In Vitro Biocompatibility Assessment of Entomopathogenic Fungi and Plant Leaf Extracts

Maize (Zea mays L.) is one of the important and basic cereal crops in the world. Fusarium verticillioides is one of the most important pathogens causing root, stalk, ear rot, and seedling blight diseases in maize. In recent years, the restriction of the use of most chemical fungicides has led to the importance of research on plant resistance inducers. Different parts of the Melia azedarach L., such as bark, leaf, seeds, and root have very strong medicinal value, and the seeds of this species can also be used in the production of fungicides and pesticides in agricultural products. In this study investigated the effects of Melia azedarach leaf and fruit extracts on Kale (F1) corn seeds infected with Fusarium verticillioides. Six different applications (Positive Control, Negative Control, Fruit Extract, Leaf Extract, Fruit Extract + GA3, Leaf Extract + GA3) were established in a randomized plot design with three replications. According to the results, the highest value for germination rate on the 3rd day was 67.78% in the negative control group, and the highest value for germination rate on the 7th day was obtained in the positive control 77.77% and negative control 74.44%. In terms of Fusarium contamination rates, the highest values on the 3rd and 7th days were found to be 36.67% and 50.00% in the fruit extract group, respectively. Root length was determined as the highest value at 47.03 mm in the leaf extract group and the lowest value at 16.00 mm in the fruit extract + GA3 group. In shoot length, leaf extract gave the best results at 57.13 mm and leaf extract + GA3 as 49.23 mm. The findings show that Melia azedarach extracts have positive effects on germination and growth in Fusarium-infected seeds. It is thought that this natural solution may provide positive contributions to maize cultivation.

Medicinal plants have been used for the maintenance of health for many years. Emerging cases of adverse drug reactions, high cost, and antimicrobial resistance using a single antimicrobial drug have heightened the need for research into plant sources of antimicrobial activity like Melia azedarach and Solanum incanum. This research investigated the synergistic antimicrobial activity exhibited by ethanolic leaf extracts obtained from Melia azedarach and Solanum incanum when tested against the pathogenic bacterium Escherichia coli. The study aimed to characterize the presence and composition of phytochemicals in the individual extracts, thereby elucidating their contribution to the observed antimicrobial effects. Additionally, minimum inhibitory concentration (MIC) assays and disc diffusion tests were performed to determine the optimal concentration required to inhibit the growth of Escherichia coli. Solanum incanum at a concentration of 2g/ml showed an average zone of inhibition of 6.92mm while Melia azedarach exhibited a zone of inhibition of 6.97 mm for Escherichia coli. The combined extracts showed a zone of inhibition of 8.34 mm for Escherichia coli. The minimum inhibitory concentration for Solanum incanum was 0.5g/ml while that of Melia azedarach was 1.4g/ml for Escherichia coli. Leaf extracts of both Melia azedarach and Solanum incanum had moderate to high levels of tannins, coumarins, flavonoids, and anthraquinones. Alkaloids and saponins were in low amounts in both extracts while steroidal glycosides were in moderate amounts in Melia azedarach but high in Solanum incanum. In conclusion, both plant extracts demonstrated significant antibacterial effects with both plants having a similar potency in terms of zone of inhibition diameter. The combined leaf extract of both plants had higher activity compared to individual plant extracts. We thus recommend that the combined extract can be used as an alternative modality for treating E. coli infections of mild severity without complications.

