNA-based monitoring of microbial communities that are responsible for the performance of anaerobic digestion of sewage wastes has the potential to improve resource recoveries for wastewater treatment facilities. By treating sludge with propidium monoazide (PMA) prior to amplicon sequencing, this study explored how the presence of DNA from dead microbial biomass carried over with feed sludge may mislead process-relevant biomarkers, and whether primer choice impacts such assessments. Four common primers were selected for amplicon preparation, also to determine if universal primers have sufficient taxonomic or functional coverage for monitoring ecological performance; or whether two domain-specific primers for Bacteria and Archaea are necessary. Anaerobic sludges of three municipal continuously stirred-tank reactors in Victoria, Australia, were sampled at one time-point. A total of 240 amplicon libraries were sequenced on a Miseq using two universal and two domain-specific primer pairs. Untargeted metabolomics was chosen to complement biological interpretation of amplicon gene-based functional predictions. Diversity, taxonomy, phylogeny and functional potentials were systematically assessed using PICRUSt2, which can predict community wide pathway abundance. The two chosen universal primers provided similar diversity profiles of abundant Bacteria and Archaea, compared to the domain-specific primers. About 16 % of all detected prokaryotic genera covering 30 % of total abundances and 6 % of PICRUSt2-estimated pathway abundances were affected by PMA. This showed that dead biomass in the anaerobic digesters impacted DNA-based assessments, with implications for predicting active processes, such as methanogenesis, denitrification or the identification of organisms associated with biological foams. Hence, instead of running two sequencing runs with two different domain-specific primers, we propose conducting PMA-seq with universal primer pairs for routine performance monitoring. However, dead sludge biomass may have some predictive value. In principal component analysis the compositional variation of 239 sludge metabolites resembled that of 'dead-plus-alive' biomass, suggesting that dead organisms contributed to the potentially process-relevant sludge metabolome.

Assessing the impact of immobilisation on the bioavailability of PFAS to plants in contaminated Australian soils.

Per- and polyfluoroalkyl substances (PFAS) have become a key concern to both environmental and human health due to their extreme persistence in the environment and their ability to bioaccumulate in plants, animals, and humans. In this mesocosm study, Australian PFAS-contaminated soil with a mean total concentration of 8.05 mg/kg and a mean combined PFHxS + PFOS concentration of 7.89 mg/kg was treated with an immobilisation sorbent (RemBind®) at different application rates (0.5, 1, 1.5, 2, 3, 4, and 5% w/w). To assess the efficacy of this immobilisation treatment, PFAS leachability, PFAS plant uptake, and ecotoxicity tests were conducted. Leachability testing was performed according to the Australian Standard Leaching Procedure (ASLP) at pH 5 and 7. A grass species (Dactylis glomerata) was used to measure plant uptake of PFAS from untreated and treated contaminated soil. In addition, the Microtox test was used to assess the associated ecotoxicity. The immobilisation treatment resulted in a significant reduction of 88.5-99.8% in the total PFAS leachability and 88.7-99.8% in the combined PFOS and PFHxS leachability at pH 5. Similarly, significant reductions (5-12-fold) were observed in the plant uptake of total PFAS and combined PFOS and PFHxS in all treated soil samples. In addition, although the Microtox test showed relatively low ecotoxicity in all the experimental samples, including the untreated soil, a significant decrease in the ecotoxicity of treated soil samples was observed. The results from this study highlight that this treatment approach has the potential to reduce both PFAS leachability and plant bioavailability with a relatively low associated ecotoxicity. This is likely to reduce the risk of the transfer of PFAS into higher trophic levels. This immobilisation treatment may, therefore, reduce the risk associated with PFAS-contaminated soils and may be an important remediation tool for managing certain PFAS-contaminated soils.

Wood biochar enhances methanogenesis in the anaerobic digestion of chicken manure under ammonia inhibition conditions.

