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1.
Mar Genomics ; 66: 100991, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36116403

RESUMO

The aim of the current work was to investigate the impact of marine aquaculture on seafloor biogeochemistry and diversity from pristine environments in the northern part of Norway. Our analytical approach included analyses of 182 samples from 16 aquaculture sites using 16S and 18S rRNA, shotgun analyses, visual examination of macro-organisms, in addition to chemical measurements. We observed a clear bimodal distribution of the prokaryote composition and richness, determined by analyses of 16S rRNA gene operational taxonomic units (OTUs). The high OTU richness cluster was associated with non-perturbed environments and farness from the aquaculture sites, while the low OTU richness cluster was associated with perturbed environments and proximity to the aquaculture sites. Similar patterns were also observed for eukaryotes using 18S rRNA gene analyses and visual examination, but without a bimodal distribution of OTU richness. Shotgun sequencing showed the archaeum Nitrosopumilus as dominant for the high OTU richness cluster, and the epsilon protobacterium Sulfurovum as dominant for the low OTU richness cluster. Metabolic reconstruction of Nitrosopumilus indicates nitrification as the main metabolic pathway. Sulfurovum, on the other hand, was associated with sulfur oxidation and denitrification. Changes in nitrogen and sulfur metabolism is proposed as a potential explanation for the difference between the high and low OTU richness clusters. In conclusion, these findings suggest that pollution from elevated loads of organic waste drives the microbiota towards a complete alteration of respiratory routes and species composition, in addition to a collapse in prokaryote OTU richness.


Assuntos
Microbiota , RNA Ribossômico 16S/genética , Aquicultura , Enxofre , Noruega
2.
Br J Dermatol ; 186(1): 50-58, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-33511639

RESUMO

BACKGROUND: Diagnosing atopic dermatitis (AD) in infants is challenging. OBJECTIVES: To determine the incidence and persistence of eczema and AD in infants using the UK Working Party (UKWP) and Hanifin and Rajka (H&R) criteria. METHODS: A cohort of 1834 infants was examined clinically at 3, 6 and 12 months of age. AD was diagnosed by UKWP (3, 6 and 12 months) and H&R (12 months) criteria. Logistic regression models were used to assess the relationship between AD and eczema. RESULTS: Eczema was observed in 628 (34·2%) infants (n = 240, n = 359 and n = 329 at 3, 6 and 12 months, respectively), with AD diagnosed in 212 (33·7%) infants with any eczema and in 64/78 (82%) infants with eczema at all three visits. The odds of AD were lower with first presentation of eczema at 6 [odds ratio (OR) 0·33, 95% confidence interval (CI) 0·22-0·48] or 12 months (OR 0·49, 95% CI 0·32-0·74) than at 3 months, and higher in infants with eczema at three (OR 23·1, 95% CI 12·3-43·6) or two (OR 6·5, 95% CI 4·3-9·9) visits vs. one visit only. At 12 months, 156/329 (47·4%) fulfilled the UKWP and/or H&R criteria; 27 (8%) fulfilled the UKWP criteria only and 65 (20%) only the H&R criteria. Of the 129 infants who fulfilled the H&R criteria, 44 (34·1%) did not meet the itch criterion. CONCLUSIONS: Used in combination and at multiple timepoints, the UKWP and H&R criteria for AD may be useful in clinical research but may have limited value in most other clinical settings.


Assuntos
Dermatite Atópica , Eczema , Estudos de Coortes , Dermatite Atópica/diagnóstico , Dermatite Atópica/epidemiologia , Eczema/diagnóstico , Eczema/epidemiologia , Humanos , Incidência , Lactente , Prurido
3.
Br J Dermatol ; 186(3): 544-552, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34698386

