Biocatalytic and Biological Activities of Cassia Occidentalis Mediated Silver Nanoparticles

Silver nanoparticles (AgNPs) were synthesized using the leaf extracts from Cassia occidentalis. The reactions were marked by a distinct colour change and the formation of AgNPs was monitored by measuring the UV-Vis spectra. The morphology and crystalline phase of the metal nanoparticles were determined using Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) with Energy Dispersive X-ray analysis (EDX) and Fourier-Transform-Infrared Spectroscopy (FTIR). The biocatalytic activity of the AgNPs against nitrifying compounds was determined using 4-nitrophenol and 4-nitroalanine. The biological screening involved analyzing the AgNPs effectiveness against gram-negative and gram-positive bacteria (Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus cereus and Enterococcus faecalis). The AgNPs displayed minimal inhibition at low concentrations ranging from 25 to 50 μg/mL. The toxicity of the AgNPs was tested using the brine shrimp assay and was observed as non-toxic with a nearly zero mortality rate. The AgNPs exhibited an antioxidant potential of 70.90 % inhibition at 1000 µg/mL. C. occidentalis is noted as a potential bioresource for synthesizing AgNPs with applications as antibacterial, antioxidant and biocatalytic agents. HIGHLIGHTS The addition of Ag nanoparticles to C. occidentalis plant extracts resulted in the formation of AgNPs-C. occidentalis complexes The AgNPs-C. occidentalis complexes were characterized using Transmission Electron Microscopy, Scanning Electron Microscopy with EDX, UV-Vis spectrophotometry and IR spectroscopy These AgNPs-C. occidentalis complexes were very stable and showed antibacterial activity against a range of tested gram-positive and gram-negative bacteria at low doses, thus proving to be good antimicrobial agents The AgNPs-C. occidentalis complexes showed higher scavenging activity than leaf extracts of C. occidentalis. The AgNPs have increased catalytic ability for the reduction of 4-nitrophenol and 4-nitroalanine after a 30 min time interval GRAPHICAL ABSTRACT

In-vitro Testing of Antioxidant, Anti-Parasite Activities, Cytotoxicity, and Chemical Evaluation of Abutilon Pannosum and Cassia Occidentalis Ethanolic Extracts

Aims: The aim of this study to detect Anti-giardia, antioxidant activities, cytotoxicity and evaluated the chemical constituent of ethanolic extracts of Abutilon pannosum and Cassia occidentalis. Study Design: Various standard methods were used to detect of bioactivity for ethanolic extracts of plants used in this study. Place and Duration of Study: This study was conducted in the laboratories of microbiology and parasitology and chemistry, the International University of Africa, Khartoum, Sudan, during May 2019. Methodology: The ethanolic extract of Abutilon pannosum and Cassia occidentalis was used as an anti-giardia and anti-oxidant in-vitro, and toxicity tests were performed using brine shrimp and MTT assay. Also, the compounds of the plants used were detected by the GCMS apparatus. Results: The ethanolic extracts of Abutilon pannosum showed high Anti-giardia activity (79%) in concentration (500 ppm) after 72 hours, whereas the activity of Cassia occidentalis extract showed (61%). The highest antioxidant activity of ethanolic extract of Cassia occidentalis was (68.7%), while it was weak in Abutilon pannosum ethanolic extract (45%) by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The results of cytotoxicity revealed that the ethanolic extracts are highly toxic to brine shrimp, but are not toxic to normal cell line (MTT). Chromatographic analysis using gas chromatography-mass spectroscopy (GCMS) showed good separation of compounds. GCMS detected 22 and14 important compounds in Abutilon pannosum and Cassia occidentalis extracts respectively. The common compound in both plant extracts is n-Hexadecanoic acid. This acid was reported as an antioxidant. Conclusion: This study revealed that the biological activities of Abutilon pannosum extracts showed high activities of Anti-giardia and antioxidants. Non-cytotoxic in the normal cell line was shown. Cassia occidentalis showed high activity of Anti-giardia and weak activity antioxidant.

