JFS M: Food Microbiology and Safety

Screening for Alternative Antibiotics: An Investigation into the Antimicrobial Activities of Medicinal Food Plants of Mauritius
M.F. MAHOMOODALLY, A. GURIB-FAKIM, AND A.H. SUBRATTY
ABSTRACT: The present study was designed to evaluate the antimicrobial activities of 2 endemic medicinal plants; Faujasiopsis flexuosa (Asteraceae) (FF) and Pittosporum senacia (Pittosporaceae) (PS) and 2 exotic medicinal plants, Momordica charantia (Cucurbitaceae) (MC) and Ocimum tenuiflorum (Lamiaceae) (OT) that forms part of local pharmacopoeia of Mauritius and correlate any observed activity with its phytochemical profile. Aqueous and organic fractions of the leaves, fruits, and seeds of these plants were subjected to antimicrobial testing by the disc diffusion method against 8 clinical isolates of bacteria and 2 strains of fungus. It was found that MC, OT, and FF possessed antimicrobial properties against the test organisms. The MIC for MC ranged from 0.5 to 9 mg/mL and that of FF from 2 to 10 mg/mL and the lowest MIC value (0.5 mg/mL) was recorded for the unripe fruits of MC against E. coli. On the other hand, higher concentration of the unripe MC fruit extract of 9 mg/mL was needed to be effective against a resistant strain of Staphylococcus aureus (MRSA). The antimicrobial effect against MRSA was lost upon ripening of the fruits. The methanolic extract of both MC and FF showed highest MIC values compared to the corresponding aqueous extract, which indicates the low efficacy and the need of higher doses of the plant extract. Phytochemical screening of the plants showed the presence of at least tannins, phenols, flavonoids, and alkaloids, which are known antimicrobial phyto-compounds. In conclusion, the observed antimicrobial properties would tend to further validate the medicinal properties of these commonly used endemic medicinal and food plants of Mauritius. Keywords: antimicrobial, endemic, exotic, medicinal plants

erbal medicine is holistic medicine (Bagchi 2001). It is, in fact, able to look beyond the symptoms to the underlying systemic imbalance; when skillfully applied by the trained practitioner, herbal medicine offers very real and permanent solutions to very real problems, many of them seemingly intractable to pharmaceutical intervention (Gogtay and others 2002). The use of and search for drugs and dietary supplements derived from food plants have accelerated in recent years. Ethnopharmacologists, botanists, microbiologists, and natural-products chemists are combing the Earth for phytochemicals and leads that could be developed for treatment of infectious diseases (Balandrin and others 1985). Traditional healers have long used plants to prevent or cure infectious conditions; Western medicine is trying to duplicate their successes. For instance, medicinal herbs now constitute the most rapidly growing segment of the total U.S. pharmaceutical market. Figures show that about 25% of all prescription sold in the United States are from natural products, while another 25% are from structural modifications of a natural product (Mathews and others 1999). Indeed knowledge gained from the use of medicinal herbs and their active ingredients serve as the foundation for much of modern pharmacology (Ang-Lee and others 2001). Additionally, the development of modern chemistry permitted the isolation of chemicals from medicinal herbs have served as drugs or starting materials for the synthesis of many important drugs used today. Drugs such as aspirin, digitalis, morphine, and quinine were all originally isolated
MS 20090654 Submitted 7/7/2009, Accepted 1/19/2010. Authors are with Dept. of Health Sciences, Faculty of Science, Univ. of Mauritius, Reduit, Mauritius. Direct inquiries to author Mahomoodally (E-mail: f.mahomoodally@uom.ac.mu).

Introduction

or synthesized from materials derived from plants (Mathews and others 1999). Mauritius also has a long-standing tradition in use of phytomedicines. Furthermore, being a tropical island, Mauritius has a very rich and diverse flora and many of the plant species are used for their medicinal virtues. Nonetheless, even with this vast array of data, few medicinal plant species of Mauritius have been scientifically evaluated for their possible medicinal application. Among the vast number of medicinal plants in current use in Mauritius, are Momordica charantia, an exotic vegetable (Subratty and others 2005) and 3 commonly used endemic/indigenous medicinal plants; Faujasiopsis flexuosa (Lam.) C. Jeffrey; Pittosporum senacia (Pittosporaceae); and Ocimum tenuiflorum (Lamiaceae) (Gurib-Fakim and others 1996a, 1996b, 1996c) were evaluated in the present study for their antimicrobial properties. Therefore, as part of a systematic study on the flora of Mauritius, these 4 plants were subjected to standard antimicrobial screening in an attempt to assess their current use in the traditional medicine of Mauritius. In addition, this investigation also endeavors to compare the antimicrobial activities of different extracts by finding their minimum inhibitory concentration.

