In Vivo and In-Vitro Evaluation of Antimicrobial Activity of Peel Extracts of Red Dragon Fruit (Hylocereus polyrhizus)

 

Yogita Temak*1, Pravin Cholke1, Akshay Mule2, Akahay Shingade2, Sudam Narote2, Aditee Kagde2, Rutuja Lagad2, Vaishnavi Sake2

1HOD, Department of Pharmaceutical Analysis and Pharmaceutical Chemistry,

Loknete Shri Dadapatil Pharate College of Pharmacy, Mandavgan Pharata, Shirur, Pune. (M.S.)

2Students of Third Year B. Pharmacy, Loknete Shri Dadapatil Pharate College of Pharmacy,

Mandavgan Pharata, Shirur, Pune. (M.S.)

*Corresponding Author E-mail: yogitatemak2013@gmail.com

 

ABSTRACT:

This study aimed to characterization and evaluates the effectiveness of red dragon fruit (Hylocereus polyrhizus) peel extracts as an antimicrobial agent. Red dragon fruit peel extracts were obtained by maceration using solvent at pH 5. Phytochemical characteristics, total phenols, antioxidant, and antimicrobial activity of the peel extracts were observed. Antioxidant and antimicrobial activities of the extracts were associated with high phytochemical compounds and total phenols contained in the extracts. Red dragon fruit peel extracts with various percentages prepared and their physicochemical characteristics, nutrients, antioxidant activity, and microbiological profile were analyzed. The current study is to evaluate the susceptible antimicrobial activity of crude 50% hydro alcoholic Peel Extracts of Red Dragon Fruit (Hylocereus polyrhizus) against different clinical isolates (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, A. niger and Candida albican) of microbial disease so as to trip up on the other alternatives and overcome the upcoming era of increasing microbial resistance. Furthermore, the results of the in vivo assay on thirty mice model showed that the treatment of mice with the 100mg/ml of aqueous extract and Chloramphenicol have more effectiveness than other concentrations and the number of bacteria P. aeroginosa was reduced significantly (P ≤ 0.001) during the treatment days. Results showed that the red dragon fruit peel extracts have an Antimicrobial Activity.

 

KEYWORDS: antimicrobial, red dragon fruit peel extract, pitaya etc.

 

INTRODUCTION:

Dragon Fruits:

Pitaya usually refers to fruit of the genus Stenocereus, while pitaya or dragon fruit refers to fruit of the genus Hylocereus, both in the Cactaceae family. Sweet pitayas come in three types, all with leathery, slightly leafy skin.

·       Hylocereus undatus (Pitaya blanca or white-fleshed pitaya) has pink-skinned fruit with white flesh. This is the most commonly seen "dragon fruit".

·       Hylocereus costaricensis (Pitaya roja or red-fleshed pitaya, also known as Hylocereus polyrhizus) has red-skinned fruit with red flesh.

·       Hylocereus megalanthus (Pitaya amarilla or yellow pitaya, also known as Selenicereus megalanthus) has yellow-skinned fruit with white flesh.

 

Early imports from Colombia to Australia were designated Hylocereus ocampensis (supposedly, the red fruit) and Cereus triangularis (supposedly, the yellow fruit). It is not quite certain to which species these taxa refer, though the former is probably the red pitaya.

 

The fruit normally weighs from 150 to 600 grams (5.3 to 21.2 oz); some may reach 1 kilogram (2.2 lb).


 

Hylocereus undatus (Pitaya blanca or white-fleshed pitahaya)

 

Hylocereus costaricensis (Pitaya roja or red-fleshed pitaya,

 

Hylocereus megalanthus (Pitaya amarilla or yellow pitaya,

 

 

 

Picture No.01: Plants of Pitaya

 


EXPERIMENTAL:

Reagent and Materials:

All the reagents in these experiments along with triple distilled water were of analytical grade. All the chemicals were purchased from Merck Research Chemicals pvt. Ltd. Red Dragon Fruit (Hylocereus polyrhizus) were purchased from local market. For in vivo activity required mice supply by Savitribai Phule Pune University, Animal Experiments Department, Pune.

 

Apparatus:

Automatic B.O.D. INCUBATOR, Model No.TC141 and ANTIBIOTIC ZONE READER Model No. FG 456 made by LABGO are also used this research work.

 

Antibacterial activity (In-Vitro):

In radial or 2-D technique petri-dishes of agar are prepared by pouring melted agar media previously inoculated with selected microorganism after the solidification of agar cups are made with the help of borer and cups are filled with solution of suitable conc. of sample and standards respectively and are inoculated at 37°C for 24 hr. The anti-microbial agent diffuses through the agar around its cup and produces a characteristic zone of inhibition of the microorganism sensitive to the sample, the diameter of which can be measured.

 

Table No 01: Antibacterial activity ofcrude 50% hydro alcoholic Peel Extracts of Red Dragon Fruit (Hylocereus polyrhizus)

Extracts

Zone of inhibition at 200μcg/Ml (in mm.)

