Submerged cultivation and phytochemical analysis of medicinal mushrooms (Trametes sp.).

Mushrooms are widely available around the world and have various nutritional as well as therapeutic values. Many Asian cultures believe that medicinal mushrooms can prolong life and improve vitality. This study aims to characterize the phytochemical and polysaccharide content, mainly ß-glucan content, of mycelial biomass and fruiting bodies collected from the Himalayan region, particularly Uttarakhand. Through molecular analysis of the LSU F/R-rDNA fragment sequence and phylogenetic analysis, the strain was identified as Trametes sp. We performed screening of phytochemicals and polysaccharides in mushroom and biomass extracts using high-performance liquid chromatography (HPLC) and a PC-based UV-Vis spectrophotometer. The macrofungal biomass was found to be high in saponin, anthraquinone, total phenolic, flavonoid, and ß-glucan content. In biomass extract, we observed a high level of saponin (70.6µg/mL), anthraquinone (14.5µg/mL), total phenolic (12.45 µg/mL), and flavonoid (9.500 µg/mL) content. Furthermore, we examined the contents of alkaloids, tannins, terpenoids, and sterols in the biomass and mushroom extracts; the concentration of these compounds in the ethanol extract tested was minimal. We also looked for antioxidant activity, which is determined in terms of the IC50 value. Trametes sp. mushroom extract exhibits higher DPPH radical scavenging activity (62.9% at 0.5 mg/mL) than biomass extract (59.19% at 0.5 mg/mL). We also analyzed ß-glucan in Trametes sp. from both mushroom and biomass extracts. The biomass extract showed a higher ß-glucan content of 1.713 mg/mL than the mushroom extract, which is 1.671 mg/mL. Furthermore, ß-glucan analysis was confirmed by the Megazyme ß-glucan assay kit from both biomass and mushroom extract of Trametes sp. ß-glucans have a promising future in cancer treatment as adjuncts to conventional medicines. Producing pure ß-glucans for the market is challenging because 90-95% of ß glucan sold nowadays is thought to be manipulated or counterfeit. The present study supports the recommendation of Trametes sp. as rich in ß-glucan, protein, phytochemicals, and antioxidant activities that help individuals with cancer, diabetes, obesity, etc.

Development of lead (Pb) tolerant strain by protoplast technology and their remediation.

The lead is poisonous metal and because of its chemical nature it acts as an environmental contaminant through the water or soil and it becomes toxic to humans. The toxicity of Pb occurs as a change in the conformation of nucleic acid and protein, inhibition of enzyme activity, disruption of membrane function and oxidative phosphorylation. For protoplast preparation, the removal of the cell wall and protoplast formation obtained by specific lytic enzyme. In cytoplasmic membrane, the envelope of bacteria consists of overlying cell wall. From hypertonic environment, the complete cell wall removal occurs due to which it maintains the osmotic integrity of the cell and produces the protoplast. In current work, protoplasts were produced by specific lytic enzyme (lysozyme and macerozyme), chemo fused (with the help of Polyethylene Glycol) and regenerated from strains Staphylococcus sp. and Bacillus sp. The fused protoplast was spherical in shape observed under microscopy. Colonies were screened on specific medium supplemented with Pb (Concentration at the rate of 1.5mM). One resistant colony (MICBT-1) was selected and further examined and applied for the transformation of Pb in the broth medium. The strain removed 98% of Pb at 1mM concentration. Next, sucrose containing medium was best which gives maximum protoplast regeneration. From various organisms, fusion technique has been used to combine the genes to create the strains having desired properties. This is a significant technique for engineering of bacterial strains for advantageous applied properties. Further MICBT-1 applied in artificially contaminated soil and removed maximally in exchangeable fraction (remains up to 0.05 mM). An efficient bioremediating agent for lead transformation from soil and water is expected to ease the ever-increasing problem. Further, it is needful to obtain new strain with the help of protoplast technology which can reduce the pollutant. This lead tolerant strain can be applied for bioremediation purposes in the Pb contaminated soil and water environment.

