Human milk: microbiota composition and impact of processing on probiotic properties

Preterm birth complications represent a leading cause of mortality in children under five years old. To mitigate these complications, strategies such as the use of donor human milk, fortification with human milk fortifiers (protein supplements), and probiotic administration are employed. This thesis aimed to thoroughly understand the microbiota composition of preterm human milk and the influence of various factors on specific probiotics' properties. The study began with analyzing human milk microbiota from healthy Argentinian women across different gestational ages, focusing on changes in microbiota from birth to term equivalent age. It was found that term milk exhibited greater microbial diversity than preterm milk, and by term equivalent age, the microbiota composition of human milk closely resembled that of term milk, suggesting breastfeeding's role in promoting microbial maturity. The research further assessed the stability of probiotics, specifically Lacticaseibacillus rhamnosus GG and Bifidobacterium animalis ssp. lactis Bb12, during cold storage of human milk. Results indicated that these probiotics maintained their viability for up to 72 hours. Additionally, this thesis examined the effect of liquid or powdered human milk fortifiers on the adhesion properties of these probiotics in donor human milk, finding that fortification did not adversely affect probiotic adhesion. The final study of the thesis explored the role of polyamines, in modulating the adhesion of probiotics and pathogens with intestinal mucus in infants. The presence of the polyamine spermidine significantly enhanced the adhesion of Bb12 in infant mucus under six months, whereas the polyamine spermine reduced the adhesion of Cronobacter sakazakii. In conclusion, the thesis showed that the microbiota profiles of human milk significantly vary with gestational age. A key finding is that the microbiota composition of preterm human milk undergoes a transformation, eventually resembling that of term milk as the infant reaches term equivalent age. It was also found that probiotic adhesion properties are unaffected by cold storage or the fortification of donor human milk. However, the influence of polyamines on bacterial adhesion to mucus varies based on the bacterial strain and the mucus donor's age employed. These insights should be considered for developing optimized gut colonization strategies for infants, particularly for those born preterm.Äidinmaito: mikrobiston koostumus ja prosessoinnin vaikutus sen probioottisiin ominaisuuksiin Ennenaikaisen synnytyksen komplikaatiot ovat yleisin alle viisivuotiaiden lasten kuolinsyy. Luovutettua äidinmaitoa, äidinmaidon täydentämistä proteiinilisällä sekä probioottien antamista käytetään näiden komplikaatioiden ehkäisemiseksi. Tämän opinnäytetyön tavoitteena oli selvittää ennenaikaisesti synnyttäneiden äitien rintamaidon mikrobistokoostumusta ja eri tekijöiden vaikutusta tiettyjen probioottien ominaisuuksiin. Tutkimuksessa analysoitiin terveiden argentiinalaisten naisten äidinmaidon mikrobikoostumusta keskittyen mikrobiston muutoksiin ennenaikaisesta synnytyksestä laskettuun aikaan. Huomattiin, että täysiaikaisesti synnyttäneiden naisten äidinmaidon mikrobiston monimuotoisuus oli suurempaa kuin ennenaikaisesti synnyttäille naisilla. Laskettuna aikana ennenaikasesti synnyttäneiden äidinmaidon mikrobikoostumus muistutti läheisesti täysiaikaisen maidon mikrobistoa, mikä viittaa siihen, että imetyksellä on tärkeä rooli mikrobiston kypsymisen edistämisessä. Tutkimuksessa, jossa arvioitiin Lacticaseibacillus rhamnosus GG:n ja Bifidobacterium animalis ssp. lactis Bb12 -probioottien stabiiliutta äidinmaidossa kylmävarastoinnin aikana, nämä probiootit säilyttivät elinkykynsä jopa 72 tuntia. Lisäksi havaittiin, että äidinmaidon vahvistaminen nestemäisellä tai jauhemaisella vahvikkeella ei vaikuttanut haitallisesti probioottien tarttumisominaisuuksiin luovutetussa äidinmaidossa. Opinnäytetyön viimeisessä tutkimuksessa selvitettiin polyamiinien roolia bakteerien tarttumisen säätelyssä imäveisen suolistolimassa. Polyamiini spermidiinin läsnäolo paransi merkittävästi Bb12:n kiinnittymistä alle kuuden kuukauden ikäisen imeväisten limaan, kun taas spermiini vähensi Cronobacter sakazakii-bakteerin tarttumista. Opinnäytetyö osoitti, että äidinmaidon mikrobistoprofiilit vaihtelevat merkittävästi raskauden keston mukaan. Ennenaikaisen synnytyksen jälkeen äidinmaidon mikrobistokoostumus muuttuu ja alkaa lopulta muistuttaa täysiaikaisen synnytyksen äidinmaidon mikrobistokoostumusta vauvan saavuttaessa taysiaikaista vastaavan iän. Lisäksi havaittiin, että kylmävarastointi tai luovutetun äidinmaidon vahvistaminen eivät vaikuttaneet probioottien tarttumisominaisuuksiin. Polyamiinien vaikutus bakteerien tarttumiseen limaan vaihtelee bakteerikannan ja limanluovuttajan iän mukaan. Tulokset auttavat kehittämään optimoituja suoliston kolonisaatiostrategioita vastasyntyneille ja erityisesti keskosille

