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Table of Contents
ORIGINAL ARTICLE
Year : 2022  |  Volume : 9  |  Issue : 2  |  Page : 37-44

Comparative evaluation of the antimicrobial activity of various concentrations of nonalcoholic extracts of crude coconut shell oil, orange peel, and mango leaf with that of xylitol on Streptococcus mutans and Candida albicans: An in vitro study


1 Professor and HOD, Department of Pediatric and Preventive Dentistry, Malabar Dental College and Research Centre, Edappal, Kerala, India
2 Reader, Department of Pediatric and Preventive Dentistry, Malabar Dental College and Research Centre, Edappal, Kerala, India
3 Professor, Department of Pediatric and Preventive Dentistry, Malabar Dental College and Research Centre, Edappal, Kerala, India
4 Senior Lecturer, Department of Pediatric and Preventive Dentistry, Malabar Dental College and Research Centre, Edappal, Kerala, India
5 Former Postgraduate Student, Department of Pediatric and Preventive Dentistry, Malabar Dental College and Research Centre, Edappal, Kerala, India

Date of Submission14-May-2022
Date of Acceptance29-May-2022
Date of Web Publication28-Jun-2022

Correspondence Address:
Dr. R Krishna Kumar
Department of Pediatric and Preventive Dentistry, Malabar Dental College and Research Centre, Edappal, Kerala
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijpcdr.ijpcdr_12_22

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  Abstract 


Background: The use of plant extracts, as well as other alternative forms of medical treatments, is enjoying great popularity since late 1990s. Scientists from all over the have found literally thousands of phytochemicals which have inhibitory effects on all types of microorganisms in vitro. Moreover, there are the reports of potential hazards and microbial resistance against the commonly used antibiotics.
Aim: The objective of this study is to evaluate the antimicrobial effect of various concentrations of nonalcoholic extracts of crude coconut shell oil, orange peel, and mango leaf with that of xylitol on Streptococcus mutans and Candida albicans.
Materials and Methods: Microbial strains were procured and revived on nutrient agar media. Coconut shell oil extract was prepared by heating ground shell in an earth pot for 3 h. Mango leaf extract was collected by powdering them using a blender and water extracts were prepared using agitation method. Orange peel extract was collected from the fresh peels of oranges which was pureed in a blender. The puree was subjected to hydrodistillation. The oil was separated. 10 g of Xylitol was dissolved in 10 ml of distilled water for base extract. 25%, 50%, 100%, and 200% dilutions of the extracts and xylitol were prepared. The susceptibility of the oral pathogens was determined by the well diffusion method. The extracts were pipetted into the wells and then incubated at 37°C overnight. After overnight incubation, the diameter of the clear zone produced was measured in cm.
Minimum inhibitory concentration (MIC) of the extracts was evaluated by the microdilution method. Serial dilutions of the extracts and xylitol were added to the wells prepared in the agar plate along with nutrient broth and culture. The plates were placed in an incubator at 37°C for 24 h. The absence of turbidity in the wells was recorded as MIC.
Results: Highest zone of inhibition was recorded for coconut shell extract against both the test organisms followed by Xylitol. Mango leaf extract and orange peel extract could not demonstrate statistically significant results. The inhibitory effect increased with increasing concentration. MIC of coconut shell extract against C. albicans and S. mutans was found out to be 37.5 mg and 25 mg, respectively.
Conclusion: Based on the results of the study, it may be concluded that antibacterial and antifungal effect of nonalcoholic extracts of crude coconut shell extract is more when compared to mango leaf, orange peel, and xylitol. Mango leaf and orange peel extracts showed minimum activity.

Keywords: Candida albicans, coconut shell, inhibitory effect, mango leaf, nonalcoholic extract, orange peel, Streptococcus mutans, xylitol


How to cite this article:
Peter J, Kumar R K, Vijai S, Augustin M, Anaswara M S, Ajaykumar A. Comparative evaluation of the antimicrobial activity of various concentrations of nonalcoholic extracts of crude coconut shell oil, orange peel, and mango leaf with that of xylitol on Streptococcus mutans and Candida albicans: An in vitro study. Int J Prev Clin Dent Res 2022;9:37-44

