WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Rudrangshu et al.

World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 2.786

Volume 4, Issue 4, 1058-1073.

Research Article

ISSN 2278 – 4357

COMPARATIVE STUDY OF ANTIMICROBIAL ACTIVITY OF ALOE VERA GEL AND ANTIBIOTICS AGAINST ISOLATES FROM FAST FOOD Rudrangshu Chatterjee*, Dushyant Singh, Amita Gaurav Dimri, Ankita Pandita, Shiwani Chaudhary and M.L.Aggarwal Department of Microbiology, Shriram Institute for Industrial Research 19, University Road, Delhi-110007.

Article Received on 18 Jan 2014, Revised on 13 Fab 2015, Accepted on 10 Mar 2015 Nov 2014

ABSTRACT Aloe vera is reputed to have medicinal properties. For centuries, it has been used for an array of ailments such as mild fever, wounds and burns, gastrointestinal disorders, diabetes, sexual vitality and fertility problems to cancer, immune modulation, AIDS and various skin

*Correspondence for

diseases. In present study antibacterial activity of Aloe vera gel

Author

extracts were tested against common foodborne pathogens i.e. Bacillus

Rudrangshu Chatterjee

cereus, Staphylococcus aureus, Escherichia coli and Pseudomonas

Department of Microbiology, Shriram

aeruginosa. For this purpose commonly available twelve different fast

Institute for Industrial

food samples i.e Chow mine, Momo, Spring roll, Chat, Pani Puri,

Research 19, University

Aalu-Tikki, Veg Sandwich, Burger, Paovaji, Veg Patties, Bhelpuri and

road, Delhi-110007.

Bread Pakora were collected from suburb of Delhi. Antibiotic resistance and susceptibility pattern of above isolates were also studied

against five clinically significant antibiotics (Meropenem, Erythromycin, Cefoperazone, Ciprofloxacin and Doxycycline). Meropenem was found to be most effective antibiotic followed by Cefoperazone with 100% efficacy. In case of Erythromycin 100% efficacy was shown against gram positive bacteria while gram negative bacteria have shown resistance against it. Aloe vera leaves gel were extracted with four different solvents like Methanol, Ethanol, Chloroform and Di- chloro methane (DCM), however Gram-positive isolates were found 100% susceptible with methanol extract whereas no inhibition was observed with chloroform and DCM extract. Moderate sensitivity observed with ethanol extract against both Gram -positive as well as Gram -negative bacterial isolates. Aloe vera gel showed greater

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antibacterial activity against Gram-positive bacterial isolates as compared to Gram-negative bacterial isolates. KEYWORDS: Aloe vera, Fast food, Antibiotics, Susceptibility, Resistance, Agar well diffusion assay, MAR index. INTRODUCTION Food is a chemically complex matrix, and predicting whether, or how fast, microorganisms will grow in any given food is difficult. Most foods contain sufficient nutrients to support microbial growth. Several factors encourage, prevent, or limit the growth of microorganisms in foods. The busy and hectic life schedule has opened the way for the fast food industry in most parts of the world. The traditional or conventional way of cooking is over and the fast food joints are visible everywhere. Fast food does not only include the traditional fast food items like pizza, burger or French fries but it also includes Chinese as well as Indian. Microbial food safety is an increasing public health concern worldwide. It is estimated that each year in the United States there are approximately 76 million food borne illnesses,[22] cases are caused by Campylobacter spp., nontyphoidal Salmonella, pathogenic Escherichia coli all colonize the gastrointestinal tracts of a wide range of wild and domestic animals, especially animals raised for human consumption.[23] Food contamination with these pathogens can occur at multiple steps along the food chain, including production, processing, distribution, retail marketing and handling or preparation. It was reported that numerous epidemiological reports have implicated foods of animal origin as the major vehicles associated with illnesses caused by food-borne pathogens.[29, 33] Microorganisms in fast and traditional fast foods are responsible for many human diseases. Fast foods sold in a restaurant or store with low quality preparation and served to the customer in a packaged form for takeout/take away. In most fast food operations, menu items are generally made from processed ingredients prepared at a central supply facility and then shipped to individual outlets where they are reheated, cooked (usually by microwave or deep frying) or assembled in a short amount of time, fast food are often very high in calories, saturated fat and sodium that can make us fatter, clog our arteries and send our blood pressure soaring. It was suggested that ensure that food should cook thoroughly to the correct temperature because proper cooking kills almost all dangerous microorganisms. The Contamination of the food supply with pathogens and its persistence, growth, multiplication or toxin production has emerged as an important public health concern. Over two hundred different diseases are

