Galioto GB (ed): Tonsils: A Clinically Oriented Update. Adv Otorhinolaryngol. Basel, Karger, 1992, vol 47, pp 168-171
Alpha-Streptococci -Inhibiting Beta-Streptococci Group A in Treatment of Recurrent Streptococcal Tonsillitis Helena Lilja a, Eva Grahn a, Stig E. Holm a, Kristian RooSb a Department
of Clinical Microbiology, University of Umea, and bENT Department, Lundby Hospital, Goteborg, Sweden
The recurrence rate after penicillin treatment of streptococcal tonsillitis has been reported to be 10-25% [1] and several factors have been proposed to contribute to this [2]. One important factor is ecological disturbances following antibiotic treatment. The normal bacterial flora is an important barrier towards invading beta-hemolytic streptococci group A (GAS). We demonstrated earlier in vitro that alpha-streptococci can inhibit GAS in vitro [3] and that the absence of interfering alpha-streptococci correlated to treatment failure in streptococcal tonsillitis [4]. Therefore we have isolated from throat samples obtained from healthy individuals four alpha-streptococci with a strong growth-inhibiting capacity against clinical isolates of GAS. The aim of this study was to see if the isolated alpha-streptococci inhibit GAS independent of their serotype and geographic area of recovery. Material and Methods
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Bacterial Strains Four alpha-streptococci with broad growth-inhibiting capacity versus GAS [3] were used in the present study. These represented three Streptococcus sanguis II and one Streptococcus mitis according to the API Strep Test (France). 392 GAS strains were isolated from patients with tonsillitis from four different cities in Sweden: A-D (A = Uppsala, B = Umea, C = Goteborg, D = Boden). The GAS strains were T-typed using T-typing sera (Institute of Sera and Vaccines, Prague, Czechoslovakia) and SOR-tested, all according to Maxted and Widdowson [5].
Alpha-Streptococci in Streptococcal Tonsillitis
% 100
,.....-
-
.--
169
r--
75
50
25
o
A
B
City
C
D
Fig. 1. Total inhibition of 392 GAS strains by four alpha-streptocci. A = Uppsala, B= Umea, C = Goteborg, D = Boden.
Interference GAS and alpha-streptococci were grown in a trypticase-yeast medium at 37°C. Minidrops of the alpha-streptococcal strains (containing 10 5 CFU/ml) were transferred to blood agar plates (25 strains on each plate) by a Steers' steel pin replicator and allowed to dry. A sample of a GAS strain was applied adjacent to each of the alpha-streptococci and the plates were incubated in 5 % CO 2 at 37 °C. After one night's incubation the plates were examined for interference between the microorganisms. Inltibition of GAS growth was recorded as positive (+) and noninhibition of GAS or inhibition of the alpha-streptococci were recorded as negative (-). The combined GAS inhibiting capacity of all four alphastreptococci was designated 'total inhibition.'
Twenty-one different serotypes of GAS were found among the 392 isolates. Among those types, TlMl, Tl2Ml2 and T28 were the most frequent ones. The growth inhibition afforded by the four alpha-streptococci together (total inhibition) varied between 84 and 98 % of the GAS isolates from the four geographic areas (fig. l). The inhibitory capacity of alpha 2 and 3 was high against all GAS isolates but varied for alpha 4 significantly with the geographic area from which the GAS were recovered (fig. 2). All four alpha-streptococci had the capacity to inhibit the growth of GAS independent of serotype but the inhibition was less pronounced against the TlMl isolates (fig. 3).
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Results
LiljaiGrahniHolm/Roos
170
100
% 100
100 100 100
100
100
100
0
100
100
100 City
2
100
.1
[]2
1
~3
%
04
75
50
25
o
GAS
capacity
capacity
Fig. 2. Inhibiting capacity on GAS afforded by the four individual alpha-streptococci (1-4). A = Uppsala, B = Umea, C = Goteborg, D = Boden. Fig. 3. Inhibiting capacity of the four alpha-streptococci on various GAS serotypes.
