Efficiency of application of sorbed probiotics in the complex therapy of pneumonia caused by SARS-CoV-2. Quality of life in the short term COVID-19

Aim. To determine the clinical efficacy and safety of the immobilized (sorbed) probiotics Bifidobacterium bifidum 1 (5x108 CFU) and B. bifidum 1 (5x107 CFU) in combination with Lactobacillus plantarum 8P-A3 (5x107 CFU) in the complex therapy of pneumonia caused by SARS-CoV-2 in adult patients without severe risk factors and their impact on health-related quality of life (QoL). Material and methods. An open, randomized prospective study included 100 patients (45 males, 55 females), aged from18 to 60 years without risk factors for severe COVID-19, with pneumonia confirmed by computed tomography and an area of lung lesion no more than 75% (moderate forms). SARS-CoV-2 RNA in nasal and oropharyngeal swabs (RT-PCR) was detected in 72% of the participants, in the rest it was highiy probable in terms of the aggregate parameters. The publication presents the results of self-assessment (94 respondents) of working capacity limitations, shortness of breath, intestinal disorders since the end of the probiotic regimen (PR: hospitalization period - B. bifidum 1, 3 capsules 2 times a day for 10 days, then after hospitalization - B. bifidum 1 in combination with L. plantarum 8P-A3 2 powders 3 times a day for 14 days) and QoL (Short Form Medical Outcomes Study: SF-36) 1 month after hospitalization. Results. At the end of PR, the ability to engage in daily activities was higher by 23.1% [95% confidence interval 5.3-37.3, OR 0.08 (0.08-0.77)]. Difficulty of breathing during exercise was less common by 29.7% [15.1-44.5%], OR 0.09 [0.02-0.40], hard stools and no bowel movements were recorded less often by 21.3% [8.5-34, 1] for 1-3 days. One month after hospitalization, the QoL of the patients receiving standard treatment was significantly reduced relative to population indicators in Russia. It was more significantly due to the psychological component of health [total measurement 38.1 (36.2-40.0)] and less significantly due to the physical component [49.5 (48.3-50.8)]. The main reasons limiting daily activities [Role Emotional (RE): 39.4 (37.4-41.4)] were decreased vitality [VT: 40.2 (38.9-041.5)], emotional depression [Mental Health (MH): 41.2 (39.4-43.0)], deficit of social contacts [Social Functioning (SF): 45.1 (43.7-46.6)]. The patients who received PR had a higher ability to carry out daily activities [RE: 57.7 (55.6-59.7)], the severity of psychological problems associated with fatigue, anxiety and depression was lower [MH: 59.8 (58.7-60.9), p<0,001]. The effect of the PR on negative perceptions of social isolation was comparatively less [SF: 53.9 (52.2-55.4)]. The QoL of the patients who additionally suffered from diarrhea in the acute period of SARS-CoV-2 pneumonia was worse in comparison with the patients without diarrhea (due to pain and inability to endure physical activity). The effects of immobilized (sorbed) probiotics to the QoL of the patients with diarrhea during the acute period of COVID-19 were most significant. Conclusion. PR had a significant positive effect on the QoL within a month after hospitalization, increasing working capacity and improving mental health, reducing the severity of psychological problems and fatigue. Additional researches are needed on the possible relationship of organic and functional gastrointestinal diseases with the progression of diarrhea in patients infected with SARS-CoV-2. No side effects of the sorbed probiotics regimen have been identified.Copyright © Eco-Vector, 2022.

Efficiency of application of sorbed probiotics in the complex therapy of pneumonia caused by SARS-CoV-2. Quality of life in the short term COVID-19

