Ethnoveterinary medicinal plants and their utilization by the people of Soro District, Hadiya Zone, southern Ethiopia.

BACKGROUND: Ethnoveterinary studies are important to maintain the sustainability of livestock health and support people's livelihoods through the provision of food, maintaining livestock health, and other biological resources. This study was carried out in Soro District, southern Ethiopia, to identify, document and analyse plant species with ethnoveterinary uses along with the associated indigenous and local knowledge. MATERIALS AND METHODS: Informants were selected using purposive (key informants) and systematic random sampling (general informants) methods. Data on ethnoveterinary plants and their uses were collected through semi-structured interviews, guided field walks, 13 focus group discussions with five to seven members in each and participant observation. Informant consensus factor and fidelity level were computed to identify the most common livestock ailment categories and the best plant species with ethnoveterinary use, respectively. Preference ranking methods were used to identify the potentially effective ethnoveterinary medicinal plants for the most frequently reported livestock ailments. The use diversity of multipurpose plants with ethnoveterinary importance was analysed using the analytical methods of ethnobotany including priority ranking, comparisons and important indices. The T-test statistic was used to compare knowledge differences among different social groups. RESULTS: A total of 132 plant species in 120 genera and 61 families were reported by informants as having ethnoveterinary uses. The plants are said to be used by the local communities in various ways to treat 50 livestock health problems. Higher number of informants (23.77%) cited Momordica foetida for the treatment of 16 livestock ailments. The highest informant consensus value for this species is associated with its use for treating blackleg in cattle; Nicotiana tabacum was cited for the treatment of 15 livestock ailments mainly recommended for the Lumpy Skin Disease/Ailment of bovines; Croton macrostachyus for treatment of 13 livestock ailments including wooden tongue, FMD in bovines; and Gymnanthemum amygdalinum for nine ailments mainly diarrhoea of all livestock types. Achyranthes aspera is claimed to provide the most effective treatment for Aspiration pneumonia (severe coughing in bovines, sheep and goats) alone, while Croton macrostachyus, Ximenia americana, Allium sativum and Juniperus procera were indicated as potential plant species to treat Lumpy Skin Disease in bovines in the order given. The fidelity level analysis showed that Datura stramonium, Dodonaea viscosa subsp. angustifolia and Asparagus africanus were potential medicinal plant species to treat the respective ailments of rabies, Peste des petits ruminants (PPR) and evil eye/spirit. Multipurpose plant species including Prunus africanus, Combretum molle and Afrocarpus falcatus have been highly threatened as indicated by direct matrix ranking mainly due to collection of fuel wood, construction materials and making household utensils, and farm implements rather than for other uses. CONCLUSION: Soro District has rich and diversified livestock herbal medicinal resources, and indigenous knowledge of remedy preparations and applications is transmitted through generation lines. This resource faces anthropogenic threats with deforestation being the leading factor. Consequently, ethnoveterinary medicinal plants continue to decline before adequate and proper scientific documentation and testing are made. There is a dire need for planning and implementation of appropriate in situ and ex situ conservation strategies and to strive towards ensuring the survival and sustainable utilization of such important plant resources of Soro District. This must be supported by further documentation of the associated indigenous knowledge and pharmacological testing of the key promising species including Balanites aegyptiaca (novel species/NS to treat specific ailment), Brugmansia suaveolens (novel species/NS reported first to treat Livestock ailments/LsAs), Euclea divinorum (NS to treat specific ailments), Grevillea robusta (NS), Hagenia abyssinica (NS for the reported specific ailment), Pentanema confertiflorum (NS), Juniperus procera (NS), Maesa lanceolata (NS), Millettia ferruginea (NS for reported specific ailments), Schrebera alata/NS, Securidaca longepedunculata, Spiniluma oxyacantha/NS, Vepris nobilis (novel species reported first to treat LsAs), Zanthoxylum asiaticum /NS and Ximenia americana (NS for specific ailments). This ethnoveterinary study attempted to fill part of the gaps concerning the prevalent livestock health problems and the associated indigenous and local knowledge in the area.

In vitro antimicrobial and antioxidant activities, essential oil composition, and in silico molecular modeling analysis of secondary metabolites from roots of Verbascum sinaiticum.

