Leucaena Leucocephala Research Articles

Leucaena-Based Alley Cropping System: An Approach for Reclaiming Degraded Land, Reducing the Use of Inorganic Nitrogen Fertilizer, and Improving Crop Productivity

Alley cropping, an agroforestry system that integrates trees and arable crops, holds the potential to improve both crop yields and soil health. It has been found to be effective for upland crops in many countries of the world. However, the utilization of alley cropping to improve soil health in the terrace ecosystem of Bangladesh is poorly understood. Therefore, this study was undertaken to assess the changes in soil biochemical properties and quantify the cabbage yield under three alley widths of Leucaena leucocephala (3.0, 4.5, and 6.0 m size) and five nitrogen (N) levels [0, 40, 80, 120, and 160 kg N ha‒1 (0, 25, 50, 75, and 100% of recommended N rates, respectively) with the addition of pruned materials of L. leucocephala (Ipil-ipil)]. The field experiment was conducted following a split-plot design, where alley width was considered as the main-plot factor and N rate as the sub-plot factor. Within each main plot, the five N rates were replicated thrice. Control plots with similar N doses were applied accordingly without addition of pruned materials to compare the results with alley cropping. Data were collected on the biochemical properties of the soil [soil pH, organic carbon (C), total N, available phosphorus (P), exchangeable potassium (K), microbial biomass C, and biomass N] and the yield of cabbage quantified [edible head weight (kg plant‒1) and head yield (t ha‒1)] under different alley widths and control. Findings revealed that organic C, total N, available P, exchangeable K, microbial biomass C, and biomass N in the topsoil exhibited maximum values in the L. leucocephala-based alley plot, which is proved to be a possible solution of restoration of degradable land. Additionally, L. leucocephala-based alley cropping improved the soil pH, indicating a potential avenue for more-sustainable land management practices. Results also showed that alley widths and N rates have significant effects on cabbage (Brassica oleracea L. var. capitata) yield. Alley width of 6.0 m along with 100% N provided the highest cabbage yield followed by 75% N in 6.0 m alley, and the control with 100%. The wider alley minimizes tree–crop competition, allowing for optimal cabbage production. These aforementioned results suggest that alley cropping with L. leucocephala is a promising approach to enhance soil fertility and crop productivity in the terrace ecosystem of Bangladesh.

Agroforestry adoption and its influence on soil quality under smallholder maize production systems in western Kenya.

Agroforestry, a sustainable land use practice, was introduced in western Kenya in the early 1990sas a soil fertility replenishment strategy. Since then, the effect of the practice on soil quality has seldom been evidenced. A study was conducted to assess the impact of agroforestry adoption on soil quality under smallholder maize systems in the region. Soil samples were collected from two land use practices: agroforestry adoption (90) and non-agroforestry adoption (30) at 0-30 cm depth from two locations (Busia and Kakamega counties). Results showed variations in soil characteristics between the study locations. Soils in Kakamega contained higher concentrations of soil organic carbon (SOC), exchangeable cations, and micronutrients. On average, agroforestry adoption significantly (P ≤ 0.05) improved soil physicochemical properties. Under agroforestry adoption, bulk density (BD) reduced by 21% (from 1.4 to1.1 g cm-3), while (SOC) increased by 75% (0.8-1.4%), P by 80% (3.0-5.4 mg kg-1), exchangeable K by 256% (0.3-8.0 cmol kg-1), Ca by 100% (1.0-2.0 cmol kg-1), S by 50% (0.2-0.3 mg kg-1), and Cu by 18% (2.8-3.3 mg kg-1). Agroforestry adoption significantly increased K and Cu levels above the critical thresholds of 0.4 cmol kg-1 and 1.0 mg kg-1, respectively, at both locations. Sesbania sesban and Leucaena leucocephala influenced soil BD, pH and P (4.3.-7.0 mg kg -1), exchangeable K (0.4-0.7 cmol kg-1), Mg (0.1-0.2 cmol kg-1), and Mn (13.5-25.2 mg kg-1) at both locations, while Calliandra calothyrsus significantly increased SOC in Kakamega only. These findings highlight the significance of agroforestry in soil fertility management. Furthermore, Sesbania proved to be effective in enhancing the characteristics of soils at both sites, hence, the need for its inclusion in agroforestry extension messages. Further studies are needed to understand nutrient release mechanisms from agroforestry biomass and their influence on soil characteristics and maize yield in smallholder farming systems.

