THE USE OF PERSONALIZED PHARMABIOTICSAS AN APPROACH TO THE REHABILITATIONOF POST-COVID PATIENTS

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https://doi.org/10.15407/microbiolj86.04.064  L.S. YUSKO1*, S.A. BURMEI1, I.S. LEMKO2, A.I. KRASTANOV3, N.V. BOYKO1  1

Department of Medical and Biological Disciplines,  State Higher Educational Institution «Uzhhorod National University»,  3 Narodna Square, Transcarpathian region, Uzhhorod, 88000, Ukraine  2Government Institution «Th e Scientifi c-Practical Medical Centre «Rehabilitation» Health Ministry of Ukraine»,  10 Velikokamyana Str., Transcarpathian region, Uzhhorod, 88000, Ukraine  3Department of Biotechnology, University of Food technologies,  26 Maritza Blvd, Plovdiv, 4002, Bulgaria  * Author for correspondence; e-mail: lesyus@ukr.net 

THE USE OF PERSONALIZED PHARMABIOTICS  AS AN APPROACH TO THE REHABILITATION  OF POST-COVID PATIENTS 

Successful application of defi ned pro- and/prebiotic preparations for the prevention and treatment of viral respiratory    infections is confi rmed by meta-analyses and numerous clinical trials. To date, the protocols for the rehabilitation of pa-  tients with post-COVID conditions, an integral part of which is the restoration of the balance of gut microbiota along with    nutritional supportу, are widely developed and accepted. Purpose. To investigate the effi cacy of individually prescribed  pharmabiotics for targeted correction of the nasal and gut microbiota of post-COVID-19 patients in combination with  aerosol inhalations. Methods. Th e post-COVID-19 patients were referred to recover using the rehabilitation facilities. In  addition to the basic treatment complex, the patients were off ered haloaerosol therapy with an additional prescription  of individually selected pharmabiotics. Results. Th e use of individually prescribed pharmabiotics in combination with  aerosol inhalation enabled therestoration of Lactobacillus spp. balance and reduction in the number of opportunistic  microbiota in the gut. Th us, the personalized rehabilitation approach led to signifi cantly improved local immune response    in post-COVID-19 patients. Conclusions. Th e data obtained provide supportive evidence of the effi cacy of aerosol inha-  lations and personalized pharmabiotics Lactobacillus rhamnosus S25 and L. plantarum A combined application in the    directed modulation of the microbiome and targeted correction of local immunity in post-COVID patients. Restoring the  balance of the patient’s’ oral and gut microbiota should be an integral part of the post-COVID patient’s rehabilitation.  Keywords: COVID-19, post-COVID rehabilitation, gut-lung axis, pharmabiotics.    

COVID-19 is a potentially severe acute respira-  tory infection caused by the SARS-CoV-2 coro-  navirus. To date, there is no reliable data on the    pathogenesis of COVID-19. Scientists believe    that the virus enters the cell by binding to an-  giotensin-converting enzyme 2 (ACE2) recep-  tors, which the virus uses as an «entry gate» (de    Oliveira et al., 2021; Troisi et al., 2021). Rather  than being limited to the lower respiratory    tract, severe acute respiratory syndrome infec-  tion caused by SARS-CoV-2 has been proven to    spread to other organs, namely the gastrointes-  tinal tract (multi-tissue infection). In particular,    some manifestations of SARS-CoV-2 infection,  such as the ability of SARS-CoV-2 to infect and  replicate in small human intestinal enterocytes,  detection of viral RNA in fecal samples and the    changed composition of the gut microbiota un-  der SARS-CoV-2, indicate signifi cant damage to    the gastrointestinal tract during the disease (de  Oliveira et al., 2021; Troisi et al., 2021; Lamers et    al., 2020). Host-microbiota interactions are bidi-  rectional, complex, and potentially modulate the    development and functioning of the innate and  adaptive immune systems. Signals from the gut  microbiota can tune immune-mediated cells to  pro-infl ammatory (T helper cells type 17; Th 17)  and anti-infl ammatory (regulatory T cells; Tregs)  responses, determining susceptibility to various  diseases. Th ere is a lot of data indicating that the  composition of intestinal microbiota is directly  related to disease severity and concentration of  cytokines and infl ammatory markers in plasma.  