How to cite this URL:
Peter J, Kumar R K, Vijai S, Augustin M, Anaswara M S, Ajaykumar A. Comparative evaluation of the antimicrobial activity of various concentrations of nonalcoholic extracts of crude coconut shell oil, orange peel, and mango leaf with that of xylitol on Streptococcus mutans and Candida albicans: An in vitro study. Int J Prev Clin Dent Res [serial online] 2022 [cited 2022 Aug 16];9:37-44. Available from: https://www.ijpcdr.org/text.asp?2022/9/2/37/348707




  Introduction Top


When it comes to oral diseases, dental caries is the first and foremost ailment that depicts the picture. Even though the human race is equipped with several preventive and treatment measures, the fact remains that caries is spreading like a wild fire among the human populations. It can be implied that even the latest developments in the field of caries prevention are not able to prevent the disease and its eradication seems to be a distant dream.[1]

Antimicrobials have been the mainstay in caries prevention, but the increasing resistance of the microbes involved in the disease, a global call for the search of alternative preventive methods is on the rise, also the economic burden of the disease has to be relieved from the developing nations.[2],[3] Bacterial resistance to most of the antibiotics commonly used to treat oral infections (penicillins and cephalosporins, erythromycin, tetracycline and derivatives, and metronidazole) has been documented by P. Bidault, F. Chandad, and D. Grenier.[4] Cetylpyridinium chloride, chlorhexidine, amine fluorides or products containing such agents, which are very useful adjuncts to the chemomechanical prevention are reported to exhibit toxicity, cause staining of teeth.[5],[6] Ethanol (commonly found in mouthwashes) has antiseptic prorerties but also has been linked to oral cancer.[7],[8]

Green dentistry is eco-friendly dentistry which encompasses a simultaneous devotion to sustainability, prevention, precaution, and a minimally invasive patient-centric as well as global-centric treatment philosophy.[9] Several studies have been published demonstrating the antimicrobial effects of various natural products. To our best knowledge, there are no studies which compare the antibacterial and antifungal effects of different concentrations of nonalcoholic extracts of crude coconut shell oil, orange peel and mango leaf with that of xylitol on Streptococcus mutans and Candida albicans.


  Materials and Methods Top


This study was conducted from the Department of Pedodontics and Preventive Dentistry, Malabar Dental College and Research Center, Edappal in collaboration with the Unibiosys Labs, Kochi.

Armamentarium

For the purpose of the conducting the study, the following armamentarium was used:

  1. S. mutans MTCC 890
  2. C albicans MTCC 3017
  3. Orange (Citrus sinensis) peel-250 g
  4. Mango (Mangifera indica) leaves-250 g
  5. Dried coconut (Cocos nucifera) shell-250 g
  6. Xylitol (So Sweet, Herboveda, India)-250 g
  7. Hydrodistillation (Clevenger) apparatus
  8. Beaker
  9. Earthen pot
  10. Gas stove
  11. Petridishes
  12. Mueller Hilton Agar
  13. Nutrient Agar Medium
  14. Saborauds Agar Medium
  15. Whatman filter paper
  16. Microtitre plate
  17. Optical density reader
  18. Gloves
  19. Mask.


Prepration of crude coconut shell extract

Clean and dry Coconut (Cocos nucifera) shell was powdered finely and 250 g of the powder was put in an earthen pot as described by Dorathy et al.[10] The assembly was heated in indirect heat over burner. At the end of 3 h, vapors of the crude coconut shell oil were deposited on the inside of the foil. The extract was wiped off the sheet and stored in for subsequent preparation of various dilutions. To 1.0 g of the material 10 ml of distilled water was added and the mixture was boiled and allowed to cool. The insoluble matter was removed by centrifugation. The procedure was repeated three times as coconut shell extract is oily in nature. The water in the filtrate was evaporated to complete dryness using a standard Buchi rotary-evaporator. Dry extracts were obtained which were re-suspended in 5 ml distilled water. In order to determine the real concentration of each extract, 1 ml of the homogenization of the extract was completely oven-dried and then weighed to determine amount of extract per ml of final solution.[11] The extract was prepared at the strength of 500 mg/ml. Various dilutions were subsequently prepared by adding different volumes of distilled water to 1 ml of the base extract, i.e., 0.25 ml for 25%, 0.50 ml for 50%, 1 ml for 100%, and 2 ml for 200% dilutions.