Factors influencinlag the microbial composition of human milk

Aside from nutritional components, human milk is rich in microorganisms. Through breastfeeding these microorganisms are introduced to the infant gut where they may transiently or persistently colonize it. Therefore, the human milk microbiota may be an important factor which shapes the infant gut microbiota further influencing infant health and disease. In the current review we aim to give a brief updated insight into the putative origin of the human milk microbiota, its constituents and the possible factors that shape it. Understanding the factors that determine the human milk microbiota composition and function will aid developing optimal postnatal feeding and intervention strategies to reduce the risk of communicable and noncommunicable diseases. (C) 2021 The Authors. Published by Elsevier Inc.Peer reviewe

Postbiotics against pathogens commonly involved in pediatric infectious diseases

The Sustainable Development goals for 2020 included reducing all causes associated with infant and perinatal mortality in their priorities. The use of compounds with bioactive properties has been proposed as a therapeutic strategy due to their stimulating effect on the host’s immune system. Additionally, biotherapeutic products such as postbiotics, tentatively defined as compounds produced during a fermentation process that support health and well-being, promote intestinal barrier integrity without posing considerable risks to children’s health. Although this is a concept in development, there are increasing studies in the field of nutrition, chemistry, and health that aim to understand how postbiotics can help prevent different types of infections in priority populations such as minors under the age of five. The present review aims to describe the main mechanisms of action of postbiotics. In addition, it presents the available current evidence regarding the effects of postbiotics against pathogens commonly involved in pediatric infections. Postbiotics may constitute a safe alternative capable of modulating the cellular response and stimulating the host’s humoral response

Postbiotics for preventing and treating common infectious diseases in children: A systematic review

Postbiotics have recently been tentatively defined as bioactive compounds produced during a fermentation process (including microbial cells, cell constituents and metabolites) that supports health and/or wellbeing. Postbiotics are currently available in some infant formulas and fermented foods. We systematically reviewed evidence on postbiotics for preventing and treating common infectious diseases among children younger than 5 years. The PubMed, Embase, SpringerLink, and ScienceDirect databases were searched up to March 2019 for randomized controlled trials (RCTs) comparing postbiotics with placebo or no intervention. Seven RCTs involving 1740 children met the inclusion criteria. For therapeutic trials, supplementation with heat-killed Lactobacillus acidophilus LB reduced the duration of diarrhea (4 RCTs, n = 224, mean difference, MD, −20.31 h, 95% CI −27.06 to −13.57). For preventive trials, the pooled results from two RCTs (n = 537) showed that heat‐inactivated L. paracasei CBA L74 versus placebo reduced the risk of diarrhea (relative risk, RR, 0.51, 95% CI 0.37–0.71), pharyngitis (RR 0.31, 95% CI 0.12–0.83) and laryngitis (RR 0.44, 95% CI 0.29–0.67). There is limited evidence to recommend the use of specific postbiotics for treating pediatric diarrhea and preventing common infectious diseases among children. Further studies are necessary to determine the effects of different postbiotic

