Development of an allele-specific PCR (AS-PCR) method for identifying high-methyl eugenol-containing purple Tulsi (Ocimum tenuiflorum L.) in market samples.

BACKGROUND: Ocimum tenuiflorum L. is a highly traded medicinal with several therapeutic values. Green Tulsi and purple Tulsi are two subtypes in O. tenuiflorum and both have the same medicinal properties. Recent reports have revealed that purple Tulsi contains higher quantities of methyl eugenol (ME), which is moderately toxic and potentially carcinogenic. Therefore, we developed an allele-specific PCR (AS-PCR) method to distinguish the green and purple Tulsi. METHODS AND RESULT: Using the green Tulsi as a reference, 12 single nucleotide polymorphisms (SNPs) and 10 insertions/deletions (InDels) were identified in the chloroplast genome of the purple Tulsi. The C > T SNP at the 1,26,029 position in the ycf1 gene was selected for the development of the AS-PCR method. The primers were designed to amplify 521 bp and 291 bp fragments specific to green and purple Tulsi, respectively. This AS-PCR method was validated in 10 accessions from each subtype and subsequently verified using Sanger sequencing. Subsequently, 30 Tulsi powder samples collected from the market were subjected to molecular identification by AS-PCR. The results showed that 80% of the samples were purple Tulsi, and only 3.5% were green Tulsi. About 10% of the samples were a mixture of both green and purple Tulsi. Two samples (6.5%) did not contain O. tenuiflorum and were identified as O. gratissimum. CONCLUSION: The market samples of Tulsi were predominantly derived from purple Tulsi. The AS-PCR method will be helpful for quality control and market surveillance of Tulsi herbal powders.

Chemical composition and mosquitocidal properties of essential oil from Indian indigenous plants Ocimum tenuiflorum L. and Ocimum americanum L. against three vector mosquitoes.

Mosquitoes stand out as the most perilous and impactful vectors on a global scale, transmitting a multitude of infectious diseases to both humans and other animals. The primary objective of the current research was to assess the effectiveness of EOs from Ocimum tenuiflorum L. and Ocimum americanum L. in controlling Anopheles stephensi Liston. Culex quinquefasciatus Say and Aedes aegypti L. mosquitoes. The larvae, pupae and eggs of the mosquitoes were exposed to four different concentrations (6.25-50 ppm). The tested EOs resulted in >99-100 % mortality at 120 h for the eggs of all examined mosquito species. It also showed robust larvicidal and pupicidal activity with LC50 and LC90 values of 17-39, 23-60 ppm and 46-220, and 73-412 ppm against Aedes, Culex and Anopheles mosquito species, respectively, at 24 h of treatment. The Suitability Index or Predator Safety Factor demonstrated that the EOs extracted from O. tenuiflorum L. and O. americanum L. did not cause harm to P. reticulata, D. indicus (water bug), G. affinis and nymph (dragonfly). GC-MS analysis identified the major probable constituents of the oil, including Phenol, 2-Methoxy-4-(1-Propenyl)- (28.29 %); 1-Methyl-3-(1'-Methylcyclopropyl) Cyclopentene (46.46 %); (E,E,E)-3,7,11,15-Tetramethylhexadeca-1,3,6,10,14-Pentaene (18.91 %) and 1,3-Isobenzofurandione, 3a,4,7,7a-Tetrahydro-4,7-Dimethyl (33.02 %). These constituents may play a significant role in the mosquitocidal activity of the oil. The same results were identified in the formulation prepared from the EOs. This marks the first report confirming the successful utilization of EOs derived from O. tenuiflorum L. and O. americanum L. in mosquito population control initiatives.

Mitigation of arsenic poisoning induced oxidative stress and genotoxicity by Ocimum gratissimum L.

