scholarly journals Antibacterial Activities of Echinops kebericho Mesfin Tuber Extracts and Isolation of the Most Active Compound, Dehydrocostus Lactone

2021 ◽  
Vol 11 ◽  
Author(s):  
Serawit Deyno ◽  
Andrew G. Mtewa ◽  
Derick Hope ◽  
Joel Bazira ◽  
Eyasu Makonnen ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Essential Oil ◽  
Spectroscopic Method ◽  
Ethanol Extract ◽  
Antibacterial Activities ◽  
Dilution Method ◽  
Chloroform Fraction ◽  
Isolation And Purification ◽  
E Coli ◽  
Dehydrocostus Lactone

Echinops kebericho Mesfin is traditionally used for the treatment of various infectious diseases. This study investigated antibacterial activity of the essential oil (EO) and the different fractions of ethanol extract. The most active component was isolated and identified. Isolation and purification was accomplished using chromatographic techniques while identification was done by spectroscopic method. Minimum inhibitory concentration (MIC) was determined using the broth micro-dilution method. In bioactive-guided isolation, percent inhibition was determined using optical density (OD) measurement. The MICs of the essential oil ranged from 78.125 μg/ml to 625 μg/ml, and its activity was observed against methicillin-resistant Staphylococcus aureus (MRSA, NCTC 12493). Ethyl acetate fraction showed high activity against MRSA (NCTC 12493), MIC = 39.075 μg/ml followed by Enterococcus faecalis (ATCC 49532), MIC = 78.125 μg/ml and was least active against Klebsiella pneumoniae (ATCC 700603), MIC = 1,250 μg/ml. MIC of hexane fraction ranged from 156.2 µg/ml to Escherichia coli (ATCC 49532) to 1,250 μg/ml to E. coli (NCTC 11954). The MICs of chloroform fraction ranged from 312.5 to 2500 μg/ml; while butanol fraction could be considered pharmacologically inactive as its MIC value was 2,500 μg/ml for all and no activity against E. coli (NCTC 11954). Dehydrocostus lactone was successfully isolated and identified whose MIC was 19.53 μg/ml against MRSA. Dehydrocostus lactone isolated from E. kebericho M. showed noteworthy antibacterial activity which lends support to ethnopharmacological use of the plant. Further optimization should be done to improve its antibacterial activities and pharmacokinetic profile.

scholarly journals Antibacterial Activities of Essential Oil and Fractions of Ethanolic extract of Echinops kebericho Tuber

2020 ◽  
Author(s):  
Serawit Deyno ◽  
Derick Hope ◽  
Joel PhD Ba ◽  
Eyasu Makonnen ◽  
Paul E. Alele
Keyword(s):  
Antibacterial Activity ◽  
Essential Oil ◽  
Ethyl Acetate ◽  
Ethanolic Extract ◽  
Ethyl Acetate Fraction ◽  
Antibacterial Activities ◽  
Assay Method ◽  
Chloroform Fraction ◽  
E Coli ◽  
Routes Of Administration

Abstract Background: Echinops kebericho is an endemic medicinal plant in Ethiopia traditionally used for treatment of various infectious diseases through different routes of administration such as inhalation, orally chewed, and topically sprayed to affected area. This study investigated antibacterial activity of the essential oil (EO) and the different fractions of ethanolic extract of Echinops kebericho tuber.Results: MIC of EO ranged from 78.125 µg/ml – 625 µg/ml and it was most active against methicillin resistant S. aureus (MRSA, NCTC 12493). Ethyl acetate fraction showed better activity against MRSA (NCTC 12493), MIC = 39.075 µg/ml followed by E. faecalis (ATCC 49532), MIC = 78.125 µg/ml and was least active against K. pneumonia (ATCC 700603), MIC = 1250 µg/ml. MIC of hexane fraction ranged from 156.2- µg/ml to E. faecalis (ATCC 49532) to 1250 µg/ml to E. coli (NCTC 11954). Chloroform fraction MIC ranged from 312.5 - 2500µg/ml; while butanol fraction could be considered pharmacologically inactive as its MIC value is 2500 µg/ml for all and no activity shown for E. coli (NCTC 11954). The MIC of DMSO against all strains ranged 12.5- 25 % w/v. The MIC estimated by OD measurement correlated well with that of resazurin assay method.Conclusion: Essential oil and ethyl acetate fraction showed noteworthy antibacterial activity, and MRSA was the most susceptible strain. Further study, however, should be done to investigate the effect on the isolated active component(s).

