Fresh Leaf Extract Research Articles

SYNTHESIS OF SILVER NANOPARTICLES USING AQUEOUS EXTRACT OF PERGULARIA DAEMIA AND ANALYSIS OF ANTIMICROBIAL ACTIVITY

The present work deals with the green synthesis of silver nanoparticle from aqueous extract of Pergularia daemia as reducing agent and evaluation of the antimicrobial potential of synthesized green nanoparticles (GNPs). The synthesized silver nanoparticles (SNPs) were characterized by UltravioletVisible absorption spectroscopy (UV-Vis) and high-resonance transmission electron microscopy (TEM) analysis. Visual observation showed that the color of the fresh leaf extracts of P. daemia turned into dark brown after incubation of 24 h with Ag precursors. The TEM analysis showed that nanoparticles were spherical in shape and the size was found to be in the range of 7-22 nm. The green synthesized nanoparticles showed concentration dependent (25 µg/mL, 50 µg/mL and 100 µg/mL) noteworthy antimicrobial activity against E. coli, S. aureus and B. subtilis with ciprofloxacin as a standard. Research findings conclude that GNPs possess superior antimicrobial potential and it is a new option to combat antibiotic resistance.

A rapid method to assess the formaldehyde dehydrogenase activity in plants for the remediation of formaldehyde.

The formaldehyde dehydrogenase (FADH) activity in plants is essential to the removal of airborne formaldehyde (FA) by plants. A rapid and efficient method was established to assess the FADH activity in plants by analyzing the efficiencies of the extracts of fresh and enzyme-inactivated leaves to degrade FA, with the enzyme-inactivated leaves prepared by freezing with liquid nitrogen. The efficiencies of airborne FA dissipated by different plants were evaluated through the FA fumigation experiments using four selected plants, with the results analyzed against the calculated leaf FADH activities. Fresh and enzyme-inactivated leaf extracts degraded FA to different extents. The degradative efficiencies of leaf extracts were positively related to the initial FA test levels at 6-18mgl-1. The relative plant-leaf FADH activities formed the order of Chenopodium album L. > Atenia cordifolia > Plantain > Aloe, which was in line with the observed FA dissipating efficiencies of the plants exposed to 0.72mgm-3 airborne FA for 24h. Other dominant degrading mechanisms in plant leaves resulted in higher dissipating efficiencies of Plantain over that of Atenia cordifolia when exposed to 1.56mgm-3 FA for 24h. The established method could be applied to estimate the FADH activity in plants for assessment of the plant remediation efficiency of FA in air at lower concentrations.

Potency of Rosela Leaves (Hibiscus sabdariffa L.) as a Hepatoprotector to Histological Structure Damage of the Hepar of Mice Male (Mus musculus L.) Strain DDW that Proposed Natrium Nitrite (NaNO2)

The study of Rosela Leaf Extract As A Protective Agent Against The Damage Of Liver Tissue OfMale Mice Strain DDW that has been Induced By Sodium Nitrite was conducted. The experiment used Completely Randomized Design. The treatment given was three different dosages of rosella leaf extract, namely 14 mg / ml, 28 mg / ml and 42 mg / ml and without any extract as a control. All mice have been induced by sodium nitrite for 18 days followed by healing process through injection of 0.3 ml of rosella leaf extract at different concentration per day during 14 days. The result showed that he control, treatment of 14 mg / ml and 42 mg / ml affected the color change of liver tissue. However, the control increased liver tissue weight compared to all treatments. The damage levels of hepatocyte tissue treated by all different dosages of fresh rosella leaf extract decreased compared to the damage level of control. It could be concluded that fresh rosella leaf extract showed potential agent to protect liver mice from damage caused by sodium nitrite induction.

Synthesis and Characterization of Silver Nanoparticles from <I>Ashyranthus aspera</I> Extract for Antimicrobial Activity Studies

