|Year : 2020 | Volume
| Issue : 1 | Page : 72-77
Evaluation of the effectiveness of probiotics consumption on salivary Streptococcus Mutans counts in mentally retarded children: An Ex Vivo study
Priti Murarka, Nilima Thosar, Sudhindra Baliga, Nilesh Rathi, Suruchi Gupta
Department of Pedodontics and Preventive Dentistry, Sharad Pawar Dental College, Datta Meghe Institute of Medical Sciences (Deemed University), Wardha, Maharashtra, India
|Date of Submission||21-Mar-2020|
|Date of Decision||27-Mar-2020|
|Date of Acceptance||29-Mar-2020|
|Date of Web Publication||13-Oct-2020|
Dr. Suruchi Gupta
Department of Pedodontics and Preventive Dentistry, Sharad Pawar Dental College, Datta Meghe Institute of Medical Sciences (Deemed University), Sawangi (Meghe), Wardha - 442 004, Maharashtra
Source of Support: None, Conflict of Interest: None
Background: Probiotics are those live micro-organisms which, when administered in adequate amounts, confer a health benefit to the host. Aim: The purpose of the study was to evaluate the effectiveness of probiotics consumption on levels of salivary Streptococcus mutans in mentally retarded children. Materials and Method: The study was undertaken with 30 institutionalized healthy children and 30 institutionalized mentally retarded children of 4-14 years of age group. The children were randomly divided into two groups, each comprising 15 children; Study group I: Mentally retarded children consuming probiotics, Study group II: Mentally retarded children not consuming probiotics, Control group I: healthy children consuming probiotics, Control group II: healthy children not consuming probiotics. The children consumed probiotics for 4 weeks. Streptococcus mutans/ml of saliva was collected at baseline, after 2nd week and 4th week. Results: A statistically significant reduction (P<0.05) in streptococcus mutans counts was recorded in probiotics ingestion groups. Conclusion: A positive correlation of probiotics with mean streptococcus counts was observed with both the groups, suggestive of probiotics role in caries prevention.
Keywords: Caries, mentally retarded children, probiotics
|How to cite this article:|
Murarka P, Thosar N, Baliga S, Rathi N, Gupta S. Evaluation of the effectiveness of probiotics consumption on salivary Streptococcus Mutans counts in mentally retarded children: An Ex Vivo study. J Datta Meghe Inst Med Sci Univ 2020;15:72-7
|How to cite this URL:|
Murarka P, Thosar N, Baliga S, Rathi N, Gupta S. Evaluation of the effectiveness of probiotics consumption on salivary Streptococcus Mutans counts in mentally retarded children: An Ex Vivo study. J Datta Meghe Inst Med Sci Univ [serial online] 2020 [cited 2020 Oct 31];15:72-7. Available from: http://www.journaldmims.com/text.asp?2020/15/1/72/297990
| Introduction|| |
“Dental caries is considered to be an irreversible microbial disease of the calcified tissues of the teeth which are characterized by demineralization of the inorganic portion and destruction of the organic substance of the tooth, which often leads to cavitation.” Main etiological factors for dental caries are microorganisms, carbohydrates (sucrose and fructose), susceptible tooth, and the time.
Streptococcus mutans as an effective cariogenic microbe has been seen in studies.S. mutans has been identified as the principal cariogenic bacterium for caries initiation, while Streptococcus sobrinus ught to enhance caries initiation, progression, and development., Virulence properties of S. mutans which are important for their proliferation and establishment are acidogenicity and acidurance which allows the organisms to tolerate low pH levels and contributes significantly to their cariogenic potential.,
Saliva has a vital role in upholding the integrity of the oral structures. Saliva can affect the occurrence of dental caries in many ways, one of them being by a drop in enamel solubility with a unceasing supply of minerals, chiefly calcium, and by buffering and antibacterial activity.