scholarly journals Aktivitas Antibakteri Sabun Cuci Tangan yang Mengandung Ekstrak Metanol Rumput Laut Eucheuma spinosum (Antibacterial Activity Test of Eucheuma spinosum Methanol Extract Hand Wash)

MEDULA ◽  
2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Nur Illiyyin Akib ◽  
Mariani Triwatami ◽  
Andi Eka Purnama Putri
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Antibacterial Activity ◽  
Methanol Extract ◽  
Antibacterial Activities ◽  
Dilution Method ◽  
E Coli ◽  
Hand Wash ◽  
Ethanol Extracts ◽  
Physical And Chemical

Background: Eucheuma spinosum seaweed contain flavonoid, triterpenoid, alkaloids and polyphenol which has been widely used in antibacterial activity. Purpose(s):The aim of the research are to determine antibacterial activities of metanol extracts of E. spinosum against Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922; to formulate hand wash of seaweed of methanol extract E. spinosum that has antibacterial activity and has physical and chemical stability. Methods: Seaweed E.spinosum methanol extract was derived by maceration method. Antibacterial activities of the extract were tested by liquid dilution and solid dilution method. Hand wash was formulated by mechanical dissolved methods. Antibacterial activity of hand wash were tested by liquid dilution and solid dilution method. Physical and chemical stabilities were conducted by cycling test. Results:. These were showed through minimum inhibitory concentration (MIC) of ethanol extracts of E. spinosum against S. aureus ATCC 25923 at concentrations of 6% and E. coli ATCC 25922 at concentrations of 6%. The minimum bactericidal concentration (MBC) of ethanol extracts of E. spinosum against S. aureus ATCC 25923 at concentrations of 8% and E. coli ATCC 25922 at concentrations of 8%. Formulation of hand wash seaweed E. spinosum methanol extract at concentrations of 8% and 10%. The test of stabilities results of hand wash changes were organoleptic, viscosity, pH, and foaming ability were accordance to qualified standard. The antibacterial activity of hand wash contains seaweed E. spinosum methanol extract at concentration of  8% and 10% have bactericidal activity againts S. aureus ATCC 25923 and E. coli ATCC 25922. Conclusion: Overall, these results suggested that formula of hand wash contains E. spinosum metanol extracts have antibacterial properties against S. aureus ATCC 25923 and E. coli ATCC 25922.Keywords: antibacterial, Eucheuma spinosum, hand wash, physical and chemical stability Latar Belakang: Rumput laut Eucheuma spinosum mengandung flavonoid, triterpenoid, alkaloid, dan polifenol yang memiliki aktivitas antibakteri. Tujuan: Penelitian ini bertujuan mengetahui aktivitas antibakteri ekstrak metanol E. spinosum terhadap Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922; membuat sediaan sabun cuci tangan dari ekstrak metanol E. spinosum yang memiliki aktivitas antibakteri dan stabil secara fisika dan kimia. Metode: Ekstrak metanol E. spinosum diperoleh dengan metode maserasi. Uji aktivitas antibakteri ekstrak dilakukan dengan metode dilusi cair dan dilusi padat. Sabun cuci tangan diformulasi dengan metode pencampuran mekanik. Uji aktivitas antibakteri sabun cuci tangan dilakukan dengan metode dilusi cair dan dilusi padat. Uji stabilitas fisika kimia dilakukan dengan metodecycling test. Hasil: Konsentrsi Hambat Minimum (KHM) ekstrak metanol E. spinosum terhadap S. aureus ATCC 25923 adalah 6% dan terhadap E. coli ATCC 25922 adalah 8%. Ekstrak metanol E. spinosum dapat diformulasi menjadi sabun cuci tangan dengan konsentrasi 8% dan 10%. Uji stabilitas menunjukkan bahwa perubahan organoleptik, viskositas, pH, dan kemampuan membentuk busa masih berada dalam nilai yang dipersyaratkan. Uji aktivitas antibakteri sabun cuci tangan yang mengandung ekstrak metanol E. spinosum dengan konsentrasi 8% dan 10% memiliki aktivitas antibakteri terhadap ATCC 25923 and E. coli ATCC 25922. Simpulan: Formula sabu cuci tangan yang mengandung ekstrak metanol E. spinosum memiliki aktivitas antibakteri terhadap ATCC 25923 and E. coli ATCC 25922.Kata kunci: antibakteri, Eucheuma spinosum, sabun cuci tangan, stabilitas fisika kimia

scholarly journals Anti-HIV, Antitubercular and Antibacterial Activities of Novel 3-(Substituted Quinazolinylamino)-2-phenyl quinazolin-4(3H)ones

