INTRODUCTION:

Spices have been used for many centuries by various cultures to enhance flavor and aroma of our foods. They have unique aroma and flavor which are derived from compounds known as phytochemicals or secondary metabolites.^1,2 Secondary metabolites of wide variety such as tannins, terpenoids, alkaloids, and flavonoids, which are reported to have a lot of pharmacological activities most of them having antibacterial properties.

Food borne pathogens such as diarrheagenic serotypes of Escherichia coli, Salmonella, Klebsiella, Proteus, Pseudomonas and E. faecalis are widely distributed in nature, causing considerable mortality and morbidity in the population¹. Among the various diarrheagenic serotypes of E. coli, enterohemorrhagic E. coli O157:H7 is implicated in large number of food borne outbreaks in many parts of the world including developed nations. Multi-drug resistant strains of Escherichia coli are widely distributed in hospitals and are increasingly being isolated from community nowadays.³Hence there is an urgent need to find out certain new antimicrobial agents. Since the introduction of antibiotics there has been tremendous increase in the resistance of diverse bacterial pathogens the concentration of scientists is turned towards plant derived products have been used for medicinal purposes for centuries.⁴Herbs and spices are generally considered safe and proved to be effective against many disease.

Plant derived products have been used for medicinal purposes for centuries. At present, it is estimated that about 80% of the world population rely on botanical preparations as medicines to meet their health needs.^5,6,7 Herbs and spices are generally considered safe and proved to be effective against certain ailments. They are also extensively used, particularly, in many Asian, African and other countries. In recent years, in view of their beneficial effects, use of spices or herbs has been gradually increasing in developed countries also.⁸

Spice plants and essential oils extracted from them have become important due to their potential antimicrobial and stimulating effects in the animal digestive system. The antimicrobial effectiveness of mustard, clove, cinnamon and their essential oils were reported for the first time around 1880’s. Antimicrobial effectiveness of spices depend on the kind of spice, its composition and concentration, type and concentrations of the target microorganism, substrate composition, and processing and food storage conditions.^10,11Spice plants have been used traditionally as coloring agents, flavoring agents, preservatives, food additives and as well as anti- parasitic, antihelmintic, analgesic, expectorant, sedative, antiseptic and anti-diabetic substances in many parts of the world. In addition, they possess biological activities such as that of antioxidants and hypocholesterolemics.¹²

In the present study we have evaluated the anti bacterial activity of extracts of few commonly used spices Zingiber officinale (ginger), Piper nigrum (pepper), Allium sativum (garlic), Syzgium aromaticum (Clove) and Cinnamomum verum (cinnamon) against E. coli, Klabsiella, Proteus, Pseudomonas and E. faecalis.

MATERIALS AND METHODS:

Microorganisms:

Escherichia coli, Klebsiella, Proteus, Pseudomonas and E. faecalis were the pathogenic microorganisms included in the study. All the cultures were obtained in pure form from department of microbiology, Saveetha dental college, Chennai and maintained in nutrient agar standard at 4 C.

Preparation spices extracts:

The fresh spices were collected from a local market. The spices were cleaned and washed in sterile distal water. In order to get 100g of each washed spice was crushed with mortar and pestle. The extracts were made to pass through a fine mesh cloth and sterilized using a membrane filter. This extract was considered as the 100% concentration of the extract. The concentrations 75%, 50%, 25% and 10% were made diluting the concentrated extract with appropriate volumes of sterile distilled water.

Garlic extract was made in a different way due to the difficulty to filter the crushed material. One hundred grams of the descaled and cleaned garlic were taken and surface sterilized using ethanol. The ethanol was allowed to evaporate in a sterile laminar flow chamber, and the garlic was homogenised aseptically using a sterile mortar and pestle. The extract was aseptically squeezed out using sterile cheesecloth.

Screening of antibacterial activity [Agar well diffusion technique]:

Broth culture of the test organism compared to 0.5 MacFarland’s standard was prepared. Lawn culture of the test organism was made on the Muller Hinton agar (MHA-Hi media M1084) plates using sterile cotton swab and the plates were dried for15 minutes. A well measuring 4mm depth was made on the agar with sterile cork borer. Different herbal tooth pastes were added to the wells. The plates were incubated at 37°C overnight and the zone of inhibition of growth was measured in millimeters. The tests were repeated triplicate to avoid test error.