Moreover, the composition of gut microbiota of  recovered patients remains signifi cantly altered  compared to people who were not infected with    SARS-CoV-2, which may infl uence the develop-  ment of the post-COVID syndrome (Yeoh et al.,    2021). For example, the studies conducted by a  group of scientists from China demonstrate that  intestinal microbiota in patients with COVID-19  is characterized by an increase in the number of    Streptococcus, Clostridium, Lactobacillus, and Bi-  fi dobacterium and a decrease in the number of    Bacteroidetes, Roseburia, Faecalibacterium, Co-  prococcus, and Parabacteroides compared to pa-  tients with seasonal fl u and healthy people (Fer-  reira et al., 2020). Th us, it is most likely that a    healthy gut microbiota, which protects the lungs    and vital organs from an exaggerated immune re-  sponse, can counteract the coronavirus infection.    In 2001, a joint consultation of the Food and  Agriculture Organization (FAO) and the World  Health Organization (WHO) experts agreed on    a specifi c defi nition of probiotics as «live micro-  organisms that when administered in adequate    amounts confer a health benefi t on the host.»  Over the past few decades, greater research of  the gut-brain-, gut-lung- and gut-liver-axes has  expanded our understanding of the microbiome  and fostered to the advancement of pharmabiotics.  Pharmabiotics refer to live microbes administered    to patients to treat a disease. Pharmabiotics diff er-  from prebiotic and probiotic products as they are    validated through clinical trials with safety and ef-  fi cacy endpoints for a given indication and follow    the same regulatory pathway to approval as other  novel drugs (LeBegue et al., 2020).    In addition, with the development of predic-  tive, preventive, and personalized medicine (P3    medicine), personalized pharmabiotics began to  be used. Th e term «personalized pharmabiotics»    means that pharmabiotics are personally (indi-  vidually) selected for a given patient based on    the state of his/her microbiota.  Th e number of studies assessing the impact    of pro- and prebiotics use on frequency, dura-  tion, and severity of viral respiratory infections    in humans is signifi cantly increasing. Th e poten-  tial of probiotics use is supported by experimen-  tal studies, meta-analyses, and clinical trials on    other coronaviruses and such viruses as infl u-  enza viruses, rhinoviruses, and respiratory syn-  cytial virus (Waki et al., 2014; Luoto et al., 2014;    Turner et al., 2017; Kumar et al., 2010; Liu et al.,    2020; Wang et al., 2019). Probiotics such as Lac-  tobacillus and Bifi dobacterium have been proven    to restore human health by eliminating patho-    66 ISSN 1028-0987. Microbiological Journal. 2024. (4)  L.S. Yusko, S.A. Burmei, I.S. Lemko, A.I. Krastanov, N.V. Boyko  genic microorganisms and regulating immune  responses in intestinal epithelial cells (Kurian  et al., 2021). Finally, although the mechanisms  involved in SARS-CoV-2 infection are not yet  fully understood, there is research evidence that    directly links gut microbiota to COVID-19, con-  fi rming the likely role of probiotics in both the    prevention and treatment of COVID-19 (Zuo et  al., 2020; Liu et al., 2021; Yeoh et al., 2021; Gou  et al., 2020; Mańkowska-Wierzbicka et al., 2023).  According to medical statistics, contracting  COVID-19 oft en results in a number of specifi c    problems in various body systems. Hence, com-  plex rehabilitation following coronavirus infec-  tion should include a whole set of measures.    