Anthelmintic activity of Leucaena leucocephala var. tarramba leaf extracts against ruminant gastrointestinal parasites

Abstract With the growing resistance of parasites to most currently available anthelmintics and increasing concerns about anthelmintic residues in animal products and the environment, there is a heightened interest in exploring plant-based alternatives. This study aims to evaluate the anthelmintic properties of Leucaena leucocephala var. tarramba leaf extracts against gastrointestinal parasites in ruminants, examining their effects on internal parasites in vitro and in vivo. For the in vitro analysis, eggs isolated from fresh fecal samples of worm-infected goats were incubated with varying concentrations (0.25% to 1%) of the leaf extract. The best extract concentration in the in vitro test was then selected and formulated into a bolus for use in the in vivo test on ten cattle aged 7 to 12 months. The cattle were randomized into two groups: treatment and control, with the treatment group receiving the leaf extract and the control group receiving a commercial anthelmintic. In vitro tests revealed that a 1% concentration of Tarramba leaf extract significantly inhibited egg hatching, showing comparable efficacy to commercial anthelmintics. In vivo tests on Bali cattle demonstrated that the extract reduced parasite prevalence by 60% over eight weeks, though this was less effective than the 80% reduction achieved with commercial anthelmintics. These findings suggest that Leucaena leucocephala var. tarramba could offer a sustainable alternative to synthetic anthelmintics, reducing the environmental impact and risk of drug resistance in parasite control for livestock.

Correction: The soil response in experimental Leucaena leucocephala plots under manipulated rainfall regimes

Correction: The soil response in experimental Leucaena leucocephala plots under manipulated rainfall regimes

Molecular cloning & In silico Analysis of Metallothionein Type II Gene from Leucaena leucocephala (a Leguminous Species)

Plant metallothioneins (MTs) are low molecular weight cysteine rich proteins consisting of a single polypeptide chain with two thiol-rich sites - α and β domain which have a strong affinity to metal ions. Apart from heavy metal detoxification, plants MTs are involved in regulation of essential metals and have been shown to be responsive toward other abiotic stresses. Some plants have evolved physiological and molecular mechanisms like compartmentalisation, sequestration, chelation or exclusion against metal stress. Leucaena leucocephala, a leguminous tree species has heavy metal tolerance and survival ability in inhospitable conditions. In this study, Metallothionein type II (LlMTII) gene was isolated from L. leucocephala and cloned. Sequencing revealed the Open Reading Frame of 228 bp in length which encoded putative protein composed of 75 amino acids with a molecular mass of 7.671 kDa and theoretical pI of 4.90. Negative GRAVY number indicated hydrophilic nature of the protein, aliphatic and instability indices showed unstable nature in in-vitro conditions. LlMTIIbelonged to p1 sub-family of Metallothioneins and 3D modeling showed two Cys-rich domains with a long Cys-free linker of about 44 aa. It exhibited association with various abiotic stress responsive transcription factors. Multiple sequence alignment and phylogenetic analysis showed high level of similarity in the conserved domains especially Cys residues with many species (both Monocots and Dicots). This study helps us to better understand LlMTII gene and its role in heavy metal detoxification and conferring tolerance to other abiotic stresses.

Unveiling Fusarium falciforme: Genome sequencing of a Novel wilt causing pathogen in subabul (Leucaena leucocephala L.) in India.

Unveiling Fusarium falciforme: Genome sequencing of a Novel wilt causing pathogen in subabul (Leucaena leucocephala L.) in India.

Effects of Hardwood Biomass Variability on Biochar Properties: Insights from Wood Waste Utilization

