Short Communications
Scand. J. Dent. Res. 1978: 86: 412-414 (Kcv wortis: dentifrice; Streptococcus mutans; toothbrush)
Contamination of toothpaste and toothbrush by Streptococcus mutans M. SVANBERG Department of Cariology, Faculty of Odontology, University of Gothenburg, Gothenburg, Sweden
ABSTRACT - Toothbrushes and toothpaste tubes used by persons infected with S. mutans were examined for the presence of this microorganism. Fifteen minutes after brushing > lO*" S. mutans were isolated from the toothbrushes and after ordinary storage for 24 h l C were recovered. From two out of 10 toothpaste tubes S. mutans was isolated from the orifice of tlie tube. The implications of these findings for the spread of the microorganism are discussed. (Accepted for publication 22 April 1978)
The toothbrush becomes heavily infected wilh microorganisms from the tooth surfaces (DAYOUB, RusiKO & GROSS 1977). According to APPLETON (1950) the toothbrush has been responsible for the transmission of infections like syphilis and diphtheria. Even though the use of the same toothbrush is mostly confined to one individual, the same toothpaste tube is generally used by several family members. As dental caries can be regarded as an infectious disease and S. mutans as a dental pathogen (KEYES 1960, ZiNNER, JABLON, ARAN & SASLAW 1965, KRASSE 1966) it seemed to be of interest to examine whether the toothpaste and the toothbrush can be contaminated with 5. mutans. Material and methods Two subjects with a salivary S. mutans concentration of 10^ colony forming units (CFU)
per ml were each given three new toothbrushes. After being used once in the routine oral hygiene procedure in the morning of three consecutive days the toothbrushes were kept dry at room temperature for 15 min, 12 and 24 h. Thereafter the heads of the tcjothbrushes were placed in 0.05 M phosphate buffer, vibrated in a Whirlimixer® and serially diluted in the phosphate buffer. Their ordinary toothpaste tubes were collected from 10 subjects with salivary S. mutans concentrations between 10^ and 10^ CFU per ml. The toothpastes were of seven different brands. Within 1 h after the tootlipastes had been used in the routine oral hygiene procedure of the subjects the opening of tlie tubes and the tops were streaked on Mitis Sahvarius Bacitracin agar with sucrose (MSB) (GOLD,JORDAN& HOUTE 1973).
All bacterial samples were cultured on MSB agar and incubated in anaerobic jars (596 COj+Nj) at 37°C for 48 h. The 5. mutans identification was performed according to the characteristic colony morphology on MSB agar. ImmunoBuorescent identification of S. mutans (BRATTHALL 1972) was performed
of
CONTAMINATION OF TOOTHBRUSH representative isolates using antisera provided by D. BRATTHALL.
Results
Two out of 10 toothpastes harbored .S. mutans. These toothpastes were used by the most heavily infected subjects with salivary S. mutans concentrations of about 10^ CFU per ml. No connection between the brand of toothpaste and the retention of 5. mulans was observed. Isolates from the toothpastes and the toothbrushes of the two subjects who had used the S. mutans positive toothpastes belonged to serotype c. On the toothbrushes S. mutans represented 1.5 and 6.0 X 10^ CFU 15 min after the cleaning of the teeth. The S. mutans CFU decreased with time to 0.8 and 5.0 X 10* 12 h after and to 0.9 and 4.0 X 10* 24 h after usage of the toothbrushes.
Discussion
The findings show that toothbrushes can be heavily infected with 5. mutans for a long time after usage and that the toothpaste also can harbor this microorgatiism. With existing knowledge about the spread of S. mutans these observations are of a certain interest. In the oral cavity S. mutans preferably colonizes tooth surfaces (CARLSSON 1967) and therefore becomes established at the time teeth erupt (BERKOWITZ, JORDAN &; WHITE
1975,
CARLSSON,
GRAHNEN
&
JONSSON 1975). Bacteriocin typing studies indicate the mother as a source of infection for her child (BERKOWITZ & JORDAN 1975). Salivary contaminated objects like metal spoons were found to
413
harbor S. mutans for several hours and were suggested as sources of infection between parent and child (KOEHLER &: BRATTHALL 1978). Interindividual transmission of S. mutans may, however, also occur via the contaminated toothpaste and toothbrush. On the tooth surfaces the toothbrush at the same time disturbs or reduces the existing flora and introduces other microorganisms like S. mutans. The disturbance of the existing flora may result in removal of competitive organisms and an increased possibility for S. mutans to colonize. It is difficult to artificially establish S. mutans (KRASSE, EDWARDSSON, SVENSSON & TRELL 1967,
EDMAN, KEENE, SHKLAIR&
HOERMAN 1975, SvANBERG 8c LOESCHE 1978a,b). KRASSE et al. (1967) successfully implanted the organism first during a regimen with frequent sucrose ingestions and after suppression of the existing flora by polishing of the teeth. EDMAN et al. (1975) implanted S. mutans into approximal areas by the use of dental floss, which may be another way to disturb the microbial ecology on the tooth surface and thereby favor the colonization of the new organisms on the tooth surface. The intraindividual spread of S. mutans does not occur easily either. The organism has been consistently recovered from certain tooth sites but not from others in the same mouth (GIBBONS, DEPAOLA, SPINELL & SKOBE 1974) and implanted S. mutans have spread only to adjacent tooth surfaces (EDMAN et al. 1975, SvANBERG Sc LOESCHE 1978a,b). The dental explorer has been shown to be an effective tool to spread 5. mutans from an infected to a noninfected site in the oral cavity (LOESCHE & SvANBERG 1977). EDMAN et al. (1975) suggested the possibility that improper use of dental floss may also help to spread this organism. If this is the case, and if the use of other ordinary oral
SVANBERG
414
hygiene methods might facilitate the spread of pathogenic microorganisms, this warrants further study.
