Synergistic Consortium Research Articles

Isolation and screening of microplastics from Nodularia spumigena and Phaeodactylum tricornutum and determining the efficacy of biodegradation of microplastics obtained using the synergistic consortium

Microplastics are a form of plastics which are less than 5mm in size. They are the major threat for the environment and mankind, as they critically influence the food chain. The best way to extinct them from the environment is to degrade them in eco-friendly manner where biodegradation is the optimal therapy. Phytoplanktons play a vital role in phytoremediation. They are capable of in taking certain water pollutants such as micro plastics. Nodularia spumigena and Phaeodactylum tricornutum were screened and the microplastics were extracted using optimized enzymatic or alklanized digestion process and then dried. The micro plastics obtained were weighed and the concentration of microplastic present per gram of the sample was calculated. UV- VIS spectroscopy and FTIR instrumental analysis were used to analyse the digestion process. In order to formulate a synergistic consortium to degrade the thus obtained microplastic, plastic undergoing the natural degradation process from the dump yard was collected and crowded plate technique was performed and the strains involved in the degradation were thus isolated and identified and sequenced. Synergistic consortium was formulated and then incubated with the microplastics screened. The incubation period was of about 60 days. UV- Spectroscopic analysis was done on 0th day, 30th day and 60th day. The graph evidently illustrates the difference as the incubation time progresses. The peak depicts the microplastic degradation based on the turbidity. As the microplastic gets degraded, the growth of the organism in the consortium also gets limited because the microplastics are the only source of carbon.

Characterization and genetic determination of a newly isolated cotinine-degrading bacterium Terrabacter sp. strain cot-2 from synergistic consortium

Cotinine, the main metabolite of nicotine, was frequently detected in aquatic environment and pose a high risk to environmental safety and human health. Microbial degradation was considered as the most effective and environmental-friendly method for cotinine elimination. However, the microbial resources including bacterial consortia and pure cultures were limited. In this study, a synergistic consortium involved in cotinine degradation was successfully obtained and metagenomic analysis were investigated to reveal the key microorganisms responsible for cotinine and 6-hydroxy-3-succinoylpyridine (HSP) degradation, respectively. The genes encoding for cotinine hydroxylation (cotA1A2A3) and for HSP cleavage (hspB) were successfully amplified from the consortium C2 and functionally identified. Furthermore, a new cotinine-degrading pure culture strain, Terrabacter sp. cot-2, was isolated from the consortium C2. The degradation characterization and key intermediates of cotinine by strain cot-2 were analyzed. Then, the complete genome of strain cot-2 was determined and the gene cluster cot was demonstrated to be widely distributed in different environments. This study significantly broadened the knowledge on the degradation of the functional consortium and mechanism involved in cotinine biodegradation at molecular level, which also offering a promising solution to mitigate the impact of cotinine in aquatic ecosystems.

Development of an Antioxidant-Rich Sugar-Free Plantain Candy and Assessment of Its Shelf Life in a Flexible Laminate.

Candy is a popular confection worldwide, and it would be beneficial to society if it were converted into a source of antioxidant molecules to eliminate its adverse health effects. The amount of antioxidants available even in fruit candies is questionable due to the high thermal processing losses they undergo and the presence of various food additives. Plantains (Musa paradisiaca) are less known as good sources of biotherapeutic antioxidants, namely l-tryptophan, serotonin and melatonin, and consumption of this highly nutritious fruit is limited to underdeveloped and developing countries. The objectives of this study are: to develop a functional antioxidant-rich sugar-free plantain-based candy with valuable contents of the mentioned biomolecules in synergy; and to ensure its extended shelf life without compromising its physicochemical properties and functionality by wrapping it with a suitable packaging laminate. To accomplish the first objective, lyophilized plantain powder, sorbitol and mannitol were used as base materials with minimal additives under minimal processing conditions to reduce processing loss. Sensory, proximate, physicochemical and phytochemical properties, including the antioxidant synergy among the mentioned biomolecules of the developed candies were evaluated. For the second objective, the candies were enclosed in two different flexible packaging laminates and the optimal packaging was determined based on the microbiological safety and sensory appeal of the wrapped candies. Subsequently, the above-mentioned properties of the packaged (in the most suitable laminate) candies were evaluated at regular time intervals during storage for assessment of their shelf life. The candy had a characteristic flavour of plantain, uniform dark brown colour, rich mouthfeel, pleasant aroma, moderately hard texture and moderate sweetness, along with high antioxidant activity and considerable content of l-tryptophan, serotonin and melatonin (present as a synergistic consortium). During storage of the packaged candy under ambient conditions, it remained microbiologically safe for up to 56 days, and also maintained sensory attributes, antioxidant activity and synergy compared to the control candy. This newly developed semi-hard sugar-free candy with high antioxidant content, containing three important antioxidants, namely l-tryptophan, serotonin and melatonin, could be a good source of biotherapeutic molecules and a substitute for commercial candies consumed globally.

