Journal of Experimental Biology and Agricultural Sciences

Production, characterization, and applications of a novel thermo-acidophilic L-asparaginase of Pseudomonas aeruginosa CSPS4

Vinay Kumar — 2024-03-15

In present investigation, a potential L-asparaginase-producing bacterial isolate, Pseudomonas aeruginosa CSPS4, has been explored to enhance the production and purification of the asparaginase enzyme. Production of L-asparaginase is enhanced using the 'one variable at a time approach (OVAT)'. In Placket Burman (PB) analysis, pH, sucrose, and temperature significantly influence L-asparaginase production. Thereafter, L-asparaginase enzyme was recovered from culture broth using fractional precipitation with chilled acetone. The partially purified L-asparaginase showed a molecular weight of ~35 KDa on SDS-PAGE. L-asparaginase was characterized as a thermo-acidophilic enzyme exhibiting optimum pH and temperature of 6.0 and 60 °C, respectively. These characteristics render this enzyme novel from other available asparaginases of Pseudomonas spp. L-asparaginase activity remained unaffected by different modulators. L-asparaginase of this investigation was successfully employed for acrylamide degradation in commercial fried potato chips, establishing its applicability in food industries.

FIRST REPORT ON TRUFFLE-INHABITING FUNGI AND METAGENOMIC COMMUNITIES OF TUBER AESTIVUM COLLECTED IN RUSSIA

Ekaterina V. Malygina — 2024-03-15

Truffles are one of the least studied groups of fungi in terms of their biological and biotechnological aspects. This study aimed to isolate truffle-inhabiting fungi and assess the metagenomic communities of the most common Russian summer truffle, Tuber aestivum. This study is the first to characterize the biodiversity of prokaryotic and eukaryotic organisms living in the truffle T. aestivum using molecular analysis and sequencing. Plant pathogens involved in a symbiotic relationship with truffles were identified by sequencing the hypervariable fragments of the 16S rRNA and 18S rRNA genes. In addition, some strains of fungal symbionts and likely pathogens were isolated and recognized for the first time from the truffles. This study also compared and characterized the general diversity and distribution of microbial taxa of T. aestivum collected in Russia and Europe. The results revealed that the Russian and European truffle study materials demonstrated high similarity. In addition to the truffles, representatives of bacteria, fungi, and protists were found in the fruiting bodies. Many of these prokaryotic and eukaryotic species inhabiting truffles might influence them, help them form mycorrhizae with trees, and regulate biological processes. Thus, truffles are interesting and promising sources for modern biotechnological and agricultural studies.

Role of Probiotic Strain Lactobacillus acidophilus in the Reversal of Gut Dysbiosis Induced Brain Cognitive Decline

Murugan Mukilan — 2024-03-15

In the central nervous system, bidirectional communication between the brain and gut results in memory formation due to synaptic plasticity changes. During a healthy state, oral balanced microflora plays a pivotal role in memory formation by inhibiting the enterotoxin level produced by infectious pathogens. In disease conditions, beneficial microbial dysbiosis may result in excess enterotoxin production. Further, excess enterotoxin secretion prevents beneficial bacteria's proliferation and impairs neurotransmitter precursor compounds' transport to the brain. Blockade of neurotransmitter precursor compounds may result in the development of memory loss. The present study stated the role of Lactobacillus acidophilus in recovering memory loss. Reversal of cognitive impairment is shown with the help of a three-step behavioural analysis, which consists of one pre-infusive behavioural analysis and two post-infusive behavioural analyses (phase 1 and 2). The pre-infusive analysis showed no cognitive impairment in an assimilated environment without any infusions. After oral microbial infusions, phase 1 of post-infusive behavioural analysis showed the presence of cognitive impairment in the experimental groups who received oral infusions. Formed cognitive impairment is reverted with the help of L. acidophilus oral infusion in phase 2 of post-infusive analysis. Comparative three-step behavioural analysis proved that Pseudomonas aeuroginosa induced cognitive impairment may revert to normal conditions with the help of L. acidophilus. The outcome of the present study proves that cognitive impairment developed due to poor oral hygiene can be treated with the help of probiotic microorganisms.