The process of breaking down chicken manure through anaerobic digestion is an effective waste management technology. However, chicken manure can be a challenging feedstock, causing ammonia stress and digester instability. This study examined the impacts of adding wood biochar and acid-alkali-treated wood biochar to anaerobically digest chicken manure under conditions of ammonia inhibition. The results highlighted that only the addition of 5 % acid-alkali-treated wood biochar by volume can achieve cumulative methane production close to the typical methane potential range of chicken manure. The treated wood biochar also exhibited highest total ammonia nitrogen removal compared to the Control treatment. Scanning Electron Microscope revealed growing interactions between biochar and methanogens over time. Real-time polymerase chain reaction showed that treated wood biochar produced the highest number of bacterial biomass. In addition, 16S amplicon-based sequencing identified a more robust archaeal community from treated biochar addition. Overall, the acid-alkali treatment of biochar represents an effective method of modifying biochar to improve its performance in anaerobic digestion.

Prospective Subunit Nanovaccine against Mycobacterium tuberculosis Infection─Cubosome Lipid Nanocarriers of Cord Factor, Trehalose 6,6' Dimycolate.

An improved vaccine is urgently needed to replace the now more than 100-year-old Bacillus Calmette-Guérin (BCG) vaccine against tuberculosis (TB) disease, which represents a significant burden on global public health. Mycolic acid, or cord factor trehalose 6,6' dimycolate (TDM), a lipid component abundant in the cell wall of the pathogen Mycobacterium tuberculosis (MTB), has been shown to have strong immunostimulatory activity but remains underexplored due to its high toxicity and poor solubility. Herein, we employed a novel strategy to encapsulate TDM within a cubosome lipid nanocarrier as a potential subunit nanovaccine candidate against TB. This strategy not only increased the solubility and reduced the toxicity of TDM but also elicited a protective immune response to control MTB growth in macrophages. Both pre-treatment and concurrent treatment of the TDM encapsulated in lipid monoolein (MO) cubosomes (MO-TDM) (1 mol %) induced a strong proinflammatory cytokine response in MTB-infected macrophages, due to epigenetic changes at the promoters of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) in comparison to the untreated control. Furthermore, treatment with MO-TDM (1 mol %) cubosomes significantly improved antigen processing and presentation capabilities of MTB-infected macrophages to CD4 T cells. The ability of MO-TDM (1 mol %) cubosomes to induce a robust innate and adaptive response in vitro was further supported by a mathematical modeling study predicting the vaccine efficacy in vivo. Overall, these results indicate a strong immunostimulatory effect of TDM when delivered through the lipid nanocarrier, suggesting its potential as a novel TB vaccine.

Cubosome Lipid Nanocarriers As a Drug Delivery Vehicle for Intracellular Mycobacterium tuberculosis Infections.

Mycobacterium tuberculosis (MTB) causes the infectious disease tuberculosis (TB), responsible for more deaths than any other single infectious disease in history. Intracellular MTB are slow growing and difficult to target with traditional antitubercular drugs, leading to the emergence of multidrug resistance in TB infection, which is a major global public health issue. Recent advances in innovative lipid nanotechnologies for drug delivery have demonstrated promising outcomes for chronic infectious diseases but have not yet been tested as potential delivery systems for intracellular infections such as TB. The current study evaluates the potential of monoolein (MO)-based cationic cubosomes for the encapsulation and delivery of the first line antitubercular drug rifampicin (RIF) against an MTB-H37Ra in vitro culture model. In particular, we show that the use of cationic cubosomes as delivery vehicles reduced the minimum inhibitory concentration (MIC) of RIF by 2-fold against actively replicating MTB-H37Ra (compared to that of the free drug) and also shortened the lifecycle duration of axenic MTB-H37Ra from 5 to 3 days. The cubosome-mediated delivery was also found to be effective against intracellular MTB-H37Ra within THP-1 human macrophages, with a 2.8 log reduction in viability of the bacilli after 6 days incubation at the MIC. The killing time was also reduced from 8 to 6 days without distressing the host macrophages. Mechanistic studies on the uptake of RIF-loaded cationic cubosomes using total internal reflection fluorescence microscopy (TIRFM) demonstrated the capacity of these lipid particles to effectively target intracellular bacteria. Overall, these results demonstrate that cationic cubosomes are a potent delivery system for the antitubercular drug RIF for therapeutic management of TB.