Materials and Methods

Preparation of extracts
Unripe fruits, leaves and seeds of Momordica charantia (Cucurbitaceae) (MC) used in the study were obtained from a commercial source throughout the island. Leaves of the endemic plants, Pittosporum senacia (Pittosporaceae) (PS), and Faujasiopsis flexuosa (Asteraceae) (FF) were collected from Conservation
Vol. 75, Nr. 3, 2010JOURNAL OF FOOD SCIENCE M173

C 2010 Institute of Food Technologists R doi: 10.1111/j.1750-3841.2010.01555.x

Further reproduction without permission is prohibited

M: Food Microbiology & Safety

Screening for alternative antibiotics . . .

Management Areas (Maccabe forest) situated in the upper humid region of Petrin and Forest-Side conservation area, Mauritius. Exotic plant Ocimum tenuiflorum (Lamiaceae) (OT) were obtained from the Univ. of Mauritius farm. The Curator of the national Herbarium, at the Mauritius Sugar industry research institute (MSIRI) confirmed the identity of the plants. Voucher specimens were deposited at the Herbarium collection of the Dept. of Chemistry, Faculty of Science, Univ. of Mauritius. organisms using sterile Pasteur pipette. The plates were allowed to dry. The disks containing the plant extract were transferred using flamed but cooled forceps onto the surface of the seeded agar plates. They were sufficiently spaced to prevent the resulting zones of clearing from overlapping. The extractive solvent (water) was used as negative control. The plates with the organisms were incubated for 24 h. Standard antibiotic was used as positive control for comparison. Zone of inhibitions was measured to the nearest millimeter.

Extraction
Crude aqueous extract of the plant material was used as per local pharmacopoeia. Freshly collected plant materials were either airdried or dried in a drying cabinet at 50o C for 5 to 7 d. Total of 10 g of the dried plant materials (fruit, leaves, and seeds) of the plant species were separately crushed and ground into fine powders using a food blender. The outer rind of MC fruit was removed before drying. The solvent was distilled off under reduced pressure to afford crude plant extract. The paste was collected in water for examination. Percentage yield was calculated and the paste-like water suspension was diluted for further experiments.

Determination of minimum inhibitory concentration (MIC) values

The MIC of each plant extract was determined by a slight modification of the tube dilution method (Omoregbe and others 1996). In a set of 10 sterile capped test tubes using the extraction solvent as diluent, serial dilutions were made from the different extracts to get graded concentration in milligrams per milliliter and a tube containing only diluent as the sensitivity control. Sterile filter paper disks (6 mm in diameter) were impregnated with the different dilutions of the plant extract were aseptically transferred to the surface of the inoculated plates using flamed but cooled forceps. The disks were sufficiently spaced to avoid overlapping of zones of inhibition. Test culture The test organisms used for the screening of antimicrobial activ- The MIC of the different plant extracts that inhibited the growth of ity were Bacillus cereus, Bacillus subtilis, E. coli, Staphylococcus au- the test organism other than inhibition due to the diluent (water) reus, methillin resistant Staphylococcus aureus (MRSA), Enteroccous was taken as MIC. faecalis, Pseudomonas aeruginosa, and Salmonella typhimurium. Phytochemical screening Stock cultures of the bacteria and fungus used were obtained from Fruits, seeds, leaves, and twigs where appropriate were subjected Bacteriology Section of the central laboratory Candos, Victoria Hosto a thorough phytochemicals screening using standard protocols pital. Cultures were maintained as nutrient agar slants in screw (Mahomoodally and others 2005) to detect the presence of the folcapped bottles and stored at 4 C. All cultures were checked for vilowing secondary metabolites: alkaloids, coumarins, terpenes, anability and purity by regular plating. Test cultures were prepared thraquinones, tannins, phenols, leucoanthocyanins, flavones, and by transferring a loop full of bacteria from stock culture nutrient saponins. broth and incubated at 37 C for 24 h. Fungus were transferred into freshly prepared dextrose agar plates and incubated at 25 C for Results 3 d. The fungi used in the study were Aspergillus niger and Candida esults obtained for the antimicrobial tests performed on aquealbicans. ous and methanolic extracts of OT, MC, PS, and FF are presented in Table 1. Results showed that the crude aqueous and Antimicrobial bioassay procedure methanolic extracts of the plants showed broad spectrum of acSterilized filter paper disks (6 mm in diameter) were placed in tivity, being active almost to both the Gram-positive and Gramappropriate concentration of each plant extracts in screw-capped negative organisms in the antimicrobial assays. MC leaves, twigs, seeds, and unripe fruit showed variable degrees bottles. The disks (made from Whatman nr 1) were allowed to absorb the plant extracts. Plates of MuellerHinton sensitivity agar of antibacterial and antifungal activities against the test microor(oxoid) were aseptically inoculated with broth cultures for the test ganisms. Our results also showed that the aqueous unripe fruit of