Development of biologically inspired experimental processes for the synthesis of nanoparticles is evolving into an important branch of nanotechnology. Plant-mediated synthesis of nanomaterials has been increasingly gaining popularity due to its eco-friendly nature and cost-effectiveness. In the present study, we were synthesized silver (Ag) nanoparticles using aqueous extracts of fresh leaves of Ashyranthus aspera medicinal plants as bio-reducing agents. UV-Vis spectrometer used to monitor the reduction of Ag ions and the formation of AgNPs in the medium. UV-Vis spectra and visual observation showed that the color of the fresh leaf extracts of Ashyranthus aspera turned into grayish-brown respectively, after treatment with Ag precursors. XRD and SEM have been used to investigate the morphology of prepared AgNPs. The peaks in the XRD pattern are associated with that of the Face-Centered-Cubic (FCC) form of metallic silver. TGA/DTA results associated with weight loss and exothermic reaction due to the desorption of chemisorbed water. FTIR was performed to identify the functional groups which form a layer covering AgNPs and stabilize the AgNPs in the medium. Moreover, silver nanoparticles using aqueous leaf extracts of Ashyranthus aspera were separately tested for their antibacterial activity against Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Enterobacter). The results showed that the bacterial growth was inhibited by the extracts containing AgNPs Nanoparticles. The biosynthesized nanoparticle was prepared from Ashyranthus aspera leaf extracts exhibits potential applications as broad-spectrum antimicrobial agents
 Keywords: Ashyranthus aspera, Silver Nanoparticles, Plant extracts, Bacteria, Antibacterial activity.

Germination and Early Seedling Growth of Two Rice Varieties as Affected by Invasive Ageratina adenophora

Ageratina adenophora is a perennial herb of family Asteraceae expanding as a serious threat to ecological integrity and biodiversity. This study aims to assess the allelopathic potential of fresh and dry leaf and root aqueous extracts of this invasive species on germination, early seedling growth, seedling biomass and seed vigor of two rice (Oryza sativa L.) varieties namely 6444 and 2245. The result revealed that with increasing concentration of extract, inhibition effect also increases. Leaf extract was more inhibitory than root extract and dry plant extracts had higher allelopathic impact in comparison to fresh plant extracts. Among the two investigated rice varieties 6444 was more resistant to the fresh and dry leaf extract while 2245 was more resistant to fresh and dry root extract. This study will be helpful in assessing the effect caused by A. adenophora to rice crop and could be used in devising weed control strategy.

FORMULATION AND EVALUATION OF ANTI-ACNE GEL CONTAINING MURRAYA KOEINIGII EXTRACT

Objective: Acne, one among the very fashionable socially distressing skin conditions created by Propionibacterium acne have generally been treated by antibiotics. Within the light of the growing threat of antibiotic resistance, natural plant products are applied as a safer alternative. Keeping the very fact in the background, during this research work, the formulation of gel from the extracts of Murraya koeinigii leaves are prepared and evaluated as an anti-acne drug.
 Methods: The fresh leaf extracts were subjected to phytochemical and antimicrobial screening. Minimum Inhibitory Concentration (MIC) decided. Gel formulation of the extracts was developed and evaluated. The manufactured formulations were subjected to In vitro antibacterial activity against P. acnes, S. epidermidis and S. aureus. The marker compound, clindamycin, in herbal anti-acne preparation, was kept for the comparison with the zones of inhibition for antibacterial activity.
 Results: Anti-acne property was explored with the help of a standard curve and by comparing diffusion profiles by taking clindamycin as a reference.
 Conclusion: From the present study it can be concluded that addition of permeation enhancer in the test formulation will improve the diffusion profile and thus it was designed to add permeation enhancer.

Allelopathic Interference of Ageratum conyzoides L. against Rice Varieties

Ageratum conyzoides L. is an invasive weed that has severely infested cultivated lands and interferes with the growth of crops. In this study aqueous extracts of 50 and 100% concentration were prepared from fresh and air dried plant parts (Leaves and roots) of A. conyzoides and their allelopathic effect was observed on seed germination and seedling growth of two rice varieties namely Sava and Geru. In Sava variety, a significant reduction in seed germination, seedling length and dry weight was recorded under dry leaf extract while under fresh leaf extract seed germination and seedling length was increased at 50% concentration as compared to control. In Geru variety both fresh and dry leaf extracts resulted in decreased seed germination and seedling growth with increasing concentration. Similar patterns were recorded for fresh and dry root extracts. The inhibitory effect of leaf extracts was more pronounced than root extract and overall dry aqueous extracts of leaf and root were more inhibitory than fresh aqueous extracts. Among varieties, Sava was least affected as compared to the Geru. This study indicated that A. conyzoides exert allelopathic effect towards rice crop by releasing water soluble phytochemicals.

Phytochemical Screening and Allelopathic Effects of Ageratum conyzoides L.

