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1.
Ecotoxicol Environ Saf ; 209: 111732, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33373928

RESUMO

Fluoride, widely presented in drinking water and tea, may be detrimental or beneficial to the human health, depending on its dosages ingested. However, the relationship of different dosages of fluoride and gut microbiota is still unclear. In this work, the fermentation model using fecal samples provided by four volunteers was used to evaluate the effects of different dosages of fluoride (1, 2, 10 and 15 mg/L) on the gut microbiota in vitro. The result showed low dosages of fluoride (1 and 2 mg/L) had limited effect on the structure and functional Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of gut microbiota. Furthermore, the low dosage of fluoride could promote the growth of beneficial gut microbiota, including Faecalibacterium and Lactobacillus. Whereas, the high dosage of fluoride (10 and 15 mg/L) significantly changed the composition and functional KEGG pathway of gut microbiota. Moreover, the high dosage of fluoride could also reduce the beneficial gut microbiota, including Faecalibacterium and Phascolarctobacterium, and increase the harmful bacterium including Proteobacteria and Enterobacteriaceae. Both low and high dosages of fluoride showed limited effect on the productions of short-chain fatty acids (SCFAs). Thus, the beneficial or detrimental fluoride to gut microbiota depends on its dosages. The fluoride is expected to serve as a food additive in suitable dosage to improve human health through modulation of the gut microbiota. Moreover, more attention should be paid to toxicity of fluoride with high dosage to gut microbiota.


Assuntos
Fluoretos/toxicidade , Microbioma Gastrointestinal/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Bactérias/metabolismo , Ácidos Graxos Voláteis/análise , Ácidos Graxos Voláteis/química , Ácidos Graxos Voláteis/metabolismo , Fezes/química , Fermentação , Fluoretos/análise , Humanos , Lactobacillus/metabolismo
2.
Nutrients ; 13(1)2020 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-33374116

RESUMO

We hypothesized that the role of microbiota in breast cancer relates to its influence on gut lipid metabolism. This was tested in an in vitro model combining MCF-7 and Caco-2 cells. A total of 32 women newly diagnosed for breast cancer before any treatment and 28 healthy women provided their stools. Bacterial DNA was amplified by qPCR targeting 16s rRNA specific to Bacteroidetes and Firmicutes phyla, Lactobacillales sp., Clostridium cluster IV, Faecalibacterium prausnitzii, Clostridium cluster XIVa, Roseburia intestinalis, Blautia sp., Lactonifactor longoviformis, Bifidobacterium sp., Coriobacteriaceae, Eggertella lenta, Escherichia, and Shigella. Fecal waters (FW) were quantified for short chain fatty acids (SCFA). Caco-2 cells grown on filter inserts were incubated apically with 10% FW for 24 h, and LXR, apolipoproteins AIV, and E gene expression were estimated by real time (RT) qPCR. Then, MCF-7 cells were incubated with the whole basolateral medium for 24 h, and their viability was estimated by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) test. Regression models were used to determine the correlation between MCF-7 viability and bacteria relative abundance, Caco-2 cells lipid metabolism gene expression and stool composition, as well as microbiota composition and short chain fatty acids. Logistic regression models established disease odds ratios (OR) for MCF-7 viability and Caco-2 gene expression. The OR of MCF-7 viability was 1.05 (1.01-1.10) (OR (5th-95th), p = 0.04), while that of apo AIV gene expression was 0.63 (0.39-1.01), p = 0.055). Viability correlated with % Bifidobacterium sp. (21.18 ± 7.66, p = 0.008) and valerate (-2.849 ± 1.048, p = 0.009) (ß ± s.d.). This study suggests that microbiota interacts with intestine cell lipid metabolism. Since these metabolites can reach breast cells by systemic circulation, we hypothesized that they may influence cancer disease.


Assuntos
Neoplasias da Mama/microbiologia , Enterócitos/metabolismo , Fezes/microbiologia , Microbioma Gastrointestinal/fisiologia , Metabolismo dos Lipídeos/fisiologia , Bactérias/classificação , Bactérias/genética , Neoplasias da Mama/patologia , Células CACO-2 , Sobrevivência Celular , DNA Bacteriano/análise , Ácidos Graxos Voláteis/análise , Feminino , Expressão Gênica , Humanos , Metabolismo dos Lipídeos/genética , Células MCF-7 , Pessoa de Meia-Idade
3.
J Oleo Sci ; 69(9): 1001-1009, 2020 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-32788519

RESUMO

To determine the food potential of Paeonia ludlowii D.Y.Hong (P. ludlowii) kernel oil, in this study, we analysed the fatty acid composition and volatile components of this oil, compared the antioxidant effects of two natural antioxidants on it, and then predicted its shelf life at room temperature (25°C). The results showed that P. ludlowii kernel oil mainly contained 20 fatty acids, of which linoleic acid, oleic acid and other unsaturated fatty acid contents together made up 86.99%. The aromatic composition of the crude P. ludlowii kernel oil was analysed, and 34 aromatic compounds were obtained, including 5 lipids (2.30%), 9 alcohols (12.64%), 6 aldehydes (14.67%), 2 alkanes (1.30%), 5 acids (2.70%), 1 ketone (0.41), 2 alkenes (39.12%) and 4 other substances (26.85%). The effects of the antioxidants were ranked as follows: 0.04% tea polyphenols + crude oil > 0.04% bamboo flavonoids + crude oil > crude oil. In addition, the shelf lives at room temperature (25℃) of each kernel oil-antioxidant mixture were 200.73 d, 134.90 d and 131.61 d, respectively. Overall, these results reveal that P. ludlowii kernel oil is a potential candidate for a new high-grade edible oil, and its development has broad application prospects.


