ISSN 2413-6077. IJMMR 2017 Vol. 3 Issue 150

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dOI 10.11603/IJMMR.2413-6077.2017.1.6955

INfLUENcE Of LactobaciLLus sPP. ON cOLONIZATION  
ANd ANTI-INfEcTIOUs REsIsTANcE Of ThE mUcOUs mEmBRANEs  
Of ThE UPPER REsPIRATORY TRAcT

S. V. Kalinichenko1, O. O. Korotkykh1, S. I. Pokhil1, M. G. Bakumenko2
1NATIONAL ACADEMY OF MEDICAL SCIENCES OF UKRAINE, 

1MECHNIKOV INSTITUTE OF MICROBIOLOGY AND IMMUNOLOGY, KHARKIV, UKRAINE
2KHARKIV CITY STUDENT HOSPITAL, KHARKIV, UKRAINE

Background. Lactobacilli are very important for the formation of colonization resistance and have pro-
nounced antagonistic effect against a wide range of microorganisms. That is why the lactobacilli have extensive 
use as a component of classic probiotic agents that are widely used to prevent and treat dysbiotic conditions of 
digestive and genital systems of people. 

Objective. The aim of the research was to study the effect of lactobacilli on anti-infectious resistance of 
mucous membranes of upper respiratory tract.

Methods. The colonization degree (lg CFU / g) of nasal mucosal membranes by Lactobacillus spp. and S. 
aureus was determined in all carriers before the experiment. Also, the level of lysozyme and secretory immuno-
globulin A (sIgA) in nasal secretions cavities was identified. 

Results. It was established a clear dysfunction of anti-infectious resistance in carriers of Staphylococcus 
aureus - a decrease of colonization resistance and local immunity of mucous membranes of upper respiratory 
tract. As for the anti-infectious resistance of nasal mucosal of S. aureus carriers, the level of lysozyme and secre-
tory immunoglobulin A gradually increased after the application of probiotic strain L. rhamnosus GG, and in 21 
days it reached rates of healthy individuals. 

Conclusions. It was found out that probiotics for nasal passages sanitation in Staphylococcus aureus car-
riers lead to gradual eradication of the pathogen (S. aureus) with restoration of colonization and anti-infectious 
resistance, mucous membranes and upper respiratory tract.

KEYWORDS: Lactobacillus spp.; lysozyme; sIgA; upper respiratory tract.

Corresponding author: Svitlana Kalinichenko, Laboratory of 
Viral Infections, National Academy of Medical Sciences of 
Ukraine Mechnikov Institute of Microbiology and Immunology, 
14 Pushkinska Street, Kharkiv, Ukraine, 61057.
Phone number: +380577313151
E-mail: imidir@ukr.net

Introduction
Lactobacilli are widely distributed in the 

environment and have a high biological activ-
ity. They are an important part of normal mi-
croflora of digestive and genital human tracts 
and they belong to the resident microflora of 
nasopharynx [1]. Lactobacilli are actively in-
volved in the formation of colonization resist-
ance of mucous membranes and have signifi-
cant antagonistic action against a wide range 
of bacteria [2-4]. That is why the lactobacilli are 
widely used as a component of classic probi-
otic agents [3-5]. Probiotics based on lactoba-
cilli are widely used to prevent and treat dysbi-
otic conditions of digestive and genital human 

system [6-8]. Nowadays, research on the use of 
lactobacilli for prevention or treatment of infec-
tions and recovery of anti-infectious resistance 
of mucous membranes of upper respiratory 
tract (URT) are actual [9]. 

The aim of the research was to study the 
effect of Lactobacillus spp. on colonization and 
anti-infectious resistance of mucosal mem-
branes of upper respiratory tract in Staphylococ-
cus aureus carriers. The objects of the study 
were the carriers of Staphylococcus aureus 
(n=29) among the medical staff of a hospital, 
Kharkiv (Ukraine). 

Methods
The colonization degree (lg CFU / g) of nasal 

mucosal membranes by Lactobacillus spp. and 
S. aureus was determined in all the carriers 
before the experiment. Also, the level of 
lysozyme and secretory immunoglobulin A 

S. V. Kalinichenko et al.

International Journal of Medicine and Medical Research 
2017, Volume 3, Issue 1, p. 50–52
copyright © 2017, TSMU, All Rights Reserved



ISSN 2413-6077. IJMMR 2017 Vol. 3 Issue 1 51

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S. V. Kalinichenko et al.

(siga) in nasal secretions cavities was identified. 
then the carriers were divided into groups: i 
(n=7, control) – the persons, who received saline 
as the sanitation agent (0.85% NaCl, pH 7.2) 
during 30 days, 2–3 drops twice per day in each 
nostril; II (n=11) – the persons, who received as 
the sanitation agent a suspension of probiotic 
strain L. rhamnosus GG (in saline) at a concen-
tration of 5×109 CFU/ml (according to the group 
I scheme). The degree of colonization of nasal 
mucous membranes by Lactobacillus spp. and 
S. aureus, lysozyme and sIgA levels in nasal 
cavities secretions were determined in the in-
dividuals taking part in the experiments repeat-
edly in 7, 14 and 21 days of sanitation. The 
material was selected as follows: in each nostril, 
alternatively, with special replicator sterile foam 
tape was injected and mucus impregnation was 
held for 5 minutes, after it the tape was im-
mersed in a sterile tube containing 1.0 ml of 
phosphate-buffered saline (PBS, pH 7.2) for 30 
minutes and then separated from the foam 
carrier mucus by insulin syringe, forcing it into 
the PBS. 