Preparation of mango leaf extract

Mango (Mangifera indica) leaves available locally were identified at Kerala Agricultural University and used for the study. The leaves were washed and shade dried, and after grinding in an electric grinder, 250 g of the powder were soaked in 250 ml of distilled water. The beaker was covered. Agitation extraction was prepared by mixing intermittently as described by Anand et al. in 2015[11] and Madhuri et al. in 2016.[12] The filtrate was collected in a jar and stored in the refrigerator for further use. Using a standard Buchi rotary-evaporator, the water was completely evaporated to yield dry extract. Dry extract was re-suspended in 5 ml distilled water. In order to determine the real concentration of each extract, 1 ml of the homogenized extract was completely oven-dried and then weighed to determine amount of extract per ml of final solution.[13] The extract obtained had a concentration of 500 mg/ml. The various dilutions were subsequently prepared by adding distilled water to 1 ml of the base extract, i.e., 0.25 ml for 25%, 0.50 ml for 50%, 1 ml for 100%, and 2 ml for 200% dilutions.

Preparation of orange peel extract

Fresh oranges (Citrus sinensis) were purchased from local fruit vendor and were identified at Kerala Agricultural University, Mannuthy. Peel was removed and 250 g of the peel was pulverized to puree in a blender. The peel puree was subjected to hydro distillation using Clevenger Apparatus in a 1 L round bottom flask over a heating mantle with antibumping granules. As the distillation progressed the oil and water layer separated in a collecting flask. The oil was separated using a dropper and was stored in amber colored vials according to the method described by Hasija et al. in 2015[14] and stored in the refrigerator for further use. 25 ml of extract was collected at the end of 6 h. The method described by Bussman et al.[13] was employed to prepare the dry extract which was later suspended in 5 ml of distilled water to yield a concentration of 500 mg/ml. The various dilutions were subsequently prepared by adding distilled water to 1 ml of the base extract, i.e., 0.25 ml for 25%, 0.50 ml for 50%, 1 ml for 100%, and 2 ml for 200% dilutions.

Preparation of xylitol solution

The Xylitol base solution was made by mixing 10 g xylitol powder and 10 ml distilled water until homogeneous according to the method described by Auerkari et al.[14] From this base solution, 25%, 50%, 100%, and 200% diluted Xylitol solutions were prepared separately by adding distilled water in the amounts of 0.25 ml, 0.50 ml, and 1 ml, respectively, to 1 ml base solution. The concentration of the solution was 100 mg/ml.

Microbial strain procurement

Freeze dried S. mutans MTCC890 and C albicans MTCC3170 were obtained from Microbial Type Cell Culture and Gene Bank, Institute of Microbial Technology, Chandigarh. S. mutans was revived on Nutrient Agar Medium, C albicans was revived on Saborauds Agar Medium according to. The plates were incubated at 37 degrees for 24 h.

Determination of inhibitory effect

The inhibitory effect of the various extracts was measured using the well diffusion method. Using aseptic techniques, a single pure colony was transferred into a 10 ml of nutrient broth, capped and placed in incubator overnight at 37°C. After 18 h of incubation, using aseptic preparation, turbidity of microbial suspensions was calculated and adjusted using McFarland standards as a reference.

Microbial suspensions were streaked onto agar plates containing Meuller Hilton Agar. Five wells were prepared on each agar plate using a sterile stainless-steel template. Using a sterile micropipette 125 μl of specific concentration of the extracts was dispensed into each well. This was done in triplicate for every concentration to overcome any inadvertent technical errors.

Following 48 h of incubation at 37°C, zones of inhibition (that is, locations where no growth of bacteria was present) were examined around the wells. These appeared as a clear, circular halo surrounding the wells. The diameters of the inhibition zones were measured using a Hi Antibiotic Zone Scale (Hi Media Laboratories, Mumbai, India) [Figure 1] and [Figure 2].
Figure 1: Revival of C. albicans. C. albicans: Candida albicans

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Figure 2: Revival of S. mutans. S. mutans: Streptococcus mutans

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Determination of minimum inhibitory concentration

To determine minimum inhibitory concentration (MIC), various dilutions of the extracts were taken in sterile microtiter plate using nutrient broth as diluents and a tube containing nutrient broth was taken as control. 50 μl of the standard culture inoculum was added to each tube, except the control tube. All tubes were incubated at 37°C for 24 h and then examined for growth by observing turbidity. 10 μl of microbial culture was pipetted from the mixture obtained in the determination of MIC tubes, which did not show any growth and sub-cultured onto Muller Hinton agar and incubated at 37°C for 24 h. After incubation, the concentration at which there was no single colony of bacteria was taken as MIC.


