ISSN: 0974-2115 www.jchps.com Journal of Chemical and Pharmaceutical Sciences
Evaluation of anti-bacterial activity of the aquatic henna leaves extract in
Hilla City, an in vitro study, Iraq
Nada Khazal Kadhim Hindi1*, Iman Fadhil Abdul-Husin2, Mays Hadi Jebur2,
Zainab Khudhur Ahmad Al-Mahdi3, Ammar Khazaal Kadhim3
1Medical Microbiology, Babylon University, Iraq
2College of Biotechnology, Al-Qasim Green University, Babylon Province, Iraq
3Basic and Medical science Department, College of Nursing, Babylon University, Babylon Province, Iraq *Corresponding author: E-Mail: nadakhazal@yahoo.com, Mobil: 009647601349064 ABSTRACT
Henna leaves have been amassed and selected for antimicrobial towards some human pathogens. The present investigation has been undertaken to find out the effectiveness of aqueous extracts of Lawsonia inermis against some human pathogenic microorganism as well as compare it with wide spectrum antibiotic ciprofloxacin.
In the present study; henna leaves had been extracted with aqueous extraction method. The isolated bacteria were diagnosed based totally on cultural, morphological and biochemical characteristics, hence, the remotes bacterial isolates were as S.aureus, S. epidermidis, S. saprophyticus, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus mutans, Streptococcus fecalis, Streptococcus agalagtia, P.aerugenosa, P. fluroscences, Salmonella typhi, Salmonella typhimurum, Proteus merabilis, Proteus vuligaris, Klebsiella pneumonia, Enterobacter spp, Acinetobacter, Escherichia coli, Serratia spp. The antibacterial activity of the Henna leaves aqueous extract was compare it with wide spectrum antibiotic ciprofloxacin.
The result shows high antibacterial activity of the Lawsonia inermis aqueous extract against tested bacteria with higher a significant different than obtain by ciprofloxacin antibiotics.
Conclusions: Lawsonia inermis exhibit marked antimicrobial activity against bacteria. Based on the results it can be concluded that they can inhibit bacterial growth more than ciprofloxacin antibiotics.
KEY WORDS: anti-bacterial activityو aquatic henna leaves extract.
1. INTRODUCTION
The handiest species in the genus Lawsonia in the own family Lythraceae “Lawsonia inermis” is involved herbal medicine that used for a while. This flowering plant, 2-6m in top. Produces a burgundy dye molecule, lawsone which has an affinity for bonding with protein so, it extensively talking used to dye pores and skin, hair, fingernails, leather-based, silk and wool. Lawsone, is in the main concentrated in the leaves. Leaves of henna plant are whole, opposite, sub-sessile, oval-usual and smooth. Leaves have period of two–3 cm with 1–2 cm width. Henna shrub is especially branched and has greyish-brown barks. The principle makes use of henna are as a cooling agent, astringent, anti-fungal and anti-bacterial herb for the pores and skin and hair. It has also been used as a dye and preservative for hair, pores and pores and skin and fingernails in addition to leather-based completely and clothes. Its center chemical additives are 2-hydroxynapthoquinone (lawsone), mannite, tannic acid, mucilage and gallic acid. Out of these components, the principle one is 2-hydroxynapthoquinone (lawsone). About zero.five-1.5% of henna is fabricated from lawsone. Its bioactive feature is idea to be because of its excessive protein binding capability. In traditional medication, henna plant is used to address many illnesses like oedema, bronchitis, menstrual disease, rheumatism, hemorrhoids or perhaps in jaundice, leprosy, ache, spleen boom, dysentery and pores and pores and pores and skin issues (eight). Henna moreover can be used as an astringent and antihemorragic agent and is also stated for its hypotensive, cardio inhibitory and sedative outcomes. Further, henna is said to expose a few awesome residences which incorporates hypoglycemic (nine), immunostimulant, hepatoprotective (eleven), anti- inflammatory, tuberculostatic, anti-maximum cancers and antioxidant houses.
Aim: The present investigation has been undertaken to find out the effectiveness of aqueous extracts of Lawsonia inermis against some human pathogenic microorganism such as S.aureus, S. epidermidis, S. saprophyticus, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus mutans, Streptococcus fecalis, Streptococcus agalagtia, P.aerugenosa, P. fluroscences, Salmonella typhi, Salmonella typhimurum, Proteus merabilis, Proteus vuligaris, Klebsiella pneumoniae, Enterobacter spp, Acinetobacter, Escherichia coli, Serratia spp as well as compare it with wide spectrum antibiotic ciprofloxacin.
