RESUMO
Ruminant animals have a symbiotic relationship with the microorganisms in their rumens. In this relationship, rumen microbes efficiently degrade complex plant-derived compounds into smaller digestible compounds, a process that is very likely associated with host animal feed efficiency. The resulting simpler metabolites can then be absorbed by the host and converted into other compounds by host enzymes. We used a microbial community metabolic network inferred from shotgun metagenomics data to assess how this metabolic system differs between animals that are able to turn ingested feedstuffs into body mass with high efficiency and those that are not. We conducted shotgun sequencing of microbial DNA from the rumen contents of 16 sheep that differed in their residual feed intake (RFI), a measure of feed efficiency. Metagenomic reads from each sheep were mapped onto a database-derived microbial metabolic network, which was linked to the sheep metabolic network by interface metabolites (metabolites transferred from microbes to host). No single enzyme was identified as being significantly different in abundance between the low and high RFI animals (P > 0.05, Wilcoxon test). However, when we analyzed the metabolic network as a whole, we found several differences between efficient and inefficient animals. Microbes from low RFI (efficient) animals use a suite of enzymes closer in network space to the host's reactions than those of the high RFI (inefficient) animals. Similarly, low RFI animals have microbial metabolic networks that, on average, contain reactions using shorter carbon chains than do those of high RFI animals, potentially allowing the host animals to extract metabolites more efficiently. Finally, the efficient animals possess community networks with greater Shannon diversity among their enzymes than do inefficient ones. Thus, our system approach to the ruminal microbiome identified differences attributable to feed efficiency in the structure of the microbes' community metabolic network that were undetected at the level of individual microbial taxa or reactions.
Assuntos
Ração Animal/análise , Microbioma Gastrointestinal , Redes e Vias Metabólicas , Metagenômica , Ovinos/fisiologia , Animais , Feminino , Rúmen/metabolismo , Rúmen/microbiologia , Ovinos/microbiologiaRESUMO
BACKGROUND: Grazing mammals rely on their ruminal microbial symbionts to convert plant structural biomass into metabolites they can assimilate. To explore how this complex metabolic system adapts to the host animal's diet, we inferred a microbiome-level metabolic network from shotgun metagenomic data. RESULTS: Using comparative genomics, we then linked this microbial network to that of the host animal using a set of interface metabolites likely to be transferred to the host. When the host sheep were fed a grain-based diet, the induced microbial metabolic network showed several critical differences from those seen on the evolved forage-based diet. Grain-based (e.g., concentrate) diets tend to be dominated by a smaller set of reactions that employ metabolites that are nearer in network space to the host's metabolism. In addition, these reactions are more central in the network and employ substrates with shorter carbon backbones. Despite this apparent lower complexity, the concentrate-associated metabolic networks are actually more dissimilar from each other than are those of forage-fed animals. Because both groups of animals were initially fed on a forage diet, we propose that the diet switch drove the appearance of a number of different microbial networks, including a degenerate network characterized by an inefficient use of dietary nutrients. We used network simulations to show that such disparate networks are not an unexpected result of a diet shift. CONCLUSION: We argue that network approaches, particularly those that link the microbial network with that of the host, illuminate aspects of the structure of the microbiome not seen from a strictly taxonomic perspective. In particular, different diets induce predictable and significant differences in the enzymes used by the microbiome. Nonetheless, there are clearly a number of microbiomes of differing structure that show similar functional properties. Changes such as a diet shift uncover more of this type of diversity.
Assuntos
Dieta , Microbioma Gastrointestinal/fisiologia , Redes e Vias Metabólicas , Metagenômica , Rúmen/microbiologia , Ovinos/microbiologia , Ração Animal/análise , Animais , Digestão/fisiologia , Grão Comestível , Comportamento Alimentar , Rúmen/fisiologia , Ovinos/fisiologiaRESUMO
This study explored the link between vitamin D status and frequency of skin infections, inflammation, and injury in college wrestlers during an academic year. METHODS: Serum 25-hydroxyvitamin D (25(OH)D) (n = 19), plasma cytokine (TNF-α, IL-6, IL-10) (n = 18) concentrations, and body weight/composition were measured and injury/illness/skin infection data were collected in fall, winter, and spring. RESULTS: In the fall, 74% of wrestlers had vitamin D concentrations <32 ng/mL which increased to 94% in winter and spring. Wrestlers lost an average of 3.4 ± 3.9 kg (p < 0.001) during the season with corresponding decreases in fat mass and increases in lean mass (p < 0.01). An inverse association between 25(OH)D concentrations and total body mass and body fat percentage was observed at all-time points (p < 0.01). Concentrations of cytokines were highly variable among individuals and did not change across time (p > 0.05). Correlations between vitamin D status, cytokines, or frequency of illness, injury, or skin infections were not observed. CONCLUSIONS: A high prevalence of vitamin D insufficiency (<32 ng/mL) and deficiency (<20 ng/mL) was observed in wrestlers and was associated with higher adiposity. It remains unclear if higher vitamin D status would reduce injury, illness, and skin infection risk.