Parvimonas micra can translocate from the subgingival sulcus of the human oral cavity to colorectal adenocarcinoma.

Oral and intestinal samples from a cohort of 93 colorectal cancer (CRC) patients and 30 healthy controls (non-CRC) were collected for microbiome analysis. Saliva (28 non-CRC and 94 CRC), feces (30 non-CRC and 97 CRC), subgingival fluid (20 CRC), and tumor tissue samples (20 CRC) were used for 16S metabarcoding and/or RNA sequencing (RNAseq) approaches. A differential analysis of the abundance, performed with the ANCOM-BC package, adjusting the P-values by the Holm-Bonferroni method, revealed that Parvimonas was significantly over-represented in feces from CRC patients (P-value < 0.001) compared to healthy controls. A total of 11 Parvimonas micra isolates were obtained from the oral cavity and adenocarcinoma of CRC patients. Genome analysis identified a pair of isolates from the same patient that shared 99.2% identity, demonstrating that P. micra can translocate from the subgingival cavity to the gut. The data suggest that P. micra could migrate in a synergistic consortium with other periodontal bacteria. Metatranscriptomics confirmed that oral bacteria were more active in tumor than in non-neoplastic tissues. We suggest that P. micra could be considered as a CRC biomarker detected in non-invasive samples such as feces.

A Novel spice-antioxidant-based nano-vehicle as a putative green alternative of synthetic AChE inhibitor drugs

The present treatment for Alzheimer’s disease (AD) involves well known synthetic acetylcholine esterase (AChE) inhibitor drugs which besides having short duration of action also have deleterious impact on human health. Therefore, there is a need for natural plant-based biomolecule(s) with potential AChE inhibition activity (ies). The aim of the work is to design a spice-based nano-vehicle as a novel green alternative of synthetic AD drugs by nanoencapsulating a solvent-less supercritical CO2 extract of small cardamom seeds (SCE) having a synergistic consortium of five antioxidant molecules, using polyethylene glycol and emulsifiers, selected based on Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) analyses. Ellman’s assay and enzyme inhibition kinetics of the antioxidant molecules as well as the extract and its nanoliposomal formulation (SCE-NL) were performed, followed by rigorous molecular docking and dynamics studies using MM-PBSA and umbrella sampling. The antioxidants exhibited significant AChE inhibition in vitro, individually with 1, 8-cineole having the least IC50 value of 65.53 ± 0.05 µg/mL. . Although SCE-NL had higher IC50 value (575.67 ± 0.5 µg/mL) vis-à-vis that of rivastigmine (67.52 ± 0.02 µg/mL), it is safer for usage being ‘green’.The Lineweaver-Burk plots (Vmax ∼1.04 mM/min) revealed competitive mode(s) of inhibition of AChE with each of these antioxidants. Binding energy analyses suggested very good binding free energies and stable docking/binding complexes (between the antioxidants and AChE). This study has delivered a nanoliposomal vehicle of food antioxidants as a putative ‘green’ alternative of synthetic AChE inhibitor drugs. Communicated by Ramaswamy H. Sarma

Interspecies Metabolic Interactions in a Synergistic Consortium Drive Efficient Degradation of the Herbicide Bromoxynil Octanoate.

