بحث منشور للدكتور عبدالامير عبدالله الموسوي حول دراسة الفعالية ضد الجرثومية لاثنين وعشرين نباتا بريا عراقيا

Screening for Antibacterial Activity of Twenty Two Iraqi Wild plants

Abdulameer Abdullah Al-Mussawi

                                             Nursing College / Basra University

                                            E.mail: dr_ameer2006@yahoo.com

Abstract

  The crude ethanolic extracts of twenty two wild plants growing in the Iraqi south regions was evaluated for antibacterial activities against five bacterial species: Gram positive ( Staphylococcus aureus ATCC25923) and Gram negative ( Pseudomonas aeruginosa ATCC27853, Escherichia coli ATCC25922)( Klebsiella pneumonia and Proteus volgars) bacteria by agar well diffusion method . The results showed that the Arnebia decumbens  exhibit broad spectrum activity against all bacterial species.Of 22 plants tested, 10 showed encouraging antibacterial results against one or more species of bacteria .On the other hand 11 plants species have no activity.

Keywords: Antibacterial activity, Agar well diffusion assay, Wild plants

Introduction

  Since the beginning of civilization, survival of the human race was dependent on plants, not only as a source of food and oxygen, but also as a source of natural remedies (Muthu et al., 2010).

  According to world health organization (WHO), more than 80% of the world's population relies on traditional medicines for their primary health care needs. The medicinal value of plants lies in some chemical substances that produce a definite physiologic action on the human body. The most important of these bioactive compounds of plants are alkaloids, flavonoids, tannins and phenolic compounds (Chhetri et al., 2008)  .

  The antimicrobial activity of plants had been received attention many years ago as one of the most effective mechanism for the control of  microorganisms . In Iraq , many studies have been attempted to evaluate the antibacterial activity of some plant extracts (AL-Ani et al ., 1996  ; AL-Thahab,1998).

  Al-Rawi and Chakravarty (1988) study considered the first pioneering study in Iraq classified many wild Iraqi plants.

  We chose this plants in our study because its exist in south of Iraq and no any study investigated in biological activity.

MATERIALS AND METHODS

Plant materials

   Fresh plant parts were collected randomly from various locations in Basra in March and April 2006, 2007,2008 and classified by dr. Ali Aboud Shareef and Louy Hussain Ali, Department of Biology, College of Education. University of Basra (Table:1).

Table 1: Plant Species

Microorganisms

   The test organisms used in this study were as followed: Gram positive (Staphylococcus aureus ATCC25923) and Gram negative bacteria (Escherichia coli ATCC25922) and ( Pseudomonas aerugenosa ATCC27853) These reference bacteria were obtained from the Immunological lab, Biology department , Science college, University of Basra, Iraq. And others clinical Gram negative bacteria Klebsiella pneumonia, Proteus volgars obtained from the Bacteriological lab, Biology department , Education college, University of Basra, Iraq ,(Table:2) and identified according to (Holt , et al., 1994)and (Collee , et al.,1996).

Table 2: Bacterial culture

Preparation of plant extract

   The plant parts of each samples (50g) were air-dried and then powdered. The powder was extracted by reflux with ethanol (250ml) for 15 min followed by  evaporation of combined extracts using rotary evaporator under vacuum. The residue of each extract was kept in refrigerator until use (  Harborne , 1984).

Antibacterial Activity

   Antibacterial activity tested against Gram positive bacteria and Gram negative bacteria by the hole agar diffusion method( Cappuccino and Sherman, 1998). The bacteria were grown on Nutrient agar media. Muller-Hinton agar media were poured into the plates to uniform depth of 5 mm and allowed  to solidify. The bacteria suspensions at 1 x 10⁶cfu ml^-1 (0.1 light density on 540 nm wave length)  were streaked over the surface of Mueller-Hinton agar media using a sterile cotton swab to ensure confluent growth of the organism. The holes made by cookporar, 6 mm in diameter. 100 µL aliquots of the sample 33.3% (v/v), which were then aseptically applied to the surface of agar plates at well-spaced intervals. The plates were incubated at 37 ºC for 24 h and then the inhibition zone diameters were measured.

RESULTS AND DESCUSSION

    In the present investigation study , extracts of 22 plants belonging to different families were screened as antibacterial against five bacterial species, eleven  of which showed activity against at least three of the test bacteria. (Table:3)

   The antibacterial assay results showed that the gram-positive bacteria species are more sensitive than the gram-negative which is in agreement with many previous studies  (Cosentino et al., 1999; Karman et al., 2003).

   Gram negative microorganisms are less susceptible to active compounds than Gram positive ones because they posses outer membrane surrounding the cell membrane (Ratledge & Wikinson,1988) which restricts diffusion of hydrophobic compounds through its lopoplysaccharide covering (Vaara, 1992). This explains the resistance of Gram-negative strains to the lytic action of most extracts exhibiting activity. The negative results obtained against Gram negative bacteria were not unexpected since this class of bacteria is usually more resistant than Gram positive bacteria(Turnbull  et al.,1991).

   The active compounds in plant extracts have many mechanisms to inhibit bacterial growth.  It plays many roles in attacking target sits found in bacteria like cell wall, cell membrane, cellular synthetic processes (DNA,RNA) and protein or enzymes.

       The results showed that the Arnebia decumbens  exhibit high activity against all bacterial species.

   The  activity of Arnebia deumbens extract against all bacteria may come from to active compounds like shikonin and this agreement with previous study  (Singh  et al., 2003).

   The extracts of  Cistanche tubulosa , Astragalus spinosus , Fagonia cretica , Erodium pulverulentum  , Cynomorium coccineum, Sacrophylaria deserti, Bassia eriophora , Reseda lutea, Diplotaxix harra and Rumex nervosus  showed moderate  inhibited the growth of one or more bacteria.