2019 ◽  
Vol 32 (2) ◽  
pp. 281-286
Author(s):  
M.T. Sulthana ◽  
K. Chitra ◽  
V. Alagarsamy
Keyword(s):  
Antibacterial Activity ◽  
Antitubercular Activity ◽  
Antibacterial Activities ◽  
Primary Amines ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
E Coli ◽  
Agar Dilution ◽  
Anti Hiv Agents ◽  
Anti Hiv

In the present study, we have synthesized a series of novel 2-phenyl-3-(substituted quinazolinylamino)quinazolin-4(3H)-ones by the reaction of 3-(substituted)-2-hydrazinoquinazoline-4(3H)-ones with 2-phenyl-3,1-benzoxazin-4-one. The starting material 3-(substituted)-2-hydrazinoquinazolin-4(3H)-ones were synthesized from various primary amines. All the synthesized compounds were screened for their antitubercular, anti-HIV and antibacterial activity against different Grampositive and Gram-negative strains by agar dilution method. Among the test compounds, 3-(4-nitrophenyl)-2-(4-oxo-2-phenylquinazolin-3(4H)-ylamino)quinazolin-4(3H)-one (BQZ6) and 3-(4-chlorophenyl)-2-(4-oxo-2-phenylquinazolin-3(4H)-ylamino)quinazolin-4(3H)-one (BQZ7) shown most potent antibacterial activity against E. coli, P. aeruginosa and S. aureus with the MIC of 3 μg/mL. The compound BQZ7 exhibited the antitubercular activity with the MIC of 25 μg/mL and anti-HIV activity with the MIC of 35.4 μg/mL against HIV1 and HIV2 and offers potential lead for further optimization and development to new antitubercular and anti-HIV agents. The results from this study confirm that the synthesized and biologically evaluated quinazolines showed promising antimicrobial, antitubercular and anti-HIV activities and are new scaffolds for antimicrobial activity

scholarly journals Antibacterial Activity and Mechanism of Litsea cubeba L. Essential Oil Against Acinetobacter baumannii

2021 ◽  
Vol 16 (3) ◽  
pp. 1934578X2199914
Author(s):  
Kaiyuan Hao ◽  
Baichang Xu ◽  
Geyin Zhang ◽  
Feifei Lv ◽  
Yuhan Wang ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Essential Oil ◽  
Cell Membrane ◽  
Acinetobacter Baumannii ◽  
Growth Curves ◽  
Antibacterial Activities ◽  
Dilution Method ◽  
Obvious Effect ◽  
Strong Activity ◽  
Litsea Cubeba

The main objective of this study was to investigate the antibacterial activity and mechanism of Litsea cubeba essential oil (LCEO) against Acinetobacter baumannii. The antibacterial activity was examined by a serial dilution method and growth curves. The essential oil showed strong activity against A. baumannii. The mechanism of the antibacterial action was evaluated by the integrity and permeability of the membrane, scanning electron microscopy ( SEM) and SDS-PAGE. The change in permeability of the cell membrane and leakage of cell intracellular biomacromolecules verified that LCEO has an obvious effect on the cell membrane. SEM showed the damaging effect of the essential oil on cells since the morphology of the treated bacteria was significantly changed. Different expression of proteins indicated the effect of the essential oil on protein synthesis. It is suggested that LCEO with both emulsifying and antibacterial activities, has a potential to serve as an inhibitor against A. baumannii.

scholarly journals Actividad antibacteriana in vitro de Curcuma longa (Zingiberaceae) frente a bacterias nosocomiales en Montería, Colombia