Development of A Rapid, Low-Cost Portable Detection Assay for Enterococci in Wastewater and Environmental Waters.

Waterborne diseases are known as a leading cause of illness and death in both developing and developed countries. Several pathogens can be present in contaminated water, particularly waters containing faecal material; however, routine monitoring of all pathogens is not currently possible. Enterococcus faecalis, which is present in the microflora of human and animals has been used as a faecal indicator in water due to its abundance in surface water and soil. Accurate and fast detection methods are critical for the effective monitoring of E. faecalis in the environment. Although conventional and current molecular detection techniques provide sufficient sensitivity, specificity and throughput, their use is hampered by the long waiting period (1-6 days) to obtain results, the need for expensive laboratory equipment, skilled personnel, and cold-chain storage. Therefore, this study aimed to develop a detection system for E. faecalis that would be simple, rapid, and low-cost, using an isothermal DNA amplification assay called recombinase polymerase amplification (RPA), integrated with a lateral flow assay (LFA). The assay was found to be 100% selective for E. faecalis and capable of detecting rates as low as 2.8 × 103 cells per 100 mL from water and wastewater, and 2.8 × 104 cells per 100 mL from saline water. The assay was completed in approximately 30 min using one constant temperature (38 °C). In addition, this study demonstrated the quantitation of E. faecalis using a lateral flow strip reader for the first time, enhancing the potential use of RPA assay for the enumeration of E. faecalis in wastewater and heavily contaminated environmental waters, surface water, and wastewater. However, the sensitivity of the RPA-LFA assay for the detection of E. faecalis in tap water, saline water and in wastewater was 10-1000 times lower than that of the Enterolert-E test, depending on the water quality. Nevertheless, with further improvements, this low-cost RPA-LFA may be suitable to be used at the point-of-need (PON) if conjugated with a rapid field-deployable DNA extraction method.

Investigations into valorisation of trade wastewater for biomethane production.

Biogas production from wastewater is one way that industrial sites can work towards the UN Sustainable Development Goals, while recovering a valuable resource. The objective of this study was to investigate the suitability of data collected by municipal wastewater service providers as a method of classifying and screening waste producers as potential sites for biogas resource recovery by anaerobic digestion. Industrial wastewater samples, including raw effluent and treated waste ready for discharge, were examined, and biomethane potential assays performed. Results of chemical analysis and lab-scale digestion were compared to historical service provider data, and patterns were observed. Biomethane yields of up to 357 mL/gVS and 287mL/gVS were achieved from raw and treated effluent respectively. Digestion at the top four prospects could produce over 4690 GJ of methane and save $47,000 in natural gas costs, offsetting 490 tonnes of CO2 equivalent annually. These streams, from logistics, waste management, food and animal product businesses, combined high levels of degradable substrates and low levels of inhibitory components. While it is unlikely that this type of screening program can be completely accurate, certain parameters, including high sodium concentration, are applicable for discounting the potential for biogas production. This knowledge can be a valuable tool in the process of selecting sites for future resource recovery, therefore increasing the uptake of these processes, resulting in economic, environmental, and climate change mitigation benefits.

The co-application of biochar with bioremediation for the removal of petroleum hydrocarbons from contaminated soil.

Soil pollution from petroleum hydrocarbon is a global environmental problem that could contribute to the non-actualisation of the United Nations Sustainable Development Goals. Several techniques have been used to remediate petroleum hydrocarbon-contaminated soils; however, there are technical and economical limitations to existing methods. As such, the development of new approaches and the improvement of existing techniques are imperative. Biochar, a low-cost carbonaceous product of the thermal decomposition of waste biomass has gained relevance in soil remediation. Biochar has been applied to remediate hydrocarbon-contaminated soils, with positive and negative results reported. Consequently, attempts have been made to improve the performance of biochar in the hydrocarbon-based remediation process through the co-application of biochar with other bioremediation techniques as well as modifying biochar properties before use. Despite the progress made in this domain, there is a lack of a detailed single review consolidating the critical findings, new developments, and challenges in biochar-based remediation of petroleum hydrocarbon-contaminated soil. This review assessed the potential of biochar co-application with other well-known bioremediation techniques such as bioaugmentation, phytoremediation, and biostimulation. Additionally, the benefits of modification in enhancing biochar suitability for bioremediation were examined. It was concluded that biochar co-application generally resulted in higher hydrocarbon removal than sole biochar treatment, with up to a 4-fold higher removal observed in some cases. However, most of the biochar co-applied treatments did not result in hydrocarbon removal that was greater than the additive effects of individual treatment. Overall, compared to their complementary treatments, biochar co-application with bioaugmentation was more beneficial in hydrocarbon removal than biochar co-application with either phytoremediation or biostimulation. Future studies should integrate the ecotoxicological and cost implications of biochar co-application for a viable remediation process. Lastly, improving the synergistic interactions of co-treatment on hydrocarbon removal is critical to capturing the full potential of biochar-based remediation.