Verbascum sinaiticum is locally used to treat wound, stomachache, viral infection, cancer, sunstroke fever, abdominal colic, diarrhea, hemorrhage, anthrax, and hepatitis. The objective of this study was to identify the compounds and to evaluate the antimicrobial and antioxidant activity of the extracts and isolated compounds from V. sinaiticum. The 1H-NMR, 13C-NMR, and DEPT-135 were used to elucidate the structures of isolated compounds. Essential oils were extracted by hydrodistillation method and their chemical analyses were performed by GC-MS. The broth microdilution method was used to evaluate the antimicrobial activity. The radical scavenging activity of the extracts and isolated compounds were evaluated using DPPH method. Silica gel column chromatographic separation of root extracts afforded seven known compounds: 3'-(4''-methoxy phenyl)-3'-oxo-propionyl hexadecanoate (1), harpagoside (2), pulverulentoside I (3), scrophuloside B4 (4), scropolioside A (5), scropolioside-D2 (6), and harpagide 6-O-ß-glucoside (7), which are all reported from this species for the first time. The EO extracts from leaves and roots were the most susceptible to Streptococcus agalactiae, with a 2 mg/mL MIC. The EO from roots was effective against Candida albicans and Trichophyton mentagrophytes, with a MIC of 8 mg/mL. The MeOH and CH2Cl2/CH3OH (1:1) root extracts showed the maximum activity against S. epidermidis with MIC values of 0.25 mg/mL. The strongest antibacterial effects were demonstrated against Staphylococcus epidermidis, which exhibited a 0.0625 mg/mL MIC for compound 1. The strongest radical scavenging activity was exhibited by the methanol extract (IC50 = 3.4 µg/mL), and compounds 4, 6, 5, 3, 7, and 2 with IC50 values of 3.2, 3.38, 3.6, 3.8, 4.2, and 4.7 µg/mL, respectively, in comparison with ascorbic acid (IC50 = 1.3 µg/mL). The results of the molecular docking analysis of compounds revealed minimal binding energies range from -38.5 to -43.1 kJ/mol, -33.1 to -42.7 kJ/mol, -34.7 to -39.3.7 kJ/mol, -25.5 to -37.6 kJ/mol against human myeloperoxidase (PDB ID: 1DNU), murA enzyme (PDB ID: 1UAE), human topoisomerase IIß (PDB ID: 4fm9), S. epidermidis FtsZ (PDB number: 4M8I) proteins, respectively. The docking results and the in vitro antibacterial activity are in good agreement. These findings show that the isolated compounds 2-7 can act as potential antioxidants and strong antibacterials against Staphylococcus aureus and S. epidermidis. As a result, V. sinaiticum root extracts have the potential to be effective in treating diseases caused by bacteria and free radicals, as long as further investigation has been suggested for the ultimate decision of this plant's potential candidate.

Antibacterial and phytochemical analysis of traditional medicinal plants: An alternative therapeutic Approach to conventional antibiotics.

The purpose of this study was to carry out antibacterial and phytochemical analyses on six selected medicinal plants that have been traditionally used by the local people to treat and control different diseases. The antibacterial activities of methanolic extracts of these plants were assessed using the Agar well diffusion and Microtiter broth dilution methods. The root extract of Andrachne aspera showed significantly (p < 0.05) highest mean zone of inhibition at concentrations of 100 mg/ml (33 ± 0.17) and 200 mg/ml (33.5 ± 0.84) against S. epidermidis. The second highest mean zone of inhibition (24.8 ± 0.41) was recorded by Dichrostachys cinerea leaf extract against S. epidermidis at 200 mg/ml concentration. The minimum inhibitory concentrations 1.0 ± 0.0 was recorded by Andrachne aspera against E. faecalis and 2.0 ± 0.0 against S. aureus by Dichrostachys cinerea. The preliminary phytochemical analysis showed that Andrachne aspera and Dichrostachys cinerea contained strong concentration of Polyphenols and Flavonoids. Therefore, these two medicinal plant species have promising potential for further detailed investigations, including safety tests, characterization and isolation of bioactive secondary metabolites for the development of alternative drugs.

Evaluation of Anti-Inflammatory Activity of the Methanol Extracts of Premna schimperi Engl (Lamiaceae) Leaves in Rats.

