In vitro callus induction and antioxidant activity of Rauwolfia vomitoria Afzel. (Apocynaceae)

Background: Rauwolfia vomitoria Afzel. (Apocynaceae) is a medicinal plant valued for its antipsychotic effect and used in various herbal preparations.Objectives: This study was designed to develop a protocol for callus initiation in Rauvolfia vomitoria and to investigate the antioxidant activity of the wild plant and the leaf-derived callus of the plant.Materials and methods: Callus initiation of the leaf explants was achieved on  Murashige and Skoog’s medium fortified with α-naphthaleneacetic acid (NAA), 2, 4-dichlorophenoxyacetic acid (2,4- D) and 6-benzylamino purine (BAP). Methanol extracts of leaf and root of the wild plant and callus were analyzed for their  antioxidant activity using 2, 2, diphenyl-1-picryl hydrazyl (DPPH) for the radical scavenging activity (RSA) and Folin Ciocalteau spectrophotometric method for the total phenolic content (TPC).Results: Of the various plant growth hormones employed in this study for callus establishment, 1.0 mg/L NAA + 4.0 mg/L BAP induced maximum callus formation of 83.0%, while 1.0 mg/L 2,4-D gave 20.0% formation. Leaf explants placed in the dark phase produced whitish, friable calli. The highest antioxidant activity was obtained from the root extract of the wild plant with IC50 values of 3.56±1.67 and 429.72±19.83 μgGAE/g for RSA and TPC, respectively. Leaf-derived callus had the least antioxidant effect. Ascorbic and Gallic acid included in the study as standards had IC50 values of 6.9±0.18 and 8.6±0.65 μg/mL, respectively.Conclusion: An efficient protocol has been established for the induction and proliferation of callus of Rauwolfia vomitoria, justifying the use of tissue culture technique for the conservation of this important medicinal plant.Keywords: Rauwolfia vomitoria, In vitro callus induction, Antioxidant, Plant tissue culture, Conservation

Macroscopic and microscopic evaluation of Triclisia subcordata Oliv.(Menispermaceae) towards its standardization

Background: Triclisia subcordata Oliv. from the family Menispermaceae has been reported for anti-inflammatory, internal wound healing and anticancer activities in ethnomedicine.Objectives: The study was designed to carry out pharmacognostical investigation on the fresh, powdered and anatomical sections of the leaf and petiole of Triclisia subcordata to determine its macro and microscopic characteristics.Materials and Methods: Macroscopic and organoleptic evaluations were carried out on the plant using standard procedures. Microscopic evaluation was done using a light microscope to study the epidermis and transverse sections of the leaf and petiole. Chemo-microscopy of plant sample was done for cell inclusions. The solvent fractions were analysed by the use of thin layer chromatoghraphy for the constituents. Powdered sample and leaf extract were subjected to phytochemical screening and to fluorescence analysis using different organic solvents.Results: Macroscopically, the leaves have a cordate base, an entire margin, caudate apex and a glossy surface. The microscopic evaluation shows the presence of paracytic stomata, unicellular, uniseriated-covering trichomes with an acute apex. The chemo-microscopy revealed the presence of lignin, fats and oils. Phytochemical screening of the powdered leaves revealed the presence of tannins, alkaloids, cardiac glycosides, while flavonoids were absent. The thin layer chromatography showed spots of different retardation factors indicating the presence of different classes of secondary metabolites. The fluorescence analysis also revealed the presence of different colours, which could be employed for the identification of the classes of compounds present in the plant.Conclusion: The results of the study could be useful in setting some diagnostic indices for the identification and preparation of a monograph of the plant.Keywords: Triclisia subcordata, Standardization, Microscopy, Phyto-constituent

Ethnobotanical survey of medicinal plants used as remedy for female infertility and menstrual disorder in southwestern Nigeria

Background: In Sub-Saharan Africa, more than 30% of premenopausal women are affected with secondary infertility. Nigerian (Yoruba) women show a higher tendency towards exaggerated ovarian (PCOS) response to stimulation for assisted conception associated with a higher prevalence of Polycystic Ovary Syndrome.Objective: Identify and document medicinal plants traditionally used to treat female infertility and menstrual disorders among Yoruba ethnic group.Materials and Methods: Data on medicinal plants traditionally used to manage menstrual disorders and female infertility were collected through interviews and Focus group discussions (FGDs).Results: Twenty nine plant species belonging to 20 families were mentioned for management of infertility and menstrual disorder. Plant families mostly used were Euphorbiaceae (20%), Cucurbitaceae (15%), Bignoniaceae (10%), Apocynaceae (10%), Arecaceae (10%) and Solanaceae (10%).Conclusion: This study provides documentation of medicinal plants used in the management and treatment of infertility and menstrual disorder in Southwestern Nigeria.Keywords: Female infertility, Menstrual disorder, Medicinal plants, Bioactive constituent

