Assessing sustainability of microalgae-based wastewater treatment: Environmental considerations and impacts on human health.

An integrated life cycle assessment (LCA) and quantitative microbial risk assessment (QMRA) were conducted to assess microalgae-mediated wastewater disinfection (M-WWD). M-WWD was achieved by replacing ultraviolet disinfection with a microalgal open raceway pond in an existing sewage treatment plant (STP) in India. Regarding impacts on human health, both M-WWD and STP yielded comparable life cycle impacts, around 0.01 disability-adjusted life years (DALYs) per person per year. However, QMRA impacts for M-WWD (0.053 DALYs per person per year) were slightly lower than that for STP while considering exposure to E. coli O157:H7 and adenovirus. Additionally, a comparative LCA resolved the dilemma about the appropriate utilization of microalgal biomass. Among biodiesel, biocrude, and biogas production, the lowest impacts of 0.015 DALYs per person per year were obtained for biocrude for 1 m3 water treated by M-WWD. Electricity consumption in microalgae cultivation was a major environmental hotspot. Overall, M-WWD, followed by production of microalgal biocrude, emerged as a sustainable alternative from environmental and public health perspectives. These findings set the foundation for pilot-scale M-WWD system development, testing, and economic evaluation. Such comprehensive investigations, encompassing LCA, QMRA, and resource recovery scenarios, offer crucial insights for stakeholders and decision-makers in wastewater treatment and environmental management.

Integrating life cycle assessment with quantitative microbial risk assessment for a holistic evaluation of sewage treatment plant.

An integrated approach was employed in the present study to combine life cycle assessment (LCA) with quantitative microbial risk assessment (QMRA) to assess an existing sewage treatment plant (STP) at Roorkee, India. The midpoint LCA modeling revealed that high electricity consumption (≈ 576 kWh.day-1) contributed to the maximum environmental burdens. The LCA endpoint result of 0.01 disability-adjusted life years per person per year (DALYs pppy) was obtained in terms of the impacts on human health. Further, a QMRA model was developed based on representative sewage pathogens, including E. coli O157:H7, Giardia sp., adenovirus, norovirus, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The public health risk associated with intake of pathogen-laden aerosols during treated water reuse in sprinkler irrigation was determined. A cumulative health risk of 0.07 DALYs pppy was obtained, where QMRA risks contributed 86 % of the total health impacts. The annual probability of illness per person was highest for adenovirus and norovirus, followed by SARS-CoV-2, E. coli O157:H7 and Giardia sp. Overall, the study provides a methodological framework for an integrated LCA-QMRA assessment which can be applied across any treatment process to identify the hotspots contributing maximum environmental burdens and microbial health risks. Furthermore, the integrated LCA-QMRA approach could support stakeholders in the water industry to select the most suitable wastewater treatment system and establish regulations regarding the safe reuse of treated water.

Genetic regulatory element based whole-cell biosensors for the detection of metabolic disorders.

Clinicians require simple, and cost-effective diagnostic tools for the quantitative determination of amino acids in physiological fluids for the detection of metabolic disorder diseases. Besides, amino acids also act as biological markers for different types of cancers and cardiovascular diseases. Herein, we applied an in-silico based approach to identify potential amino acid-responsive genetic regulatory elements for the detection of metabolic disorders in humans. Identified sequences were further transcriptionally fused with GFP, thus generating an optical readout in response to their cognate targets. Screening of genetic regulatory elements led us to discover two promoter elements (pmetE::GFP and ptrpL::GFP) that showed a significant change in the fluorescence response to homocysteine and tryptophan, respectively. The developed biosensors respond specifically and sensitively with a limit of detection of 3.8 µM and 3 µM for homocysteine and tryptophan, respectively. Furthermore, the clinical utility of this assay was demonstrated by employing it to identify homocystinuria and tryptophanuria diseases through the quantification of homocysteine and tryptophan in plasma and urine samples within 5 h. The precision and accuracy of the biosensors for disease diagnosis were well within an acceptable range. The general strategy used in this system can be expanded to screen different genetic regulatory elements present in other gram-negative and gram-positive bacteria for the detection of metabolic disorders.

Homogentisic Acid-Based Whole-Cell Biosensor for Detection of Alkaptonuria Disease.