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known to be transmitted by the food. Food-related bacteria constitute a heterogeneous group with their original habitats extending to all ecological niches where food for human consumption is produced and handled. The contaminated surfaces play a crucial role in relation to potential transmission of pathogens to food in food processing, catering and domestic environment. Several studies indicates that various bacteria, including Escherichia coli, Staphylococcus aureus B.cereus and Salmonella spp., survive on hands, sponges/cloths, utensils and currency for hours and days after initial contact with the microorganisms. In some other studies, the extent of bacterial survival and cross contamination between hands and food or various kitchen surfaces have been quantified. This routine practice of food borne pathogens have been a significant driving force in accelerating the emergence of Antibiotic Resistant bacteria that subsequently are transferred from contaminated surfaces and animals to humans through food chain. In fact, antimicrobial resistance has increased dramatically during the last decade among foodborne pathogens. In India, infectious diseases accounts for high proportion of health problems. Morbidity and mortality due to these infections continues to be a major problem, especially amongst children. Infections due to a variety of bacterial etiologic agents, such as pathogenic Escherichia coli, Staphylococcus aureus, Salmonella sp. Bacillus cereus, Enterobacter sp. are most common.[21] In the present time multiple drug resistance in microbial pathogens become a serious health problem to humankind worldwide.[27] Synthetic drugs are not only expensive and inadequate for the treatment of diseases but also often associated with adulterations and side effects. Therefore, there is need to search new infection fighting strategies to control microbial infections. However, the past record of rapid, widespread emergence of resistance to newly introduced antimicrobial agents indicates that even new families of antimicrobial agents will have a short life expectancy while there are some advantages of using medicinal plants. Plants have an important source of medicine for thousands of years. Even today, world health organization estimates that up to 80% of people remedies such as herbs for their medicines. Aloe vera is an ornamental and medicinal plant. The name of Aloe vera was derived from the Arabic ‘ALLOEH’ meaning ‘BITTER’ because of bitter liquid found in the leaves. It is also known as ‘LILY OF DESERT’. Aloe vera is a stem less or very short –stemmed succulent plant growing to 60-100 cm tall, spreading by offsets. The leaves are thick and fleshy, green to grey-green, with some varieties showing white flecks on the upper and lower stem surfaces. It is being used therapeutically, since Roman times and perhaps long before.[6, 20] Different properties being ascribed to the inner colorless leaf gel and to the exudates from the

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outer layers. Aloe has a history of traditional use by Native Americans for stomach disorders and intestinal disorders including constipation, hemorrhoids, and colitis and colon problems. It is said to be a natural cleaner, powerful in penetrating tissues, relieving pain associated with joints and muscles, bactericidal, a strong antibiotic, virucidal when in direct contact with long periods, fungicidal, anti-inflammatory, instrumental in increasing circulation to the area, breaking and digesting dead tissue and moisturizing tissues. The skin absorbs Aloe vera up to four times faster than water, it appears to help pores of the skin open and receive moisture and nutrients of the plants. Additionally, numerous constituents within Aloe vera have demonstrated enhancement of immune system functioning within the body. Aloe also has the ability to stimulate macrophages.[7] The term ALOE refers to a solid residue obtained by evaporating the latex derived from the outer layers of the plant leaf. Taxonomists now refer to Aloe barbadensis as Aloe vera. Among more than 360 Aloe species, A. vera (A. barbadensis miller) has been the most popular in both folk and officinal medicine. More than 75 ingredients have been identified from the gel,[10,