A weed is a plant that grows and compete with other plants for food, light, moisture and space. Ageratum conyzoides L. is a common weed belonging to the Asteraceae family. They are annual aromatic weed. They are invasive of pastures, vacant lots and even entire forest areas. Phytochemicals are secondary metabolites that are known to influence various activities of plants. The qualitative phytochemical analysis of the methanol leaf extract of A. conyzoides revealed the presence of phenols, flavonoids, alkaloids, quinones, cardiac glycosides, terpenoids and tannins except saponins and anthraquinones, whereas the quantification revealed that the methanol extract contained flavonoid content (27.8±0.685 mg/g), phenol content (8.6±0.446 mg/g) and alkaloid content (2.06±0.53 mg/g). The fresh leaf extract of A. conyzoides was treated to the seeds of Zea mays to study the allelopathic effect of the weed. The germination rate shows a gradual decline with increase in the concentration of the weed extracts.

Use of mangrove leaves (Rhizophora mucronath) extract as a biofertilizer for seed germination

The aqueous fresh leaf extract of Rhizophora mucronata Lam. is used for germination of Triticum aestivum (Wheat) and Brassica juncea (Mustard) seeds. The seeds treated with the aqueous extract showed high percentage of germination as well as longer root and shoot length as compared to control and other commercially used biofertilizer like Sargassum treated seeds.

English

The jackfruit tree (Artocarpus heterophyllus Lam.) is considered one of the most versatile trees and is used from wood production to traditional medicine. In traditional medicine, leaves are used to treat asthma, wounds and abscesses, and prevent ringworm infection. Several biological activities have been documented for jackfruit extracts confirming its popular use; however, its in vitro cytotoxicity and anti-herpesvirus activity are rarely discussed. The objective of this study was to explore the in vitro anti-herpesviral potential of A. heterophyllus leaf extracts at non-cytotoxic levels. Leaves were obtained from specimens located in the Atlantic forest biome. Dried and fresh ethanolic, methanolic and hexanic leaf extracts were obtained from the partition of a crude ethanolic extract. The cytotoxicity was performed on erythrocyte and kidney cell lines. The antiviral test was against equine, suid, and bovine herpesviruses. The contents of phenolic compounds were established from methanolic leaf extracts. Fresh leaf extracts did not show toxicity to erythrocytes in any concentration tested; however, dry leaf extracts showed >10% of hemolysis at ≥200 µg/ml. The maximum non-cytotoxic concentrations for Vero and MDBK cells ranged from 7.8 to 125 µg/ml. The content of phenolic compounds was 22.02% higher in fresh leaf than in dry leaf methanolic extracts. Methanolic extracts showed antiviral activity against BoHV-1 (PI of 99.20%) and SuHV-1 (PI of 94.38%). A. heterophyllus fresh and dry leaf extracts are remarkable as anti-herpesviruses and candidates to proceed for in vivo tests. Key words: Artocarpus heterophyllus Lam., Jackfruit tree, antiviral activity, kidney cell-line, erythrocyte, medicinal plant.

Antibacterial Activity Test of Wet and Dried Extracts of Calabash Tree (Crescentia cujete L.) against Aeromonas hydrophilla

Study to determine the antibacterial activity of wet and dry extract of the leaf, fruit, and bark of Calabash tree (Crescentia cujete L.) against the growth of Aeromonas hydrophila. The solvent extraction process was done by using 96% ethanol in the maceration method. Antibacterial test results using diffusion agar to decide clear zone and tube series of dilution test to provide MIC and MBC. Fresh leaf extract produces the highest clear zone diameter (20.06 mm), after which fresh bark extract (12.81 mm), and the last is fresh fruit extract (3.22 mm). In contrast to fresh extracts, the dried extracts are have not clear zone. MIC (Minimum Inhibitory Concentration) of Calabash Tree fresh leaf extract against Aeromonas hydrophila is 80%, and MBC (Minimum Bactericidal Concentration) is 100%.

ANTIBACTERIAL ACTIVITY OF HERBAL EXTRACTS AGAINST Vibrio parahaemolyticus CAUSING ACUTE HEPATOPANCREATIC NECROSIS DISEASE (AHPND) IN WHITELEG SHRIMP (Litopenaeus vannamei) IN VITRO

The antibacterial activity of herbal extracts to Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease in whiteleg shrimp (Litopenaeus vannamei) was studied and carried out in the School of Agriculture and Aquaculture, Tra Vinh University. The results showed that the inhibition zone of Muntingia calaburaleaf extract with ethanol 70% was 18.00 ± 0.00mm, Muntingia calabura leaf extract with ethanol 90% was 17.33 ± 0.58 mm and Muntingia calabura fresh leaf extract was 13.00 ± 0.00 mm. Minimum inhibitory concentration (MIC) of Muntingia calabura leaf extract with ethanol 70% and 90% were 5.120 mg/L, 10.240 mg/L. MIC value of Al lium cepa extract with ethanol 70% and 90% were 40.960 mg/L, Physalis angulata extract with ethanol 70% and 90% were 81.920 mg/L. The results showed that the antimicrobial activity was highest in the Muntingiacalabura leaf extract and lowest in Physalis angulata extract.