Wood is an abundant and renewable resource with considerable potential for upcycling into high-value products such as biochar, thereby facilitating the conversion of waste into useful materials. The structural and compositional diversity among wood species significantly influences the properties of the resulting biochar. This study aimed to investigate the properties of biochar derived from five distinct hardwood species, all sourced from the same tropical region and produced under standardized pyrolysis conditions using a patented furnace. The hardwood species examined were Leucaena (Leucaena leucocephala (Lamk.) de Wit.), paper flower climber (Getonia floribunda (Roxb.) Lam.), rain tree (Samanea saman (Jacq.) Merr.), climbing wattle (Albizia myriophylla Benth.), and Siamese blachia (Blachia siamensisGagnep.). The investigation focused on fundamental properties of the biochar, including morphology, elemental composition (C, H, N, O), water-holding capacity, aromaticity (H/C ratio), polarity (O/C and (O+N)/C ratios), and the C/N ratio. Results indicated that all five biochar types were of high quality, with carbon contents ranging from 66.64% to 84.76%, a high degree of aromaticity (H/C < 0.7), low polarity, and enhanced stability. The biochars exhibited a range of pore structures from macropores to mesopores, with pore volumes of 0.010–0.074 cm³/g and specific surface areas ranging from 2.92 m²/g to 144.59 m²/g. These structural attributes influence their water-holding capacities; however, the relationships among polarity, pore volume, pore size, and water-holding capacity remain unresolved. The findings highlighted substantial variability in the properties and morphology of the biochars, despite their production under identical conditions. This variability underscores the need for further in-depth studies to elucidate the factors governing biochar properties and to enable the tailoring of biochar for specific applications. Doi: 10.28991/HEF-2025-06-01-02 Full Text: PDF

Insect Diversity of Mangrove Ecosystems in Beras Basah Village, Langkat, North Sumatra, Indonesia

This study aimed to investigate the diversity of insects, including stingless bees (Heterotrigona) in the mangrove ecosystem of Beras Basah Village, Langkat Regency, North Sumatra, Indonesia. The mangrove plantation in Beras Basah Village contained 11 species, including Rhizophora apiculata, Rhizophora mucronata, Rhizophora. stylosa, Avicennia marina, Sonneratia alba, Bruguiera gymnorhizza, Xylocarpus granatum, Excoecaria agallocha, Scyphiphora hydrophyllacea, Lumnitzera racemosa, and Nypa fruticans. Additionally, five flowering plant species were identified in the vicinity: Antigonon leptopus, Asclepias curassavica, Helianthus annuus, Calliandra grandiflora, Portulaca grandiflora. Other plant species recorded in the area included Cocos nucifera, Pandanus amaryllifolius, Leucaena leucocephala, Ziziphus mauritiana, Albizia chinensis, and Terminalia catappa. Insect identification from four observations using sweep nets, yellow sticky traps, and blue pan traps resulted in the collection of 1,525 individual insects belonging to eight orders, 17 families, and 19 genera. These insects included ten predators and pests, comprising five Odonata species (Neurothemis terminata, Orthetrum sabina, Orthetrum testaceum, Potamarcha congener, Zyxomma obtusum) and five ant species (Camponotus spp., Crematogaster spp., Nylanderia spp., Odontoponera denticulate, Pheidole spp.). The insect functional status within the mangrove area was determined to include seven species of scavengers, 12 species of herbivores, two species of pollinators, 19 species of predators, one species of parasitoid, and four species of insect disease vectors. This study provides valuable insights into the insect diversity and ecological roles within the mangrove ecosystem of Beras Basah Village, contributing to a better understanding of the conservation and management of this important habitat.

Tracing the Wanderers of the Riverbed: Home Ranges, Daily Movements and Habitat Frequentation of the Grey Francolin in Reunion Island

ABSTRACTThe Grey Francolin, a Phasianidae species native to India and Pakistan, was introduced to Reunion Island in the XVIIIth century for hunting. One of its populations resides in the Rivière des Pluies riverbed, in the island's north, a secondary vegetation area surrounded by an industrial zone. The study aimed to determine the minimum daily movements, home range size, and macro‐habitat use of individuals during the breeding season, as well as the species used for roosting at night. Six individuals from four different pairs were fitted with GPS. Home ranges ranged from 2.4 to 5.8 ha (MCP 100) and from 2.1 to 8.3 ha (Kernel 95), with averages of 4.5 ± 1.2 (MCP 100) and 5.0 ± 3.0 (Kernel 95). Minimum daily movements varied from 252 to 513 m, averaging 402 ± 147 m per day. GPS points were found in three macro‐habitats: wooded areas with Pithecellobium dulce and Leucaena leucocephala (62%), pebble riverbed (23%), herbaceous vegetation (13%), but never in the urban zone.