Aim. To determine the clinical efficacy and safety of the immobilized (sorbed) probiotics Bifidobacterium bifidum 1 (5×108 CFU) and B. bifidum 1 (5×107 CFU) in combination with Lactobacillus plantarum 8P-А3 (5×107 CFU) in the complex therapy of pneumonia caused by SARS-CoV-2 in adult patients without severe risk factors and their impact on health-related quality of life (QoL). Material and methods. An open, randomized prospective study included 100 patients (45 males, 55 females), aged from18 to 60 years without risk factors for severe COVID-19, with pneumonia confirmed by computed tomography and an area of lung lesion no more than 75% (moderate forms). SARS-CoV-2 RNA in nasal and oropharyngeal swabs (RT-PCR) was detected in 72% of the participants, in the rest it was highiy probable in terms of the aggregate parameters. The publication presents the results of self-assessment (94 respondents) of working capacity limitations, shortness of breath, intestinal disorders since the end of the probiotic regimen (PR: hospitalization period – B. bifidum 1, 3 capsules 2 times a day for 10 days, then after hospitalization – B. bifidum 1 in combination with L. plantarum 8P-А3 2 powders 3 times a day for 14 days) and QoL (Short Form Medical Outcomes Study: SF-36) 1 month after hospitalization. Results. At the end of PR, the ability to engage in daily activities was higher by 23.1% [95% confidence interval 5.3–37.3, OR 0.08 (0.08–0.77)]. Difficulty of breathing during exercise was less common by 29.7% [15.1–44.5%], OR 0.09 [0.02–0.40], hard stools and no bowel movements were recorded less often by 21.3% [8.5–34, 1] for 1–3 days. One month after hospitalization, the QoL of the patients receiving standard treatment was significantly reduced relative to population indicators in Russia. It was more significantly due to the psychological component of health [total measurement 38.1 (36.2–40.0)] and less significantly due to the physical component [49.5 (48.3–50.8)]. The main reasons limiting daily activities [Role Emotional (RE): 39.4 (37.4–41.4)] were decreased vitality [VT: 40.2 (38.9–041.5)], emotional depression [Mental Health (MH): 41.2 (39.4–43.0)], deficit of social contacts [Social Functioning (SF): 45.1 (43.7–46.6)]. The patients who received PR had a higher ability to carry out daily activities [RE: 57.7 (55.6–59.7)], the severity of psychological problems associated with fatigue, anxiety and depression was lower [MH: 59.8 (58.7–60.9), р<0,001]. The effect of the PR on negative perceptions of social isolation was comparatively less [SF: 53.9 (52.2–55.4)]. The QoL of the patients who additionally suffered from diarrhea in the acute period of SARS-CoV-2 pneumonia was worse in comparison with the patients without diarrhea (due to pain and inability to endure physical activity). The effects of immobilized (sorbed) probiotics to the QoL of the patients with diarrhea during the acute period of COVID-19 were most significant. Conclusion. PR had a significant positive effect on the QoL within a month after hospitalization, increasing working capacity and improving mental health, reducing the severity of psychological problems and fatigue. Additional researches are needed on the possible relationship of organic and functional gastrointestinal diseases with the progression of diarrhea in patients infected with SARS-CoV-2. No side effects of the sorbed probiotics regimen have been identified. © Eco-Vector, 2022.

Protein hydrolysates of a probiotic strain of Lactobacillus plantarum as new promising sources of inhibitors for diabetes regulatory enzymes

Diabetes is a chronic metabolic disorder characterized by persistent hyperglycaemia. It has been classified as one of the main pathologies that promotes the most severe and fatal forms of COVID-19. Although there are nowadays synthetic molecules for the regulation of this disease, their disastrous side effects still open up prospects for the search of new and healthy molecules of therapeutic interest. This study aimed at evaluating the inhibitory effect of peptides extracts produced during the cultivation of a Lactobacillus plantarum strain with probiotic properties. The peptides extracts were obtained by thermal hydrolysis of protein extracts at 70 degrees C or 80 degrees C for 10, 20 and 30 min. The potential hypoglycaemic activity was examined by testing their inhibitory effect on amylolytic enzymes. Our results showed that the studied proteins hydrolysates, when used at 0.024 mg (biomass) or 0.156 mg (growth broth), were capable of total inhibition of amylolytic enzymes activity. The biomass peptides showed higher inhibition rate compared to 95% obtained with 0.050 mg of acarbose, a potent and widely used anti-diabetic inhibitory. This study clearly shows that the investigated protein hydrolysates contain interesting inhibitory peptides that deserve further investigations for their real use as bioactive inhibitors of diabetes regulating enzymes.

The Probiotic Attributes and Anti-pseudorabies Virus Evaluation of Lactobacillus Isolates.

