International Journal of Microbiology and Biotechnology

2020; 5(1): 16-23

http://www.sciencepublishinggroup.com/j/ijmb
doi: 10.11648/j.ijmb.20200501.13
ISSN: 2578-9678 (Print); ISSN: 2578-9686 (Online)

Antimicrobial Susceptibility Patterns of Alpha-Haemolytic

Streptococcus sp Isolated from Throat Swabs
Onwuka A. P.1 and Stanley C. N2
1
Corresponding Author Department of Medical Microbiology, University of Port Harcourt Teaching Hospital, Rivers state,
Nigeria. adaoraonwuka001@gmail.com
2
Department of Pharmaceutical Microbiology, Faculty of Pharmaceutical Sciences, University of Port Harcourt, Rivers State,
Nigeria

ABSTRACT
Alpha-hemolytic Streptococcus pneumoniae and a group of oral streptococci display alpha-hemolysis.
Their clinical importance includes acting as causative agents in associated clinical infections. The study
investigated the antibiotic susceptibility pattern of different antimicrobial agents against alpha-haemolytic
streptococci isolated from commensal flora of subjects presenting with tonsillitis at the University of Port
Harcourt Teaching Hospital. There was 28% growth of alpha-haemolytic Streptococcus sp among the 100
patients presenting with tonsillitis between January 2019-December 2019. The pattern of resistance
observed include 21% to Penicillin, 28.6% to Ofloxacin, 25.0% to Clindamycin, 17.9% to Ampicillin,
39.3% to Clarithromycin, 39.3% to Amoxyclav and Erythromycin, 53.6% to Cotrimoxazole and
Gentamycin, 39.2% to Vancomycin and 35.7% to Amikacin. The study showed considerable resistant
patterns of α-haemolytic Streptococcus sp associated with tonsillitis. This is a need to improve on
antimicrobial stewardship in our local communities across the country.
Keywords: Antibiotic resistance, Alpha-hemolytic streptococcus, Tonsillitis.

INTRODUCTION
Streptococci are a genus of Gram-positive bacteria that are widely distributed across the normal flora of
human and animals [1,2]. They cause diverse human diseases, and many of these species have the potential
to cause invasive infection resulting from the presence of bacteria in a normally sterile site [3,4]. Many
such invasive infections are of considerable public health importance. Alpha-hemolytic Streptococcus
pneumoniae and a group of oral streptococci (Streptococcus viridans or viridans streptococci) display
alpha-hemolysis [4-6]. Alpha-hemolysis is also termed incomplete hemolysis or partial hemolysis because
the cell membranes of the red blood cells are left intact. These organisms are generally classified as the
Viridians Group Streptococci (VGS) [7-9]. The VGS have been identified as mostly exhibiting alpha-
hemolysis on blood agar similar to Streptococcus pneumoniae, but having resistance to optochin and
insolubility in bile salts which are opposite characteristics of S. pneumoniae [9]. However, there exist
exceptions to these generalizations, as some VGS exhibit beta- or gamma-hemolysis and/or are sensitive to
optochin or bile [5,9].

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International Journal of Microbiology and Biotechnology
2020; 5(1): 16-21
http://www.sciencepublishinggroup.com/j/ijmb
doi: 10.11648/j.ijmb.20200501.13
ISSN: 2578-9678 (Print); ISSN: 2578-9686 (Online)

Viridans group streptococci comprise a vital part of the normal flora of the human upper respiratory tract
[3,5]. Their clinical importance includes acting as causative agents in infective endocarditis in patients with
both native and prosthetic valves and septicaemia in neutropenic patients [6,7]. In addition, they play a
significant role as a reservoir of antimicrobial resistance genes, transferring different resistance traits to
more pathogenic organisms like Streptococcus pneumoniae and Streptococcus pyogenes[7,8]. Tonsil
infections may be contagious, and can spread from person to person by contact with the mouth, throat, or
mucous of someone who is infected. Tonsillitis occur in up to 10% of the population frequently, while
reoccurrence could be as high as 50% [9,10]. Apart from treatment with antibiotics, tonsillectomy is one of
the most common inpatient surgery in childhood and adolescence under anaesthesia and the most common
service of ENT departments [11]. Resistance to penicillin and macrolides has been documented in viridans
streptococcal isolates from clinical specimens in different parts of the world [2–4,6]. Such resistance has
also been found in viridans streptococci from normal oral flora of children and adults with and without
infection [12,13]. In Finland, macrolide resistance has increased among S. pneumoniae and S. pyogenes
[14,15] and has also been found in group C and G β-haemolytic streptococci[16]. Three different resistance
mechanisms have been shown to cause macrolide resistance in these streptococcal isolates: target site
modification mediated by the erythromycin resistance methylases encoded by the erm(A) or erm(B) genes
conferring resistance to macrolide, lincosamide and streptogramin B antibiotics (MLSB phenotype); active
drug efflux mediated by the membrane-bound efflux protein encoded by the mef(A) gene conferring
resistance to 14- and 15-membered macrolides only (M phenotype); and mutations in the streptococcal 23S
rRNA or ribosomal protein genes leading to resistance to macrolide and streptogramin B antibiotics (MS
phenotype)[15–17]. In this study we investigated the antibiotic susceptibility pattern of different
antimicrobial agents against alpha-haemolytic streptococci isolated from commensal flora of subjects
presenting with tonsillitis at the University of Port Harcourt Teaching Hospital.
METHODS
Study Specimen
From January 2019 to December 2019, all cases showing growth of alpha-haemolytic Streptococcus sp
from throat swab samples was identified and reported for antimicrobial susceptibility at the University of
Port Harcourt Teaching Hospital was included in this study.
Antibiotic sensitivity
Antibiotic susceptibility pattern was determined by the disc diffusion method on Mueller-Hinton agar
according to the CLSI guidelines [18]. The antibiotics used were Penicillin (10µg -1), Ofloxacin (5µg-1),
Clindamycin (2µg-1), Ampicillin (10µg-1), Clarithromycin (15µg-1), Amoxyclav (30µg-1), Erythromycin
(15µg-1), Cotrimoxazole (25µg-1), Gentamycin (10µg-1), Vancomycin (30µg-1), Amikacin (30µg-1).

