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POPOVYCH, Andriy, RUZHYLO, Sofiya, ZAKALYAK, Nataliya, POPOVYCH, Dariya &
ŻUKOW, Xawery. Comparative study of the effects on the microbiota and leukocyturia of fresh and
canned Naftussya water. Quality in Sport. 2022;8(4):67-72. eISSN 2450-3118. DOI
https://dx.doi.org/10.12775/QS.2022.08.04.007
https://apcz.umk.pl/QS/article/view/41639

The journal has had 20 points in Ministry of Education and Science of Poland parametric evaluation. Annex to the announcement of the Minister of Education and Science of December 21, 2021. No. 32582.
Has a Journal's Unique Identifier: 201398. Scientific disciplines assigned: Economics and finance (Field of social sciences); Management and Quality Sciences (Field of social sciences).
Punkty Ministerialne z 2019 - aktualny rok 20 punktów. Załącznik do komunikatu Ministra Edukacji i Nauki z dnia 21 grudnia 2021 r. Lp. 32582. Posiada Unikatowy Identyfikator Czasopisma: 201398.
Przypisane dyscypliny naukowe: Ekonomia i finanse (Dziedzina nauk społecznych); Nauki o zarządzaniu i jakości (Dziedzina nauk społecznych).© The Authors 2022;
This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland

Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium,
provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike.
(http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited.

The authors declare that there is no conflict of interests regarding the publication of this paper.

Received: 15.11.2022. Revised: 20.12.2022. Accepted: 31.12.2022.

Comparative study of the effects on the microbiota and leukocyturia of fresh and
canned Naftussya water

Andriy I. Popovych1,2, Sofiya V. Ruzhylo3, Nataliya R. Zakalyak3, Dariya V. Popovych4,
Xawery Zukow5

1SE Ukrainian Scientific Research Institute of Medicine of Transport, Odesa, Ukraine
2Scientific group of Balneology of Hotel&Spa Complex “Karpaty”, Truskavets’, Ukraine

andriypopovych711@gmail.com
3Ivan Franko State Pedagogical University, Drohobych, Ukraine

doctor-0701@ukr.net natalyzak69@gmail.com
4IY Horbachevs’kyi National Medical University, Ternopil’, Ukraine

darakoz@yahoo.com
5Medical University of Bialystok, Bialystok, Poland xaweryzukow@gmail.com

Background. Balneotherapy has its rightful place in the arsenal of means of treatment of
chronic. The universally recognized gold standard of balneofactors is Naftussya bioactive
water. The issue of the loss of its healing properties by Naftussya water after extraction from
the subsoil, the possibility of its preservation and transportation remain debatable and relevant.
The purpose of this study is to compare the impact of fresh and canned water on the
microbiota of feces and urine in patients with chronic pyelonephritis. Materials and Methods.
The object of clinical-physiological observation were residents of the city of Truskavets’ (21
men aged 24-67 years and 8 women 33-76 years) with chronic pyelonephritis in remission.
The subject of the study were the leukocyturia and bacteriuria levels and components of
microbiota of feces. Results. Weekly use of fresh Naftussya water causes an increase in the
reduced content of probiotics in the microbiota and a decrease in the increased content of
conditionally pathogenic microflora, which is accompanied by a reduction of bacteriuria and
leveling of leukocyturia. The beneficial effect on the listed parameters of canned Naftussya
water is less pronounced, but the differences are statistically insignificant. Conclusion. The
obtained results provide grounds for continuing research into the effects of canned Naftusya
water on the body of urological patients.
Keywords: Naftussya bioactive water, chronic pyelonephritis, urinary syndrome, microbiota.

https://dx.doi.org/10.12775/QS.2022.08.04.007
https://apcz.umk.pl/QS/article/view/41639
http://creativecommons.org/licenses/by-nc-sa/4.0/)
mailto:andriypopovych711@gmail.com
mailto:doctor-0701@ukr.net
mailto:darakoz@yahoo.com
mailto:xaweryzukow@gmail.com


68

INRODUCTION

Balneotherapy has its rightful place in the arsenal of means of treatment of chronic
pyelonephritis [1]. The universally recognized gold standard of balneofactors is Naftussya
bioactive water. The issue of the loss of its healing properties by Naftussya water after
extraction from the subsoil, the possibility of its preservation and transportation remain
debatable and relevant [11,14,16-18]. The purpose of this study is to compare the impact of
fresh and canned water on the microbiota of feces and urine in patients with chronic
pyelonephritis.

MATERIALS AND METHODS

The object of clinical-physiological observation were residents of the city of Truskavets’
(21 men aged 24-67 years and 8 women 33-76 years) with chronic pyelonephritis in remission.
The day before, samples of morning urine and feces was collected, in which was determined
the leukocyturia and bacteriuria levels and components of microbiota respectively. Unified
methods are applied.

