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1.
Nutr. hosp ; 38(1): 152-160, ene.-feb. 2021. tab, graf
Artigo em Inglês | IBECS | ID: ibc-198852

RESUMO

INTRODUCTION: breast milk (MH) contains nutrients and bioactive compounds for child development, including probiotic bacteria, which contribute to intestinal maturation. This benefit accompanies the individual until adulthood. There are new methods such as spray drying that give this compound a good conservation without loss of microbiota. OBJECTIVE: the aim of this study was to analyze the viability of lactic acid bacteria isolated from human milk with probiotic potential after the spray drying process, as well as to evaluate the possible adhesion in the colon of mice of the Balb/C strain after feeding them powdered human milk and a commercial formula milk. METHOD: we isolated and identified the presence of lactic acid bacteria with possible probiotic potential in powdered human milk using the MALDI-TOF MS technique. Powdered human milk and a commercial formula milk were fed to mice of the Bald/C strain for 14 weeks. Glucose level and weight were measured in the mice. The feces were collected to verify the presence of lactic bacteria. The mice were sacrificed and their intestines were weighed, isolating the lactic acid bacteria both from the intestines and from the feces. The strains isolated from mice fed human milk were evaluated for their probiotic potential, analyzing their ability to inhibit pathogens, resistance to pH, temperature, adhesion, and hydrophobicity. RESULTS: the presence of Lactobacillus fermentum LH01, Lactobacillus rhamnosus LH02, Lactobacullis reuteri LH03, and Lactobacillus plantarum LH05 in powdered human milk was identified. All strains showed a possible probiotic profile due to the ability of bacteria to resist low pH, bile salts, and exposure to gastric enzymes, as well as their hydrophobicity and self-aggregation capacity, and their failure to show hemagglutination or hemolysis activity in a culture medium rich in erythrocytes. We observed that the consumption of powdered human milk prevented weight gain and constipation in mice. CONCLUSIONS: after spray drying, strains with possible probiotic potential may be preserved in human milk. The consumption of powdered human milk with probiotic bacteria prevents constipation and weight gain in mice, when compared to those fed a commercial formula milk


INTRODUCCIÓN: la leche materna (HM) contiene los nutrientes y compuestos bioactivos necesarios para el desarrollo infantil, incluidas bacterias probióticas, que contribuyen a la maduración intestinal. OBJETIVO: el objetivo de este estudio fue analizar la viabilidad de las bacterias acidolácticas aisladas de la leche humana con potencial probiótico, después del proceso de secado, así como evaluar su posible adhesión en el colón de ratones (BAlb/C) alimentados con leche humana en polvo y leche de una fórmula comercial. MÉTODO: se aislaron e identificaron mediante la técnica de Maldi-Tof-MS las bacterias acidolácticas con posible potencial probiótico en la leche humana en polvo. Se alimentó con leche humana en polvo y leche de una fórmula comercial a ratones de la cepa Bald/C durante 14 semanas. Se midieron el nivel de glucosa y el peso. Las heces se recolectaron para verificar la presencia de bacterias lácticas. Los ratones se sacrificaron y se pesaron los intestinos, aislando las bacterias lácticas tanto de los intestinos como de las heces. En las cepas aisladas de la leche humana se evaluó el potencial probiótico analizando su capacidad para inhibir patógenos, resistir distintos pH y temperaturas, adherirse y mostrar hidrofobicidad. RESULTADOS: se identificó la presencia de Lactobacillus fermentum LH01, Lactobacillus rhamnosus LH02, Lactobacullis reuteri LH03 y L. plantarum LH05 en la leche humana en polvo. Todas las cepas mostraron resistencia a los pH bajos, a las sales biliares y a la exposición a enzimas gástricas, así como una buena hidrofobicidad y capacidad de autoagregación. Además, no presentaron actividad de hemaglutinación o hemólisis en un medio de cultivo rico en eritrocitos. Observamos que el consumo de leche humana en polvo evita en los ratones el aumento de peso y el estreñimiento. CONCLUSIONES: después del secado por aspersión, las cepas con posible potencial probiótico pueden conservarse en la leche materna. El consumo de leche humana en polvo con bacterias probióticas evita el estreñimiento y el aumento de peso en los ratones, en comparación con los alimentados con leche de una formula comercial


Assuntos
Animais , Camundongos , Leite Humano/microbiologia , Probióticos , Lactobacillus fermentum/isolamento & purificação , Lactobacillus plantarum/isolamento & purificação , Obesidade/prevenção & controle , Constipação Intestinal/prevenção & controle , Lactobacillus reuteri/isolamento & purificação , Lactobacillus fermentum/fisiologia , Lactobacillus plantarum/fisiologia , Constipação Intestinal/veterinária , Obesidade/veterinária
2.
Microbiome ; 9(1): 41, 2021 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-33568231