Recently, in addition to the terms «post-COVID  syndrome» and «chronic COVID-19,» a new  defi nition has appeared in the foreign medical  literature — «long COVID». People unable to get  rid of the coronavirus infection consequences    for a long time are called «long haulers» in medi-  cal slang.    Since the virus causes systemic disorders in  the body, it is absolutely necessary to not only    treat patients correctly but also manage the im-  balance that occurs aft er the illness. Restorative    therapy is needed to alleviate the post-COVID  syndrome and eventual recovery, as dysbiotic gut  microbiome may in turn contribute to immune  and health-related problems aft er COVID-19.    Everyone needs recovery aft er a coronavirus in-  fection, regardless of the severity of the disease.    However, the scope of restorative and rehabili-  tation programs should rely on an individual    approach to each patient. Now, most countries    across the world off er protocols for the reha-  bilitation of patients with post-COVID condi-  tions including restoration gastrointestinal tract    (GIT) microbiota’ balance and patient nutrition-  al support (Ferreira et al., 2020; Zuo et al., 2020).    However, in Ukraine, this issue remains open  and therefore extremely relevant.  Th e purpose of our work was to investigate    the possibility of using personalized pharmabi-  otics for targeted correction of the microbiota of    postcovid-19 patients in combination with aero-  sol inhalations.    Materials and Methods. A total of 100 people  who had contracted COVID-19 and treated at  the State Institution «Rehabilitation Scientifi c  and Practical Medical Center of the Ministry of  Health of Ukraine» took part in the study. All  patients signed the «Informed Consent of the  Study Participant». During the performance of    the scientifi c research submitted for examina-  tion, compliance with safety rules was ensured    by the principles of the Declaration of Helsinki,  adopted by the General Assembly of the World  Medical Association (1964—2016), the Council  of Europe Convention on Human Rights and  Biomedicine (1997), the European Convention  for the Protection of Vertebrate Animals Used  for Research and Other Scientifi c Purposes  (1986), the relevant provisions of the WHO, the    International Council of Medical Scientifi c So-  cieties, the International Code of Medical Eth-  ics (1983) and the relevant laws of Ukraine (Th e    Ethics Committee of the State Higher Educa-  tional Establishment «Uzhhorod National Uni-  versity») approved the study protocol No. 9/7 on    07/06/20223).  Taking into account the peculiarities of the    convalescence period, apart from the basic treat-  ment complex, patients were also off ered halo-  aerosol therapy (HAT) with additional prescrip-  tion of individually selected pharmabiotics.    All clinical bacterial strains used in this study    are preserved in the collection of microorgan-  isms at the Research Development and the Edu-  cational Center of Molecular Microbiology and    Mucosal Immunology at the Uzhhorod National  University.    Th e basic treatment complex included meth-  ods of equipment-based physiotherapy and the    necessary medical treatment:  ■ singlet oxygen therapy;  ■ electrophoresis with a solution of heparin or  lidase on the rib cage;    ISSN 1028-0987. Microbiological Journal. 2024. (4) 67  Th e Use of Personalized Pharmabiotics as an Approach to the Rehabilitation of Post-COVID Patients    ■ individual inhalations according to indica-  tions (broncholytics, dekasan, phytoncides);    ■ aeroionotherapy;    ■ other physiotherapeutic procedures as indi-  cated;    ■ chest massage as indicated;  ■ exercise therapy.  Th e treatment course lasted 21 days.    Aft er completing the treatment course, pa-  tients were additionally prescribed individually    selected pharmabiotics. Th e course of pharma-  biotics lasted seven days. All the patients gave    informed consent.    Th e intensity of the local immunity of mu-  cous membranes (oral and intestinal), including    specifi c antibodies to COVID-19, secretory im-  munoglobulin A, and infl ammatory and anti-  infl ammatory cytokines, was studied in patients.    