Postbiotics for Preventing and Treating Common Infectious Diseases in Children: A Systematic Review

Postbiotics have recently been tentatively defined as bioactive compounds produced during a fermentation process (including microbial cells, cell constituents and metabolites) that supports health and/or wellbeing. Postbiotics are currently available in some infant formulas and fermented foods. We systematically reviewed evidence on postbiotics for preventing and treating common infectious diseases among children younger than 5 years. The PubMed, Embase, SpringerLink, and ScienceDirect databases were searched up to March 2019 for randomized controlled trials (RCTs) comparing postbiotics with placebo or no intervention. Seven RCTs involving 1740 children met the inclusion criteria. For therapeutic trials, supplementation with heat-killed Lactobacillus acidophilus LB reduced the duration of diarrhea (4 RCTs, n = 224, mean difference, MD, −20.31 h, 95% CI −27.06 to −13.57). For preventive trials, the pooled results from two RCTs (n = 537) showed that heat-inactivated L. paracasei CBA L74 versus placebo reduced the risk of diarrhea (relative risk, RR, 0.51, 95% CI 0.37-0.71), pharyngitis (RR 0.31, 95% CI 0.12-0.83) and laryngitis (RR 0.44, 95% CI 0.29-0.67). There is limited evidence to recommend the use of specific postbiotics for treating pediatric diarrhea and preventing common infectious diseases among children. Further studies are necessary to determine the effects of different postbiotics.</p

Postbiotics against Pathogens Commonly Involved in Pediatric Infectious Diseases

The Sustainable Development goals for 2020 included reducing all causes associated with infant and perinatal mortality in their priorities. The use of compounds with bioactive properties has been proposed as a therapeutic strategy due to their stimulating effect on the host’s immune system. Additionally, biotherapeutic products such as postbiotics, tentatively defined as compounds produced during a fermentation process that support health and well-being, promote intestinal barrier integrity without posing considerable risks to children’s health. Although this is a concept in development, there are increasing studies in the field of nutrition, chemistry, and health that aim to understand how postbiotics can help prevent different types of infections in priority populations such as minors under the age of five. The present review aims to describe the main mechanisms of action of postbiotics. In addition, it presents the available current evidence regarding the effects of postbiotics against pathogens commonly involved in pediatric infections. Postbiotics may constitute a safe alternative capable of modulating the cellular response and stimulating the host’s humoral response.</p

Exogenous Polyamines Influence In Vitro Microbial Adhesion to Human Mucus according to the Age of Mucus Donor

Adhesion to intestinal mucus is the first step for microbiota colonization in early life. Polyamines are polycations with important physiological functions in both procaryotic and eucaryotic cells. However, their role in intestinal mucus adhesion is not known. The objective of the present study was to evaluate whether exogenous polyamines (putrescine, spermidine, spermine, and their combination) would alter the adhesive properties of Lacticaseibacillus rhamnosus GG (LGG), Bifidobacterium animalis subs. lactis Bb12, Cronobacter sakazakii, and Escherichia coli. Human intestinal mucus was isolated from healthy infants (0–6-month-old and 6–12-month-old) and healthy adults (25–52 years old). Spermidine significantly increased Bb12 adhesion (p </p

The Effect of Donor Human Milk Fortification on The Adhesion of Probiotics In Vitro

Preterm delivery complications are the primary cause of death among children under the age of five. Preventive strategies include the use of pasteurized donor human milk (DHM), its fortification with human milk fortifiers (protein supplements), and supplementation with probiotics. Our aim was to examine the impact of DHM and fortified DHM (FDHM) on the mucus adhesion properties of two widely used probiotics. The study covered two forms of human milk fortifier, liquid and powdered, with or without probiotics and storage at 4 °C for 24 h. To test the adhesion properties of the probiotic strains, DHM+probiotics and FDHM+probiotics were prepared and added to immobilized mucus isolated from the stool of healthy Finnish infants. The probiotic adhesion was then measured by liquid scintillation. Our results suggest that addition of liquid or powdered human milk fortifier in donor human milk had no impact on probiotic adhesion. In addition, given the increased adhesion of probiotics suspended in buffer, other matrices should be further studied. These factors need to be considered when designing future intervention strategies using probiotics in preterm infants.</p