Arsenic toxicity is a major problem across the world due to geogenic activity and has been supposed to generate free radicals and genotoxicity among the arsenic-poisoned population. There is a need to find suitable free radical quenching compounds for the arsenic-induced free radical-affected population. In the present study, Na3AsO3- induced oxidative stress and genotoxicity were evaluated in Oryctolagus cuniculus L, and quenching competency of Ocimum species was examined by applying enzymatic and non-enzymatic in vitro tests, comet assay, and Random Amplified Polymorphic Deoxyribonucleic acid - Polymerase Chain Reaction (RAPD-PCR) methods. In the present study, oxidative damage due to Na3AsO3 intoxication in O. cuniculus L has been confirmed followed by substantive genotoxicity, and in a further study, it has also been reported that the extract of O. gratissimum L lowers the oxidative stress in experimental animals confirmed by a decrease in Malondialdehyde (MDA) 4.78 ± 0.05 (nmol/mg protein), and an increase in Glutathione (GSH) 2.87 ± 0.50 (µmoles/mg proteins), Superoxide Dismutase (SOD) 1.78 ± 0.03(Units/mg protein), Catalase (CAT) 2.72 ± 0.02 (µmoles of H2O2 consumed/min/mg proteins) and Glutathione peroxidase (GPX) 7.43 ± 0.01 (µg of glutathione utilized/min/mg protein). A positive impact of extract of O. gratissimum L on protection of genotoxicity has been also confirmed by Random Amplified Polymorphic DNA (RAPD) based reduction in polymorphic bands of Deoxyribonucleic acid (DNA) from 6.5 to 3.16 and comet assay-based increase in head DNA % (87.86 ± 1.58), tail moment (1.07 ± 0.27) and decrease in tail DNA % (12.13 ± 1.58) & tail length (8.2 ± 1.46) at 5% P in lymphocytes. A significant level reduction in free radicals and reduction in DNA polymorphism has proved the competency of test material for the development of suitable antidotes against arsenicosis.

Insecticidal property of Ocimum essential oil embedded polylactic acid packaging films for control of Sitophilus oryzae and Callosobruchus chinensis.

Naturally derived essential oils (EOs) from Ocimum basilicum (OB) and Ocimum gratissimum (OG) were incorporated in polylactic acid (PLA) matrix with concentration of 2.5 % and 5 % respectively by solvent casting method, with an aim to develop insecticidal polymer pouches (insect proof polymer pouches). The major components following GC-MS analysis revealed the presence of EOs as estragole (69.998 %) in OB and thymol (29.843 %) in OG. The developed films with EOs showed remarkable insecticidal activity with 100 % and 80 % mortality rate for OB and OG respectively at 2.5 % concentration against rice weevil (Sitophilus oryzae). While, both OB and OG EOs observed 100 % mortality against pulse beetle (Callosobruchus chinensis) after 96 h exposure in presence of grain conditions and the insecticidal efficacy was equivalent to the commercial hermetic bags (CSB). The physical property of film showed increase in the film thickness in EO-PLA films whereas the mechanical properties of EO-PLA films matrix have increased the polymer flexibility with decreasing tensile strength (TS) and Youngs modulus (EM) as TS OB 5 %, OG 5 % - 15.64 MPa, 17.66 MPa respectively. While, water vapour transmission rate (WVTR) was reduced to 0.015 g/m2.hr, optical characteristics showed slight significant change in colour and FTIR spectra exhibited a change in -OH stretching compared to blank PLA matrix. The surface morphology of the blank and EO embedded PLA films visualized by SEM depicted a complete homogenization of essential oils in the PLA matrix.

Differential impacts of interactions between Serendipita indica, Chlorella vulgaris, Ulva lactuca and Padina pavonica on Basil (Ocimum basilicumL.).

Plant biostimulants (PBs) are used globally to increase crop yield and productivity. PBs such as (Serendipita indica) or algal extracts stimulate and accelerate plant physiological processes. The physiological, ecological, and biochemical effects of (Serendipita indica) or algal extracts individually and in combination on basil plant (Ocimum basilicum L.) were investigated. Macroalgae samples were collected from Abu Qir, Alexandria, Egypt. The growth parameters, chlorophyll index, and biochemical composition of basil were analyzed at 90th day. The (Chlorella vulgaris) + (Serendipita indica) (MI + F) treatment increased chlorophyll index by 61.7% (SPAD) compared to control. (Chlorella vulgaris) had the highest growth hormones, including GA3 at 158.2 ppb, GA4 at 149.1 ppb, GA7 at 142.6 ppb, IAA at 136.6 ppb, and TC at 130.9 ppb, while (Ulva lactuca) had the lowest. The MI + F treatment yielded the highest essential oil and antioxidant values. Treatment with (Chlorella vulgaris) increased S. indica colonization by 66%. In contrast, Ulva lactuca and (Padina Pavonica) inhibited S. indica colonization by 80% and 40%, respectively. (Ulva lactuca) and (Padina Pavonica) inhibited S. indica colonization by 80% and 40%, respectively. Combined treatments had a greater influence on basil performance than the individual treatments. The evidence of synergistic/additive benefits to plants performance due to the interactive effects of (Chlorella vulgaris) and (Serendipita indica) had been studied. Complementary modes of action between (Chlorella vulgaris) and (Serendipita indica), through their components newly emerging properties on basil, may explain observed synergistic effects. This study explores the potential of microbial-algal interactions, particularly (Chlorella vulgaris) and (Serendipita indica), as innovative plant biostimulants. These interactions demonstrate positive effects on basil growth, offering promise for more effective microbial-based formulations to enhance crop productivity and sustainability in agriculture. These novelties will help create a second generation of PBs with integrated and complementary actions.