Table 3:

 Inhibition zone of of 11 ethanol extracts exhibit activity against bacterial species

Bacteria: S. aureus = Staphylococcus aureus, E. coli = Escherichia coli, P. aeruginosa = Pseudomonas aeruginosa , K. pneumonia= Klebsiella pneumonia,  P. volgars= Proteus volgars,  control= Ethanol

 On the other hand,  11 plant extracts showed no effect (Table :4).

Table 4: ethanol extracts of some Iraqi wild plants with no antibacterial activity

Bacteria: S. aureus = Staphylococcus aureus, E. coli = Escherichia coli, P. aeruginosa = Pseudomonas aeruginosa , K. pneumonia= Klebsiella pneumonia,  P. volgars= Proteus volgars,  control= Ethanol  

In conclusion, the present investigation study confirmed that  many wild Iraqi plants  contains the potential antibacterial components that may be of great use to the development of new drugs, as a therapy against various diseases.

It is recommended that further studies should be carried out on Iraqi wild plants to further purify the actual bioactive compounds that have the antibacterial activity and to ascertain their toxicity level before recommending for consumption.

REFERANCES

Al-Thahab, A.O.L (1998). The antimicrobial activity of some Iraqi plants on some pathogenic bacteria. M.Sc. thesis college of science . Babylon University .Iraq.

 Al-Ani, A.B. , Nadir, M. T. and Al-Khazraji, N.(1996).The antimicrobial activity of volatile oils isolated from some Iraqi plants .J. of Al- Anbar Univ. 1(1): pp82-86.

AL –Rawi, A. and Chakravarty, H.L. (1988). Medicinal plants of Iraq. Ministry of Agriculture – and Irrigation. 2 and. Ed. Baghdad.

Cappuccino, G. and Sherman, N. (1998). Microbiology: A Laboratory Manual, Benjamin Coming Science Publishing, Calfornia, p.254.

Chhetri, H. P., Yogol, N. S., Sherchan, J., Anupa K.C., Mansoor, S., and Thapa, P. (2008) : Phytochemical and Antimicrobial  Evaluations of Some Medical Plants of Nepal. Kathmandu. J. of  Science , Engineering and Technology.VOL. I, No. V, September, pp 49-54.

Collee, J., Fraser, A., Marmion, B. and Simmons , A. (1996) . Mackie & Macartney practical medical microbiology . 14th(edn.) , Churchill Livingston , New York .

Cosentino, S., Tuberoso, C.I.G., Pisano, B., Satta, M., Mascia, V., Arzedi, E. and Palmas, F. (1999). In-vitro antimicrobial activity and chemical composition of Sardinian Thymus essential oils. Letters in Applied Microbiology. 29: pp130-135.

Harborne, J. (1984) . Phyto chemistry methods : a guide of modern teaching use of plant analysis .2th (edn.) , Chapman and Hill , New York , USA  .

Holt, J.,  Kriej, H., Sneath, P., Staley, J. and William, S. (1994) . Bergey's Manual of Determinative Bacteriology. 9th (edn.) , Williams and Wilkins , Baltimore .

Karman, I., Sahin, F., Gulluce, M., Ogutcu, H., Sengul, M. and Adiguzel, A.(2003). Antimicrobial activity of aqueous and methanol extracts ofJuniperus oxycedrus L. J. of Ethnopharmacology. 85: pp 231- 235 .

Muthu, A.K., Sravanthi, P., Kumar, D.S., Smith, A.A. and Manavalan, R. (2010). Evaluation of antibacterial activity of various extracts of whole plant of Borreria hispida (linn). Int. J. Pharm. Sci. Res.1:pp127-130.             

Ratledage, C. and Wilkinson, S.G.(1988). An overview of microbiallipids.  Microbial lipids. 1:pp3-22.

Singh, B., Sharmaa, M.K., Meghwal, P.R., P.M., Sahua, and Singh, S.(2003). Anti-inflammatory activity of shikonin derivatives from Arnebia hispidissima. Phytomedicine. 10(5):pp 375-380.

Turnbull, P.C.B., Kramer, J.M. Bacillus. In: Barlows A, Hausler JrWJ, Herrmann HD, Isenberg H and Shadomy HJ (eds.)( 1991). Manuals of Clinical Microbiology. 5th Ed. American Society for Microbiology, Washington DC.

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دراسة الفعالية ضد الجرثومية لاثنين وعشرين نباتا بريا عراقيا

عبدالامير عبدالله الموسوي

كلية التمريض ، جامعة البصرة

الخلاصة

تم في هذه الدراسة اختبار تأثير المستخلص الايثانولي لاثنين وعشرين نباتا بريا تنمو في جنوب العراق ضد بعض الانواع البكتيرية الموجبة والسالبة لصبغة كرام وباستخدام طريقة انتشار الاكار .

    استخدمت خمسة انواع بكتيرية احدها موجب لصبغة كرام(ATCC25923Staphylococcus aureus) واربعة سالبة Pseudomonas aeruginosaATCC27853, Escherichia coli ATCC25922 , Klebsiella pneumonia and Proteus volgars)

    بينت النتائج ان نبات   Arnebia decumbens      كان له تأثير واسع الطيف ضد جميع الانواع البكتيرية ، ومن ناحية اخرى اظهرت عشرة مستخلصات نباتية تأثيرا متوسطا ضد واحد او اكثر من الانواع البكتيرية . وقد لوحظ أيضا ان احد عشر نباتا لم تظهر أية فعالية ضد بكتيرية.