& SKOBE, Z.: Interdental localizadon of Streptococcus mutans as related to dental carries experience. Infect. Immunol. 1974: 9: 481-488. GOLD, O . G., JORDAN, H . V. & HOUTE, J. VAN:
References APPLETON, J . L. T . :
Bacterial infection. Lea & Febiger, Philadelphia 1950, p. 625. BERKOWITZ, R.J. & JORDAN, H . V.: Similarity of bacteriocins of Streptococcus mutans from mother and infant. Arch. Oral Biol. 1975:20: 725-730. BERKOWITZ, R.J., JORDAN, H . V. & WHITE, G . :
The early establishment of Streptococcus mutans in the mouths of infants. Arch. Oral Biol. 1975: 20: 171-174. BRATTHALL,
D.:
Serological
studies
on
Streptococcus mutans. Odontol. Revy 1972: 23: Suppl. 23. CARLSSON, J.: Presence of various types of non-haemolytic streptococci in dental plaque and in other sites of the oral cavity in man. Odontol. Revy 1967: 18: 55-74. CARLSSON, J.,
GRAHNEN, H . & JONSSON, G. :
Lactobacilli and streptococci in the mouth of children. Caries Res. 1975: 9: 333-339. DAYOUB,
D.,
RUSIKO,
D. B. & GROSS,
A.:
Microbial contamination of toothbrushes. J. Dent. Res. \911: 5 6 : 706. EDMAN, D. C , KEENE, H . J . , SHKLAIR, I. L. &
HoERMAN, K. C : Dental floss for implantation and sampling of Streptococcus mutans from approximal surfaces of human tceih. Arch. Oral Biol. 1975:20: 145-148. GIBBONS, R. J., DEPAOLA, P. F., SPINELL, D . M .
Address: M. Svanberg Odontologiska Kliniken Fack 40033 Gothenburg 33 Sweden
A selective medium for Streptococcus mutans. Arch. Oral Biol. 1973: 18: 1357-1364. KEYES, P. H . : The infectious and transmissible nature of experimental dental caries: hndings and implications. Arch. Oral Biol. 1960: 1: 304-320. KRASSE, B.: Human streptococci and experimental caries in hamster. Arch. Oral Biol. 1966: 11: 429-436. KRASSE, B., EDWARDSSON, S., SVENSSON, I. & TRELL, L . : Implantation of caries inducing
streptococci in the human oral cavity. Arch. Oral Biol. 1967: 12: 213-236. KoEHLER, B. & BRATTHALL, D . : Intrafamilial
levels of Streptococcus mutans and some aspects of the bacterial transmission. Scand. J. Dent. Res. 1978: 86: 35-42. LOESCHE, W. J. &: SvANBERG, M.: Intraoral spread of Strep, mutans by means of a dental explorer./. Dent. Res. 1977: 56: Spec. Issue B. Abstr. No. 2. p. 2. SvANBERG, M. & LoESCHE, W. J.: Implantation of Strep, mutans in humans. Arch. Oral Biol. 1978a: in press. SvANBERG, M. & LoESCHE, W. J.: Intraoral spread of Strep, mutans in humans. Arch. Oral Biol. 1978b: in press. ZiNNER, D. D., JABLON, J. M., ARAN, A. P. &
S AS LAW, M. S.: Experimental caries induced in animals by streptococci of human origin. Proc. Soc. Exp.
766-770.
Biol.
Med.
