DO PROBIOTICS HAVE A FUTURE IN NEONATOLOGY? (ANALYSIS OF THE LATEST DATA. PART 3)

The issue of feasibility and effectiveness of probiotics use in newborns is still discussable. A position letter of the Committee on nutrition of the European Society for Pediatric NutritionGastroenterology, Hepatology, and(ESPGHAN) and the Working group of the ESPGHAN on probiotics and prebiotics issues was published in May 2020 in “Pediatric Gastroenterology and Nutrition” magazine as for the use of probiotics in premature newborns. The third part of the literature continues with the position paper and the results of many randomized controlled clinical trials of probiotics. The article considers the answers to 6 clinical questions posed by the working group of the Committee to assess the feasibility of use, routes of administration, dosage and duration of use, as well as the effectiveness and quality of probiotic drugs. It was proved that probiotics, in general, could decrease the level of necrotising enterocolitis, sepsis, and mortality. On the other hand, an increasing number of commercial products containing probiotics of non optimal quality are available. In addition, a large number of departments in the world regularly suggest probiotic supplements as a treatment standard despite the absence of any solid evidence. Moreover, the emphasis was placed on issues of safety of probiotic supplements for premature newborns. Guarantee of quality of probiotic product is deliverance of probiotic strains by transfer genes of resistance to antibiotics, the ability to regularly detect sepsis while using probiotics.

Effect of “EM® PROBIOTIC FOR BEES” on the dynamics viability of bee in an entomological cage experiment

At present, Ukraine is one of the first honey producing country in Europe. Around 100 thousand tonnes of honey are produced annually in Ukraine, therefore, maintaining the health of the bee colonies is an important issue. The use of different groups of drugs for the prevention of bee diseases is strictly controlled, now the known alternatives to antibiotics are probiotics. The micro-organisms in these preparats are able to synthesise vitamins and amino acids necessary for the growth and development of bees, which in turn activates the immunocompetent cells of the insects and prolongs their life. The immunomodulatory and immunostimulatory ability of such supplements of “beneficial” microorganisms is known, both in veterinary and human medicine. The use of such remedies is therefore promising in the field of modern beekeeping. “EM® PROBIOTIC FOR BEES” is a biopreparation that positively influences the microbiological environment of insects, as the preparation is based on Effective Microorganisms®. The ability of this probiotic to influence the dynamics of life span of Ukrainian steppe bees in a wooden entomological cage experiment (in vivo) was the main objective of the experiment. The effect of the probiotic on bee viability was determined by daily analysis and counting the number of dead insects. The study involved the settlement of bees in a wooden entomological cages and the creation of optimal conditions for their keeping (at a temperature of +24 – +25 °C and a humidity of 50–70 %). The product was diluted with buckwheat honey syrup solution and sugar syrup solution at concentrations of 5 %; 2.5 %; 1.25 %; control groups of bees received native solutions of the sugar syrup and buckwheat honey syrup. The analysis of the results shows a positive effect of “EM® PROBIOTIC FOR BEES” on the longevity of Ukrainian steppe worker bees of the winter generation in a entomological cage experiment. A beneficial effect of the probiotic product diluted in sugar syrup at concentrations of 1.25 % to 5 % has been detected on the bee organism, which increased their lifespan. When the product was diluted with buckwheat honey syrup, the best longevity of the insects was recorded at a concentration of 1.25 % compared to the control group of bees. The coefficient of average life expectancy of bees indicates the predominance of sugar syrup as a solvent for this probiotic compared to buckwheat honey syrup under laboratory conditions.

DO PROBIOTICS HAVE A FUTURE IN NEONATOLOGY?

The issue of feasibility and effectiveness of probiotics use in newborns is still discussable. A position letter of the Committee on nutrition of the European Society for Pediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) and the Working group of the ESPGHAN on probiotics and prebiotics issues was published in May 2020 in “Pediatric Gastroenterology and Nutrition” magazine as for the use of probiotics in premature newborns. It stated that over 10,000 premature newborns from all over the world had taken part in random controlled probiotics tests. It was proved that probiotics, in general, could decrease the level of necrotizing enterocolitis, sepsis, and mortality. But the question of choice of microorganism strains, dosing, and duration of medication course remains open. On the other hand, an increasing number of commercial products containing probiotics of non optimal quality are available. In addition, a large number of departments in the world regularly suggest probiotic supplements as a treatment standard despite the absence of any solid evidence. According to the data of the latest meta-analysis, effectiveness in decreasing mortality and incidence was found in the minority of investigated strains or combinations. In the position letter authors wanted to give advice which specific strains can be potentially used and which cannot. Moreover, the emphasis was placed on issues of safety of probiotic supplements for premature newborns. Guarantee of quality of probiotic product is deliverance of probiotic strains by transfer genes of resistance to antibiotics and the possibility of regular identification of probiotic sepsis. There is a conditional recommendation (with a low credibility of evidence) to provide eitherLactobacillus rhamnosus GG ATCC53103, or combination of Bifidobacterium infantis Bb-02, Bifidobacterium lactis Bb-12, and Streptococcus thermophilus TH-4 for decreasing a risk of the necrotizing enterocolitis development subject to all issues of safety.