RESUMO

BACKGROUND: Quality control including assessment of batch variabilities and confirmation of repeatability and reproducibility are integral component of high throughput omics studies including microbiome research. Batch effects can mask true biological results and/or result in irreproducible conclusions and interpretations. Low biomass samples in microbiome research are prone to reagent contamination; yet, quality control procedures for low biomass samples in large-scale microbiome studies are not well established. RESULTS: In this study, we have proposed a framework for an in-depth step-by-step approach to address this gap. The framework consists of three independent stages: (1) verification of sequencing accuracy by assessing technical repeatability and reproducibility of the results using mock communities and biological controls; (2) contaminant removal and batch variability correction by applying a two-tier strategy using statistical algorithms (e.g. decontam) followed by comparison of the data structure between batches; and (3) corroborating the repeatability and reproducibility of microbiome composition and downstream statistical analysis. Using this approach on the milk microbiota data from the CHILD Cohort generated in two batches (extracted and sequenced in 2016 and 2019), we were able to identify potential reagent contaminants that were missed with standard algorithms and substantially reduce contaminant-induced batch variability. Additionally, we confirmed the repeatability and reproducibility of our results in each batch before merging them for downstream analysis. CONCLUSION: This study provides important insight to advance quality control efforts in low biomass microbiome research. Within-study quality control that takes advantage of the data structure (i.e. differential prevalence of contaminants between batches) would enhance the overall reliability and reproducibility of research in this field. Video abstract.


Assuntos
Microbiota , Leite Humano/microbiologia , Adulto , Animais , Pré-Escolar , Feminino , Humanos , Lactente , Microbiota/genética , Reprodutibilidade dos Testes
3.
J Dairy Sci ; 104(2): 1518-1523, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33358168

RESUMO

The oligosaccharide 2'-fucosyllactose (2'FL) in human breast milk selectively promotes the proliferation of bifidobacteria. One hundred fifty-one Bifidobacterium strains were evaluated for their capacity to utilize 2'FL based on the combination of phenotype and genotype association analysis. Through genotype analysis, 37 strains were predicted to have the ability to use 2'FL, including Bifidobacteriumbifidum, Bifidobacteriumbreve, Bifidobacteriumlongum ssp. longum, Bifidobacteriumlongum ssp. infantis, and Bifidobacteriumdentium, whereas Bifidobacteriumadolescentis, Bifidobacteriumanimalis, Bifidobacteriumpseudocatenulatum, and Bifidobacteriumangulatum could not use 2'FL. For in vitro utilization, there were noteworthy differences for 2'FL usage among different species, which were 100% consistent with genotype prediction. The results indicated that 2'FL utilization ability differed even within the same species, and Bifidobacterium followed the currently well-known pathway to utilize 2'FL, which could provide guidance to develop personalized prebiotics for different bifidobacteria via gene-trait matching analysis.


Assuntos
Bifidobacterium/metabolismo , Leite Humano/microbiologia , Trissacarídeos/metabolismo , Animais , Bifidobacterium/genética , Estudos de Associação Genética , Humanos , Oligossacarídeos/metabolismo , Prebióticos
4.
BMC Med ; 18(1): 392, 2020 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-33317529

RESUMO

BACKGROUND: The proportion of infections among young children that are antimicrobial-resistant is increasing across the globe. Newborns may be colonized with enteric antimicrobial-resistant pathogens early in life, which is a risk factor for infection-related morbidity and mortality. Breastfeeding is actively promoted worldwide for its beneficial impacts on newborn health and gut health. However, the role of breastfeeding and human milk components in mitigating young children's carriage of antimicrobial-resistant pathogens and antibiotic resistance genes has not been comprehensively explored. MAIN BODY: Here, we review how the act of breastfeeding, early breastfeeding, and/or human milk components, such as the milk microbiota, secretory IgA, human milk oligosaccharides, antimicrobial peptides, and microRNA -bearing extracellular vesicles, could play a role in preventing the establishment of antimicrobial-resistant pathogens in young children's developing gut microbiomes. We describe findings from recent human studies that support this concept. CONCLUSION: Given the projected rise in global morbidity and mortality that will stem from antimicrobial-resistant infections, identifying behavioral or nutritional interventions that could decrease children's susceptibility to colonization with antimicrobial-resistant pathogens may be one strategy for protecting their health. We suggest that breastfeeding and human milk supplements deserve greater attention as potential preventive measures in the global effort to combat antimicrobial resistance, particularly in low- and middle-income settings.