  Results Top


The results were analyzed using the SPSS software version 22 IBM, Chicago, Illinois, United States. Kruskal–Wallis test was applied setting concentration as the grouping variable. The zones of inhibitions of crude Coconut shell extract against C. albicans and S. mutans in cm are shown in [Table 1]. In case of C albicans, the mean zones of inhibition for 0%, 25%, 50%, 100%, and 200% of dilutions are 1.400 ± 0.200, 1.266 ± 0.152, 1.066 ± 0.057, 1.000 ± 0.000, 1.000 ± 0.000, respectively, whereas, the zones of inhibition against S. mutans are 1.266 ± 0.115, 1.133 ± 0.057, 1.033 ± 0.057, 1.000 ± 0.000, and 1.000 ± 0.000, respectively.
Table 1: Comparison of zones of inhibition (cm) of coconut shell extract on Candida albicans and Streptococcus mutans

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The zones of inhibitions of Mango leaf extract against C albicans and S. mutans in cm are shown in [Table 2]. In case of C albicans, the mean zones of inhibition are 1.000 at all the dilutions, i.e., 0%, 25%, 50%, 100%, and 200% dilutions. The same values were obtained against S. mutans also, i.e., 1.000 at the respective dilutions.
Table 2: Comparison of zones of inhibition (cm) of mango leaf extract on Candida albicans and Streptococcus mutans

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The zones of inhibition of orange peel extract are shown in [Table 3]. The extract demonstrated 1.066 ± 0.115, 1.033 ± 0.057, 1.000 ± 0.000, 1.000 ± 0.000, 1.000 ± 0.000 at the respective dilutions in case of C. albicans. However, when it came to S. mutans, the values were 1.000 at all the dilutions tested.
Table 3: Comparison of zones of inhibition (cm) of orange peel extract on Candida albicans and Streptococcus mutans

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In case of Xylitol, the values are depicted in [Table 4]. At the respective dilutions of 0%, 25%, 50%, 100%, and 200%, the zones of inhibition were 1.066 ± 0.115, 1.033 ± 0.057, 1.000 ± 0.000, 1.000 ± 0.000, and 1.000 ± 0.000 against C. albicans. It demonstrated the same mean zones of inhibition as mango leaf and orange peel extract against S. mutans at 25, 50, 100, and 200, i.e., 1.000 ± 0.000 dilutions, respectively, but it demonstrated 1.200 ± 0.173 at 0%.
Table 4: Comparison of zones of inhibition (cm) of xylitol on Candida albicans and Streptococcus mutans

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The MIC s of various extracts are shown in the [Table 5]. Mango leaf extract and orange peel extract failed to show any MIC against the test organisms. The MIC demonstrated by xylitol was 50 mg against S. mutans and C. albicans. However, the MIC shown by coconut shell extract was the least when compared with the other test extracts. The value obtained was 37.5 mg for C. albicans and 25 mg for S. mutans.
Table 5: Minimum inhibitory concentration

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In coconut shell extract group, the P = 0.017 in C. albicans group and 0.015 in S. mutans group. This indicates that the inhibitory effect exerted by coconut shell extract on the test organisms, i.e., C. albicans and S. mutans is statistically significant. However, in mango leaf extract group, the P = 1.0 against both the organisms. This indicates that the inhibitory effect exerted by mango leaf extract on the test organisms, i.e., C. albicans and S. mutans is not significant statistically. Furthermore, the P = 0.519 in C. albicans group and 1.00 in S. mutans group in case of Orange peel extract. This indicates that the inhibitory effect exerted by orange peel extract on the test organisms, i.e., C. albicans and S. mutans is not significant statistically In Xylitol solution group, the P = 0.519 in C. albicans group and 0.030 in S. mutans group. This indicates that the inhibitory effect exerted by Xylitol solution on C. albicans is not significant statistically, but against S. mutans, the action is statistically significant [Figure 3] and [Figure 4].
Figure 3: Zone of inhibition on C. albicans- (a) Coconut shell. (b) Mango leaf. (c) Orange peel. (d) Xylitol. C. albicans: Candida albicans

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Figure 4: Zone of inhibition on S mutans in triplicates- (a) Coconut shell extract. (b) Mango leaf extract. (c) Orange peel extract. (d) Xylitol. S. mutans: Streptococcus mutans

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  Discussion Top


By finding ways to maintain ecological stability in the oral cavity, we can prevent S. mutans from ever attaining the strength necessary to cause harm in accordance with Loesche's specific plaque hypothesis.[15],[16]

C. albicans is by far the most commonly detected fungal organism on human mucosal surfaces. It is an opportunistic pathogen that lives as a benign commensal organism in the mouths of healthy individuals, especially younger children. One characteristic of C. albicans that makes it a successful opportunistic pathogen is its ability to adapt and proliferate in a broad range of host environments.[17] Studies have shown the ability of C. albicans in dissolving hydroxyapatite[18] and causing caries in vivo.[19] Its link with early childhood caries has been demonstrated by Falsetta et al.,[20] by exhibiting synergistic interactions with S. mutans and codetection with S. mutans in plaques obtained from early childhood caries patients.[21]

Ever-changing dynamic conditions in the mouth holds the ability to hoard myriads of microorganisms, of which only S. mutans and C. albicans are the persistently found in caries active individuals. The alcoholic base in the various prophylactic agents used in dentistry has shown its carcinogenic potential. Especially in pediatric dentistry, we should aim to achieve a nonalcoholic therapeutic agent where there is always a chance of the child patient ingesting these chemical based mouthwashes. Hence, this study was directed to understand the various concentrations of natural nonalcoholic extracts of coconut shell, mango leaf, and Orange peel on these pathogens when compared with Xylitol. To our best knowledge, there has not been any comparison between the test specimens that are used in this study.

The results clearly demonstrate that higher the concentration the more potent is the solution. The serial dilutions have demonstrated less potency, 200% dilution being least potent and nonsignificant in accordance with the previous studies performed.[11],[22]

In case of coconut shell, the mean zones of inhibition for 0%, 25%, 50%, 100%, 200% of dilutions are 1.400 ± 0.200, 1.266 ± 0.152, 1.066 ± 0.057, 1.000 ± 0.000, and 1.000 ± 0.000, respectively, against C. albicans. In case of mango leaf extract, the mean zones of inhibition are 1.000 at all the dilutions. Orange peel extract demonstrated 1.066 ± 0.115, 1.033 ± 0.057, 1.000 ± 0.000, 1.000 ± 0.000, and 1.000 ± 0.000 at the respective dilutions. Xylitol demonstrated same values as orange-peel extract. On comparison of the zones of inhibitions of mango leaf extract, orange peel extract and Xylitol on C. albicans, it is seen that the results are not statistically significant as the P values are <0.05. However, in the case of coconut shell oil, the P = (0.017) is lesser than 0.05 which indicates the zones of inhibition produced by coconut shell extract on C. albicans are significant.

When it came to S. mutans, the values obtained were 1.266 ± 0.115, 1.133 ± 0.057, 1.033 ± 0.057, 1.000 ± 0.000, and 1.000 ± 0.000, respectively, at the dilutions of 0%, 25%, 50%, 100% and 200% dilution. In case of mango leaf extract, the zones are 1.000 at all the dilutions. Orange peel extract also demonstrated 1.00 ± 0.00 at all the respective dilutions. Xylitol demonstrated the same mean zones of inhibition as mango leaf and orange peel extract at 25%, 50%, 100%, and 200% dilutions, respectively, but it demonstrated 1.200 ± 0.173 at 0%. On comparison of the zones of inhibitions of mango leaf extract and orange peel extract on S. mutans, it is seen that the results are not statistically significant as the P values are <0.05, i.e., 1.000 in both cases. However, in the case of coconut shell extract and Xylitol, the P = (0.015 and 0.030 respectively) is lesser than 0.05 which indicates the zones of inhibition produced by coconut shell extract on S. mutans are significant.

The present study demonstrated greater inhibitory effect of coconut shell extract against the test organisms when compared to mango leaf extract, orange peel extract, and Xylitol. To our best knowledge, the comparison of coconut shell extract has not been done against the aforementioned extracts and solutions. Increasing dilutions have demonstrated progressively decreasing inhibitory effects.

The studies revealed that the dry coconut shells are the source of certain organic materials such as polyphenols and organic acids.[23] This material also has great potential and may be a good source for future antibiotic as active biocomponents such as tocopherol, palmitoleyl alcohol, cycloartanol, and β-sitosterol have been identified in its extract.[24] However, the activity against the tested microorganisms in the aforementioned study was found in the presence of various organic solvents, which themselves can have an effect on the tested pathogens.

The aqueous extract of coconut shell oil contains alkaloids, carbohydrates, saponin, phenols, tanins, terpenoids, proteins, oxalate, carboxylic acid, quinones, and glycosides.[10] Dihydroxy benzoic acid, methoxybenzoic acid, and 150 hydroxyl benzoic acid are present as minor acidic components.[25] The inherent composition of coconut shell extract could be the reason for its high antimicrobial effect when compared to other extracts used in the study. The results of the study by S. Dorathy et al. indicate that the zone of inhibition increased when increasing the volumes of the extracts which was in accordance to our study. Ethanol and petroleum ether extract showed maximum inhibitory activity against Epidermophyton and C. albicans in that study.[10]

Mango leaf extract was not able to exert statistically significant inhibitory effects on the test organisms which was in contradiction with the study performed by Bhatt et al.[26] who found significant reductions of oral Streptococcal counts after using mango leaf mouthwash. The difference can be accounted to the ethanolic solution which was used to prepare the mouth wash. The present study used aqueous extract of mango leaf. Thus, the base was nonalcoholic.

Mangifera indica contains the pharmacologically and medicinally important chemical mangiferin, which is a polyphenolic antioxidant. It has cytotoxic and apoptotic effects.[27] Ethanolic extract show greater efficacy in inhibiting the growth of S. mutans as compared to aqueous extracts which are effective at only at higher concentrations.[28] It was noted in a study done by Dikonketso et al.[29] that mango leaf extract had an ability to target the expression of the gftB gene, thereby targeting bacterial attachment. However, in that study ethanol was used to prepare the extract.

Inhibitory effects on test organisms was statistically not significant with the used of orange peel extract, even though it was reported to have high antimicrobial potency.[30] This variation can be due be the effect of the base which is used in the extract prepared. The inhibitory activity of the orange peel oil may be a cumulative effect of D-limonene and some other unidentified components or flavonoids. D-limonene is a terpene found in orange peel which constitutes more than 90% of the phytochemicals present in the peel extract.[31] Unal et al.[32] also demonstrated the antifungal and inhibitory effect of D-limonene on a variety of yeast strains and the inhibitory affect was attributed to attributed to a disruption of the cellular membrane which can cause the cellular contents to leak out of the cell. Disruption of H + and K+ transport also suggested as mechanism of action of D-limonene.

Nevertheless, the results of the present study was similar to the reports by revealing less activity of aqueous extracts against C. albicans when compared to chloroform and ethanol extracts. Cowan[32],[33] has reported that most active components are not water soluble indicating that there are some active ingredients in plant extracts which have high antimicrobial effect but which would not be released except when orange a particular solvent is used. He also mentioned that most of the antibiotic compounds already identified in plants are reportedly aromatic or saturated organic molecules are soluble in organic solvents.

In our study, the already proven effects of xylitol against S. mutans and C. albicans were reinforced. But in comparison with Coconut shell extract, Xylitol proved to be less effective. Pizzo et al.[34] found that xylitol reduces adherence of Candida species to epithelial surfaces, therefore, resulting in an inhibitory effect. Radmerikhi et al.[35] concluded that the consumption of 5% xylitol reduced S. mutans and Lactobacillus acidophilus growth and by an increase in concentration of xylitol, the inhibitory effect increased. They confirmed that even in low concentrations, Xylitol has an inhibitory effect against main oral cavity inducer bacteria.


  Conclusion Top


Based on the results of the study it may be concluded that antibacterial and antifungal effect of nonalcoholic extracts of crude coconut shell extract is more when compared to mango leaf, orange peel and xylitol. Mango leaf and orange peel extracts showed minimum activity. The antimicrobial effect increased with increasing concentration. Toxicity of these materials should be analyzed to determine the safety. It can be concluded that the coconut shell extracts which can be prepared at home without any technique sensitivity and can provide optimal protection against S. mutans and C. albicans in underprivileged populations. With the help of this study, the aim of introducing a novel, economic prophylactic agent has been established.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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