12]

each of which may have a range of

mechanism of actions, acting synergistically or individually to explain more than 200 different constituents notably mucopolysaccharides, enzymes, sterols, prostaglandins, fatty acids, amino acids and a wide variety of vitamins and minerals. It contains several potentially active bioactive compounds including salicylates, magnesium lactate, acemannan, lupeol, campestrol, β-sitosterol, aloin A and anthraquinones.[9] In addition Aloe vera contains at least seven super-oxide dismutases with antioxidant activity. The efficacy of Aloe liquid as an antibacterial agent is shown to have a wide range against Gram positive and Gram negative bacteria. The antimicrobial agents of Aloe vera gel was reported to effectively kill or greatly reduce or eliminate the growth of Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus pyogenes, Pseudomonas aeruginosa, Escherichia coli, Propionibacterium acne, Helicobacter pylori and Salmonella typhi.[19,

28, 32, 34]

Whole leaf components are

proposed to have direct antibacterial properties include anthraquinones and saponins; [32, 34] while polysaccharides have been attributed within direct activity through the stimulation of phagocytic leucocytes to destroy bacteria.[19, 28] The aim of the present study was to evaluate the effects of an A. vera gel and leaf extract as well as to check the resistance and susceptibility patterns of three clinically significant antibiotics against different bacterial isolates from fast food samples collected from suburb of Delhi. The study involves; (a) Collection of fast food samples from different location (b) Isolation of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus

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cereus, from fast food samples (c) Extraction of gel from Aloe vera leaves with four different solvents like Methanol, Ethanol, Chloroform and Di- chloro methane (DCM) (d) Determination of susceptibility and resistance pattern of bacterial isolates against Aloe vera gel extracts as well as five different antibiotics by agar well diffusion assay (e) Interpretation of the results for the benefit of human welfare by increasing general awareness among the people. MATERIALS AND METHODS Collection of fast food samples: Wide mouth PET jars sterilized by gamma-radiation were used for sampling of seven different fast food samples are shown in Table 1. The lid of the jar was removed by maintaining all aseptic conditions. The samples were kept in an ice pack to prevent any changes in the microbial flora of the samples. The samples of food were transported to the lab for testing in vertical position maintaining the temperature 1-4°C with ice pack enveloped conditions were reached to the laboratory starting the analysis within 6 hrs of collection. Isolation and identification of Pathogens Detection of E. coli: For the detection of E.coli 25 g homogenized sample was diluted with 225 ml of Nutrient broth and then incubated at 37 0C for 24 hrs. Subcultured on the MacConkey agar and Eosin Methylene blue agar plates. Plates were observed for characteristic colonies such as pink colonies on MacConkey agar and green metallic sheen colonies on Eosin Methylene blue agar plates and further confirmation was done by Gram’s staining and biochemical test using HiIMViC test kit (Hi media).[14] Detection of Pseudomonas aeruginosa: For the detection of Pseudomonas aeruginosa 25 g homogenized sample was diluted with 225 ml of cetrimide broth and then incubated at 37 0C for 48 hrs. Subcultured on the plates of cetrimide agar Plates were observed for characteristic green colonies and further confirmation was done by Gram’s staining biochemical test.[16] Detection of Bacillus cereus: For the detection of Bacillus cereus 25 g homogenized sample was diluted with 225 ml of Buffer Peptone Water and then incubated at 370C for 24hrs. Subcultured on the Mannitol Yolk Polymixin agar plates. Plates were observed for characteristic colonies such as pink colonies with zone and further confirmation was done by Gram’s staining and biochemical test.[15]

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Detection of Staphylococcus aureus: For the detection of Staphylococcus aureus 25 g homogenized sample was diluted with 225 ml of cooked meat medium (with 10% salt) and then incubated at 370C for 24hrs. Subcultured on the Mannitol salt agar and Baird parker agar plates. Plates were observed for characteristic colonies such as yellow colonies on Mannitol salt agar plates and black colonies on Baird parker agar plates. Further confirmation was done by Gram’s staining and biochemical test.[17] Extraction of gel from Aloe vera leaves: The fully expanded leaves of Aloe vera were selected from the plants, washed with distilled