Effects of processed <i>Leucaena leucocephala</i> Leaf Meal on the Reproductive and Pre-weaning Performance of Rabbits (<i>Oryctolagus cunniculus</i>)

In an experiment designed to determine the effects of dietary inclusion of differently processed Leucaena leucocephala leaf meal on reproductive performance of rabbits does, 40 local breeds does of about 20 weeks old were fed five diets (T1-T5). Four different processing methods were used to reduce the mimosine content in Leucaena leucocephala leaf meal for inclusion in diets T2, T3, T4 and T5 at 40% fixed level. Diet T1 which is the control contained no Leucaena leucocephala leaf meal and was designated NLLM, T2 contained air dried Leucaena leucocephala leaf meal and was designated ADLLM, T3 contained soaked Leucaena leucocephala leaf meal in fresh water for 36 hours and was designated FWLLM, T4 contained Leucaena leucocephala leaf meal soaked in 600C hot water for 24 hours and was designated HWLLM while T5 contained fermented Leucaena leucocephala leaf meal for five days and was designated FLLM. The result showed significant (P<0.05) differences in all the reproductive parameters measured. Pre weaning performance traits of rabbit kittens were significantly (P<0.05) lower in rabbits fed diets containing air dried Leucaena leucocephala leaf meal. It was concluded that air dry method of processing Leucaena leucocephala leaf meal was not effective enough to reduce the mimosine content to a tolerable level in rabbit diet. Soaking Leucaena leucocephala leaf meal, in fresh water for 36 hours at room temperature, in 600C hot water for 24 hours and fermentation of Leucaena leucocephala leaf meal for five days were recommended for use at 40% level of inclusion in rabbit diet.

Assessment of metal residues in soil and evaluate the plant accumulation in copper mine tailings of Dongchuan, Southwest China.

This study aimed to identify suitable plants for remediating metal pollution in copper tailing soil and supporting ecological reclamation in Dongchuan, Yunnan, focusing on three major mining regions: Tangdan, Yinmin, and Lanniping. The Nemerow comprehensive pollution index was employed to evaluate the metal contamination levels, and the enrichment and transfer capacities of the dominant plants for Cd, Cu, Pb, and Zn were analyzed to identify remediation candidates. The findings revealed severe pollution in the copper tailing soil, with Cu as the primary pollutant, and with a pollution rate of 77.778%. A total of 96 plant species from 42 families and 87 genera were recorded, including 29 dominant species across 17 families and 26 genera, with herbs comprising 62.068% of the dominant types. Among the tested plants, Leucaena leucocephala, Rumex acetosa, Festuca rubra, and Salix balfouriana exhibited significantly higher metal enrichment and transfer capacities, rendering them for ecological restoration. Salix balfouriana with the highest comprehensive membership function score of 5.298 was identified as the preferred species for ecological restoration in the Dongchuan Cu tailing area. Correlation analysis revealed a positive relationship between the metal content in underground plant parts and both the total metal content and organic matter (OM) in the rhizosphere soil, whereas a negative correlation was observed with soil pH. The soil in the Dongchuan Cu tailing area is severely contaminated by metals, mainly Cu. Among the identified plants, S. balfouriana emerged as the most suitable candidate for metal accumulation. This study establishes a comprehensive theoretical framework for the application and promotion of phytoremediation technology in the Dongchuan copper tailings area.

The soil response in experimental Leucaena leucocephala plots under manipulated rainfall regimes

The soil response in experimental Leucaena leucocephala plots under manipulated rainfall regimes

Correction to: Dryer Pantanal wetlands may increase Leucaena leucocephala (Fabaceae) invasion

Correction to: Dryer Pantanal wetlands may increase Leucaena leucocephala (Fabaceae) invasion

Effect of the addition of fermented Lamtoro leaf water extract (Leucaena Leucocephala) in drinking water on the performance and histology of the broiler intestine

Effect of the addition of fermented Lamtoro leaf water extract (Leucaena Leucocephala) in drinking water on the performance and histology of the broiler intestine

Bioactive Compounds and Antioxidant Activity of Leucaena leucocephala Leaves Collected in Côte d’Ivoire