Assuntos
Ácidos Graxos Insaturados/análise , Ácidos Graxos Voláteis/análise , Qualidade dos Alimentos , Armazenamento de Alimentos , Paeonia/química , Óleos Vegetais/química , Antioxidantes , Ácidos Graxos Insaturados/química , Ácidos Graxos Voláteis/química , Ácido Linoleico/análise , Ácido Oleico/análise , Óleos Vegetais/isolamento & purificação , Temperatura , Fatores de Tempo
5.
PLoS One ; 15(8): e0236944, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32745090

RESUMO

Gut dysbiosis has been implicated in the pathophysiology of a growing number of non-communicable diseases. High through-put sequencing technologies and short chain fatty acid (SCFA) profiling enables surveying of the composition and function of the gut microbiota and provide key insights into host-microbiome interactions. However, a methodological problem with analyzing stool samples is that samples are treated and stored differently prior to submission for analysis potentially influencing the composition of the microbiota and its metabolites. In the present study, we simulated the sample acquisition of a large-scale study, in which stool samples were stored for up to two days in the fridge or at room temperature before being handed over to the hospital. To assess the influence of time and temperature on the microbial community and on SCFA composition in a controlled experimental setting, the stool samples of 10 individuals were exposed to room and fridge temperatures for 24 and 48 hours, respectively, and analyzed using 16S rRNA gene amplicon sequencing, qPCR and nuclear magnetic resonance spectroscopy. To best of our knowledge, this is the first study to investigate the influence of storage time and temperature on the absolute abundance of methanogens, and of Lactobacillus reuteri. The results indicate that values obtained for methanogens, L. reuteri and total bacteria are still representative even after storage for up to 48 hours at RT (20°C) or 4°C. The overall microbial composition and structure appeared to be influenced more by laboratory errors introduced during sample processing than by the actual effects of temperature and time. Although microbial activity was demonstrated by elevated SCFA at both 4°C and RT, SCFAs ratios were more stable over the different conditions and may be considered as long as samples are come from similar storage conditions.


Assuntos
Fezes/química , Fezes/microbiologia , Manejo de Espécimes/métodos , Adulto , Bactérias/genética , Disbiose/microbiologia , Ácidos Graxos Voláteis/análise , Feminino , Microbioma Gastrointestinal/genética , Humanos , Masculino , Microbiota , RNA Ribossômico 16S/genética , Temperatura , Fatores de Tempo
6.
PLoS One ; 15(8): e0238006, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32857814

RESUMO

This study aimed to evaluate the effects of two prebiotics in different concentrations on nutrient digestibility, fermentative products and immunological variables in adult dogs. Twenty-four adult dogs were randomly divided into six blocks according to their metabolic body weights (BW0.75); within these groups, dogs were randomized to four treatments: control without prebiotics (CO); inclusion of 0.5% prebiotic blend Yes-Golf (B1); inclusion of 1.0% galactooligosaccharide (GOS); and inclusion of 1.0% prebiotic blend Yes-Golf (B2). The experiment lasted 30 days, with 20 days adaptation and 10 days stool and blood collection. Results were analyzed for normality and means were separated by ANOVA and adjusted by the Tukey test at the significance level of 5.0%. Prebiotic supplementation had no effect on apparent digestibility coefficients (ADC), total stool production and fecal scores (p > 0.05). Prebiotics evaluated also did not alter fecal pH, nor the concentrations of ammonia, lactic acid, short chain fatty acids (SCFA) and most fecal branched chain fatty acids (BCFA) (p > 0.05). The addition of GOS decreased the concentration of iso-valeric acid (p = 0.0423). Regarding immunological variables, concentrations of fecal IgA were not influenced by the treatments. Treatments GOS and B2 increased the total number of polymorphonuclear cells, as well as the oxidative burst in relation to treatments B1 and CO (p < 0.0001). Treatment B2 improved the rate of S. aureus phagocytosis in relation to CO (p = 0.0111), and both the GOS and B2 treatments had a better index for E. coli phagocytosis than the CO treatment (p = 0.0067). In conclusion, there was indication that both prebiotics GOS and B2 at 1.0% inclusion improved the immunity of healthy dogs.


Assuntos
Colo/efeitos dos fármacos , Oligossacarídeos/farmacologia , Prebióticos , Animais , Colo/imunologia , Colo/microbiologia , Dieta/veterinária , Cães , Ácidos Graxos Voláteis/análise , Fezes/química , Microbioma Gastrointestinal/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Leucócitos/citologia , Leucócitos/efeitos dos fármacos , Leucócitos/metabolismo , Fagocitose/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Staphylococcus aureus/fisiologia
7.
Zhonghua Wei Chang Wai Ke Za Zhi ; 23(Z1): 69-76, 2020 Jul 10.
Artigo em Chinês | MEDLINE | ID: mdl-32594729