Low-temperature perception and modulations in Ocimum basilicum commercial cultivar CIM-Shishir: Biosynthetic potential with insight towards climate-smart resilience.

The newly released interspecific hybrid variety CIM-Shishir, resulting from a cross between Ocimum basilicum and Ocimum kilimandscharicum claims to be a multicut, lodging resistant, cold tolerant, high essential oil yielding with linalool rich variety. It has a purple-green stem and has a unique feature and advantage of better survival in the winter season than other O. basilicum varieties, illustrating its physiological mechanisms for cold tolerance. In this study, we subjected both the CIM-Shishir variety and a control plant to cold stress to investigate the impact of low temperatures on various physiological, trichome developments, secondary metabolite constitution aspects related to essential oil production, and gene expression. The analysis revealed a significantly higher density and altered morphology of trichomes on the leaf surface of the variety subjected to low temperatures, indicating its adaptation to cold conditions. Furthermore, when comparing the treated plants under low-temperature stress, it was observed that the relative electrolyte leakage and Malondialdehyde (MDA) contents substantially increased in the control in contrast to the CIM-Shishir variety. This finding suggests that CIM-Shishir exhibits superior cold tolerance. Additionally, an increase in proline content was noted in the variety exposed to low temperatures compared to the control. Moreover, the chlorophyll and anthocyanin content gradually increased with prolonged exposure to low-temperature stress in the newly developed variety, indicating its ability to maintain photosynthetic capacity and adapt to cold conditions. The activities of superoxide dismutase (SOD) also increased under low-temperature conditions in the CIM-Shishir variety, further highlighting its cold tolerance behaviour. In our research, we investigated the comprehensive molecular mechanisms of cold response in Ocimum. We analyzed the expression of key genes associated with cold tolerance in two plant groups: the newly developed hybrid variety known as CIM-Shishir Ocimum, which exhibits cold tolerance, and the control plants susceptible to cold climates that include WRKY53, ICE1, HOS1, COR47, LOS15, DREB5, CBF4, LTI6, KIN, and ERD2. These genes exhibited significantly higher expression levels in the CIM-Shishir variety compared to the control, shedding light on the genetic basis of its cold tolerance. The need for climate-smart, resilient high-yielding genotype is of high importance due to varied climatic conditions as this will hit the yield drastically and further to the economic sectors including farmers and many industries that are dependent on the bioactive constituents of Ocimum.

Voltammetric techniques for low-cost on-site routine analysis of thymol in the medicinal plant Ocimum gratissimum.

The composition of essential oils varies according to culture conditions and climate, which induces a need for simple and inexpensive characterization methods close to the place of extraction. This appears particularly important for developing countries. Herein, we develop an analytical strategy to determine the thymol content in Ocimum Gratissimum, a medicinal plant from Benin. The protocol is based on electrochemical techniques (cyclic and square wave voltammetry) implemented with a low cost potentiostat. Thymol is a phenol derivative and was directly oxidized at the electrode surface. We had to resort to submillimolar concentrations (25-300 µM) in order to minimize production of phenol oligomers that passivate the electrode. We worked first on two essential oils and realized that in one of them the thymol concentration was below our detection method. These results were confirmed by gas chromatography - mass spectrometry. Furthermore, we optimized the detection protocol to analyze an infusion made directly from the leaves of the plant. Finally, we studied whether the cost of the electrochemical cell may also be minimized by using pencil lead as working and counter electrodes.

Isolation and Identification of Endophytic Bacterial Isolates from the Leaves, Roots, and Stems Parts of Artemisia annua, Moringa oleifera, and Ocimum lamiifolium Plants.