Downloaded by: Université de Paris 193.51.85.197 - 1/16/2020 10:08:17 AM
3
Alpha-Streptococci in Streptococcal Tonsillitis
171
Conclusions
Although the four alpha-streptococci individually inhibited the growth of GAS to a various extent (11-98 %) they together had the capacity to inhibit 84-98 % of the 392 clinical isolates recovered from various geographic areas in Sweden. In a preliminary study these four alpha-streptococci have been successfully used for colonization to hinder further recurrences of streptococcal tonsillitis [6]. The present study underlines the importance of using more than one inhibiting alpha-streptococcal strain in such recolonization studies in order to account for differences in growthinhibiting capacity afforded by individual alpha strains.
References
2 3
4
5
6
Kaplan EL, Gastanaduy AS, Huwe BB: The role of the carrier in treatment failures after antibiotic therapy of group A streptococci in the upper respiratory tract. J Lab Clin Med 1981 ;98:326-335. Holm SE, Grahn E: Recurrence of streptococcal tonsillitis after penicillin treatment - a multifacetted problem. Res Clin Forums 1991;12:37-48. Grahn E, Holm SE, Ekedahl C, Roos K: Interference of a-hemolytic streptococci isolated from tonsillar surface on ~-hemolytic streptococci. A methodological study. Zentralbl Bakteriol Hyg I [A] 1983;254:459-468. Roos K, Grahn E, Holm SE: Evaluation of beta-Iactamase activity and microbial interference in treatment failures of acute streptococcal tonsillitis. Scand J Infect Dis 1986;18:313-319. Maxted WR, Widdowson JP: The protein antigens of group A streptococci; in Wannamaker LW, Matsen JM (eds): Streptococci and Streptococcal Diseases. New York, Academic Press, 1972, pp 252-264. Roos K, Grahn E, Holm SE, Johansson H, Lind L: Interfering alpha-streptococci as a protection against recurrent streptococcal tonsillitis in children. Int J Pediatr Otorhinolaryngol 1992, in press.
Prof. Stig E. Holm, Department of Clinical Bacteriology, University of Umea, S-901 87 Umea (Sweden)
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2
Alpha-Streptococci -Inhibiting Beta-Streptococci Group A in Treatment of Recurrent Streptococcal Tonsillitis Helena Lilja a, Eva Grahn a, Stig E. Holm a, Kristian RooSb a Department
of Clinical Microbiology, University of Umea, and bENT Department, Lundby Hospital, Goteborg, Sweden
The recurrence rate after penicillin treatment of streptococcal tonsillitis has been reported to be 10-25% [1] and several factors have been proposed to contribute to this [2]. One important factor is ecological disturbances following antibiotic treatment. The normal bacterial flora is an important barrier towards invading beta-hemolytic streptococci group A (GAS). We demonstrated earlier in vitro that alpha-streptococci can inhibit GAS in vitro [3] and that the absence of interfering alpha-streptococci correlated to treatment failure in streptococcal tonsillitis [4]. Therefore we have isolated from throat samples obtained from healthy individuals four alpha-streptococci with a strong growth-inhibiting capacity against clinical isolates of GAS. The aim of this study was to see if the isolated alpha-streptococci inhibit GAS independent of their serotype and geographic area of recovery. Material and Methods
Downloaded by: Université de Paris 193.51.85.197 - 1/16/2020 10:08:17 AM
Bacterial Strains Four alpha-streptococci with broad growth-inhibiting capacity versus GAS [3] were used in the present study. These represented three Streptococcus sanguis II and one Streptococcus mitis according to the API Strep Test (France). 392 GAS strains were isolated from patients with tonsillitis from four different cities in Sweden: A-D (A = Uppsala, B = Umea, C = Goteborg, D = Boden). The GAS strains were T-typed using T-typing sera (Institute of Sera and Vaccines, Prague, Czechoslovakia) and SOR-tested, all according to Maxted and Widdowson [5].
Alpha-Streptococci in Streptococcal Tonsillitis
% 100
,.....-
-
.--
169
r--
75
50
25
o
A
B
City
C
D
Fig. 1. Total inhibition of 392 GAS strains by four alpha-streptocci. A = Uppsala, B= Umea, C = Goteborg, D = Boden.