Mushrooms as Potential Sources of Active Metabolites and Medicines.

Background: Mushrooms exist as an integral and vital component of the ecosystem and are very precious fungi. Mushrooms have been traditionally used in herbal medicines for many centuries. Scope and Approach: There are a variety of medicinal mushrooms mentioned in the current work such as Agaricus, Amanita, Calocybe, Cantharellus, Cordyceps, Coprinus, Cortinarius, Ganoderma, Grifola, Huitlacoche, Hydnum, Lentinus, Morchella, Pleurotus, Rigidoporus, Tremella, Trametes sp., etc., which play a vital role in various diseases because of several metabolic components and nutritional values. Medicinal mushrooms can be identified morphologically on the basis of their size, color (white, black, yellow, brown, cream, pink and purple-brown, etc.), chemical reactions, consistency of the stalk and cap, mode of attachment of the gills to the stalk, and spore color and mass, and further identified at a molecular level by Internal Transcribed Spacer (ITS) regions of gene sequencing. There are also other methods that have recently begun to be used for the identification of mushrooms such as high-pressure liquid chromatography (HPLC), nuclear magnetic resonance spectroscopy (NMR), microscopy, thin-layer chromatography (TLC), DNA sequencing, gas chromatography-mass spectrometry (GC-MS), chemical finger printing, ultra-performance liquid chromatography (UPLC), fourier transform infrared spectroscopy (FTIR), liquid chromatography quadrupole time-of-flight mass spectrometry (LCMS-TOF) and high-performance thin-layer chromatography (HPTLC). Lately, the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) technique is also used for the identification of fungi. Key Finding and Conclusion: Medicinal mushrooms possess various biological activities like anti-oxidant, anti-cancer, anti-inflammatory, anti-aging, anti-tumor, anti-viral, anti-parasitic, anti-microbial, hepatoprotective, anti-HIV, anti-diabetic, and many others that will be mentioned in this article. This manuscript will provide future direction, action mechanisms, applications, and the recent collective information of medicinal mushrooms. In addition to many unknown metabolites and patented active metabolites are also included.

Diagnosis, prevention, and treatment of coronavirus disease: a review.

INTRODUCTION: Coronavirus disease (COVID-19) was first reported in Wuhan, China, in late December 2019 and subsequently, declared a pandemic. As of 3 June 2021, 172,493,290 individuals have acquired COVID-19 and 3,708,334 patients have died worldwide, according to the World Health Organization. AREAS COVERED: This review explores epidemiology; virology; pathogenesis; genomic variations; mode of transmission; clinical occurrence; diagnosis; and treatment with antiviral agents, antibiotics, and supportive therapies. It covers a nanotechnology-based treatment approach and emphasizes the importance of herbal and marine antiviral drugs. The review attempts to explain current advances in research, prevention, and control of COVID-19 spread through artificial intelligence and vaccine development status under cosmopolitan consideration. EXPERT OPINION: While COVID-19 research is advancing at full capacity, the discovery of drugs or vaccines that can fight the pandemic is necessary. Human survival in such a critical situation will be possible only with the development of strong immunity by opting for exercise, yoga, and consumption of hygienic food and beverages. Therefore, education about COVID-19 lethality and its impact on livelihood is important. The pandemic has also shown positive effects on the environment, such as a significant reduction in environmental pollution and global warming and improvement in river water quality.

In-situ remediation of nitrogen and phosphorus of beverage industry by potential strains Bacillus sp. (BK1) and Aspergillus sp. (BK2).