2016 ◽  
Vol 64 (3) ◽  
Author(s):  
Nelson Méndez Álvarez ◽  
Alberto Angulo Ortíz ◽  
Orfa Contreras Martínez
Keyword(s):  
Antibacterial Activity ◽  
Essential Oil ◽  
Clinical Isolate ◽  
Curcuma Longa ◽  
Antibacterial Properties ◽  
Ethanolic Extract ◽  
Ethanol Extract ◽  
Future Research ◽  
E Coli

Bacterial resistance is a growing health problem worldwide that has serious economic and social impacts, compromising public health, and the therapeutic action of current antibiotics. Therefore, the search for new compounds with antimicrobial properties is relevant in modern studies, particularly against bacteria of clinical interest. In the present study, in vitro antibacterial activity of the ethanol extract and essential oil of Curcuma longa (Zingiberaceae) was evaluated against nosocomial bacteria, using the microdilution method. Escherichia coli strains, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus sp. were used, Salmonella sp. and Bacillus sp., isolated from nosocomial infections in a hospital in the city of Monteria and reference strains of S. aureus ATCC 43300, S. aureus ATCC 29213, S. aureus ATCC 25923, P. aeruginosa ATCC 27853, E. coli ATCC 25922 and K. pneumonia ATCC 700603. The ethanol extract antibacterial profile was more efficient at higher concentrations (1 000 ppm), obtaining significant percentages of reduction of more than 50 % against K. pneumoniae ATCC 700603 and a clinical isolate of E. coli; while compared to Bacillus clinical isolate, was more active than the essential oil. For the rest of microorganisms, the reduction percentages obtained at a concentration of 1 000 ppm varied between 17 and 42 % with ethanolic extract, and 8 to 43 % with essential oil. At concentrations of 100 and 500 ppm antibacterial activity of the extracts was lower. The results indicated that the ethanolic extract and essential oil of C. longa rhizomes have active compounds with antibacterial properties that could be used in future research as a therapeutic alternative for the treatment of infections caused by nosocomial pathogens.

Essential Oil and Ethanol Extract of Oregano (Origanum vulgare L.) from Armenian Flora as a Natural Source of Terpenes, Flavonoids and other Phytochemicals with Antiradical, Antioxidant, Metal Chelating, Tyrosinase Inhibitory and Antibacterial Activity

2019 ◽  
Vol 25 (16) ◽  
pp. 1809-1816 ◽  
Author(s):  
Armenuhi Moghrovyan ◽  
Naira Sahakyan ◽  
Anush Babayan ◽  
Naira Chichoyan ◽  
Margarit Petrosyan ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Essential Oil ◽  
Antibacterial Properties ◽  
Ethanol Extract ◽  
Natural Source ◽  
Origanum Vulgare ◽  
Oregano Essential Oil ◽  
E Coli ◽  
Metal Chelating

Background: Origanum vulgare is used in Armenian cuisine as well as in folk medicine. The aim of this investigation was to determine the peculiarities of the chemical composition of Armenian flora’s oregano essential oil (EO) and ethanol extract (EE) and to characterize mainly antioxidant and antibacterial properties in vitro. Methods: EO of plant aerial parts was extracted by the hydro-distillation method, and the composition was analyzed by gas-chromatography (GC). EE was prepared using ethanol as a solvent. Antioxidant, antibacterial and other biological activity of EO and EE was determined using appropriate methods. Results: The terpenes were found to be the greatest part of oregano EO (β-caryophyllene epoxide - 13.3 %; β- caryophyllene -8.2 %; ο-cymene - 5.2 %). The flavonoid content of EE was 3.9±0.7 mg g-1 catechin equivalents. Antiradical activities of EO and EE expressed with half-maximum inhibitory concentrations were 1057 µg mL-1 and 19.97 µg mL-1. The antioxidant index in the case of using extract (1000 µg mL-1) was 77.3±1.5 %. The metal chelating activity of EE was 74.5±0.2 %. The tyrosinase inhibitory activities of EO, EE and arbutin were 26.5±0.3 %, 6.5±0.2 % and 50±0.1 %, respectively. Both EO and EE of oregano were able to suppress the growth of testmicroorganisms including clinical pathogen isolates, but have moderate antimicrobial activity. EO antibacterial activity against ampicillin-resistant Escherichia coli was similar to the activity against non-resistant strain. EE was not active against ampicillin-resistant E. coli in contrast to non-resistant one. Conclusion: The results indicate high antioxidant and moderate antibacterial as well as high antiradical, metal chelating, tyrosinase inhibitory activity of oregano EO and EE, which can be used as a natural source of terpenes, flavonoids and other phytochemicals in medicine, pharmaceutics, cosmetics and food industry. EO has antibacterial activity also against ampicillin-resistant E. coli.