Remediation of groundwater contaminated with dye using carbon dots technology: Ecotoxicological and microbial community responses.

Groundwater pollution poses a serious threat to the main source of clean water globally. Nanoparticles have the potential for remediation of polluted aquifers; however, environmental safety concerns associated with in situ deployments of such technology include potential detrimental effects on microorganisms in terms of toxicity and functional disruptions. In this work, we evaluated a new and ecofriendly approach using carbon dots (CDs) as Fenton-like catalysts to catalyse the degradation of dye-containing groundwater samples. This investigation aimed at evaluating the efficacy of a novel remediation technology in terms of dye degradation and toxicity reduction while assessing its impacts on aquatic microorganisms. Uncontaminated Australian groundwater samples were spiked with methylene blue and incubated in the dark, at 18 °C, under slow agitation, using CDs at 0.5 mg mL-1 and H2O2 at 73.5 mM for 25 h. The dye degradation rate was determined as well as the toxicity of the treated solutions using the Microtox® bioassay. Further, to determine the changes in the groundwater microbial community, 16 S rRNA sequencing was used and evenness and diversity indices were analysed using Pielou's evenness and Simpson index, respectively. This study revealed that dye-containing groundwater were effectively treated by CDs showing a degradation rate of 78-82% and a significant 4-fold reduction in the toxicity. Characterisation of the groundwater microbiota revealed a predominance of at least 60% Proteobacteria phylum in all samples where diversity and evenness were maintained throughout the remediation process. The results showed that CDs could be an efficient approach to treat polluted groundwater and potentially have minimum impact on the environmental microbiome.

Carbon Dot Therapeutic Platforms: Administration, Distribution, Metabolism, Excretion, Toxicity, and Therapeutic Potential.

Ultrasmall nanoparticles are often grouped under the broad umbrella term of "nanoparticles" when reported in the literature. However, for biomedical applications, their small sizes give them intimate interactions with biological species and endow them with unique functional physiochemical properties. Carbon quantum dots (CQDs) are an emerging class of ultrasmall nanoparticles which have demonstrated considerable biocompatibility and have been employed as potent theragnostic platforms. These particles find application for increasing drug solubility and targeting, along with facilitating the passage of drugs across impermeable membranes (i.e., blood brain barrier). Further functionality can be triggered by various environmental conditions or external stimuli (i.e., pH, temperature, near Infrared (NIR) light, ultrasound), and their intrinsic fluorescence is valuable for diagnostic applications. The focus of this review is to shed light on the therapeutic potential of CQDs and identify how they travel through the body, reach their site of action, administer therapeutic effect, and are excreted. Investigation into their toxicity and compatibility with larger nanoparticle carriers is also examined. The future of CQDs for theragnostic applications is promising due to their multifunctional attributes and documented biocompatibility. As nanomaterial platforms become more commonplace in clinical treatments, the commercialization of CQD therapeutics is anticipated.

Biodegradation of novel bioplastics made of starch, polyhydroxyurethanes and cellulose nanocrystals in soil environment.