The bioremediation of beverage (treated and untreated) effluent was investigated in the current study by using the potential strains of Bacillus sp. (BK1) and Aspergillus sp. (BK2). Effluent was collected from the beverage industry (initial concentration of nitrogen were 3200 ± 0.5 mg/L and 4400 ± 0.6 mg/L whereas phosphorus were 4400 ± 2 mg/L and 2600 ± 1 mg/L in treated and untreated effluent correspondingly). Further, the BK1 and BK2 exhibited high removal competence after 1 week of incubation; BK1 removed phosphorus 99.95 ± 0.7% and BK2 95.69 ± 1% in treated effluent while nitrogen removed about 99.90 ± 0.4% by BK1 and 81.25 ± 0.8% by BK2 (initial concentration of phosphorus 4400 ± 2 mg/L and nitrogen 3200 ± 0.5 mg/L). Next, in the untreated effluent BK1 removed 99.81 ± 1% and BK2 99.85 ± 0.8% of phosphorus while removed nitrogen 99.93 ± 0.5% by BK1 and 99.95 ± 1.2% by BK2 correspondingly, (initial concentration of phosphorus 2600 ± 1 mg/L and nitrogen 4400 ± 0.6 mg/L). The physiochemical composition of sample such as pH, total carbohydrates, total proteins, total solids of treated and untreated effluent were also analysed before and after treatment of both the samples. BK1 and BK2 increased the pH by 8.94 ± 0.3 and 9.5 ± 0.4 correspondingly in treated effluent whereas 6.34 ± 0.5 and 7.5 ± 0.2 correspondingly in untreated effluent (initial pH of treated and untreated effluent 7.07 ± 0.8 and 4.85 ± 0.3 correspondingly). Total Carbohydrates removed about 17,440 ± 4.6 mg/L and 10,680 ± 3.2 mg/L by BK1 and BK2 correspondingly in treated effluent whereas 18,050 ± 3.5 mg/L and 18,340 ± 2.3 mg/L correspondingly in untreated effluent (initial concentration of treated and untreated effluent 25,780 ± 1.6 mg/L and 35,000 ± 1.5 mg/L correspondingly) while BK1 and BK2 removed total proteins by 30.336 ± 4.6 mg/L and 40.417 ± 2.3 mg/L correspondingly in treated effluent whereas 18.929 ± 1.2 mg/L and 17.526 ± 0.8 mg/L correspondingly in untreated effluent (initial concentration of treated and untreated effluent 49.225 ± 1.5 mg/L and 20.565 ± 1 mg/L correspondingly). Next, total solids removed by BK1 and BK2 2.5 ± 0.3 mg/L and 1.6 ± 0.6 mg/L correspondingly in treated effluent whereas 5.5 ± 0.8 mg/L and 4.6 ± 0.6 mg/L in untreated effluent (initial concentration of treated and untreated effluent 5.6 ± 1.5 mg/L and 9.48 ± 1.2 mg/L correspondingly). Both the strains BK1 and BK2 are highly efficient in the nitrogen and phosphorus removal therefore this strain may be applied for the potential remediation.

Microbial Fabricated Nanosystems: Applications in Drug Delivery and Targeting.

The emergence of nanosystems for different biomedical and drug delivery applications has drawn the attention of researchers worldwide. The likeness of microorganisms including bacteria, yeast, algae, fungi, and even viruses toward metals is well-known. Higher tolerance to toxic metals has opened up new avenues of designing microbial fabricated nanomaterials. Their synthesis, characterization and applications in bioremediation, biomineralization, and as a chelating agent has been well-documented and reviewed. Further, these materials, due to their ability to get functionalized, can also be used as theranostics i.e., both therapeutic as well as diagnostic agents in a single unit. Current article attempts to focus particularly on the application of such microbially derived nanoformulations as a drug delivery and targeting agent. Besides metal-based nanoparticles, there is enough evidence wherein nanoparticles have been formulated using only the organic component of microorganisms. Enzymes, peptides, polysaccharides, polyhydroxyalkanoate (PHA), poly-(amino acids) are amongst the most used biomolecules for guiding crystal growth and as a capping/reducing agent in the fabrication of nanoparticles. This has promulgated the idea of complete green chemistry biosynthesis of nano-organics that are most sought after in terms of their biocompatibility and bioavailability.

