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© Borgis - Nowa Stomatologia 3/2017, s. 120-128
*Elżbieta Pels, Izabela Tkacz-Ciebiera
Early childhood caries vs. the number of colonies of Streptococcus mutans and Lactobacillus spp. in saliva
Próchnica wczesnego dzieciństwa w odniesieniu do liczebności bakterii Streptococcus mutans i Lactobacillus spp. w ślinie
Department of Paedodontics, Medical University of Lublin
Head of Department: Professor Maria Mielnik-Błaszczak, MD, PhD
Wstęp. Próchnica wczesnego dzieciństwa (ECC) jest obecnym na całym świecie, szeroko rozpowszechnionym problemem zdrowotnym.
Cel pracy. To badanie stawia hipotezę, że rozwój ECC jest ściśle związany z rozwojem bakterii próchnicotwórczych Streptococcus mutans i Lactobacillus spp.
Materiał i metody. W badaniu uczestniczyło 346 dzieci w wieku od 7. do 48. miesiąca życia. Na podstawie danych zebranych w badaniu klinicznym oceniano frekwencję próchnicy oraz intensywność próchnicy zębów mlecznych. Uzyskane dane zostały poddane analizie statystycznej w odniesieniu do liczebności kolonii bakteryjnych Streptococcus mutans i Lactobacillus spp.
Wyniki. Badanie potwierdziło istotny statystycznie wzrost czynnych ognisk próchnicowych zębów mlecznych (liczba p) przy wzroście liczby kolonii bakteryjnych Streptococcus mutans oraz Lactobacillus spp. w ślinie badanych dzieci. Analiza statystyczna wykazała, że liczebność bakterii Lactobacillus spp. w ślinie zwiększa ryzyko nasilenia ECC średnio o 57,017 raza, natomiast obecność Streptococcus mutans – 209-krotnie.
Wnioski. Zwiększona intensywność próchnicy wczesnego dzieciństwa była istotnie powiązana ze zwiększoną liczebnością bakterii Streptococcus mutans i Lactobacillus spp. w ślinie dzieci w wieku żłobkowym. Należy zintensyfikować działania edukacyjne skierowane do rodziców małych dzieci dotyczące profilaktyki i leczenia choroby próchnicowej, aby zwiększyć szanse na zachowanie zdrowego uzębienia mlecznego do okresu fizjologicznej wymiany.
Introduction. Early childhood caries (ECC) is a widespread health problem all over the world.
Aim. The present study proposes a hypothesis that ECC is closely related to cariogenic bacteria Streptococcus mutans and Lactobacillus spp.
Material and methods. The investigation was carried out in a group of 346 children, aged 7-48 months. Based on the data collected in clinical examination, the prevalence of caries in deciduous teeth and its intensity were evaluated. Statistical analysis was carried out to assess the number of bacterial colonies of Streptococcus mutans and Lactobacillus spp.
Results. The investigation confirmed a statistically significant increase in the number of active foci of decay in deciduous teeth (d) at an increased number of bacterial colonies of Streptococcus mutans and Lactobacillus spp. detected in the saliva of the examined children. The statistical analysis revealed that the salivary content of Lactobacillus spp. increased the risk of ECC intensity 57.017 times, and Streptococcus mutans increased the risk 209 times.
Conclusions. Higher intensity of caries in early childhood is significantly associated with an increased count of cariogenic bacteria in the saliva of infants and children attending nursery schools. Therefore, more intensive education of parents and carers of young children is needed in terms of the prophylaxis and treatment of caries to facilitate preservation of healthy deciduous teeth until their physiological replacement by permanent dentition.

