Genomic, functional, and metabolic enhancements in multidrug-resistant Enterobacter bugandensis facilitating its persistence and succession in the International Space Station.

BACKGROUND: The International Space Station (ISS) stands as a testament to human achievement in space exploration. Despite its highly controlled environment, characterised by microgravity, increased CO 2 levels, and elevated solar radiation, microorganisms occupy a unique niche. These microbial inhabitants play a significant role in influencing the health and well-being of astronauts on board. One microorganism of particular interest in our study is Enterobacter bugandensis, primarily found in clinical specimens including the human gastrointestinal tract, and also reported to possess pathogenic traits, leading to a plethora of infections. RESULTS: Distinct from their Earth counterparts, ISS E. bugandensis strains have exhibited resistance mechanisms that categorise them within the ESKAPE pathogen group, a collection of pathogens recognised for their formidable resistance to antimicrobial treatments. During the 2-year Microbial Tracking 1 mission, 13 strains of multidrug-resistant E. bugandensis were isolated from various locations within the ISS. We have carried out a comprehensive study to understand the genomic intricacies of ISS-derived E. bugandensis in comparison to terrestrial strains, with a keen focus on those associated with clinical infections. We unravel the evolutionary trajectories of pivotal genes, especially those contributing to functional adaptations and potential antimicrobial resistance. A hypothesis central to our study was that the singular nature of the stresses of the space environment, distinct from any on Earth, could be driving these genomic adaptations. Extending our investigation, we meticulously mapped the prevalence and distribution of E. bugandensis across the ISS over time. This temporal analysis provided insights into the persistence, succession, and potential patterns of colonisation of E. bugandensis in space. Furthermore, by leveraging advanced analytical techniques, including metabolic modelling, we delved into the coexisting microbial communities alongside E. bugandensis in the ISS across multiple missions and spatial locations. This exploration revealed intricate microbial interactions, offering a window into the microbial ecosystem dynamics within the ISS. CONCLUSIONS: Our comprehensive analysis illuminated not only the ways these interactions sculpt microbial diversity but also the factors that might contribute to the potential dominance and succession of E. bugandensis within the ISS environment. The implications of these findings are twofold. Firstly, they shed light on microbial behaviour, adaptation, and evolution in extreme, isolated environments. Secondly, they underscore the need for robust preventive measures, ensuring the health and safety of astronauts by mitigating risks associated with potential pathogenic threats. Video Abstract.

Tuning Electrostatic Interactions To Control Orientation of GFP Protein Adsorption on Silica Surface.

The adsorption of green fluorescent protein (GFP) on silica surfaces has been the subject of growing interest due to its potential applications in various fields, including biotechnology and biomedicine. In this study, we used all-atom molecular dynamics simulations to investigate the charge-driven adsorption of wild type GFP and its supercharged variants on silica surfaces. The results showed that the positively charged variant of GFP adsorbed on the negatively charged silica surface with minimal loss in its secondary structure. Further studies were conducted to understand the role of surface charge distribution on two other positively charged variants of GFP, and the results showed that the orientation of GFP on silica can be easily tuned by careful mutations of the charged amino acid residues on the GFP. This study provides valuable molecular insights into the role of electrostatic-driven adsorption of GFP and highlights the importance of charge interactions in the adsorption process.

Enhancement in Capacitance of Ionic Type of EAP-Based Strain Sensors.

This paper aims to enhance the capacitance of electroactive polymer (EAP)-based strain sensors. The enhancement in capacitance was achieved by using a free-standing stretchable polymer film while introducing conducting polymer to fabricate a hybrid dielectric film with controlled conductivity. In this work, styrene-ethylene-butylene-styrene (SEBS) rubber was used as the base material, and dodecyl benzene sulfonate anion (DBSA)-doped polyaniline (PANI) was used as filler to fabricate a hybrid composite conducting film. The maleic anhydride group of the SEBS Rubber and DBSA, the anion of the polyaniline dopant, make a very stable dispersion in Toluene and form a free-standing stretchable film by solution casting. DBSA-doped polyaniline increased the conductivity and dielectric constant of the dielectric film, resulting in a significant enhancement in the capacitance of the EAP-based strain sensor. The sensor presented in this article exhibits capacitance values ranging from 24.7 to 100 µF for strain levels ranging from 0 to 100%, and sensitivity was measured 3 at 100% strain level.