water and were subjected to surface

sterilization with 70% Iso propyl alcohol. The Parenchymatous covering of the leaves were peeled and the gel drained out. Slurry was formed with the help of pestle and mortar, leaf gel was dried in the oven at 80°C for 48 h. and then powdered. 10 grams of this powder was soaked in 100ml. of solvents namely Methanol, Ethanol, Chloroform and Di- chloro methane (DCM) for 24 h. The contents were then filtered through Whatman filter paper no. 1 and the filtrate was evaporated to dryness. This dried extract was further powdered and then dissolved in distilled water to make the working solution having 10mg/ml concentration. Solvent controls (Methanol, Ethanol, Di-chloro methane & Chloroform) were prepared in the similar manner. Antibiotics and their solutions: Five antibiotics like – ERYTHROMYCIN (Erythromycin tablet IP (250mg), Mfd by: Rajasthan Drugs & Pharmaceuticals ltd.), MEROPENEM (Meropenem Injection IP (1g), Mfd by: Elder Pharmaceuticals Ltd.), CEFOPERAZONE (Cefoperazone Injection (1 g), Mfd by: Park Pharmaceuticals), DOXYCYCLINE (Doxycycline

hyolate

IP

(100mg),

Mfd

by:

Jagsonpal

Pharmaceutical

limited),

CIPROFLOXACIN (Ciprofloxacin tablet-IP 500mg, Mfd by: Cipla Limited) were used to check susceptibility and resistance pattern of above four bacterial isolates. All these antibiotics were obtained from local pharmacy store and working solution having 10mg/ml concentration of each antibiotic was used for the study. Inoculum Preparation: Above four bacterial isolates were sub cultured on non selective nutrient agar slants. The bacterial cultures were incubated overnight at 37°C. 0.5 McFarland density of bacterial isolates was adjusted using normal saline (0.85% NaCl) using densitometer to get bacterial population of 1.0 x 108 cfu/ml.

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Agar Well Diffusion Assay (Zone of Inhibition Evaluation): Antimicrobial activity of isolated strains was evaluated by agar well diffusion assay. 100µl of each of the adjusted cultures were mixed into separate 100 ml of sterile Muller Hinton Agar (MHA), mixed well and poured into sterile petri plates. These were allowed to solidify and then individual plates were marked for each individual culture. Each plate was punched to make wells of 6 mm diameter with the help of sterile cork borer at different sites of the plates. 100 µl of each supernatant as well as 100 µl of respective antibiotic solutions were pipette into the well in assay plates. Plates were incubated overnight at 37oC. Following incubation, petri-plates were observed for the inhibition zones, diameters of which were measured by using Vernier Calipers. RESULTS & DISCUSSION Fast foods are ready-to-eat foods sold by vendors especially in streets and other similar places. These fast foods provide an affordable source of nutrients to the population who appreciate the food due to its taste, low price and availability at the right time. Food-borne illnesses associated with the consumption of street vended fast foods, restaurant, homemade kitchen food has been reported in several places in India and elsewhere.

[9]

Present study

dealt with the microbiological analysis of commonly available twelve fast food samples collected from different locations of suburb of Delhi. These samples were analyzed for the presence of E.coli, Pseudomonas aeruginosa, Bacillus cereus and Staphylococcus aureus. The above four isolated bacterial strains (Table 1) were biochemically characterized and then evaluated for their susceptibility and resistance patterns against Aloe vera gel extracts. Antimicrobial activity of A. vera gel was also compared with five standard antibiotics used in the study. Table 1: Microbiological profiling of Fast Food samples collected from different location Sample Code S-1 S-2 S-3 S-4 S-5 S-6 S-7 S-8

Name of the sample Chow mine Momo Spring roll Chat Pani puri Aalu-Tikki Burger Veg

Location Dwarka Janakpuri C1 Janakpuri Kamala nagar Palam Tilak nagar Azadpur Karol Bag

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Escherichia coli/25 g (A) Present Present Present Present Present Present Absent Present

Bacillus cereus/25 g (B) Present Present Present Absent Absent Present Present Present

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Pseudomonas aeruginosa/25 g (C) Absent Absent Absent Present Present Absent Absent Absent

Staphylococcus aureus/25 g (D) Present Absent Absent Present Present Absent Present Present