RESUMO

BACKGROUND: Loss-of-function mutations in the skin barrier gene filaggrin (FLG) increase the risk of atopic dermatitis (AD), but their role in skin barrier function, dry skin and eczema in infancy is unclear. OBJECTIVES: To determine the role of FLG mutations in impaired skin barrier function, dry skin, eczema and AD at 3 months of age and throughout infancy. METHODS: FLG mutations were analysed in 1836 infants in the Scandinavian population-based PreventADALL study. Transepidermal water loss (TEWL), dry skin, eczema and AD were assessed at 3, 6 and 12 months of age. RESULTS: FLG mutations were observed in 166 (9%) infants. At 3 months, carrying FLG mutations was not associated with impaired skin barrier function (TEWL > 11·3 g m-2  h-1 ) or dry skin, but was associated with eczema [odds ratio (OR) 2·89, 95% confidence interval (CI) 1·95-4·28; P < 0·001]. At 6 months, mutation carriers had significantly higher TEWL than nonmutation carriers [mean 9·68 (95% CI 8·69-10·68) vs. 8·24 (95% CI 7·97-8·15), P < 0·01], and at 3 and 6 months mutation carriers had an increased risk of dry skin on the trunk (OR 1·87, 95% CI 1·25-2·80; P = 0·002 and OR 2·44, 95% CI 1·51-3·95; P < 0·001) or extensor limb surfaces (OR 1·52, 95% CI 1·04-2·22; P = 0·028 and OR 1·74, 95% CI 1·17-2·57; P = 0·005). FLG mutations were associated with eczema and AD in infancy. CONCLUSIONS: FLG mutations were not associated with impaired skin barrier function or dry skin in general at 3 months of age, but increased the risk for eczema, and for dry skin on the trunk and extensor limb surfaces at 3 and 6 months.


Assuntos
Dermatite Atópica , Eczema , Proteínas Filagrinas/genética , Dermatite Atópica/genética , Eczema/genética , Genótipo , Humanos , Lactente , Proteínas de Filamentos Intermediários/genética , Proteínas de Filamentos Intermediários/metabolismo , Mutação/genética , Pele/metabolismo
4.
Microbiome ; 8(1): 160, 2020 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-33198805

RESUMO

The gut microbiome plays a key role in animal health and metabolism through the intricate functional interconnection between the feed, gut microbes, and the host. Unfortunately, in aquaculture, the links between gut microbes and fish genetics and production phenotypes are not well understood.In this study, we investigate the associations between gut microbial communities, fish feed conversion, and fish genetics in the domestic Atlantic salmon. Microbial community composition was determined for 230 juvenile fish from 23 full-sib families and was then regressed on growth, carbon and nitrogen metabolism, and feed efficiency. We only found weak associations between host genetics and microbial composition. However, we did identify significant (p < 0.05) associations between the abundance of three microbial operational taxonomical units (OTUs) and fish metabolism phenotypes. Two OTUs were associated with both carbon metabolism in adipose tissue and feed efficiency, while a third OTU was associated with weight gain.In conclusion, this study demonstrates an intriguing association between host lipid metabolism and the gut microbiota composition in Atlantic salmon. Video Abstract.


Assuntos
Envelhecimento/metabolismo , Microbioma Gastrointestinal , Salmo salar/metabolismo , Salmo salar/microbiologia , Ração Animal , Animais , Aquicultura , Feminino , Microbioma Gastrointestinal/genética , Metabolismo dos Lipídeos , Masculino , Salmo salar/genética , Aumento de Peso
5.
J Forensic Sci ; 65(3): 960-965, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31688972

RESUMO

In unison, fingerprinting and DNA analysis have played a pivotal role in forensic investigations. Fingerprint powders that are available on the market can come in a range of colors and with specific properties. This study evaluated the efficiency of DNA extraction from samples coated with 3 brands of fingerprint powders: Lightning, Sirchie, and SupraNano, covering a range of colors and properties. A total of 23 fingerprint powders were tested using the Chelex, Promega DNA IQ™, and Applied Biosystems™ PrepFiler™ DNA extraction protocols. The DNA IQ™ and PrepFiler™ methods extracted higher yields of DNA in comparison to Chelex, which also accounted for better quality of PowerPlex x00AE; 21 DNA profiles recovered. There were no signs of degradation or inhibition in the quantification data, indicating that samples returning low DNA yield was due to interference during DNA extraction and not PCR inhibition. DNA profiles were recovered from the majority of fingerprint powders with only a single powder, Sirchie Magnetic Silver, failing to produce a profile using any of the methods tested. A link was observed between the DNA extraction chemistry, fingerprint powder property, that is, nonmagnetic, magnetic and aqueous, and the brand of fingerprint powder. Overall, the DNA IQ™ method was favorable for nonmagnetic fingerprint powders, while magnetic fingerprint powders produced more DNA profiles when extracted with the PrepFiler™ chemistry. This study highlights the importance of screening DNA extraction chemistries for the type of fingerprint powder used, as there is not a single DNA extraction method that suits all fingerprint powder brands and properties.