Medicinal plants are known to harbor diverse species of endophytic bacteria which are known for secretion of beneficial secondary metabolites, like enzymes and antimicrobial compounds. The present study aimed to isolate, characterize, and identify the endophytic bacteria isolates from Artemisia annua, Moringa oleifera, and Ocimum lamiifolium plants. Certain endophytic bacterial isolates were screened. Phosphate and Zinc solubilization were performed for newly obtained isolates. The 16S rRNA gene sequencing was performed for RPAAI-8 isolate. Data were analyzed. Our study showed that endophytic bacterial isolates were recognized to be Bacillus cereus, B. subtilis, Citrobacter freundii, Enterobacter asburiae, E. cloacae, E. kobei, E. ludwigii, Enterococcus faecium, and Pseudomonas monteilli. From among these differentiated endophytic bacterial isolates, Enterobacter species are the most frequently obtained isolates. These bacterial isolates were shown 99.77% sequence similarity to Enterobacter ludwigii EN-119T (JTLO01000001) using 16S rRNA gene sequencing. This isolate was designated as Enterobacter sp. RPAAI-8. This isolate was able to employ selected cheap and cost-effective agro wastes as a carbon source. This cheap agro waste utilization by these Enterobacter species could be the first report. In conclusion, the present isolates are found to be employed for plant growth promotion and solubilizing insoluble phosphate and zinc. Before this time, most of the recent isolates were not identified from these medicinal plants. The ethyl acetate extract of the isolates also showed inhibitory activity against selected test pathogens.

Chemical compositions, antioxidant, antimicrobial, and mosquito larvicidal activity of Ocimum americanum L. and Ocimum basilicum L. leaf essential oils.

BACKGROUND: Ocimum americanum L. (O. americanum) and Ocimum basilicum L. (O. basilicum) are highly valued aromatic medicinal plants. Their leaves are widely used as spices in traditional cuisine. Their essential oils (EOs) are extensively used in food, cosmetic, and pharmaceutical industries. This study aimed to investigate the main chemical profiles of O. americanum and O. basilicum leaf EOs and assess their effects on antibacterial, antioxidant, and larvicidal properties. METHODS: EOs were extracted from the leaves of O. basilicum and O. americanum using steam distillation in a Clevenger-type apparatus. The chemical constituents of the EOs were analyzed using gas chromatography-mass spectrometry. 2,2-Diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), and metal-chelating techniques were used to assess the free-radical scavenging capability of the oils. The extracted oils were also tested for their antibacterial activities via a disk-diffusion test and the broth microdilution method. Furthermore, the mosquito larvicidal (Aedes aegypti) activity was tested using standard protocols. RESULTS: Camphor (33.869%), limonene (7.215%), longifolene (6.727%), caryophyllene (5.500%), and isoledene (5.472%) were the major compounds in O. americanum leaf EO. The EO yield was 0.4%, and citral (19.557%), estragole (18.582%) camphor (9.224%) and caryophyllene (3.009%) were the major compounds found among the 37 chemical constituents identified in O. basilicum oil. O. basilicum exhibited a more potent antioxidant activity in DPPH, FRAP, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid tests than O. americanum. The zones of inhibition and minimum inhibitory concentration of the oils in the microdilution and disk diffusion methods were 8.00 ± 0.19 mm to 26.43 ± 2.19 mm and 3.12-100 µg/mL, respectively. At 400 ppm, O. basilicum and O. americanum EOs demonstrated larvicidal activity, with mortality ratios of 73.60% ± 0.89% and 78.00% ± 1.00%, respectively. Furthermore, after 30 min of exposure to O. americanum and O. basilicum EOs, the larval death rates were 73.60% ± 0.89% and 78.00% ± 1.00%, respectively. CONCLUSIONS: The findings revealed that the EOs extracted from the leaves of O. basilicum and O. americanum exhibited reasonable antioxidant, antibacterial, and mosquito larvicidal potentials, and can be used as alternative medicine for the treatment of human health and larvicidal mosquito control.

Development of gelatin/agarose active coatings functionalized with Ocimum gratissimum L. essential oil for enhancing storability of 'Booth 7' avocado.

