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1.
Cell ; 185(23): 4280-4297.e12, 2022 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-36323316

RESUMO

The gut microbiome has an important role in infant health and development. We characterized the fecal microbiome and metabolome of 222 young children in Dhaka, Bangladesh during the first two years of life. A distinct Bifidobacterium longum clade expanded with introduction of solid foods and harbored enzymes for utilizing both breast milk and solid food substrates. The clade was highly prevalent in Bangladesh, present globally (at lower prevalence), and correlated with many other gut taxa and metabolites, indicating an important role in gut ecology. We also found that the B. longum clades and associated metabolites were implicated in childhood diarrhea and early growth, including positive associations between growth measures and B. longum subsp. infantis, indolelactate and N-acetylglutamate. Our data demonstrate geographic, cultural, seasonal, and ecological heterogeneity that should be accounted for when identifying microbiome factors implicated in and potentially benefiting infant development.


Assuntos
Bifidobacterium longum , Lactente , Criança , Feminino , Humanos , Pré-Escolar , Bifidobacterium longum/metabolismo , Bifidobacterium/metabolismo , Desmame , Oligossacarídeos/metabolismo , Bangladesh , Leite Humano , Fezes/microbiologia
2.
Appl Environ Microbiol ; 85(24)2019 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-31585993

RESUMO

Single-nucleotide polymorphisms (SNPs) are widely used for whole-genome sequencing (WGS)-based subtyping of foodborne pathogens in outbreak and source tracking investigations. Mobile genetic elements (MGEs) are commonly present in bacterial genomes and may affect SNP subtyping results if their evolutionary history and dynamics differ from that of the bacterial chromosomes. Using Salmonella enterica as a model organism, we surveyed major categories of MGEs, including plasmids, phages, insertion sequences, integrons, and integrative and conjugative elements (ICEs), in 990 genomes representing 21 major serotypes of S. enterica We evaluated whether plasmids and chromosomal MGEs affect SNP subtyping with 9 outbreak clusters of different serotypes found in the United States in 2018. The median total length of chromosomal MGEs accounted for 2.5% of a typical S. enterica chromosome. Of the 990 analyzed S. enterica isolates, 68.9% contained at least one assembled plasmid sequence. The median total length of assembled plasmids in these isolates was 93,671 bp. Plasmids that carry high densities of SNPs were found to substantially affect both SNP phylogenies and SNP distances among closely related isolates if they were present in the reference genome for SNP subtyping. In comparison, chromosomal MGEs were found to have limited impact on SNP subtyping. We recommend the identification of plasmid sequences in the reference genome and the exclusion of plasmid-borne SNPs from SNP subtyping analysis.IMPORTANCE Despite increasingly routine use of WGS and SNP subtyping in outbreak and source tracking investigations, whether and how MGEs affect SNP subtyping has not been thoroughly investigated. Besides chromosomal MGEs, plasmids are frequently entangled in draft genome assemblies and yet to be assessed for their impact on SNP subtyping. This study provides evidence-based guidance on the treatment of MGEs in SNP analysis for Salmonella to infer phylogenetic relationship and SNP distance between isolates.


Assuntos
Sequências Repetitivas Dispersas/genética , Polimorfismo de Nucleotídeo Único , Salmonella enterica/classificação , Salmonella enterica/genética , Cromossomos Bacterianos , Surtos de Doenças , Genoma Bacteriano , Humanos , Filogenia , Plasmídeos/isolamento & purificação , Sorogrupo , Sequenciamento Completo do Genoma
4.
J Pediatr Gastroenterol Nutr ; 63(6): 681-687, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27035371