Interference GAS and alpha-streptococci were grown in a trypticase-yeast medium at 37°C. Minidrops of the alpha-streptococcal strains (containing 10 5 CFU/ml) were transferred to blood agar plates (25 strains on each plate) by a Steers' steel pin replicator and allowed to dry. A sample of a GAS strain was applied adjacent to each of the alpha-streptococci and the plates were incubated in 5 % CO 2 at 37 °C. After one night's incubation the plates were examined for interference between the microorganisms. Inltibition of GAS growth was recorded as positive (+) and noninhibition of GAS or inhibition of the alpha-streptococci were recorded as negative (-). The combined GAS inhibiting capacity of all four alphastreptococci was designated 'total inhibition.'
Twenty-one different serotypes of GAS were found among the 392 isolates. Among those types, TlMl, Tl2Ml2 and T28 were the most frequent ones. The growth inhibition afforded by the four alpha-streptococci together (total inhibition) varied between 84 and 98 % of the GAS isolates from the four geographic areas (fig. l). The inhibitory capacity of alpha 2 and 3 was high against all GAS isolates but varied for alpha 4 significantly with the geographic area from which the GAS were recovered (fig. 2). All four alpha-streptococci had the capacity to inhibit the growth of GAS independent of serotype but the inhibition was less pronounced against the TlMl isolates (fig. 3).
Downloaded by: Université de Paris 193.51.85.197 - 1/16/2020 10:08:17 AM
Results
LiljaiGrahniHolm/Roos
170
100
% 100
100 100 100
100
100
100
0
100
100
100 City
2
100
.1
[]2
1
~3
%
04
75
50
25
o
GAS
capacity
capacity
Fig. 2. Inhibiting capacity on GAS afforded by the four individual alpha-streptococci (1-4). A = Uppsala, B = Umea, C = Goteborg, D = Boden. Fig. 3. Inhibiting capacity of the four alpha-streptococci on various GAS serotypes.
Downloaded by: Université de Paris 193.51.85.197 - 1/16/2020 10:08:17 AM
3
Alpha-Streptococci in Streptococcal Tonsillitis
171
Conclusions
Although the four alpha-streptococci individually inhibited the growth of GAS to a various extent (11-98 %) they together had the capacity to inhibit 84-98 % of the 392 clinical isolates recovered from various geographic areas in Sweden. In a preliminary study these four alpha-streptococci have been successfully used for colonization to hinder further recurrences of streptococcal tonsillitis [6]. The present study underlines the importance of using more than one inhibiting alpha-streptococcal strain in such recolonization studies in order to account for differences in growthinhibiting capacity afforded by individual alpha strains.
References
2 3
4
5
6
Kaplan EL, Gastanaduy AS, Huwe BB: The role of the carrier in treatment failures after antibiotic therapy of group A streptococci in the upper respiratory tract. J Lab Clin Med 1981 ;98:326-335. Holm SE, Grahn E: Recurrence of streptococcal tonsillitis after penicillin treatment - a multifacetted problem. Res Clin Forums 1991;12:37-48. Grahn E, Holm SE, Ekedahl C, Roos K: Interference of a-hemolytic streptococci isolated from tonsillar surface on ~-hemolytic streptococci. A methodological study. Zentralbl Bakteriol Hyg I [A] 1983;254:459-468. Roos K, Grahn E, Holm SE: Evaluation of beta-Iactamase activity and microbial interference in treatment failures of acute streptococcal tonsillitis. Scand J Infect Dis 1986;18:313-319. Maxted WR, Widdowson JP: The protein antigens of group A streptococci; in Wannamaker LW, Matsen JM (eds): Streptococci and Streptococcal Diseases. New York, Academic Press, 1972, pp 252-264. Roos K, Grahn E, Holm SE, Johansson H, Lind L: Interfering alpha-streptococci as a protection against recurrent streptococcal tonsillitis in children. Int J Pediatr Otorhinolaryngol 1992, in press.
Prof. Stig E. Holm, Department of Clinical Bacteriology, University of Umea, S-901 87 Umea (Sweden)
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2