RESUMO

Objective: To examine the association between the clinical efficacy of fecal microbiota transplantation (FMT) in recipients and the choice of donor, and to observe the characteristics of intestinal flora and metabolites among different donors. Methods: A retrospective case-control study was conducted. Donor whose feces was administrated for more than 30 recipients was enrolled. Data of 20 FMT donors and corresponding recipients at Intestinal Microecology Diagnosis and Treatment Center of the Tenth People's Hospital from October 2018 to December 2019 were collected retrospectively. During follow-up, the efficacy of each recipient 8-week after FMT treatment was recorded and analyzed. Based on the efficacy of each donor, the donors were divided into three groups: high efficacy group (effective rate >60%, 10 donors), moderate efficacy group (effective rate 30%-60%, 6 donors) and low efficacy group (effective rate <30%, 4 donors). The structure of the bacterial flora and the content of fecal short-chain fatty acids in each group of donors were detected and compared among groups. Association of the efficacy of each donor group with the morbidity of complications, and association of efficacy of recipients with donors were analyzed. The evaluation indicators of FMT efficacy included objective clinical effectiveness and/or subjective effectiveness. Objective effectiveness indicated clinical cure plus clinical improvement, and subjective effectiveness indicated marked effectiveness plus medium effectiveness through questionnaire during follow-up. Results: A total of 1387 recipients were treated by 20 donors, including 749 cases of chronic constipation, 141 cases of chronic diarrhea, 107 cases of inflammatory bowel disease (IBD), 121 cases of irritable bowel syndrome (IBS), 83 cases of autism, and 186 cases of other diseases, such as radiation bowel injury, intestinal pseudo-obstruction, paralytic intestinal obstruction, functional bloating and allergic diseases. There were 829 cases, 403 cases, and 155 cases in high efficacy group, moderate efficacy group and low efficacy group respectively. Baseline data among 3 groups were not significantly different (all P> 0.05). In comparison of bacterial abundance (operational taxonomic unit, OTU) among different effective donor groups, the high efficacy group was the highest (330.68±57.28), the moderate efficacy group was the second (237.79±41.89), and the low efficacy group was the lowest (160.60±49.61), whose difference was statistically significant (F=16.910, P<0.001). In comparison of bacterial diversity (Shannon index), the high efficacy group and the moderate efficacy group were higher (2.96±0.36 and 2.67±0.54, respectively), and the low efficacy group was lower (2.09±0.55), whose difference was statistically significant (F=5.255, P=0.017). In comparison of butyric acid content among three groups, the high efficacy group had the highest [(59.20±9.00) µmol/g], followed by middle efficacy group [(46.92±9.48) µmol/g], and the low efficacy group had the lowest [(37.23±5.03) µmol/g], whose difference was statistically significant (F=10.383, P=0.001). The differences of acetic acid and propionic acid among three groups were not statistically significant (all P>0.05). A total of 418 cases developed complications (30.1%). Morbidity of complication in low efficacy group, moderate efficacy group and high efficacy group was 40.6% (63/155), 30.0% (121/403) and 28.2% (243/829) respectively, and the difference was statistically significant (χ(2)=9.568, P=0.008). The incidence of diarrhea in low efficacy group, moderate efficacy group and high efficacy group was 7.1% (11/155), 4.0% (16/403) and 2.8% (23/829) respectively, and the difference was statistically significant (χ(2)=7.239, P=0.027). Comparing the incidences of other types of complications, no statistically significant differences were found (all P>0.05). Follow up began 8 weeks after the FMT treatment. The total follow-up rate was 83.6% (1160/1387). The overall effective rate 58.3% (676/1160). Effective rates of various diseases were as follows: chronic constipation 54.3% (328/604), chronic diarrhea 88.5% (115/130), IBD 56.1% (55/98), IBS 55.1% (59/107), autism 61.6% (45/73), and other diseases 50.0% (74/148). Comparing the effective rate of three groups of donors for different diseases, there was no statistically significant difference in chronic diarrhea (P>0.05); there was a positive correlation trend in IBD, IBS and autism, but the differences were not statistically significant (all P>0.05). For chronic constipation and other diseases, high efficacy group had the highest effective rate [65.0% (243/374) and 63.2% (55/87)], followed by moderate efficacy group [49.4% (86/174) and 38.1% (16/42)], and low efficacy group had the lowest [16.1% (9/56) and 15.8% (3/19)], whose differences were significant (all P<0.05). Conclusions: Different donors have different efficacy in different diseases. Chronic constipation, radiation bowel injury, etc. need to choose donors with high efficacy. IBD, IBS and autism may also be related to the effectiveness of donors, while chronic diarrhea is not associated to the donor. The efficiency of the donor is negatively correlated to the morbidity of complications. The abundance and diversity of intestinal flora and the content of butyric acid may affect the efficacy of the donor.


Assuntos
Transtorno Autístico/terapia , Seleção do Doador , Transplante de Microbiota Fecal/métodos , Enteropatias/terapia , Butiratos/análise , Estudos de Casos e Controles , Ácidos Graxos Voláteis/análise , Fezes/química , Fezes/microbiologia , Humanos , Estudos Retrospectivos , Resultado do Tratamento
8.
J Anim Sci ; 98(5)2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32386225