Background: Even though it is a protective reaction, inflammation continues to be one of the most challenging medical disorders. The current conventional anti-inflammatory drugs have many undesirable health effects and are in need of newer drugs. The purpose of this study was to evaluate the anti-inflammatory effects of an aqueous methanol crude extract of Premna schimperi leaves. Methods: Premna schimperi leaf was extracted with 80% methanol and concentrated; the concentrated extract was used to evaluate the acute toxicity and anti-inflammatory effects. For the acute toxicity study, a single dose of Premna schimperi extract at a dose of 2000 mg/kg was administered and observed for 14 days. Acute, sub-acute, and chronic anti-inflammatory models were employed to evaluate the anti-inflammatory effect of the extract compared to the standard drug. Data were analyzed with SPSS V. 27, and the significance was established with a one-way ANOVA followed by a post hoc Tukey's test. Results: Acute oral toxicity testing at a dose of 2000 mg/kg did not show any sign of toxicity. According to the phytochemical study, the plants contained flavonoids, terpenoids, tannins, cardiac glycosides, steroids, phenolics, and anthraquinones. The extract doses of 200 mg/kg, 400 mg/kg, and 800 mg/kg of extracts effectively (p<0.001) reduced paw edema in the acute and sub-acute models of inflammation. When compared to the negative control group, all tested doses in the chronic model significantly (p<0.05) decreased the production of exudates and the amount of granuloma tissue. Conclusion: Premna schimperi displayed significant anti-inflammatory activity. The tested doses inhibit the formation of edema, granulomas, and exudates.

Antidiarrheal Activities of the Methanol Leaf Extracts of Olinia rochetiana (Oliniaceae) Against Castor Oil-Induced Diarrhea in Mice.

Background: Olinia rochetiana has been used traditionally to cure diarrheal disease. Therefore, this study aimed to investigate the acute toxicity and antidiarrheal effect of O. rochetiana leaf extracts. Methods: Cold maceration was used to extract plant leaf powder with 80% methanol. The extract's antidiarrheal action was tested against a castor oil-induced diarrheal model, a charcoal meal test, and enteropooling tests at doses of 100, 200, and 400 mg/kg. Negative controls received the vehicle at 10 mL/kg, while positive controls received loperamide at 3 mg/kg. Results: From the study, no apparent toxicity was observed when a single dose of 2000 mg/kg was administered. In the castor oil-induced model, the extract delayed the onset of diarrhea, reduced stool frequency, and decreased wet feces weight and number in a dose-dependent manner at 200 mg/kg (p < 0.05) and 400 mg/kg (p < 0.01). The percent reduction in moist feces at 100, 200, and 400 mg/kg was 54.2, 23.97, and 18.26%, respectively, indicating a significant dose-dependent decrease. In a charcoal meal test, the extracts at 200 and 400 mg/kg revealed a peristaltic index of 65 and 46%, respectively, with considerable inhibition of charcoal transport at 23 and 39%. The weight and volume of intestinal contents dropped significantly at a dose of 400 mg/kg (p < 0.01), which is 0.43 mg/kg, in the enteropooling test when compared with the tested dose. The computed in vivo antidiarrheal index revealed diarrheal inhibition values of 46.06 and 71.06% at 200 and 400 mg/kg, respectively. Conclusion: In the current investigation, O. rochetiana showed significant antidiarrheal activity with no symptoms of toxicity in mice.

Acute and developmental toxicity of embelin isolated from Embelia schimperi Vatke fruit: In vivo and in silico studies.