Biological activities of Boswellia sacra extracts on the growth and aflatoxins secretion of two aflatoxigenic species of Aspergillus species

Abstract. Njogu MJ, Murungi JI, Wanjau RN. 2021. Effects of mineral levels and leaf extracts of some plants on soil pH and growth rate of selected non-leguminous plants. Cell Biol Dev 5: 76-89. The purpose of this study was to determine the effect of growth rate on non-leguminous plants such as wheat (Triticum aestivum L.), kale (Brassica oleracea var acephala), and coriander (Coriandrum sativum L.) grown on acidic soils treated with leaves and leaf extracts of plants known to have a high buffering capacity and mineralization capacity, i.e., Jacaranda mimosifolia D. Don, Cordia africana Lam, Croton macrostachyus Hochst. ex Delile, Vitex keniensis Turril, Tithonia diversifolia (Hemsl.) A.Gray, Manihot esculenta Crantz, Carica papaya L., and Helianthus annuus L. For this investigation, the leaves and leaf extracts were applied to the soil with a pH of 4.75. Leaf extracts and leaves of eight species were combined according to the macronutrient content and mineralization of chosen plants discovered in Nyandarua County, Kenya. Wheat, kale, and coriander growth rates were determined by comparing the dry mass of uprooted seedlings from the trial and control studies every fourteen days for 60 days. Flame photometry, atomic absorption spectrometry (AAS), UV/visible spectroscopy, and turbidimetry were used to determine macronutrients in leaf extracts. The analysis of variance (ANOVA) and Student-Newman-Keuls (SNK) tests were used to analyze the data. For 60 days, the mean pH value of leaf extracts ranged from 4.580.27a to 7.440.04d. K+; 189.95±0.17c, Ca2+; 367.33±0.67e, Mg2+; 114.33e±0.33, PO43-; 55.38±0.23f, NO3-; 322.25±0.40f, and SO42-; 56.48±0.23e had the greatest mean levels in leaves (?g/g). Macronutrient levels were significantly different in soil treated with leaves and leaf extract (Le), commercial NPK fertilizers (Cf), and untreated soil (Us). Wheat (dry mass) grew at a mean rate of 1.27±0.13b g/wk (Le), 1.26±0.12b g/wk (Cf) and 0.32±0.02a g/wk (Us), while in (height) was 7.29±1.43b cm/wk (Le), 6.20±1.95b cm/wk (Cf), 3.98±0.97a cm/wk (Us). For kales 0.20±0.01a g/wk (Le), 0.30±0.02a g/wk (Cf), and 0.03±0.01b g/k (Us), while in (height) 2.15±0.85c cm/wk (Le), 2.57±0.88b cm/wk (Cf), and 1.04±0.02a cm/wk (Us). The mean growth rate for corianders was 0.16±0.01a g/wk (Le), 0.17±0.02a g/wk (Cf), and 0.10±0.01a g/k (Us), while in (height) 1.85±0.56b cm/wk (Le), 1.86±0.58b cm/wk (Cf) and 0.79±0.18a cm/wk (Us). This study's findings show that the plant's leaves and leaf extracts can be used in place of inorganic fertilizers and should be recommended for agroforestry in low-pH environments.

Fusarium oxysporum has been the enemy in agriculture due to the wide range of infections in the whole crop and postharvest. Botanical extracts in traditional medicine systems were considered valuable sources for discovering new antifungals. Thus, exploration to get antifungals from eco-friendly botanical extracts as an alternative to synthetic fungicides needs to be expanded. Therefore, this study aims to test the antifungal potential of seven eco-friendly botanical aquadest extracts against F. oxysporum in vitro. The antifungal potential test was done on PDA media and the mycelial growth data was collected every two days for seven days. There were three antifungal phenomena were observed. The pandan leaf extract had a 4% fungal inhibition zone (phenomenon I). The extract of banana leaf, Hibiscus flower, papaya leaf, guava twig (phenomenon II), orange peels, and betel leaf (phenomenon III) had no values of fungal inhibition zone. Thus, pandan leaves extract revealed a fungal inhibition while other six botanical extracts had not enough antifungal potential in vitro.