Aims: In a context of increasing valorization of tropical plant resources as potential sources of bioactive compounds, Leucaena leucocephala, a pantropical legume widely distributed in Côte d'Ivoire, represents a model of interest still insufficiently explored for its phytochemical and antioxidant properties. The present study aimed to determine the contents of condensed tannins, total tannins and total polyphenols, as well as the antioxidant activity of Leucaena leucocephala leaf extracts, in order to valorizing its nutraceutical potential. Study Design: Leucaena leucocephala leaves were collected at the vegetative stage in the locality of Yamoussoukro, dried under shelter at room temperature and then crushed. The powder obtained was used for the various analyses at the Chemistry laboratory of the Institut National Polytechnique Félix HOUPHOUËT-B0IGNY in Yamoussoukro. Place and Duration of Studies: After the harvest, the analyses were carried out from March to June 2023. Methodology: Maceration and decoction were used to extract the phenolic compounds. Maceration extraction was done using a mixture of distilled water/ethanol (30/70, v/v) and distilled water/methanol (30/70, v/v) as solvent while decoction was done using distilled water only. Results: The condensed tannin contents obtained in Leucaena leucocephala leaves were 137.52 ± 92.74, 140.22 ± 92.74, and 32.63 ± 7.13 mg Cat E/g extract, for methanolic, ethanolic and aqueous solutions, respectively. The total polyphenols contents were 5.74 ± 0.22, 4.01 ± 0.22, and 1.32 ± 0.11 mg GAE/g extract, respectively. The total tannin contents were 209.05 ± 7.49, 207.92 ± 7.49, and 55.85 ± 7.49 mg TAE/g extract, respectively. The antioxidant activities were 30.99±1.95, 19.75±0.740, and 12.21±2.63 mg TE/g extract, respectively. Conclusion: It was concluded that Leucaena leucocephala leaves collected in Côte d’Ivoire contain bioactive compounds and has antioxidant activity with variations depending on the extraction solvent.

Uses and protein content of green guaje seeds (Leucaena leucocephala (Lam.) de Wit offered in the markets of Southern Puebla State, Mexico

Leucaena leucocephala, commonly known as guaje, is a woody plant endemic to Mexico with edible seed pods. Despite its historical significance as a resource for Mesoamerican peoples, the nutritional value of guaje remains poorly documented. This study aims to explore the uses and protein content of green guaje seeds offered in markets in Southern Puebla State, Mexico. The research involved interviewing vendors at various markets in Southern Puebla, including Tepeaca tianguis, Revolución market, La Rotonda market, and El Moralillo tianguis, where a total of 127 stalls were surveyed. Information was gathered regarding the source, uses, and preparation of guaje seeds. Additionally, the protein content of the seeds was analyzed to determine its nutritional value. Green guaje seeds were available at diverse sales points, with distinct vendor profiles: regatoneros (local resellers) predominantly at the Revolución and La Rotonda markets, while self-producing vendors were more common at Tepeaca tianguis. Eleven culinary preparations featuring guaje were identified, with guaxmole being the most prevalent (40.66%) and various chili sauces with guaje following (38.52%). Although most vendors were unaware of the medicinal uses of guaje seeds, a few recognized their traditional applications, including as a dewormer and for alleviating coughs, stomach pains, and cramps. The protein content in guaje seeds varied significantly, ranging from 6.991 µg/µl in seeds from San Pablo Zoyatitlanapan at the Revolución market to 22.3766 µg/µl in seeds from Acatzingo at the Tepeaca tianguis. This study highlights the underutilized potential of guaje seeds and provides valuable information on their protein content and culinary uses. The significant variability in protein content based on the seed's origin underscores the need for further research into factors affecting nutritional quality. Additionally, increased awareness of guaje's medicinal properties could enhance its utilization and benefits in local communities.

Chemical Composition and Relative Feed Value of Tree Legume Leaves as a Feed Source for Ruminants

Background: Tree legumes are a pivotal to meet the nutritional needs to support the productivity of ruminants. The aim of this study was to evaluate the chemical composition and relative feed value of various types of tropical tree legumes as a potential feed source for ruminants. Methods: The study was designed using a Completely Randomized Design (CRD) with four treatments for legume tree species and three replications. A1 (Sesbania grandiflora), A2 (Leucaena leucocephala), A3 (Calliandra calothyrsus) and A4 (Moringa oleifera). Result: Variance analysis showed that the species of tree legume had a significant effect (P less than 0.05) on the content of crude protein (CP), crude fibre (CF), nitrogen-free extract (NFE), digestibility of dry matters (DMD), dry matter intake (DMI), relative feed value (RFV) and not significantly different (P greater than 0.05) on EE (EE). The four tree legumes had protein content above 20.20%, CF below 21.58%, EE below 7.88% and NFE above 33.48%, DMD above 65.00%, DMI above 3.4% of body weight, RFV above 182.74 including good quality feed ingredients. The four tree legumes are potential ruminant feed in the tropics.