Alteration in the oxidative status of Drosophila melanogaster Meigen (Diptera: Drosophilidae) fed with a diet containing Centaurea depressa M. Bieb. (Asteraceae)

Abstract The aim of the this study was to evaluate the effects of fresh, dried and freeze-dried Centaurea depressa M. Bieb. (Asteraceae) on the oxidant and antioxidant status of the model organism D. melanogaster Meigen (Diptera: Drosophilidae) experimentally. The study was carried out from 2016 to 2019, and plant leaf extracts (0-50 mg/l) were added to insect standard artificial diets. The total protein, protein carbonyl content and glutathione-S-transferase, superoxide dismutase and catalase activities were quantified at the insect’s third larval stage. Our data showed that protein carbonyl content varied from 2.70 nmol/mg protein in the control group to 59.11 nmol/mg protein in the group fed with fresh leaf extract signifying induction of oxidative stress. All extracts increased the levels of all antioxidant enzymes and decreased the amounts of total protein. Meanwhile, the group fed with the freeze-dried extract showed no significant difference in the levels of total protein and protein carbonyl content except at the 50 mg/l concentration of the extract. Moreover, this group had superoxide dismutase and catalase activities 4 to 5 times higher than in the control group. In conclusion, induction of oxidative stress indicates that the fresh form of C. depressa leaves may have potential as a natural pesticide, whereas induction of endogenous antioxidant enzymes by the freeze-dried extract suggest its potential as an antioxidant.

Antibacterial, antibiofilm activity and cytotoxicity of crude extracts of Ptaeroxylon obliquum (Ptaeroxylaceae) used in South African ethnoveterinary medicine against Bacillus anthracis Sterne vaccine strain

Anthrax, a zoonotic disease and a potential biological weapon for terrorism is caused by Bacillus anthracis, a Gram‐positive, sporulating bacterium. Anthrax is endemic in South Africa and primarily affects livestock and wildlife species. Problems with the control of anthrax include growing resistance to present antibiotics and disinfectants or decontaminants coupled with low antibiotic access in rural and poor settings. Therefore, it is imperative to search for new antimicrobials, especially from natural sources that could offer effective and potent alternatives. Medicinal plants are putative sources of new antimicrobials. Ptaeroxylon obliquum (Ptaeroxylaceae) is used traditionally in South Africa as an anthrax remedy in livestock. Here, the antibacterial and antibiofilm activity as well as cytotoxicity of the bark and fresh leaf crude extracts of P. obliquum was determined.A two‐fold serial microdilution assay was used to determine the minimum inhibitory concentration (MIC) of the acetone and chloroform extracts of bark and the chloroform extract of fresh leaves of the plant against B. anthracis Sterne vaccine strain. The crystal violet assay was used to determine antibiofilm activity while cytotoxicity was determined using a tetrazolium‐based colorimetric (MTT) assay against Caco‐2 intestinal cells.The MIC of the extracts ranged from 0.005 – 0.039 mg/mL compared to the positive control gentamicin (MIC = 4.9 × 10−5) with acetone bark extract being the most active (MIC = 0.005 mg/mL). Activity of extracts against B. anthracis biofilms was high (>50 % antibiofilm activity) at 0 h (prevention of biofilms; ranged between 87–99 %), and 24 h (destruction of pre‐formed biofilms; ranged between 58–93 %). The chloroform fresh leaf extract had the best antibiofilm activity. The acetone bark extract had the best total antibacterial activity (yield (mg/g)/MIC (mg/mL)) of 21 500 g/mL. This means that the mass extracted from 1 g of the acetone extract can be diluted by 21 500 times and still inhibit bacterial growth. The 50% lethal concentration (LC50) of the extracts against Caco‐2 cells ranged between 0.03–0.19 mg/mL with the chloroform fresh leaf extract being relatively the safest (LC50 = 0.19 mg/mL). The acetone bark extract had the best selectivity index (LC50/MIC) of 6. This shows that the extract has higher toxicity against the bacteria than to mammalian cells.The results support the use of the plant against anthrax in South African ethnoveterinary medicine. The extracts may be potential alternatives to chemicals as disinfectants or decontaminants, especially in rural areas where chemicals are not accessible to the farmers. The compounds responsible for anti‐B. anthracis activity, and their mechanism(s) of action, is the focus of future studies.Support or Funding InformationThe University of Pretoria is thanked for providing research support