RESULTS:

Table 1

Extract	E. coli	Klebsiella	Proteus	Pseudomonas	E.faecalis
Clove	32	26	34	21	28
Garlic	37	30	40	32	35
Cinnamon	29	19	30	12	19
Pepper	32	21	37	18	23
Ginger	29	10	24	09	15
Penicillin G	32	29	35	27	30

Figure 1

DISCUSSION:

Among the five spices extracts tested, three (garlic, pepper and clove) showed antibacterial activity against all the enteric pathogens used in this study (E. coli, Klebsiella, Proteus, Pseudomonas and E. faecalis) is shown in table 1.

Garlic (Allium sativum) showed highest antibacterial activity against all the enteric pathogens in all the concentrations 100%,75% and 50% while the other species showed less antibacterial effect on enteric pathogens. Certain spices like ginger and cinnamon showed very minimal antibacterial effect on enteric pathogens. The values of antibacterial effect of all these spices were compared with standard antibacterial drug penicillin G. Garlic showed 37 mm growth of inhibition against E. Coli, and it showed 30 mm growth of inhibition against Klebsiella, 40 mm growth of inhibition against Proteus, it showed 32 mm growth of inhibition against Pseudomonas, it showed 35 growth of inhibition against E. faecalis. Also garlic showed maximum growth of inhibition when compared to standard antibacterial drug penicillin G against all the enteric pathogens used in this study.

Clove (Syzgium aromaticum) showed second highest antibacterial activity against the enteric pathogens except Proteus. It showed 32 mm growth of inhibition against E. coli, 26 mm growth of inhibition against Klebsiella, 34 mm growth of inhibition against Proteus, 21 mm growth of inhibition against Pseudomonas, and 28 mm growth of inhibition against E. faecalis. But clove was not effective as garlic and also it showed less resistance to enteric pathogens when compared to standard antibacterial drug penicillin G.

Pepper(Piper nigrum) was the next spice to have high antibacterial activity after ginger and clove, which showed good resistance to the pathogens. Similar to clove, pepper showed 32 mm growth of inhibition against E. coli, it showed 21 mm growth of inhibition against Klebsiella, 37 mm growth of inhibition against Proteus, 18 mm growth of inhibition against Pseudomonas, 23 mm growth of inhibition against E. faecalis. Pepper also failed to produce equal amount of antibacterial effect of that produced by standard antibacterial drug penicillin G.

Cinnamon (Cinnamomum verum) was the spice that showed second least antibacterial effect on the enteric pathogens. It showed a variety of results against all the different pathogens and was effective when compared with ginger. Cinnamon showed 29 mm growth of inhibition against E. coli, 19 mm growth of inhibition against Klebsiella, 30 mm growth of inhibition against Proteus, 12 mm growth of inhibition against Pseudomonas, 19 mm growth of inhibition against E. faecalis.

Ginger (Zingiber officinale) was the spice that had least antibacterial activity against enteric pathogens. The results of antibacterial activity of ginger was very low when compared to other spices. Ginger showed 29 mm growth of inhibition against E. coli, 10 mm growth of inhibition against Klebsiella, 24 mm growth of inhibition against Proteus, 09 mm growth of inhibition against Pesudomonas, and 15 mm growth of inhibition against E. faecalis.

Above discussed results and data are given in table 1 and they are also given in a statistical form in figure 1.Spices used in this study have antibacterial activity against the enteric pathogens, the antibacterial activity of the spices were in this order – Garlic, clove, pepper, cinnamon and ginger. Hence the usage of these spices regularly has antibacterial effect and helps in preventing from diseases that are caused by enteric pathogens.

CONCLUSION:

Hence all the spices used in this study have antibacterial activity against the enteric pathogens. Especially garlic showed high antibacterial effect on all the enteric pathogens. The growth of inhibition was greater in garlic when compared with standard antibacterial drug penicillin G.

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Received on 12.08.2018 Modified on 10.10.2018

Research J. Science and Tech. 2019; 11(1):69-72.

DOI: 10.5958/2349-2988.2019.00010.X