RESUMO

The objectives of this research were to evaluate the effects of source and concentration of α-amylase-treated neutral detergent fiber (aNDF) from roughage on feed intake, ingestive behavior, and ruminal kinetics in beef cattle receiving high-concentrate diets. Six ruminally cannulated Nellore steers (408 ± 12 kg of body weight) were randomly assigned to a 6 × 6 Latin square design with six diets: 10% aNDF from corn silage (10CS); 20% aNDF from corn silage (20CS); or four diets containing 10% aNDF from corn silage and 10% aNDF from one of the following sources: sugarcane (SC), sugarcane bagasse (SCB), soybean hulls (SH), or low oil cottonseed hulls (LOCH). The parameters of passage and degradation kinetics were estimated based on a two-compartmental model with gamma- and exponential-distributed residence times. The nonlinear models were fitted by nonlinear least squares, and a linear mixed-effects model was fitted to all variables measured from the Latin square design that were related to intake, digestibility, digestion kinetic parameters, and residence times. Mean particle size (MPS) between roughage sources (CS, SCB, and SC) and coproducts (SH and LOCH) was affected (P < 0.05). Dry matter intake (DMI) was not affected (P > 0.05) by 20CS, SC, SH, or LOCH. Steers fed 20CS or LOCH diets had 16% and 20% greater DMI, respectively, (P < 0.05) than steers fed 10CS diet. Steers fed SCB consumed the least dry matter (DM). The SH and LOCH diets had lower MPS values (about 8.77 mm) in comparison to 20CS, SCB, and SC diets (about 13.08 mm) and, consequently, affected (P < 0.05) rumen content, ruminal in situ disappearance, nutrient digestibility, and solid fractional passage rate. Chewing time was affected (P < 0.05) by roughage sources and concentration. Lower values of distance travel inside the rumen (min/cm) were observed (P < 0.05) for the SCB and SC diets in comparison with any other diet. Except for SCB, there was no difference (P > 0.05) in rumen fill, among other treatments. Mean daily ruminal pH was not affected (P > 0.05) by 20CS, SCB, SC, and LOCH diets, and it ranged from 6.1 to 6.23. Total short-chain fatty acids concentration was affected (P < 0.05) by roughage source and concentration. Based on our results, we recommend that under Brazilian finishing diets, replacing roughage sources, except for SCB, based on aNDF concentration of the roughage in high-concentrate diets containing finely ground flint corn does not affect DMI.


Assuntos
Bovinos/fisiologia , Fibras na Dieta/análise , Ácidos Graxos Voláteis/análise , Comportamento Alimentar , Silagem/análise , Animais , Celulose , Óleo de Sementes de Algodão , Dieta/veterinária , Ingestão de Alimentos , Cinética , Masculino , Rúmen/metabolismo , Saccharum , Soja , Zea mays
9.
J Appl Microbiol ; 129(5): 1163-1172, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32392369

RESUMO

AIMS: This study evaluated the effects of exogenous lactic acid bacteria (LAB) on silage fermentation and bacterial community of reed canary grass (RCG) straw. METHODS AND RESULTS: The leaf, stem and whole crop of RCG straw were separately ensiled in small bag silos, without (control) or with inoculation of two exogenous LAB (LP, Lactobacillus plantarum; LB, Lactobacillus buchneri), and stored at ambient temperature of <20°C. Inoculation of exogenous LAB decreased (P < 0·05) bacterial alpha diversity and shifted (P < 0·05) bacterial community compositions, but did not change (P> 0·05) the relative abundance of Lactobacillus. Particularly, inoculation of LB increased (P < 0·05) acetic acid and propionic acid contents, decreased (P < 0·05) butyric acid (BA) and ammonia-N contents, separated (P < 0·05) the bacterial community in silage. However, the exogenous LAB inoculated silages were characterized by main distribution of yeasts, presence of undesirable bacterial genera such as Clostridium and high levels of BA and ammonia-N. CONCLUSION: Inoculation of exogenous LAB exerted a limited influence on the silage fermentation and bacterial community compositions of RCG straw on the Qinghai-Tibetan Plateau. SIGNIFICANCE AND IMPACT OF THE STUDY: Commercial LAB inoculants are not always efficient on enhancing silage quality and stability. Thus, an alternative additive for inhibiting undesirable microbes during storage is important to improve RCG silage quality on the Qinghai-Tibetan Plateau.


Assuntos
Lactobacillales/fisiologia , Microbiota , Phalaris/microbiologia , Silagem/microbiologia , Amônia/análise , Amônia/metabolismo , Bactérias/classificação , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Ácidos Graxos Voláteis/análise , Ácidos Graxos Voláteis/metabolismo , Fermentação , Lactobacillales/metabolismo , Phalaris/química , Silagem/análise , Tibet , Leveduras/classificação , Leveduras/crescimento & desenvolvimento , Leveduras/metabolismo
10.
Am J Clin Nutr ; 111(6): 1286-1296, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32320024

RESUMO

BACKGROUND: The low intake of dietary fiber compared to recommended amounts has been referred to as the dietary fiber gap. The addition of fiber to snack foods could favorably alter gut microbiota and help individuals meet intake recommendations. OBJECTIVES: Our objective was to examine the effect of low- and moderate-dose fiber-containing snack bars, comprising mainly chicory root inulin-type fructans (ITF), on gut microbiota in healthy adults with habitual low dietary fiber intake using 16S ribosomal RNA-based approaches. METHODS: In 2 separate 4-wk, placebo-controlled, double-blind, crossover trials, 50 healthy adults with low dietary fiber intake were randomly assigned to receive isocaloric snack bars of either moderate-dose fiber (7 g/d) or control in Trial 1 (n = 25) or low-dose fiber (3 g/d) or control in Trial 2 (n = 25), with 4-wk washout periods. Fecal microbiota composition and inferred function, fecal SCFA concentration, gastrointestinal (GI) symptoms, dietary intake, and quality of life were measured. RESULTS: Compared with the control group, the moderate-dose group showed significant differences across multiple microbial taxa, most notably an increased relative abundance of the Bifidobacterium genus from (mean ± SEM) 5.3% ± 5.9% to 18.7% ± 15.0%. With low-dose ITF, significant increases in Bifidobacterium were no longer present after correction for multiple comparisons but targeted analysis with qPCR showed a significant increase in Bifidobacterium. Predictive functional profiling identified changes in predicted function after intake of the moderate- but not the low-dose bar. Fecal SCFAs were affected by time but not treatment. There were no between-group differences in GI symptoms. Importantly, fiber intake increased significantly with the moderate- and low-dose bars. CONCLUSIONS: In healthy adults, adding 3 or 7 g ITF to snack bars increased Bifidobacterium, a beneficial member of the gut microbial community. The addition of ITF to food products could help reduce the dietary fiber gap prevalent in modern life.This trial was registered at clinicaltrials.gov as NCT03042494.