The efficacy of leaf extracts of basil (Ocimum basilicum), bitter leaf (Vernonia amygdalina), lemon grass (Cymbopogen citratus), neem (Azadirachta indica) and paw-paw (Carica papaya) on major seed-bornefungi: Aspergillus niger, Aspergillus flavus, Botryodiplodia theobromae and Fusarium moniliforme of African yam bean ( Sphenostylis stenocarpa ) seeds, and on seed germination and seedling emergence were studied in vitro and in vivo. All the plants leaf extracts (crude and aqueous) significantly (P . 0.05) reduced the incidence of seed-borne fungi tested and increased seed germination and seedling emergence except lemon grass leaf extract when compared with theuntreated control. Neem extract was the most effective while lemon grass extract was the least. Crude extracts from all the plant leaves tested increased seed germination and seedling emergence of African yam bean seeds and gave significant (P. 0.05) reduction of mycelial growth of all the fungi tested when compared with their aqueous extracts. Leaf extracts of neem, basil, bitter leaf and paw-paw, which are cheap and environmentally safe, are promising for protecting African yam bean seeds against major seed-borne fungi and in the improvement of the crop.

Drug resistance in malaria infection is a serious public health challenge. Thus, scientific search for alternative treatment measures among the local medicinal plants is exigent. We therefore investigated the anti-plasmodial efficacy and genotoxicity of the methanolic leaf and stem extracts of Alstonia plant at varying concentration (200 mg/kg, 400 mg/kg and 600 mg/kg) in mice infected with chloroquine sensitive Plasmodium berghei. The phytochemical screening of the extract revealed that leaf sample contained significantly higher secondary metabolites, except saponins (p<0.05). Anti-plasmodial activities of the two extracts were duration and dose- dependent. Stem bark extract showed higher curative potential with inhibition rate of 56.71% at 400 mg/kg whereas, leaf extract was efficient at 600mg/kg with 52.15% inhibition rate. Stem bark extract at 400 mg/kg improved the enzymatic activities of the mice; it lowered serum ALT (6.88±4.42) and increased liver ALT (41.07±5.56). Similarly, 400 mg/kg leaf extract showed highest AST (70.65±4.00) and ALT (44.65±7.83) activities in the kidney and liver respectively. Analysis of genotoxicity revealed that micronucleus and abnormal (binucleated, notched and blebbed) were prevalent among the experimental mice which increased significantly (p<0.05) at all concentrations except at 600mg/kg leaf extract. Therefore, this present study indicates that both leaf and stem bark extracts of A. boonei possess anti-plasmodial activity and are less genotoxic when compared with standard drug.Keywords: Genotoxicity; Plasmodium berghei; metabolites; micronucleus; blebbed; standard drug

Introduction: One of the vectors of disease spread is flies. House flies (Musca domestica) are often found alive in almost all types of environments. Several previous studies have shown that pandanus leaves (Pandanus amaryllifolius), basil leaves (Ocimum sanctum), and bay leaves (Syzygium polyanthum) contain active compounds that are toxic to insects, but no one has compared the effectiveness of the four types of plants. This research aims to determine the effectiveness of administering extracts from various plant leaves as a bioinsecticide for houseflies (Musca domestica). Methods: This type of research is experimental using a completely randomized design with 6 treatments. The treatment consisted of administering Pandan Leaf extract (Pandanus amaryllifolius), Basil Leaf extract (Ocimum sanctum) and Laurel Leaf extract (Syzygium polyanthum) each with a concentration of 5 ml, 10 ml and 15 ml and using a control for comparison. Observations were carried out for 24 hours observing the number of dead flies. Results: The results of this study indicate that pandan leaf extract (Pandanus amaryllifolius), basil leaf extract (Ocimum sanctum), and bay leaf extract (Syzygium polyanthum) are effective in killing houseflies (Musca domestica). The most effective concentration being 15 ml. Research Implications: This research demonstrates that the use of natural bioinsecticides from plant extracts can control the population of house flies (Musca domestica) so that the risk of disease transmission in the community can be significantly minimized Conclusion: Pandan leaf extract (Pandanus amaryllifolius), basil leaf extract (Ocimum sanctum) and bay leaf extract (Syzygium polyanthum) can be used as natural alternative ingredients to exterminate houseflies (Musca domestica) Limitations of the research: This research did not control environmental conditions such as sunlight, temperature, and humidity. Other than that, neither the effects on fly resistance nor the effects on non-target organisms have been studied. Recommendations for Future Research: It is hoped that future research can examine the effects of fly resistance and killing on non-target organisms and control environmental conditions.