Novel active coating from gelatin/agarose (GA) functionalized with Ocimum gratissimum L. essential oil (OGO) had been developed as a medium to evaluate their properties before being applied for avocado preservation. The resultant coating films showed enhanced mechanical, water-barrier, bactericidal, antioxidant, and UV-shielding properties by adding OGO. The best tensile strength (2.91 MPa) and flexibility (45.82 %) was found in the GA film containing 5 % (w/w) of OGO (GA-OGO-5). Furthermore, this coating formulation presented moderate antibacterial activities against Listeria, Pseudomonas, Salmonella, and Escherichia. The GA-OGO-5 coating film also divulged the highest hydrophobicity and adequate antioxidant function (30.75 µg/mL) and thus, was chosen to coat on 'Booth 7' avocados by dipping method. The GA-OGO-5 coating layers were to be efficient to decline the respiration rate of avocado during 6-day storage at 25 °C and 64 %RH. Peel color, weight loss (5.22 %), total soluble solids (8.14 %), and solution pH (6.79) at the end of storage also indicated that the GA-OGO-5 coating presented the best effectiveness for enhancing the storability of avocado as compared to uncoated and GA-treated fruit. Therefore, the GA-OGO coating has been considered as an alternative post-harvest technique to enhance the avocado storability and could be further commercialized for industry application.

A comparison of high- and low-resolution gas chromatography-mass spectrometry for herbal product classification: A case study with Ocimum essential oils.

INTRODUCTION: Selection of marker compounds for targeted chemical analysis is complicated when considering varying instrumentation and closely related plant species. High-resolution gas chromatography-mass spectrometry (GC-MS), via orbitrap detection, has yet to be evaluated for improved marker compound selection. OBJECTIVE: This study directly compares high- and low-resolution GC-MS for botanical maker compound selection using Ocimum tenuiflorum L. (OT) and Ocimum gratissimum L. (OG) for botanical ingredient authentication. METHODS: The essential oils of OT and OG were collected via hydrodistillation before untargeted chemical analysis with gas chromatography coupled to single-quadrupole (GC-SQ) and orbitrap (GC-Orbitrap) detectors. The Global Natural Products Social Molecular Networking (GNPS) software was used for compound annotation, and a manual search was used to find the 41 most common Ocimum essential oil metabolites. RESULTS: The GC-Orbitrap resulted in 1.7-fold more metabolite detection and increased dynamic range compared to the GC-SQ. Spectral matching and manual searching were improved with GC-Orbitrap data. Each instrument had differing known compound concentrations; however, there was an overlap of six compounds with higher abundance in OG than OT and three compounds with a higher abundance in OT than OG, suggesting consistent detection of the most variable compounds. An unsupervised principal component analysis (PCA) could not discern the two species with either dataset. CONCLUSION: GC-Orbitrap instrumentation improves compound detection, dynamic range, and feature annotation in essential oil analysis. However, considering both high- and low-resolution data may improve reliable marker compound selection, as GC-Orbitrap analysis alone did not improve unsupervised separation of two Ocimum species compared to GC-SQ data.

Antimicrobial Effect of Ocimum gratissimum L. Essential Oil on Shewanella putrefaciens: Insights Based on the Cell Membrane and External Structure.

The main objective of this study was to assess the in vitro antibacterial effectiveness of Ocimum gratissimum L. essential oil (OGEO) against Shewanella putrefaciens. The minimum inhibitory concentration and minimum bactericidal concentration of OGEO acting on S. putrefaciens were both 0.1% and OGEO could inhibit the growth of S. putrefaciens in a dose-dependent manner. The restraint of the biofilm growth of S. putrefaciens was found in the crystal violet attachment assay and confocal laser scanning microscopy. The disruption of cell membranes and exudation of contents in S. putrefaciens with OGEO treatment were observed by scanning electron microscopy, hemolysis and ATPase activity. The results demonstrated that OGEO had a positive inhibitory effect on the growth of S. putrefaciens, which primarily developed its antibacterial function against S. putrefaciens by disrupting the formation of biofilms and cell membranes. This study could provide a new method of inhibiting the spoilage of food in which the dominant spoilage bacteria are S. putrefaciens.

Gelling and rheological properties of a polysaccharide extracted from Ocimum album L. seed.