Assuntos
Aleitamento Materno/métodos , Farmacorresistência Bacteriana/fisiologia , Microbioma Gastrointestinal/imunologia , Leite Humano/microbiologia , Feminino , Humanos , Recém-Nascido , Masculino
5.
PLoS One ; 15(5): e0233284, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32469943

RESUMO

Human milk provides complete nutrition for infants and at the same time promotes the growth of specific bacteria in the infant gastrointestinal tract. Breastfeeding can often be discontinued due to mastitis which is an inflammation of the breast tissue. We isolated 18 Staphylococcus aureus strains from milk donated by healthy (n = 6), subclinical (n = 6), and mastitic (n = 6) mothers, two strains of which were VISA (Vancomycin Intermediate S. aureus). All tested strains (n = 12) were able to form biofilms. We then examined the impact of nisin A and vancomycin alone and in combination on biofilm formation and eradication of selected strains (n = 8). We observed strain-specific responses, with the combinatorial treatment at 1/4X MIC (for both singularly) significantly inhibiting biofilm formation for seven out of eight strains when compared with nisin A or vancomycin alone. None of the selected treatments were able to eradicate pre-formed biofilms. Finally, we selected two strains, namely a VISA (APC3814H) and a strong biofilm former (APC3912CM) and used confocal microscopy to evaluate the effects of the antimicrobial agents at 1X MIC on biofilm inhibition and eradication. All treatments inhibited biofilm formation of APC3814H but were ineffective in eradicating a pre-formed biofilm. Single treatments at 1X MIC against APC3912CM cells did not prevent biofilm formation whereas combination treatment caused increased death of APC3912CM cells. Finally, the combination treatment reduced the thickness of the pre-formed APC3912CM biofilm as compared with the single treatments.


Assuntos
Biofilmes/efeitos dos fármacos , Mastite/tratamento farmacológico , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Leite Humano/efeitos dos fármacos , Nisina/uso terapêutico , Infecções Estafilocócicas/tratamento farmacológico , Vancomicina/uso terapêutico , Antibacterianos/uso terapêutico , Biofilmes/crescimento & desenvolvimento , Quimioterapia Combinada , Feminino , Humanos , Mastite/microbiologia , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Testes de Sensibilidade Microbiana , Leite Humano/microbiologia , Infecções Estafilocócicas/microbiologia
6.
Food Microbiol ; 89: 103450, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32138997

RESUMO

This study was conducted to address the dearth in works that simultaneously compare the growth and inactivation behaviors of selected pathogens in different milk products. In worst-case scenarios where hygienic practices are absent and heavy microbiological contaminations occur, Salmonella enterica, Escherichia coli O157:H7, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus multiplied in all samples at room temperature (27 °C). Most organisms readily proliferated with growth lag (tlag) values ranging from 0.00 to 5.95 h. Growth rates (KG) ranged from 0.16 to 0.67 log CFU/h. Sanitary risk times (SRTs) for a 1-log population increase ranged from 1.85 to 6.27 h, while 3.69-12.55 h were the SRTs determined for 2-log population increase. Final populations (Popfin) ranged from 7.11 to 9.36 log CFU/mL. Inactivation in heavily contaminated milk during Holder pasteurization revealed biphasic inactivation behavior with total log reduction (TLR) after exposure to 62.5 °C for 30 min ranging from 1.91 (90.8%) to 6.00 (99.9999%). These results emphasize the importance food safety systems in the handling of milk and milk products during manufacture and preparation.


Assuntos
Bactérias/crescimento & desenvolvimento , Inocuidade dos Alimentos , Fórmulas Infantis/microbiologia , Leite Humano/microbiologia , Leite/microbiologia , Animais , Bovinos , Escherichia coli O157/crescimento & desenvolvimento , Feminino , Microbiologia de Alimentos , Humanos , Lactente , Listeria monocytogenes/crescimento & desenvolvimento , Pasteurização , Salmonella enterica/crescimento & desenvolvimento , Staphylococcus aureus/crescimento & desenvolvimento , Temperatura
7.
Arch Microbiol ; 202(6): 1425-1438, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32185412