Personalized pharmabiotics were selected aft er    gut and upper respiratory tract microbiota stud-  ies in order to modulate microbiota and directly    correct local immunity.    Quantitative analysis of microbiota was con-  ducted using the routine cultural method.    Saliva samples diluted in 0.9% sodium chlo-  ride and faecal samples in MRS broth aft er se-  rial dilutions were plated at the amount of 10    μL of each dilution (101, 102, 104, 106, 108, 1010)  on appropriate nutrient media, in particular  MRS agar, Bifi dobacterium agar, meat-peptone  agar (MPA), 5% blood agar (BA), Sabouraud  medium, Endo medium, Bile medium, Strepto    medium, Clostridial agar, and selective staphy-  lococcal medium with mannitol. All dishes were    incubated for 24—48 hours at 37±1 °C under  appropriate cultivation conditions (aerobic and  anaerobic).  Morphological and tinctorial properties of  microorganisms were determined using the  Gram method by digitizing the swab.  Th e state of the upper respiratory tract (URT)    and gastro-intestinal tract (GIT) mucous mem-  branes microbiocenosis of the patients was eval-  uated using the persistence index (C%) and the    frequency index (Рі):  С% = р/Р×100,    where C% is the persistence index; p is the num-  ber of samples containing the tested strain of    bacteria; P is the total number of samples that  contain all isolated strains of bacteria.  Рі = А/В,    where A is the number of species strains; B is the  total number of strains.  Immune parameters including IgA, IL-6, and  TNF-ά were measured using an immunosorbent    system Labor Diagnostika Nord (Germany) ac-  cording to the manufacturer’s instructions.    For microbiome modulation of local immunity  in post-COVID patients, a personalized selection    of pharmabiotics was conducted. Deposited, au-  thorial strains of lactobacilli, namely Lactobacil-  lus bulgaricus S6, L. delbrueckii subsp. bulgaricus    S19, L. rhamnosus S25, L. plantarum A, L. bulgari-  cus А6, and L. bulgaricus А 22 were selected as    pharmabiotics based on their biological proper-  ties. Th ese strains were isolated from traditional    dairy products and endemic plants in high moun-  tain, ecologically clean areas of the Rhodope    Mountains in Bulgaria (Ilyazova et al., 2022).  Personalized selection of pharmabiotics was  carried out by co-cultivation of isolated from  post-COVID patient’s microorganisms and    strains of probiotic lactic acid bacteria. Co-cul-  tivation was conducted in 96 well polystyrene    microplates. 100 μL of isolated microorganisms’  suspension was plated into the wells and 100  μL of probiotic cultures suspension was added.  Suspensions were prepared in meat-peptone  broth (MPB) (isolated microorganisms) and    MRS broth (probiotic cultures) with a McFar-  land turbidity of 0.5 using an electronic device    Densi-La-Meter (PLIVA-Lachema Diagnostika,    Czech Republic), which corresponds to the con-  centration of 1.5·108 CFU/mL. Microplates with    suspensions were incubated at 37 ± 1°C for 24  hours. Aft er that, aliquots of 10 μL were plated    

on appropriate agarized nutrient media using  the streak culture method and incubated for 24  hours at 37 ± 1°C. Evaluation of the antagonistic  properties of the above-mentioned lactobacilli    strains in relation to isolated opportunistic mi-  croorganisms was carried out.    Th e study of microbiota state and intensity of  mucous membranes (URT, GIT) local immunity  was conducted in three stages:  1 — during rehabilitation;  2 — aft er the treatment complex before taking  pharmabiotics;  3 — aft er taking pharmabiotics.  Th e basic statistical analysis of the data was  conducted applying the well-known methods of    descriptive statistics and statistical inference, us-  ing the Anaconda — Python environment and    the OriginPro soft ware package of OriginLab for  numerical data analysis. Data correlation analysis    and methods of comparison of two or more de-  pendent or independent groups of data using the    Mann-Whitney, Wilcoxon, and Kruskel-Wallis  criteria were applied. Th e results were considered  statistically signifi cant at p<0.05 level.    