Clinicians require simple quantitative tools for the detection of homogentisic acid in alkaptonuria patients, a rare inherited disorder of amino acid metabolism. In this study, we report a whole-cell biosensor for homogentisic acid to detect alkaptonuria disease through the expression of green fluorescence protein. The assay system utilizes a promoter sequence (hmgA) isolated from the Pseudomonas aeruginosa genome. To increase the sensitivity, the sensor module harboring phmgA::GFP was further transformed into various transposon mutants debilitated in steps involved in the metabolism of phenylalanine and tyrosine via homogentisic acid as a central intermediate. The proposed biosensor was further checked for analytical features such as sensitivity, selectivity, linearity, and precision for the quantification of homogentisic acid in spiked urine samples. The limit of detection for the developed biosensor was calculated to be 3.9 µM, which is comparable to that of the various analytical techniques currently in use. The sensor construct showed no interference from all of the amino acids and its homolog molecules. The accuracy and precision of the proposed biosensor were validated using high-performance liquid chromatography (HPLC) with satisfactory results.

Transcription factor based whole-cell biosensor for specific and sensitive detection of sodium dodecyl sulfate.

Extensive use of Sodium Dodecyl Sulfate (SDS) in households, agricultural operations, and industries is leading to its subsequent disposal in waterways. There is an apprehension of the adverse effect of such detergents on various living organisms. Thus, an efficient, specific, and simple detection method to monitor SDS reliably in the environment is needed. We used sdsB1 activator protein and SDS-responsive promoter of sdsA1 gene along with Green Fluorescent Protein (GFP) to construct a novel SDS biosensor in Pseudomonas aeruginosa chassis. The GFP intensity of the biosensor showed a linear relationship (R2 = 0.99) from 0.4 to 62.5 ppm of SDS with a detection limit of 0.1 ppm. This biosensor is highly specific for SDS and has minimal interference from other detergents, metals, and inorganic ions. The biosensor showed a satisfactory and reproducible recovery rate for the detection of SDS in real samples. Overall, this is a low cost, easy-to-use, selective, and reliable biosensor for monitoring SDS in the environment.

Occurrence, fates and potential treatment approaches for removal of viruses from wastewater: A review with emphasis on SARS-CoV-2.

The world is combating the emergence of Coronavirus disease 2019 (COVID-19) caused by novel coronavirus; severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Further, due to the presence of SARS-CoV-2 in sewage and stool samples, its transmission through water routes cannot be neglected. Thus, the efficient treatment of wastewater is a matter of utmost importance. The conventional wastewater treatment processes demonstrate a wide variability in absolute removal of viruses from wastewater, thereby posing a severe threat to human health and environment. The fate of SARS-CoV-2 in the wastewater treatment plants and its removal during various treatment stages remains unexplored and demands immediate attention; particularly, where treated effluent is utilised as reclaimed water. Consequently, understanding the prevalence of pathogenic viruses in untreated/treated waters and their removal techniques has become the topical issue of the scientific community. The key objective of the present study is to provide an insight into the distribution of viruses in wastewater, as well as the prevalence of SARS-CoV-2, and its possible transmission by the faecal-oral route. The review also gives a detailed account of the major waterborne and non-waterborne viruses, and environmental factors governing the survival of viruses. Furthermore, a comprehensive description of the potential methods (physical, chemical, and biological) for removal of viruses from wastewater has been presented. The present study also intends to analyse the research trends in microalgae-mediated virus removal and, inactivation. The review also addresses the UN SDG 'Clean Water and Sanitation' as it is aimed at providing pathogenically safe water for recycling purposes.

Cerebral malaria in a man with Plasmodium vivax mono-infection: a case report.

Cerebral malaria (CM) is a diffuse encephalopathy associated with coma and seizures commonly caused by Plasmodium falciparum (P. falciparum) in children with severe malaria. We present a case of a 19-year-old man with CM due to Plasmodium vivax (P. vivax) infection. Cerebrospinal fluid (CSF) was negative for Japanese B encephalitis, enterovirus, herpes simplex 1 and 2, varicella and mumps viruses as determined by real-time polymerase chain reaction (PCR). P. falciparum and P. vivax species were analysed by microscopy, immunochromatography and PCR assays and confirmed mono-infection of P. vivax in the patient's blood, and P. falciparum infection was established to be negative. The patient was discharged after intensive supportive care and antimalarial treatment (intravenous artesunate and oral doxycycline). We conclude that P. vivax infection is associated with CM, a life-threatening complication rarely seen in coastal districts of Karnataka. In endemic areas, the possibility of CM should be considered even with P. vivax infection.