Urinary syndrome was assessed by quantitative and quantitative-qualitative [12] levels of
bacteriuria and leukocyturia. To qualitatively assess the manifestations of pyelonephritis, a
single-point IL Popovych’s [5,12] scale, built on the basis EC Harrington’s desirability
function [6], was used.

In particular, bacteriuria over 106 CFU/mL is quantified at 0,9 points (strongly expressed),
within (0,3÷1,0)•106 CFU/mL – 0,715 p (more than average, but not strong), 105 CFU/mL –
0,5 p (moderately expressed), (0,2÷0,5)•105 CFU/mL – 0,285 p (weakly expressed),
(0,01÷0,1)•105 CFU/mL - 0,1 p (very weak), less than 0,01•105 CFU/mL - 0 p (absent).
Leukocyturia over 60•103/mL - 0,715 р, within (20÷60)•103/mL – 0,5 р, (4÷20)•103/mL –
0,285 р, (2÷4)•103/mL – 0,1 р, less than 2•103/mL – 0 р.

The inclusion criteria were the presence of pronounced urinary syndrome (bacteriuria:
0,285÷0,715 points; leukocyturia: 0,1÷0,5 points) with preservation of functional renal
reserve (>10%), previously assessed by the AI Gozhenko method [4].

Every day, 6 patients were examined, who were divided into two groups, approximately
equal in terms of gender, age and severity of urinary syndrome. After the initial testing, the
members of the main group received a weekly course of balneotherapy [16] with fresh
Naftussya water (taken directly from the field), instead, for the experimental group, Naftussya
water, previously preserved in anaerobic conditions, was used. On the second day after the
end of the drinking course, repeated testing was carried out.

Normal (reference) values of variables are taken from the database of the Truskavetsian
Scientific School of Balneology.

For statistical analysis used the software package "Statistica 6.4".

RESULTS AND DISCUSSION

Both groups were equal in terms of gender (4 women each), age (M±SD: 52±15 and
50±10 years), body mass index (26,8±4,2 and 27,8±3,3 kg/m2) as well as of the initial
symptoms of urinary syndrome (Table 1) and components of the microbiota of feces (Table 2).



69

Table 1. Comparative characteristics of the symptoms of urinary syndrome

Naftussya water used for
balneotherapy

Student’s
Statistics

Reference
value

Symptons of
the urinary
syndrome

Fresh
(n=15)

Canned
(n=14)

t p Mean SD

Bacteriuria
actual,
lg CFU/mL

Before
After
Change

2,20±0,12***
1,10±0,24***
-1,10±0,22***

2,01±0,11***
1,22±0,25***
-0,79±0,20***

1,14
0,35
1,04

>0,2
>0,5
>0,5

0 0,98

Bacteriuria
qualitative,
points

Before
After
Change

0,51±0,05***
0,22±0,05***
-0,29±0,06***

0,48±0,04***
0,28±0,06***
-0,20±0,05***

0,49
0,75
1,18

>0,5
>0,5
>0,2

0 0,24

Leukocyturia
actual,
lg L/mL

Before
After
Change

3,61±0,14***
3,14±0,14
-0,46±0,20*

3,51±0,18***
3,35±0,13*
-0,16±0,17

0,43
1,04
1,15

>0,5
>0,5
>0,2

3,00 0,21

Leukocyturia
qualitative,
points

Before
After
Change

0,21±0,04***
0,08±0,03*
-0,14±0,05**

0,19±0,05***
0,13±0,03***
-0,06±0,06

0,38
1,19
1,04

>0,5
>0,2
>0,5

0 0,15

Note. Significant deviations from the norm as well as changes are indicated by stars (p<0,05*,
<0,01**, <0,001***).

Table 2. Comparative characteristics of the microbiota of feces

Naftussya water used for
balneotherapy

Student’s
Statistics

Reference
value

Bacteria Fresh
(n=15)

Canned
(n=14)