The emergence of pseudorabies virus (PRV) variants brings serious harm to the swine industry, and its effective treatments are limited at present. As one of the probiotics, the Lactobacillus species have beneficial characteristics of regulating the balance of intestinal flora, inhibiting the growth of pathogenic bacteria and viruses' proliferation, and improving self-immunity. In this study, Lactobacillus plantarum HN-11 and Lactobacillus casei HN-12 were selected and identified through morphology observation, Gram stain microscopy, 16S rRNA sequencing analysis, and specific amplification of the recA gene and pheS gene. All tested isolates exhibited rapid adaptation to the different conditions, excellent acid, and bile tolerance, and sensitivity to Salmonella, Staphylococcus aureus, and Escherichia coli. The antibiotic susceptibility assay displayed the isolates sensitive to most antibiotics and resistant to Lincomycin and Norfloxacin. Moreover, the supernatants of HN-11 and HN-12 inhibited PRV proliferation in ST cells. The results of animal experiments showed that supplementing the challenged mice with the supernatants of Lactobacillus isolates in advance delayed the course of the disease. PRV was detected in the heart, liver, spleen, lung, kidney, and brain tissues of dead mice in the test groups, and its copies in the lungs were significantly decreased compared with the control mice (P < 0.05). These findings proved the advantages of L. plantarum and L. casei as potential probiotic cultures, which could provide a basis for its application in microecological preparations and functional formulations.

Antiviral effect and anti-COVID-19 potential of immunobiotics

Immunobiotics, a group of probiotics, have the effect of anti-infection by regulating immune function, which can be added in in foods or used to make adjuvants or medicines (biologics). Immunobiotics can stimulate the mucosal immune system of the body, regulate innate and acquired immunity and exert non-specific anti-microbial (bacterial and viral) infection effects through oral, nasal mucosa, sublingual and other routes, but the immune regulation function of immunobiotics is species-specific. Oral administration of Lactobacillus plantarum GUANKE stimulated the increase and maintenance of SARS-CoV-2 neutralization antibodies in mice even 6 months after immunization. When L. plantarum GUANKE was given immediately after SARS-CoV-2 vaccination, the level of SARS-COV-2 specific neutralizing antibody in bronchoalveolar lavage increased by 8 times in mice, which improved the local and systematic cellular immune response to SARS-CoV-2 of mice. Clinical studies have found that immunobiotics have the auxiliary effect in the treatment of COVID-19 by mitigating the symptoms and increase the level of SARS-CoV-2 specific antibody of the patients. It is necessary to conduct research and evaluation for the appropriate guideline of immunobiotics use as erly as possible to provide a new option for the prevention and control of COVID-19.

Application of Functional and Edible Coatings and Films as Promising Strategies for Developing Dairy Functional Products-A Review on Yoghurt Case

Edible coatings and films appear to be a very promising strategy for delivering bioactive compounds and probiotics in food systems when direct incorporation/inoculation is not an option. The production of dairy products has undergone radical modifications thanks to nanotechnology. Despite being a relatively new occurrence in the dairy sector, nanotechnology has quickly become a popular means of increasing the bioavailability and favorable health effects of a variety of bioactive components. The present review describes, in detail, the various processes being practiced worldwide for yoghurt preparation, microencapsulation, and nanotechnology-based approaches for preserving and/or enriching yoghurt with biologically, and its effect on health and in treating various diseases. In the case of yoghurt, as a perfect medium for functional ingredients supplementation, different gums (e.g., alginate, xanthan gum, and gum arabic), alone or in combination with maltodextrin, seem to be excellent coatings materials to encapsulate functional ingredients. Edible coatings and films are ideal carriers of bioactive compounds, such as antioxidants, antimicrobials, flavors, and probiotics, to improve the quality of dairy food products. Yoghurt is regarded as a functional superfood with a variety of health benefits, especially with a high importance for women's health, as a probiotic. Consumption of yoghurt with certain types of probiotic strains which contain gamma-linolenic acid or PUFA can help solve healthy problems or alleviate different symptoms, and this review will be shed light on the latest studies that have focused on the impact of functional yoghurt on women's health. Recently, it has been discovered that fermented milk products effectively prevent influenza and COVID-19 viruses. Bioactive molecules from yoghurt are quite effective in treating various inflammations, including so-called "cytokine storms" (hypercytokinaemia) caused by COVID-19.