In this study, a new polysaccharide was isolated from Ocimum album L. seed (OA), and its physicochemical and rheological properties were investigated. Ocimum album polysaccharide (OAP) was an acidic heteropolysaccharide with a molecular weight of 1935 kDa, and it was composed of five types of sugars: mannose (32.95 %), glucose (27.57 %), galactose (19.29 %), rhamnose, (15.96 %) and galacturonic acid (4.23 %). According to the results obtained from Huggins and Kraemer equations, the intrinsic viscosity was 6.9 dL/g in distilled water. The OAP solutions at a concentration between 0.1 and 1.5 %, showed shear-thinning behavior, and the Herschel-Bulkley and Cross models exhibited a high ability to describe the flow behavior of OAP solutions. The apparent viscosity of 1 % OAP solution was decreased in the presence of different concentrations of NaCl (0.1, 0.3, and 0.5 M), at different pHs (3-11), and in temperatures between 5 and 100 °C. Also, the pseudoplastic behavior was observed in all samples. In OAP solutions (0.1-1.5 %), the up and down curves in the shear stress-shear rate diagram did not coincide, which indicated time-dependent (thixotropic) behavior. Although, the thixotropic properties of 1 % OAP solution were weakened with adding NaCl (0.1-0.5 M) and at different pH (3-11). The results obtained from the dynamic oscillatory test showed that the OAP solutions at concentrations higher than 0.1 % had a gel-like behavior, and the viscoelastic moduli (G' and G″) were weakened in the presence of salt and with a change in pH. Also, in the temperature sweep test, the 1 % solution showed the behavior of thermally irreversible gels.

In Vitro and Molecular Docking Evaluation of Larvicidal Effects of Essential Oils of Five Aromatic Plants on the Fall Armyworm Spodoptera frugiperda JE. Smith (Lepidoptera: Noctuidae) from Ivory Coast.

Faced with the serious consequences resulting from the abusive and repeated use of synthetic chemicals, today rethinking crop protection is more than necessary. It is in this context that the essential oils of the Lamiaceae Ocimum gratissimum and Ocimum canum, the Poaceae Cymbopogon citratus and nardus and a Rutaceae Citrus sp. of known chemical compositions were experimented. The evaluation of the larvicidal potential of the essential oils was done by the method of topical application of the test solutions, on the L1-L2 stage larvae from the first generation of S. frugiperda obtained after rearing in an air-conditioned room. Lethal concentrations (LC10 , LC50 and LC90 ) were determined after 48 h. After assessing the larvicidal potential of essential oils, molecular docking was carried out to study protein-ligand interactions and their propensity to bind to insect enzyme sites (AChE). The essential oil of O. gratissimum was the most effective with the lowest lethal concentrations (LC10 =0.91 %, LC50 =1.91 % and LC90 =3.92 %). The least toxic oil to larvae was Citrus sp. (LC10 =5.44 %, LC50 =20.50 % and LC90 =77.41 %). Molecular docking revealed that p-cymene and thymol from O. gratissimum essential oil are structurally similar and bind to the AChE active site via predominantly hydrophobic interactions and a H-bond with Tyr374 in the case of thymol. The essential oil of O. gratissimum constitutes a potential candidate for the development of biological insecticides for the fight against insect pests and for the protection of the environment.

Chemical and Olfactory Analysis of the Volatile Fraction of Ocimum gratissimum Concrete from Madagascar.

The chemical composition of the volatile fraction of Ocimum gratissimum concrete (romba) from Madagascar has been determined for the first time by GC/MS and GC-FID. A methyl cinnamate chemotype has been determined for this material, along with a set of compounds typical in essential oils and extracts from plants of the Ocimum genus. Variability was mostly observed on terpenes and terpenoids components. GC-O-MS was also used for a sensory evaluation of this material performed by a master perfumer. The chemical composition of this O. gratissimum extract was then compared with literature data to assess subtle differences between chemotypes of the same species and other species of the same genus within natural variability. A mapping illustrates the occurrence of the cinnamate chemotype in Eastern Africa, India and now Madagascar, while other origins generally present eugenol, thymol, camphor, or linalool chemotypes.

Formulation and Evaluation of Quality Parameters of Effervescent Granules from the Potent Antioxidant between Two Variants of the Adaptogenic Herb Ocimum tenuiflorum L.

Ocimum tenuiflorum L. is found throughout semitropical and tropical parts of Southeast Asia. In Nepal, O. tenuiflorum L. is popular with two variants: Krishna Tulsi consisting of purple-colored leaves and Sri Tulsi consisting of green-colored leaves. O. tenuiflorum L. is considered the queen of herbs and is a traditionally and clinically proven medicinal herb for its application and efficacy. However, no commercial pharmaceutical preparations of O. tenuiflorum L. are available using effervescent vehicles. Therefore, the present study aimed to compare the antioxidant activity of leaves from the two varieties of O. tenuiflorum L. and formulate and evaluate the quality parameters of effervescent granules of the potent extract. The antioxidant activity of O. tenuiflorum L. ethanolic extracts was evaluated by DPPH radical scavenging assay at 1, 10, and 100 µg/mL concentrations, where ascorbic acid was used as the positive control. The antioxidant activity of purple-leafed O. tenuiflorum L. was found to be higher than that of green-leafed O. tenuiflorum L. Therefore, effervescent granules of the ethanolic extract of purple-leafed O. tenuiflorum L. were formulated using the pharmaceutical excipients tartaric acid, citric acid, and sodium bicarbonate and the quality parameters of the granules were evaluated. The formulated granules met the quality parameters assessed from the angle of repose, bulk density, tapped density, Carr's Index, Hausner's ratio, effervescent cessation time, and stability studies. Thus, the formulated effervescent granules of O. tenuiflorum L. can be used for therapeutic purposes or as a functional food.