Assuntos
Chicória/química , Fibras na Dieta/metabolismo , Microbioma Gastrointestinal , Inulina/metabolismo , Extratos Vegetais/metabolismo , Adulto , Idoso , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Chicória/metabolismo , Estudos Cross-Over , Fibras na Dieta/análise , Ácidos Graxos Voláteis/análise , Ácidos Graxos Voláteis/metabolismo , Fezes/química , Fezes/microbiologia , Feminino , Humanos , Inulina/análise , Masculino , Pessoa de Meia-Idade , Extratos Vegetais/análise , Raízes de Plantas/química , Raízes de Plantas/metabolismo , Lanches , Adulto Jovem
11.
J Anim Sci ; 98(5)2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32307528

RESUMO

The aim of this study was to investigate the effects of diet complexity and l-Thr supplementation level on the growth performance, immune response, intestinal barrier function, and microbial metabolites in nursery pigs. Thirty-two weaned pigs (body weight 7.23 ± 0.48 kg) were randomly assigned to dietary treatments in a 2 × 2 factorial arrangement based on diet complexity (complex or simple) and dietary Thr content. The complex diet contained fish meal, plasma protein, and dried whey to mimic a conventional nursery diet. The simple diet was formulated with corn, wheat, and soybean meal and did not contain any animal products. l-Thr was supplemented to each diet to supply either 100% (STD Thr) or 115% (SUP Thr) of the NRC (2012) requirement for standardized ileal digestible Thr. Pigs were individually housed and fed experimental diets ad libitum for 14 d. Diet complexity, dietary Thr content, and their interactions were considered the main effects. Pigs fed the simple diet had greater (P < 0.05) plasma interleukin (IL)-10 and IL-6 concentrations compared with those fed the complex diet on days 7 and 14, respectively. Simple diet-fed pigs tended to show greater (P < 0.10) expression of genes encoding for tumor necrosis factor-α, claudin-1, and zonula occludens-1 in the jejunum compared with complex diet-fed pigs. The simple diet-fed pigs had greater (P < 0.05) concentrations of NH3-N in the jejunum digesta than did complex diet-fed pigs. The SUP Thr increased (P < 0.05) villus height and goblet cell (GC) density in villi and crypts in the jejunum and deepened (P < 0.05) crypts in the proximal colon. The SUP Thr resulted in the upregulation (P < 0.05) of occludin gene expression and a tendency toward the downregulation (P = 0.10) of IL-6 gene expression in the jejunum. Interactions (P < 0.05) between diet complexity and l-Thr supplementation level were observed in GC density in the crypt, NH3-N concentration in the jejunum, and the contents of acetate, propionate, and total volatile fatty acids in the colon. In conclusion, feeding a simple diet to nursery pigs resulted in systemic and intestinal inflammation. The SUP Thr diet did not normalize the simple diet-induced inflammation but improved gut integrity. SUP Thr seems to have greater benefits with a simple diet than with a complex diet. Therefore, SUP Thr in a simple diet could be a beneficial nutritional strategy for enhancing gut health.


Assuntos
Suplementos Nutricionais/análise , Metabolismo Energético/efeitos dos fármacos , Microbioma Gastrointestinal/fisiologia , Imunidade Inata/efeitos dos fármacos , Suínos/fisiologia , Treonina/administração & dosagem , Ração Animal/análise , Animais , Composição Corporal , Peso Corporal/efeitos dos fármacos , Dieta/veterinária , Ácidos Graxos Voláteis/análise , Microbioma Gastrointestinal/efeitos dos fármacos , Interleucina-10/análise , Mucosa Intestinal/efeitos dos fármacos , Masculino , Nitrogênio/metabolismo , Soja , Suínos/crescimento & desenvolvimento , Suínos/imunologia , Desmame , Zea mays
12.
JAMA Netw Open ; 3(4): e202895, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32297948

RESUMO

Importance: Immunotherapy using immune checkpoint inhibitors has been remarkably effective for treating multiple cancer types, and the gut microbiome is a possible factor affecting immune checkpoint inhibitor efficacy. However, the association between the gut microbiome and immune status of the tumor microenvironment remains unclear. Short-chain fatty acids (SCFAs) are major end product metabolites produced by the gut microbiota and have wide-ranging impacts on host physiology. Objective: To evaluate fecal and plasma SCFAs in patients with solid cancer tumors treated with programmed cell death-1 inhibitors (PD-1i). Design, Setting, and Participants: This was a prospective cohort biomarker study of patients with cancer who planned therapy with PD-1i at Kyoto University Hospital between July 2016 and February 2019. Data were analyzed from October 2019 to February 2020. Exposures: Patients who were treated with nivolumab or pembrolizumab were classified into 2 groups based on their treatment response using Response Evaluation Criteria in Solid Tumors version 1.1: responders who achieved an objective response and nonresponders. Dietary information in terms of intake frequency was obtained. Concentrations of SCFAs in fecal and plasma samples collected before PD-1i administration were measured using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry. Main Outcomes and Measures: The concentration of SCFAs and progression-free survival. Results: Among 52 patients enrolled, the median (range) patient age was 67 (27-84) years, and 23 (44%) were women. Median (range) duration of follow-up of the survivors after administration of PD-1i was 2.0 (0.4-4.1) years. The overall response rate was 28.8%. High concentrations of some SCFAs were associated with longer progression-free survival. These included fecal acetic acid (hazard ratio [HR], 0.29; 95% CI, 0.15-0.54), propionic acid (HR, 0.08; 95% CI, 0.03-0.20), butyric acid (HR, 0.31; 95% CI, 0.16-0.60), valeric acid (HR, 0.53; 95% CI, 0.29-0.98), and plasma isovaleric acid (HR, 0.38; 95% CI, 0.14-0.99). Conclusions and Relevance: Results of this study suggest that fecal SCFA concentrations may associated with PD-1i efficacy; thus, SCFAs may be the link between the gut microbiota and PD-1i efficacy. Because fecal examinations are completely noninvasive, they may be applicable for routine monitoring of patients.