Background: Embelin is a hydroxybenzoquinone constituent of the Embelia species that has anti-disease properties. However, its toxicity, particularly the in silico, acute, and developmental toxicity profiles, has yet to be thoroughly investigated. Hence, this study aims to assess these toxicity profiles. Materials and Methods: In silico and in vivo experimental studies were conducted on embelin isolated from the fruits of Embelia schimperi Vatke. In silico toxicity predictions were computed using the ProTox model. The in vivo experiment was done by administering 5000 mg/kg of embelin to a single female albino Wistar rat, followed by three female rats in the absence of death, to determine the mean lethal dose (LD50). Afterwards, three groups of pregnant rats were treated with embelin at doses of 250 mg/kg, 500 mg/kg, and 1000 mg/kg for the developmental toxicity test. Vehicle and ad libitum control groups were used to compare the acute and developmental toxicity variables. Results: In silico toxicity predicted that embelin is free from hepatotoxic, carcinogenic, mutagenic, and cytotoxic effects. No inhibitory effect on hERG channels was observed. It has an immunotoxic property and an inhibitory effect on the CYP2D6 enzyme. Since mortality and signs of toxicities were not observed after treatment with 5000 mg/kg, the mean lethal dose (LD50) is determined to be > 5000 mg/kg. There was no significant difference in the morphological scores or number of somites among experimental animals. None of the embryonic systems possessed developmental delays. Nevertheless, the crown-rump length of the high-dose group became significantly shorter. Maternal food intake and weight gain exhibited significant dose-dependent differences between embelin-treated animals and controls. The number of implantations was significantly low in the treatment group, accompanied by a higher frequency of prior resorption. Conclusion: Embelin is predicted to have a high probability of immunotoxicity potential and affect drug metabolism by inhibiting CYP2D6. In addition, it affects food intake, weight gain, and the number of implantations in pregnant rats. Therefore, it is highly recommended not to take embelin and embelin-rich plants during pregnancy. Further in vitro and in vivo studies need to be conducted to understand the mechanism behind the toxicity of embelin.

Analgesic and Anti-Inflammatory Activities of 80% Methanol Extract and Solvent Fractions of Ehretia cymosa Thonn (Boraginaceae) Leaves in Rodents.

Background: Ethnobotanical studies in various districts of Ethiopia reported that Ehretia cymosa (E. cymosa) is used for the management of headache, abdominal pain, arthritis and rheumatism. However, there is no scientific investigation done so far to confirm these traditional claims. Thus, the aim of this study was to assess the analgesic and anti-inflammatory effects of the 80% methanol extract and fractions of E. cymosa leaves. Methods: The dried and pulverized leaves of E. cymosa were soaked with 80% methanol to obtain a crude extract. Fractionation was done using chloroform, ethyl acetate and water by a soxhlet apparatus. The analgesic effects of the crude extract and solvent fractions were assessed using acetic acid-induced writhing and hot plate tests whereas anti-inflammatory activities were investigated using carrageenan-induced paw edema and cotton-pellet-induced granuloma models. Results: In all the tested doses, the 80% methanol extract and solvent fractions revealed substantial (p < 0.001) analgesic activities in acetic acid induced writhing test. In the hot plate method, all the tested doses of E. cymosa crude extract and the solvent fractions produced significant analgesic activities (p < 0.05). In the carrageenan-induced acute inflammation model, all tested doses of the crude extract and solvent fractions resulted in a significant decline in paw edema. The 80% methanol extract and solvent fractions of E. cymosa at all the tested doses significantly reduced inflammatory exudates and granuloma mass formations (p < 0.001). Conclusion: From the results of this investigation, it can be stated that 80% methanol extract, aqueous, ethyl acetate and chloroform fractions of E. cymosa exhibited considerable analgesic and anti-inflammatory activities, supporting the plant's traditional use as a remedy for a variety of painful and inflammatory conditions.

Composition of the Essential Oil Thymus schimperi and Evaluation of Its Acute and Subacute Toxicity in Wistar Albino Rats: In Silico Toxicity Studies.