Probiotics, literally meaning “for life,” in which microorganism prove to exert health-promoting influences in humans and animals. Bacterial replacement therapy in the form of probiotics seems to be a natural way to maintain health and protect oral tissues from the disease. The most common probiotic bacteria belong to the lactobacilli and bifidobacteria genera, but few strains of streptococci have also been investigated. Within dentistry, studies with Lactobacillus rhamnosous GG, Lactobacillus reuteri have potential in interacting with S. mutans by reducing the number of this cariogenic bacteria, thus suggesting a role of probiotics in caries prophylaxis.,,
“Mental retardation has been defined by the American Association of Mental Deficiency as a deficiency in theoretical intelligence that is congenital or acquired in early life.” Dental caries is the most prevalent disease among mentally retarded children worldwide and “dental treatment is the greatest unattended health need among the disabled.” The most important reasons may be inadequate recall systems, practical difficulties which a clinician come across are treatment sessions, socioeconomic status, underestimation of need for the treatment or severity of pain, communication problems, and bad cooperation.,,,, The effects of disabling conditions are several and varied, but out of them, most common effects is the inability of the individual to maintain dental health.,
“Taking this into account, present study was planned to evaluate the effectiveness of probiotics consumption on levels of salivary S. mutans in mentally retarded children.”
| Materials and Methods|| |
“Institutional Ethical Committee approval was taken before starting the study. An informed consent of parents/legal guardians was obtained prior to the study.” Thirty institutionalized healthy children and thirty institutionalized mentally retarded children of 4–14 years of age group were included in the study. Habitual consumers of xylitol chewing gums or children on systemic antibiotics or on topical fluoride treatment were excluded from the study.
The study group consisted of 30 mentally retarded children which were further divided into two subgroups of 15 children each. Study Group I consisted of 15 institutionalized mentally retarded children consuming probiotics, study Group II consisted of 15 institutionalized mentally retarded children not consuming probiotics in the same age group. Control group consisted of 30 normal healthy children which were further divided into two subgroups of 15 children each. Control Group I consisted of 15 institutionalized normal healthy children consuming probiotics, control Group II consisted of 15 institutionalized normal healthy children not consuming probiotics in the same age group. Probiotics were consumed for 4 weeks.
- Study Group I: Institutionalized mentally retarded children consuming probiotics (n = 15)
- Study Group II: Institutionalized mentally retarded children not consuming probiotics (n = 15)
- Control Group I: Institutionalized normal healthy children consuming probiotics (n = 15)
- Control Group II: Institutionalized normal healthy children not consuming probiotics (n = 15).
Intraoral examination was carried out by making children sit in the upright position and under proper illumination using diagnostic instruments. Oral health status was evaluated using the WHO oral health assessment form for children (2013). Oral hygiene status was recorded using simplified oral hygiene index given by Greene and Vermilion (1964).
Saliva samples were collected at baseline [Figure 1], after the 2nd week and after 4th week of intervention period. Saliva sampling was performed between 9 and 10 a. m. in the morning to minimize the effect of diurnal variation. Paraffin stimulated whole saliva was collected for 2 min in a saliva collection cup. Collected saliva was transferred to the microbiology laboratory within 3 h. Saliva samples were vortexed and serially diluted in ten-fold steps in 0.05M phosphate buffer.
Aliquots of 100 μl of the appropriate dilutions were cultured into selective media for MS, which has mitis salivarius bacitracin (MSB) agar [Figure 2]. All plates were cultivated at 37°C in a micro-aerophilic environment in 5% CO2 for 48 h. Digital colony counter was used for calculating and Identification of the colonies was done based on their morphology. Blinding of the laboratory technicians and clinicians was done to prevent bias regarding allocation of study individuals. The obtained data were subjected to statistical analysis. Data analysis was performed by using descriptive and inferential statistics both using Mann–Whitney test. “The software used in the analysis was SPSS (Statistical Package for Social Sciences) Version 24.0 (IBM Corp, Chicago, USA)and P < 0.05 was considered as level of significance.”
| Results|| |
A total of 30 institutionalized mentally retarded children and 30 institutionalized healthy children participated in the study.
When gender-wise distribution was assessed in the study group and control group, it was observed that out of 30 mentally retarded children, 16 were male and 14 were female, whereas in healthy group, 13 were male and 17 were female.