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S-9 S-10

Sandwich Paovaji Veg Patties

S-11

Bhelpuri

S-12

Bread Pakora

World Journal of Pharmacy and Pharmaceutical Sciences

Noida Moolchand Chandni chawk Malkaganj

Absent Present

Absent Present

Absent Absent

Present Present

Present

Absent

Present

Absent

Present

Present

Absent

Present

It was observed that E.coli, Bacillus cereus and Staphylococcus aureus were prominently present however Pseudomonas aeruginosa was found to be present in Chat, Pani puri, and Bhelpuri. The antibacterial property of Aloe vera gel extracted using different solvents showed varying degree of response towards the selected pathogens (Table 2 and Table 3). Table 2: Zone of inhibition against Aloe vera gel extract Zone of inhibition* (In mm) against Aloe vera gel extract Sample marked as Methanol Ethanol Chloroform DCM S-1(A) 0 10 9 0 S-1(B) 16 16 0 0 S-1(D) 20 18 0 0 S-2(A) 0 14 9 0 S-2(B) 19 15 0 0 S-3(A) 0 10 0 0 S-3(B) 19 14 0 0 S-4(A) 0 10 8 0 S-4(C) 0 12 0 0 S-4(D) 21 17 0 0 S-5(A) 0 10 8 0 S-5(C) 0 12 0 0 S-5(D) 22 18 0 0 S-6(A) 0 10 0 0 S-6(B) 17 20 0 0 E.coli positive control 0 15 9 0 Pseudomonas positive 0 14 0 0 control S.aureus positive control 24 22 0 0 B.cereus positive control 21 20 0 0 Solvent Control 0 0 0 0 *Zone of inhibition in mm. Diameter including well diameter of 6.0 mm S: sample number (A) E.coli, (B) B.cereus, (C) P. aeruginosa, (D) S.aureus Table 3: Zone of inhibition against Aloe vera gel extract Sample marked as S-7(B) S-7(D) S-8(A)

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Zone of inhibition* (In mm) against Aloe vera gel extract Methanol Ethanol Chloroform DCM 15 15 0 0 20 15 0 0 0 14 7 0

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S-8(B) 18 19 0 S-8(D) 19 18 0 S-9(D) 20 22 0 S-10(A) 0 10 9 S-10(B) 20 0 0 S-10(D) 17 16 0 S-11(A) 0 11 9 S-11(C) 0 12 0 S-12(A) 0 10 8 S-12(B) 20 20 0 S-12(D) 16 22 0 E.coli positive 0 15 9 control Pseudomonas 0 14 0 positive control S.aureus positive 24 22 0 control B.cereus positive 21 20 0 control Solvent Control 0 0 0 *Zone of inhibition in mm. Diameter including well diameter of 6.0 mm

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

S: sample number (A) E.coli, (B) B.cereus, (C) P. aeruginosa, (D) S.aureus Using ethanol extracts the zones of inhibition ranged from 15 to 22 mm being maximum for S.aureus and minimum for E.coli (10 mm). Methanol extract exhibited maximum antibacterial activity against S.aureus (24 mm) followed by B.cereus (21mm). With respect to individual pathogens, Ethanol extract showed susceptibility against gram positive as well as gram negative isolates while, significantly least inhibition was observed for E.coli (7 to 9 mm) with Chloroform extract. In case of solvent control no zone of inhibition was observed. Hence the antimicrobial activity was totally of the Aloe vera extract obtained. All four bacterial isolates showed resistance against DCM extract as no zone of inhibition was observed. Fig 1 demonstrated the percentage of zone of inhibition of bacterial isolates against Aloe vera gel extract in different solvents used in the study. Generally methanol extract showed greater antibacterial activity against gram positive isolates. These differences may be attributed to the fact that the cell wall in gram positive bacteria consists of a single layer, whereas the gram negative cell wall is a multi-layered structure and quite complex.[24]

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Fig 1: Zone of inhibition against Aloe vera gel extract This result could be responsible for the popular use of A. vera gel to relieve many types of gastrointestinal irritations,[8,