M: Food Microbiology & Safety

Table 1 --- Antimicrobial activities of aqueous and methanolic extracts of PS, OT, FF, and MC against the test microorganisms. PS Stda 14 12 10 13 7 9 10 L 5(4) 7(2) 13(6) 6(4) () () () () () () T 3(2) () 6(8) () () () () () () () L 7(5) 8(4) (4) 7(8) (6) 10) () 6() () () OT T () () () (6) (10) () () () () () Diameter of zone of inhibition (mm)b FF L 5(5) 10(9) 8(8) 6(5) () 5(3) 8(6) 10(8) () () T 3(3) 4(2) 5(3) 5(5) () 3() 6(3) 7(5) () () L () 3(2) 11(9) () () 4(2) 3(3) 10(8) () () UF 4(3) 8(6) 15(10) 17(16) 5(2) 4(2) 7(7) 13(12) 3(2) 8(6) MC RF 2() 6(3) 10(5) 12(8) () 5(2) 5(2) 10(8) 2() 7(4) T () 3(2) 10(8) 9(4) () 5(3) 3() 5(3) () () S () 2() 10(8) 9(9) () () 3() 5() () ()

Microorganism BC BS EC SA MRSA EF PA ST AN CA

(Inhibition zone of methanolic extract.) a Standard antibiotic Ampicillin tested at a concentration of 10 g/disc (oxoid). b Zone of inhibition; average duplicates of 3 independent experiments. = Complete lack of activity. L = leaf extract; T = twigs extract; UF = unripe fruit; RF = ripe fruit; BC = Bacillus cereus; BC = Bacillus subtilis; EC = E. coli ; SA = Staphylococcus aureus; MRSA = methicillin resistant Staphylococcus aureus; EF = Enterococcus faecalis; PA = Pseudomonas aeruginosa; ST = Salmonella typhimurium; AN = Aspergillus niger ; CA = Candida albicans.

M174

JOURNAL OF FOOD SCIENCEVol. 75, Nr. 3, 2010

Screening for alternative antibiotics . . .