Highlighting the Importance of Matrix Metalloproteinase 1, 8, and 9 Expression during the Progression of Mycobacterium tuberculosis Infection

Sasikumar Pitchaikani — 2024-03-15

Tuberculosis (TB) is one of the major threats to public health; annually it kills more than 1.5 million people around the globe. Tuberculosis is caused by an intracellular pathogen named Mycobacterium tuberculosis (Mtb). This Mtb enters the lung through the respiratory passage by inhalation in healthy individuals. Infection of this disease starts from the settlement of Mtb to the lung alveoli of the host from the external bacilli air droplets. After settlement, the multiplication of Mtb results in the induction of innate immunity through the alveolar macrophages. Compared to other infectious diseases, tuberculosis infection was transmitted rapidly by the infected aerosols released from infected persons to healthy persons through the air. After infection, disease development results in the formation of drug-resistance TB (DR-TB) with four subcategories, i.e. Single-drug resistant TB (SDR-TB), multi-drug resistant TB (MDR-TB), extensive drug-resistant TB (XDR-TB), and total-drug resistant TB (TDR-TB). As a result, this DR-TB may act as a major source of TB death due to spontaneous antimicrobial resistance (AMR). This AMR makes the anti-TB drugs ineffective. In the current scenario, researchers are trying to find the drug target to decrease tuberculosis progression instead of drug resistance. The present review reports that the outcome of research studies showed that matrix metalloproteinase (MMP) may act as a suitable target for treating Mtb infection with the help of specific proteinase inhibitors. Recent reports have shown the specific role of matrix metalloproteinases 1, 8, and 9 in the disease progression and its role in normal homeostasis mechanism with the help of specific animal models/In vitro models.

Exploration and Profiling of Potential Thermo-alkaliphilic Bacillus licheniformis and Burkholderia sp. from varied Soil of Delhi region, India and their Plant Growth-Promoting Traits

Charu Singh — 2024-03-15

Soilless cultivation has emerged as a fundamental alternative for large-scale vegetable production because it generates high-quality yields and uses resources efficiently. While plant growth-promoting bacteria (PGPB) are known to enhance growth and physiological aspects in crops grown in soil, their application in soilless cultivation has been relatively less explored. This study aimed to isolate potential PGPBs from soil samples collected from five locations in and around the Delhi-National Capital Region (NCR), India, which were further screened for significant PGPB attributes. Among these, 51 isolated were selected for assessing the impact on Oryza sativa (rice) growth and yield grown on a hydroponic set. The results indicated that isolates AFSI16 and ACSI02 significantly improved the physiological parameters of the plants. For instance, treatment with AFSI16 showed a 23.27% increase in maximum fresh shoot mass, while ACSI02 resulted in a 46.8% increase in root fresh mass. Additionally, ACSI02 exhibited the highest shoot length (34.07%), whereas AFSI16 exhibited the longest root length (46.08%) in O.sativa. Treatment with AFSI16 also led to significant increases in total protein content (4.94%) and chlorophyll content (23.44%), while ACSI02 treatment showed a 13.48% increase in maximum carotenoid content in the leaves. The potential PGPBs were identified through 16S rRNA sequencing, as the two most effective strains, AFSI16 and ACSI02, belonged to thermo-alkaliphilic Bacillus licheniformis and Burkholderia sp., respectively. This study demonstrated the potential of these identified PGPB strains in enhancing crop performance, specifically in soilless cultivation systems.

Phytotoxicity and genotoxicity assessment of organic and inorganic contaminants detected in pharmaceutical industrial wastewaters using Vigna radiata and Allium cepa

Km Jyoti — 2024-03-15

The discharged effluent of pharmaceutical industrial wastewater treatment plants (PIWWTPs) exhibits substantial environmental toxicity due to the intricate combination of organic and inorganic pollutants. This study assessed the phytotoxicity, genotoxicity, and cytotoxicity of untreated and treated pharmaceutical industrial wastewater (PIWW). Most of the physicochemical parameters viz. COD, BOD, EC, sulfide, sulfate, nitrate, phosphate, grease, phenols, and metal concentrations viz. B, Cr, Ca, Cd, Cu, Zn, Pb, Hg, and As in untreated wastewater (UTW) were noted beyond the permissible limit and remained higher in treated wastewater (TW). The findings revealed that the performance of PIWWTP was woefully inadequate. The GC-MS spectra of UTW and TW revealed the presence of various organic contaminants. The toxicological studies showed that the UTW had a high degree of phytotoxicity, which persisted even after the treatment as it inhibited the seed germination in Vigna radiata. The seed germination was inhibited up to 70% and 50% tested at 50% concentration of UTW and TW respectively. Genotoxicity was measured by determining mitotic index and chromosomal aberrations in Allium cepa root apex grown in untreated and treated PIWW. Compared to the negative control, the mitotic index dropped to 85% and 75% at the 50% concentrations of UTW and TW, respectively. Chromosomal aberrations were also found in the cellular mass of root apex growing in both UTW and TW. According to the findings, it is unsafe for the environment to release PIWW that has not been properly treated, as this could pose serious risks to environmental health.