11]

the gel is also said to promote wound healing due to the

presence of some components like anthraquinones and hormones,[7] which posses antibacterial antifungal and antiviral activities. However, most of the constituents are found in the gel and not in the leaf, hence the gel is likely to be more active than the leaf. The gel possesses 100% inhibitory effect on gram positive bacteria. Other workers have reported antibacterial properties of ethanol extracts of A. vera gel against the pathogens selected in the study. [1, 32] There was no antimicrobial activity was reported using aqueous extract of A. vera leaves. [25] Nearly all of the identified components from plants active against micro-organisms are aromatic or saturated organic compounds and are most often obtained through initial ethanol or methanol extraction.[4] This explains higher antimicrobial activity of ethanol and methanol extracts observed in the study. A lower antimicrobial action against Gram-negative bacteria as compared to Gram-positive organism could be explained to be due to the presence of additional lipopolysaccharide layer in the former. Table 4 showed the comparative study of A. vera gel extract with standard antibiotics (Figure 2). Table no 4: Comparative study of A. vera gel extract with standard antibiotics Antibiotic Erythromycin Cefoperazone Doxycycline Ciprofloxacin A. vera gel extract Methanol A. vera gel extract Ethanol A. vera gel extract Chloroform

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Gram positive (%) 100 100 0 28 100 94 0

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Gram negative (%) 0 100 0 73 0 100 60

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Fig 2: Comparative study of A. vera gel extract with standard antibiotics against Gram positive and Gram negative bacteria Table 5: Zone of inhibition against antibiotics Sample marked as S-1(A) S-1(B) S-1(D) S-2(A) S-2(B) S-3(A) S-3(B) S-4(A) S-4(C) S-4(D) S-5(A) S-5(C) S-5(D) S-6(A) S-6(B) S-7(B) S-7(D) S-8(A) S-8(B) S-8(D) S-9(D) S-10(A) S-10(B)

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Zone of inhibition* (In mm) against Antibiotics Meropenem Erythromycin Cefoperazone Ciprofloxacin Doxycycline 28 0 28 24 0 22 18 11 0 0 32 10 28 0 0 23 0 21 18 0 24 9 26 0 0 14 0 10 22 0 28 11 21 0 0 29 0 18 0 0 33 0 17 27 0 25 10 29 20 0 31 0 23 21 0 31 0 23 0 0 28 12 12 0 0 20 0 10 10 0 22 14 14 16 0 25 11 12 0 0 27 12 15 0 0 20 0 18 12 0 25 10 10 18 0 22 12 12 0 0 24 15 19 0 0 30 0 15 14 0 27 9 20 0 0

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S-10(D) 25 8 15 S-11(A) 22 0 15 S-11(C) 30 0 12 S-12(A) 25 0 10 S-12(B) 20 13 10 S-12(D) 24 10 14 E.coli positive 31 0 18 control Pseudomonas 28 0 20 positive control S.aureus positive 28 14 20 control B.cereus positive 25 14 22 control *Zone of inhibition in mm. Diameter including well diameter of 6.0 mm

0 12 10 14 0 0

0 0 0 0 0 0

15

0

18

0

17

0

15

0

Fig 3: Zone of inhibition of bacterial isolates against different antibiotics In present study it was found that the most effective antibiotic for gram positive as well as gram negative is Meropenem followed by Cefoperazone with 100% efficacy (Fig 3). In case of Erythromycin 100% efficacy were shown against gram positive bacteria however gram negative bacteria have shown resistance against it. In our study high level of resistance was reported to Doxycycline as all isolates were found to be resistant against this antibiotic (Table 5). Susceptibility rate was intermediate i.e. 54.55% against Erythromycin as well as Ciprofloxacin. The fast increase in the appearance of the multi-drug resistant bacteria is a major problems faced by medicine and science today. Antimicrobial resistance in bacterial population has increased worldwide and its susceptibility patterns show substantial

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geographic variation as well as differences in population and environment. The Multiple Antibiotic Resistance (MAR) index of profile was performed to evaluate the heath risk of the environments. Multiple antibiotic resistance index (MAR) (number of antibiotics to which test isolate displayed resistance divided by total number of antibiotic to which the test organism has been evaluated for sensitivity) for each test isolate was calculated.