t p Mean Cv/
SD

Lactobacilli,
lg CFU/g

Before
After
Change

5,36±0,36***
6,85±0,35**
+1,49±0,54**

5,41±0,36***
6,37±0,35***
+0,96±0,47*

0,10
0,98
0,75

>0,5
>0,5
>0,5

8,10 0,179

Bifidobacteria,
lg CFU/g

Before
After
Change

4,76±0,32***
6,02±0,29**
+1,26±0,45**

4,91±0,29***
5,64±0,27***
+0,73±0,38

0,35
0,96
0,90

>0,5
>0,5
>0,5

6,94 0,164

Escherichia coli
common, lg CFU/g

Before
After
Change

8,18±0,07***
8,31±0,07***
+0,12±0,09

8,15±0,07***
8,26±0,07***
+0,11±0,10

0,31
0,47
0,12

>0,5
>0,5
>0,5

8,66 0,045

E. coli hemolytic,
%

Before
After
Change

29±11**
8±7
-21±15

39±12**
14±7
-25±15

0,56
0,62
0,16

>0,5
>0,5
>0,5

0 25

E. coli attenuated,
%

Before
After
Change

77±5***
50±7***
-27±8**

74±5***
65±6***
-9±7

0,42
1,63
1,70

>0,5
>0,05
>0,05

17 1,0

Klebsiela&Proteus, % Before
After
Change

19±4***
10±3**
-9±6

19±6***
11±3**
-8±6

0,03
0,16
0,06

>0,5
>0,5
>0,5

0 11

Adhering to the Truskavetsian Scientific School's analytical algorithm, the actual/raw
parameters were normalized by recalculation by the equations:

Z = 4•(V – N)/(Max – Min) = (V – N)/SD = (V/N – 1)/Cv, where
V is the actual value; N is the normal (reference) value; SD and Cv are the standard deviation
and coefficient of variation respectively.

Fig. 1 visualizes how a weekly intake of fresh Naftussya water causes an increase in the
significantly reduced content in the microbiota of feces/gut of classical probiotics [13] to the
lower normal zone. The beneficial effect is less noticeable in relation to the total content of
Escherichia coli. Instead, the marginally increased content of the E. coli strain capable of



70

hemolysis is almost completely normalized. However, the extremely high content of the E.
coli strain with weakened enzymatic activity remains in the pessimistic zone. Finally, the less
elevated content of conditionally pathogenic Klebsiela&Proteus decreases only to the upper
normal zone.

The physiological benefit of the described effects on the microbiota is confirmed by their
combination with a significant reduction of bacteriuria and leveling of leukocyturia as
markers of pyelonephritis.

Fig. 1. Profiles of normalized parameters of microbiota and leukocyturia in patients
with chronic pyelonephritis before and after a course of balneotherapy with fresh and
canned Naftussya water

The effects of Naftussya water are manifested even more clearly in direct differences
between individual final and initial values (Fig. 2). At the same time, it can be seen that the
effect of canned Naftussya water on the registered parameters is somewhat weaker, but the
differences are statistically insignificant.

Fig. 2. Profiles of changes in normalized parameters of microbiota and leukocyturia in
patients with chronic pyelonephritis after a course of balneotherapy with fresh and
canned Naftussya water



71

From the above, one gets the impression that among the organic substances of Naftussya
water [3,8] prebiotics are present.

Prebiotics are a group of biological nutrients that are capable of being degraded by
microflora in the gastrointestinal tract, primarily Lactobacilli and Bifidobacteria. When
prebiotics are ingested, either as a food additive or as a supplement, the colonic microflora
degrade them, producing short-chain fatty acids, which are simultaneously released in the
colon and absorbed into the blood circulatory system. Prebiotics boost the proliferation of gut
microbes [2,7,10].

In May 2019, the International Scientific Association for Probiotics and Prebiotics (ISAPP)
convened a panel of nutritionists, physiologists and microbiologists to review the definition
and scope of synbiotics. The panel updated the definition of a synbiotic to “a mixture
comprising live microorganisms and substrate(s) selectively utilized by host microorganisms
that confers a health benefit on the host”. The panel concluded that defining synbiotics as
simply a mixture of probiotics and prebiotics could suppress the innovation of synbiotics that
are designed to function cooperatively. Requiring that each component must meet the
evidence and dose requirements for probiotics and prebiotics individually could also present
an obstacle. Rather, the panel clarified that a complementary synbiotic, which has not been
designed so that its component parts function cooperatively, must be composed of a probiotic
plus a prebiotic, whereas a synergistic synbiotic does not need to be so. A synergistic
synbiotic is a synbiotic for which the substrate is designed to be selectively utilized by the co-
administered microorganisms. This Consensus Statement further explores the levels of
evidence (existing and required), safety, effects upon targets and implications for stakeholders
of the synbiotic concept [15].

This passage perfectly overlaps the well-known fact that about 1/3 of the mass of organic
matter in Naftussya water is the products of biotransformation by its native microbes of
organic matter leached from the aquifer [3,9,17].

The obtained results provide grounds for continuing research into the effects of canned
Naftusya water on the body of urological patients.

ACKNOWLEDGMENT

We express sincere gratitude to administration “Truskavets’ SPA” and clinical sanatorium
“Moldova” for help in conducting this investigation.

ACCORDANCE TO ETHICS STANDARDS

Tests in patients are conducted in accordance with positions of Helsinki Declaration 1975,
revised and complemented in 2002, and directive of National Committee on ethics of
scientific researches. During realization of tests from all participants the informed consent is
got and used all measures for providing of anonymity of participants.

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