2. MATERIALS AND METHODS
Plant collection:
Preparation of aquatic extracts: The leaves of Lawsonia inermis were obtained from the nearby market of Hilla town – Iraq (March, 2016). The samples turned into cautiously washed beneath strolling faucet water observed by using sterile distilled water after which air dried for two days, pounded. The usage of a mixer grinder and stored in hermetic bottles. Education of aquatic extracts (Aqueous extract) become soaked 50 gram of Lawsonia inermis leaves via 100 ml distillated water, and allowed to face for 72 hr, and sterilized by filtration (the usage of Millipore 0.forty
January - March 2017 162 JCPS Volume 10 Issue 1
ISSN: 0974-2115 www.jchps.com Journal of Chemical and Pharmaceutical Sciences five filter out paper). This extract turned into taken into consideration because the 50% attention of the extract. Oil of cardamom have been accumulated from a retail food keep. The extracts stored in sterile bottles and saved in freezer
at 4oC till further use for screening of antimicrobial activity.
Bacterial Isolates: Different fifteen medical microbial isolates (eight Gram positive, 11Gram negative) had been isolated and recognized by means of the use of traditional biochemical test and Api system (Biomeraux, France) and cultivated in natural culture, at microbiological laboratory/university of Biotecnology/ Al-Qasim green university. In vitro Antibacterial activity testing the use of Agar nicely diffusion assay: Loop full growths from bacterial isolates had been inoculated into nutrient brothincubated at 37°C for 18 hours. The bacterial suspensions have been diluted with everyday saline. regulate the turbidity and examine with widespread tube (McFarland number 0.5) to yield a uniform suspension containing 1.5×108CFU/ml. Cotton swab became dipped and streak into adjustment suspension the entire Mueller-Hinton agar (for all examined bacteria) floor of plates and the plates have been left for one 5 -15 minutes at room temperature to dry. Media have been reduce into four wells (5mm diameter) by using cork borer and add 20? of the cardamom extracts solutions and oil of cardamom (The plates had been performed in triplicates). All plate of the examined organisms was then allowed to incubate at 37°C for in a single day. After 24 h of incubation, every extract turned into cited for region of inhibition for all isolates. The diameters of the region of inhibitions had been measured by means of measuring scale in millimeter (mm).
Antibacterial activity assay: The antibacterial activity became determined through agar disc diffusion. Agar plates have been inoculated with zero.1 ml broth subculture of tested organisms and become spreader with sterile an L- formed rod glass spreader. The antibiotics disks of ciprofloxacin had been upload inside the center of agar plate. (The plates had been finished in triplicates). All plate of the tested organisms changed into then allowed to incubate at 37°C for overnight. After 24 h of incubation, each extract became mentioned for quarter of inhibition for all isolates. The diameters of the region of inhibitions were measured by means of measuring scale in millimeter (mm). Statistical evaluation: Bonferroni test endorsed with the aid of turned into used for statistical evaluation (P ? 0.05) to show if there s any widespread differences among consequences of Lawsonia inermis extract take a look at microorganisms.
Twenty gram positive and gram negative bacteria have been used in this look at to determine the antimicrobial pastime of Lawsonia inermis extract. All bacterial traces have been maintained on freshly prepared blood agar. Then it’s have been identified to the species degree based totally at the stander biochemical and microbiological strategies.
Antimicrobial assay: The antimicrobial activity changed into examined the usage of agar well diffusion approach. Agar plates have been swabbed with 100ml of respected broth way of life (1.5x108CFU/ml), standardized by 0.5 Mac Farland) and have been kept at room temperature for 15min for absorption to take region. Wells of 6mm size have been made with sterile borer in inoculated agar plates and loaded with 100ml of plant extracts. Previous to incubation at 370C for 24hrs. The petri dishes had been saved at room temperature for 15 min. with a purpose to promote diffusion of the extracts into the agar .all the checks were made in triplicate and the suggest diameter of inhibition zones in millimeter.
3. RESULTS AND DISCUSSION
The results were showed in the figure which revealed a higher antibacterial activity of Lawsonia inermis aqueous extract against most common bacterial strains in both type (Gram positive and negative bacteria) with no significant difference (P>0.05) according to activity of the extract between tested bacteria. It is well established that the ?- asarones found in leaf, roots and rhizomes tissues are responsible for almost all of the antimicrobial activities of the Lawsonia inermis. Diacylheptenoid and its Fyrosdiacrosyenin were identified as the constituent responsible for this activity. In this work, aqueous extracts of Lawsonia inermis inhibits the skin pathogens S.aureus, mutans and P.aeruginosa potentially. The antibacterial activity may be attributed to not only a single active principle but to a cocktail of a variety of active principles or alkaloid.
Figure.1. Antibacterial activity of Lawsonia inermis aqueous extract against tested bacteria January - March 2017 163 JCPS Volume 10 Issue 1

ISSN: 0974-2115 www.jchps.com Journal of Chemical and Pharmaceutical Sciences In this search, the result was compare between the result obtained by tested extract and results obtained by ciprofloxacin antibiotic (as most traditional treatment for infection caused by tested bacteria) concerning their
antimicrobial activity against tested bacteria; as reveals in (figure.2).