RESUMO

OBJECTIVES: Microbiota modulation by probiotics in infants born by cesarean (C)-section is poorly understood. We aimed at assessing the response of C-section-delivered infant microbiota to a formula containing Lactobacillus reuteri Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSM) 17938 and comparing it with that of vaginally delivered infants. METHODS: Infants delivered by C-section (C) and vaginally (V) were randomized to receive either control formula (CCt, n = 10; VCt, n = 10) or the same formula containing L reuteri (CLr, n = 11; VLr, n = 9) within 72 hours following birth. Stool samples were collected at 2 weeks and 4 months of age. Microbial DNA was extracted, amplified, and pyrosequenced. RESULTS: The phylogenetic profiles of the CLr, VCt, and VLr microbiota were not significantly different at any age but diverged from that of CCt at 2 weeks. Compared with VCt, CCt displayed lower Bifidobacterium and higher Enterobacter, unclassified Enterobacteriaceae, Enterococcus, Clostridium, and unclassified Clostridiaceae relative abundance at 2 weeks, as well as lower Collinsella and higher Enterococcus and Coprococcus abundance at 4 months. The level of most of these taxa was not significantly different between the CLr and the vaginal-delivery groups. Compared with VCt, the only difference observed in VLr microbiota was higher Lactobacillus at the 2 study ages and Coprococcus at 4 months. CONCLUSIONS: Our results show that a formula containing L reuteri DSM 17938 does not essentially alter the microbiota in vaginally born infants. In C-section-delivered infants, however, this strain seems to play the role of keystone species by modulating the early development of the microbiota toward the composition found after vaginal delivery.


Assuntos
Cesárea/efeitos adversos , Fezes/microbiologia , Fórmulas Infantis , Microbiota , Probióticos/administração & dosagem , Contagem de Colônia Microbiana , Feminino , Idade Gestacional , Humanos , Lactente , Recém-Nascido , Limosilactobacillus reuteri/crescimento & desenvolvimento , Gravidez , Estudos Prospectivos
5.
J Clin Microbiol ; 52(5): 1590-4, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24599973

RESUMO

Our study is the first to compare the nasopharyngeal microbiota of pediatric pneumonia patients and control children by 454 pyrosequencing. A distinct microbiota was associated with different pneumonia etiologies. Viral pneumonia was associated with a high abundance of the operational taxonomic unit (OTU) corresponding to Moraxella lacunata. Patients with nonviral pneumonia showed high abundances of OTUs of three typical bacterial pathogens, Streptococcus pneumoniae complex, Haemophilus influenzae complex, and Moraxella catarrhalis. Patients classified as having no definitive etiology harbored microbiota particularly enriched in the H. influenzae complex. We did not observe a commensal taxon specifically associated with health. The microbiota of the healthy nasopharynx was more diverse and contained a wider range of less abundant taxa.


Assuntos
Microbiota/genética , Nasofaringe/microbiologia , Nasofaringe/virologia , Pneumonia/microbiologia , Pneumonia/virologia , Adolescente , Estudos de Casos e Controles , Criança , Pré-Escolar , Infecções por Haemophilus/diagnóstico , Infecções por Haemophilus/microbiologia , Haemophilus influenzae/genética , Humanos , Lactente , Moraxella catarrhalis/genética , Infecções por Moraxellaceae/diagnóstico , Infecções por Moraxellaceae/microbiologia , Infecções Pneumocócicas/diagnóstico , Infecções Pneumocócicas/microbiologia , Pneumonia/diagnóstico , Estudos Prospectivos , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/microbiologia , Infecções Respiratórias/virologia , Streptococcus pneumoniae/genética
7.
Brief Bioinform ; 13(6): 747-50, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22543244

RESUMO

The human gut is populated by a rich and diverse microbiota that has been described as a human metabolic organ. Its composition has an impact on the health status of the host and could be the target or effector of dietary health effects. The advent of high-throughput sequencing technologies and their integration with advanced analysis methods enables the development of new approaches to characterize the gut microbiota composition and initiates the understanding of its functionality. These technological aspects are not necessarily the limiting factor to successfully identify biological correlations between gut microbiota, diet and health status, as there are other very important aspects. The aim of this article is to address the gut microbiota characterization methodologies, highlight some advantages and challenges, and give our opinion on how critical the sampling and the study design are.


Assuntos
Trato Gastrointestinal/microbiologia , Metagenoma/genética , Dieta , Humanos , Metagenômica
8.
Br J Nutr ; 111(8): 1507-19, 2014 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-24299712

RESUMO

The present study investigated the impact of a Lactobacillus rhamnosus CGMCC1.3724 (LPR) supplementation on weight loss and maintenance in obese men and women over 24 weeks. In a double-blind, placebo-controlled, randomised trial, each subject consumed two capsules per d of either a placebo or a LPR formulation (1.6 × 10(8) colony-forming units of LPR/capsule with oligofructose and inulin). Each group was submitted to moderate energy restriction for the first 12 weeks followed by 12 weeks of weight maintenance. Body weight and composition were measured at baseline, at week 12 and at week 24. The intention-to-treat analysis showed that after the first 12 weeks and after 24 weeks, mean weight loss was not significantly different between the LPR and placebo groups when all the subjects were considered. However, a significant treatment × sex interaction was observed. The mean weight loss in women in the LPR group was significantly higher than that in women in the placebo group (P = 0.02) after the first 12 weeks, whereas it was similar in men in the two groups (P= 0.53). Women in the LPR group continued to lose body weight and fat mass during the weight-maintenance period, whereas opposite changes were observed in the placebo group. Changes in body weight and fat mass during the weight-maintenance period were similar in men in both the groups. LPR-induced weight loss in women was associated not only with significant reductions in fat mass and circulating leptin concentrations but also with the relative abundance of bacteria of the Lachnospiraceae family in faeces. The present study shows that the Lactobacillus rhamnosus CGMCC1.3724 formulation helps obese women to achieve sustainable weight loss.


Assuntos
Lacticaseibacillus rhamnosus , Obesidade/tratamento farmacológico , Probióticos/uso terapêutico , Redução de Peso , Tecido Adiposo/metabolismo , Adulto , Colo/microbiologia , Suplementos Nutricionais , Método Duplo-Cego , Ingestão de Energia , Fezes , Feminino , Humanos , Análise de Intenção de Tratamento , Leptina/sangue , Masculino , Pessoa de Meia-Idade , Obesidade/sangue , Obesidade/metabolismo , Fatores Sexuais , Adulto Jovem
10.
Microorganisms ; 10(2)2022 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-35208658

RESUMO

Strains of Bifidobacterium longum subsp. infantis (B. infantis) are amongst the first to colonize the infant gut, partly due to their capacity to metabolize complex human milk oligosaccharides (HMO), and are proposed to play a key role in the development of the infant gut. Since early life, B. infantis supplementation is of high interest, and detailed phylogenetic, functional and safety characterization of the selected strains should be pursued. Using a combination of long and short-read sequencing technologies, we first decipher the genetic distance between different isolates of the same B. infantis strain. Using the same approach, we show that several publicly available genomes recapitulate this strain-level distance as compared to two of the first strains obtained in the 1950s. Furthermore, we demonstrate that the two 1950s B. infantis strains display different functional and safety attributes, as ATCC 15697 is resistant to streptomycin and shows a preference towards lacto-N-tetraose LNT and sialylated HMOs, while LMG 11588 is sensitive to all tested antibiotics and shows a preference towards fucosylated HMOs. Overall, our work highlights that the current diversity observed in B. infantis is likely underestimated and that strain selection within this subspecies must be the subject of scientific pursuit and associated evaluation.

11.
Genes (Basel) ; 12(2)2021 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-33671973

RESUMO

Whole genome sequencing (WGS) has arisen as a powerful tool to perform pathogen source tracking in the food industry thanks to several developments in recent years. However, the cost associated to this technology and the degree of expertise required to accurately process and understand the data has limited its adoption at a wider scale. Additionally, the time needed to obtain actionable information is often seen as an impairment for the application and use of the information generated via WGS. Ongoing work towards standardization of wet lab including sequencing protocols, following guidelines from the regulatory authorities and international standardization efforts make the technology more and more accessible. However, data analysis and results interpretation guidelines are still subject to initiatives coming from distinct groups and institutions. There are multiple bioinformatics software and pipelines developed to handle such information. Nevertheless, little consensus exists on a standard way to process the data and interpret the results. Here, we want to present the constraints we face in an industrial setting and the steps we consider necessary to obtain high quality data, reproducible results and a robust interpretation of the obtained information. All of this, in a time frame allowing for data-driven actions supporting factories and their needs.


Assuntos
Indústria Alimentícia/tendências , Microbiologia de Alimentos/tendências , Genoma Bacteriano/genética , Sequenciamento Completo do Genoma , Biologia Computacional , Análise de Dados , Humanos , Software
12.
Nucleic Acids Res ; 36(Database issue): D485-90, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17981845

RESUMO

New methods and tools are needed to exploit the unprecedented source of information made available by the completed and ongoing whole genome sequencing projects. The Narcisse database is dedicated to the study of genome conservation, from sequence similarities to conserved chromosomal segments or conserved syntenies, for a large number of animals, plants and bacterial completely sequenced genomes. The query interface, a comparative genome browser, enables to navigate between genome dotplots, comparative maps and sequence alignments. The Narcisse database can be accessed at http://narcisse.toulouse.inra.fr.


Assuntos
Bases de Dados de Ácidos Nucleicos , Sintenia , Animais , Sequência de Bases , Mapeamento Cromossômico , Sequência Conservada , Genoma Bacteriano , Genoma de Planta , Genômica , Humanos , Internet , Ratos , Alinhamento de Sequência , Interface Usuário-Computador
13.
Front Microbiol ; 9: 446, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29593690

RESUMO

Whole genome sequencing (WGS), using high throughput sequencing technology, reveals the complete sequence of the bacterial genome in a few days. WGS is increasingly being used for source tracking, pathogen surveillance and outbreak investigation due to its high discriminatory power. In the food industry, WGS used for source tracking is beneficial to support contamination investigations. Despite its increased use, no standards or guidelines are available today for the use of WGS in outbreak and/or trace-back investigations. Here we present a validation of our complete (end-to-end) WGS workflow for Listeria monocytogenes and Salmonella enterica including: subculture of isolates, DNA extraction, sequencing and bioinformatics analysis. This end-to-end WGS workflow was evaluated according to the following performance criteria: stability, repeatability, reproducibility, discriminatory power, and epidemiological concordance. The current study showed that few single nucleotide polymorphism (SNPs) were observed for L. monocytogenes and S. enterica when comparing genome sequences from five independent colonies from the first subculture and five independent colonies after the tenth subculture. Consequently, the stability of the WGS workflow for L. monocytogenes and S. enterica was demonstrated despite the few genomic variations that can occur during subculturing steps. Repeatability and reproducibility were also demonstrated. The WGS workflow was shown to have a high discriminatory power and has the ability to show genetic relatedness. Additionally, the WGS workflow was able to reproduce published outbreak investigation results, illustrating its capability of showing epidemiological concordance. The current study proposes a validation approach comprising all steps of a WGS workflow and demonstrates that the workflow can be applied to L. monocytogenes or S. enterica.

14.
J Bacteriol ; 189(22): 8206-14, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17693496

RESUMO

Numerous T4-like Escherichia coli phages were isolated from human stool and environmental wastewater samples in Bangladesh and Switzerland. The sequences of the major head gene (g23) revealed that these coliphages could be placed into four subgroups, represented by the phages T4, RB69, RB49, and JS98. Thus, JS98 defines a new major subgroup of E. coli T4-like phages. We conducted an analysis of the 169-kb JS98 genome sequence. Overall, 198 of the 266 JS98 open reading frames (ORFs) shared amino acid sequence identity with the reference T4 phage, 41 shared identity with other T4-like phages, and 27 ORFs lacked any database matches. Genes on the plus strand encoded virion proteins, which showed moderate to high sequence identity with T4 proteins. The right genome half of JS98 showed a higher degree of sequence conservation with T4 and RB69, even for the nonstructural genes, than did the left genome half, containing exclusively nonstructural genes. Most of the JS98-specific genes were found in the left genome half. Two came as a hypervariability cluster, but most represented isolated genes, suggesting that they were acquired separately in multiple acquisition events. No evidence for DNA exchange between JS98 phage and the E. coli host genome or coliphages other than T4 was observed. No undesired genes which could compromise its medical use were detected in the JS98 genome sequence.


Assuntos
Bacteriófago T4/classificação , Bacteriófago T4/genética , Escherichia coli/virologia , Genoma Viral , DNA Viral/genética , Regulação Viral da Expressão Gênica , Dados de Sequência Molecular , Filogenia , Proteínas Virais/genética , Proteínas Virais/metabolismo
15.
Genome Announc ; 4(2)2016 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-27013041

RESUMO

Cronobacteris associated with infant infections and the consumption of reconstituted infant formula. Here we sequenced and closed six genomes ofC. condimenti(T),C. muytjensii(T),C. universalis(T),C. malonaticus(T),C. dublinensis(T), andC. sakazakiithat can be used as reference genomes in single nucleotide polymorphism (SNP)-based next-generation sequencing (NGS) analysis for source tracking investigations.

16.
Genome Announc ; 4(6)2016 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-27856580

RESUMO

Lactobacillus fermentum NCC2970 (CNCM I-5068) is a lactic acid bacterium originating from the Nestle Culture Collection. Here, we disclose its full 1.9-Gb genome sequence comprising one chromosome with no plasmid.

17.
EBioMedicine ; 4: 124-37, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26981577

RESUMO

BACKGROUND: Antibiotic resistance is rising in important bacterial pathogens. Phage therapy (PT), the use of bacterial viruses infecting the pathogen in a species-specific way, is a potential alternative. METHOD: T4-like coliphages or a commercial Russian coliphage product or placebo was orally given over 4 days to Bangladeshi children hospitalized with acute bacterial diarrhea. Safety of oral phage was assessed clinically and by functional tests; coliphage and Escherichia coli titers and enteropathogens were determined in stool and quantitative diarrhea parameters (stool output, stool frequency) were measured. Stool microbiota was studied by 16S rRNA gene sequencing; the genomes of four fecal Streptococcus isolates were sequenced. FINDINGS: No adverse events attributable to oral phage application were observed (primary safety outcome). Fecal coliphage was increased in treated over control children, but the titers did not show substantial intestinal phage replication (secondary microbiology outcome). 60% of the children suffered from a microbiologically proven E. coli diarrhea; the most frequent diagnosis was ETEC infections. Bacterial co-pathogens were also detected. Half of the patients contained phage-susceptible E. coli colonies in the stool. E. coli represented less than 5% of fecal bacteria. Stool ETEC titers showed only a short-lived peak and were otherwise close to the replication threshold determined for T4 phage in vitro. An interim analysis after the enrollment of 120 patients showed no amelioration in quantitative diarrhea parameter by PT over standard care (tertiary clinical outcome). Stool microbiota was characterized by an overgrowth with Streptococcus belonging to the Streptococcus gallolyticus and Streptococcus salivarius species groups, their abundance correlated with quantitative diarrhea outcome, but genome sequencing did not identify virulence genes. INTERPRETATION: Oral coliphages showed a safe gut transit in children, but failed to achieve intestinal amplification and to improve diarrhea outcome, possibly due to insufficient phage coverage and too low E. coli pathogen titers requiring higher oral phage doses. More knowledge is needed on in vivo phage-bacterium interaction and the role of E. coli in childhood diarrhea for successful PT. FUNDING: The study was supported by a grant from Nestlé Nutrition and Nestlé Health Science. The trial was registered with Identifier NCT00937274 at ClinicalTrials.gov.


Assuntos
Colífagos/patogenicidade , Diarreia/terapia , Infecções por Escherichia coli/terapia , Terapia por Fagos , Administração Oral , Adolescente , Bangladesh , Criança , Diarreia/microbiologia , Escherichia coli Enteropatogênica/virologia , Infecções por Escherichia coli/microbiologia , Feminino , Humanos , Masculino
18.
Gut Microbes ; 6(5): 321-5, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26516657

RESUMO

The gut of the human neonate is colonized rapidly after birth from an early sparse and highly distinct microbiota to a more adult-like and convergent state, within 1 to 3 years. The progression of colonizing bacterial species is non-random. During the first months of life several shifts commonly occur in the species prevalent in our guts. Although the sequential progression of these species is remarkably consistent across individuals and geographies, there is inter-individual variation in the rate of progression. Our study and others suggest that the rate is influenced by environmental factors, and influences our future health. In this article, we review our recent contribution to cataloging the developing infant gut microbiota alongside other important recent studies. We suggest testable hypotheses that arise from this synthesis.