The purpose of the study was to determine the effect of the appropriate and effective application of the L. lecanii fungus and C. odorata L. leaf extract in increasing the mortality of A. gossypii. The study was divided into two stages. The first stage was in vitro test of L. lecanii synergism plus C. odorata L. leaf extract in PDA media with four treatments, namely LK0= L. lecanii 107 conidia.mL-1 plus 0% C. odorata L. leaf extract, LK1= L. lecanii 107 conidia.mL-1 plus 10% C. odorata L. leaf extract, LK2= L. lecanii 107 conidia.mL-1 plus 25% C. odorata L. leaf extract; LK3= L. lecanii 107 conidia.mL-1 plus 40% C. odorata L. leaf extract. The second stage was the toxicity test of the application of L. lecanii suspension and C. odorata L. leaf extract on mortality of A. gossypii. The toxicity test was based on the results of the synergism test, where the addition of C. odorata L. leaf extract to L. lecanii growing media showed incompatible results. Therefore, the toxicity test was carried out separately with five treatments, namely H0= Control (aqua dest); H1= Conidia suspension L. lecanii 107 conidia mL-1; H2= 10% C. odorata leaf extract; H3= 25% leaf extract of C. odorata; H4= 40% C. odorata leaf extract. The results showed that the compatibility test of L. lecanii with leaf extract of C. odorata L. was incompatible and classified as toxic. The addition of C. odorata L. leaf extract in concentrations of 10%, 25%, and 40% could significantly inhibit colony growth, sporulation, and conidia viability of L. lecanii, with a higher level of inhibition as the concentration of C. odorata L. leaf extract, was added. The toxicity test of a separate application of C. odorata L. leaf extract and L. lecanii suspension had a significant effect on mortality of 3rd instar nymph A. gossypii, with the highest mean mortality found in a single application of 40% C. odorata L. leaf extract with an average mortality of 100% at 96 HAA(Hours After Application) observations. Keywords: A. gossypii, C. odorata L. leaf extract, L. lecanii, synergism.

Highlight : Patients experiencing failure of first-line drug and developing multidrug resistant tuberculosis (MDR-TB) has increased throughout 2011-2015. Castor plant leaf extract (Ricinus communis L.) has an antibacterial potential against the growth of Mycobacterium tuberculosis bacteria. Abstract: Mycobacterium tuberculosis is the cause of pulmonary tuberculosis that can reduce human health. In the therapy of the disease, patients can develop resistance to tuberculosis drugs. Based on the 2015 health profiles of Indonesia, 15,380 people were suspected to have multidrug-resistant tuberculosis (MDR-TB), while 1,860 people were confirmed patients with MDR-TB. There is a need for innovation to develop the latest treatments using natural ingredients, one of which is castor plant (Ricinus communis L.) that contains antibacterial compounds against Mycobacterium tuberculosis. This study aimed to understand the antimicrobial potential of castor plant (Ricinus communis L.) leaf extract against the growth of Mycobacterium tuberculosis. This scientific paper was a quantitative systematic review study. Literature in the form of journal articles and books were obtained through search engines, i.e. ebook database, Google Scholar, Cochrane, Wiley, and PubMed. The results of the literature source search were 19 journal articles and 4 ebooks, as well as 4 journal articles that were in accordance with the title of this literature review and discussed the effects of castor plants on the growth of Mycobacterium tuberculosis. The results of the analysis showed that castor plant (Ricinus communis L.) leaf extract has the potential in the antibacterial activity against the growth of Mycobacterium tuberculosis because it contains phytochemicals in the form of flavonoids, saponins, alkaloids, tannins, and fatty acid amides derived from ricinoleic acid as the main constituent of castor plants (Ricinus communis L.). There is antimicrobial potential for castor plant (Ricinus communis L.) leaf extract against the growth of Mycobacterium tuberculosis.