Assuntos
Peso Corporal , Inflamação/sangue , Vitamina D/análogos & derivados , Vitamina D/sangue , Adiposidade , Adolescente , Atletas , Citocinas/sangue , Citocinas/genética , Citocinas/metabolismo , Regulação da Expressão Gênica , Humanos , Inflamação/metabolismo , Masculino , Estações do Ano , Esportes/fisiologia , Deficiência de Vitamina D/sangue , Luta Romana , Adulto JovemRESUMO
An early response of the human and bovine endometrium to pregnancy is induction of an interferon (IFN)-stimulated gene (ISG) that encodes the ubiquitin-related protein, ISG15. Because the mode of implantation differs among species, we tested whether Isg15 mRNA was also expressed in the mouse uterus in response to the implanting conceptus. Isg15 mRNA was detected in the mouse uterus and increased after d 4.5 of pregnancy but did not change between d 3.5 and 9.5 of pseudopregnancy. Within the decidua, Isg15 mRNA was localized to the antimesometrial zone of the implantation sites. The level of Isg15 mRNA in artificially induced deciduomas was similar to the nonpregnant uterus and was approximately 10-fold lower than in the pregnant uterus. In vitro, murine decidual cells derived from artificially induced deciduomas could be induced to produce the Isg15 protein as well as Isg15-conjugated proteins when stimulated with type 1 IFN, though were less responsive to IFN-gamma. Isg15 is one of few gene products identified in murine implantation sites to require presence of the conceptus and not simply differentiation of the stroma. In vitro data support the inference that the pregnancy-specific inducer of uterine Isg15 is a type 1 IFN or a cytokine that signals through a similar pathway.
Assuntos
Citocinas/genética , Implantação do Embrião/fisiologia , Útero/fisiologia , Animais , Decídua/fisiologia , Feminino , Expressão Gênica/fisiologia , Hibridização In Situ , Masculino , Camundongos , Camundongos Endogâmicos ICR , Gravidez , Pseudogravidez/fisiopatologia , RNA Mensageiro/análise , Ubiquitinas , Regulação para Cima/fisiologiaRESUMO
The interferon-stimulated gene ISG15, a ubiquitin homolog, becomes conjugated to and regulates uterine proteins in response to conceptus-derived interferon-tau on d 18 of pregnancy. It was hypothesized here that cellular localization of ISG15 within endometrial cells might provide insight regarding function. Uteri were collected from cows (approximately 21-d estrous cycle) on d 17-21/0 of the estrous cycle and pregnancy and d 23, 45, and 50 of pregnancy. Intracellular ISG15 and its conjugates were present on d 17 of pregnancy, peaked to highest levels from d 18 to 23 and then declined to low but detectable levels by d 45 (P < 0.05) based on Western blotting. ISG15 and its conjugates were not detected on d 50 of pregnancy or during the estrous cycle. Immunohistochemistry revealed that ISG15 was localized throughout the endometrium on d 18-23, with heaviest staining in the sublumenal stratum compactum and the glandular epithelium throughout the stratum spongiosum. By d 45 and 50, ISG15 was lightly stained only in the stratum compactum immediately beneath the lumenal epithelium. Using transmission electron microscopy and immunogold labeling, ISG15 was specifically localized to organelles and compartments of endometrial epithelial cells and stromal cells: nucleus, perinuclear space, cytosol, mitochondria, rough endoplasmic reticulum, and cell membrane. This specific localization in epithelial and stromal cells led to the conclusion that ISG15 has diverse intracellular functions. The sustained presence of conjugated ISG15 through d 50 of pregnancy might reflect stabilization of conjugated proteins in response to implantation and the development of the placenta.