Microbial communities play vital roles in biogeochemical cycles, allowing biodegradation of a wide range of pollutants. Although many studies have shown the importance of interspecies interactions on activities of communities, fully elucidating the complex interactions in microbial communities is still challenging. Here, we isolated a consortium containing two bacterial strains (Acinetobacter sp. AG3 and Bacillus sp. R45), which could mineralize bromoxynil octanoate (BO) with higher efficiency than either strain individually. The BO degradation pathway by the synergistic consortium was elucidated, and interspecies interactions in the consortium were explored using genome-scale metabolic models (GSMMs). Modeling showed that growth and degradation enhancements were driven by metabolic interactions, such as syntrophic exchanges of small metabolites in the consortium. Besides, nutritional enhancers were predicted to improve BO degradation, which were tested experimentally. Overall, our results will enhance our understanding of microbial mineralization of BO by consortia and promote the application of microbial communities for bioremediation.

Melatonin‐rich, erucic acid‐lean nutraceutical supplements by microwave‐assisted solvent extraction of brown and yellow mustard seeds

The microwave-assisted solvent extraction (MASE) parameters optimized by Taguchi L9 (34) orthogonal array design to obtain extracts with a favorable combination of high melatonin-cum-low erucic acid content from Brassica juncea (BJ) and Brassica campestris (BC) seeds were: seed particle diameter of 500 μm for BJ and 420 μm for BC seeds, 1:10 sample to solvent (50% ethanolic water) ratio, 180 W microwave power, and 10 min extraction time. The yield of melatonin from BC was relatively higher owing to its greater protein and/or moisture content(s). Biochemical characterization confirmed a synergistic consortium of antioxidants in the extracts, viz. melatonin along with co-extracted ascorbic acid, limonene, and linalool. At the optimized condition, the actual power absorbed by the seed-solvent matrices (21.5 W in BJ-, 24.6 W in BC-MASE extracts) was lower compared to the microwave power applied (180 W) during MASE. Our studies affirm safe consumption of the extracts as melatonin-rich nutraceutical supplements. Novelty impact statement This work is of significance since melatonin which is commercially used as a drug has been extracted from an unconventional source, that is, from an oilseed crop for use as a food antioxidant. MASE has been conducted for the first time with green solvents for obtaining “melatonin rich-cum-erucic acid-lean” extracts of brown and yellow mustard seeds embracing natural “antioxidant synergy” within. The procedure established in this work could be easily scaled up to commercial level extraction of “a synergistic consortium of antioxidants” from other oilseeds (or allied food matrices).

PEGylated Nanoencapsulate of Melatonin-Rich Supercritical CO2 Extract of Yellow Mustard Seeds is an Authentic Lead for Type 2 Diabetes and Cholesterol Management

This study aimed to formulate melatonin (MT)-rich nanocapsulates from natural sources for mitigating hypercholesterolemia and type 2 diabetes. The in vivo hypocholesterolemic and antidiabetic efficacies of the MT-rich extract of yellow mustard (YM) seeds has already been established in our previous investigation. This outcome prompted us to formulate nanoliposomes of the aforesaid extract to enhance the said therapeutic efficacies. YM extract has been encapsulated in the form of nanoliposomes (YM-PN) and subjected to detailed functional characterization. YM-PN has been administered orally (550 mg/kg b.w. for 28 days) in Wistar albino rats and, post-sacrification their plasma MT level, fasting blood glucose (FBG) level, serum lipid profile and activities of key enzymes of glucose and cholesterol metabolic pathways were assayed. Functional characterization of YM-PN revealed PEGylated-o/w type-hydrophilic nanoliposomes which demonstrated prolonged and sustained release of MT (63.32% after 8 h). A synergistic consortium of antioxidants conferred strong antioxidant activity to YM-PN. Oral administration of YM-PN to diabetic and hypercholesterolemic rats restored their normal FBG and total cholesterol (TC) levels, respectively, with desirable up-down regulations of key enzymes of glucose and cholesterol homeostasis. Plasma MT levels also improved in these animals. The in vitro non-invasive model of iHOMA2 predicted that YM-PN increased hepatic insulin sensitivity and/ or glucose uptake in the gut in diabetic rats. The synergistic consortium of bioactives (including melatonin) in YM-PN is an authentic lead for natural product-based medicines in redressing hypercholesterolemia and type 2 diabetes.

Artificial Consortium of Three E. coli BL21 Strains with Synergistic Functional Modules for Complete Phenanthrene Degradation.