Assuntos
Impressões Digitais de DNA/métodos , DNA/isolamento & purificação , Dermatoglifia , Pós/química , Humanos , Reação em Cadeia da Polimerase
6.
Chem Commun (Camb) ; 54(36): 4593-4596, 2018 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-29670956

RESUMO

We have developed a novel functional nucleic acid aptamer to amyloid-ß peptide 1-40 (Aß1-40) and investigated its potential to detect Aß peptide fragments in neuropathologically confirmed Alzheimer brain hippocampus tissues samples. Our results demonstrate that the aptamer candidate RNV95 could detect tetrameric/pentameric low-molecular-weight Aß aggregates in autopsy hippocampal tissue from two neuropathologically confirmed Alzheimer disease cases. Although these are preliminary observations, detailed investigations are under way. This is the first demonstration of aptamer-Aß binding in Alzheimer brain tissues.


Assuntos
Doença de Alzheimer/diagnóstico por imagem , Peptídeos beta-Amiloides/metabolismo , Amiloide/metabolismo , Aptâmeros de Nucleotídeos/metabolismo , Hipocampo/patologia , Fragmentos de Peptídeos/metabolismo , Doença de Alzheimer/patologia , Amiloide/química , Peptídeos beta-Amiloides/química , Aptâmeros de Nucleotídeos/química , Humanos , Fragmentos de Peptídeos/química , Placa Amiloide/diagnóstico por imagem , Placa Amiloide/patologia , Agregação Patológica de Proteínas/metabolismo , Ligação Proteica , Multimerização Proteica
7.
Mucosal Immunol ; 11(3): 774-784, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29210363

RESUMO

The intestinal epithelial cells constitute the first line of defense against gut microbes, which includes secretion of various antimicrobial substances. Reactive oxygen species (ROS) are well characterized as part of the innate phagocytic immunity; however, a role in controlling microorganisms in the gut lumen is less clear. Here, we show a role for nitric oxide synthase (iNOS)- and NOX1-produced ROS in maintaining homeostasis of the gut microbiota. In vivo imaging revealed distinctly high levels of ROS in the ileum of normal healthy mice, regulated in accordance with the amount of gut bacteria. The ROS level was dependent on the nitric oxide and superoxide producers iNOS and NOX1, respectively, suggesting peroxynitrite as the effector molecule. In the ileum of iNOS- and NOX1-deficient mice, the bacterial load is increased and the composition is more cecum like. Our data suggest a unique role of ileum in maintaining homeostasis of gut microbes through production of ROS with potential importance for preventing reflux from the large intestine, bacterial overgrowth, and translocation.


Assuntos
Bactérias/crescimento & desenvolvimento , Ceco/microbiologia , Microbioma Gastrointestinal/fisiologia , Íleo/microbiologia , Mucosa Intestinal/imunologia , NADPH Oxidase 1/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Animais , Carga Bacteriana , Ceco/imunologia , Homeostase , Íleo/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NADPH Oxidase 1/genética , Óxido Nítrico Sintase Tipo II/genética , Ácido Peroxinitroso/metabolismo , Espécies Reativas de Oxigênio/metabolismo
8.
Lett Appl Microbiol ; 65(1): 66-72, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28418627

RESUMO

Biological treatment represents a key step in nutrient removal from wastewater. Until now these process has mainly been considered prokaryotic, with the interactions between prokaryotes and eukaryotes not being properly explored. We therefore investigated the co-occurrence of eukaryotes and prokaryotes in biological nitrogen removal biofilms. We found that biofilms in the nitrifying reactor contained the highest diversity and abundance of both prokaryotes and eukaryotes, with nearly three times higher prokaryote species richness than for the denitrifying reactor. The positive associations between eukaryote abundance and prokaryote diversity could potentially be explained by mutualism - and/or predator/prey interactions. Further mechanistic insight, however, is needed to determine the main diversifying mechanisms. In summary, eukaryote and prokaryote interactions seem to play a fundamental yet underexplored role in biological wastewater treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: Eukaryote and prokaryote interactions may play an important role in wastewater treatment. This study found that prokaryote species richness was nearly three times higher in the aerobe nitrification than in an anaerobe denitrification reactor, coinciding with the highest level of eukaryotes. This knowledge can be important in process control, and potentially in the development of novel approaches based on nitrate accumulating denitrifying eukaryotes.


Assuntos
Biofilmes/crescimento & desenvolvimento , Oligoquetos/crescimento & desenvolvimento , Proteobactérias/isolamento & purificação , Rhizaria/isolamento & purificação , Águas Residuárias/microbiologia , Águas Residuárias/parasitologia , Animais , Reatores Biológicos/microbiologia , Reatores Biológicos/parasitologia , Nitrificação/fisiologia , Nitrogênio/metabolismo
9.
Environ Microbiol ; 18(7): 2226-36, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26913851

RESUMO

Transition from an infant to an adult associated gut microbiota with age through establishment of strict anaerobic bacteria remains one of the key unresolved questions in gut microbial ecology. Here a comprehensive comparative analysis of stool microbiota in a large cohort of mothers and their children sampled longitudinally up until 2 years of age using sequencing analysis tool was presented that allows realistic microbial diversity estimates. In this work, evidence for the switch from children to adult associated microbial profile between 1 and 2 years of age was provided, suggestively driven by Bifidobacterium breve. An Operational Taxonomic Unit (OTU) belonging to B. breve was highly prevalent in the population throughout the first year of life, and was negatively associated with detection of a range of adult-like OTUs. Although an adult profile was not fully established by 2 years of age, it was demonstrated that with regards to the most prevalent OTUs, their prevalence in the child population by then already resembled that of the adult population. Taken together, it was proposed that late-colonizing OTUs were recruited at a later stage and were not acquired at birth with the recruitment being controlled by gatekeeping OTUs until the age of 1 year.


Assuntos
Bactérias/isolamento & purificação , Microbioma Gastrointestinal , Adulto , Fatores Etários , Bactérias/classificação , Bactérias/genética , Pré-Escolar , Fezes/microbiologia , Feminino , Humanos , Lactente , Masculino , Filogenia , Adulto Jovem
10.
Benef Microbes ; 6(5): 657-9, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26036144

RESUMO

A key message from a range of high profile next generation sequencing studies on the human microbiota is that it composes a tremendously rich community of more than 1000 species within each one of us. Although more recent studies have shown estimates of between 100 and 200 species per individual, this has not yet been made clear in the literature. Currently, the most widely accepted estimate of species richness is therefore five to ten times too high. Here, we will review the different estimates of species richness in the literature, address potential sources of artefacts, the reluctance to correct these, and provide suggestions for future directions.


Assuntos
Biodiversidade , Microbioma Gastrointestinal , Microbiota , DNA Ribossômico/química , DNA Ribossômico/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , RNA Ribossômico 16S/genética
11.
Aliment Pharmacol Ther ; 42(1): 71-83, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25973666

RESUMO

BACKGROUND: Dysbiosis is associated with many diseases, including irritable bowel syndrome (IBS), inflammatory bowel diseases (IBD), obesity and diabetes. Potential clinical impact of imbalance in the intestinal microbiota suggests need for new standardised diagnostic methods to facilitate microbiome profiling. AIM: To develop and validate a novel diagnostic test using faecal samples to profile the intestinal microbiota and identify and characterise dysbiosis. METHODS: Fifty-four DNA probes targeting ≥300 bacteria on different taxonomic levels were selected based on ability to distinguish between healthy controls and IBS patients in faecal samples. Overall, 165 healthy controls (normobiotic reference collection) were used to develop a dysbiosis model with a bacterial profile and Dysbiosis Index score output. The model algorithmically assesses faecal bacterial abundance and profile, and potential clinically relevant deviation in the microbiome from normobiosis. This model was tested in different samples from healthy volunteers and IBS and IBD patients (n = 330) to determine the ability to detect dysbiosis. RESULTS: Validation confirms dysbiosis was detected in 73% of IBS patients, 70% of treatment-naïve IBD patients and 80% of IBD patients in remission, vs. 16% of healthy individuals. Comparison of deep sequencing and the GA-map Dysbiosis Test, (Genetic Analysis AS, Oslo, Norway) illustrated good agreement in bacterial capture; the latter showing higher resolution by targeting pre-determined highly relevant bacteria. CONCLUSIONS: The GA-map Dysbiosis Test identifies and characterises dysbiosis in IBS and IBD patients, and provides insight into a patient's intestinal microbiota. Evaluating microbiota as a diagnostic strategy may allow monitoring of prescribed treatment regimens and improvement in new therapeutic approaches.


Assuntos
Disbiose/diagnóstico , Microbioma Gastrointestinal , Doenças Inflamatórias Intestinais/microbiologia , Síndrome do Intestino Irritável/microbiologia , Adolescente , Adulto , Idoso , Bactérias/isolamento & purificação , Testes Diagnósticos de Rotina/métodos , Fezes/microbiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Noruega , Adulto Jovem
12.
J Alzheimers Dis ; 42(4): 1443-60, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25024349

RESUMO

BACKGROUND: Imaging of human brain as well as cellular and animal models has highlighted a role for the actin cytoskeleton in the development of cell pathology in Alzheimer's disease (AD). Rods and aggregates of the actin-associated protein cofilin are abundant in grey matter of postmortem AD brain and rods are found inside neurites in animal and cell models of AD. OBJECTIVE: We sought further understanding of the significance of cofilin rods/aggregates to the disease process: Do rods/aggregates correlate with AD progression and the development of hallmark neurofibrillary tangles and neuropil threads? Are cofilin rods/aggregates found in the same neurites as hyperphosphorylated tau? METHODS: The specificity of rods/aggregates to AD compared with general aging and their spatial relationship to tau protein was examined in postmortem human hippocampus, inferior temporal cortex, and anterior cingulate cortex. RESULTS: The presence of cofilin rods/aggregates correlated with the extent of tau pathology independent of patient age. Densities of rods/aggregates were fourfold greater in AD compared with aged-matched control brains and rods/aggregates were significantly larger in AD brain. We did not find evidence for our hypothesis that intracellular cofilin rods are localized to tau-positive neuropil threads. Instead, data suggest the involvement of microglia in the clearance of cofilin rods/aggregates and/or in their synthesis in and around amyloid plaques and surrounding neuropil. CONCLUSION: Cofilin rods and aggregates signify events initiated early in the pathological cascade. Further definition of the mechanisms leading to their formation in the human brain will provide insights into the cellular causes of AD.


Assuntos
Fatores de Despolimerização de Actina/metabolismo , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Encéfalo/metabolismo , Encéfalo/patologia , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/metabolismo , Envelhecimento/patologia , Western Blotting , Encéfalo/irrigação sanguínea , Feminino , Imunofluorescência , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Imageamento Tridimensional , Proteínas de Filamentos Intermediários/metabolismo , Masculino , Microglia/metabolismo , Microglia/patologia , Microscopia Confocal , Pessoa de Meia-Idade , Proteínas tau/metabolismo
13.
Neurogastroenterol Motil ; 26(8): 1155-62, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24888394

RESUMO

BACKGROUND: Depression is a chronic syndrome with a pathogenesis linked to various genetic, biological, and environmental factors. Several links between gut microbiota and depression have been established in animal models. In humans, however, few correlations have yet been demonstrated. The aim of our work was therefore to identify potential correlations between human fecal microbiota (as a proxy for gut microbiota) and depression. METHODS: We analyzed fecal samples from 55 people, 37 patients, and 18 non-depressed controls. Our analyses were based on data generated by Illumina deep sequencing of 16S rRNA gene amplicons. KEY RESULTS: We found several correlations between depression and fecal microbiota. The correlations, however, showed opposite directions even for closely related Operational Taxonomic Units (OTU's), but were still associated with certain higher order phylogroups. The order Bacteroidales showed an overrepresentation (p = 0.05), while the family Lachnospiraceae showed an underrepresentation (p = 0.02) of OTU's associated with depression. At low taxonomic levels, there was one clade consisting of five OTU's within the genus Oscillibacter, and one clade within Alistipes (consisting of four OTU's) that showed a significant association with depression (p = 0.03 and 0.01, respectively). CONCLUSIONS & INFERENCES: The Oscillibacter type strain has valeric acid as its main metabolic end product, a homolog of neurotransmitter GABA, while Alistipes has previously been shown to be associated with induced stress in mice. In conclusion, the taxonomic correlations detected here may therefore correspond to mechanistic models.


Assuntos
Transtorno Depressivo/microbiologia , Fezes/microbiologia , Microbiota , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , RNA Ribossômico 16S/genética
14.
Benef Microbes ; 4(3): 219-22, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23887030

RESUMO

The current leading view is that functionality and not phylotype is the most important determinant for the services provided by the gut microbiota. Here we present an alternative opinion, advocating the importance of phylotype in addition to function. We believe the literature is misled by technical artifacts in defining operational taxonomic units (OTUs), which are binned groups of bacteria based on sequence homology. Furthermore, the current metagenomic approaches where the total DNA in a sample is mixed prior to sequencing and subsequently resolved by a bioinformatics approach, are highly error prone with respect to both functional and phylotype assignments. We argue that the directions of the OTU and metagenome errors are such that stable phylotypes are overlooked, while functional stability is overestimated. Taking these errors into account, we propose that phylotype represents an interface for functionality, and is for this reason an important determinant for the services provided by the gut microbiota to the host.


Assuntos
Biota , Trato Gastrointestinal/microbiologia , Estilo de Vida , Humanos , Metagenômica/métodos
15.
Curr Alzheimer Res ; 10(4): 442-8, 2013 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-23270374

RESUMO

Deposition of amyloid-ß (Aß) peptides in the brain is a central event in the pathogenesis of Alzheimer's disease (AD), which makes Aß peptides a crucial target for therapeutic intervention. Significant efforts have been made towards the development of ligands that bind to Aß peptides with a goal of early detection of amyloid aggregation and the neutralization of Aß toxicity. Short single-stranded oligonucleotide aptamers bind with high affinity and specificity to their targets. Aptamers that specifically bind to Aß monomers, specifically the 40 and 42 amino acid species (Aß(1-40) and Aß(1- 42)), fibrils and plaques have a great potential for diagnostic applications and the treatment of AD. Herein, we review the aptamers that bind to the various forms of Aß peptides for use in diagnosis and to inhibit plaque formation.


Assuntos
Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/metabolismo , Aptâmeros de Nucleotídeos/uso terapêutico , Encéfalo , Doença de Alzheimer/diagnóstico , Peptídeos beta-Amiloides/química , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Encéfalo/patologia , Humanos
16.
J Appl Microbiol ; 114(1): 141-51, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23039191

RESUMO

AIMS: A major challenge in metagenome studies is to estimate the true size of all combined genomes. Here, we present a novel approach to estimate the size of all combined genomes for low coverage next-generation sequencing (NGS) data through empirically determined copy numbers of random DNA fragments. METHODS AND RESULTS: Size estimates were made based on analyses of two experimental soil micro-ecosystems - simulating soil with and without earthworms. Our analyses showed combined genome sizes of about log 11 nucleotides for each of the soil micro-ecosystems, as estimated from qPCR determined copy numbers of random DNA fragments. This corresponds to more than 20000 unique bacterial genomes in each sample. There seemed, however, to be a bacterial subpopulation in the earthworm soil, not being present in the nonearthworm soil. To describe the structure of the metagenomes, both total DNA and amplified 16S rRNA gene sequence libraries were generated with 454-sequencing. Bioinformatic analysis of 454 sequence libraries showed a large functional but low taxonomic overlap between the samples with and without earthworms. A neutrality test indicated that rare species have a competitive advantage over abundant species in both micro-ecosystems providing a potential explanation for the large metagenome sizes. CONCLUSIONS: We have shown that the soil metagenome is very large and that the large size is probably a consequence of top-down selection of the dominant bacterial species. SIGNIFICANCE AND IMPACT OF THE STUDY: Estimates of metagenome size from low coverage NGS data will be important for guiding future NGS set-ups.