Providing some pharmacopoeial standards for the leaves of two closely-related Alstonia species

Background: Alstonia boonei De Wild. and Alstonia congensis Engl. (Apocynaceae) are commonly used by the indigenes of Nigeria to treat malaria and other ailments.Objective: This study has examined the leaves of A. boonei and A. congensis with a view to providing some pharmacopoeial standards for identification and development of their individual monograph.Materials and Methods: Macroscopic and microscopic studies were carried out on the leaves of A. boonei and A. congensis with the use of light microscope, while the proximate and fluorescence analyses were carried out according to standard World Health Organization specifications.Results: The leaves of A. boonei and A. congensis were coriaceous. The quantitative microscopy of the leaves showed the upper epidermal cell lengths for A. boonei and A. congensis as 32.5 ± 0.56 (μm) and 38.5 ± 1.8 (μm), while palisade ratios were 5.2 ± 0.4 and 3.7 ± 0.2, respectively. The stomata numbers of A. boonei and A. congensis were 5.0 ± 1.2 and 1.8 ± 0.3, respectively. The stomata length and width of A. boonei, respectively, were 24.5 ± 0.5 (μm) and 22.0 ± 1.2 (μm), while those of A. congensis, respectively, were 35.0 ± 0.8 (μm) and 28.0 ± 3.0 (μm). The vascular bundle in A. boonei was arc-shaped with xylem vessels standing in isolation but assuming a U-shape in A. congensis with xylem vessels interlocking to form a crown-like connection. Vein-islet number of A. boonei (9.0± 0.7) was slightly more than that of A. congensis (8.8±1.8). Total ash values and moisture contents of the leaves of A. boonei and A. congensis were 8.5 ± 0.01 and 6.8 ± 0.03; and 9.1 ± 0.04 and 9.9 ± 0.02, respectively.Conclusion: The foliar epidermal and anatomical characters presented in this study are useful for differentiating between the two closely-related species, Alstonia boonei and Alstonia congensis. The established characters in each of the species could be incorporated into the next editions of the Nigerian and West African Herbal Pharmacopoeias.Keywords: Alstonia spp., Pharmacopoeia, Microscopy, Proximate analysis, Organoleptic evaluatio

Anti – inflammatory and antioxidant effects of Tetrapleura Tetraptera (Schumach & Thonn.) taub. fruit extract in Carrageenan/Kaolin-induced acute monoarthritis in rats

Background: Tetrapleura tetraptera has been reported traditionally for the treatments of various ailments, which include convulsion, leprosy, inflammation and rheumatic pains, schistosomiasis, asthma and hypertension. The antiinflammatory activity of the aqueous extract of T. tetraptera fruit in egg albumin-induced paw oedema as well as in vitro anti-inflammatory and antioxidant activity had been documented. However, there had been no studies on the antiarthritic properties of the plant.Objectives: The present study was designed to investigate in vivo anti-inflammatory and anti-oxidant activities of the methanol extract Tetrapleura tetraptera in carrageenan/kaolin-induced acute monoarthritis in rat.Materials and methods: The acute toxicity of the methanol plant extract was determined using Lorke’s method. In vivo carrageenan/kaolin induced arthritis assay was performed on Tetrapleura tetraptera (TtME) extract (100, 200 and 400 mg/kg) using albino rats. Indomethacin (10 mg/kg) and normal saline were employed as positive and negative controls, respectively. Histological study was performed on the knee joint of the animals after collection of blood for antioxidants analysis.Results: Acute toxicity study showed that TtME is safe by oral administration up to 5000 mg/kg body weight. Oral administration of TtME and indomethacin (10 mg/kg) produced a significant (p < 0.05) time and dose-dependent decrease in joint diameter. There was a decrease in malondialdehyde and increase in glutathione and superoxide dismutase. The histological assessment revealed no major adverse effects on the animals.Conclusion: Tetrapleura tetraptera fruit extract possesses anti-arthritic activity, which may be attributed to its antiinflammatory and antioxidant activity.Keywords: Tetrapleura tetraptera, Antioxidant, Anti-inflammatory, Anti-arthritic activity, Histopatholog

Quantification of endogenous phytochemicals and determination of their exogenous effects in somatic embryogenesis pathways of white and water yams