Plastic pollution is recognized as a major environmental problem in many countries. Over the last decade, academics have embraced research on bioplastics to discover newer high-end green materials. However, the end-of-life environmental fate of such materials is not adequately understood. Non-isocyanate polyhydroxyurethanes (PHUs) are green engineering materials with huge potential to replace traditional polyurethanes. Despite this immense potential, a number of questions about their environmental fate remain unanswered. The present study investigated the extent and mechanisms underlying soil biodegradation of PHUs and determined whether the deterioration of PHUs within starch bioplastics (ST) can improve the biodegradation of starch (ST)-PHU hybrids. Soil microbiomes managed to effectively and quickly digest not only PHUs but also ST-PHU hybrids. All ST-PHU hybrids were characterized by exceptional biodegradability with mass losses of up to ~88% following a soil burial time of only 120 days. The biodegradation of ST-alone bioplastics was 69% under identical conditions. The presence of cellulose nanocrystals (CNC) reduced the potential for the soil microbial community to degrade nanohybrids (ST-PHU-CNC). Microbially digested bioplastics with PHU presented less stages of thermal degradation, and reduced intensities of FTIR, NMR and XPS signals compared to the original films, indicating improvement of the biodegradation mechanism. These findings suggested the positive environmental implications of PHU in improving the bioplastic's degradation and their potential for future applications.

Substrate-to-inoculum ratio drives solid-state anaerobic digestion of unamended grape marc and cheese whey.

Inoculation dose is a key operational parameter for the solid-state anaerobic digestion (SS-AD) of lignocellulosic biomass, maximum methane recovery, and stable digester performance. The novelty of this study was the co-digestion of unamended full-strength grape marc and cheese whey for peak methane extraction at variable inoculation levels. An acclimatised digestate from a preceding anaerobic treatment was used as a downstream inoculum. The impact of inoculum size (wet weight) was evaluated at 0/10, 5/5, 7/3 and 9/1 substrate-to-inoculum (S/I) ratios, corresponding to an initial concentration of 20-30% total solids (TS) in digesters over 58 days at 45°C. The optimal 7/3 S/I produced the highest cumulative methane yield, 6.45 L CH4 kg-1 VS, coinciding with the lowest initial salinity at 11%; the highest volumetric methane productivity rate of 0.289±0.044 L CH4 LWork-1 d-1; the highest average COD/N ratio of 9.88; the highest final pH of 9.13, and a maximum 15.07% elemental carbon removal; for a lag time of 9.4 days. This study identified an optimal inoculation dose and opens up an avenue for the direct co-digestion of grape marc and cheese whey without requirements for substrate pretreatment, thus improving the overall bioenergy profile of the winery and dairy joint resource recovery operations.

The role of microbial ecology in improving the performance of anaerobic digestion of sewage sludge.

The use of next-generation diagnostic tools to optimise the anaerobic digestion of municipal sewage sludge has the potential to increase renewable natural gas recovery, improve the reuse of biosolid fertilisers and help operators expand circular economies globally. This review aims to provide perspectives on the role of microbial ecology in improving digester performance in wastewater treatment plants, highlighting that a systems biology approach is fundamental for monitoring mesophilic anaerobic sewage sludge in continuously stirred reactor tanks. We further highlight the potential applications arising from investigations into sludge ecology. The principal limitation for improvements in methane recoveries or in process stability of anaerobic digestion, especially after pre-treatment or during co-digestion, are ecological knowledge gaps related to the front-end metabolism (hydrolysis and fermentation). Operational problems such as stable biological foaming are a key problem, for which ecological markers are a suitable approach. However, no biomarkers exist yet to assist in monitoring and management of clade-specific foaming potentials along with other risks, such as pollutants and pathogens. Fundamental ecological principles apply to anaerobic digestion, which presents opportunities to predict and manipulate reactor functions. The path ahead for mapping ecological markers on process endpoints and risk factors of anaerobic digestion will involve numerical ecology, an expanding field that employs metrics derived from alpha, beta, phylogenetic, taxonomic, and functional diversity, as well as from phenotypes or life strategies derived from genetic potentials. In contrast to addressing operational issues (as noted above), which are effectively addressed by whole population or individual biomarkers, broad improvement and optimisation of function will require enhancement of hydrolysis and acidogenic processes. This will require a discovery-based approach, which will involve integrative research involving the proteome and metabolome. This will utilise, but overcome current limitations of DNA-centric approaches, and likely have broad application outside the specific field of anaerobic digestion.