GC-MS Analysis and Wound Healing Efficiency of Ethanolic Leaves Extract of Cassia occidentalis and Pithecellobium dulce in Wistar Albino Rats: Comparative Study

Aim: To identify and compare the bioactive compounds in the ethanolic leaves extracts of Cassia occidentalis and Pithecellobium dulce and to evaluate the wound healing efficiency in Wistar Albino rats. Study Design: The leaves ethanolic extracts was analysed by GC-MS and the extract was prepared in the form of a cream by ethanolic leaves extracts of C. occidentalis and P. dulce at 5% (w/v), 10% (w/v), and also in combination, a simple ointment base was developed with a composition of (1:1) Topical application of 5% (w/v) and 10% (w/v) was utilised in excision wound models. For excision wound models, the treatment duration was ten days. The day on which the wound was inflicted was designated as day '0'. Wound healing Activity: Excision wound Model: The animals were randomly separated into eight groups of six rats each: Group I: Control.; Group II: Standard group, treated with Framycetin sulfate cream (Soframycin, Aventis);. Group III: Treated with ethanolic extract of C. occidentalis (ELCO) (5% w/v); Group IV: Treated with ethanolic extract of C. occidentalis (ELCO) (10% w/v);Group V: Treated with ethanolic extract of P. dulce (ELPD)(5% w/v); Group VI: Treated with ethanolic extract of P. dulce (ELPD) (10 % w/v);Group VII: Treated with ethanolic extract of C. occidentalis and P. dulce (ELCO & ELPD 1:1) (5% w/v); Group VIII: Treated with ethanolic extract of C. occidentalis and P. dulce (ELCO & ELPD 1:1) (10 % w/v) till complete epithelization. Next dead space wound model and histology was studied. Place and Duration of Study: The GC-MS was carried out at Lab in Chennai. The extraction procedures were done at Department of Microbiology, Acharya Nagarjuna University, Guntur and treatment of wound healing activities were conducted at Ratnam Institute of Pharmacy in Nellore, Andhra Pradesh, India, and housed in the Department of Pharmacology between October to January 2016. Methodology: To study bioactive compounds, GC-MS was adopted, for wound healing activity: Excision wound Model, Dead space wound model and histology procedures was applied. Results: In the current study, ethanol leaves extract (EL) of Cassia occidentalis and Pithecellobium dulce were compared using GC-MS and their wound healing efficacy in wistar rats was examined. The GC-MS analysis of EL from both plants revealed 14-16 distinct bioactive phytochemical components with varying molecular weights and retention duration (RT). Excision and dead space wound models were utilised to assess the wound healing activities of EL extracts on rats. Wound concentration, full epithelialzation time, granulation, tissue weight, and hydroxyproline content were used to measure healing. In the excision wound model, the standard group (Framycetin sulphate cream) and group-VII (10% w/v; 1:1) combination EL treatment exhibited 98.5 ± 0.54 % and 98.4 ± 0.46 % wound healing activity, respectively. When compared to the control, the granulation tissue weight and hydroxyproline content in the dead space wound rose considerably. Histological examination revealed fewer inflammatory cells and more collagen, indicating a role in accelerating wound healing activity. Conclusion: The results of our investigation indicate unequivocally that ethanolic leaf extracts of these plant species are effective at encouraging wound healing. The 10% (ELCO+ELPD) tropical treatment drastically reduced the wound as compared to standard and also increased granulation and hydroxyproline content. However, it requires more clinical examination before being considered for wound therapy.

IMMUNOSTIMULATORY ACTIVITY OF AQUEOUS LEAVES EXTRACT OF CASSIA OCCIDENTALIS ON INNATE AND ADAPTIVE IMMUNE RESPONSE IN MICE

Different types of plants have been studied for their roles in immunostimulation and treatment of other diseases. The study aimed at evaluating the stimulatory activity of aqueous leaf extract of Cassia occidentalis on innate and adaptive immune responses in mice. Study mice were grouped into five and studied for 21 days; group I (control group) received normal saline. While the treatment groups: group II - V received aqueous leaf extract of C. occidentalis at different concentrations (250, 500, 1000, and 1500) mg/kg body weight (BW). The mice were sensitized and challenged with sheep red blood cells (SRBCs) on day 13th and 18th of the treatment, respectively. On the 19th, 20th, and 21st days, the animals’ delayed-type hypersensitivity (DTH) responses in each group were measured using a digital Vernier calliper, and humoral immune response was evaluated using the haemagglutination (HA) antibody technique. Blood samples were collected for Carbonic particle clearance index (K) and phagocytic index of Macrophage (α). Organ weight index (g/100g) was also recorded (α). The result showed a significant increase in the Carbonic particle clearance index (p=0.02) and phagocytic index of Macrophage (p<0.0001) in a dose-dependent manner. The study revealed a statistically significant difference in HA titres across the five groups, (p=0.02), also the DTH shows a statistically significant difference across the time (24 hours, 48 hours, 72 hours), Wilks’ Lambda = 0. 291, F = 15. 86, p = 0. 0001. The aqueous leaf extract of C. occidentalis has an immunostimulatory effect on innate and adaptive immunity.