The development of dentition is long, complex, and closely related to the development and function of other organs involved in mastication. It depends upon various genetic, paragenetic and environmental factors (1, 2). Dental caries is a widespread health problem on a global scale. Some researchers believe dental caries to be the most widespread chronic disease among children (3). Early childhood caries (ECC) was first defined by the American Dental Association (ADA) and American Academy of Pediatric Dentistry (AADP) as the presence of one or more deciduous teeth affected by caries or resultant fillings, or a tooth that was extracted because of carious lesions in children aged 71 months or younger (2-8).
The fastest development of caries is favored by a combination of factors, including the presence of a great number of pathogenic bacteria, frequent exposition to sugar consumption and stagnation in the oral cavity due to improper oral hygiene habits, and weaker structure of dental tissues (7). Cariogenic bacteria are extremely potent, as they quickly transform sugars into acids (cariogenicity), are able to survive at a lower pH level (acidophilia), produce extracellular polysaccharides (EPS) and intracellular polysaccharides (IPS) (9). Streptococcus mutans and Streptococcus sobrinus are thought to be responsible for the initiation of dental decay. The infectivity window, i.e. the time of increased susceptibility of the oral cavity to colonization by cariogenic streptococcal species occurs between 7 months and 2 years of age (it is called the first window of infectivity). Another period of increased development of streptococcal colonies is observed between 6 and 12 years of age, when the first permanent teeth begin to erupt (the second window of infectivity) (10, 11). Nonetheless, in later studies, Milgrom et al. showed the colonization with S. mutans to occur in 25% of children even before the eruption of teeth (12).
Cross-sectional studies have shown a combination of factors to have a very complex effect on the development of caries, including the socioeconomic status, ethnic background, immigrant status, infant feeding patterns, exposure to fluoride, poor or inadequate oral hygiene, and the presence of EEC in children of preschool age (12-14).
The risk of dental caries is evaluated with various tools, e.g. microbiological examination, the rate of saliva secretion, pH of saliva, the condition of teeth and oral hygiene. Microbiological tests are performed to detect primarily bacterial species of Streptococcus mutans and Lactobacillus spp. Quantitative scores help to evaluate the degree of risk, and select those at the highest risk for dental caries. Moreover, a quantitative test measuring the number of Lactobacillus spp. colonies can be used to evaluate the diet, since the count of bacteria increases with higher consumption of carbohydrates, and persists for up to two weeks (4).
The study was aimed at the evaluation of early childhood caries prevalence and analysis of correlations between decayed deciduous teeth and the number of Streptococcus mutans and Lactobacillus spp. colonies in nursery school children.
Material and methods
The study was carried out in a group of 346 nursery school children, aged 7-48 months, mean age 26.28 ± 8.49 months. The examined group consisted of infants and children attending nurseries, 54.34% boys (n = 188), and 45.66% girls (n = 158). The deciduous teeth were clinically examined, with saliva samples collected and microbiologically examined.
The clinical examination of the current condition of the teeth was carried out by means of disposable diagnostic sets in artificial light. The findings were charted on examination sheets, and the following parameters were marked: deciduous teeth present or missing, active foci of caries, the number of teeth filled, and the number of teeth extracted due to caries. The prevalence of caries and intensity of decayed deciduous teeth was evaluated on the basis of the dmft index and its components: dt, mt, and ft.
Microbiological tests were carried out to evaluate the number of bacterial colonies of Streptococcus mutans in the saliva by means of Dentocult SM Strip mutans microbiological kit (Orion Diagnostica), and of Lactobacillus – with Dentocult LB (Orion Diagnostica). The samples were cultured, and the results read according to the instructions provided by the manufacturer. Considering the age of the subjects, the recommended method of saliva collection was modified. Paraffin chewing cubes were not used. Instead, the submandibular area and the area around the ears were massaged to obtain mixed saliva. The saliva was collected from the floor of the mouth into disposable syringes. The samples were cultured on a medium specific for S. mutans and Lactobacillus spp., and grown in an incubator at 37°C. SM tests were retrieved after 48 h, and LB tests after 4 days, and the density of colony forming units (CFU) per 1 ml of saliva was compared with the guide included in the testing kit.
Statistical analysis
The results of the clinical examination and microbiological scores were coded and analyzed statistically. To evaluate the differences between the analyzed groups for nominal features χ2 test was applied, and to establish the differences between two analyzed groups for quantitative features t-Student test was used. To determine the difference between several groups, one-way analysis of variance ANOVA was used. To establish the correlations between the parameters, Spearman’s test (R) was used.
To present dependences between multiple properties and demonstrate correlations between data, the odds ratio (OR) was calculated, with 5% conclusion error and p < 0.05 assumed as the level of significance. The database and statistical analyses were performed with Statistica 10.0 software (StatSoft, Poland).
All patients and their parents consented to the clinical examination and to the collection of saliva, in accordance with the Declaration of Helsinki. The study was approved by the local Bioethics Committee KE-0254/7/2014 at the Medical University of Lublin. The study was financed by Statutory Activities No. DS 291, Medical University of Lublin, Poland.
Clinical examination

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otrzymano: 2017-07-14
zaakceptowano do druku: 2017-08-04

Adres do korespondencji:
*Elżbieta Pels
Zakład Stomatologii Wieku Rozwojowego Uniwersytet Medyczny w Lublinie
ul. Karmelicka 7, 20-081 Lublin
tel. +48 (81) 532-06-19

Nowa Stomatologia 3/2017
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