Boosting Vaccine-Elicited Respiratory Mucosal and Systemic COVID-19 Immunity in Mice With the Oral Lactobacillus plantarum.

Boosting and prolonging SARS-CoV-2 vaccine-elicited immunity is paramount for containing the COVID-19 pandemic, which wanes substantially within months after vaccination. Here we demonstrate that the unique strain of probiotic Lactobacillus plantarum GUANKE (LPG) could promote SARS-CoV-2-specific immune responses in both effective and memory phases through enhancing interferon signaling and suppressing apoptotic and inflammatory pathways. Interestingly, oral LPG administration promoted SARS-CoV-2 neutralization antibodies even 6 months after immunization. Furthermore, when LPG was given immediately after SARS-CoV-2 vaccine inoculation, specific neutralization antibodies could be boosted >8-fold in bronchoalveolar lavage (BAL) and >2-fold in sera, T-cell responses were persistent and stable for a prolonged period both in BAL and the spleen. Transcriptional analyses showed that oral application of LPG mobilized immune responses in the mucosal and systemic compartments; in particular, gut-spleen and gut-lung immune axes were observed. These results suggest that LPG could be applied in combination with SARS-CoV-2 vaccines to boost and prolong both the effective and memory immune responses in mucosal and systemic compartments, thereby improving the efficacy of SARS-CoV-2 vaccination.

Lactobacillus plantarum induces innate cytokine responses that potentially provide a protective benefit against COVID-19: A single-arm, double-blind, prospective trial combined with an in vitro cytokine response assay.

Intestinal microbiota can indirectly modulate airway physiology and immunity through the gut-lung axis. Recent microbiome studies indicate that patients with coronavirus disease 2019 (COVID-19) exhibit a specific intestinal dysbiosis that is closely associated with the disease pathophysiology. Therefore, rebalancing the intestinal microbiome using probiotics may be effective for controlling COVID-19. However, the rationale for using probiotics in COVID-19 remains unclear. In the present study, an in vitro cytokine response assay was conducted, followed by a single-arm, double-blind, prospective trial to evaluate the immunological efficacy of probiotic lactic acid bacteria against COVID-19. The present study focused on Lactobacillus plantarum (L. plantarum), Bifidobacterium longum and Lactococcus lactis ssp. lactis, which exhibit robust protective effects against infection with respiratory RNA viruses. Considering the feasibility of long-term daily intake for prophylactic purposes, healthy uninfected individuals were enrolled as subjects. Our previous pilot trial demonstrated that oral Qingfei Paidu decoction (QFPD), a Chinese herbal medicine formulated specifically against COVID-19, upregulates plasma TNF-α, IL-1ß, IL-18 and IL-8. Therefore, the present study utilized the cytokine changes induced by QFPD to define the innate cytokine index QICI [=(TNF-α) x (IL-1ß) x (IL-18) x (IL-8)/(IL-6)] as an indicator of the anti-COVID-19 immunomodulatory potential of the lactic acid bacteria. A total of 20 eligible volunteers were enrolled, 18 of whom completed the intervention. L. plantarum demonstrated a strikingly high innate cytokine index in all subjects in the in vitro cytokine response assay. In the subsequent trial, oral intake of L. plantarum significantly increased the innate cytokine index (mean fold change, 17-fold; P=0.0138) and decreased the plasma level of IL-6 (P=0.0128), a key driver of complex immune dysregulation in COVID-19, as compared with the baseline. The cytokine index increased in 16 of 18 subjects (88.9%) with considerable individual differences in the fold change (1- to 128-fold). In line with these innate cytokine changes, L. plantarum ingestion significantly enhanced the activity of natural killer cells. By contrast, oral B. longum failed to induce a significant increase in the innate cytokine index (mean fold change, 2-fold; P=0.474) as compared with the baseline. In conclusion, L. plantarum demonstrated superior QFPD-like immunomodulatory ability and mimicked the blood cytokine environment produced by early immune responses to viral infection. Daily consumption of L. plantarum as an anti-COVID-19 probiotic may be a possible option for preventing COVID-19 during the pandemic. The present study was prospectively registered in the University Hospital Medical Information Network-Clinical Trials Registry under the trial number UMIN000040479 on 22 May 2020 (https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000046202).