Genomic analysis reveals the presence of emerging pathogenic Klebsiella lineages aboard the International Space Station.

IMPORTANCE: The International Space Station (ISS) is a unique, hermetically sealed environment, subject to environmental pressures not encountered on Earth, including microgravity and radiation (cosmic ionising/UV). While bacteria's adaptability during spaceflight remains elusive, recent research suggests that it may be species and even clone-specific. Considering the documented spaceflight-induced suppression of the human immune system, a deper understanding of the genomics of potential human pathogens in space could shed light on species and lineages of medical astromicrobiological significance. In this study, we used hybrid assembly methods and comparative genomics to deliver a comprehensive genomic characterization of 10 Klebsiella isolates retrieved from the ISS. Our analysis unveiled that Klebsiella quasipneumoniae ST138 demonstrates both spatial and temporal persistence aboard the ISS, showing evidence of genomic divergence from its Earth-based ST138 lineage. Moreover, we characterized plasmids from Klebsiella species of ISS origin, which harbored genes for disinfectant resistance and enhanced thermotolerance, suggestin possible adaptive advantages. Furthermore, we identified a mobile genetic element containing a hypervirulence-associated locus belonging to a Klebsiella pneumoniae isolate of the "high-risk" ST101 clone. Our work provides insights into the adaptability and persistence of Klebsiella species during spaceflight, highlighting the importance of understanding the dynamics of potential pathogenic bacteria in such environments.

Fate of pesticides in agricultural runoff treatment systems: Occurrence, impacts and technological progress.

The levels of pesticides in air, water, and soil are gradually increasing due to its inappropriate management. In particular, agricultural runoff inflicts the damages on the ecosystem and human health at massive scale. Present study summarizes 70 studies in which investigations on removal or treatment of pesticides/insecticides/herbicides are reported. A bibliometric analysis was also done to understand the recent research trends through the analysis of 2218 publications. The specific objectives of this study are as follows: i) to inventorize the characteristics details of agriculture runoff and analyzing the occurrence and impacts of pesticides, ii) analyzing the role and interaction of pesticides in different environmental segments, iii) investigating the fate of pesticides in agriculture runoff treatment systems, iv) summarizing the experiences and findings of most commonly technology deployed for pesticides remediation in agriculture runoff including target pesticide(s), specifications, configuration of technological intervention. Among the reported technologies for pesticide treatment in agriculture runoff, constructed wetland was at the top followed by algal or photobioreactor. Among various advanced oxidation processes, photo Fenton method is mainly used for pesticides remediation such as triazine, methyl parathion, fenuron and diuron. Algal bioreactors are extensively used for a wide range of pesticides treatment including 2,4-Dichlorophenoxyacetic acid, 2-methyl-4-chlorophenoxyacetic acid, alachlor, diuron, chlorpyrifos, endosulfan, and imidacloprid; especially at lower hydraulic retention time of 2-6 h. This study highlights that hybrid approaches can offers potential opportunities for effective removal of pesticides in a more viable manner.

Hydrophobicity─A Single Parameter for the Accurate Prediction of Disordered Regions in Proteins.

The prediction of disordered regions in proteins is crucial for understanding their functions, dynamics, and interactions. Intrinsically disordered proteins (IDPs) play a key role in many biological processes like cell signaling, recognition, and regulation, but experimentally determining these regions can be challenging due to their high mobility. To address this challenge, we present an algorithm called HydroDisPred (HDP). HDP uses a single parameter, the fraction of hydrophobicity (λ) in each segment of the protein, to accurately predict disordered regions. The algorithm was validated using experimental data from the DisProt database and was found to be on par and, in some cases, more effective than the existing algorithms. HDP is a simple and effective method for identifying disordered regions in proteins, and its prediction is not affected by the availability of training data, unlike other ML approaches. The application is housed in the web server and can be accessed through the URL https://proseqanalyser.iitgn.ac.in/hydrodispred/.

Medical Astro-Microbiology: Current Role and Future Challenges.