Open Access ArticleIn vitro propagation of yam via organogenesis is constrained with low multiplication rate. Somatic embryogenesis (SE) has shown rapid multiplication potentials in yam. However, it has not been adopted by practical seed system scenarios due to genotype specificity. Reports have shown that SE is regulated endogenously by phytochemicals, but this is yet to be elucidated for yam. This study identified, quantified endogenous, and evaluated effects of exogenous application of selected identified phytochemicals in yam SE. Callus was induced from in vitro axillary bud explants of three Dioscorea rotundata genotypes in Murashige and Skoog (MS) medium containing 9.1 µM 2,4-dichlorophenoxyacetic acid and 5.4 µM naphthaleneacetic acid. Plantlets were regenerated using MS medium containing 4.4 µM benzylaminopurine and 34.0 µM uniconazole-P. Endogenous phytochemicals associated with axillary bud, calluses, and plantlets were identified and quantified using GC/MS. Effect of selected identified phytochemicals on the genotypes was investigated in a 5 × 6 factorial in completely randomized design (r = 3). Data taken on plantlet regeneration was analyzed using ANOVA at α0.05. A total of 27, 22, and 35 phytochemicals were identified in Kpamyo, Ekiti2a, and Asiedu, respectively. Hexamethylcyclotrisiloxane (36.4%, Kpamyo), Tris-tert-butyldimethylsilyloxy-arsane (59.3%, Ekiti2a), and 4-methyl-2-trimethylsililoxy-acetophenone (52.7%, Asiedu) were highest in callus. N-Methyl-1-adamantaneacetamide (31.8%, Kpamyo) and Tris-tert-butyldimethylsilyloxy-arsane (52.7%, Ekiti2a, Asiedu) were highest in plantlets while Tris-tert-butyldimethylsilyloxy-arsane (41.2%, Kpamyo), hexamethylcyclotrisiloxane (55.8%, Ekiti2a), and erythro-9,10-dibromopentacosane (38.9%, Asiedu) were highest in axillary bud. Plantlet regeneration differed significantly among phytochemicals and ranged from 0.7 ± 0.3 (control) to 4.5 ± 0.5 (40.5 µM phenylacetic acid). Also, genotype × phytochemical interactions on number of plantlets regenerated were significant, and mean values ranged from 0.0 ± 0.0 (TDa2014, 4.8 µM decamethyltetrasiloxane) to 7.0 ± 1.7 (TDa2014, 40.5 µM phenylacetic acid). The application of 40.5 µM phenylacetic acid enhanced plantlet regeneration in Kpamyo and TDa2014 by 5.39% and 343.04%, respectively

Exploitation of petiole, nodal segment, bulbil and tuber anatomy for species identification in Dioscorea Linn. species from Oyo and Ekiti states- southwestern Nigeria

Herb sellers and Traditional Medical Practitioners often substitute D. hirtiflora, cultivars of D. bulbifera and D. dumetorum with closely related cultivars within species during herbal preparations. This practice, which may be as a result of mis-identification, may be very injurious to users of herbal products. The present study investigated anatomical characters of three wild and edible species of Dioscorea used in ethnomedicine in Southwestern Nigeria with a view to aiding species identification. Thin sections of the median petiole and nodal segments were prepared using free hand sectioning. Bulbils and tubers stored in 50% ethanol were sectioned at 5 μm with the aid of a sledge microtome. Sections were stained in Safranin O solution for 2-10 min while sections from bulbils of D. hirtiflora were counter stained with toluidine for 1 min and mounted with glycerol. All slides were examined under the light microscope at x100 and x400 magnifications and photos were taken using digital camera mounted on Zenith Ultra-500 A light microscope. Petiole and nodal segments anatomy showed six and nine vascular bundles, respectively in D. hirtiflora, whereas eight and eleven bundles were observed in D. dumetorum and D. bulbifera. Petioles of D. hirtiflora and D. dumetorum were profusely surrounded with stellate and simple unicellular trichomes. Parenchyma cells in wild D. dumetorum were beaded, while they were not in the edible cultivar. Generally, scattered vascular bundles, layers of collenchymas cells, and dilated parenchyma cells filled with tannins were observed in the bulbils/tubers of all species. Anatomical characters in the selected Dioscorea species are taxonomically significant for species identification and could serve as diagnostic taxonomic tools for their standardization.Keywords: Anatomy, Dioscorea spp., Identification, Medicinal applicatio

Ethnobotanical survey of tree species used for wound healing in Ibadan, southwest Nigeria