Dynamic Effect of Operational Regulation on the Mesophilic BioMethanation of Grape Marc.

Wine production annually generates an estimated 11 million metric tonnes of grape marc (GM) worldwide. The diversion of this organic waste away from landfill and towards its use in the generation of renewable energy has been investigated. This study aimed to evaluate the effectiveness of operational parameters relating to the treatment regime and inoculum source in the extraction of methane from GM under unmixed anaerobic conditions at 35 °C. The study entailed the recirculation of a previously acclimated sludge (120 days) as downstream inoculum, an increased loading volume (1.3 kg) and a low substrate-to-inoculum ratio (10:3 SIR). The results showed that an incorporation of accessible operational controls can effectively enhance cumulative methane yield (0.145 m3 CH4 kg-1 VS), corresponding to higher amounts of digestible organics converted. The calculated average volumetric methane productivity equalled 0.8802 L CH4 LWork-1 d-1 over 33.6 days whilst moderate pollutant removal (43.50% COD removal efficiency) was achieved. Molecular analyses identified Firmicutes and Bacteroidetes phyla as core organisms for hydrolytic and fermentative stages in trophic relationships with terminal electron acceptors from the methane-producing Methanosarcina genus. Economic projections established that the cost-effective operational enhancements were sustainable for valorisation from grape marc by existing wineries and distilleries.

Natural attenuation of legacy hydrocarbon spills in pristine soils is feasible despite difficult environmental conditions in the monsoon tropics.

The Kimberley region of Western Australia is a National Heritage listed region that is internationally recognised for its environmental and cultural significance. However, petroleum spills have been reported at a number of sites across the region, representing an environmental concern. The region is also characterised as having low soil nutrients, high temperatures and monsoonal rain - all of which may limit the potential for natural biodegradation of petroleum. Therefore, this work evaluated the effect of legacy petroleum hydrocarbons on the indigenous soil microbial community (across the domains Archaea, Bacteria and Fungi) at three sites in the Kimberley region. At each site, soil cores were removed from contaminated and control areas and analysed for total petroleum hydrocarbons, soil nutrients, pH and microbial community profiling (using16S rRNA and ITS sequencing on the Illumina MiSeq Platform). The presence of petroleum hydrocarbons decreased microbial diversity across all kingdoms, altered the structure of microbial communities and increased the abundance of putative hydrocarbon degraders (e.g. Mycobacterium, Acremonium, Penicillium, Bjerkandera and Candida). Microbial community shifts from contaminated soils were also associated with an increase in soil nutrients (notably Colwell P and S). Our study highlights the long-term effect of legacy hydrocarbon spills on soil microbial communities and their diversity in remote, infertile monsoonal soils, but also highlights the potential for natural attenuation to occur in these environments.

The Impacts of Different Biological Treatments on the Transformation of Explosives Waste Contaminated Sludge.

The dinitrotoluene isomers 2,4 and 2,6-dinitrotoluene (DNT) represent highly toxic, mutagenic, and carcinogenic compounds used in explosive manufacturing and in commercial production of polyurethane foam. Bioremediation, the use of microbes to degrade residual DNT in industry wastewaters, represents a promising, low cost and environmentally friendly alternative technology to landfilling. In the present study, the effect of different bioremediation strategies on the degradation of DNT in a microcosm-based study was evaluated. Biostimulation of the indigenous microbial community with sulphur phosphate (2.3 g/kg sludge) enhanced DNT transformation (82% transformation, from 300 g/L at Day 0 to 55 g/L in week 6) compared to natural attenuation over the same period at 25 °C. The indigenous microbial activity was found to be capable of transforming the contaminant, with around 70% transformation of DNT occurring over the microcosm study. 16S rDNA sequence analysis revealed that while the original bacterial community was dominated by Gammaproteobacteria (30%), the addition of sulphur phosphate significantly increased the abundance of Betaproteobacteria by the end of the biostimulation treatment, with the bacterial community dominated by Burkholderia (46%) followed by Rhodanobacter, Acidovorax and Pseudomonas. In summary, the results suggest biostimulation as a treatment choice for the remediation of dinitrotoluenes and explosives waste.