Polycyclic aromatic hydrocarbons (PAHs) are highly toxic and persistent organic pollutions that can accumulate in the environment. In this study, an aromatic ring cleavage module, a salicylic acid synthesis module, and a catechol metabolism module were respectively constructed in three Escherichia coli BL21 strains. Subsequently, the engineered strains were cocultured as an artificial consortium for the biodegradation of phenanthrene, a typical PHA. Single factor experiments and response surface methodology were used to identify the optimal degradation conditions, including an inoculation interval of 6 h, inoculation ratio of 1:1:1, and IPTG concentration of 2 mM. Under these conditions, the 7-day degradation ratio of 100 mg/L phenanthrene reached 72.67%. Moreover, the engineered Escherichia coli BL21 strains showed good phenanthrene degradation ability at substrate concentrations 10 mg/L up to 500 mg/L. Enzyme activity assays combined with gas chromatography-mass spectrometry measurements confirmed that the three engineered strains behaved as a synergistic consortium in the phenanthrene degradation process. Based on the analysis of the key metabolites, the engineered bacteria were supplemented at 7-day intervals in batches so that each engineered strain maintained its optimal degradation ability. The 21-day degradation ratio finally reached 90.66%, which was much higher than what was observed with simultaneous inoculation. These findings suggest that the three engineered strains with separate modules constructed in this study offer an attractive solution for removing PAHs from the environment.

A Synergistic Consortium Involved in rac-Dichlorprop Degradation as Revealed by DNA Stable Isotope Probing and Metagenomic Analysis.

rac-Dichlorprop, a commonly used phenoxyalkanoic acid herbicide, is frequently detected in environments and poses threats to environmental safety and human health. Microbial consortia are thought to play key roles in rac-dichlorprop degradation. However, the compositions of the microbial consortia involved in rac-dichlorprop degradation remain largely unknown. In this study, DNA stable isotope probing (SIP) and metagenomic analysis were integrated to reveal the key microbial consortium responsible for rac-dichlorprop degradation in a rac-dichlorprop-degrading enrichment. OTU340 (Sphingobium sp.) and OTU348 (Sphingopyxis sp.) were significantly enriched in the rac-[13C]dichlorprop-labeled heavy DNA fractions. A rac-dichlorprop degrader, Sphingobium sp. strain L3, was isolated from the enrichment by a traditional enrichment method but with additional supplementation of the antibiotic ciprofloxacin, which was instructed by metagenomic analysis of the associations between rac-dichlorprop degraders and antibiotic resistance genes. As revealed by functional profiling of the metagenomes of the heavy DNA, the genes rdpA and sdpA, involved in the initial degradation of the (R)- and (S)-enantiomers of dichlorprop, respectively, were mostly taxonomically assigned to Sphingobium species, indicating that Sphingopyxis species might harbor novel dichlorprop-degrading genes. In addition, taxonomically diverse bacterial genera such as Dyella, Sphingomonas, Pseudomonas, and Achromobacter were presumed to synergistically cooperate with the key degraders Sphingobium/Sphingopyxis for enhanced degradation of rac-dichlorprop. IMPORTANCE Understanding of the key microbial consortium involved in the degradation of the phenoxyalkanoic acid herbicide rac-dichlorprop is pivotal for design of synergistic consortia used for enhanced bioremediation of herbicide-contaminated sites. However, the composition of the microbial consortium and the interactions between community members during the biodegradation of rac-dichlorprop are unclear. In this study, DNA-SIP and metagenomic analysis were integrated to reveal that the metabolite 2,4-dichlorophenol degraders Dyella, Sphingomonas, Pseudomonas, and Achromobacter synergistically cooperated with the key degraders Sphingobium/Sphingopyxis for enhanced degradation of rac-dichlorprop. Our study provides new insights into the synergistic degradation of rac-dichlorprop at the community level and implies the existence of novel degrading genes for rac-dichlorprop in nature.

Synergistic consortium of beneficial microorganisms in rice rhizosphere promotes host defense to blight-causing Xanthomonas oryzae pv. oryzae.