The second and third decades of the twenty-first century are marked by a flourishing of space technology which may soon realise human aspirations of a permanent multiplanetary presence. The prevention, control and management of infection with microbial pathogens is likely to play a key role in how successful human space aspirations will become. This review considers the emerging field of medical astro-microbiology. It examines the current evidence regarding the risk of infection during spaceflight via host susceptibility, alterations to the host's microbiome as well as exposure to other crew members and spacecraft's microbiomes. It also considers the relevance of the hygiene hypothesis in this regard. It then reviews the current evidence related to infection risk associated with microbial adaptability in spaceflight conditions. There is a particular focus on the International Space Station (ISS), as one of the only two  crewed objects in low Earth orbit. It discusses the effects of spaceflight related stressors on viruses and the infection risks associated with latent viral reactivation and increased viral shedding during spaceflight. It then examines the effects of the same stressors on bacteria, particularly in relation to changes in virulence and drug resistance. It also considers our current understanding of fungal adaptability in spaceflight. The global public health and environmental risks associated with a possible re-introduction to Earth of invasive species are also briefly discussed. Finally, this review examines the largely unknown microbiology and infection implications of celestial body habitation with an emphasis placed on Mars. Overall, this review summarises much of our current understanding of medical astro-microbiology and identifies significant knowledge gaps.

Microbial Technologies in Waste Management, Energy Generation and Climate Change: Implications on Earth and Space.

Microbes are important decomposers of organic waste. By decomposing organic waste and using it for their growth, microbes play an important role in maintaining ecosystem's carbon and nitrogen cycles. An ecosystem's microbial shift may disturb it's carbon/nitrogen cycle as a result of any climate change or humanitarian factors, but heat produced by various instruments and greenhouse gases contribute significantly to global warming which in turn may be related to microbial shift of ecosystems. To reduce greenhouse gas emissions and global warming, innovative clean energy production methods must be employed to develop fuels with minimal greenhouse effect. Biofuels, such as bioethanol, provide clean energy with less carbon dioxide emissions. For the production of bioethanol, it is always recommended to use microbes that are capable of decomposing complex organic matter (cellulose, lignin, hemicellulose). Some microbes can efficiently decompose complex organic matter due to the presence of genetic machinery that produces cellulases and ß-glucosidase. The membrane transporters are also important for microbes in uptake of simple sugars for metabolism and ethanol production. Microbial technologies are addressing the future needs for not only organic waste management but also clean energy/bioethanol production. However, the role of these technologies on space missions and extraterrestrial settings needs to be explored to improve long term space missions.

Understanding the helical stability of charged peptides.

Cationic helical peptides play a crucial role in applications such as anti-microbial and anticancer activity. The activity of these peptides directly correlates with their helicity. In this study, we have performed extensive all-atom molecular dynamics simulations of 25 Lysine-Leucine co-polypeptide sequences of varying charge density ( λ ) and patterns. Our findings showed that, an increase in the charge density on the peptide leads to a gradual decrease in the helicity up to a critical charge density λ c . Beyond λ c , a complete helix to coil transition was observed. The decrease in the helicity is correlated with the increased number of water molecules in first solvation shell, solvent-exposed surface area, and a higher value of the radius of gyration of the peptide.

Artificial intelligence and machine learning-based monitoring and design of biological wastewater treatment systems.

Artificial intelligence (AI) and machine learning (ML) are currently used in several areas. The applications of AI and ML based models are also reported for monitoring and design of biological wastewater treatment systems (WWTS). The available information is reviewed and presented in terms of bibliometric analysis, model's description, specific applications, and major findings for investigated WWTS. Among the applied models, artificial neural network (ANN), fuzzy logic (FL) algorithms, random forest (RF), and long short-term memory (LSTM) were predominantly used in the biological wastewater treatment. These models are tested by predictive control of effluent parameters such as biological oxygen demand (BOD), chemical oxygen demand (COD), nutrient parameters, solids, and metallic substances. Following model performance indicators were mainly used for the accuracy analysis in most of the studies: root mean squared error (RMSE), mean square error (MSE), and determination coefficient (DC). Besides, outcomes of various models are also summarized in this study.

First complete genome of a multidrug-resistant strain of the novel human pathogen Kalamiella piersonii (GABEKP28) identified in human saliva.