RESUMO

The early bifidobacterial colonization and development of infant gut is considered crucial for the immediate and lifelong health of human host. This study longitudinally analyzed and characterized fecal bifidobacterial profiles in association with feeding regimens observed in six infants during 5 months after birth. The dominant fecal microbiota of bifidobacteria, lactobacilli/enterococci, clostridia, bacteroides and eubacteria were specifically enumerated using fluorescent in situ hybridization (FISH) technique. Breastfeeding exhibited close association with the predomination of bifidobacteria with the highest relative abundance of 32-70% detected in both infants with exclusive breastfeeding. The nested PCR-DGGE technique revealed high diversity existing within a bifidobacterial species with multiple strain variants of B. bifidum, B. longum, B. breve and B. dentium continuously detected in feces of exclusively breast- and combination-fed infants over the period of 5 months. Contrarily, B. breve, B. adolescentis, B. dentium, B. bifidum, B. faecale, B. kashiwanohense and B. lactis detected in all exclusively formula-fed infants seem to be transient species. The persisting strains seem to derive primarily from maternal breastmilk as demonstrated by PCR-DGGE profiles of human milk and feces from three mother-infant pairs. The results suggested the pivotal role of breastfeeding regimen in supporting colonization and succession of bifidobacteria in infant gut.


Assuntos
Bifidobacterium/classificação , Bifidobacterium/isolamento & purificação , Microbioma Gastrointestinal/fisiologia , Fórmulas Infantis/microbiologia , Leite Humano/microbiologia , Bactérias Anaeróbias , Bifidobacterium/genética , Aleitamento Materno , Fezes/microbiologia , Humanos , Hibridização in Situ Fluorescente , Lactente , Recém-Nascido , Reação em Cadeia da Polimerase , Tailândia
8.
Sensors (Basel) ; 20(4)2020 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-32093069

RESUMO

There are no existing affordable diagnostics for sensitive, rapid, and on-site detection of pathogens in milk. To this end, an on-site colorimetric-based sustainable assay has been developed and optimized using an L16 (54) Taguchi design to obtain results in hours without PCR amplification. To determine the level of Escherichia coli (E. coli) contamination, after induction with 150 µL of breast milk, the B-Per bacterial protein extraction kit was added to a solution containing an alginate-based microcapsule assay. Within this 3 mm spherical novel sensor design, X-Gal (5-Bromo-4-Chloro-3-Indolyl ß-d-Galactopyranoside) was entrapped at a concentration of 2 mg/mL. The outward diffusing X-Gal was cleaved by ß-galactosidase from E. coli and dimerized in the solution to yield a blue color after incubation at 40 °C. Color intensity was correlated with the level of E. coli contamination using a categorical scale. After an 8 h incubation period, a continuous imaging scale based on intensity normalization was used to determine a binary lower limit of detection (LOD), which corresponded to 102 colony forming unit per mL (CFU/mL) and above. The cost of the overall assay was estimated to be $0.81 per sample, well under the $3 benchmark for state-of-the-art immune-based test kits for pathogen detection in biofluids. Considering the reported binary LOD cutoff of 102 CFU/mL and above, this proposed hydrogel-based assay is suited to meet global requirements for screening breast milk or milk for pathogenic organisms of 104 CFU/mL, with a percentage of false positives to be determined in future efforts.


Assuntos
Alginatos/química , Técnicas Biossensoriais/métodos , Escherichia coli/isolamento & purificação , Leite Humano/microbiologia , Catálise , Humanos , Limite de Detecção , Padrões de Referência , Razão Sinal-Ruído
9.
Nutrients ; 12(2)2020 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-32069822

RESUMO

Cardiorespiratory function is not only the foremost determinant of life after premature birth, but also a major factor of long-term outcomes. However, the path from placental disconnection to nutritional autonomy is enduring and challenging for the preterm infant and, at each step, will have profound influences on respiratory physiology and disease. Fluid and energy intake, specific nutrients such as amino-acids, lipids and vitamins, and their ways of administration -parenteral or enteral-have direct implications on lung tissue composition and cellular functions, thus affect lung development and homeostasis and contributing to acute and chronic respiratory disorders. In addition, metabolomic signatures have recently emerged as biomarkers of bronchopulmonary dysplasia and other neonatal diseases, suggesting a profound implication of specific metabolites such as amino-acids, acylcarnitine and fatty acids in lung injury and repair, inflammation and immune modulation. Recent advances have highlighted the profound influence of the microbiome on many short- and long-term outcomes in the preterm infant. Lung and intestinal microbiomes are deeply intricated, and nutrition plays a prominent role in their establishment and regulation. There is an emerging evidence that human milk prevents bronchopulmonary dysplasia in premature infants, potentially through microbiome composition and/or inflammation modulation. Restoring antibiotic therapy-mediated microbiome disruption is another potentially beneficial action of human milk, which can be in part emulated by pre- and probiotics and supplements. This review will explore the many facets of the gut-lung axis and its pathophysiology in acute and chronic respiratory disorders of the prematurely born infant, and explore established and innovative nutritional approaches for prevention and treatment.


