Pakistan Journal of Pharmacology Vol.27, No.2, July 2010, pp.53-57

COMPARATIVE ANTIBACTERIAL ACTIVITIES OF SEAWEED EXTRACTS FROM KARACHI COAST, PAKISTAN MUHAMMAD AFZAL RIZVI Hamdard Research Institute of Unani Medicine Faculty of Eastern Medicine, Hamdard University, Karachi -74600, Pakistan ABSTRACT Six species of green seaweeds [Ulva intestinalis Linnaeus, Ulva fasciata Delile, Ulva lactuca Linnaeus, Chaetomorpha antennina (Bory de Saint-Vincent) Kützing, Bryopsis pennata Lamouroux, Codium iyengarii Børgesen, Codium shameelii Nizamuddin, Caulerpa scalpelliformis, Caulerpa taxifolia (Vahl) C Agardh], ten species of brown seaweeds [Dictyota dichotoma var intricata (C Agardh) Greville, Dictyota hauckiana Nizamuddin, Padina antillarum (Kützing) Piccone, Spatoglossum variabile Figari et De Notaris, Stoechospermum polypodioides (JV Lamour) J Agardh, Colpomenia sinuosa (Mertens ex Roth) Derbès et Solier, Iyengaria stellata (Børgesen) Børgesen, Jolyna laminarioides Guimarães, Cystoseira indica (Thivy et Doshi) Mairh, Sargassum ilicifolium (Turner) C Agardh] and also ten species of red seaweeds [Dermonema abbottiae Afaq-Husain, Nizamuddin et Shameel, Gelidium usmanghanii Afaq-Husain et Shameel, Gracilaria corticata (J Agardh) J Agardh, Gracilaria foliifera (Forsskål) Børgesen, Asparagopsis taxiformis (Delile) Trevisan, Cystoclonium purpureum (Hudson) Batters, Botryocladia leptopoda (J Agardh) Kylin, Melanothamnus afaqhusainii Shameel and Osmundea pinnatifida (Hudson) Stackhouse] were collected from the coastal areas of Karachi (Pakistan) and extracted in methanol. These extracts were tested for their antibacterial activity. In comparative study, the brown seaweeds were found to be most active and green seaweeds the least active, while species of red seaweeds occupied an intermediate position. Keywords: Antimicrobial, seaweeds, Karachi coast.

INTRODUCTION Many attempts have been made to investigate the antimicrobial activities of seaweed from the coastal areas of Karachi, Pakistan (Usmanghani et al 1984, Usmanghani et al 1986, Sultana et al 1990, Usmani et al 1991, Siddiqui et al 1993, Hashmi et al 1997, Ali et al 2000, Rizvi et al 2001, Rizvi & Shameel, 2003, Rizvi & Shameel, 2004, Rizvi & Shameel, 2005, Shahnaz & Shameel, 2007, Asma et al 2008) and quite interesting results were obtained. The present study is the continuation of these studies.

MATERIALS AND METHODS The healthy and mature specimens of different species of seaweeds were collected

from various coastal areas of Karachi namely Manora, Sandspit, Buleji and Paradise Point. They were washed with running water to remove epiphytes, attached debris and then soaked in MeOH for antibacterial testing. It was performed against a variety of Gram positive and Gram negative bacteria using agar well diffusion technique. The samples (6 mg) of each crude gummy methanol extract of seaweeds were used for antibacterial test. All pathogenic microbes were clinical isolates and provided by the Department of Microbiology, University of Karachi except Staphylococcus aureus which was obtained from Liaquat National Hospital, Karachi. The pure bacterial cultures were inoculated in nutrient broth and incubated at 37º C for 2-8 h till the turbidity developed. The turbidity of Nutrient Broth (NB Oxoid Uni Path Ltd, England) as obtained in the

Comparative Antibacterial Activities of Seaweed

54 test tubes was compared with McFarland turbidity standard. Test samples of 200 µg/100 µL concentration as well as Dimethyl Sulphoxide (DMSO) were added in their respective wells (Carron et al 1987, Atta-urRahman et al 2001a). The zones of inhibition were measured in mm and compared with reference antibacterial drug: Tetracycline (Sigma USA) and Imipenem.