OBJECTIVES: Kalamiella piersonii is a newly identified bacterial species, first isolated from surfaces of the International Space Station (ISS). It also appears as a novel human pathogen reported to be implicated in bacteremia and kidney stone disease. Here, we report the first complete genome of a multidrug-resistant strain of K. piersonii (GABEKP28), isolated from the saliva of a patient with treatment-resistant schizophrenia (TRS), to determine the mobile genetic elements (MGEs), antibiotic resistance genes (ARGs), and virulence factors (VFs) harboured by such a strain of this novel species. METHODS: Whole-genome sequencing was performed using DNABSEQ (PE150) and Nanopore MinION platforms. Hybrid assembly was conducted using Unicycler v0.5.0. Genome assembly quality was verified using QUAST v5.0.2. The assembly was annotated using PROKKA v1.14.5. ARGs and VFs were identified using Abricate v1.0.0. RESULTS: K. piersonii strain GABEKP28 was classified as multidrug-resistant while also carrying plasmidic genetic determinants associated with a hypervirulent phenotype. The complete genome size is 3 881 479 bp and has a guanine-cytosine content of 57.76% while it encodes for 3 525 chromosome coding sequences. The strain was also identified to carry three plasmids of 513 647 bp, 261 771 bp, and 106 029 bp, respectively. CONCLUSIONS: K.piersonii GABEKP28 is the first complete genome of this species to be submitted to GenBank and only the second to be sequenced from a human host. The whole-genome sequencing data with multiple plasmids, ARGs, and VFs will aid in understanding the pathogenicity, evolution, and phylogeny of this novel opportunistic pathogen.

Metabolic modeling of the International Space Station microbiome reveals key microbial interactions.

BACKGROUND: Recent studies have provided insights into the persistence and succession of microbes aboard the International Space Station (ISS), notably the dominance of Klebsiella pneumoniae. However, the interactions between the various microbes aboard the ISS and how they shape the microbiome remain to be clearly understood. In this study, we apply a computational approach to predict possible metabolic interactions in the ISS microbiome and shed further light on its organization. RESULTS: Through a combination of a systems-based graph-theoretical approach, and a constraint-based community metabolic modeling approach, we demonstrated several key interactions in the ISS microbiome. These complementary approaches provided insights into the metabolic interactions and dependencies present amongst various microbes in a community, highlighting key interactions and keystone species. Our results showed that the presence of K. pneumoniae is beneficial to many other microorganisms it coexists with, notably those from the Pantoea genus. Species belonging to the Enterobacteriaceae family were often found to be the most beneficial for the survival of other microorganisms in the ISS microbiome. However, K. pneumoniae was found to exhibit parasitic and amensalistic interactions with Aspergillus and Penicillium species, respectively. To prove this metabolic prediction, K. pneumoniae and Aspergillus fumigatus were co-cultured under normal and simulated microgravity, where K. pneumoniae cells showed parasitic characteristics to the fungus. The electron micrography revealed that the presence of K. pneumoniae compromised the morphology of fungal conidia and degenerated its biofilm-forming structures. CONCLUSION: Our study underscores the importance of K. pneumoniae in the ISS, and its potential positive and negative interactions with other microbes, including potential pathogens. This integrated modeling approach, combined with experiments, demonstrates the potential for understanding the organization of other such microbiomes, unravelling key organisms and their interdependencies. Video Abstract.

The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly Cleanrooms.

Ensuring biological cleanliness while assembling and launching spacecraft is critical for robotic exploration of the solar system. To date, when preventing forward contamination of other celestial bodies, NASA Planetary Protection policies have focused on endospore-forming bacteria while fungi were neglected. In this study, for the first time the mycobiome of two spacecraft assembly facilities at Jet Propulsion Laboratory (JPL) and Kennedy Space Center (KSC) was assessed using both cultivation and sequencing techniques. To facilitate enumeration of viable fungal populations and downstream molecular analyses, collected samples were first treated with chloramphenicol for 24 h and then with propidium monoazide (PMA). Among cultivable fungi, 28 distinct species were observed, 16 at JPL and 16 at KSC facilities, while 13 isolates were potentially novel species. Only four isolated species Aureobasidium melanogenum, Penicillium fuscoglaucum, Penicillium decumbens, and Zalaria obscura were present in both cleanroom facilities, which suggests that mycobiomes differ significantly between distant locations. To better visualize the biogeography of all isolated strains the network analysis was undertaken and confirmed higher abundance of Malassezia globosa and Cyberlindnera jadinii. When amplicon sequencing was performed, JPL-SAF and KSC-PHSF showed differing mycobiomes. Metagenomic fungal reads were dominated by Ascomycota (91%) and Basidiomycota (7.15%). Similar to amplicon sequencing, the number of fungal reads changed following antibiotic treatment in both cleanrooms; however, the opposite trends were observed. Alas, treatment with the antibiotic did not allow for definitive ascribing changes observed in fungal populations between treated and untreated samples in both cleanrooms. Rather, these substantial differences in fungal abundance might be attributed to several factors, including the geographical location, climate and the in-house cleaning procedures used to maintain the cleanrooms. This study is a first step in characterizing cultivable and viable fungal populations in cleanrooms to assess fungal potential as biocontaminants during interplanetary explorations. The outcomes of this and future studies could be implemented in other cleanrooms that require to reduce microbial burden, like intensive care units, operating rooms, or cleanrooms in the semiconducting and pharmaceutical industries.