Ethnopharmacological study of medicinal plants used in the treatment of skin diseases in the Western Middle Atlas region (Morocco)

Fatiha El Azzouzi — 2024-03-15

An investigation was conducted among 360 people from the local population of the Western Middle Atlas of Morocco to identify medicinal plants used for treating skin diseases. Various parameters, including Relative Frequency of Citation (RFC), Family Use Value (FUV), Plant Part Value (PPV), Informant Consensus Factor (ICF), and Fidelity Level (FL), were used for data collection and assessment. During the investigation, 45 medicinal plant species belonging to 33 families were documented, with the most important family being Euphorbiaceae (FUV = 0.292). The highest ICF value (ICF=0.991) was mentioned for skin cancer. The poultice was found to be the primary method for preparing the majority of remedies (51%). Leaves were the most commonly used plant part (PPV = 0.476), and Allium sativum L. was the most widely used species (RFC = 0.302). These findings are a preliminary step towards conserving and popularising these plant species, promoting sustainable practices in traditional medicine, safeguarding biodiversity, and integrating these valuable botanical resources into modern healthcare systems.

Effects of Elicitation on Invitro Regeneration of two Tomato (Solanum lycopersicum L.) Cultivars in Tissue Culture

Alhagie K. Cham — 2024-03-15

Exploring alternative avenues, in vitro culture emerges as a promising option for potential bioactive compound sources. However, compared to intact plants, only a few cultures demonstrate efficient synthesis of secondary metabolites. Elicitors have gained prominence as stress agents for enhancing in vitro micropropagation in specific tissues, organs, and cells. Recent advancements in plant tissue culture involve elicitors, opening new possibilities for in vitro production of crucial food crops. This research aimed to investigate the impact of three elicitors (Activane®, Micobiol®, and Stemicol®) on germination and in vitro multiplication of two tomato cultivars explants, employing both direct and indirect in vitro organogenesis. Among the texted elicitors, Micobiol® emerged as a successful elicitor, promoting optimal seed germination, survival, and 100% growth compared to the 80% in the control group. Further, Activane® exhibited a favourable induction response and achieved 96%, 95%, and 100% in weight and diameter of callus, yet various elicitor concentrations did not exert significant influence across treatments. In conclusion, an effective disinfection and in vitro implantation of tomato seeds ensured successful germination, promoting seedling survival and growth. Various elicitors positively impacted in vitro organogenesis, particularly in root induction, with higher survival percentages in acclimatized plants. The study guides future research on elicitor treatments for large-scale tomato in vitro propagation, emphasizing the need to identify optimal elicitor concentrations.

Analyzing the antimicrobial efficacy of the economically important tree Knema linifolia (Roxb.) Warb

Ria Bhar — 2024-03-15

Knema linifolia is widely used for fuel wood, fodder and healthcare purposes. This plant treats various diseases in different parts of India, including Assam, Meghalaya, Alipurduar and Darjeeling districts of West Bengal. This study was carried out to determine the bactericidal properties of various parts of K. linifolia aqueous extract. The aqueous extract of the leaves, bark, stem and plant sap were tested against Escherichia coli (gram-negative bacteria) & Staphylococcus aureus (gram-positive bacteria). Among the tested extracts, both the leaf and bark extracts were found to have high bactericidal potential and can kill more than 60% of both bacterial strains with a concentration of 300µg/mL through an agar diffusion test. The MIC (Minimum Inhibitory Concentration) values for the leaf and bark extracts were recorded at ≤1000µg/mL & ≤500µg/mL, respectively. It has also been found that both the bark and leaf extracts contain high tannins, which might be essential for the antibacterial properties of Knema sp. There is currently a lack of proper documentation on using K. linifolia, which makes it challenging to conduct clinical or commercial research to support new uses in modern phototherapy. This study aims to fill this gap and provide significant information that could lead to changes in modern medicine.