Green Synthesis, characterization of Zinc oxide nanoparticles using Chromolaena odorata extract and evaluation of its properties for photoanode of solar cells

Synthesis of Zinc oxide nanoparticles (ZnONPs) using plant extract was carried out in this work because of its advantages over other method of synthesis namely; simplicity, environmental friendliness and elimination of toxic chemicals. Fresh Siam leaf extract was characterized over a period of 10 days to study the effect of time on the optical photon energy absorption properties. ZnONPs was synthesized and deposited on substrate by spin-coating method and characterized by UV-vis spectroscopy, Fourier transform infra-red spectroscopy, Scanning Electron microscopy, and Energy Dispersive X-ray. ZnONPs thin film device was then fabricated to study the electrical properties. From UV-vis spectroscopic result, as the time (day) increased, there was an increase in the value of the transmittance and corresponding reduction in photon energy absorption. FTIR result gives the functional groups and absorption number at 3448.04cm−1 O-H single bond, 2524.85cm−1 C-H bond, 1437.15 cm−1, 880.59 cm−1, 729.06 cm−1, 433.44 cm−1 Zn-OH and Zn-O variety of single bond. The surface morphology shows large grain size. Energy band gap of ZnONPs was approximately 3.7 eV. The fabricated ZnONPs thin film device under illumination has efficiency of 2.010/0. From the morphology, optical, and electrical properties of the ZnONPs thin film and device, it could be a suitable material for crystalline solar cells as photoanode.

Biochemical Analyses of Male Rat’s Serum Administered Dexamethasone and Green Synthesis Cerium Oxide Nanoparticles Treatment

Current research involves preparing cerium oxide nanoparticles using simple, environmental- friendly green synthesis approach via fresh green tea leaf extract (Camellia sinensis) as a capping and reducing agent, Ultraviolet-visible spectroscopy (UV-Vis.) is used to check the formation of nanoparticles by investigate the absorption peak and energy gap (Eg); Maximum absorption peak was around 295nm and Eg was 4.2eV calculated using Planck’s equation, X-Ray diffraction (XRD) analysis of Nano powder stated that Cerium Oxide has a cubic fluorite structure with face centered cubic (FCC) structure, blood serum glucose levels, liver functions after CeO2 Nanoparticles treatment for rats were reported in this paper. Forty eight adult male rats were used and divided randomly into six groups each group has eight rats (n=8), Results stated that blood glucose levels, liver enzymes were increased in groups that injected by dexamethasone while considerably modified in rats administered by Cerium Oxide Nanoparticles.

Profiling and structural analysis of cardenolides in two species of Digitalis using liquid chromatography coupled with high-resolution mass spectrometry

Plants of the Digitalis genus contain a cocktail of cardenolides commonly prescribed to treat heart failure. Cardenolides in Digitalis extracts have been conventionally quantified by high-performance liquid chromatography yet the lack of structural information compounded with possible co-eluents renders this method insufficient for analyzing cardenolides in plants. The goal of this work is to structurally characterize cardiac glycosides in fresh-leaf extracts using liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) that provides measured accurate mass. Fragmentation of cardenolides is featured by sequential loss of sugar units while the steroid aglycone moieties undergo stepwise elimination of hydroxyl groups, which distinguishes different aglycones. Using a reverse-phase LC column, the sequence of elution follows: diginatigenin→digoxigenin→gitoxigenin→gitaloxigenin→digitoxigenin for cardenolides with the same sugar units but different aglycones. A linear range of 0.8-500 ng ml−1 has been achieved for digoxigenin, β-acetyldigoxin, and digitoxigenin with limits of detection ranging from 0.09 to 0.45 ngml−1. A total of seventeen cardenolides have been detected with lanatoside A, C, and E as major cardenolides in Digitalis lanata while seven have been found in Digitalis purpurea including purpurea glycoside A, B, and E. Surprisingly, glucodigifucoside in D. lanata and verodoxin and digitoxigenin fucoside in D. purpurea have also been found as major cardenolides. As the first MS/MS-based method developed for analyzing cardenolides in plant extracts, this method serves as a foundation for complete identification and accurate quantification of cardiac glycosides, a necessary step towards understanding the biosynthesis of cardenolide in plants.