Assuntos
Doenças do Prematuro/metabolismo , Pneumopatias/fisiopatologia , Microbiota/fisiologia , Nutrientes/metabolismo , Nascimento Prematuro/fisiopatologia , Feminino , Microbioma Gastrointestinal/fisiologia , Humanos , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido , Recém-Nascido Prematuro/crescimento & desenvolvimento , Doenças do Prematuro/etiologia , Doenças do Prematuro/microbiologia , Pulmão/crescimento & desenvolvimento , Pulmão/microbiologia , Pneumopatias/etiologia , Pneumopatias/microbiologia , Masculino , Leite Humano/microbiologia , Placenta/microbiologia , Gravidez , Nascimento Prematuro/microbiologia
10.
Dig Dis Sci ; 65(3): 723-740, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32060812

RESUMO

This review summarizes the key results of recently published studies on the effects of dietary change and nutritional intervention on the human microbiome from around the world, focusing on the USA, Canada, Europe, Asia, and Africa. It first explores mechanisms that might explain the ability of fiber-rich foods to suppress the incidence and mortality from westernized diseases, notably cancers of the colon, breast, liver, cardiovascular, infectious, and respiratory diseases, diabetes, and obesity (O'Keefe in Lancet Gastroenterol Hepatol 4(12):984-996, 2019; Am J Clin Nutr 110:265-266, 2019). It summarizes studies from Africa which suggest that disturbance of the colonic microbiome may exacerbate chronic malnutrition and growth failure in impoverished communities and highlights the importance of breast feeding. The American section discusses the role of the microbiome in the swelling population of patients with obesity and type 2 diabetes and examines the effects of race, ethnicity, geography, and climate on microbial diversity and metabolism. The studies from Europe and Asia extoll the benefits of whole foods and plant-based diets. The Asian studies examine the worrying changes from low-fat, high-carbohydrate diets to high-fat, low-carbohydrate ones and the increasing appearance of westernized diseases as in Africa and documents the ability of high-fiber traditional Chinese diets to reverse type 2 diabetes and control weight loss. In conclusion, most of the studies reviewed demonstrate clear changes in microbe abundances and in the production of fermentation products, such as short-chain fatty acids and phytochemicals following dietary change, but the significance of the microbiota changes to human health, with the possible exception of the stimulation of butyrogenic taxa by fiber-rich foods, is generally implied and not measured. Further studies are needed to determine how these changes in microbiota composition and metabolism can improve our health and be used to prevent and treat disease.


Assuntos
Dieta , Fibras na Dieta/microbiologia , Microbioma Gastrointestinal/fisiologia , Internacionalidade , Leite Humano/microbiologia , Dieta/tendências , Dieta Ocidental/efeitos adversos , Humanos , Leite Humano/fisiologia
11.
Dig Dis Sci ; 65(3): 706-722, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32002758

RESUMO

Diet is a key regulator of microbiome structure and function across the lifespan. Microbial colonization in the first year of life has been actively researched; however, studies during childhood are sparse. Herein, the impact of dietary intake and pre- and probiotic interventions on microbiome composition of healthy infants and children from birth to adolescence is discussed. The microbiome of breastfed infants has lower microbial diversity and richness, higher Proteobacteria, and lower Bacteroidetes and Firmicutes than those formula-fed. As children consume more complex diets, associations between dietary patterns and the microbiota emerge. Like adults, the microbiota of children consuming a Western-style diet is associated with greater Bacteroidaceae and Ruminococcaceae and lower Prevotellaceae. Dietary fibers and pre- or/and probiotics have been tested to modulate the gut microbiota in early life. Human milk oligosaccharides and prebiotics added to infant formula are bifidogenic and decrease pathogens. In children, prebiotics, such as inulin, increase Bifidobacterium abundance and dietary fibers reduce fecal pH and increase alpha diversity and calcium absorption. Probiotics have been administered to the mother during pregnancy and breastfeeding or directly to the infant/child. Findings on maternal probiotic administration on bacterial taxa are inconsistent. When given directly to the infant/child, some changes in individual taxa are observed, but rarely is overall alpha or beta diversity affected. Cesarean-delivered infants appear to benefit to a greater degree than those born vaginally. Infancy and childhood represent an opportunity to beneficially manipulate the microbiome through dietary or prebiotic interventions, which has the potential to affect both short- and long-term health outcomes.