Sulphur treatment alters the therapeutic potency of alliin obtained from garlic leaf extract

The present study was designated to evaluate the antifungal activity and to root out the antifungal plant leaf extracts from this Indian folk-flore. The in vitro antifungal assay was performed by agar diffusion test and minimum inhibitory concentration (MIC) for hexane, ethyl acetate, methanol and distilled water plant leaf extracts. Extraction of 17 different plant leaves was carried out in different solvents such as hexane, ethyl acetate, methanol and distilled water. Among them extractive yield of methanol was maximum than the rest of the three solvents. These extracts were screened for their antifungal activity against nine different fungi. Among these ethyl acetate extracts of Adhatoda vasica, Ocimum sanctum and Holoptelea integrifolia exhibited maximum antifungal activity against Alternaria sp., Aspergillus parasi, Aspergillus nidulans, Trichoderma harzianum and Aspergillus flavus with MIC of 80, 40 and 20 ppm against Aspergillus nidulans and Alternaria sp. Ethyl acetate extracts showed promising antifungal activity against Adhatoda vasica, Ocimum sanctum and Holoptelea integrifolia against Aspergillus nidulans, and Alternaria sp. might be applicable as fungicide against fungal plants disease.

Antimalarial resistance is a major challenge for effective control of malaria. This triggered the need to monitor the efficacy of Azadirachta indica and Ocimum gratissimum in the disease management. The present study compared antimalarial activities of the leaf extracts of the individual plants and evaluated their combined effects on malaria infected mice. The leaves of the different plants were extracted with absolute ethanol (BDL 95%) for the test. Clean albino mice were experimentally infected intraperitoneally with chloroquine-sensitive Plasmodium berghei Nk65 strain. Parasitaemia level was determined before parasite inoculation and at 24 hours post treatment period. Efficacy of the leaf extracts was tested on the infected mice using Peter’s 4-days suppressive and curative tests, and secondary biological assessment procedures.The lethal median dose (LD50) recorded for neem and clove basil leaf extracts were 31.62 and 1246.9 mg/kg body weight, respectively. Infected mice treated with leaf extracts of the plants and their combinations produced significant dose- dependent activity against the parasite (P&lt;0.05). Highest reduction of parasitaemia was observed on day 4. Maximum parasitaemia reduction (78.65%) was attained with 30mg/kg of the combination of the extracts on the 7th day. These observations indicate better anti-malarial activity of the combination therapy as compared with the individual extracts of the plants, and also show their good antimalarial potentials.