Assuntos
Anticorpos Monoclonais Humanizados/efeitos adversos , Anticorpos Monoclonais Humanizados/uso terapêutico , Antineoplásicos Imunológicos/efeitos adversos , Microbioma Gastrointestinal/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Nivolumabe/efeitos adversos , Nivolumabe/uso terapêutico , Adulto , Idoso , Idoso de 80 Anos ou mais , Antineoplásicos Imunológicos/uso terapêutico , Ácidos Graxos Voláteis/análise , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos
13.
J Anim Sci ; 98(5)2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32271920

RESUMO

The objectives of this study were to compare ruminal total tract digestibility, bacterial communities, and eating and rumination activity between Holstein and Angus steers fed grain- or forage-based diets. Six Holstein steers (average body weight [BW] = 483 ± 23 kg) and six Angus steers (average BW = 507 ± 29 kg), previously fitted with rumen cannulae, were fed in a crossover design with a 2 × 2 factorial arrangement of four treatments: 1) Holsteins fed a grain-based diet, 2) Holsteins fed a forage-based diet, 3) Angus fed a grain-based diet, and 4) Angus fed a forage-based diet. Each period was 35 d with 26 d of diet adaptation and 9 d of sample collection. On days 1 and 2 of collection, feeding activity was recorded for 48 h. On day 3, rumen contents were sampled to measure ruminal pH at 0, 3, 6, 12, and 18 h after feeding. A portion of the strained ruminal fluid was subsampled at 0, 3, and 6 h for volatile fatty acids (VFA) analysis. Rumen contents were subsampled at 3 h for analysis of bacterial communities. From day 4 to 8, total fecal excretion, feed, and refusals samples were collected and analyzed for dry matter (DM), neutral detergent fiber (NDF), and starch. On days 8 and 9 (0 and 3 h post-feeding, respectively), total reticulorumen evacuation was conducted and contents were weighed. Data were analyzed using the MIXED procedures in SAS (v9.4 SAS Inst. Inc., Cary, NC). Repeated measures were used to analyze changes in ruminal pH and VFA over time. There were no interactions of diet × breed (P ≥ 0.07). While the main effects of diet were expected, unique to these data is the fact that bacterial diversity and richness were reduced (P < 0.01) in cattle fed grain-based diets. There was no main effect (P > 0.34) of breed on total tract DM, organic matter, and starch digestibility, but Angus cattle had greater (P = 0.01) NDF digestibility than Holsteins. The increased NDF digestibility may be associated with a numerical (P = 0.08) increased numbers of bacterial species in Angus steers compared with Holstein steers. Holstein steers also spent more time (P ≤ 0.05) ruminating than Angus steers. There was no effect (P > 0.80) of breed on reticulorumen content at feeding time; however, Holstein steers had greater (P = 0.04) reticulorumen content on a wet basis 3 h post-feeding. Although Holstein steers spent more time ruminating, Angus steers were better able to digest NDF when compared with Holsteins, regardless of basal diet, and this improvement may be related to changes in bacterial communities in the rumen or to rumination activity.


Assuntos
Ração Animal/análise , Bovinos/fisiologia , Fibras na Dieta/metabolismo , Ácidos Graxos Voláteis/análise , Microbioma Gastrointestinal , Animais , Peso Corporal , Cruzamento , Bovinos/microbiologia , Estudos Cross-Over , Dieta/veterinária , Digestão , Ingestão de Alimentos , Grão Comestível , Fermentação , Trato Gastrointestinal/microbiologia , Trato Gastrointestinal/fisiologia , Masculino , Rúmen/microbiologia , Rúmen/fisiologia , Amido/metabolismo
14.
PLoS One ; 15(4): e0231533, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32282837

RESUMO

Most of the research efforts involving the bovine gastrointestinal microbiota have focused on cattle's forestomach, particularly the rumen, so information concerning the bovine fecal microbiota is more scarce, especially in young beef cattle. The present study was performed to evaluate the ruminal and fecal microbiotas of beef calves as they reached the end of their nursing phase. A total of 18 Angus cow/calf pairs were selected and assigned to one of two treatment groups for the last 92 days of the calves' nursing period, as follows: 1) calves were supplemented with concentrate in a creep feeding system; or 2) control group with no supplementation of calves. After 92 days, ruminal and fecal samples were individually obtained from calves in both groups, and their microbiotas were evaluated using 16S rRNA gene sequencing. Ruminal samples were predominated by Prevotella (18 to 23% of the total bacterial abundance), regardless if calves received supplementation or not; however, in the feces, Prevotella was only the seventh most abundant genus (0.6 to 2.1% of total bacterial abundance). Both the rumen (P = 0.01) and the feces (P = 0.05) of calves that received supplementation had greater abundance of Firmicutes. In addition, calves that were supplemented had lower abundance of Fibrobacteres (P = 0.03) in their rumens. Regardless if the calves were supplemented or not, Faith's Phylogenetic Diversity index (P ≤ 0.007) and total concentration of short chain fatty acids (P < 0.001) were both greater in the rumen than in the feces of calves. In summary, the ruminal and fecal microbiotas of weanling beef calves were considerably distinct. Additionally, supplementation with creep feed caused some significant changes in the composition of the gastrointestinal microbiota of the calves, especially in the rumen, where supplementation caused an increase in Firmicutes and a decrease in abundance of Fibrobacteres.