Results. Personalized selection of pharma-  biotics. Th e results of individual selection of    pharmabiotics indicated that of six lactobacilli  only two strains, namely L. rhamnosus S25 and  L. plantarum A, were the most eff ective for target  microbiome modulation. Th erefore, taking into    account individual (personal) state of microbio-  ta, the course of pharmabiotics for post-COVID    patients included taking L. rhamnosus S25 or/  and L. plantarum A strains.    Staphylococcus aureus  Bacillus spp.  Escherichia coli, lac  +  Escherichia coli, lac–  Clostridium spp.  Klebsiella oxytoca  Pseudomonas spp.  Lactobacillus spp.  Staphylococcus spp.  Enterococcus faecalis  Candida spp.  Streptococcus spp.  Actinomyces spp.  Proteus spp.    0  2  4  6  8  10    log10(Microbial concentration, CFU/g)    Before treatment  Treatment, aerosol inhalations  Treatment, aerosol inhalations and pharmabiotics    Fig. 1. Th e composition    of the intestinal micro-  biota of post-COVID    patients before and aft er  the treatment    ISSN 1028-0987. Microbiological Journal. 2024. (4) 69  Th e Use of Personalized Pharmabiotics as an Approach to the Rehabilitation of Post-COVID Patients  Study of intestinal microbiota. In patients    who had contracted COVID-19, the key intesti-  nal microbiome representatives, namely Staphy-  lococcus spp. (including S. aureus), Streptococcus    spp., Candida spp., Enterococcus faecalis, and Ac-  tinomyces spp. were revealed in increased quan-  tities compared to the reference levels (Fig. 1).    Th e strains of Escherichia coli were identifi ed in  signifi cantly lower amount (5.3 ± 0.8 log10 CFU/  mL) compared to the normal level (107—108  CFU/mL). A decrease in the number of typical  Escherichia coli down to 105—106 CFU/mL or an  increase to 109—1010 CFU/mL is considered the  fi rst stage of microbiological disorders. At the  same time, there was a signifi cant decrease in  the number of Lactobacillus spp. (5.9 ± 2.9 log10  CFU/g) compared to the reference levels (106—  108 CFU/mL). Th e number of Bifi dobacterium  spp. was even lower than the detection limit.  As for the state of GIT mucous membranes    microbiocenosis of the patients, the highest val-  ues of detection frequency indicators and per-  sistence index were demonstrated by Staphylo-  coccus spp. (C% — 100%; Рі — 0.18) (Table 1).    Th e lowest rates of persistence and detection fre-  quency were characteristic of Proteus spp. repre-  sentatives (C% — 20%; Рі — 0.05). Lactobacillus    spp. had a high detection frequency, but their  persistence index was low (C% — 100%; Рі —  0.09). For E. coli lac+, the detection frequency  was 80% and persistence index was 0.07.    Aft er aerosol inhalations and the use of person-  alized pharmabiotics, the species composition of    gut microbiota remained practically unchanged.  However, the use of personalized pharmabiotics    made it possible to restore the balance of Lacto-  bacillus spp. (6.2 ± 2.9 log10 CFU/g) and reduce    the number of key opportunistic microbiota rep-  resentatives to normal values (Fig. 1).    Detection frequency indicators of Lactobacil-  lus spp. and E. coli lac+ increased to 0.13 and    0.09 compared to the data obtained before the  therapy (Table 1).  Study of oral microbiota. Th e oral microbiota  of the post-COVID patients was represented by  the excessive number of key microorganisms  such as Staphylococcus spp. (including S. aureus),  Clostridium spp., Actinomyces spp., Streptococcus  spp., and Candida spp. (Fig. 2). Aft er the use of    aerosol inhalations, the number of these condi-    