RESULTS AND DISCUSSION Crude methanol extracts of twenty-six species of seaweeds were tested for their antibacterial activity against four Gram positive and seven Gram negative bacteria (Table 1).Colpomenia sinuosa and Iyengaria stellata appeared to be the most active seaweeds, as they displayed activity against all

Table 1 Comparative antibacterial activities of methanol extracts of seaweeds shown as zone of inhibition in mm. Organisms 1 Chlorophycota Ulva intestinalis Ulva fasciata Chaetomorpha antennina Codium iyengarii Codium shameelii Caulerpa taxifolia Phaeophycota Dictyota dichotoma var intricata Dictyota hauckiana Padina antillarum Spatoglossum variabile Stoechospermum polypodioides Colpomenia sinuosa Iyengaria stellata Jolyna laminarioides Cystoseira indica Sargassum ilicifolium Rhodophycota Dermonema abbottiae Gelidium usmanghanii Gracilaria corticata Gracilaria foliifera Asparagopsis taxiformis Cystoclonium purpureum Botryocladia leptopoda Champia compressa Melanothamnus afaqhusainii Osmundea pinnatifida

Gram positive 2 3

4

5

6

Gram negative 7 8 9

10

11

x x x x x x

x x x 13 -

x x x x 12 x

13 -

x x x x x

x x x -

x x -

x x x 12 x

x x x x x x

14 x -

x x

x

-

x

-

10

-

-

x

x

-

x

x x x x

11 x 12

x x x x

11 -

x x x

x x

x

x x x x

x x 10 x

x x

x -

17 14 9 x x

14 15 13 x

x x

17 14 14 -

x -

14 14 16 13

x

13 12 x x -

13 15 x 14 x

12

x -

10 x x x x x x x x

13 x -

x x x x x x x x

14 -

x x x x x 10 x x

14 x -

x 14 -

x x x x x x x x x

x 9 x x x 14 x x

15 -

x x x x x x -

x

x

x

-

x

x

x

x

x

13

-

1 = Bacillus subtilis, 2 = Corynebacterium diphtheriae, 3 = Staphylococcus aureus, 4 = Streptococcus pyogenes, 5 = Echerichia coli, 6 = Klebsiella pneumoniae, 7 = Proteus mirabilis, 8 = Pseudomonas aeruginosa, 9 = Salmonella typhi, 10 = Shigella dysenteriae, 11= Shigella flexneri, (- = not tested, x = no inhibition., standard drugs: tetracycline & Imipenem; activity: 9-12 = non-significant, 12-15 = low, 15-18 = good, >18=significant).

Rizvi the six tested Gram positive and Gram negative bacteria. These two seaweeds showed similar results, indicating their close taxonomic relationships. Codium shameelii, Cystoseira indica and Botryocladia leptopoda were the next active species, as they exhibited activity against four of the six tested bacterial species. Ulva intestinalis, Stoechospermum polypodioides and Osmundea pinnatifida were the least active algal species among entire group, as they showed a positive activity against only one of the nine tested bacterial organisms. Specific differences were displayed by the different investigated species of Codium, Dictyota and Gracilaria. On the average the brown seaweeds were found to be most active and green seaweeds the least active, while species of red seaweeds occupied an intermediate position. Among Chlorophycota, members of the classes Ulvophyceae and Siphonocladophyceae did not show conclusive differences among themselves. Within Phaeophycota, seaweeds belonging to the class Laminariophyceae were most active and those of the Dictyophyceae least active, while members of the class Fucophyceae occupied an intermediate position. In Rhodophycota, seaweeds of the class Ceramiophyceae were more active than those of the class Nemaliophyceae (Shameel, 2008). Streptococcus pyogenes appeared to be the most sensitive bacterial species, as it was affected by all the six tested algal extracts, while Staphylococcus aureus was influenced by only one the twenty-two tested seaweed extracts. The Gram positive bacteria were found to be slightly more sensitive than the Gram negative ones against the methanol extracts of the tested seaweeds. Brown seaweeds possess a variety of bioactive secondary metabolites and other natural products in their thalli (Ahmad et al 1990, Sastry & Rao 1995, Vlachos et al 1997, Atta-ur-Rahman et al 2001b, Ali & Pervez 2003), which are responsible for their strong antibacterial activity. In several studies,

55 Colpomenia sinuosa and Iyengaria stellata were found to exhibit a variety of bioactivities (Ali et al 2000a, Ara et al 2000, Atta-urRahman 2000), this confirms the present observation as they were found to be most active. Species of Codium, Cystoseira and Botryocladia were also found to contain bioactive compounds in their thalli, which have been isolated by several workers from time to time (Bano et al 1986, 1987, Atta-urRahman et al 1991, Aliya et al 1992, 1993, Ahmad et al 1993, Ali et al 2001, 2002). These natural products may be responsible for their strong antibacterial activities. It is very interesting i.e Stoechospermum polypodioides did not exhibit any activity against eight bacterial species and appeared to be least active among 26 seaweeds. Although, it is very rich in bioactive natural products (Shaikh et al 1990) and its extract has shown a variety of activities (Febles, 1995, Ara et al 2000, Anonymous 2000, Rizvi & Shameel, 2005, Asma et al 2008).

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