Assuntos
Bactérias/classificação , Ecossistema , Metagenoma , Microbiologia do Solo , Animais , Bactérias/genética , DNA Bacteriano/genética , Genoma Bacteriano , Biblioteca Genômica , Oligoquetos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA/métodos , Solo/análise
17.
Appl Environ Microbiol ; 79(2): 497-507, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23124244

RESUMO

Bifidobacteria are a major microbial component of infant gut microbiota, which is believed to promote health benefits for the host and stimulate maturation of the immune system. Despite their perceived importance, very little is known about the natural development of and possible correlations between bifidobacteria in human populations. To address this knowledge gap, we analyzed stool samples from a randomly selected healthy cohort of 87 infants and their mothers with >90% of vaginal delivery and nearly 100% breast-feeding at 4 months. Fecal material was sampled during pregnancy, at 3 and 10 days, at 4 months, and at 1 and 2 years after birth. Stool samples were predicted to be rich in the species Bifidobacterium adolescentis, B. bifidum, B. dentium, B. breve, and B. longum. Due to high variation, we did not identify a clear age-related structure at the individual level. Within the population as a whole, however, there were clear age-related successions. Negative correlations between the B. longum group and B. adolescentis were detected in adults and in 1- and 2-year-old children, whereas negative correlations between B. longum and B. breve were characteristic for newborns and 4-month-old infants. The highly structured age-related development of and correlation networks between bifidobacterial species during the first 2 years of life mirrors their different or competing nutritional requirements, which in turn may be associated with specific biological functions in the development of healthy gut.


Assuntos
Bifidobacterium/classificação , Bifidobacterium/isolamento & purificação , Biodiversidade , Trato Gastrointestinal/microbiologia , Variação Genética , Criança , DNA Bacteriano/química , DNA Bacteriano/genética , Fezes/microbiologia , Humanos , Estudos Longitudinais , Dados de Sequência Molecular , Mães , Análise de Sequência de DNA
18.
J Dairy Sci ; 95(9): 4804-4812, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22916884

RESUMO

The nonstarter lactic acid bacteria (NSLAB) constitute an important microbial group found during cheese ripening and they are thought to be fundamental to the quality of cheese. Rapid and accurate diagnostic tests for NSLAB are important for cheese quality control and in understanding the cheese ripening process. Here, we present a novel rapid approach for strain-level characterization through combined 16S rRNA gene and repetitive sequence-based high-resolution melt analysis (HRM). The approach was demonstrated through the characterization of 94 isolates from Norvegia, a Gouda-type cheese. The HRM profiles of the V1 and V3 variable regions of the 16S rRNA gene of the isolates were compared with the HRM profiles of 13 reference strains. The HRM profile comparison of the V1 and V3 regions of the 16S rRNA gene allowed discrimination of isolates and reference strains. Among the cheese isolates, Lactobacillus casei/paracasei (62 isolates) and Lactobacillus plantarum/Lactobacillus pentosus (27 isolates) were the dominant species, whereas Lactobacillus curvatus/Lactobacillus sakei were found occasionally (5 isolates). The HRM profiling of repetitive sequence-based PCR using the (GTG)(5) primer was developed for strain-level characterization. The clustering analysis of the HRM profiles showed high discriminatory power, similar to that of cluster analysis based on the gel method. In conclusion, the HRM approach in this study may be applied as a fast, accurate, and reproducible method for characterization of the NSLAB microflora in cheese and may be applicable to other microbial environments following selective plate culturing.