Fluorescent Magnesium Hydroxide Nanosheet Bandages with Tailored Properties for Biocompatible Antimicrobial Wound Dressings and pH Monitoring.

Magnesium hydroxide (Mg(OH)2) is hailed as a cheap and biocompatible material with antimicrobial potential; however, research aimed at instilling additional properties and functionality to this material is scarce. In this work, we synthesized novel, fluorescent magnesium hydroxide nanosheets (Mg(OH)2-NS) with a morphology that closely resembles that of graphene oxide. These multifunctional nanosheets were employed as a potent antimicrobial agent against several medically relevant bacterial and fungal species, particularly on solid surfaces. Their strong fluorescence signature correlates to their hydroxide makeup and can therefore be used to assess their degradation and functional antimicrobial capacity. Furthermore, their pH-responsive change in fluorescence can potentially act as a pH probe for wound acidification, which is characteristic of healthy wound healing. These fluorescent antimicrobial nanosheets were stably integrated into biocompatible electrospun fibers and agarose gels to add functionality to the material. This reinforces the suitability of the material to be used as antimicrobial bandages and gels. The biocompatibility of the Mg(OH)2-NS for topical medical applications was supported by its noncytotoxic action on human keratinocyte (HaCaT) cells.

Application of Co-Culture Technology to Enhance Protease Production by Two Halophilic Bacteria, Marinirhabdus sp. and Marinobacter hydrocarbonoclasticus.

Although axenic microbial cultures form the basis of many large successful industrial biotechnologies, the production of single commercial microbial strains for use in large environmental biotechnologies such as wastewater treatment has proved less successful. This study aimed to evaluate the potential of the co-culture of two halophilic bacteria, Marinirhabdus sp. and Marinobacter hydrocarbonoclasticus for enhanced protease activity. The co-culture was significantly more productive than monoculture (1.6-2.0 times more growth), with Marinobacter hydrocarbonoclasticus being predominant (64%). In terms of protease activity, enhanced total activity (1.8-2.4 times) was observed in the co-culture. Importantly, protease activity in the co-culture was found to remain active over a much broader range of environmental conditions (temperature 25 °C to 60 °C, pH 4-12, and 10-30% salinity, respectively). This study confirms that the co-culturing of halophilic bacteria represents an economical approach as it resulted in both increased biomass and protease production, the latter which showed activity over arange of environmental conditions.

Review: Trends in point-of-care diagnosis for Escherichia coli O157:H7 in food and water.

Escherichia coli O157:H7, a Shiga-producing E. coli is a major pathogenic E. coli strain which since the early 1980s has become a crucial food and water-borne pathogen. Several management strategies can be applied to control the spread of infection; however early diagnosis represents the optimum preventive strategy to minimize the infection. Therefore, it is crucial to detect this pathogen in a fast and efficient manner in order to reduce the morbidity and mortality. Currently used gold standard tests rely on culture and pre-enrichment of E. coli O157:H7 from the contaminated source; they are time consuming and laborious. Molecular methods such as polymerase chain reaction are sensitive; however, they require expensive instrumentation. Therefore, there is a requirement for Accurate, Sensitive, Specific, User friendly, Rapid, Equipment free and Deliverable (ASSURED) detection methods for use in the laboratory and in the field. Emerging technologies such as isothermal amplification methods, biosensors, surface enhanced Raman Spectroscopy, paper-based diagnostics and smartphone-based digital methods are recognized as new approaches in the field of E. coli O157:H7 diagnostics and are discussed in this review. Mobile PCR and CRISPR-Cas diagnostic platforms have been identified as new tools in E. coli O157:H7 POC diagnostics with the potential for implementation by industry. This review describes advances and progress in the field of E. coli O157:H7 diagnosis in the context of food and water industry. The focus is on emerging high throughput point-of-care (POC) E. coli O157:H7 diagnostics and the requirement for the transformation to service routine diagnostics in the food and water industry.