In the comparisons group there were per-
sons, who did not have upper respiratory tract 
infection within 6 months (almost healthy, 
n=15).

Lysozyme levels were determined by means 
of nephelometric method [10]. The concentra-
tion of lysozyme was rated in micrograms/ml 
(μg/ml). to determine siga, RiDaSCREEn®siga 
ELISA kits of R-Biopharm AG Company (Ger-
many) were used. The results of calculations 
were carried out on Stat Fax enzyme immuno-
assay analyser. sIgA concentration was rated 
in mg/ml.

Results
It was established that in the healthy indivi-

duals (n=15) the degree of colonization of the 
na sal mucous membranes Lactobacillus spp. 
was 6.64±0.9 lg CFU/g, the amount of lysozyme 
and sIgA in nasal secretions cavities were res-
pec tively at 17.45±2.9 μg/ml and 129.3±21.7 mg/ml.

The degree of colonization of nasal mucosa 
Lactobacillus spp. was 2.2±0.7 lg CFU/g, the 
amount of lysozyme and sIgA in nasal secre-

tions cavities were respectively 7.45±2.1 μg/ml 
and 63.9±19.8 mg/ml in the S. aureus carriers. 

Discussion
Consequently, the degree of colonization 

of nasal mucosa Lactobacillus spp. in the 
carriers of Staphylococcus aureus was in 3.8–5.0 
(p<0.01) times lower compared to the healthy 
individuals. The amount of lysozyme and sIgA 
in nasal secretions cavities were also lower in 
comparison with the healthy individuals, res-
pec tively in 1.5–3.8 (p<0.05), and 1.2–3.4 (p<0.05) 
times.

Thus, it was established a clear dysfunction 
of anti-infectious resistance in the carriers of 
Staphylococcus aureus: the decrease of coloni-
zation resistance and local immunity of mucous 
membranes of upper respiratory tract.

According to the literature, application of 
probiotics or synbiotic treatment regimens in 
respiratory infections reduces relapse respi-
ratory tract infections and bronchial obstruction 
in children [9]. But the authors did not conduct 
any studies to determine the number of lac-
tobacilli in secrets of URt and their influence 
on anti-infectious resistance of mucous mem-
branes. In connection with the foregoing, we 
conducted a study on the possibility of recovery 
colonization and anti-infectious resistance of 
mucous membranes of upper respiratory tract 
in the carriers of S. aureus using the probiotic 
strain L. rhamnosus GG.

The data in the Table 1 prove that the pro-
biotic strain L. rhamnosus GG for sanitation the 
carriers decreased the degree of colonization 
by Staphylococcus aureus mucosal in 7 and 14 
days after the sanitation of lactobacilli suspen-
sion, and in 21 days there was a complete era-
dication of S. aureus from nasal mucous mem-
branes. Moreover, the decrease of S. aureus 
degree colonization on mucosal of the carriers 
occurred against the background of increase 
in the number of Lactobacillus spp. on mucous 
membranes.

As for the anti-infectious resistance of nasal 
mucosal of S. aureus carriers, the level of lyso-
zyme and secretory immunoglobulin A after 
the application of the probiotic strain L. rham-

Table 1. Average indexes of mucosal colonization resistance after sanitation, (M±m)

Groups
The degree of mucosal colonization URT, lg CFU / g

Lactobacillus spp. S. aureus
in 7 days in 14 days in 21 days in 7 days in 14 days in 21 days

I 2.3±0.7 2.2±0.6 2.1±0.7 4.7±0.7 4.9±0.6 4.8±0.6
II 3.8±0.4* 4.5±0.4* 5.3±0.4* 1.8±0.6* 0.6±0.6* 0±0*

note: * – significant difference between these indexes of i and ii groups (p<0.05).



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nosus gg significantly increased, and in 21 days 
it reached rates of healthy individuals (Table 2).

To be precise, after the sanitation of probi-
otic nasal of the carriers Staphylococcus aureus, 
the level of lysozyme and siga rose: 7 days, an 
average of 1.6 (p <0.05) and 1.4 (p<0.05) times, 
respectively; at 14 days of 1.8 (p<0.05) and 1.6 
(p<0.05) times, respectively; after 21 days – 2.1 
(p<0.05) and 1.8 (p<0.05) times, respectively, 
compared with the group the carriers, that 
sanitation was conducted with saline.

Conclusions
 Thus, it was found that the application of 

probiotics for nasal passages sanitation in the 

Table 2. Average indexes of mucosal anti-infectious resistance after sanitation, (M±m)

Group
Indicator of local immunity

level of lysozyme, μg/ml level of sIgA, mg/ml
in 7 days in 14 days in 21 days in 7 days in 14 days in 21 days

I 7.3±2.4 7.4±2.2 7.4±2.3 61.3±21.3 62.1±18.8 61.9±19.3 
II 11.6±3.3* 13.3±2.9* 15.4±2.7* 84.8±19.8* 102.9±21.1* 112.2±19.1*

note: * – significant difference between these indexes of i and ii groups (p<0.05).

carriers of S. aureus leads to the gradual erad-
ication of the pathogen (S. aureus) with coloni-
zation restoration (increased degree of mucosal 
lactobacilli settlement), and anti-infectious re-
sistance (increased level of sIgA and lysozyme) 
in mucous membranes and upper respiratory 
tract.

This study demonstrated the positive effect 
of Lactobacillus spp. on the formation of 
colonization and anti-infectious resistance of 
upper respiratory tract mucous membranes 
and provided an opportunity to consider 
probiotics as a useful option for immune 
exposure to chronic infections of staphylococcal 
origin.

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Received: 2016-11-18

S. V. Kalinichenko et al.