1965: 118:
Scand. J. Dent. Res. 1978: 86: 412-414 (Kcv wortis: dentifrice; Streptococcus mutans; toothbrush)
Contamination of toothpaste and toothbrush by Streptococcus mutans M. SVANBERG Department of Cariology, Faculty of Odontology, University of Gothenburg, Gothenburg, Sweden
ABSTRACT - Toothbrushes and toothpaste tubes used by persons infected with S. mutans were examined for the presence of this microorganism. Fifteen minutes after brushing > lO*" S. mutans were isolated from the toothbrushes and after ordinary storage for 24 h l C were recovered. From two out of 10 toothpaste tubes S. mutans was isolated from the orifice of tlie tube. The implications of these findings for the spread of the microorganism are discussed. (Accepted for publication 22 April 1978)
The toothbrush becomes heavily infected wilh microorganisms from the tooth surfaces (DAYOUB, RusiKO & GROSS 1977). According to APPLETON (1950) the toothbrush has been responsible for the transmission of infections like syphilis and diphtheria. Even though the use of the same toothbrush is mostly confined to one individual, the same toothpaste tube is generally used by several family members. As dental caries can be regarded as an infectious disease and S. mutans as a dental pathogen (KEYES 1960, ZiNNER, JABLON, ARAN & SASLAW 1965, KRASSE 1966) it seemed to be of interest to examine whether the toothpaste and the toothbrush can be contaminated with 5. mutans. Material and methods Two subjects with a salivary S. mutans concentration of 10^ colony forming units (CFU)
per ml were each given three new toothbrushes. After being used once in the routine oral hygiene procedure in the morning of three consecutive days the toothbrushes were kept dry at room temperature for 15 min, 12 and 24 h. Thereafter the heads of the tcjothbrushes were placed in 0.05 M phosphate buffer, vibrated in a Whirlimixer® and serially diluted in the phosphate buffer. Their ordinary toothpaste tubes were collected from 10 subjects with salivary S. mutans concentrations between 10^ and 10^ CFU per ml. The toothpastes were of seven different brands. Within 1 h after the tootlipastes had been used in the routine oral hygiene procedure of the subjects the opening of tlie tubes and the tops were streaked on Mitis Sahvarius Bacitracin agar with sucrose (MSB) (GOLD,JORDAN& HOUTE 1973).
All bacterial samples were cultured on MSB agar and incubated in anaerobic jars (596 COj+Nj) at 37°C for 48 h. The 5. mutans identification was performed according to the characteristic colony morphology on MSB agar. ImmunoBuorescent identification of S. mutans (BRATTHALL 1972) was performed
of
CONTAMINATION OF TOOTHBRUSH representative isolates using antisera provided by D. BRATTHALL.
Results
Two out of 10 toothpastes harbored .S. mutans. These toothpastes were used by the most heavily infected subjects with salivary S. mutans concentrations of about 10^ CFU per ml. No connection between the brand of toothpaste and the retention of 5. mulans was observed. Isolates from the toothpastes and the toothbrushes of the two subjects who had used the S. mutans positive toothpastes belonged to serotype c. On the toothbrushes S. mutans represented 1.5 and 6.0 X 10^ CFU 15 min after the cleaning of the teeth. The S. mutans CFU decreased with time to 0.8 and 5.0 X 10* 12 h after and to 0.9 and 4.0 X 10* 24 h after usage of the toothbrushes.
Discussion
The findings show that toothbrushes can be heavily infected with 5. mutans for a long time after usage and that the toothpaste also can harbor this microorgatiism. With existing knowledge about the spread of S. mutans these observations are of a certain interest. In the oral cavity S. mutans preferably colonizes tooth surfaces (CARLSSON 1967) and therefore becomes established at the time teeth erupt (BERKOWITZ, JORDAN &; WHITE
1975,
CARLSSON,
GRAHNEN
&
JONSSON 1975). Bacteriocin typing studies indicate the mother as a source of infection for her child (BERKOWITZ & JORDAN 1975). Salivary contaminated objects like metal spoons were found to
413
harbor S. mutans for several hours and were suggested as sources of infection between parent and child (KOEHLER &: BRATTHALL 1978). Interindividual transmission of S. mutans may, however, also occur via the contaminated toothpaste and toothbrush. On the tooth surfaces the toothbrush at the same time disturbs or reduces the existing flora and introduces other microorganisms like S. mutans. The disturbance of the existing flora may result in removal of competitive organisms and an increased possibility for S. mutans to colonize. It is difficult to artificially establish S. mutans (KRASSE, EDWARDSSON, SVENSSON & TRELL 1967,
EDMAN, KEENE, SHKLAIR&
HOERMAN 1975, SvANBERG 8c LOESCHE 1978a,b). KRASSE et al. (1967) successfully implanted the organism first during a regimen with frequent sucrose ingestions and after suppression of the existing flora by polishing of the teeth. EDMAN et al. (1975) implanted S. mutans into approximal areas by the use of dental floss, which may be another way to disturb the microbial ecology on the tooth surface and thereby favor the colonization of the new organisms on the tooth surface. The intraindividual spread of S. mutans does not occur easily either. The organism has been consistently recovered from certain tooth sites but not from others in the same mouth (GIBBONS, DEPAOLA, SPINELL & SKOBE 1974) and implanted S. mutans have spread only to adjacent tooth surfaces (EDMAN et al. 1975, SvANBERG Sc LOESCHE 1978a,b). The dental explorer has been shown to be an effective tool to spread 5. mutans from an infected to a noninfected site in the oral cavity (LOESCHE & SvANBERG 1977). EDMAN et al. (1975) suggested the possibility that improper use of dental floss may also help to spread this organism. If this is the case, and if the use of other ordinary oral
SVANBERG
414
hygiene methods might facilitate the spread of pathogenic microorganisms, this warrants further study.