Assuntos
Dieta , Fórmulas Infantis , Microbiota/fisiologia , Leite Humano/fisiologia , Prebióticos/administração & dosagem , Probióticos/administração & dosagem , Adolescente , Aleitamento Materno/tendências , Criança , Pré-Escolar , Dieta/tendências , Humanos , Lactente , Recém-Nascido , Leite Humano/microbiologia , Vigilância da População , Prebióticos/microbiologia
12.
PLoS One ; 15(2): e0229283, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32084202

RESUMO

Necrotizing enterocolitis (NEC) is a devastating intestinal emergency that affects ten percent of very low birth weight premature babies and costs society in both expense and heartache. It is probably caused by an inappropriate interaction of colonizing bacteria with an immature intestine. A possible preventative measure is to feed prematures their mother's expressed breast milk in conjunction with a probiotic. This synbiotic prevention reduces the severity and incidence of this condition. This study was designed to determine the mechanism of the synbiotic effect in human and mouse fetal intestine. Breast milk interacting with a NEC preventative probiotic such as Bifidobacterium infantis can produce increased levels of short chain fatty acids (acetate, propionate and butyrate) (SCFAs). SCFAs are known to be anti-inflammatory in mature enterocytes and immunocytes. Very little is known about their role in immature intestine. When exposed to a human fetal cell line, fetal intestinal organoids and fetal mouse intestine, these SCFAs were anti-inflammatory. Their mechanism of anti-inflammation differed from those reported for mature cells by involving the G-protein coupled receptor (GPR 109A) and inhibiting histone deacetylase 4 and 5. These bacterial metabolites may help explain the synbiotic anti-inflammatory effect of breast milk and probiotics given to premature infants at risk for NEC.


Assuntos
Bifidobacterium longum subspecies infantis/fisiologia , Enterócitos/citologia , Enterócitos/efeitos dos fármacos , Ácidos Graxos Voláteis/biossíntese , Ácidos Graxos Voláteis/farmacologia , Intestinos/microbiologia , Leite Humano/microbiologia , Animais , Anti-Inflamatórios/metabolismo , Anti-Inflamatórios/farmacologia , Enterócitos/metabolismo , Indução Enzimática/efeitos dos fármacos , Feto/microbiologia , Histona Desacetilases/genética , Humanos , Interleucina-1beta/metabolismo , Interleucina-8/metabolismo , Intestinos/citologia , Camundongos , Mutagênese Insercional/efeitos dos fármacos , Organoides/efeitos dos fármacos , Organoides/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo
13.
Benef Microbes ; 11(2): 151-162, 2020 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-31990220

RESUMO

Intestinal and human milk microbiota studies during infancy have shown variations according to geographical location, delivery mode, gestational age, and mother-related factors during pregnancy. In this study, we performed metagenomic mycobiota analyses of 44 transient and mature human milk among five different groups: mothers of normal spontaneous delivery-term (NS-T), caesarean delivery-term (CS-T), premature (PT), small for gestational age (SGA), and large for gestational age (LGA) infants. Fungi were detected in 80 out of the 88 samples. Regarding the number of observed fungal species, the NS-T group was more homogeneous (less variable) comparing the other groups (P<0.05). In the transient human milk samples, the most abundant species were Saccharomyces cerevisiae (33.3%) and Aspergillus glaucus (27.4%). While A. glaucus (33.7%) was second most abundant species in mature milk, S. cerevisiae disappeared (P<0.01) and Penicillium rubens became the most abundant species (35.5%) (P<0.05). Among the NS-T group, the most abundant species was Malassezia globosa in both transient and mature milk. In contrast, S. cerevisiae was the most abundant species in transient human milk (45.0%) in the CS-T group, but it disappeared in mature milk (P<0.01). In transient milk, M. globosa was only represented 6.0-9.0% of taxa in the PT, SGA, and LGA groups (P<0.05). In transient and mature milk in the PT, SGA and LGA groups, the most abundant species were A. glaucus and P. rubens. In mature milk samples, P. rubens is more abundant in CS-T group, PT group and LGA group, than the NS-T groups (P<0.05 for all). Although fungi constitute only a very small part of the human milk microbiome, we observed some changes that the human milk mycobiota composition varies in caesarean delivery, premature, SGA and LGA groups, comparing the normal spontaneous delivery, as well as differences between transient and mature human milk.