Bifidobacterium animalis subsp lactis HN019 presents antimicrobial potential against periodontopathogens and modulates the immunological response of oral mucosa in periodontitis patients

Objective To evaluate the effects of Bifidobacterium animalis subsp. lactis HN019 (HN019) on clinical periodontal parameters (plaque accumulation and gingival bleeding), on immunocompetence of gingival tissues [expression of beta-defensin (BD)-3, toll-like receptor 4 (TLR4), cluster of differentiation(CD)-57 and CD-4], and on immunological properties of saliva (IgA levels) in non-surgical periodontal therapy in generalized chronic periodontitis (GCP) patients. Adhesion to buccal epithelial cells (BEC) and the antimicrobial properties of HN019 were also investigated. Materials and methods Thirty patients were recruited and monitored clinically at baseline (before scaling and root planing-SRP) and after 30 and 90 days. Patients were randomly assigned to Test (SRP +Probiotic, n = 15) or Control (SRP+Placebo, n = 15) group. Probiotic lozenges were used for 30 days. Gingival tissues and saliva were immunologically analyzed. The adhesion of HN019 with or without Porphyromonas gingivalis in BEC and its antimicrobial properties were investigated in in vitro assays. Data were statistically analyzed (p<0.05). Results Test group presented lower plaque index (30 days) and lower marginal gingival bleeding (90 days) when compared with Control group. Higher BD-3, TLR4 and CD-4 expressions were observed in gingival tissues in Test group than in Control group. HN019 reduced the adhesion of P. gingivalis to BEC and showed antimicrobial potential against periodontopathogens. Conclusion Immunological and antimicrobial properties of B. lactis HN019 make it a potential probiotic to be used in non-surgical periodontal therapy of patients with GCP.Fil: Invernici, Marcos M.. Universidade de Sao Paulo; BrasilFil: Furlaneto, Flávia A. C.. Universidade de Sao Paulo; BrasilFil: Salvador, Sérgio L. Universidade de Sao Paulo; BrasilFil: Ouwehand, Arthur C.. Dupont, Nutrition and Health; FinlandiaFil: Salminen, Seppo. University of Turku. Functional Foods Forum; FinlandiaFil: Mantziari, Anastasia. University of Turku. Functional Foods Forum; FinlandiaFil: Vinderola, Celso Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Ervolino, Edilson. São Paulo State University. Division of Histology, Department of Basic Sciences, Dental School of Araçatuba; BrasilFil: Santana, Sandro Isaías. Universidade de Sao Paulo; BrasilFil: Silva, Pedro Henrique Felix. Universidade de Sao Paulo; BrasilFil: Messora, Michel R.. Universidade de Sao Paulo; Brasi

Bifidobacterium animalis subsp lactis HN019 presents antimicrobial potential against periodontopathogens and modulates the immunological response of oral mucosa in periodontitis patients

Objective To evaluate the effects of Bifidobacterium animalis subsp. lactis HN019 (HN019) on clinical periodontal parameters (plaque accumulation and gingival bleeding), on immunocompetence of gingival tissues [expression of beta-defensin (BD)-3, toll-like receptor 4 (TLR4), cluster of differentiation(CD)-57 and CD-4], and on immunological properties of saliva (IgA levels) in non-surgical periodontal therapy in generalized chronic periodontitis (GCP) patients. Adhesion to buccal epithelial cells (BEC) and the antimicrobial properties of HN019 were also investigated. Materials and methods Thirty patients were recruited and monitored clinically at baseline (before scaling and root planing—SRP) and after 30 and 90 days. Patients were randomly assigned to Test (SRP+Probiotic, n = 15) or Control (SRP+Placebo, n = 15) group. Probiotic lozenges were used for 30 days. Gingival tissues and saliva were immunologically analyzed. The adhesion of HN019 with or without Porphyromonas gingivalis in BEC and its antimicrobial properties were investigated in in vitro assays. Data were statistically analyzed (p</p