Potential Adjuvant Therapeutic Effect of Lactobacillus plantarum Probio-88 Postbiotics against SARS-COV-2.

In response to the ongoing COVID-19 pandemic, the global effort to develop high efficacy countermeasures to control the infection are being conducted at full swing. While the efficacy of vaccines and coronavirus drugs are being tested, the microbiome approach represents an alternative pathophysiology-based approach to prevent the severity of the infection. In the current study, we evaluated the action of a novel probiotic Lactobacillus plantarum Probio-88 against SARS-COV-2 replication and immune regulation using an in vitro and in silico study. The results showed that extract from this strain (P88-CFS) significantly inhibited the replication of SARS-COV-2 and the production of reactive oxygen species (ROS) levels. Furthermore, compared with infected cells, P88-CFS treated cells showed a significant reduction in inflammatory markers such as IFN-α, IFN-ß, and IL-6. Using an in silico molecular docking approach, it was postulated that the antiviral activity of L. plantarum Probio-88 was derived from plantaricin E (PlnE) and F (PlnF). The high binding affinity and formation of hydrogen bonding indicated that the association of PlnE and PlnF on SARS-COV-2 helicase might serve as a blocker by preventing the binding of ss-RNA during the replication of the virus. In conclusion, our study substantiated that P88-CFS could be used as an integrative therapeutic approach along with vaccine to contain the spread of the highly infectious pathogen and possibly its variants.

Mucosal IgA response elicited by intranasal immunization of Lactobacillus plantarum expressing surface-displayed RBD protein of SARS-CoV-2.

Coronavirus Disease 2019 (COVID-19) caused by a novel betacoronavirus SARS-CoV-2 has been an ongoing global pandemic. Several vaccines have been developed to control the COVID-19, but the potential effectiveness of the mucosal vaccine remains to be documented. In this study, we constructed a recombinant L. plantarum LP18:RBD expressing the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein via the surface anchoring route. The amount of the RBD protein was maximally expressed under the culture condition with 200 ng/mL of inducer at 33 °C for 6 h. Further, we evaluated the immune response in mice via the intranasal administration of LP18:RBD. The results showed that the LP18:RBD significantly elicited RBD-specific mucosal IgA antibodies in respiratory tract and intestinal tract. The percentages of CD3 + CD4+ T cells in spleens of mice administrated with the LP18:RBD were also significantly increased. This indicated that LP18:RBD could induce a humoral immune response at the mucosa, and it could be used as a mucosal vaccine candidate against the SARS-CoV-2 infection. We provided the first experimental evidence that the recombinant L. plantarum LP18:RBD could initiate immune response in vivo, which implies that the mucosal immunization using recombinant LAB system could be a promising vaccination strategy to prevent the COVID-19 pandemic.

[Efficiency of application of sorbed probiotics in complex therapy of pneumonia caused by SARS-CoV-2. Part 1. Heating clinical displays period].