Data Analysis
The analysis was performed using SPSS software, version 20 (Chicago, IL, USA). The results are presented
using descriptive statistics in terms of relative frequency. Chi-square test was used to determine any
statistical association. Statistical significance was regarded as p values <0.05.

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International Journal of Microbiology and Biotechnology
2020; 5(1): 16-21
http://www.sciencepublishinggroup.com/j/ijmb
doi: 10.11648/j.ijmb.20200501.13
ISSN: 2578-9678 (Print); ISSN: 2578-9686 (Online)

RESULTS
Table 1: Demographic distribution of subjects

Frequency (n=100) Percentage (%)

Gender
Male 46 46.0
Female 54 54.0
Age-group
<1 year 24 24.0
1 - 16 years 72 72.0
17 - 25 years 2 2.0
> 25 years 2 2.0

Yes
No
28, 28.0%

72, 72.0%

Figure 1: Growth of α-haemolytic Streptococcus sp

Figure 1 shows there was 28% growth of alpha-haemolytic Streptococcus in the specimen collected
International Journal of Microbiology and Biotechnology
2020; 5(1): 16-21
http://www.sciencepublishinggroup.com/j/ijmb
doi: 10.11648/j.ijmb.20200501.13
ISSN: 2578-9678 (Print); ISSN: 2578-9686 (Online)

Table 2: Antibiotic sensitivity of Streptococcus sp isolated from throat swabs

Antibiotics Sensitive Intermediate Resistant

Penicillin (10) 20 (71.4) 2 (7.1) 6 (21.4)

Ofloxacin (5) 18 (64.3) 2 (7.1) 8 (28.6)

Clindamycin (2) 18 (64.3) 3 (10.7) 7 (25.0)

Ampicillin (10) 20 (71.4) 3 (10.7) 5 (17.9)

Clarithromycin (15) 14 (50.0) 3 (10.7) 11 (39.3)

Amoxyclav (30) 18 (64.3) 4 (14.3) 6 (21.4)

Erythromycin (15) 14 (50.0) 3 (10.7) 11 (39.3)

Cotrimoxazole (25) 4 (14.3) 9 (32.1) 15 (53.6)

Gentamycin (10) 4 (14.3) 9 (32.1) 15 (53.6)

Vancomycin (30) 14 (50.0) 3 (10.7) 11 (39.2)

Amikacin (30) 11 (39.3) 7 (25.0) 10 (35.7)

Chi-square = 533.6, p <0.0001; Difference is statistically significant (p<0.05)

DISCUSSION
Streptococcal infections are primarily treated based on the likely pathogens and their antibiotic
susceptibility, hence the vigilance of antimicrobial resistance patterns of alpha-haemolytic streptococcus to
various antibiotics is of the utmost importance [19]. The present comprehensive study provides a unique
opportunity to address the antibiotic susceptibility of α-haemolytic streptococcus isolated from different
subjects presenting with tonsillitis [20-22]. Antibiotic susceptibility testing of the isolates using the disc
diffusion method in the present study showed that 6 (21.4%) isolates was resistant to penicillin. In the
present study, the isolates were generally susceptible to antimicrobial agents except macrolides and
tetracycline; these data are similar to previous findings [21-23]. Worldwide data suggest an increased
resistance of GAS to macrolides, ranging from 0.5 to 70% [25-31]; a review of literature showed a gradual
increase up to 29.4% up until 2010 [26, 30, 31-34]. There was a 39.3% erythromycin resistance among the
isolates associated with tonsillitis among patients in the present study which is quite similar to previous
findings[27]. Moreover, the relatively high resistance to erythromycin might be a consequence of the
relatively frequent use of over the counter erythromycin. These therapy choices indicate that there is a mild
risk in prescribing erythromycin to patients. We observed a high percentage of isolates resistant to
Gentamycin, The high resistance (25.5 %) of GAS to tetracycline reported here is comparable to the global
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International Journal of Microbiology and Biotechnology
2020; 5(1): 16-21
http://www.sciencepublishinggroup.com/j/ijmb
doi: 10.11648/j.ijmb.20200501.13
ISSN: 2578-9678 (Print); ISSN: 2578-9686 (Online)

rise noticed [27,29,32] that has been assigned to polyclonal dissemination of resistance types, as antibiotic
resistance encoded by transposable elements does occur in α-haemolytic streptococcus [33]. The resistance
to vancomycin reported in this study are similar to results from previous reports[34]. We found that only
53.6% of GAS isolates were resistant to co-trimoxazole, as compared with 12.2% reported in Lucknow,
India[35]. Though resistance to co-trimoxazole was high, co-trimoxazole cannot be used because of the
intrinsic resistance in alpha-haemolytic streptococcus. Alpha hemolytic Streptococcus viridans (AHSV)
was found to be the predominant pathogen in neonatal septicemia and nosocomial infections in hospital-
based studies [32,35,36]. However, there have been a good correlation between institution of antibiotic
treatment and clinical improvement.
CONCLUSION
The study showed considerable resistant patterns of α-haemolytic Streptococcus sp associated with
tonsillitis. This is a need to improve on antimicrobial stewardship in our local communities across the
country.
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