Molecular regulation of Mycobacterium tuberculosis Sigma factor H with Anti-sigma factor RshA under stress condition

Sayantee Sen — 2024-03-15

Mycobacterium tuberculosis is the causative agent of tuberculosis, the leading fatal infectious disease that claims millions of lives every year. M. tuberculosis regulates its stress condition response using its regulatory protein, Sigma Factor H, which binds with its cognate anti-sigma factor RshA in normal conditions, forming a complex inhibiting transcription. During oxidative stress, SigH is released from the complex and binds to RNA Polymerase (RNAP) to initiate transcription. Thus, it is important to understand the molecular conformational state of SigH in complex with different protein partners under different cellular or environmental contexts. This work intends to analyze the SigH-RshA complex, which revealed the variation in SigH shown during complex formation with RNAP and RshA, respectively. Previously, Hydrogen Deuterium Exchange-Mass Spectrometry (HDX-MS) analysis of SigH-RshA interaction provided a detailed insight into the critical residues participating in the interaction. The HDX-MS data were used to dock RshA on the open conformation of SigH from the SigH-RNAP complex structure (PDB: 5ZX2), and closed conformation was obtained from protein modelling. The docking revealed that closed conformation of SigH complexing with RshA in terms of HDX-MS data revealed a major structural shift in SigH while interacting with two different binding partners, RshA and RNAP, under variable environmental conditions. This structural shift of SigH with RshA and RNAP has significance in understanding the stress response of M. tuberculosis, and SigH could prove to be a potential drug target.

ARTIFICIAL INTELLIGENCE IN TACKLING CORONAVIRUS AND FUTURE PANDEMICS

Shagufta Quazi — 2024-03-15

SARS-COV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) was initially tested in Wuhan City, China, in December 2019 and had a devastating impact worldwide, exterminating more than 6 million people as of September 2022. It became the biggest worldwide health crisis since the 1918 influenza outbreak. Viruses generally mutate randomly, so predicting how SARS-CoV-2 will transform over the next few months or years and which forms will predominate is impossible. The possibilities for virus mutation, in theory, are practically endless. Enabling researchers to determine which antibodies have the potential to be most effective against existing and future variations could help machine learning to assist in drug discovery. In the COVID-19 pandemic, AI has benefited four key areas: diagnosis, clinical decision-making for public health, virtual assistance, and therapeutic research. This study conducted a discourse analysis and textual evaluation of AI (deep learning and machine learning) concerning the COVID-19 outbreak. Further, this study also discusses the latest inventions that can be very helpful in future pandemic detection. COVID-19 has already changed our lives, and in the future, we might be able to deal with pandemics like this with the help of AI. This review has also emphasized the legal implications of AI in the battle against COVID-19.

Residue-specific orientation of arrestin in 5-HTR1B (Serotonin Receptor)- βArrestin-1 interaction

Somdatta Bhattacharya — 2024-03-15

Physiologically G protein-coupled receptors (GPCRs) are an important class of cell surface proteins capable of sensing the exogenous signals across the cell membrane through G-protein-dependent and independent pathways. Activated GPCRs initiate diverse G-protein-independent signalling through interaction with arrestin. Arrestins comprise a family of four proteins that act as signal regulators of GPCRs. Arrestin specificity and assembly orientation with a particular GPCR depend on the finger loop's residues. Recent cryo-EM structural elucidation of neurotensin receptor-1(NTSR1)-β-arrestin1complex reveals its striking difference from Rhodopsin-visual-Arrestin by a 90˚ rotation of β-Arrestin1 concerning the receptor. Alignment of neurotensin receptor 1(NTSR1)-β-Arrestin1 assembly with 5-HTR_1B (Serotonin receptor) structure shows an ionic interaction mediated complex formation between receptor binding cleft and finger loop of arrestin. Mutational analysis of finger loop residues R65, D67, and D69 of β-Arrestin1 by tango assay confirms its possible interaction with an electropositive pocket of K79 and R161 in 5-HTR_1B.