Figure.2. effect of ciprofloxacin against different types of bacteria
aqueous extracts showed more activity as antimicrobial agent against tested bacterial isolates compare to ciprofloxacin according to inhibition zone diameter and even antibiotic resistant strains ( Pseudomonas aeroginosa, Pseudomonas fluroscences, Klebsiella pneumoniae) and less sensitivity bacteria to ciprofloxacin (Enterobacter spp., Proteus merabilis Proteus vuligaris, Staphylococcus aureus, Staphylococcus epidermidis). These bacterial strain is a mainly multi-drug resistant bacteria that are most commonly found, especially through nosocomial infections.
Considering the findings of this study and comparison with other studies in this field tea extract can be controlled growth of ciprofloxacin resistance bacterial strains which involved in this study within vitro condition.
4. CONCLUSIONS
Lawsonia inermis exhibit marked antimicrobial activity against bacteria. Based on the results it can be concluded that they can inhibit bacterial growth more than ciprofloxacin antibiotics.
REFERENCES
Adcocks C, Collin P, Buttle D.J, Catechins from green tea (Camellia sinensis) inhibit bovine and human cartilage proteoglycan and type II collagen degradation in vitro, J Nutr, 132, 2002, 341–346.
Cos PJ, Arnold V, Dirk VB, Louis M, Anti-infective potential of natural products, How to develop a stronger in vitro proof of concept, J Ethnopharmacol, 106, 2006, 290–302.
Cuong NX, Binh PT, Thuy MTT, Nami NH, Anh HLT, Dati NT, Thao NP, Kiem PV, Huong LM, Isolation of Lawsonia inermis leaves and synthesis of its dimer derivative by cyclic voltammetry, J Chem, 47, 2009, 228–232.
Danial WW, Biostatistics a foundation for analysis in health sciences, 4th ed, John Wilely and Sons Inc, 1988.
Essawi T, and Srour M, Screening of some Palestinian medicinal plants for antibacterial activity J Ethnopharmacol, 70, 2000, 343-349.
Forbes BA, Sahm DF and Weissfeld AS, (Bailey and Scotts Diagnostic microbiology) 12th ed, Elsevier, China, 2007.
Harborne SB, Baxter A. Phytochemical Dictionary, A handbook of bioactive compounds from plants, Tylor and Francis, London, 1995.
Kamal M, Jawaid T, Pharmacological activities of Lawsonia inermis Linn, a review, I J Biomed Res, 2, 2010, 62–68.
Kelmanson JE, Jager AK, van Staden J, Zulu medicinal plants with antibacterial activity, J Ethnopharmacol, 69 (3), 2000, 241-246.
MacGaw L.J, Jaguar A.K and Van Staden J, Isolation of Basarones, an antibacterial and anthermintic compound, from Acores, South African J Bot, 68, 2002, 31-35.
Mikhaeil BR, Badria FA, Maatooq GT, Amer MMA, Antioxidant and immunomodulatory constituents of henna leaves, Z Naturforschung Sec C J Biosci, 59, 2004, 468–476.
NCCLS (National Committee for Clinical Laboratory Standards), Methods for dilution antimicrobial susceptibility tests of bacteria that grow aerobically, Approved Standard M100-S12, Wayne. PA, NCCLS, 2002.
Rahmoun MN, Benabdallah M, Villemin D, Boucherit K, Mostefa-Kara B, Ziani-Cherif C, Choukchou Braham N, Antimicrobial screening of the Algerian Lawsonia inermis (henna), Der Pharma Chemica, 6, 2010, 320–326.
January - March 2017 164 JCPS Volume 10 Issue 1
ISSN: 0974-2115 www.jchps.com Journal of Chemical and Pharmaceutical Sciences Saxena G, McCutcheon AR, Farmer S, Towers GHN, Hancock REW. Antimicrobial constituents of Lawsonia
inermis, J. Ethanol. Pharm, 42, 1994, 95-99.
Sharma VK, Tuberculostatic activity of henna, Lawsonia inermis Linn, Tubercle, 71, 1990, 293–295.
Sidhu MC, Thakur S, Phytochemical and Elemental Exploration of Nothoscordum gracile (Aiton) Stearn for its Medicinal Potential, Journal of Chemical and Pharmaceutical Sciences, 9 (4), 2016, 2627- 2631.
Singh A, Singh DK, Molluscicidal activity of Lawsonia inermis and its binary and tertiary combinations with other plant derived molluscicides, Indian J Exp Biol, 39 (3), 2001, 263-268.
Singh M, Jindal SK, Kavia ZD, Jangid BL, Khem Chand, Traditional Methods of Cultivation and Processing of Henna, Henna, Cultivation, Improvement and Trade, 21-14, Jodhpur, India, Central Arid Zone Research Institute, 2005.
Singh S, Shrivastava NM, Modi NT, Saifi AQ, Anti-inflammatory activity of Lawsonia inermis, Curr Sci, 51, 1982, 470–471.
Srivastava J, Lambert J, Vietmeyer N, Medicinal Plants, an Expanding Role in Development, 1996, 320.
Syamsudin I, Winarno H, The effect of Inai (Lawsonia inermis) leaves extract on blood sugar level, an experimental study, Res J Pharmacol, 2, 2008, 20–23.
January - March 2017 165 JCPS Volume 10 Issue 1