Development of the composition and technology of a probiotic fermented milk product with the addition of a plant ingredient

The research work reflects the problems of baby food at the present stage, arising from the low prevalence of high-grade specialized products. The expediency and possibilities of using microwave-drying vegetable powders for enriching fermented milk products for baby food with vitamins and dietary fiber are considered. A new fermented milk probiotic product with unique nutritional properties has been proposed, the optimal composition and features of its manufacture have been developed.

Pathogen-Specific Effects of Probiotics in Children with Acute Gastroenteritis Seeking Emergency Care: A Randomized Trial

Background It is unknown if probiotics exert pathogen-specific effects in children with diarrhea secondary to acute gastroenteritis. Methods Analysis of patient-level data from two multi-center randomized, placebo controlled, trials conducted in pediatric emergency departments in Canada and the U.S.. Participants were 3-48 months with >3 diarrheal episodes in the preceding 24 hours, and were symptomatic for <72 hours and <7 days in the Canadian and U.S. studies, respectively. Participants received either placebo or probiotic preparation (Canada-Lactobacillus rhamnosus R0011/L. helveticus R0052; U.S.-L. rhamnosus GG). The primary outcome was post-intervention moderate-to-severe disease [i.e. ≥9 on the Modified Vesikari Scale (MVS)]. Results Pathogens were identified in specimens from 59.3% (928/1565) children. No pathogen groups were less likely to experience an MVS ≥9 based on treatment allocation (test for interaction=0.35). No differences between groups were identified for adenovirus (aRR: 1.42; 95%CI: 0.62, 3.23), norovirus (aRR: 0.98; 95%CI: 0.56, 1.74), rotavirus (aRR: 0.86; 95%CI: 0.43, 1.71) or bacteria (aRR: 1.19; 95%CI: 0.41, 3.43). At pathogen-group and among individual pathogens there were no differences in diarrhea duration or the total number of diarrheal stools between treatment groups, regardless of intervention allocation or among probiotic sub-groups. Among adenovirus-infected children, those administered the Lactobacillus rhamnosus R0011/L. helveticus R0052 product experienced fewer diarrheal episodes (aRR: 0.65; 95%CI: 0.47 to 0.90). Conclusions Neither probiotic product resulted in less severe disease compared to placebo across a range of the most common etiologic pathogens. Clarifying the etiology of diarrhea among those without pathogens identified is warranted.

Production of a Potentially Probiotic Product for Animal Feed and Evaluation of Some of Its Probiotic Properties

Nowadays, probiotics have been proposed for substituting antibiotics in animal feed since the European Union banned the latter compounds in 2006 to avoid serious side effects on human health. Therefore, this work aimed to produce a probiotic product for use in animal feed by fed-batch fermentation of whey with a combination of kefir grains, AGK1, and the fermented whole milk used to activate these kefir grains. The probiotic culture obtained was characterized by high levels of biomass (8.03 g/L), total viability (3.6 × 108 CFU/mL) and antibacterial activity (28.26 Activity Units/mL). Some probiotic properties of the probiotic culture were investigated in vitro, including its survival at low pH values, under simulated gastrointestinal conditions, after freezing in skim milk at −20 °C, and in the commercial feed during storage at room temperature. The viable cells of lactic and acetic acid bacteria and yeasts exhibited higher tolerance to acidic pH and simulated gastrointestinal conditions when the cells were protected with skim milk and piglet feed, compared with washed cells. The results indicated the feasibility of producing a probiotic product at a low cost with a potential application in animal feed.