Assuntos
Peso ao Nascer , Parto Obstétrico/métodos , Idade Gestacional , Leite Humano/microbiologia , Micobioma , Adulto , Feminino , Fungos/isolamento & purificação , Humanos , Masculino , Mães , Ganho de Peso , Adulto Jovem
14.
PLoS One ; 15(1): e0219633, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31990909

RESUMO

Increasing evidence supports the importance of the breast milk microbiome in seeding the infant gut. However, the origin of bacteria in milk and the process of milk microbe-mediated seeding of infant intestine need further elucidation. Presumed sources of bacteria in milk include locations of mother-infant and mother-environment interactions. We investigate the role of mother-infant interaction on breast milk microbes. Shotgun metagenomics and 16S rRNA gene sequencing identified milk microbes of mother-infant pairs in breastfed infants and in infants that have never latched. Although breast milk has low overall biomass, milk microbes play an important role in seeding the infant gut. Breast milk bacteria were largely comprised of Staphylococcus, Streptococcus, Acinetobacter, and Enterobacter primarily derived from maternal areolar skin and infant oral sites in breastfeeding pairs. This suggests that the process of breastfeeding is a potentially important mechanism for propagation of breast milk microbes through retrograde flux via infant oral and areolar skin contact. In one infant delivered via Caesarian section, a distinct strain of Bifidobacteria breve was identified in maternal rectum, breast milk and the infant's stool potentially suggesting direct transmission. This may support the existence of microbial translocation of this anaerobic bacteria via the enteromammary pathway in humans, where maternal bacteria translocate across the maternal gut and are transferred to the mammary glands. Modulating sources of human milk microbiome seeding potentially imply opportunities to ultimately influence the development of the infant microbiome and health.


Assuntos
Translocação Bacteriana , Bifidobacterium breve/isolamento & purificação , Intestinos/microbiologia , Glândulas Mamárias Humanas/microbiologia , Leite Humano/microbiologia , Acinetobacter/genética , Acinetobacter/isolamento & purificação , Adolescente , Adulto , Bifidobacterium breve/genética , Enterobacter/genética , Enterobacter/isolamento & purificação , Feminino , Microbioma Gastrointestinal/genética , Humanos , Lactente , Recém-Nascido , Metagenômica/métodos , Gravidez , RNA Ribossômico 16S/genética , Staphylococcus/genética , Staphylococcus/isolamento & purificação , Streptococcus/genética , Streptococcus/isolamento & purificação
15.
Sci Rep ; 10(1): 123, 2020 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-31924794

RESUMO

In addition to providing nutritional and bioactive factors necessary for infant development, human breast milk contains bacteria that contribute to the establishment of commensal microbiota in the infant. However, the composition of this bacterial community differs considerably between studies. We hypothesised that bacterial DNA extraction methodology from breast milk samples are a substantial contributor to these inter-study differences. We tested this hypothesis by applying five widely employed methodologies to a mock breast milk sample and four individual human breast milk samples. Significant differences in DNA yield and purity were observed between methods (P < 0.05). Microbiota composition, assessed by 16S rRNA gene amplicon sequencing, also differed significantly with extraction methodology (P < 0.05), including in the contribution of contaminant signal. Concerningly, many of the bacterial taxa identified here as contaminants have been reported as components of the breast milk microbiome in other studies. These findings highlight the importance of using stringent, well-validated, DNA extraction methodologies for analysis of the breast milk microbiome, and exercising caution interpreting microbiota data from low-biomass contexts.


Assuntos
Fracionamento Químico/métodos , DNA Bacteriano/isolamento & purificação , Microbiota , Leite Humano/microbiologia , DNA Bacteriano/genética , Humanos , Análise de Sequência de DNA
16.
Nutrients ; 12(1)2020 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-31936487

RESUMO

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.


Assuntos
Aderência Bacteriana/fisiologia , Leite Humano , Probióticos , Bifidobacterium/fisiologia , Fezes/química , Fezes/microbiologia , Feminino , Humanos , Lactente , Recém-Nascido , Lactobacillus rhamnosus/fisiologia , Bancos de Leite , Leite Humano/química , Leite Humano/microbiologia , Leite Humano/fisiologia , Muco/química
17.
Food Funct ; 11(1): 435-447, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31829373

RESUMO

The emergence of multi-drug resistant (MDR) pathogens greatly challenges the development of new drugs. Probiotics with the ability to inhibit MDR pathogens offer advantages over chemical antibiotics and drugs due to increased safety and fewer side effects. This study reports that Lactobacillus rhamnosus SHA113 (isolated from breast milk) significantly inhibited MDR Escherichia coli both in vitro and in vivo. MDR E. coli caused more severe inflammatory effects. TNF-α and IL-6 levels increased, while the IL-10 content decreased in serum. MDR E. coli caused disturbance in the gut microbial balance, increased the total coliform, decreased lactic acid bacteria in feces, decreased Firmicutes, and increased both Bacteroidetes and Proteobacteria. At the end of the curing treatment, ampicillin (AMP) treatment significantly reduced lactic acid bacteria compared to total coliform in feces and exacerbated the increase of Proteobacteria caused by MDR E. coli. L. rhamnosus SHA113 treatment resulted in a more significant and faster decrease of total coliform in feces and a significant decrease of Proteobacteria in the gut microbiota. The increase of total coliform in feces (caused by MDR E. coli infection) was positively correlated with IL-6 and TNF-α and negatively correlated with IL-10 in serum. However, the increase of lactic acid bacteria in feces (caused by L. rhamnosus SHA113 treatment) was negatively correlated with serum TNF-α, indicating that SHA113 exerted anti-inflammatory effects. These results suggest that L. rhamnosus SHA113 has great potential for inhibiting infections by MDR E. coli and for regulating the gut flora balance.