Assuntos
Ração Animal , Bovinos/microbiologia , Fezes/microbiologia , Rúmen/microbiologia , Fenômenos Fisiológicos da Nutrição Animal , Animais , Biodiversidade , Ácidos Graxos Voláteis/análise , Feminino , Masculino , Desmame , Ganho de Peso
15.
Artigo em Inglês | MEDLINE | ID: mdl-32193004

RESUMO

Short-chain fatty acids (SCFAs) are gut microbiota metabolites recognized for their beneficial effects on the host organism. In this study, a simple and rapid sample preparation method combined to SCFAs analysis by direct injection and gas chromatography coupled with flame ionization detection (GC-FID), for the determination and quantification of eight SCFAs (acetic, propionic, i-butyric, butyric, i-valeric, valeric, i-caproic and caproic acids) in rat, mice and human faeces and in fermentation fluids samples, has been developed and validated. The method consists of extraction of the SCFAs by ethyl ether after acidification of the samples. The effect of the number of extractions has been assessed in order to optimize the procedure and to obtain a satisfactory yield for all the analyzed SCFAs. The increase of the extracted analytes quantity was significant passing from 1 to 2 and from 2 to 3 extractions (P < 0.05), while no significant differences were found performing 3, 4 or 5 extractions (P > 0.05). The SCFAs extracted are directly analyzed by GC-FID without derivatization and separated on a polyethylene glycol nitroterephthalic acid modified coated capillary column, with a chromatographic run time of 13 min. The proposed method showed good sensitivity, with limits of quantifications in the range 0.14-0.48 µM for SCFAs from propionic to caproic acids and 2.12 µM for acetic acid; recovery was between 80.8 and 108.8% and intraday and interday repeatability in the range 0.6-5.0% of precision (RSD, %) The optimized method is suitable for the quantitative analysis of SCFAs in real samples of rat, mouse and human faeces and in fermentation fluids, and it can be applied also to very small amount of faecal sample (20 mg).


Assuntos
Misturas Complexas/análise , Ácidos Graxos Voláteis/análise , Ácidos Graxos Voláteis/metabolismo , Fezes/química , Animais , Cromatografia Gasosa , Misturas Complexas/metabolismo , Éter/química , Fermentação , Microbioma Gastrointestinal , Humanos , Limite de Detecção , Metabolômica/métodos , Camundongos , Polietilenoglicóis/química , Ratos , Reprodutibilidade dos Testes
16.
Carbohydr Polym ; 235: 115904, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32122473

RESUMO

In vitro digestive conditions were simulated to investigate the digestibility of polysaccharides prepared from squash (SPS). A small amount of free glucose monosaccharide was released after salivary and intestinal digestion due to the breakdown of α-(1 → 4)-glucose linkages and may form SPS or a starch impurity. At the same time, there was no obvious change in molecular weight distribution and reducing sugar content throughout this digestion period, demonstrating that the main structure of SPS was relatively stable under the simulated digestive conditions. Thus, most SPS can be transported intact to the large intestine. In addition, SPS alleviated type 2 diabetes (T2D) in rats. Moreover, the content of short-chain fatty acids (SCFAs) in the colon significantly increased after treatment with SPS. The present research provides insight into the non-digestibility of SPS, and suggests its utility to alleviate T2D by promoting the production of SCFA in the colon.


Assuntos
Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Ácidos Graxos Voláteis/biossíntese , Polissacarídeos/metabolismo , Animais , Colo/química , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Tipo 2/induzido quimicamente , Ácidos Graxos Voláteis/análise , Intestinos/química , Masculino , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Ratos , Ratos Wistar , Saliva/química , Saliva/metabolismo , Estômago/química , Estreptozocina/administração & dosagem
17.
Benef Microbes ; 11(2): 191-200, 2020 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-32208927

RESUMO

The aim of the study was to investigate the prebiotic potential of xylo-oligosaccharides (XOS) from sugarcane in a validated, dynamic, computer-controlled in vitro model of the colon (TIM-2) simulating human adults. In two sets of experiments, each with a different microbiota, 3 different doses of XOS were tested at 1.0 g/day, 1.5 g/day and 3.0 g/day. The in vitro model was run for 72 h, and at the start and subsequently every 24 h samples were taken and analysed for short-chain fatty acids (SCFA) and gut microbiota composition. SCFA were analysed using ion chromatography, whereas microbiota composition was analysed using sequencing of the V3-V4 region of the 16S rRNA gene. XOS showed a similar SCFA production per gram of substrate as the control medium, including butyrate, which is considered to be important for gut health. In both sets of experiments XOS showed a consistent dose-dependent increase in abundance over time of the genus Bifidobacterium, and within that of the species B. adolescentis and an unidentified species (labelled 'sp.1'). The results show the potential prebiotic effect of XOS from sugarcane, by its capacity to generate butyrate and increase the health-beneficial bifidobacteria.