In vitro antibacterial activity of methanol and ethanol leaf extracts of Euphorbia heterophylla and Pterocarpus lucens were investigated against six bacterial clinical isolates using the tube dilution and agar diffusion methods. Salmonella typhi was the most susceptible to methanol leaf extracts of E. heterophylla with a zone of inhibition ranging from 16 to 24 mm for 12.5 to 100 mg/ml concentration. This was followed by Streptococcus lactis, Escherichia coli, Staphylococcus aureus, and Shigella species in that order with Proteus vulgaris not susceptible to the different test concentrations of both plant extracts. E. heterophylla had the least minimum inhibitory concentration (MIC) of 6.25 mg/ml against E. coli and S. typhi while P. lucens extract MIC of 25.00 mg/ml was the least against S. typhi. Since there is an inverse relationship between MIC value and susceptibility of the clinical test isolates, the MIC values also shows that E. heterophylla methanol leaf extracts were more potent to the susceptible test organisms having lower MIC values than the corresponding ethanol leaf extract MIC value. E. heterophylla extract minimum bactericidal concentration (MBC) was 25.00 mg/ml for the sensitive isolates except for methanol extract with 12.50 mg/ml against S. typhi and ethanol extract with 12.50 mg/ml against S. aureus. P. lucens extract MBC was 100.00 mg/ml for the sensitive test isolates except for ethanol leaf extracts with 50.00 mg/ml against S. typhi. The killing rate of E. heterophylla methanol leaf extract MBC shows that E. coli was most rapidly killed at a rate of 4.53 &times; 106 CFU/min with S. aureus as the least killed at a rate of 0.62 &times; 106 CFU/min. S. lactis and E. coli were the most rapidly killed by P. lucens leaf extract MBC at a rate of 1.90 &times; 106 CFU/min. The killing rate of the extracts showed a positive support in the potential use of these plants in curing some infections as done by the traditional herbal healers in Anyigba, Kogi State, Nigeria.&nbsp;&nbsp;&nbsp;&nbsp; &nbsp; Key words: In vitro, antibacteria, extract, inhibitory.In vitro antibacterial activity of methanol and ethanol leaf extracts of Euphorbia heterophylla and Pterocarpus lucens were investigated against six bacterial clinical isolates using the tube dilution and agar diffusion methods. Salmonella typhi was the most susceptible to methanol leaf extracts of E. heterophylla with a zone of inhibition ranging from 16 to 24 mm for 12.5 to 100 mg/ml concentration. This was followed by Streptococcus lactis, Escherichia coli, Staphylococcus aureus, and Shigella species in that order with Proteus vulgaris not susceptible to the different test concentrations of both plant extracts. E. heterophylla had the least minimum inhibitory concentration (MIC) of 6.25 mg/ml against E. coli and S. typhi while P. lucens extract MIC of 25.00 mg/ml was the least against S. typhi. Since there is an inverse relationship between MIC value and susceptibility of the clinical test isolates, the MIC values also shows that E. heterophylla methanol leaf extracts were more potent to the susceptible test organisms having lower MIC values than the corresponding ethanol leaf extract MIC value. E. heterophylla extract minimum bactericidal concentration (MBC) was 25.00 mg/ml for the sensitive isolates except for methanol extract with 12.50 mg/ml against S. typhi and ethanol extract with 12.50 mg/ml against S. aureus. P. lucens extract MBC was 100.00 mg/ml for the sensitive test isolates except for ethanol leaf extracts with 50.00 mg/ml against S. typhi. The killing rate of E. heterophylla methanol leaf extract MBC shows that E. coli was most rapidly killed at a rate of 4.53 &times; 106 CFU/min with S. aureus as the least killed at a rate of 0.62 &times; 106 CFU/min. S. lactis and E. coli were the most rapidly killed by P. lucens leaf extract MBC at a rate of 1.90 &times; 106 CFU/min. The killing rate of the extracts showed a positive support in the potential use of these plants in curing some infections as done by the traditional herbal healers in Anyigba, Kogi State, Nigeria. &nbsp; &nbsp; &nbsp; Key words: In vitro, antibacteria, extract, inhibitory.

Fusarium oxysporum f. sp. zingiberi is a soil-borne plant pathogen causing rhizome rot on ginger. This pathogen can survive in the soil for several years without a host plant. This study aimed to examine the viability and virulence of 21 isolates of F. oxysporum f. sp. zingiberi after being preserved for 17 years in sterile soil. Fungal viability was determined by descriptive method, while the experiment using randomized block design was conducted to examine the virulence of fungal isolates. The treatments consisted of control, 21 isolates of F. oxysporum f. sp. zingiberi from Boyolali and Temanggung, each treatment was replicated three times. The variables consisted of colony colors and diameters, macroconidia and microconidia shapes, growth time, dry weight of mycelia, conidia density, incubation period, affected area, rhizome wet weight difference, and waste index. The results showed that all fungal isolates which were stored in sterile soil for 17 years still had the ability to grow well on PDA medium and fill up petri dishes in 11–36 days. Moreover, all the isolates caused infection and disease symptoms development in ginger rhizome var. Gajah. Less virulence isolate was characterized by a long incubation period (6–12 days after inoculation) and smaller affected area of the rhizome.