Human Health during Space Travel: State-of-the-Art Review.

The field of human space travel is in the midst of a dramatic revolution. Upcoming missions are looking to push the boundaries of space travel, with plans to travel for longer distances and durations than ever before. Both the National Aeronautics and Space Administration (NASA) and several commercial space companies (e.g., Blue Origin, SpaceX, Virgin Galactic) have already started the process of preparing for long-distance, long-duration space exploration and currently plan to explore inner solar planets (e.g., Mars) by the 2030s. With the emergence of space tourism, space travel has materialized as a potential new, exciting frontier of business, hospitality, medicine, and technology in the coming years. However, current evidence regarding human health in space is very limited, particularly pertaining to short-term and long-term space travel. This review synthesizes developments across the continuum of space health including prior studies and unpublished data from NASA related to each individual organ system, and medical screening prior to space travel. We categorized the extraterrestrial environment into exogenous (e.g., space radiation and microgravity) and endogenous processes (e.g., alteration of humans' natural circadian rhythm and mental health due to confinement, isolation, immobilization, and lack of social interaction) and their various effects on human health. The aim of this review is to explore the potential health challenges associated with space travel and how they may be overcome in order to enable new paradigms for space health, as well as the use of emerging Artificial Intelligence based (AI) technology to propel future space health research.

Investigations on air quality of a critically polluted industrial city using multivariate statistical methods: Way forward for future sustainability.

Industrially developed cities affect public health, and can directly cause inconvenience to the nearby societies especially due to their associated air pollution. In this context, the present study was conducted in Jharsuguda district of Odisha state (India), which is a well-known worldwide hub of industrial clusters. The study area is having mainly medium to large scale industries which makes it prone to poor air quality. A total of twelve air pollutants, namely, PM10, PM2.5, SO2, NO2, CO, O3, NH3, and heavy metals (Cu, Mn, Ni, Pb, Zn) were monitored during winter season, at the 16 locations of study area. The air quality data was further assessed using multivariate analysis, and the obtained information was presented using histogram plots, box plots, cluster analysis, principal component analysis (PCA), analysis of variance (ANOVA) analysis, and air quality index (AQI). The statistical analysis results revealed that PM10 and PM2.5 levels exceeded the permissible limits of study area, ∼40 and 30% of sampling times, respectively. Contrary, values of other pollution parameters were observed to be well within the permissible limits. The cluster analysis distinguishingly summarized the monitoring data into four clusters types, named as severely polluted, moderately polluted, satisfactory, and fine. The PCA analysis of monitored data resulted in identification of prominent emission sources of analyzed pollutants. These sources were mainly found to be associated with coal burning in power plants, agricultural activities, vehicular emissions, and mining activities. The minimum AQI was observed as 87 at Orient (mine no. 4) and Kinjirma which is under satisfactory category, whereas maximum AQI was observed at Bhedabahal with a value of 132 which is under moderate category. Overall, the results of this study indicated that the air pollution of industrial areas must be evaluated thoroughly on regular basis, considering the sustainability of societies and expanding industries.

Understanding the role of hydrophobic patches in protein disaggregation.