MC possess the most potent antimicrobial properties (100% active), being active almost to both Gram-positive and Gram-negative bacteria, and the 2 fungal strains. The inhibition zone ranged from 3 to 17 mm and 2 to 16 mm for the aqueous and methanolic fruit extract, respectively. However, twigs of MC gave the poorest inhibition zones, being active to 50% of the test organisms. The inhibition zones produced by the unripe fruits of MC against S. aureus, S. typhimurium, and E. coli were greater than the standard antibiotic ampicillin. On the other hand, the methanolic extracts of aerial parts of MC resulted in lower antimicrobial activity compared to the aqueous extracts. Only fruit of MC was found to possess antifungal activities against A. niger and C. albicans with inhibition zones of 3 to 8 mm. The antimicrobial activity of MC fruit was partly lost or significantly reduced upon ripening of MC fruit that resulted to smaller range of inhibition zones (2 to 12 mm) compared to the unripe fruit extract (2 to 17 mm). Both leaves and twigs of FF showed inhibitory effects on the test organisms. However, no antifungal activities against A. niger and C. albicans were observed. The inhibitory zone (10 mm) of the crude aqueous extract of the leaves of FF is comparable to the 10 g/disc standard pure antibiotic ampicillin for S. typhimurium. The leaves extract of FF was found to be the most active extract with greater bactericidal effects (inhibition zones 3 to 10 mm) against the test organisms compared to the twigs (inhibition zones 3 to 7 mm). Unlike MC extracts, FF did not yield any antimicrobial effect against MRSA. However, similar decrease in the antimicrobial activities was observed for the methanolic extract compared to the corresponding aqueous extract of FF. Leaves and twigs of PS and OT showed moderate activity. The inhibitory zones ranged from 2 to 13 mm and 2 to 8 mm for PS and OT, respectively. Of particular interest is the zone of inhibition of PS against E. coli (3 mm for the aqueous extract and 6 mm for the methanol extracts). It is to be noted that the aqueous leaf extracts of PS showed higher inhibitory activities as compared to the standard antibiotic ampicillin (10 g/mL discs). The crude methanol extract OT leaf and twigs was also found to possess activity against MRSA (6 and 10 mm zones of inhibition, respectively). of OT, respectively; 1 to 8 mg/mL and 4 to 8 mg/mL for leaves and twigs of FF, respectively; and 0.5 to 8 mg/mL, 3 to 8 mg/mL, 1 to 8 mg/mL, and 1.5 to 8 mg/mL for unripe fruits, ripe fruits, twigs, and seeds of MC, respectively. For the MRSA strains, high concentrations of the leaves extracts of the plants were needed to inhibit growth of the bacteria. It is to be noted that the MIC for MC ranged from 0.5 to 9 mg/mL and that of FF from 2 to 10 mg/mL and the lowest MIC value (0.5 mg/mL) was recorded for the unripe fruits of MC against E. coli. On the other hand, a higher concentration of the unripe MC fruit extract of 9 mg/mL was needed to be effective against the MRSA strains. The antimicrobial effect against MRSA was lost upon ripening of the fruits. The methanolic extract of both plant parts showed highest MIC values compared to the corresponding aqueous extract, which indicates the low efficacy and the need of higher doses of the plant extract.

Phytochemical analysis of the medicinal plants

Table 3 shows the results from phytochemical screening of the 6 medicinal plant extracts. Tannins were present in all the tested extracts whereas anthraquinones were absent in all tested extracts. From these data, it can be deduced that the phytochemical profile of the leaves for almost all the plants does not correlate with that of the twigs. However, only MC, the phytochemicals in the both plant parts corresponds to each other.

Discussion

MIC
All extracts that showed antibacterial activities by the disc diffusion method were further tested by the broth dilution assay to determine the MIC. Results of the MICs of the different plant extracts are shown in Table 2. The MIC value for aqueous extract of leaves and twigs of PS ranged from 0.5 to 16 mg/mL and 2 to 12 mg/mL, respectively; 4 to 8 mg/mL and 1 to 8 mg/mL for leaves and twigs

nterest in higher plant extracts exhibiting antimicrobial activity has increased in recent years, and several reports on this subject have been published (Farnsworth 1994; Benzi and Ceci 1997; Bagghi 2001; Rangasamy and others 2007; Thakurta and others 2007). In the present study, the 4 medicinal plants were selected as they have been documented to possess antimicrobial properties and used traditionally by the local population to treat minor infections (Gurib-Fakim and others 1996a, 1996b, 1996c). To this effect, standard antimicrobial assays were carried against key pathogenic bacteria and fungi to validate their use in the traditional medicines and also the consumption MC as a common vegetable in Mauritius. The exotic plants; MC and OT and the endemic medicinal plants; PS and FF possess antimicrobial properties against the test microorganisms. In the local traditional pharmacopoeia of Mauritius, MC is extensively used to treat DM. However, it is also used against diarrhea, stomach colic, and to combat intestinal infections. On the other hand, FF is known to possess astringent properties and is used by the local population against dysentery (Narod

Table 2 --- Minimum inhibitory concentrations (mg/mL) PS, OT, FF, and MC plant parts for the 10 strains of microorganisms. Minimum inhibitory concentration (mg/mL) PS Microorganism BC BS EC SA MRSA EF PA ST AN CA L 10 (8) 8 (12) 2 (4) 6 (8) () () () () () () T 12 (8) () 6 (6) () () () () () () () L 4 (6) 8 (8) (8) 8 (8) (8) (6) () 8 () () () OT T () () () (8) (8) () () () () () L 1 (1) 4 (6) 4 (4) 6 (8) () 2 (4) 8 (8) 4 (6) () () FF T 8(8) 8 (8) 6 (7) 4 (6) () 4 (6) 6 (7) 4 (6) 6 () () L () 4 (8) 1 (4) () () 7 (4) 4 (6) 2 (8) () () UF 5 (8) 3 (8) 0.5 (1) 2 (4) 9 (8) 5 (8) 2 (8) 1 (4) 2 (4) 3 (6) MC RF 8 () 6 (8) 4 (8) 4 (8) () 8 (8) 6 (3) 2 (4) 4 () 8 (8) T () 6 (8) 1 (2) 4 (8) () 8 (4) 8 () 2 (4) () () S () 8 () 1.5 (3) 5 (6) () () 8 () 3 () () ()