tionally pathogenic microorganisms was signifi -  cantly decreased but was still higher than normal    values. A decrease in the number of Staphylococ-  cus spp. from 7.4 ± 1.1 log10 CFU/mL to 6.5 ±    ± 1.0 log10 CFU/mL was revealed, as well as a de-  crease in the number of Candida spp. from 7.6 ±    ± 0.9 log10 CFU/mL to 5.9 ± 0.8 log10 CFU/mL,  and Actinomyces spp. from 6.0 ± 0.6 log10  CFU/mL to 5.2 ± 0.6 log10 CFU/mL. However,  no signifi cant diff erences were found in the  amount of Clostridium spp., as the number of  this anaerobic bacterium remained at the level    of 6.0 ± 0.2 log10 CFU/mL. Aft er the personal-  ized pharmabiotics treatment course, the num-  ber of Staphylococcus spp. (including S. aureus),    Actinomyces spp., Streptococcus spp., and Can-  dida spp.) decreased to the normal values. Th e    number of Enterococcus spp. decreased to the    detection limit. At the same time, there was a    tendency to increase the number of Lactobacil-  lus spp. (5.7 ± 1.2 log10 CFU/mL) compared to    that during treatment (5.0± 1.0 log10 CFU/mL).  Analysis of the obtained data showed that  Staphylococcus spp. and Streptococcus spp. were    the dominant representatives of the oral micro-  biota. Th ey demonstrated the highest values of    the detection frequency index (Pi — 0.12) and  persistence index (С — 100%) at the time of  treatment (Table 2).  Aft er the use of aerosol inhalations together    with personalized pharmabiotics, several chang-  es in oral microbiota were detected. Th e detec-  tion frequency of such bacteria as Staphylococcus    spp. (C% — 85%; Ri — 0.08), Clostridium spp.  (C% — 20%; Ri — 0.01), Candida spp. (C% —  35%; Ri — 0.02), and Actinomyces spp. (C% —    Staphylococcus spp.  Escherichia coli, lac  +  Streptococcus spp.  Staphylococcus aureus  Clostridium spp.  Lactobacillus spp.  Candida spp.  Enterococcus spp.  Actinomyces spp.    0  2  4  6  8  10    log10(Microbial concentration, CFU/g)    Before treatment  Treatment, aerosol inhalations  Treatment, aerosol inhalations  and pharmabiotics    Fig. 2. Th e oral micro-  biota composition of the    post-covid patients be-  fore and aft er the treat-  ment    ISSN 1028-0987. Microbiological Journal. 2024. (4) 71  Th e Use of Personalized Pharmabiotics as an Approach to the Rehabilitation of Post-COVID Patients    10%; Ri — 0.01) was reduced. At the same time,  the persistence index and detection frequency  index of Lactobacillus spp. and E. coli, lac+ were  increased (C% — 55%, Ri — 0.09; C% — 25%,  Ri — 0.05, respectively).    Study of immunological parameters. Immu-  nomodulation includes natural and therapeutic    processes aimed at modifying the immune re-  sponse. In all patients who had contracted COV-  ID-19 the level of IgA was within the lower range    of the norm — 0.7—0.83 pg/mL on average ver-  sus a norm of 0.7—4 mg/mL (Table 3). However,    an increase in the IgA level (2.14—2.56 mg/mL)  was observed aft er the use of aerosol inhalations  and personalized pharmabiotics.  Th e rate of IL-6 in post-COVID patients    ranged from 7.5 to 9.41 pg/mL, which was signif-  icantly higher than the normal rate of 1.5 to 7.0    pg/mL. Th is indicates an increase in IL-6 produc-  tion due to the stress factor, SARS-CoV-2 in this    case. As known, IL-6 stimulates macrophages,  which trigger active infl ammatory processes.  Th eir excessive amount can damage healthy  cells. Some studies of early cases of COVID-19 in  China indicate that the immune response to the  coronavirus may be very active: in patients with  severe symptoms, elevated levels of cytokines,  particularly IL-6, were found (Troisi et al., 2021).  Th e so-called cytokine storm occurs. Th erefore, it  is extremely important to achieve normalization  of the serum levels of the acute phase proteins as  soon as possible in order to avoid damage to lung  tissues due to an autoimmune reaction.    Aft er a course of aerosol inhalations and person-  alized pharmabiotics, the IL-6 indicator was 2.32—    5.58 pg/mL, which is within the normal range.    In our studies, the level of TNF-α in all pa-  tients was within the range of 5.88—5.9 pg/mL    (reference values 0—13 pg/mL), i.e. it had no di-  agnostic value.    