Assuntos
Infecções por Escherichia coli/terapia , Microbioma Gastrointestinal , Lactobacillus rhamnosus/isolamento & purificação , Leite Humano/microbiologia , Probióticos , Animais , Farmacorresistência Bacteriana Múltipla , Escherichia coli , Fezes/microbiologia , Humanos , Interleucina-10/sangue , Interleucina-6/sangue , Masculino , Camundongos , Fator de Necrose Tumoral alfa/sangue
18.
J Perinat Med ; 48(2): 179-183, 2020 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-31874102

RESUMO

Background To meet the nutritional needs of preterm infants, multicomponent nutrient fortifiers are added to human milk. The fortified human milk (FHM) product changes the physical and biochemical characteristics of the milk. We questioned whether such physical-chemical changes in the milk would alter intrinsic probiotic bacterial activity. The objective of the study was to evaluate the effect of osmolality and pH on the growth of probiotic bacterial species intrinsic to human milk. Methods Human milk samples (n = 26) were collected from mothers in the neonatal intensive care unit (NICU) and stored at -20°C until analyzed. The samples were thawed and divided into three portions. Human milk fortifiers (HMFs) were added to two portions to prepare concentrations of FHM. The remaining portion was the unfortified control sample. Each sample was then divided into two parts. One part (baseline) was used to measure the osmolality and pH and plated on selective agar to enumerate the growth of lactobacilli and bifidobacteria species. The remaining part was incubated at 37°C for 24 h to further test bacterial integrity (post-incubation) and then the same measurements were made (osmolality, pH, bacterial colony counts). Results When compared with unfortified milk at baseline, osmolality increased and pH decreased significantly after the addition of HMFs. Lactobacilli and bifidobacteria colony counts did not differ among the groups pre-incubation. Post-incubation lactobacilli and bifidobacteria increased in all the groups. Conclusion The appropriate addition of HMFs differentially affected the osmolality and pH of the milk. These physical changes did not affect the growth of probiotic bacterial species.


Assuntos
Alimentos Fortificados/microbiologia , Leite Humano/microbiologia , Probióticos , Bifidobacterium/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio , Lactobacillus/crescimento & desenvolvimento , Leite Humano/química , Concentração Osmolar
19.
Sci Rep ; 9(1): 18653, 2019 12 09.
Artigo em Inglês | MEDLINE | ID: mdl-31819085

RESUMO

Archaeal sequences have been detected in human colostrum and milk, but no studies have determined whether living archaea are present in either of these fluids. Methanogenic archaea are neglected since they are not detected by usual molecular and culture methods. By using improved DNA detection protocols and microbial culture techniques associated with antioxidants previously developed in our center, we investigated the presence of methanogenic archaea using culture and specific Methanobrevibacter smithii and Methanobrevibacter oralis real-time PCR in human colostrum and milk. M. smithii was isolated from 3 colostrum and 5 milk (day 10) samples. M. oralis was isolated from 1 milk sample. For 2 strains, the genome was sequenced, and the rhizome was similar to that of strains previously isolated from the human mouth and gut. M. smithii was detected in the colostrum or milk of 5/13 (38%) and 37/127 (29%) mothers by culture and qPCR, respectively. The different distribution of maternal body mass index according to the detection of M. smithii suggested an association with maternal metabolic phenotype. M. oralis was not detected by molecular methods. Our results suggest that breastfeeding may contribute to the vertical transmission of these microorganisms and may be essential to seed the infant's microbiota with these neglected critical commensals from the first hour of life.


Assuntos
Aleitamento Materno/efeitos adversos , Colostro/microbiologia , Methanobrevibacter/isolamento & purificação , Leite Humano/microbiologia , Animais , Índice de Massa Corporal , Crescimento Quimioautotrófico/genética , DNA Arqueal/genética , DNA Arqueal/isolamento & purificação , Euryarchaeota/genética , Euryarchaeota/patogenicidade , Fezes/microbiologia , Feminino , Humanos , Lactente , Methanobrevibacter/genética , Methanobrevibacter/patogenicidade , Microbiota/genética , Mães , Gravidez
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