(MIC of methanolic extract.) L = leaf extract; T = twigs extract; UF = unripe fruit; RF = ripe fruit; BC = Bacillus cereus; BC = Bacillus subtilis; EC = E. coli ; SA = Staphylococcus aureus; MRSA = methicillin resistant Staphylococcus aureus; EF = Enterococcus faecalis; PA = Pseudomonas aeruginosa; ST = Salmonella typhimurium; AN = Aspergillus niger ; CA = Candida albicans. = Complete lack of activity.

Vol. 75, Nr. 3, 2010JOURNAL OF FOOD SCIENCE

M175

M: Food Microbiology & Safety

Screening for alternative antibiotics . . .

Table 3 --- Phytochemical components of FF, OT, and PS leaves and twigs; and MC fruits. Plants Part used Alkaloids Flavonoid Tannins Leucoanthocyanins Anthraquinones Terpenes Phenols Coumarins Saponins FF PS MC OT Leaf Twigs Leaf Twigs Fruits Leaf Leaf Twigs + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +

+ = presence; = absence.

and others 2004), which is classified as an infectious disease (Bloom 1994). Additionally, it has been found that species from the same genus of OT possess antimicrobial properties (Aqil and others 2005; Chah and others 2006). Recently, Aqil and others (2005) reported the inhibitory activities of Ocimum species against clinical isolates of -lactamase producing MRSA and methicillin-sensitive S. aureus (MSSA). Therefore, these documented properties are in conformity with the results obtained in this study, since these plants have been shown to possess the ability to inhibit growth of both pathogenic Gram-positive and Gram-negative bacteria. However, the plant parts differ significantly in their activity against the test microorganisms. The differences observed in the antimicrobial activities assays suggest the susceptibility of the test microorganisms to various secondary metabolites present in these medicinal plants. The composition of these secondary metabolites in turn varies from species to species, climatic conditions, and the physiological state of developments of the endemic plants (Hussain and Deeni 1991). Available reports tend to show that alkaloids and flavonoids are the responsible compounds for the antimicrobial activities in higher plants (Cordell and others 2001). Moreover, it is also claimed that secondary metabolites such as tannins and other compounds of phenolic nature are classified as active antimicrobial compounds (Mitscher and others 1972; Rojas and others 1992). In this study, it has been found that these 4 plants possess important phytochemicals such as saponins, flavonoids, tannins, and alkaloids Additionally, Welininda and others (1982) have isolated steroidal glycosides, alkaloids, momordicosides, and insulino-mimic proteins from MC. Recently, Braca and others (2008) identified 25 components from MC, representing 90.9% of the oil. The main constituents were trans-nerolidol, apiole, cis-dihydrocarveol, and germacrene D. Furthermore, they tested the oil for its antibacterial and antifungal activities and found that Staphylococcus aureus was the most sensitive microorganism (Braca and others 2008). Therefore, the presence of these phytochemicals could to some extent justify the observed antimicrobial activities in the current study. It is worthwhile to note that since the current antimicrobial evaluation was done using the same aqueous preparation as prescribed by the traditional healers, these results would support the way people use these herbal remedies. MC fruit was found to have bactericidal effect on 1 strain of Staphylococcus aureus (MRSA), which is resistant to the antibiotic methicillin. In Mauritius nosocromial infection caused by MRSA is a serious problem in many hospitals of the island. These MRSA are difficult to treat because they are also multiresistant and up to now there are no satisfactory antimicrobial drugs (Bartzokas 1984; Beck and others 1986; Berman and others 1986). Therefore, with respect to the present result, MC fruits seem to be a potential tool to combat the problem of MRSA. Moreover, in the light of the emergence of resistant strains of pathogens to known drugs and the high cost of health care in developing countries, the WHO recommendation