Th us, the complex use of aerosol inhalations    and personalized pharmabiotics made it pos-  sible to improve the state of mucous membrane    microbiota of URT and GIT of the patients who  had contracted COVID-19.  Discussion. Mucosal surfaces, particularly    those of the lungs and intestines, play an impor-  tant role in modulating immune responses, there-  by reducing the number of pathogens and pre-  venting excessive infl ammation or tissue damage.    Th e formation of an adaptive immune response    on mucous membranes involves certain peculiar-  ities. Th e main antigen-presenting cells of muco-  sal membrane are B-lymphocytes and mast cells.    Local synthesis of IL-4 is observed, which causes    a Tx2-dependent humoral response, mainly ac-  companied by the formation of IgA antibodies.    Th e latter can penetrate through the epithelial  cells to the surface of mucous membranes, where  they interact with pathogens and neutralize them,  thereby contributing to their rapid elimination.    However, the harmonious setting of local im-  munity directly depends on the balance of the    so-called «local microbiota»: violation of mucosal    tolerance together with a dysbiotic state can con-  tribute to infection and progression of infections,    including those caused by SARS-CoV-2. Since    under COVID-19, respiratory and gastrointesti-  nal mucosal membranes are aff ected and changes    in microbiota composition occur, it is plausible  to suggest that targeted correction based on the  modulation of the gut-lung axis and restoration    of eubiosis may be an important therapeutic ap-  proach to limiting COVID-19 harmful eff ects.    Recently, there has been increasing evidence  that the use of pharmabiotics and nutraceutical    support demonstrates anti-infl ammatory prop-  erties and improves the body’s immunomodula-  tory properties (Ferreira et al., 2020; Kumar et    al., 2010; Wei et al., 2021).  Numerous probiotics have been shown to  be benefi cial under coronavirus infections, but  evidence detailing their eff ectiveness in treating  COVID-19 in particular is limited (Ferreira et    al., 2020). L. plantarum Probio-38 and L. salivar-  ius Probio-37 can inhibit transmissible gastro-  enteritis coronavirus (Luoto et al., 2014; Kumar    et al., 2010). Probiotic E. faecium NCIMB 10415  has been approved in the European Union as a  feed additive for young piglets for the treatment    of transmissible coronavirus gastroenteritis (Ku-  mar et al., 2010; Wei et al., 2021). Recombinant    IFN-λ3-anchored L. plantarum can inhibit swine  gastroenteritis caused by coronavirus in vitro  (Liu et al., 2020; Averina, 2017). However, the  clinical utility of probiotics in human infections    caused by SARS-CoV-2 requires further evalua-  tion (Ferreira et al., 2020; Han et al., 2021).    Registered clinical trials of probiotics under    COVID-19, mostly Lactobacillus and Bifi dobacte-  rium-Lactobacillus mixtures, are ongoing; hence,    the preventive or therapeutic role of probiotics in  such patients may be elucidated in the near future.    So, representatives of Staphylococcus spp. (in-  cluding S. aureus), Streptococcus spp., Clostrid-  ium spp., E. faecalis, K. oxytoca, Candida spp.,    and Actinomyces spp. prevailed in mucous mem-  brane microbiota of URT of the patients who    had contracted COVID-19; in GIT microbiota,    Staphylococcus spp. (including S. aureus), Strep-  tococcus spp., Clostridium spp., and Candida    spp. had the largest numbers.    Aft er the use of aerosol inhalations and person-  alized pharmabiotics, the qualitative composition    of URT and GIT mucous membrane microbiota  of remained practically unchanged. However, a    signifi cant decrease in the amount of opportunis-  tic microbiota to normal levels was revealed. Th e    frequency of detection of Enterococcus spp. and    Candida spp. decreased to 0.02, whereas Clostrid-  ium spp. and Actinomyces spp. — to 0.01.    Th e use of aerosol inhalations in combination  with personalized pharmabiotics signifi cantly  improved the local immune response state of the    patients who had contracted COVID-19, as evi-  denced by an increase in IgA (from 0.7—0.83 to    2.14—2.56 mg/mL) and a decrease in IL-6 (from  7.5—9.41 to 2.32—5.58 pg/mL).    