Green synthesis, characterization and photocatalytic applications of silver nanoparticles using Diospyros lotus

AbstractCurrently, the green route for synthesis of silver nanoparticles (Ag NPs) using plants leaf extract is an emerging research area in nanotechnology. The present study was explored for synthesis, characterization and catalytic application of Ag NPs using Diospyros lotus fresh leaf extracts. Factors affecting the synthesis were optimized and optimum conditions were pH of 8.6, silver nitrate (AgNO3) concentration of 1.5 mM and 10 mL leaf extract. Formation of Ag NPs was observed by change in color of reaction mixture from pale yellow to reddish brown. The synthesized Ag NPs were characterized using UV-Vis spectrophotometer, EDX, XRD and SEM analyses. UV-Vis spectrophotometer showed maximum absorbance peak in the range of 407 nm at different time intervals indicating formation of Ag NPs. SEM and XRD analysis confirmed face centered cubic structure and crystalline nature of biologically synthesized Ag NPs with average particle size of 27 nm. The purity of synthesized Ag NPs was revealed by EDX. Finally, photo catalytic activity (PCA) of Ag NPs was studied and 72.91% decolorization of industrial waste water was obtained at 54 h. Some important parameters like pH, turbidity, conductance; TSS, TDS, sulphide, sulphates, etc. were also monitored before and after treatment with Ag NPs.

Fresh leaf extract’s efficacy of twelve medicinal plants against Culex quinquefasciatus mosquito larvae

Fresh leaf extract’s efficacy of twelve medicinal plants against Culex quinquefasciatus mosquito larvae

Comparative Analyses of the Efficacy of Dry and Fresh <i>Mangifera indica</i> Ethanolic Extracts on <i>E. coli</i> and <i>S. aureus</i>

Background: The need to identify and characterize new antimicrobial agents is important due to the increasing development of resistance by microorganisms to the existing antimicrobial agents. Aim: This study examined the efficacies of Mangifera indica on Escherichia coli and Staphylococcus aureus. Method: Three parts (leaf [L], root [R], and bark [B]) of the plant were analyzed. The extraction of the samples was performed by aseptically grinding the samples, dissolving in absolute ethanol, and filtering through whatman filter paper. The efficacy of the extracts bothsingle and combined was determined using agar well diffusion assay with gentamycin [10 μl] (E. coli) and vancomycin [30 μl] (S. aureus) as control antibiotics. Results: The higher concentration (C2 = 3.0 g/ml) showed more antibacterial effectiveness than the lower concentration (C1 = 1.5 g/ml) against both bacterial isolates with significant differences (P E. coli dry leaf extract; fresh bark extract), double extracts (S. aureus: dry and fresh leaf extracts) and triple extract (E. coli and S. aureus dry extracts). For the single extracts the bacteria has the following significant results: E. coli L (dry 6.3 ± 2.5 mm, fresh 14.7 ± 0.6 mm, P = 0.0050), R (dry 11.3 ± 1.5 mm, fresh 7.3 ± 1.5 mm, P = 0.0327); for S. aureus L (dry 7.0 ± 1.7 mm, fresh 11.0 ± 1.0 mm, P = 0.0257), R (dry 7.0 ± 2.0 mm, fresh 11.7 ± 1.5 mm, P = 0.0325), and B (dry 5.0 ± 1.0 mm, fresh 16.0 ± 1.0 mm, P = 0.0002). For the double extracts the bacteria has the following significant results: E. coli L + R (dry 15.7 ± 2.3 mm, fresh 1.7 ± 1.5 mm, P = 0.0070), R + B (dry 18.7 ± 1.5 mm, fresh 9.7 ± 1.5 mm, P = 0.0020), and L + B (dry 9.7 ± 1.5 mm, fresh 6.3 ± 0.6 mm, P = 0.0241); S. aureus L + R (dry 14.7 ± 1.5 mm, fresh 7.0 ± 1.0 mm, P = 0.0019), R + B (dry 15.3 ± 1.5 mm, fresh 11.7 ± 1.5 mm, P = 0.0424). For the triple extracts, the fresh leaves showed significantly higher levels of efficacy than the dry for both E. coli L + R + B (P = 0.0101) and S. aureus (P = 0.0307). The fresh extracts showed higher levels of efficacy than dry extracts against both bacteria for all the single and three combined conditions. Conclusions: Fresh extracts show better efficacies against E. coli while dry extracts show greater efficacies against S. aureus for both single and triple combined extracts. The reverse is true for double combined extracts.