The disruption of cells and tissues at the sites of injury leads to varying degrees of wounds, which can be as a result of the physical, chemical, microbiological or immunological process. The present study reports the findings from an ethnobotanical survey of three Local Government areas (LGAs) in Ibadan, Oyo State in Southwest Nigeria, carried out to document tree species and plant parts used in the treatment of wound and related skin disorders. Semi-structured questionnaires were used to interview the herb sellers, traditional medical practitioners, and some elders in the study area comprising of Ibadan North, Ibadan Southeast and Ibadan Northwest LGAs. Eighty-five respondents were questioned on their knowledge of skin diseases and wound care management. Seventy-one plants out of which sixty-five were tree species belonging to thirty angiosperm families were identified as plant species used for the treatment of wound and related skin disorders. Some of the most frequently used tree species mentioned by the respondents are: Khaya grandifoliola C. D. C., Vitellaria paradoxa C.F. Gaertn., Xylopia aethiopica (Dunal) A. Rich, Kigelia africana (Lam.) Benth., Bombax buonopozense P. Beauv., Terminalia glauscescens Planch. ex Benth. and Lophira alata Banks ex Gaertn. The most frequently mentioned family is Fabaceae, followed by Meliaceae and Annonaceae. Stem barks were predominant in the prescription. Preparations including two or more plant species were common. However, in some cases, prescriptions were in powder form. The mode of preparation and dosages are presented in this paper. The results of this study revealed many tree species used to treat wounds and skin disorders in the three Local Government Areas surveyed in Ibadan, Southwest Nigeria.Keywords: Wound healing, ethnobotanical survey, tree species, medicinal uses, conservatio

Chemical detection of cysteine-rich circular petides in selected tropical Violaceae and Moringaceae families using modified G-250 and mass spectrometry

Cysteine-rich circular peptides (CRCs) comprise a large family of gene encoded and low molecular weight polypeptides that has recently engaged the attention of scientists. This class of peptides exhibit a continuous circular configuration and a cystine knot backbone, which defines their resilient nature-directed structural design. Many CRCs have been reported in medicinal plants the first of which is kalata B1 cyclotide from the traditional African plant Oldenlandia affinis. Their detection and isolation can be very challenging and evasive. Only about 1% of plant species have so far been reportedly screened. A modified preliminary chemo-microscopic/macroscopic method involving the use of G-250 stain was applied followed by thin layer chromatographic protosite reaction for plant selection. This was followed by the Matrix Assisted Laser Desorption Time of Flight (MALDI-TOF) Mass spectrometry guided experiment for cyclotide discovery. The blue colouration produced upon sample reaction with modified G-250 led to the selection of potential circular peptide containing plant samples. A further MALDI-TOF MS-guided screening resulted in the detection of circular peptides and cyclotides in Moringa oleifera, Rinorea dentata, R. oblongifolia and R. brachypatela. Viola odorata and Viola tricolor (positive controls) indicated the presence of cyclotides. Results from this study can serve as proof-of-concept for plant selection based on preliminary cysteine-rich circular peptide detection in plants especially with the use of G-250 stain.Keywords: Cysteine-rich Cyclotides, Violaceae, Moringaceae, TLC, Microscopy, MALDI-TOF M

Particulate matters pollution in selected areas of Nigeria: Spatial analysis and risk assessment

The increasing rate of unregulated emissions of particulate matters from different sources in Nigeria is of great concern. This study therefore focused on monitoring particulate matters (PM2.5 and PM10) level in some municipal areas in Nigeria. The study also simulated the dispersion pattern of the particulate matters and assessed the health risk of particulate matter exposure. PM2.5 and PM10 level in the selected areas were quantified for 8 months using Centre for Atmospheric Research (CAR) satellite-based sensors. Hybrid single-particle Lagrangian integrated trajectory (HYSPLIT) model was used to simulate the dispersion pattern and the dispered particulate matter position. The risk assesment was estimated by air quality index (AQI) using World Health Organization (WHO) guideline for PM2.5 and PM10 as the standard. The result showed that the levels of PM2.5 and PM10 on monthly average basis for the selected areas were all above the WHO guideline. The dispersion model and potential particulate matter position revealed that it takes almost 12 hours for any released particulate matter to be fully dispersed and that majority of the dispersed particles were found within 100 m. The AQI level of PM2.5 ranged from 65.13 to 927.07 while PM10 AQI ranged from 73.51 to 256.65 for the selected regions, which showed poor and unhealthy air quality. Also, the inhabitants of the studied areas are at high risk of particulate matters exposure which may lead to a reduced life expectancy over a long period of exposure. This study however concluded that a more robust measure should be put in place to ensure an improved air quality