 

E.coi

B. Subtilis

S. aureus

A. niger

C. albicans

A1

24

25

26

15

22

A2

20

23

25

16

21

A3

20

24

25

19

22

A4

25

26

23

20

21

A5

24

23

26

21

22

A6

20

22

24

18

23

A7

21

23

22

20

21

A8

22

24

25

20

22

A9

21

23

22

20

21

Levofloxacin

27

26

28

-

-

Amphotericin B

-

-

-

23

24

 


 

Picture No.02: Antibacterial Study (In-Vitro) in Lab.

 


 

Picture No.03: Antibiotic Zone Reader

 

 

Picture No.04: Antibiotic Zone Development

 

Antibacterial activity (In-Vivo):

(Inhibitory effect ofPeel Extracts ofRed Dragon Fruit ongrowth of P. aeruginosa)

Thirty male Bal b/c mice (26-28 gm) aged between 12 and 16 weeks were used for allin vivo experiments. They were kept in a light, food and temperature-controlled room and all mice were acclimatized for at least one week prior to beginning the experiments. The dorsal back skins of the mice were shaved and ethanol (70%) was used as antiseptic for the shaved regionand then burned by using inflamed knife and then contaminated with bacteriaP.aeruginosa (1.5×108 bacteria /ml), which was the more common species in burn infection in present study. After two days of injury the inflammation, redness and suppuration region were observed and the experimental mice were randomly divided into the following groups (n = 30)

 

Group 1. untreated mice (control).

Group 2. burn mice treated with ointment composed of antibiotic (Cm).

Group 3. burn treated with ointment (concentration 50mg/ml) consisting of 0.5g of aqueous extract added to 9.5g of Vaseline.

Group 4. burn treated with ointment (concentration 100mg/ml) consisting of 1g of aqueous extract added to 9 g of Vaseline.

Group 5. burn treated with ointment (concentration 200mg/ml) consisting of 2g of aqueous extract added to 8 g of Vaseline.

Group 6. burn treated synergistically withointment Cm and 100mg/ml of aqueouscrude extract of P. granatum.

 

The treatment continued twice each dayduring 15consecutive days and thenumbers of bacteria was calculated.

 

Statistical analysis:

Results were expressed in Mean±SD andwere analyzed by one way analysis of variance (ANOVA) and post HOC analysiswas done using Tukeys using SPSS package version 16. P value< 0.05 wasconsidered to be significant.


Table No 02: Numbers of P. aeruginosa between the Mice Groups.

Duration of

Treatment per day

Group (1)

Control

Group (2)

CM

Group (3)

Aqueous extract

50mg/ml

Group (4)

Aqueous extract

100mg/ml

Group (5)

Aqueous extract

200mg/ml

Group (6)

CM+Aqueous extract

100mg/ml

3

0

145

175

165

160

125**

6

0

115

80

70*

82

7*

9

0

32

20

10

35

7*

12

0

0

2

1

5

0

15

0

0

0

0

0

0

* Significant at P ≤ 0.001

 


RESULTS AND DISCUSSION:

Antibacterial activity (In-Vitro):

The extract A1, A2, A3, A5, A8 has excellent Antibacterial activity S. aureus, the extract A1 have shown Antibacterial activity B. subtilis, while A4 show

 

Antibacterial activity against E. coil, when compared with standard Levofloxacin.

 

Antibacteria activity (In-Vivo):

The in vivo assay revealed that the treatment on mice with the different concentrations ofaqueous extract of Red Dragon Fruit peels and Cm antibiotic during 15 days and it was found to have marked effects on the number of P.aeroginosa especially the group six which represent 100mg/ml of aqueous extract and Cm antibiotic which showed more effectiveness than other groups and the number of bacteria was reduced significantly (P ≤ 0.001) from 120 to 5×106 cell/ml during the treatment days and this results due to Red Dragon Fruit Peel extract which have antibacterial activities and exhibited synergistic effects when used with antibiotic Cm as well as have wound healing properties. In addition, according to the results presented in table 3 it was noted that the group four (100mg/ml aqueous extract) significantly (P≤ 0.001) was more effective than other groups and generally the skin of the mice was healed after nine days of treatments.

 

CONCLUSIONS:

The In Vivo and In-Vitro study clearly demonstrated that the importance of Red Dragon Fruit Peelextracts in the control of burn infection resistant bacteria which are becoming a threat to human health and the aqueous extracts of peels are reported to have therapeutic properties.

 

ACKNOWLEDGEMENT:

Authors are sincerely thankful to Loknete Shri Dadapatil Pharate College of Pharmacy, Mandavgan Pharata, Shirur, Pune. (M.S.) for providing necessary facilities for work and authors also thankful to Savitribai Phule Pune University, Animal Experiments Department, Pune forproviding in-vivo activity facilities.

 

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Received on 29.09.2018         Modified on 30.10.2018

Accepted on 21.11.2018  ©A&V Publications All right reserved

Res. J. Pharmacognosy and Phytochem. 2019; 11(1): 23-26.

DOI: 10.5958/0975-4385.2019.00005.0