suggests the use of medicinal plants as alternative antibiotics in these countries (Mathews and others 1999). Thus MC fruit, which is widely used as a vegetable in the diet should not be ignored or neglected as dietary adjuncts. Findings from the present study also showed that MC extracts gave the lowest MIC values. In addition, MC plant parts seem to have a broader spectrum of antimicrobial activities against the test microbes. These results clearly prove the possible greater efficacy of MC extract at a lower dose than the endemic plant FF. Results from present study also showed that methanolic extract of both plants gave poor inhibition zones. Indeed, Cragg and others (1994) reported that there are great differences between the activities of aqueous and organic extracts in their anti-HIV screening: about 34% of the plants were active in aqueous extract and only 4% in organic extract. Moreover, the antibacterial substance within these plants in the current study seemed to be most prominent in the leaves and least in the twigs, and the best inhibitory activity was observed for Gram-positive bacteria. This may be attributed to the fact that cell wall in Gram-positive bacteria consists of a single layer, whereas Gram-negative cell wall is a multilayered structure bounded by an outer cell membrane (Yao and Moellering 1995). On the other hand, the extracts were not highly efficient against the test fungi. Such results indicate that antibacterial agents are more common in the plants studied than antifungal agents. This may be attributed to structural differences between prokaryotic bacteria and eukaryotic fungal agents; antimicrobial agents should bind to sterols in eukaryotic membrane so as to exhibit their action, whereas this step is not needed for bacterial cells (Medoff and Kayashi 1993; Ali-Stayeh and others 1998). Results from the current study also show that methanol extracts of these plants tend to give poor inhibition zones.

M: Food Microbiology & Safety

Conclusion

t is possible that these endemic/exotic medicinal plant extracts can be used as antibacterial agents in food or other ingredients. However, the exact mechanism of antibacterial effects needs to be further examined for potential uses. In conclusion, the current study advocates the need for continuing screening for antimicrobial agents from local endemic plants of Mauritius.

Acknowledgment
The authors are grateful to the Tertiary Education Commission and The Univ. of Mauritius for Financial support. We are also thankful to Dr. Issack, Senior specialist/Clinical Microbiologist, Victoria Hospital, Ministry of Health & Quality of Life for providing us with clinical isolates.

References
Ali-Stayeh MS, Yaghmour RMR, Faidi YR, Salem K, Al-Nuri MA. 1998. Antimicrobial activity of 20 plants used in folkloric medicine in Palestinian area. J Ethnopharmacol 60:26571.

M176

JOURNAL OF FOOD SCIENCEVol. 75, Nr. 3, 2010

Screening for alternative antibiotics . . .