scholarly journals ANTIFUNGAL AND ANTIBACTERIAL ACTIVITIES OF JUICE AND ETHANOLIC EXTRACTS OF GARCINIA MANGOSTANA L. LEAVES

2019 ◽  
pp. 103-106
Author(s):  
DINIATIK ◽  
ELZA SUNDHANI ◽  
MUHAMMAD TAUFIQ NUGROHO ◽  
EUIS NUR HIKMAH ◽  
RETNO WAHYUNINGRUM ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Skin Infection ◽  
Chronic Wounds ◽  
Inhibitory Concentration ◽  
Ethanolic Extract ◽  
Inhibition Zone ◽  
Antibacterial Activities ◽  
Dilution Method ◽  
Garcinia Mangostana ◽  
E Coli

Objective: Garcinia mangostana is a plant that can be used as a traditional medicine to treat various infectious diseases for the treatment of diarrhea, skin infection, and chronic wounds. The activity as antifungal and antibacterial of juice and ethanolic extract from G. mangostana leaves were investigated. Methods: Juice and ethanolic extract were concentrated using a rotary evaporator to get concentrated extract with rendemen 2.571 and 5.647% (w/w). Juice and ethanolic extract dilution method were employed to evaluate the antifungal activity against Saccharomyces cerevisiae. Ethanolic extract dilution method was used to assess the antibacterial activity against Bacillus subtilis and Escherichia coli. Results: The results of this research showed that juice and ethanolic extract were effective against S. cerevisiae, and the minimum inhibitory concentration was 1000, and 500 mg/mL. Antibacterial activity of the G. mangostana leaves ethanolic extract showed that the action was potential with the inhibition zone in B. subtilis and E. coli. Conclusions: The conclusion of this study is that juice and ethanolic extract of G. mangostana leaves have possible antifungal and antibacterial activity.

scholarly journals Chemical composition and antibacterial activities of the essential oils of Plectranthus glandulosus and Cinnumomum zeylrrnicum from Cameroon

2002 ◽  
Vol 70 (1) ◽  
pp. 93-99 ◽  
Author(s):  
Jirovetz Leopold ◽  
Buchbauer Gerhard ◽  
Ngassoum Martin B. ◽  
Essia-Ngang Jean J. ◽  
Tatsadjieu Leopold N. ◽  
...  
Keyword(s):  
Essential Oil ◽  
Essential Oils ◽  
Active Component ◽  
Solid Medium ◽  
Antibacterial Activities ◽  
Dilution Method ◽  
Aromatic Plants ◽  
E Coli ◽  
And Staphylococcus Aureus ◽  
Type Apparatus

Two aromatic plants have been selected for chemical investigation on account of their antibacterial activities, Plectranthus glandulosus (fresh leaves) and Cinnamomum zeylunicum (dried leaves). P. glandulosus is used as a medicinal plant, while Cinnamomum zeylunicum is used as a common spice in many recipes in Cameroon. The essential oils of the plants have been obtained by hydrodistillation using a Clevenger type apparatus, the yields of oils are about 0.3% and 2.0% respectively.The essential oil of P. glandulosus contains mainly β-thujone (about 31%), p-cymen-8-01 (about 25%) and neral (about 10%) and the essential oil of C. zcylcinicum is characterised by a high percentage of eugenol (85%).These essential oils have been tested against three strains of bacteria, namely Pseudornonas fluorescens. Escherichiu coli and Staphylococcus aureus; qualitative tests (diffusion through solid medium) and quantitative tests (dilution method) have been carried out. P. fluorescens shows a higher resistance to the two oils (MIC values not less than 5000 ppm). E. coli is more sensitive to these oils, while against S uureus the essential oil of C. zcylcinicum still shows a high activity (3500 ppm).The antibacterial activities of the essential oils could be attributed to the components eugenol as the active component of C. zcylcinicum and β-thujone and p-cymen-4-01 of the essential oil of P. glandulosus.