Protein folding is a very complex process and, so far, the mechanism of folding still intrigues the research community. Despite a large conformational space available (O(1047) for a 100 amino acid residue), most proteins fold into their native state within a very short time. While small proteins fold relatively fast (a few microseconds) large globular proteins may take as long as several milliseconds to fold. During the folding process, the protein synthesized in the ribosome is exposed to the crowded environment of the cell and is easily prone to misfolding and aggregation due to interactions with other proteins or biomacromolecules present within the cell. These large proteins, therefore, rely on chaperones for their folding and repair. Chaperones are known to have hydrophobic patchy domains that play a crucial role in shielding the protein against misfolding and disaggregation of aggregated proteins. In the current article, Monte Carlo simulations carried out in the framework of the hydrophobic-polar (H-P) lattice model indicate that hydrophobic patchy domains drastically reduce the inter-protein interactions and are efficient in disaggregating proteins. The effectiveness of the disaggregation depends on the size and distribution of these patches on the surface and also on the strength of the interaction between the protein and the surface. Further, our results indicate that when the patch is complementary to the exposed hydrophobic patch of the protein, protein disaggregation is accompanied by stabilization of the protein even relative to its bulk behavior due to favorable protein-surface interactions. We believe that these findings shed light on the role of the class of chaperones known as heat shock proteins (Hsps) on protein disaggregation and refolding.

Advances in space microbiology.

Microbial research in space is being conducted for almost 50 years now. The closed system of the International Space Station (ISS) has acted as a microbial observatory for the past 10 years, conducting research on adaptation and survivability of microorganisms exposed to space conditions. This adaptation can be either beneficial or detrimental to crew members and spacecraft. Therefore, it becomes crucial to identify the impact of two primary stress conditions, namely, radiation and microgravity, on microbial life aboard the ISS. Elucidating the mechanistic basis of microbial adaptation to space conditions aids in the development of countermeasures against their potentially detrimental effects and allows us to harness their biotechnologically important properties. Several microbial processes have been studied, either in spaceflight or using devices that can simulate space conditions. However, at present, research is limited to only a few microorganisms, and extensive research on biotechnologically important microorganisms is required to make long-term space missions self-sustainable.

Draft Genome Sequences of Aspergillus and Penicillium Species Isolated from the International Space Station and Crew Resupply Vehicle Capsule.

The draft whole-genome sequences (WGS) of 30 fungal strains isolated from the International Space Station and belonging to the Penicillium and Aspergillus genera were assembled. The WGS will allow for detailed genomic characterization to determine the possible applications and importance for space and biotechnological industries.

Organizational resilience and social-economic sustainability: COVID-19 perspective.

COVID-19 has affected the global economy like no other crisis in the history of mankind. It forced worldwide lockdown and economic shutdown to the point from where the recovery process has been very difficult. It has affected demand, supply, production and consumption in such a way that the entire economic development cycle has gone to its lowest levels. COVID-19 has also affected the social and economic sustainability structure which has led from one crisis to another and the developing countries have been the worst hit. Economic crisis resulted in unemployment leading to labour migrations, inevitable casualties and rising poverty etc. However, at a certain level, a few industries and organizations have shown resilience with better anticipation and survivability which may lead them to a quicker recovery. The current study aims at presenting a holistic view of organizational resilience which leads to the overall sustainable development. The study considers three aspects of organizational resilience as crisis anticipation, organizational robustness and recoverability. It assesses the impact of the aspects of resilience on social sustainability and economic sustainability. The study uses empirical analysis of primary data which is analysed to verify the hypothesized relationships by using a structural equation modelling approach. The study finds out that predicting the crisis and disruptions, building robustness and recoverability have a positive effect on both the social and economic aspects of sustainability. Findings of the study have their practical implications for industry, researchers and society.

Greenhouse gases emission control in WWTS via potential operational strategies: A critical review.

Greenhouse gases (GHGs; particularly, CO2, CH4, and N2O) emission from wastewater treatment systems (WWTS) is one of the inevitable concerns for sustainable development. This indicator is directly linked with the carbon footprint and potential impacts of WWTS on climate change. In this view, various modeling, design, and operational tools have been introduced to mitigate the WWTS associated GHGs, at regional and global scales. In this study, authors have critically reviewed the selected potential operational control strategies for GHGs emission, particularly emitted from the operational stages of biological WWTS. The investigated operational control strategies and/or treatment configurations included intermittent aeration, varying dissolved oxygen, enhanced sludge retention time, coupled aerobic-anoxic nitrous decomposition operation, and microalgae integrated treatment process. Based on this analysis and considering the trade-off between treatment performance of WWTS and GHGs control, an integrated framework is also proposed for existing and upcoming WWTS. The findings of this study and proposed framework will play an instrumental role in mitigating the GHGs at various operational stages of WWTS. Future research works in this direction can lead to a better understanding of investigated operational GHGs emission control strategies in WWTS.