Insights of Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) pandemic: a current review.

COVID-19, a pandemic of the 21st century caused by novel coronavirus SARS-CoV-2 was originated from China and shallowed world economy and human resource. The medical cures via herbal treatments, antiviral drugs, and vaccines still in progress, and studying rigorously. SARS-CoV-2 is more virulent than its ancestors due to evolution in the spike protein(s), mediates viral attachment to the host's membranes. The SARS-CoV-2 receptor-binding spike domain associates itself with human angiotensin-converting enzyme 2 (ACE-2) receptors. It causes respiratory ailments with irregularities in the hepatic, nervous, and gastrointestinal systems, as reported in humans suffering from COVID-19 and reviewed in the present article. There are several approaches, have been put forward by many countries under the world health organization (WHO) recommendations and some trial drugs were introduced for possible treatment of COVID-19, such as Lopinavir or Ritonavir, Arbidol, Chloroquine (CQ), Hydroxychloroquine (HCQ) and most important Remdesivir including other like Tocilizumab, Oritavancin, Chlorpromazine, Azithromycin, Baricitinib, etc. RT-PCR is the only and early detection test available besides the rapid test kit (serodiagnosis) used by a few countries due to unreasonable causes. Development of vaccine by several leader of pharmaceutical groups still under trial or waiting for approval for mass inoculation. Management strategies have been evolved by the recommendations of WHO, specifically important to control COVID-19 situations, in the pandemic era. This review will provide a comprehensive collection of studies to support future research and enhancement in our wisdom to combat COVID-19 pandemic and to serve humanity.

Sludge: next paradigm for enzyme extraction and energy generation.

Excess of sludge generated during activated sludge process by various industries is causing sludge disposable problem worldwide. Sludge contains organic materials of a highly complex mixture of molecules with some toxic compounds. Thus, the harmful effect of sludge could be managed by processing of sludge and recovery of useful products. Sludge could undergo biodegradation and used for the recovery of enzymes and energy. Enzymes (amylase, alkaline phosphatase, lipase, proteases) are produced by microorganisms and released into the media to degrade organic matters of sludge. These enzymes can be extracted by means of different physical and chemical methods from activated sludge for the multifarious application. Current manuscript discussed all the methods utilized for the extraction of enzyme and their utilization in terms of energy generation. Extracted industrial enzymes are used in agriculture, dairy, detergents, pulp, paper, cosmetics, and pharmaceutics. Apart from enzyme extraction, production of energy (biofuels, electricity) is also done by key treatment of sludge. It is a compilation of current knowledge and expected competitiveness with respect to existing methods already applied in practice for enzyme recovery. This paper presents an overview of the production of valuable enzymes and various forms of energy from sludge, which is toxic and unwanted for life.

Bio-transformation and stabilization of arsenic (As) in contaminated soil using arsenic oxidizing bacteria and FeCl3 amendment.

A combination of biological and chemical methods was applied in the present study to evaluate the removal of arsenic (As) from contaminated soil. The treatment involved As-oxidizing microbes aimed of transforming the more toxic As (III) to less toxic As (V) in the soil. FeCl3 was added at three different concentrations (1, 2, and 3%) to stabilize the As (V). Leaching of the treated soil was investigated by making a soil column and passing tap water through it to determine solubility. Experimental results indicated that the bacterial activity had a pronounced positive effect on the transformation of As, and decreased the soluble exchangeable fraction from 50 to 0.7 mg/kg as compared to control and from 50 to 44 mg/kg after 7 days of treatment. FeCl3 also played an indispensable role in the adsorption/stabilization of As in the soil; 1 and 2% FeCl3 strongly influenced the adsorption of As (V). The soil leachate contained negligible amount of As and trace metals, which indicates that combining an efficient microbe with a chemical treatment is very effective route for the removal and stabilization of As from contaminated soil in the environment.