Ang-Lee MK, Moss J, Yuan CS. 2001. Herbal medicines and perioperative care. J Am Med Assoc 286:20816. Aqil F, Khan MS, Owais M, Ahmad I. 2005. Effect of certain bioactive plant extracts on clinical isolates of beta-lactamase producing methicillin resistant Staphylococcus aureus. J Basic Microbiol 45:10614. Bagchi AK. 2001. Alternative medicine-old wine in a new bottle. J Ind Med Assoc 98:3323. Bagghi AK. 2001. Alternative medicine-old wine in a new bottle. J Ind Med Assoc 98:3323. Balandrin MF, Klocke JA, Wurtele ES, Bollinger WH. 1985. Natural plant chemicals: sources of industrial and medicinal materials. Sci 228:115460. Bartzokas CA. 1984. Control and eradication of methicillin resistance Staphylococcus aureus on surgical unit. New Eng J Med 311:1422. Beck WD, Berger-Bachi B, Kayser FH. 1986. Additional DNA in methicillin-resistance Staphylococcus aureus and molecular cloning of mec-specific DNA. J Bacter 165:3738. Benzi G, Ceci A. 1997. Herbal medicines in European regulation. Phamacol Res 35:35562. Berman DS, Schaefer S, Aimberkoff MS. 1986. Tourniquets and nosocomial methicillin resistant Staphylococcus aureus infection. New Eng J Med 311:5145. Bloom SR. 1994. Tooheys Medicine: a textbook for students in health care professions. London, U.K.: Longman Group Ltd. p 1550. Braca A, Siciliano T, DArrigo M, Germano MP. 2008. Chemical composition and antimicrobial activity of Momordica charantia seed essential oil. Fitoterapia 79:1235. Chah KF, Eze CA, Emuelosi CE, Esimone CO. 2006. Antibacterial and wound healing properties of methanolic extracts of some Nigerian medicinal plants. J Ethnopharmacol 104:1647. Cordell GA, Qunia-Beattie M, Farnsworth MR. 2001. The potential of alkaloids in drug discovery. Phytother Res 15:183205. Cragg GM, Boyd MR, Cardellina I, Newman DJ, Snader KM, McCloud TG. 1994. Ethnobotany and drug discovery: experience of the U.S. National Cancer Institute. Chichester, U.K.: Chadwick & Marsh Press. Farnsworth NR. 1994. Ethnopharmacology and drug development. Ciba Foundation Symposium 185:4259. Gogtay NJ, Bhatt HA, Dalvi SS, Kshirsagar NA. 2002. The use and safety of nonallopathics: Indian medicines. Drug Safe 25:100519. Gurib-Fakim A, Gueho J, Sewraj MD. 1996a. Plantes medicinales de Maurice. Tome 1. Ed. Locean Indien, Stanley, Mauritius: Rose Hill. p 1498. Gurib-Fakim A, Gueho J, Sewraj MD, Dulloo E. 1996b. Medical ethnobotany of some weeds of Mauritius and Rodrigues. J Ethnopharmacol 39:17585. Gurib-Fakim A, Gueho J, Sewraj MD, Dulloo E. 1996c. Medicinal plants of Rodrigues. Int J Phamacolo 34:214. Hussain HSN, Deeni Y. 1991. Plants in Kano ethnomedicine: screening for antimicrobial activity and alkaloids. Int J Pharmacol 29:516. Mahomoodally MF, Gurib-Fakim A, Subratty AH. 2005. Antimicrobial and phytochemical profiles of endemic medicinal plants of Mauritius. Pharm Biol 43:237 42. Mathews HB, Lucier GW, Fisher KD. 1999. Medicinal herbs in the United States: research needs. Environ Health Perspect 107:7738. Medoff G. Kayashi GS. 1993. Mode of action of antifungal drugs. New York. p 32549. Mitscher LA, Leu RP, Bathala MS, Wu WN, Beal JL. 1972. Antimicrobial agents from higher plants. Introduction, rationale and methodology. Lloydia 35:15766. Narod F, Gurib-Fakim A, Subratty AH. 2004. Biological investigation into Antidesma madagascarieniss lan.(Euphorbiaceae), Faujasiopsis flexosa (lan.), C.Jeffy (Asteraceae), Tod dalia asiatica(L.) lam and Vepris lanceoloata (lam) G.Don (Rutaceae). J Cell Mol Biol 3:1521. Omoregbe RE, Ikuebe OM, Jhimire IG. 1996. Antimicrobial activity of some medicinal plants extracts on Escherichia coli, Salmonella paratyphi and Shigella dysenteriae. Afri J Med Sci 25:3735. Rangasamy O, Raoelison G, Rakotoniriana FE, Cheuk K, Urverg-ratsimamanga S, Quetin-Leclercq J, Gurib-Fakim A, Subratty AH. 2007. Screening for anti-infective properties of several medicinal plants of the Mauritians flora. J Ethnopharmacol 109:3317. Rojas A, Hernandez L, Pereda-Miranda R, Mata R. 1992. Screening for antimicrobial activity of crude drug extracts and pure natural products from Mexican medicinal plants. J Ethnopharmacol 35:27583. Subratty AH, Mahomoodally MF, Gurib-Fakim A. 2005. Bitter melon: an exotic plant with medicinal values. Food Nutr J 35:1437. Thakurta P, Bhowmik P, Mukherjee S, Hajra Patra A, Bag K. 2007. Antibacterial, antisecretory and antihemorrhagic activity of Azadirachta indica used to treat cholera and diarrhea in India. J Ethnopharmacol 22:60712. Welininda J, Avvidoon G, Gylfe E, Hellings B, Karlsson E. 1982. The insulin-releasing activity of the tropical plant Momordica charantia. Acta Biol Med 41:122940. Yao J, Moellering R. 1995. Antibacterial agents: manual of clinical microbiology. Washington, D.C. p 128190.

Vol. 75, Nr. 3, 2010JOURNAL OF FOOD SCIENCE

M177

M: Food Microbiology & Safety