Assuntos
Citocinas/análise , Endométrio/química , Imuno-Histoquímica , Microscopia Eletrônica , Animais , Anticorpos Monoclonais , Western Blotting , Bovinos , Membrana Celular/química , Núcleo Celular/química , Citosol/química , Endométrio/ultraestrutura , Retículo Endoplasmático Rugoso/química , Células Epiteliais/ultraestrutura , Feminino , Idade Gestacional , Mitocôndrias/química , Organelas/química , Gravidez , Células Estromais/ultraestrutura , Distribuição Tecidual , Ubiquitina/análise , Ubiquitina/metabolismoRESUMO
We surveyed the ruminal metagenomes of 16 sheep under two different diets using Illumina pair-end DNA sequencing of raw microbial DNA extracted from rumen samples. The resulting sequence data were bioinformatically mapped to known prokaryotic 16S rDNA sequences to identify the taxa present in the samples and then analysed for the presence of potentially new taxa. Strikingly, the majority of the microbial individuals found did not map to known taxa from 16S sequence databases. We used a novel statistical modelling approach to compare the taxonomic distributions between animals fed a forage-based diet and those fed concentrated grains. With this model, we found significant differences between the two groups both in the dominant taxa present in the rumen and in the overall shape of the taxa abundance curves. In general, forage-fed animals have a more diverse microbial ecosystem, whereas the concentrate-fed animals have ruminal systems more heavily dominated by a few taxa. As expected, organisms from methanogenic groups are more prevalent in forage-fed animals. Finally, all of these differences appear to be grounded in an underlying common input of new microbial individuals into the rumen environment, with common organisms from one feed group being present in the other, but at much lower abundance.
Assuntos
Bactérias/genética , Dieta , Metagenoma , Rúmen/microbiologia , Ovinos/microbiologia , Animais , Bactérias/classificação , DNA Ribossômico/genética , Ecossistema , Análise de Sequência de DNARESUMO
Sexual differentiation of the brain occurs between d 30 and 70 in the fetal lamb. The objective of this experiment was to determine if maternal fatness affects fetal steroid production and expression of their receptors which may ultimately alter endocrine systems postnatally. Fetuses were collected from ewes fed at either 100% (Control; n=5) or 150% (Fat; n=6) of NRC recommendations from 60 d prior to breeding until collection at 75 d of gestation. Hypothalamic and amygdala neural tissues were collected from twin male/female fetuses. Serum concentrations of testosterone were greater (P<0.001) in male fetuses compared to female fetuses. Further, male fetuses from Fat ewes had greater (P<0.05) serum concentrations of testosterone than male fetuses from Control ewes, but differences in testicular steroidogenic enzyme mRNA were not detected (P=0.18). Quantity of hypothalamic mRNA for estrogen receptor (ER) beta tended (P=0.1) to be influenced by a sex by treatment interaction. Messenger RNA for ER-beta was greater in female fetuses than male fetuses from Control ewes (P=0.05). Although amount of ER-beta mRNA did not differ among male fetuses (P=0.7), amounts tended to be less (P=0.07) in female fetuses from Fat ewes compared to those from Control ewes, and did not differ (P> or =0.8) from male fetuses. Hypothalamic ER-alpha mRNA tended (P=0.1) to be less in fetuses from Fat ewes compared to Control fetuses but was not influenced (P=0.3) by fetal sex or their interaction. Amount of mRNA for hypothalamic progesterone receptor tended (P=0.06) to be greater in male fetuses than female fetuses and tended to be less (P=0.06) in fetuses from Fat ewes than in Control fetuses, but did not differ by any sex by treatment interaction (P=0.6). Hypothalamic RNA for the androgen receptor did not differ by sex, dam nutritional treatment, or the interaction. Likewise, amygdala RNA for the estrogen or androgen receptor did not differ (P> or =0.3) by sex, treatment, or their interaction. Dam fatness appears to decrease the expression of progesterone receptor, ER-alpha, and decrease amount of ER-beta in the female fetuses while increasing circulating concentrations of testosterone in male fetuses. Altered expression of hypothalamic receptor genes by the uterine environment may affect adult responses to stress, sexual behavior and/or the pattern of gonadotropin release in response to gonadal steroids.