Assuntos
Queijo/microbiologia , Lactobacillus/metabolismo , Impressões Digitais de DNA , Manipulação de Alimentos , Lactobacillus/genética , Lacticaseibacillus casei/genética , Lacticaseibacillus casei/metabolismo , Lactobacillus plantarum/genética , Lactobacillus plantarum/metabolismo , Desnaturação de Ácido Nucleico , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Análise de Sequência de RNA
19.
J Appl Microbiol ; 113(5): 1176-83, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22817452

RESUMO

AIM: We have tested the effect of feed structure and feeding regime to prevent the spread of the zoonotic pathogen Campylobacter jejuni in broiler chicken flocks. METHODS AND RESULTS: Birds were offered two types of feed, control diet and a diet supplemented with 15% oat/barley hulls for structure. In addition, the birds were either fed ad libitum or intermittent. One bird in each treatment group was infected with a three-strain-mix of Camp. jejuni, and the spread of Camp. jejuni within the group was investigated. Feed structure increased the gizzard weight, delayed the spread of Camp. jejuni within the group and reduced the relative amount of Camp. jejuni in the caecum compared with the control diet. CONCLUSIONS: Our results show that stimulating the bird's natural barriers is a novel and promising intervention strategy to reduce the spread of Camp. jejuni in chicken flocks. SIGNIFICANCE AND IMPACT OF THE STUDY: Preventing Camp. jejuni in broiler chicken flocks is essential to ensure food safety because this bacterium is transferred to chicken carcasses during the slaughter process and readily survive in unprocessed poultry products. We have evaluated a novel approach for stimulation of the bird's natural barriers in the upper digestive tract with promising results.


Assuntos
Ração Animal , Infecções por Campylobacter/veterinária , Campylobacter jejuni/crescimento & desenvolvimento , Galinhas/microbiologia , Doenças das Aves Domésticas/prevenção & controle , Criação de Animais Domésticos/métodos , Animais , Infecções por Campylobacter/prevenção & controle , Campylobacter jejuni/genética , Campylobacter jejuni/isolamento & purificação , Galinhas/fisiologia , DNA Bacteriano/isolamento & purificação , Dieta/veterinária , Moela das Aves/fisiologia , Masculino , Tamanho do Órgão , Doenças das Aves Domésticas/microbiologia
20.
Lett Appl Microbiol ; 54(5): 447-54, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22385401

RESUMO

AIMS: The impact of bacterial transmission from mother to child on human allergy development is poorly understood. The aim of the present work was therefore to use a temporal collected dataset of 117 mothers and their children to model the potential effect of mother-to-child bacterial transmission on allergy (IgE) sensitization. METHODS AND RESULTS: We have recently shown a negative IgE correlation to high Escherichia coli levels until the age of 1 year, with a shift to positive correlation to high Bacteroides fragilis levels at the age of 2. In the present work, we used the previous published data to model the persistence and interaction effects of E. coli and B. fragilis with respect to IgE sensitization. Temporal modelling was made by first defining a stochastic model for sensitization state based on Markov chains and regression tree analyses. Subsequent simulations were used to determine the impact of mother-to-infant bacterial transmission. The regression tree analyses showed that E. coli colonization within 4 days was negatively correlated to sensitization, while lack of E. coli colonization at day 4 combined with B. fragilis colonization after 4 months was positively correlated. With Markov chain analyses, we found that E. coli was highly persistent in infants until the age of 4 months, while the persistence of B. fragilis increased with age. CONCLUSIONS: Simulations showed that the mother's bacterial composition correlated significantly to the child's IgE sensitization state at the age of 2 years. High E. coli and low B. fragilis levels in the mother were negatively correlated, while low E. coli and high B. fragilis were positively correlated to IgE. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results support that allergy could partly be communicable, being transferred from mother to infant through the gut microbiota.


Assuntos
Alérgenos/imunologia , Bactérias/crescimento & desenvolvimento , Hipersensibilidade Imediata/imunologia , Imunoglobulina E/imunologia , Modelos Biológicos , Adulto , Bactérias/isolamento & purificação , Bacteroides fragilis/crescimento & desenvolvimento , Bacteroides fragilis/isolamento & purificação , Pré-Escolar , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/isolamento & purificação , Feminino , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Humanos , Hipersensibilidade Imediata/epidemiologia , Hipersensibilidade Imediata/metabolismo , Hipersensibilidade Imediata/microbiologia , Imunoglobulina E/metabolismo , Lactente , Recém-Nascido , Transmissão Vertical de Doenças Infecciosas , Masculino
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