Th us, the obtained results prove the eff ective-  ness of aerosol inhalations and the use of per-  sonalized pharmabiotics L. rhamnosus S25 and    L. plantarum A in modulating the microbiome  and targeted correction of local immunity in  post-COVID patients. Restoring the balance of  URT and GIT mucous membrane microbiota  of the patients who have contracted COVID-19  should be an integral part of the rehabilitation  of post-COVID patients.   

Conclusions.

Th e d ata obtained provide sup-  portive evidence of the effi cacy of the combined    application of aerosol inhalations and person-  alized pharmabiotics L. rhamnosus S25 and    L. plantarum A in the directed modulation of  the microbiome and targeted correction of local  immunity in post-COVID patients.  Restoring the balance of the patient’s oral and  gut microbiota should be an integral part of the  post-COVID patient’s rehabilitation.  Sources of Funding. Th e authors received no  fi nancial support for their study.  Confl ict of Interest. Th e authors declare that  they have no confl ict of interest.  Ethical approval. Th e bioethics committee    of State Higher Educational Institution «Uzh-  horod National University», protocol No 9/7 of    07.06.2023.   

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Бойко1  1 Кафедра медико-біологічних дисциплін,  Державний вищий навчальний заклад «Ужгородський національний університет»,  площа Народна, 3, Закарпатська область, Ужгород, 88000, Україна  2 Державна установа «Науково-практичний медичний центр «Реабілітація»  Міністерства охорони здоров’я України»,  вул. Великокам’яна, 10, Закарпатська область, Ужгород, 88000, Україна  3 Кафедра біотехнології, Університет харчових технологій,  бульвар Маріца, 26, Пловдив, 4002, Болгарія  ВИКОРИСТАННЯ ПЕРСОНАЛІЗОВАНОЇ ФАРМАКОТЕРАПІЇ  ЯК ПІДХОДУ ДО РЕАБІЛІТАЦІЇ ПОСТКОВІДНИХ ПАЦІЄНТІВ    Успішне застосування визначених про-/i пребіотиків для профілактики та лікування вірусних респіратор-  них інфекцій підтверджено мета-аналізом та численними клінічними дослідженнями. На сьогодні розро-  блено та прийнято протоколи реабілітації пацієнтів із постковідними станами, невід’ємною частиною яких    є нутритивна підтримка та відновлення балансу мікробіоти кишківника. Мета. Дослідити ефективність    індивідуально призначених фармакобіотиків для цілеспрямованої корекції назальної та кишкової мікро-  біоти хворих, які перехворіли на COVID-19, у поєднанні з аерозольними інгаляціями. Методи. Пацієнтам,    які перенесли COVID-19, крім основного лікувального комплексу, була запропонована галоаерозольтерапія    з додатковим призначенням індивідуально підібраних фармабіотиків. Результати. Застосування індивіду-  ально призначених фармабіотиків у поєднанні з аерозольними інгаляціями дозволило відновити баланс    Lactobacillus spp. і зменшити кількість умовно-патогенної мікробіоти кишківника. Персоналізований реа-  білітаційний підхід сприяв значному покращенню місцевої імунної відповіді в пацієнтів після COVID-19.    Висновки. Отримані дані доводять ефективність аерозольних інгаляцій і персоналізованих фармакобіо-  тиків Lactobacillus rhamnosus S25 і L. plantarum A у комбінованому застосуванні для спрямованої модуляції    мікробіоти та корекції місцевого імунітету в пацієнтів, які перехворіли на COVID-19. Відновлення балансу    мікробіоти верхніх дихальних шляхів та кишківника має бути невід’ємною частиною реабілітації постко-  відних пацієнтів.    Ключові слова: COVID-19, постковідна реабілітація, вісь кишечник-легені, фармакотерапія.