Efficacy of Ricinus communis L., Cassia occidentalis L. and Bacillus thuringiensis against Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

Helicoverpaarmigera is one of the most destructive pests of field crops worldwide. The Study was designed to investigate through laboratory screening the insecticidal activity of Ricinus communis, Cassia occidentalis and Bacillus thuringiensis var. Kurstaki (Btk) and their combinations against 2nd larval instar of H. armigera. Five concentrations (4%, 6%, 8%, 10% and 12%) of each plant extract and (0.62, 1.25, 2.5, 5, and 10 mg/ml) of Bt were used in this experiment. Mortality (%) was recorded after 24,48,72 and 96 hrs post treatment. The results showed that R. communis, C. occidentalis and Btkhave insecticidal activity against 2nd instar larvae of H. armigera. The highest concentration (12%) caused 80% and 70% larval mortality after 96 hrs for R. communis and C. occidental respectively. The statistical analysis revealed that there is a significant differences between all treatments and control. Additionally, all combinations (plant/plant or plant/Bt) increased the mortality %. The results also clearly demonstrate that the R. communis are significantly more toxic than C. occidental where the LC50 values were 6.4 % for R. communis and 8.1% for C. occidental. On the other hand LC50 value for Bt was 0.41 mg/ml. The binary mixture of R. communis and C. occidental have a potentiation effect after 48 hrs. Regarding the mixture of Bt and plant extracts the results revealed that R. communis and Bt mixture induced a potentiation effect whereas C. occidental and Bt mixture induced a an additive effects.

Adsorption conceivable consequences of bio-sorbent got from cordia dichotoma (lasoda) and cassia occidentalis in the removal of methyl red dye

Sorption abilities of the sorbent material derived from Leaves and Barks of Lasoda (Cordia dichotoma) and Cassia Occidentalis were explored towards Methyl Red using synthetically prepared simulated wastewaters. Various factors such as initial dye concentration, contact time, adsorbent dosage, and the effect of temperature, which affects the adsorption, were evaluated. The equilibrium of adsorption was studied by Freundlich, Langmuir, Temkin, and Dubinin-Radushkevich isotherms. To identify the kinetics of the adsorption process, pseudo-first-order, pseudo-second-order, Weber and Morris intraparticle diffusion, Bangham’s pore diffusion, and Elovich equations were applied. The interference of fivefold excess of common anions and cations present in natural waters was studied. Cations like Ca2+, Mg2+, and Cu2+ have shown some interference but Fe2+ and Zn2+ have synergistically maintained the maximum extraction of the dye. The procedures developed have been successfully applied to some industrial effluent. The experimental data were suitable for the pseudo-First order kinetic model. The correlation coefficient (R2) and dimensionless separation factor (RL) values have confirmed that adsorption obeys Langmuir adsorption, indicating monolayer formation.

Comparative Efficacy of Antidermatophyte Creams Formulated from Ethanol Extracts of Cassia Occidentalis and Cassia Alata Leaves

Incidence of drug resistance by dermatophytes is a worldwide concern. Researchers search for natural and better drugs to arrest the ugly situation. This study aims to formulate antidermatophyte Creams with ethanol extracts of the leaves of cassia alata and cassia occidentalis against four selected dermatophytes namely Trichophyton mentagrophtes, Microsporum audounii, Epidermophyton floccosum and Malassezia furfur. Wistar rats were used in determining the efficacy of the formulated herbal creams by examining the biopsies of the skin of the infected rats for presence of fungal hyphae, tissue destruction, Inflammation of the skin, absence of sebaceous gland and also absence of hair follicles. Dekeratinization of the skin layer was also measured. From the result obtained, it was observed that the efficacy of the creams is concentration dependent. Cassia alata cream formulation was particularly effective against three of the dermatophytes namelyTrichophyton mentagrophtes (30.71± 0.51 µm) Microsporum audounii (36.17± 0.59 µm), and Epidermophyton floccosum (37.49± 0.18 µm) while Cassia occidentalis cream was effective against Malassezia furfur (33.22± 0.16 µm). The results showed that the plants ethanol extracts can be utilized in the management of dermatophytosis when formulated as a cream for topical use.