Antioxidant and Antibacterial Activities of Artemisia herba-alba Asso Essential Oil from Middle Atlas, Morocco

2018 ◽  
Vol 16 (S1) ◽  
pp. S48-S54
Author(s):  
Y. Ez zoubi ◽  
S. Lairini ◽  
A. Farah ◽  
K. Taghzouti ◽  
A. El Ouali Lalami
Keyword(s):  
Antibacterial Activity ◽  
Essential Oil ◽  
Foodborne Pathogens ◽  
Food Systems ◽  
Antibacterial Activities ◽  
Culture Collection ◽  
Bornyl Acetate ◽  
Bacterial Strains ◽  
Disk Diffusion Assay ◽  
Different Strains

The purpose of this study was to determine the chemical composition and to evaluate the antioxidant and antibacterial effects of the Moroccan Artemisia herba-alba Asso essential oil against foodborne pathogens. The essential oil of Artemisia herba-alba was analyzed by gas chromatography coupled with mass spectroscopy. The antibacterial activity was assessed against three bacterial strains isolated from foodstuff and three bacterial strains referenced by the ATCC (American Type Culture Collection) using the disk diffusion assay and the macrodilution method. The antioxidant activity was evaluated using the DPPH (2, 2-diphenyl-1- picrylhydrazyl) method. The fourteen compounds of the Artemisia herba-alba essential oil were identified; the main components were identified as β-thujone, chrysanthenone, α-terpineol, α-thujone, α-pinene, and bornyl acetate. The results of the antibacterial activity obtained showed a sensitivity of the different strains to Artemisia herba-alba essential oil with an inhibition diameter of 8.50 to 17.00 mm. Concerning the MICs (minimum inhibitory concentrations), the essential oil exhibited much higher antibacterial activity with MIC values of 2.5 μl/ml against Bacillus subtilis ATCC and Lactobacillus sp. The essential oil was found to be active by inhibiting free radicals with an IC50 (concentration of an inhibitor where the response is reduced by half) value of 2.9 μg/ml. These results indicate the possible use of the essential oil on food systems as an effective inhibitor of foodborne pathogens, as a natural antioxidant, and for potential pharmaceutical applications. However, further research is needed in order to determine the toxicity, antibacterial, and antioxidant effects in edible products.

Bioenhancing Effect of Cow Urine Distillate and Pepper Extract on Antibacterial Activity of Azadirachta Indica Leaves

1970 ◽  
Vol 9 (2) ◽  
pp. 3212-3214
Author(s):  
Pramod Dhakal ◽  
Ankit a Achary ◽  
Vedamurthy Joshi
Keyword(s):  
Antibacterial Activity ◽  
Azadirachta Indica ◽  
Pathogenic Bacteria ◽  
Ethanol Extract ◽  
Watery Diarrhea ◽  
Diffusion Method ◽  
E Coli ◽  
Drug Molecules ◽  
Cow Urine

Bioenhancers are drug facilitator which do not show the typical drug activity but in combination to enhance the activity of other molecule in several way including increase the bioavailability of drug across the membrane, potentiating the drug molecules by conformational interaction, acting as receptor for drug molecules and making target cell more receptive to drugs and promote and increase the bioactivity or bioavailability or the uptake of drugs in combination therapy. The objective of the present study was to evaluate the antibacterial and activity of combination in Azadirachta indica extract with cow urine distillate and pepper extract against common pathogenic bacteria, a causative agent of watery diarrhea. It has been found that Indian indigenous cow urine and its distillate also possess bioenhancing ability. Bioenhancing role of cow urine distillate (CUD) and pepper extract was investigated on antibacterial activity of ethanol extract of Azadirachta indica. Antibacterial activity of ethanol extract neem alone and in combination with CUD and pepper extract were determined the ATCC strains against Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and E-coli by cup plate diffusion method. Ethanol extract of neem has showed more effect on P. aeruginosa, E-coli than S. aureus and K. pneumonia with combination of CUD and pepper extract. CUD and pepper did not show any inhibition of test bacteria in low concentration. The antibacterial effect of combination of extract and CUD was higher than the inhibition caused by extract alone and is suggestive of the bioenhancing role of cow urine distillate and pepper. Moreover, inhibition of test bacteria was observed with less concentration of extract on combining with CUD

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