[Table 1] shows demographic parameters of the children such as age, dental caries along with oral hygiene status. It was observed that mean age in mentally retarded and healthy children was almost equal, i. e; 11.23 ± 0.35 and 11.30 ± 0.53, respectively. Similarly, the mean caries status was also similar in both the groups, i.e.; 0.33 ± 0.55 and 0.33 ± 0.61, respectively. Mean oral hygiene status was less in mentally retarded group (0.77 ± 0.68) as compared to healthy group (1.07 ± 0.78) [Table 1].
|Table 1: Comparison of age, dental caries and oral hygiene status in mentally retarded and healthy children|
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[Table 2] shows, at the 2nd week of follow-up, the mean S. mutans count remained almost same 103.22 ± 29.28 in children without probiotics use, whereas in children who used probiotics, it decreased to 92.53 ± 25.39. Statistically, the difference between both the groups was found to be statistically significant (0.042, P < 0.05). Similar findings were seen at 4th week of follow-up with no change in count in children without probiotics group (105.65 ± 25.86) and reduced levels in children with probiotic usage (81.33 ± 25.48) which was again statistically significant (0.036, P < 0.05).
|Table 2: Comparison of mean Streptococcus mutans n mentally retarded children using probiotics and not using probiotics|
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As shown in [Table 3], at the 2nd week of follow-up, the mean S. mutans count remained almost same, i. e, 112.49 ± 23.35 in children without probiotics use, whereas in children who used probiotics it decreased to 88.73 ± 36.98. Statistically, the difference between both the groups was found to be statistically significant (0.012, P < 0.05). Similar findings were seen at the 4th week follow-up with no change in counts in healthy children without probiotics group, i. e, 115.92 ± 29.56 and reduced levels in children with probiotic usage was 74.67 ± 24.84 which was again statistically significant (0.03, P < 0.05).
|Table 3: Comparison of mean streptococcus mutans counts in healthy children using probiotics and not using probiotics by using Mann-Whitney test|
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No significant differences were observed in mentally retarded and healthy children not consuming probiotics, respectively [Table 2] and [Table 3].
From [Table 4], it was observed that at baseline, mean S. mutans count in children who used probiotics was 112.33 ± 42.16 in healthy group and 102.20 ± 24.11 in mentally retarded group. At the 2nd week of follow-up, this count reduced in both the groups, in healthy children, it came down to 88.73 ± 40.36, whereas in mentally retarded children, it decreased to 92.53 ± 25.39. Similar findings were seen at the 4th week follow-up with decline in the mean S. mutans count in both healthy and mentally retarded children and none of these differences were found to be statistically significant (0.567, P > 0.05).
|Table 4: Comparison of children using probiotics in healthy and mentally retarded children|
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| Discussion|| |
Dental caries is considered to be one of the most common chronic infectious diseases in the world.,. Inequity in the microbial flora of the oral cavity aids the caries causing microbes to increase in the oral cavity as the pH drops due to the acid production.
S. mutans, are the principal cariogenic microorganism responsible for dental caries. The high tolerance of Streptoccocus mutans to environmental acidification was found to be notable, due to various factors such as DNA repair mechanisms, a broader range of endurance from intracellular acidification, higher activity of H+-ATPases at low pH and a range of other mechanisms. In addition to these, the production of extracellular polysaccharides was also considered as an important contributor to S. mutans pathogenicity., From all these studies, S. mutans is called as the “arch-villain” in causation of dental caries.,
Dental ailments are seen even with a minor shift in native atmosphere, which stimulates the probable pathogens to gain competitive benefit under suitable conditions. Probiotics may be used to cure such dental diseases.
Probiotics are the whole bacterial replacement therapy which may be very effective in removing the dangerous pathogens from the oral cavity. An essential condition for a microorganism to act as a probiotic for oral health benefit is its capacity to adhere and to colonize various surfaces of the oral cavity. Probiotic bacteria can act through various mechanisms: They prevent cellular adhesion and invasion of pathogenic bacteria, the intestinal environment by lowering the pH level as a result of fermentation products and they interact and modulate the local and systemic inflammatory immune response., Hence, in the present study, interaction of probiotics on the S. mutans has been evaluated.
In the present study, study group was mentally retarded children as there is no literature available explaining the correlation of probiotics and S. mutans levels among these children. However, a study conducted by Sujlana et al. concluded that the use of visual pedagogy coupled with probiotic mouth rinsing may improve the periodontal status of children with hearing impairment. Various other studies done by Barone et al., Szajewska and Mrukowicz, Huang et al. explained the benefits associated with probiotics on diarrhea among these children.