In silico approach to predict antidiabetic activity of bioactive peptide from Leucaena leucocephala seed during gastrointestinal digestion

Abstract Leucaena leucocephala (leucaena) was an easy growing species with year-round flowering and fruiting and abundant seed production. Its mature seed contained 30.81%-50.37% protein that potential to be explored as bioactive peptide source, as well as nutrition source. Previous study showed that the seed extract has antidiabetic activity. However, study of bioactive peptide role on antidiabetic activity of leucaena was limited. In silico approach recently was widely used to predict the potential of bioactive peptide derived from certain protein source due to its supporting result to laboratory experimental approach. This study aimed to investigate the antidiabetic potential of bioactive peptide of leucaena’s protein during gastrointestinal digestion. In silico digestion and toxicity prediction were performed using BIOPEP and ToxinPred online tools, respectively. After in silico digestion using pepsin, trypsin and chymotrypsin, the result showed that leucaena seed was a potential source of bioactive peptide with DPP-4 inhibitor, alpha glucosidase inhibitor, glucose absorption stimulation and antioxidative activities. The potential peptide with biological activity was predicted as safe.

Performance of Balinese Pigs Fed with Banana Stem and Lamtoro (Leucaena leucocephala) Leaf-Based Diets Fermented for Different Durations in the Sukanadi Women Farmer Group

Balinese pigs are one of the livestock commodities that the community has raised. However, their maintenance is still carried out traditionally by utilizing makeshift feed ingredients in the vicinity, so the appearance of Balinese pigs is not optimal with indicators, and their growth is still below the NRC standard. With simple maintenance, the weight gain of local pigs can only be achieved 180-260 gr/head/day, which is still below the NRC standard of 500-750 gr/head/day. Therefore, efforts must be made to increase weight gain by improving feed quality through fermentation. This study aims to improve the performance of local Balinese pigs by treating feed with various fermentation times from surrounding materials such as banana stems, forage, and complete feed mixtures (rice bran, fine corn, and concentrates). This study used a complete randomized design with fermented feed treatment with variations in fermentation time, namely feed without fermentation (F0), feed fermented for 14 days (F1), and feed fermented for 28 days (F2). Each treatment used 3 Balinese/local pigs of 2-3 months old weaning age with a weight range of 10-12 kg. The banana stem and lamtoro leaf-based feedstuffs fermented for 28 days showed significant results or the best on the performance of Balinese pigs. Contribution to Sustainable Development Goals (SDGs):SDG 1: No PovertySDG 2: Zero hunger SDG 9: Industry, Innovation, and Infrastructure SDG 12: Responsible Consumption and Production SDG 13: Climate ActionSDG 15: Life on Land

Palatability level, analysis of types and phytochemical content of feed on long-tailed monkeys (Macaca fascicularis Raffles, 1821) in the Kaliurang nature tourism forest, Yogyakarta

The forest area on the slopes of Mount Merapi, located in the northern part of Yogyakarta, has a lot of flora and fauna. One of the fauna that lives in the area is a long-tailed monkey (Macaca fascicularis Raffles 1821). Long-tailed monkeys eat lots of fruit and are grouped into frugivorous animals. However, long-tailed monkeys, such as leaves and flowers, will have more abundant food in their habitat. Therefore, this study analyzed long-tailed monkey feed to see the percentage of long-tailed monkeys who chose feed. In addition, phytochemical content analysis was conducted as supplementary data to determine the feeding preferences of long-tailed monkeys. The data obtained were analyzed descriptively. The results showed that the plant species selected by the long-tailed monkeys in the Tlogo Muncar and Tlogo Nirmolo areas were Cinchona pubescens Vahl, Leucaena leucocephala (Lam.) de Wit, Melastoma polyanthum Burm. f, Carex flaca and Sp 1. The plant parts that long-tailed monkeys ate were almost entirely leaves. Phytochemical tests on samples consisted of steroid, flavonoid, phenol, saponin, and triterpenoid tests.