Assuntos
Microbioma Gastrointestinal , Saúde , Intestinos/microbiologia , Humanos , Lactente , Recém-Nascido , Fatores de Tempo
19.
mBio ; 6(1)2015 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-25650398

RESUMO

UNLABELLED: We found that the relatively simple microbiota of young infants shifts predictably to a more mature anaerobic microbiota during infancy and the dynamics of this shift are influenced by environmental factors. In this longitudinal study of 75 infants, we demonstrate high interindividual variability within the normal range of birth outcomes, especially in the rate of microbiota progression. Most had acquired a microbiota profile high in Bifidobacterium and Collinsella by 6 months of age, but the time point of this acquisition was later in infants delivered by caesarean section and those born after a shorter duration of gestation. Independently of the delivery mode and gestation duration, infants who acquired a profile high in Bifidobacterium and Collinsella at a later age had lower adiposity at 18 months of age. IMPORTANCE: This study shows that the acquisition of the early microbiota is strongly influenced by environmental factors such as the delivery mode and duration of gestation, even in healthy neonates. The composition of the early microbiota has been linked with long-lasting effects on health and disease. Here we show that the rate of acquisition of certain microbiota predicts adiposity at 18 months of age and so potentially the risk of later obesity.


Assuntos
Adiposidade , Bactérias/isolamento & purificação , Microbioma Gastrointestinal , Intestinos/microbiologia , Obesidade/microbiologia , Bactérias/classificação , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Biodiversidade , Feminino , Humanos , Lactente , Recém-Nascido , Estudos Longitudinais , Masculino , Obesidade/fisiopatologia
20.
Nat Med ; 20(2): 159-66, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24390308

RESUMO

Metabolites from intestinal microbiota are key determinants of host-microbe mutualism and, consequently, the health or disease of the intestinal tract. However, whether such host-microbe crosstalk influences inflammation in peripheral tissues, such as the lung, is poorly understood. We found that dietary fermentable fiber content changed the composition of the gut and lung microbiota, in particular by altering the ratio of Firmicutes to Bacteroidetes. The gut microbiota metabolized the fiber, consequently increasing the concentration of circulating short-chain fatty acids (SCFAs). Mice fed a high-fiber diet had increased circulating levels of SCFAs and were protected against allergic inflammation in the lung, whereas a low-fiber diet decreased levels of SCFAs and increased allergic airway disease. Treatment of mice with the SCFA propionate led to alterations in bone marrow hematopoiesis that were characterized by enhanced generation of macrophage and dendritic cell (DC) precursors and subsequent seeding of the lungs by DCs with high phagocytic capacity but an impaired ability to promote T helper type 2 (TH2) cell effector function. The effects of propionate on allergic inflammation were dependent on G protein-coupled receptor 41 (GPR41, also called free fatty acid receptor 3 or FFAR3), but not GPR43 (also called free fatty acid receptor 2 or FFAR2). Our results show that dietary fermentable fiber and SCFAs can shape the immunological environment in the lung and influence the severity of allergic inflammation.


Assuntos
Bacteroidetes/metabolismo , Fibras na Dieta/microbiologia , Hematopoese/fisiologia , Hipersensibilidade/fisiopatologia , Intestinos/microbiologia , Microbiota/fisiologia , Transferência Adotiva , Animais , Sequência de Bases , Citocinas/metabolismo , Primers do DNA/genética , DNA Bacteriano/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Ácidos Graxos/sangue , Fezes/química , Feminino , Citometria de Fluxo , Pulmão/química , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Estatísticas não Paramétricas
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