& SKOBE, Z.: Interdental localizadon of Streptococcus mutans as related to dental carries experience. Infect. Immunol. 1974: 9: 481-488. GOLD, O . G., JORDAN, H . V. & HOUTE, J. VAN:
References APPLETON, J . L. T . :
Bacterial infection. Lea & Febiger, Philadelphia 1950, p. 625. BERKOWITZ, R.J. & JORDAN, H . V.: Similarity of bacteriocins of Streptococcus mutans from mother and infant. Arch. Oral Biol. 1975:20: 725-730. BERKOWITZ, R.J., JORDAN, H . V. & WHITE, G . :
The early establishment of Streptococcus mutans in the mouths of infants. Arch. Oral Biol. 1975: 20: 171-174. BRATTHALL,
D.:
Serological
studies
on
Streptococcus mutans. Odontol. Revy 1972: 23: Suppl. 23. CARLSSON, J.: Presence of various types of non-haemolytic streptococci in dental plaque and in other sites of the oral cavity in man. Odontol. Revy 1967: 18: 55-74. CARLSSON, J.,
GRAHNEN, H . & JONSSON, G. :
Lactobacilli and streptococci in the mouth of children. Caries Res. 1975: 9: 333-339. DAYOUB,
D.,
RUSIKO,
D. B. & GROSS,
A.:
Microbial contamination of toothbrushes. J. Dent. Res. \911: 5 6 : 706. EDMAN, D. C , KEENE, H . J . , SHKLAIR, I. L. &
HoERMAN, K. C : Dental floss for implantation and sampling of Streptococcus mutans from approximal surfaces of human tceih. Arch. Oral Biol. 1975:20: 145-148. GIBBONS, R. J., DEPAOLA, P. F., SPINELL, D . M .
Address: M. Svanberg Odontologiska Kliniken Fack 40033 Gothenburg 33 Sweden
A selective medium for Streptococcus mutans. Arch. Oral Biol. 1973: 18: 1357-1364. KEYES, P. H . : The infectious and transmissible nature of experimental dental caries: hndings and implications. Arch. Oral Biol. 1960: 1: 304-320. KRASSE, B.: Human streptococci and experimental caries in hamster. Arch. Oral Biol. 1966: 11: 429-436. KRASSE, B., EDWARDSSON, S., SVENSSON, I. & TRELL, L . : Implantation of caries inducing
streptococci in the human oral cavity. Arch. Oral Biol. 1967: 12: 213-236. KoEHLER, B. & BRATTHALL, D . : Intrafamilial
levels of Streptococcus mutans and some aspects of the bacterial transmission. Scand. J. Dent. Res. 1978: 86: 35-42. LOESCHE, W. J. &: SvANBERG, M.: Intraoral spread of Strep, mutans by means of a dental explorer./. Dent. Res. 1977: 56: Spec. Issue B. Abstr. No. 2. p. 2. SvANBERG, M. & LoESCHE, W. J.: Implantation of Strep, mutans in humans. Arch. Oral Biol. 1978a: in press. SvANBERG, M. & LoESCHE, W. J.: Intraoral spread of Strep, mutans in humans. Arch. Oral Biol. 1978b: in press. ZiNNER, D. D., JABLON, J. M., ARAN, A. P. &
S AS LAW, M. S.: Experimental caries induced in animals by streptococci of human origin. Proc. Soc. Exp.
766-770.
Biol.
Med.
1965: 118:
