In vitro and in silico investigation of effects of antimicrobial peptides from Solanaceae plants against rice sheath blight pathogen Rhizoctinia solani.

Rhizoctonia solani, the causative agent of sheath blight disease in rice, poses a significant threat to agricultural productivity. Traditional management approaches involving chemical fungicides have been effective but come with detrimental consequences for the ecosystem. This study aimed to investigate sustainable alternatives in the form of antifungal peptides derived from Solanaceous plant species as potential agents against R. solani. Peptide extracts were obtained using an optimized antimicrobial peptide (AMP) extraction method and desalted using the solid-phase extraction technique. The antifungal potential of peptide-rich extracts from Solanum tuberosum and Capsicum annum was assessed through in vitro tests employing the agar well diffusion method. Furthermore, peptide-protein docking analysis was performed on HPEPDOCK and HDOCK server; and molecular dynamics simulations (MDS) of 100 ns period were performed using the Gromacs 2020.4. The results demonstrated significant inhibition zones for both extracts at concentrations of 100 mg/mL. Additionally, the extracts of Solanum tuberosum and Capsicum annum had minimum inhibitory concentrations of 50 mg/mL and 25 mg/mL, respectively with minimum fungicidal concentrations of 25 mg/mL. Insights into the potential mechanisms of key peptides inhibiting R. solani targets were gleaned from in-silico studies. Notably, certain AMPs exhibited favorable free energy of binding against pathogenicity-related targets, including histone demethylase, sortin nexin, and squalene synthase, in protein-peptide docking simulations. Extended molecular dynamics simulations lasting 100 ns and MM-PBSA calculations were performed on select protein-peptide complexes. AMP10 displayed the most favorable binding free energy against all target proteins, with AMP3, AMP12b, AMP6, and AMP15 also exhibiting promising results against specific targets of R. solani. These findings underscore the potential of peptide extracts from S. tuberosum and C. annum as effective antifungal agents against rice sheath blight caused by R. solani.

Bio-Fabrication of Silver Nanoparticles Using Citrus aurantifolia Fruit Peel Extract (CAFPE) and the Role of Plant Extract in the Synthesis.

The green synthesis of silver nanoparticles has been proposed as an eco-friendly and cost-effective substitute for chemical and physical methods. The aim of this study was to synthesize and characterize silver nanoparticles using the peel extract of Citrus aurantifolia fruit, and to determine the possible phytochemical constituents' presence in the plant extracts that might be responsible for the synthesis. Citrus aurantifolia fruit peel extraction was followed by phytochemical studies of secondary metabolites, FTIR analysis confirmation of functional groups, and GC-MS analysis. Silver nanoparticles were synthesized through bio-reduction of silver ions (Ag+) to silver nanoparticles using CAFPE and characterized using UV-Vis spectroscopy, HR-TEM, FESEM, EDX, XRD, DLS, and FTIR. The presence of plant secondary metabolites such as alkaloids, flavonoids, tannins, saponins, phenols, terpenoids, and steroids was detected. The FTIR analysis of the extract revealed the presence of functional groups like hydroxyl, carboxyl, carbonyl, amine, and phenyl, whereas the GC-MS analysis indicated presence of chemical compounds such as 1,2,4-Benzenetricarboxylic acid, Fumaric acid, nonyl pentadecyl, and 4-Methyl-2-trimethylsilyloxy-acetophenone, etc., with similar functional groups. The synthesized silver nanoparticle (AgNP) has displayed the characteristics of a surface plasmon resonance (SPR) band peak from 360-405 nm. High resolution transmission electron microscope (HR-TEM) and field emission scan electron microscope (FESEM) confirm polydisperse, spherical shaped, and smooth surface nanoparticles with an average size of 24.023 nm. Energy dispersive X-ray (EDX) analysis further revealed that silver is the most abundant element found in the micrograph of the nanoparticles, and FTIR analysis further confirmed the presence of different functional groups in the surface of the nanoparticle. The XRD analysis also confirmed that the nanoparticles synthesized are crystalline in nature. Based on the findings of this study, it is understood that the variety of natural compounds that are present in plant extracts of Citrus aurantifolia fruit peel can act as both reducing and stabilizing agents for the synthesis of silver nanoparticles. It is, therefore, concluded that Citrus aurantifolia peel extract can be potentially used for the large production of silver nanoparticles for several applications.

A Review on Plants and Microorganisms Mediated Synthesis of Silver Nanoparticles, Role of Plants Metabolites and Applications.

Silver nanoparticles are one of the most extensively studied nanomaterials due to their high stability and low chemical reactivity in comparison to other metals. They are commonly synthesized using toxic chemical reducing agents which reduce metal ions into uncharged nanoparticles. However, in the last few decades, several efforts were made to develop green synthesis methods to avoid the use of hazardous materials. The natural biomolecules found in plants such as proteins/enzymes, amino acids, polysaccharides, alkaloids, alcoholic compounds, and vitamins are responsible for the formation of silver nanoparticles. The green synthesis of silver nanoparticles is an eco-friendly approach, which should be further explored for the potential of different plants to synthesize nanoparticles. In the present review we describe the green synthesis of nanoparticles using plants, bacteria, and fungi and the role of plant metabolites in the synthesis process. Moreover, the present review also describes some applications of silver nanoparticles in different aspects such as antimicrobial, biomedicine, mosquito control, environment and wastewater treatment, agricultural, food safety, and food packaging.

A Review on the Prevalence of Toxoplasma gondii in Humans and Animals Reported in Malaysia from 2008-2018.

Toxoplasmosis is a disease caused by the protozoan parasite Toxoplasma gondii (T. gondii). Human toxoplasmosis seroprevalence in Malaysia has increased since it was first reported in 1973 as shown in previous reviews of 1991 and 2007. However, over a decade since the last review, comprehensive data on toxoplasmosis in Malaysia is lacking. This work aimed at reviewing articles on toxoplasmosis research in Malaysia in order to identify the research gaps, create public awareness, and efforts made so far and proffer management options on the disease. The present review examines the available published research articles from 2008 to 2018 related to toxoplasmosis research conducted in Malaysia. The articles reviewed were retrieved from nine credible databases such as Web of Science, Google Scholar, ScienceDirect, PubMed, Scopus, Springer, Wiley online library, Ovid, and Cochrane using the keywords; Malaysia, toxoplasmosis, Toxoplasma gondii, toxoplasma encephalitis, seroprevalence, human immunodeficiency virus (HIV) patients, pregnant women, genotype strain, anti-toxoplasma antibodies, felines, and vaccine. The data highlighted seropositive cases from healthy community members in Pangkor Island (59.7%) and among migrant workers (57.4%) at alarming rates, as well as 42.5% in pregnant women. Data on animal seroprevalence were limited and there was no information on cats as the definitive host. Genetic characterization of Toxoplasma gondii from HIV patients; pregnant women, and domestic cats is lacking. This present review on toxoplasmosis is beneficial to researchers, health workers, animal health professionals, and policymakers. Therefore, attention is required to educate and enlighten health workers and the general public about the risk factors associated with T. gondii infection in Malaysia.

Phytochemicals and Potential Therapeutic Targets on Toxoplasma gondii Parasite.

Identification of drug target in protozoan T. gondii is an important step in the development of chemotherapeutic agents. Likewise, exploring phytochemical compounds effective against the parasite can lead to the development of new drug agent that can be useful for prophylaxis and treatment of toxoplasmosis. In this review, we searched for the relevant literature on the herbs that were tested against T. gondii either in vitro or in vivo, as well as different phytochemicals and their potential activities on T. gondii. Potential activities of major phytochemicals, such as alkaloid, flavonoid, terpenoids and tannins on various target sites on T. gondii as well as other related parasites was discussed. It is believed that the phytochemicals from natural sources are potential drug candidates for the treatment of toxoplasmosis with little or no toxicity to humans.

Susceptibility of Toxoplasma gondii to Ethanolic Extract of Tinospora crispa in Vero Cells.

BACKGROUND: Toxoplasmosis remains widely distributed globally and is one of the major neglected parasitic zoonotic infections. The infection is still endemic in most parts of the world due to poor control as well as challenges of the currently used medications which can be overcome by using natural products. This study evaluated the effect of ethanolic extract from the stem of Tinospora crispa (EETC) on host cell invasion and intracellular replication of Toxoplasma gondii. METHOD: The stem powder of T. crispa was soaked in absolute ethanol for 72 hours. The resulting ethanolic extract was screened for the presence of phytochemicals. Vero cells monolayer in 96-well plate was infected with RH strain of T. gondii and treated with concentrations of the EETC, Veratrine alkaloid, and clindamycin ranging from 1.56 to 200 µg/mL. MTT assay was conducted after 24 hours to evaluate the cytotoxicity and antiparasitic activities of the EETC. Four and 24 hours treatment models were adapted to assess the infection index and intracellular proliferation of T. RESULTS: The study revealed that the EETC had no cytotoxic effects on Vero cells with IC50 = 179 µg/mL, as compared to clindamycin (IC50 = 116.5 µg/mL) and Veratrine alkaloid (IC50 = 60.4 µg/mL). The EETC had good anti-toxoplasma activities with IC50 = 6.31 µg/mL in comparison with clindamycin (IC50 = 8.33 µg/mL) and Veratrine alkaloid (IC50 = 14.25 µg/mL). The EETC caused more than 70% and 80% reduction in infection index and intracellular proliferation in both treatment models, respectively. CONCLUSION: This in vitro study showed that the EETC contains promising phytochemicals effective against T. gondii and safe to the host cells.