Besides mentally retarded children, there are also certain other groups of children which are also associated with high levels of S. mutans counts. Those are the children with higher plaque levels as in chronic gingivitis or children undergoing fixed orthodontic treatment. A study carried out by Dhawan and Dhawan in 2013, investigated the effect of probiotic formulation on the plaque and S. mutans levels in chronic gingivitis patients over 2 weeks intervention period and he found that there was significant reduction in S. mutans count and plaque levels after probiotic consumption. Thus the conclusion of the study was probiotic could be valuable in the enhancing of oral health in such patients.
In the present study, it was observed that mentally retarded children who consumed probiotics for 2 weeks, their S. mutans levels were decreased to 92.53 ± 25.39 which was statistically significant (0.042, P < 0.05) which further reduced to 81.33 ± 25.48 after 4 weeks which was again statistically significant (0.036, P < 0.05). Similar findings were seen with the normal healthy children control group, who consumed probiotics for 2 weeks. Their S. mutans levels also decreased to 88.73 ± 36.98 which was statistically significant (0.012, P < 0.05), which further reduced to 74.67 ± 24.84 after 4 weeks which was again statistically significant (0.03, P < 0.05). This was in accordance with the study carried out by Caglar et al., discovered that short-term (3 weeks) regular consumption of lactobacilli-derived probiotics supplied in the form of prepared straws or lozenges diminishes the levels of salivary mutans streptococci in young adults.
However, similar results were obtained by Caglar et al. when children consumed ice-cream containing Bifidobacterium lactis 10 days each and other 10 days they consumed ice-cream without viable bacteria. Even then, a reduction in levels of mutans streptococci was observed after intake of the control ice-cream.
A large variety of organisms can be categorized as probiotics. Lactobacillus and Bifidobacterium genera are the most widely used strains of probiotics.
In the present study, composition of the probiotics used are Bifidobacterium longum, Lactobacillus rhamnosus and Lactobacillus acidophilus. Several other formulations of probiotics, have also been tried and tested in different studies given in the literature, Caglar et al. used L. reuteri ATCC 55730; Ritthagol et al. and Teanpaisan and Piwat used Lactobacillus paracasei SD1; Yousuf et al. used Lactobacillus acidophilus, Lactobacillus plantarum, B. longum, Bifidobacterium bifidum.
In the present study, probiotics used were in the form of powder in sachet. Dairy foods incorporated with probiotics are the natural means of oral administration and can be conveniently implemented in nutritive regime. Several studies have been carried out to study the effectiveness of probiotics in different form of dairy products like Siddiqui et al. used probiotic milk, Nagarajappa et al. used chocobar ice-cream containing probiotics. Bhalla et al. used probiotic curd and he found that pathogenic microorganisms can be displaced by probiotic bacteria. Hence, the author concluded that the use of probiotic products can be used for the prevention of enamel demineralization. However, Mahantesha et al. compared the effectiveness of probiotic ice cream and drink and concluded that the probiotic organisms definitely have a role in reducing the salivary S. mutans level but, ice cream would be a better choice than drink.
In the present study, saliva samples of both the groups, i.e., study group and control group were collected before and after the intervention period. Baseline evaluation of salivary S. mutans was done MSB agar. Recognition and calculation of the mutans colonies was done using a digital colony counter. Similar method was adapted in other studies as well, by Jindal et al.; Chinnappa et al.; Bhalla et al.; Siddiqui et al. Another method available to enumerate levels of S. mutans is chair-side kits, which was used in studies done by Caglar et al.; Cildir et al.; Dilsah et al.
| Conclusion|| |
- Higher levels of mean streptococcus counts was observed in institutionalized mentally retarded children not consuming probiotics when compared with mentally retarded children consuming probiotics after 2 weeks and 4 weeks of intervention period
- Higher levels of mean streptococcus counts was observed in institutionalized healthy children not consuming probiotics when compared with healthy children consuming probiotics after 2 weeks and 4 weeks of intervention period
- A positive correlation of probiotics with the mean streptococcus counts was observed with both the groups, suggestive of role of probiotics in caries prevention
- A negative correlation of probiotics with the mean S. mutans counts was observed when comparison of both the groups.
Declaration of patient consent
The authors certify that they have obtained all appropriate patient consent forms. In the form the patient(s) has/have given his/her/their consent for his/her/their images and other clinical information to be reported in the journal. The patients understand that their names and initials will not be published and due efforts will be made to conceal their identity, but anonymity cannot be guaranteed.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3], [Table 4]