Rice plants primed with beneficial microbes Bacillus amyloliquefaciens and Aspergillus spinulosporus with biocontrol potential against Xanthomonas oryzae pv. oryzae, provided protection from disease by reprogramming host defence response under pathogen challenge. Plant-beneficial microbe interactions taking place in the rhizosphere are widely used for growth promotion and mitigation of biotic stresses in plants. The present study aims to evaluate the defense network induced by beneficial microorganisms in the rice rhizosphere, and the three-way interaction involved upon inoculation with dreadful bacteria Xanthomonas oryzae pv. oryzae (Xoo). Differential expression of defense-related enzymes, proteins, and genes in rice variety Swarna primed with a microbial consortium of Bacillus amyloliquefaciens and Aspergillus spinulosporus were quantified in the presence and absence of Xoo. The time-based expression profile alterations in leaves under the five distinct treatments "(unprimed unchallenged, unprimed Xoo challenged, B. amyloliquefaciens primed and challenged, A. spinulosporus primed and challenged, B. amyloliquefaciens and A. spinulosporus consortium primed and challenged)" revealed differential early upregulation of SOD, PAL, PO, PPO activities and TPC content in beneficial microbes primed plants in comparison to unprimed challenged plants. The enhanced defense response in all the rice plants recruited with beneficial microbe was also reflected by reduced plant mortality and an increased plant dry biomass and chlorophyll content. Also, more than 550 protein spots were observed per gel by PD Quest software, a total of 55 differentially expressed protein spots were analysed used MALDI-TOF MS, out of which 48 spots were recognized with a significant score with direct or supporting roles in stress alleviation and disease resistance. qRT-PCR was carried out to compare the biochemical and proteomic data to mRNA levels. We conclude that protein biogenesis and alleviated resistance response may contribute to improved biotic stress adaptation. These results might accelerate the functional regulation of the Xoo-receptive proteins in the presence of beneficial rhizospheric microbes and their computation as promising molecular markers for superior disease management.

GMP-grade CD34+ selection from HLA-homozygous licensed cord blood units and short-term expansion under European ATMP regulations.

Based on a synergistic consortium, the cord blood (CB) bank Düsseldorf was responsible for the selection of HLA-homozygous (HLA-h) donors, contacting/re-consenting the mothers, Good Manufacturing Practice (GMP)-grade CD34+ enrichment, followed by short-term expansion of CD34+ cells and qualification of the resulting CD34+ population as advanced therapy medicinal product (ATMP)-starting material. Among 20639 licensed Düsseldorf cord blood units (CBUs), 139 potential HLA-h donors were identified with the most frequent 10 German haplotypes. 100% of the donors were contacted, and for 47·5%, consent was obtained. HLA-A, -B, -C, -DR, -DQ and -DP were determined by sequencing. Thawing/washing of the CBUs was performed in the presence of Volulyte/HSA with Sepax® , CD34+ selection by automated CliniMACS® -system (Miltenyi), expansion with qualified GMP-grade cytokines and media in the GMP facility. Here, we specify minimal criteria (≥5 x 105 viable CD34+ -count, ≥80% CD34+ -purity and ≥70% viability) and confirm that n=10 CB units (max storage time 16years) could be qualified for an ATMP starting material. The mean fold change expansion of isolated CD34+ cells at Day 3/4 (d3/4) was 3·38±3·02 with a mean purity of 86·90±10·38% and a high viability of 96·07±4·72%. As of March 2019, approval was obtained by the Bezirksregierung Düsseldorf for the GMP-compliant production. The production of HLA-homozygous expanded CD34+ cells from cryopreserved CB under European ATMP regulations presented here describes the successful clinical translation and implementation of a qualified manufacturing process. This approach considers the main obstacle of rejection of transplanted cells (due to the immunological HLA barrier) by preselection of HLA-homozygous transplants.

Nanoliposomes of Supercritical Carbon Dioxide Extract of Small Cardamom Seeds Redresses Type 2 Diabetes and Hypercholesterolemia.