Pseudomonas sp. CL7 from Sludge Removed 2,3,4,6-Tetrachlorophenol in Vivo and in Vitro Condition.

The present research focused on 2,3,4,6-Tetrachlorophenol (2,3,4,6-TeCP) mineralizing bacterium from the sludge of pulp and paper industry and identified as Pseudomonas sp. CL7 by 16s rRNA gene sequences analysis. This isolate degraded 2,3,4,6-TeCP as indicated by stoichiometric release of chloride and biomass formation. High pressure liquid chromatography (HPLC) analysis showed that Pseudomonas sp. (CL7) was able to mineralize a higher concentration of 2,3,4,6-TeCP (600 mg/l or 2.5 mM) than any previously reported 2,3,4,6-TeCP degrading bacteria. As the concentration of 2,3,4,6-TeCP increased from 50 (0.21 mM) to 600 mg/l (2.5 mM), the reduction in the cell growth was observed and the 2,3,4,6-TeCP degradation was more than 85% in all the concentrations in the present study. CL7 was able to remove 100% of 2,3,4,6-TeCP from the sludge (in Vitro condition) when supplemented with 100 mg/l (0.42 mM) of 2,3,4,6-TeCP and grown for two weeks. This study showed that CL7 can be used for bioremediation of 2,3,4,6-TeCP.

Arsenic (As) contamination: A major risk factor in Xinjiang Uyghur autonomous region of China.

Xinjiang province is one of the unhealthiest region in China due to natural as well as man-made activities. Here soil and water having high concentration of multi-metals especially arsenic content in the soil and water is a major threat to the peoples which suffers regularly from arsenic contamination therefore multiple diseases and illness is the common phenomenon. Therefore this area urgently needed a comprehensive assessment by governmental and nongovernmental organization to cope up with this problem and find a miracle solution which can remediate soil and water quality. There peoples suffers much for the above mentioned reason.

Pentachlorophenol remediation by Enterobacter sp. SG1 isolated from industrial dump site.

Chlorophenols contamination is serious concern to the environment due toxicity to all forms of life. Among all the chlorophenols, pentachlorophenol (PCP) is more detrimental to the environment. Pentachlorophenol used as pesticide, herbicide, antifungal agent and wood preservative which causes environmental pollution. In the present research a PCP degrading bacterium was isolated and characterized from industrial dump site. This isolate used PCP as its sole source of carbon and energy and was capable of degrading this compound, as indicated by stoichiometric release of chloride, ring cleavage activity and biomass formation. Based on morphological, biochemical and 16S rRNA gene sequence analysis this strain was identified as Enterobacter sp. SG1. Gas Chromatography (GC) analysis revealed that this strain was able to degrade PCP up to a concentration of 2 mM. This study showed that the removal efficiency of PCP by SG1 was found to be very effective and can be used in degradation of PCP contaminated site or waste in the environment.

Cross-sectional study of Trichinella spp in pigs in CDR, Nepal using pepsin digestion and ELISA serology.

This epidemiological cross-sectional study was conducted in five major pig-producing districts of CDR, Nepal from November 2006 to April 2007. A total of 576 slaughtered pigs were randomly selected and diaphragmatic crus muscles (n = 551), corresponding sera (n = 487) and sera only (n = 25) were collected from them. Meat samples were examined by pepsin digestion to evaluate for larvae of Trichinella spp. The sera were investigated using ELISA to evaluate for antibodies against T. spiralis. The doubtful and positive sera from the ELISA test were investigated by end-point ELISA, and the Western blot was used for confirmatory diagnosis. Pepsin digestion did not detect the larvae of Trichinella spp. AB-ELISA showed 2 positive and 14 doubtful results, out of 344 sera analyzed. End-point ELISA and the Western blot had revealed that all 16 samples were truly negative. A questionnaire revealed no rodent control (70%), left over feeding practices (65%), dumping of farm waste (82.5%) and uncooked meat was being used as feed (100%). This study reveals that Trichinella spp has a low prevalence.