Characteristic features and comparative analysis of essential oil composition of selected genus of Ocimum sanctum L. through GC-MS.

BACKGROUND & OBJECTIVES: The main aim of this research is to provide literature on the Ocimum plant, and to know the significance of the Ocimum species carried out by pharmacognostic study and experimental design for GC-MS. Ocimum genus are very important for their therapeutic potential among the most important aromatic herbs. METHODS: Extreme attention has been put on literature reports in which the utilization of tulsi and their pharmacognostic study has been done by performing morphological and microscopic leaf experimental design and by using essential oil through the GC-MS instrumentation method. RESULTS: The utilization of these characteristics would be important for the drug discovery scientist to develop a specific formulation of the crude drug, which will be a magical therapeutic agent in the future, with many advantages. GC-MS chromatogram of Ocimum sanctum, Ocimum canum, and Ocimum gratissimum oil showed major peaks and has been identified after comparison of the mass spectra with the NIST library, indicating the presence of three phytocomponents. From the results, the GC-MS study suggested that anethole which is well reported antimicrobial compound is more in O. canum (2.66%) in comparison to O. sanctum (1,28%) but absent in O. gratissimum. The results indicated that the antimicrobial activity is more in O. canum due to the presence ofa high amount of anethole in comparison to O. gratissimum and O. sanctum. INTERPRETATION & CONCLUSION: The result revealed that O. canum has a microscopic character that can be identified by the characteristic GC MS analysis of extracts to distinguish between different species of the ocimum plant.

Evaluation of the mosquito larvicidal potential and comparative assessment of the juice of Lantana camara Linn and Ocimum gratissimum Linn.

In the present study, the larvicidal efficacy of the juices of the weeds Lantana camara Linn (L. camara) and Ocimum gratissimum Linn (O. gratissimum) was evaluated against the larvae of the malaria vectors Aedes aegypti, Anopheles subpictus and Culex quinquefasciatus. The freshly prepared juices of leaves were prepared by grinding them and diluting them at concentrations of 25, 50, 75, and 100 ppm. Twenty larvae of each species were introduced in different sterile Petri dishes in aqueous media under a controlled environment for the assessment of biological activity. The larvicidal activity of both juices was evaluated at 6, 12 and 24 h post-exposure time points by observing the movement of each larva. The obtained data were subjected to probit analysis to determine the lethal concentrations that kill 50% and 90% (LC50 and LC90) of the treated larvae. The results revealed a noticeable larvicidal activity following 24 h of exposure. The juice of L. camara leaves exhibited an LC50 range of 47.47-52.06 ppm and an LC90 range of 104.33-106.70 ppm. Moreover, for the juice of O. gratissimum leaves, the LC50 range was 42.94-44.91 ppm and the LC90 range was 105.11-108.66 ppm. Taken together, the results indicate that the juices of L. camara and O. gratissimum leaves may be useful as effective, economical and eco-friendly larvicidal agents. However, additional studies are needed to explore the bioactive components of the weeds that exhibit larvicidal activity along with their mode of action.

Sustainable fabrication of hybrid silver-copper nanocomposites (Ag-CuO NCs) using Ocimum americanum L. as an effective regime against antibacterial, anticancer, photocatalytic dye degradation and microalgae toxicity.