In our previous investigation, oral administration of 1,8- cineole-rich supercritical carbon dioxide extract of small cardamom seeds in Wistar albino rats resulted in achieving normal fasting blood glucose (FBG) and serum cholesterol levels. The objective of this study was to further protect the aforesaid extract and to enhance its in vivo therapeutic efficacies in redressing type 2 diabetes and hypercholesterolemia, by encapsulating it as nanoliposomes. Patents related to nanoliposomes have been revised thoroughly. PEGylated nanoliposomes of the aforesaid extract were formulated using soya phosphatidylcholine and Tween 80 by probe-sonication. These nanoliposomes were subjected to in vitro characterizations and were orally administered to Wistar albino rats at three different doses viz. 550, 175 and 55 mg/kg b.w. for detailed investigation of their antidiabetic and hypocholesterolemic efficacies. FT-IR, DSC and XRD analyses, HLB value (16), entrapment efficiency (84%) and release kinetics (obeying Higuchi model) revealed that the nanoliposomes were o/w type and were hydrophilic. They exhibited appreciable in vitro antioxidant potency (59% DPPH scavenging activity) owing to a synergistic consortium of antioxidants present therein. Oral administration of the liposomes in rats at 550 mg/kg b.w. could restore their normal FBG levels and serum lipid profiles on day 35, with desirable up-down regulations of related key enzymes. The iHOMA2 model could successfully predict the effects of nanoliposomes on insulin sensitivity and glucose uptake in rat liver and brain, respectively. Nanoliposome of 1,8-cineole rich extract of small cardamom seeds is a new biotherapeutic in redressing type 2 diabetes and hypercholesterolemia.

Metabolomic Analysis of Cooperative Adaptation between Co-Cultured Bacillus cereus and Ketogulonicigenium vulgare

The cooperative adaptation of subcultivated Bacillus cereus and Ketogulonicigenium vulgare significantly increased the productivity of 2-keto-L-gulonic acid, the precursor of vitamin C. The mechanism of cooperative adaptation of the serial subcultivated B. cereus and K. vulgare was investigated in this study by culturing the two strains orthogonally on agar plates. It was found that the swarming distance of B. cereus along the trace of K. vulgare on the plate decreased after 150 days' subcultivation. Metabolomic analysis on these co-cultured B. cereus and K. vulgare strains showed that their cooperative adaptation was accomplished by three key events: (i) the ability of nutrients (e.g., amino acids and purines) searching and intaking, and proteins biosynthesis is increased in the evolved B. cereus; (ii) the capability of protein degradation and amino acids transportation is enhanced in evolved K. vulgare; (iii) the evolved B. cereus was found to provide more nutrients (mostly amino acids and purines) to K. vulgare, thus strengthening the oxidation and energy generation of K. vulgare. Our results provided novel insights into the systems-level understanding of the cooperative adaptation between strains in synergistic consortium.

Computational intelligence and visual computing: an emerging technology for software engineering

The discipline of Software Engineering is abstract and complex with all its endeavors being cast in a knowledge-intensive environment. It is not surprising that there have been a number of important initiatives that have attempted to address a burning need for solid development tools and comprehensive environments supporting an in-depth analysis. The objective of this study is to discuss a role of Computational Intelligence (CI) and visual computing being viewed as a sound methodological and algorithmic environment for knowledge-oriented Software Engineering. The CI itself is regarded as a synergistic consortium of granular computing (including fuzzy sets) promoting abstraction, neurocomputing supporting various learning schemes and evolutionary computing providing important faculties of global optimization. By its very nature, CI embraces a diversity of design paradigms; in particular it promotes a top-down approach (when exploiting fuzzy sets first and afterwards working in the neural network environment) or bottom-up style (where these two technologies are used in a reverse order). Visual computing is inherently associated with CI: it is human-centric where fuzzy sets make visualization activities feasible. Fuzzy sets are treated as a graphic means of accepting information from users. They are regarded as a vehicle used to visualize results in a linguistic manner. Software Engineering and CI are highly compatible: they are knowledge-intensive, human-oriented, and have to deal with various manifestations of the abstract world of software constructs and thought processes. This multifaceted conceptual compatibility is a prerequisite for the development of vital synergistic links that bring the technology of CI into Software Engineering. The symbiosis accrues considerable benefits for both technologies by posing new categories of challenging and highly stimulating problems. The facet of visual computing is essential in handling of software processes and software products. The intent of this study is to provide a general overview of this new development in Software Engineering. In particular, we highlight a number of selected and most visible trends occurring at the junction of CI and Software Engineering. Furthermore we discuss several specific applications of the technology of CI to software cost estimation, analysis of software measures and neural models of software quality.