Virulence Factors and in-Host Selection on Phenotypes in Infectious Probiotic Yeast Isolates (Saccharomyces ‘boulardii’)

Saccharomyces yeast probiotics (S. ’boulardii’) have long been applied in the treatment of several gastrointestinal conditions. Despite their widespread use, they are rare opportunistic pathogens responsible for a high proportion of Saccharomyces mycosis cases. The potential virulence attributes of S. ’boulardii’ as well as its interactions with the human immune system have been studied, however, no information is available on how these yeasts may change due to in-host evolution. To fill this gap, we compared the general phenotypic characteristics, cell morphology, virulence factors, epithelial and immunological interactions, and pathogenicity of four probiotic product samples, two mycosis, and eight non-mycosis samples of S. ’boulardii’. We assessed the characteristics related to major steps of yeast infections. Mycosis and non-mycosis isolates both displayed novel characters when compared to the product isolates, but in the case of most virulence factors and in pathogenicity, differences were negligible or, surprisingly, the yeasts from products showed elevated levels. No isolates inflicted considerable damage to the epithelial model or bore the hallmarks of immune evasion. Our results show that strains in probiotic products possess characteristics that enable them to act as pathogens upon permissive conditions, and their entry into the bloodstream is not due to active mechanisms but depends on the host. Survival in the host is dependent on yeast phenotypic characteristics which may change in many ways once they start evolving in the host. These facts call attention to the shortcomings of virulence phenotyping in yeast research, and the need for a more thorough assessment of probiotic use.

Delivery of Metabolically Neuroactive Probiotics to the Human Gut

The human microbiome is a rich factory for metabolite production and emerging data has led to the concept that orally administered microbial strains can synthesize metabolites with neuroactive potential. Recent research from ex vivo and murine models suggests translational potential for microbes to regulate anxiety and depression through the gut-brain axis. However, so far, less emphasis has been placed on the selection of specific microbial strains known to produce the required key metabolites and the formulation in which microbial compositions are delivered to the gut. Here, we describe a double-capsule technology to deliver high numbers of metabolically active cells derived from the 24-strain probiotic product SH-DS01 to the gastrointestinal tract, including the small intestine, where immune responses and adsorption of metabolites into the bloodstream occur. Based on its genome sequence, Limosilactobacillus reuteri SD-LRE2-IT was predicted to have the genetic capacity to de novo produce a specific metabolite of interest to brain health, vitamin B12, which could be confirmed in vitro. Taken together, our data conceptualizes the importance of rationally defined microbial strain characterization based on genomics and metabolomics data, combined with carefully designed capsule technology for delivery of live cells and concomitant functionality in and beyond the gut ecosystem.

Oral probiotic and its delivery carriers to improve oral health: A review

In recent years, oral probiotics have been researched on their effectiveness in reducing and preventing oral diseases. Oral probiotics could be introduced into the oral cavity to keep the equilibrium of the microbiome. Hence, the delivery carrier for oral probiotics plays an important factor to ensure a high number of oral probiotics were delivered and released into the oral cavity. This review presents a brief overview of oral microbiota and the role of oral probiotics in reducing oral diseases. Moreover, important aspects of the oral probiotic product such as viability, adherence ability, health effects, safety, and delivery site were discussed. Besides that, the importance of utilizing indigenous oral probiotics was also emphasized. Oral probiotics are commonly found in the market in the form of chewing tablets, lozenges, and capsules. Hence, the oral probiotic carriers currently used in the market and research were reviewed. Furthermore, this review introduces new potential oral probiotic delivery carriers such as oral strip, bucco-adhesive gel, and mouthwash. Their effectiveness in delivering oral probiotics for oral health was also explored.

Production of a Potentially Probiotic Product for Animal Feed and Evaluation of Some of Its Probiotic Properties

Nowadays, probiotics has been proposed for substituting antibiotics in animal feed, since the European Union (EU) banned the latter compounds in 2006 to avoid their serious side effects on human health. Therefore, this work aimed to produce a probiotic product by fed-fermentation of whey with kefir grains for use in animal feed. The whey was fermented with a combination of kefir grains AGK1 and the free biomass present in whole milk fermented milk used to activate these kefir grains. The probiotic culture obtained was characterized with high levels of biomass, total viability and antibacterial activity. Some probiotic properties of the probiotic culture were investigated in vitro, including its survival at low pH values, under simulated gastrointestinal conditions, after freezing in skim milk at − 20 ºC, and in the commercial feed during storage at room temperature. The viable cells of lactic and acetic acid bacteria and yeasts exhibited higher tolerance to acidic pH and simulated gastrointestinal conditions when the cells were protected with skim milk and piglet feed, compared with washed cells. The results indicated the feasibility of producing a probiotic product at a low cost with a potential application in animal feed.