BACKGROUND: In Ethiopian traditional medicine, the aerial part of Thymus schimperi is widely used to treat diseases such as gonorrhea, cough, liver disease, kidney disease, hypertension, stomach pain, and fungal skin infections. However, there is insufficient investigation on the toxic effect of the essential oil of T. schimperi. The aim of this study was, therefore, to evaluate the acute, subacute, and in silico toxicity of Thymus schimperi essential oil in the Wistar albino rats. METHOD: Essential oil of the aerial part of T. schimperi extracted by hydrodistillation was analyzed by GC-MS. The oil was subjected to toxicity studies. In the acute toxicity study, rats were randomly divided into seven groups (n = 5). The control group received only distilled water with 2% of tween 80, whereas the experimental groups received single doses of 300, 600, 900, 1200, 1500, and 2 000 mg/kg of the oil. In the subacute toxicity study, rats were randomly divided into four groups (n = 10). The control group received distilled water with 2% of tween 80, whereas the experimental groups received 65 mg/kg, 130 mg/kg, and 260 mg/kg of the oil orally for 28 days. At the end of the experiment, blood samples were collected for hematology and clinical chemistry evaluation. Gross pathology and histopathology of the liver and the kidneys were also evaluated. For the in silico toxicity study, PubChem CID numbers of GC-MS identified bioactive compounds in the essential oil of T. schimperi obtained from PubChem. Chemdraw (8.0) was used to construct two-dimensional structures of the compounds. The Swiss ADMET web tool was used to convert the two-dimensional structures into a simplified molecular-input line input system (SMILES). In addition, the toxicity parameters were predicted via vNN and ADMET servers. RESULTS: In this study, the LD50 of the essential oil of T. schimperi was found to be 1284.2 mg/kg. According to the World Health Organization, the oil is classified as moderately hazardous in its oral administration. In the subacute toxicity study, rats showed no significant changes in behavioral indices, gross pathology, body weight, biochemical, and most hematological parameters. However, hematological profiles showed a significant decrement in WBC counts and a significant increment of MCV in high dose (260 mg/kg) groups as compared to the control group. Furthermore, no significant differences were observed between the control and essential oil-treated groups, observed in the gross histopathology of the liver and the kidneys. In the in silico toxicity study, all compounds derived from the essential oil showed no cardiac toxicity (h-ERG Blocker), AMES (Ames Mutagenicity), and cytotoxicity via ADMET and vNN-ADMET toxicity predictors. However, by using these servers, about 8.6% of the compounds showed hepatotoxicity, only 3.45% caused drug-induced liver injury, and only 1.75% were potentially toxic to the mitochondrial membrane. CONCLUSION: From the results of this study, oral administration of the essential oil T. schimperi up to a dose of 130 mg/kg is not harmful. However, in the high-dose (260 mg/kg) group, the WBC count was significantly decreased and the MCV was significantly increased. In the in silico toxicity study, most of the components of the oil were found to be nontoxic, although a few of the compounds showed hepatotoxicity and mitochondrial membrane potential toxicity. It is, therefore, essential to conduct chronic toxicity of the essential oil as well as its components, which showed toxicity in the in silico study before using preparations containing the essential oil of T. schimperi.

Development, Characterization, and Evaluation of Novel Broad-Spectrum Antimicrobial Topical Formulations from Cymbopogon martini (Roxb.) W. Watson Essential Oil.

BACKGROUND: Skin infections were the most frequently encountered of all infections and the 4th leading cause of nonfatal disease burden. Topical drugs have been used for the management of skin infections. The growing concern of drug resistance to the topical agents has warned the need for continuous development of novel drug. Essential oils are the best candidate for new drug with different mode of action and target as they are rich in chemical constituents. OBJECTIVE: To evaluate and develop safe and effective topical antimicrobial formulations from essential oil of Cymbopogon martini. Method. Essential oil was extracted using hydrodistillation aerial part C. martini and topical formulations were prepared in five different semisolid bases. In vitro antimicrobial investigations were performed on essential oil and topical formulations. Skin sensitizations of the formulations were evaluated using guinea pig maximization. RESULTS: The essential oil of C. martini has shown broad-spectrum antimicrobial potency against all tested organisms with MIC value ranging from 0.65 to 10 µg/ml. Absolute inhibitions of growth of fungi were observed against Trichophyton mentagrophytes and Trichophyton rubrum at concentrations above 1% of oil and against Microsporum canis and Trichophyton verrucosum at a concentration of 4% oil. Among topical formulations, the highest antimicrobial activity was recorded in hydrophilic ointment followed by macrogol blend ointment. The antimicrobial activity of oil was higher in fungal pathogen compared to bacteria. Gram positive bacteria were more sensitive than gram negative bacteria. Hydrophilic and macrogol blend ointment containing 5% oil did not produce any skin sensitization on guinea pigs. CONCLUSION: In conclusion, topical formulations of C. martini essential oil can be alternative topical agents with safe broad-spectrum activity for the treatment of skin disorder. Further studies should focus on shelf life study and clinical study of the product.