In this study, a sustainable fabrication of hybrid silver-copper oxide nanocomposites (Ag-CuO NCs) was accomplished utilizing Ocimum americanum L. by one pot green chemistry method. The multifarious biological and environmental applications of the green fabricated Ag-CuO NCs were evaluated through their antibacterial, anticancer, dye degradation, and microalgae growth inhibition activities. The morphological features of the surface functionalized hybrid Ag-CuO NCs were confirmed by FE-SEM and HR-TEM techniques. The surface plasmon resonance λmax peak appeared at 441.56 nm. The average hydrodynamic size distribution of synthesized nanocomposite was 69.80 nm. Zeta potential analysis of Ag-CuO NCs confirmed its remarkable stability at -21.5 mV. XRD and XPS techniques validated the crystalline structure and electron binding affinity of NCs, respectively. The Ag-CuO NCs demonstrated excellent inhibitory activity against Vibrio cholerae (19.93 ± 0.29 mm) at 100 µg/mL. Anticancer efficacy of Ag-CuO NCs was investigated against the A549 lung cancer cell line, and Ag-CuO NCs exhibited outstanding antiproliferative activity with a low IC50 of 2.8 ± 0.05 µg/mL. Furthermore, staining and comet assays substantiated that the Ag-CuO NCs hindered the progression of the A549 cells and induced apoptosis as a result of cell cycle arrest at the G0/G1 phase. Concerning the environmental applications, the Ag-CuO NCs displayed efficient photocatalytic activity against eosin yellow degradation up to 80.94% under sunlight irradiation. Microalgae can be used as an early bio-indicator/prediction of environmental contaminants and toxic substances. The treatment of the Ag-CuO NCs on the growth of marine microalgae Tetraselmis suecica demonstrated the dose and time-dependent growth reduction and variations in the chlorophyll content. Therefore, the efficient multifunctional properties of hybrid Ag-CuO NCs could be exploited as a regime against infective diseases and cancer. Further, the findings of our investigation witness the remarkable scope and potency of Ag-CuO NCs for environmental applications.

Sub-chronic toxicity of the aqueous leaf extract of Ocimum lamiifolium Hochst. ex Benth on biochemical parameters and histopathology of liver and kidney in rats: in vivo and in- silico toxicity studies.

BACKGROUND: The aerial part of Ocimum lamiifolium is commonly used in Ethiopian traditional medicine. Although this plant is mostly used in traditional medicine, its safety profile has not been documented yet. The aim of this study was to assess the sub-chronic toxicity of O. lamiifolium aqueous extract in rats and to determine the toxicity profile of GC-MS identified bioactive compounds obtained from essential oil of O. lamiifolium using in silico toxicity methods. METHODS: Eighty rats (40 male and 40 female) were randomly assigned to four groups of ten rats per sex/group. For 90 days, Groups I-III received 200, 400, and 600 mg/kg bw of aqueous extract of O. lamiifolium, respectively. Distilled water was given to Group IV (control). Clinical observations, food intake, and rat weight were all recorded during the experiment. In addition, several biochemical parameters, organ weight, and histology of the liver and kidney were all evaluated. For the in-silico toxicity study, GC-MS identified bioactive compounds in O. lamiifolium essential oil were obtained from published articles. The compounds two-dimensional structures were constructed using Chemdraw. The two-dimensional structures were converted into a simplified molecular input line entry system (SMILES) using the Swiss ADMET web tool. Furthermore, the toxicity parameters were predicted using the ProTox II server. RESULTS: The administration of an aqueous extract of O. lamiifolium leaves significantly (p < 0.05) reduced the test animals' food intake and body weight gain. In the high dose (600 mg/kg bw) treated group, the serum alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels were significantly increased (p < 0.05). In female rats given 600 mg/kg bw of O. lamiifolium, the levels of serum urea were also increased. In addition, rats given 600 mg/kg bw had significantly lower blood glucose levels than the control group (p < 0.05). Doses up to 400 mg/kg bw didn't bring a significant change to the histology of the liver. However, in the high dose (600 mg/kg bw) treated group, some female rats' livers showed mild sinusoidal and central vein dilatation, as well as parenchymal necrosis. our findings showed that all compounds derived from the essential oil of O. lamiifolium showed no mutagenicity or cytotoxicity. However, 30% of the compounds tested were hepatotoxic, 20% carcinogenic, and 20% immunotoxin. CONCLUSION: Our findings showed that oral administration of O. lamiifoliums aqueous extract up to a dose of 400 mg/kg bw is not toxic. However, high-dose (600 mg/kg bw) significantly affected the food consumption and weight gain of the experimental rats and the serum concentration of some liver and kidney enzymes were also significantly increased. Additionally, a considerable proportion of the tested compounds were predicted to be hepatotoxic, carcinogenic and immunotoxin. Furthermore, before employing O. lamiifolium preparations as drugs, a chronic toxicity research on the essential oil as well as its components that exhibited toxicity in the in-silico toxicity study is needed. Finally, use high doses of O. lamiifolium leaves with caution.