AIM: To determine the clinical efficacy and safety of the sorbed probiotics Bifidobacterium bifidum 1 (5108 KОЕ) and B. bifidum 1 (5107 KОЕ) in combination with Lactobacillus plantarum 8P-А3 in the complex therapy of pneumonia caused by SARS-CoV-2 in adult patients without severe risk factors. MATERIALS AND METHODS: An open, randomized prospective study included 100 patients (45 men, 55 women), aged 18 to 60 years without risk factors for severe COVID-19 with pneumonia confirmed by computed tomography, and an area of lung lesion no more than 75% (moderate forms). SARS-CoV-2 RNA in nasal and oropharyngeal swabs (RT-PCR) was detected in 72% of the participants, in the rest it was highly probable in terms of the aggregate parameters. Diagnostics of COVID-19 and its severity, the appointment of a standard examination and treatment were carried out in accordance with the Temporary Methodological Recommendations of the Ministry of Health of Russia, version 8 of 09.03.2020. This publication presents the results of using B. bifidum 1 (3 capsules twice a day for 10 days) during the peak of clinical manifestations (in a hospital). RESULTS: In those who received sorbed B. bifidum 1, by the 10th day of treatment, the frequency of weakness was 32% lower (RR 0.55 [95% CI 0.240.73], OR 0.25 [0.110.59]); hypoosmia/dysgeusia by 22% (RR 0.42 [0.050.65], OR 0.40 [0.170.90]) and cough by 24% (RR 0.39 [0.070.60], OR 0.38 [0.170.84]). B. bifidum 1 reduced the average duration of weakness by 3 days [1.14.9], hypoosmia/dysgeusia by 3.2 days [1.35.1], cough by 1.9 days [0.43,4], dyspnea by 1.8 days [0.72.7], diarrhea by 1.7 days [0.13.5]; reduced the risk of antibiotic-associated diarrhea by 20% (RR 0.77 [0.240.93], OR 0.18 [0.050.68]). Due to the deterioration of the condition and the increase in the symptoms of respiratory failure, additional treatment was required less often by 24% (p=0.005). After the end of the intervention, the frequency of virologic debridement, levels of CRP, leukocytes, lymphocytes, platelets and the degree of lung damage on computed tomography did not statistically differ in the compared groups. No side effects of B. bifidum 1 (5108 KОЕ) have been identified. CONCLUSION: The use of sorbed B. bifidum 1 (5108 KОЕ) improved the well-being of patients without risk factors with moderate viral (SARS-CoV-2) pneumonia and reduced the duration of diarrheal syndrome in a short time. The safety profile of their use was high. More research is needed to clarify the anti-inflammatory effects of the sorbed probiotic.

A recombinant Lactobacillus plantarum strain expressing the spike protein of SARS-CoV-2.

Coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a global pandemic in the past four months and causes respiratory disease in humans of almost all ages. Although several drugs have been announced to be partially effective treatments for this disease, no approved vaccine is available. Here, we described the construction of a recombinant Lactobacillus plantarum strain expressing the SARS-CoV-2 spike protein. The results showed that the spike gene with optimized codons could be efficiently expressed on the surface of recombinant L. plantarum and exhibited high antigenicity. The highest protein yield was obtained under the following conditions: cells were induced with 50 ng/mL SppIP at 37 °C for 6-10 h. The recombinant spike (S) protein was stable under normal conditions and at 50 °C, pH = 1.5, or a high salt concentration. Recombinant L. plantarum may provide a promising food-grade oral vaccine candidate against SARS-CoV-2 infection.

Surface-Displayed Porcine IFN-λ3 in Lactobacillus plantarum Inhibits Porcine Enteric Coronavirus Infection of Porcine Intestinal Epithelial Cells.

Interferon (IFN)-λ plays an essential role in mucosal cells which exhibit strong antiviral activity. Lactobacillus plantarum (L. plantarum) has substantial application potential in the food and medical industries because of its probiotic properties. Alphacoronaviruses, especially porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), cause high morbidity and mortality in piglets resulting in economic loss. Co-infection by these two viruses is becoming increasingly frequent. Therefore, it is particularly important to develop a new drug to prevent diarrhea infected with mixed viruses in piglets. In this study, we first constructed an anchored expression vector with CWA (C-terminal cell wall anchor) on L. plantarum. Second, we constructed two recombinant L. plantarum strains that anchored IFN-λ3 via pgsA (N-terminal transmembrane anchor) and CWA. Third, we demonstrated that both recombinant strains possess strong antiviral effects against coronavirus infection in the intestinal porcine epithelial cell line J2 (IPEC-J2). However, recombinant L. plantarum with the CWA anchor exhibited a more powerful antiviral effect than recombinant L. plantarum with pgsA. Consistent with this finding, Lb.plantarum-pSIP-409-IFN-λ3-CWA enhanced the expression levels of IFN-stimulated genes (ISGs) (ISG15, OASL, and Mx1) in IPEC-J2 cells more than did recombinant Lb.plantarum-pSIP-409-pgsA'-IFN-λ3. Our study verifies that recombinant L. plantarum inhibits PEDV and TGEV infection in IPEC-J2 cells, which may offer great potential for use as a novel oral antiviral agent in therapeutic applications for combating porcine epidemic diarrhea and transmissible gastroenteritis. This study is the first to show that recombinant L. plantarum suppresses PEDV and TGEV infection of IPEC-J2 cells.