[18]

An

isolate with a value of MAR > 0.2 is an indicator of the area with a high risk of contamination where the antibiotics are frequently used. MARs of different bacterial strains isolated in the current study are represented in Table 6. Table 6: MAR Index of bacterial Isolates E.coli Isolates *MAR Value B.cereus Isolates *MAR Value Pseudomon as Isolates *MAR Value S.aureus Isolates *MAR Value

S-1(A)

S-2(A)

S-3(A)

S-4(A)

S-5(A)

S-6(A)

S-8(A)

S-10(A)

S-11(A)

S-12(A)

0.4

0.4

0.4

0.6

0.4

0.4

0.4

0.4

0.4

0.4

S-1(B)

S-2(B)

S-3(B)

S-6(B)

S-7(B)

S-8(B)

S-10(B)

S-12(B)

0.4

0.4

0.4

0.2

0.4

0.2

0.4

0.4

S-4(C)

S-5(C)

S-11(C)

0.4

0.6

0.4

S-1(D)

S-4(D)

S-5(D)

S-7(D)

S-8(D)

S-9(D)

S-10(D)

S-12(D)

0.4

0.2

0.4

0.4

0.4

0.4

0.4

0.4

In the current study, only two B. cereus isolates and one S. aureus strains were shown to have MAR value is equal to 0.2. Rest bacterial isolates were having the MAR up to 0.6 showing high degree of resistance. This study reveals the applicability of the multiple antibiotic resistance (MAR) with the aim to identify the level of contamination within the commonly available fast foods. This very high level of contamination indicates a potential breakdown of hygiene at various stages of the food processing and distribution chain and/or a lack of refrigeration of food materials to be used for fast food preparation. The presence of multidrug resistant strains is alarming, because such strains lead to a higher fatality rate than sensitive ones. Obviously, the prudent use of antimicrobial agents is a prerequisite for the minimization of the emergence of drug-resistant bacteria, but such prudence in itself is not enough to control this emerging public health threat.

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CONCLUSION An alarming consequence has been occurred due to widespread emergence of resistance among microorganisms against available antibiotics. The present study has revealed the importance of natural products to control antibiotic resistant bacteria, which have been a threat to human health. As global antibiotic resistance by bacteria is becoming an interesting public health concern and the race to discover the new antibacterial agent is on, Aloe vera gel along with its identified compounds with promising antibacterial activity could be used as an alternative herbal remedy. Hence, it can be concluded that the compounds isolated from Aloe vera gel extracts could be recommended for human trials (in proper dosage) against different bacterial pathogens at the same time this study can led to beneficially assist in the identification of alternate drug to control these multi-drug resistant bacterial strains. REFERENCES 1. Agarry, O.O.; Olaleye, M.T.; Bello-Michael,C.O. Comparative Antimicrobial Activities of Aloe vera gel and leaf. Afr. J. Biotechnol, 2005; 4(12): 1413-1414. 2. Asma Bashir1, Bushra Saeed1, Talat .Y. Mujahid2 and Nayar Jehan Comparative study of antimicrobial activities of Aloe vera extracts and antibiotics against isolates from skin infections, 2007. 3. Belo SE, Lorena RG, Patricia MB. Moisturizing effect of cosmetic formulations containing Aloe vera extract in different concentrations assessed by skin bioengineering techniques, Skin. Res. Technol., 2006; 12: 241–246. 4. Cowan, M.M. Plant Products as Antimicrobial Agents. Clin. Microbiol. Rev, 1999; 12(4): 564-582. 5. C. L. Little, S. Walsh, L. Hucklesby, S. Surman-Lee, K. Pathak, Y. Gatty, M. Greenwood, E. De Pinna, E. J. Threlfall, A. Maund and C. H. Chan, J Food Prot, 2007; 70: 2259-2265. 6. Crosswhite, F.S.; Crosswhite, C.D. Aloe vera plant symbolism at the threshing floor. Desert plants, 1984; 6: 43-50. 7. Davis, H.R. Aloe vera: A scientific approach. Published by Vantage Press, New York, 1997; 35. 8. Foster S. Aloe vera: The succulent with skin soothing cell protecting properties. Herbs for Health magazine, 1999. 9. Ghosh, S.; Playford, R.J. Bioactive natural compounds for the Treatment of Gastrointestinal disorders. Clin. Sci. (Great Britain), 2003; 104: 547-556.

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