Assuntos
Colo/microbiologia , Microbioma Gastrointestinal , Modelos Biológicos , Oligossacarídeos/química , Prebióticos/análise , Saccharum/química , Adulto , Butiratos/análise , Ácidos Graxos Voláteis/análise , Humanos , Técnicas In Vitro
18.
J Dairy Sci ; 103(5): 4702-4716, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32171513

RESUMO

We investigated changes in rumen fermentation, the fluid bacterial community, and predicted functional pathway profiles in Holstein cows with and those without subacute ruminal acidosis (SARA) during the periparturient period. Eighteen multiparous Holstein cows categorized in the SARA (n = 9) or non-SARA (n = 9) groups depending on whether they developed SARA during the 2 wk after parturition. Reticulo-ruminal pH was measured continuously throughout the study. Rumen fluid and blood samples were collected 3 wk before and 2 and 6 wk after parturition, with an additional blood sample collected during the 0 and 4 wk after parturition. The 7-d mean reticulo-ruminal pH was significantly decreased in both groups postpartum compared with prepartum, with a greater and longer lasting change in the SARA group. The postpartum total volatile fatty acid concentration was significantly higher in the non-SARA than in SARA cows. Rumen bacterial richness and diversity were not affected by the periparturient period, but the non-SARA group had a significantly higher Simpson diversity index postpartum. No significant change was observed in the composition of major bacterial phyla or genera during the periparturient period. Among 17 operational taxonomic units (OTU) shared by all cows, the relative abundances of OTU36 (genus Pseudobutyrivibrio) in the SARA group and OTU4 (genus Prevotella) in the non-SARA group decreased significantly during the postpartum period compared with prepartum. In contrast, non-SARA cows had more diverse core bacterial OTU community including 23 additional OTU than those with SARA including 3 additional OTU (40 vs. 20 OTU). The OTU36 was significantly correlated with the proportion of iso-valeric acid (r = -0.353), and OTU4 was significantly correlated with the 7-d mean reticulo-ruminal pH (r = 0.432) and the proportions of acetic acid (r = 0.353) and propionic acid (r = -0.380). Prepartum, OTU2 (genus Prevotella) was the only OTU that differed significantly between the 2 groups. Changes in predicted functional pathway were primarily identified during the postpartum period, with 2 pathways upregulated at 3 wk prepartum and 24 upregulated and 34 downregulated at 6 wk postpartum in the SARA group. Our results suggest that SARA incidence is not strictly related to changes in rumen fermentation or fluid bacterial community structure, but the predicted functional pathways of bacterial communities showed late responses to a postpartum decrease in reticulo-ruminal pH. Therefore, postpartum predicted functional pathway analysis may indicate the underlying mechanisms causing SARA in Holstein cows during the periparturient period.


Assuntos
Acidose/veterinária , Bactérias/isolamento & purificação , Doenças dos Bovinos/microbiologia , Ácidos Graxos Voláteis/análise , Microbioma Gastrointestinal , Acidose/metabolismo , Acidose/microbiologia , Animais , Bactérias/genética , Bovinos , Doenças dos Bovinos/metabolismo , Dieta/veterinária , Feminino , Fermentação , Concentração de Íons de Hidrogênio , Parto , Gravidez , Prevotella/genética , Prevotella/isolamento & purificação , Rúmen/metabolismo , Rúmen/microbiologia
19.
J Anim Sci ; 98(3)2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32076715

RESUMO

The in vitro gas production technique (IVGPT) has been a valuable tool in ruminant nutrition research for decades and has more recently been used in horse nutrition studies to investigate fermentation activities of the equine hindgut though primarily using feces as inoculum. This study was conducted to evaluate the use of equine rectal content in the IVGPT system as a viable inoculum that can be considered representative of the activities throughout the equine hindgut. Additionally, the study was conducted to measure the effects on fermentation kinetics and end-product production using inoculum from horses fed supplemental levels of coated sodium butyrate in an IVGPT system. Eight warmblood horses were fed a diet consisting of haylage (1% DM intake based on ideal body weight [BW]) and a mash concentrate formulated to provide 2.5 g nonstructural carbohydrate (NSC)/kg BW per meal. The diet was intended to create a NSC challenge to the microbial populations of the hindgut. The horses were randomly assigned to treatment or control group and after a 1-wk diet-adaptation period, the treatment group received 0.4 g/kg BW per day of a coated sodium butyrate supplement, while the control group received a placebo (coating only). After a 3-wk treatment period, the animals were sacrificed and digesta from the cecum, left ventral colon, right dorsal colon, and the rectum were collected within 30 min postmortem and used as inocula for the IVGPT trial. Haylage and concentrates fed to the test animals were also used as substrates in vitro. Sodium butyrate supplementation was not significant for gas production parameters or VFA measured suggesting no effect of sodium butyrate supplementation on the extent or kinetics of gas production or microbial end-product production (P ≥ 0.073). Differences in inocula were significant for organic matter corrected cumulative gas production (P = 0.0001), asymptotic gas production of the second phase (A2) (P < 0.0001); and maximal rate of OM degradation of the second phase (Rmax2) (P = 0.002). Inocula had a significant effect on total VFA (P = 0.0002), butyrate (Bu) (P = 0.015), branched chain fatty acids (P < 0.0001), pH (P < 0.0001), and ammonia (NH3) (P = 0.0024). In conclusion, based on observed results from this study, total tract digestibility may be overestimated if using rectal content inoculum to evaluate forage-based feeds in an IVGPT system.


Assuntos
Ácidos Graxos Voláteis/metabolismo , Microbioma Gastrointestinal/fisiologia , Cavalos/microbiologia , Animais , Ácido Butírico/análise , Dieta/veterinária , Suplementos Nutricionais/análise , Digestão , Ácidos Graxos Voláteis/análise , Fezes/microbiologia , Fermentação , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/microbiologia , Cavalos/metabolismo , Técnicas In Vitro/veterinária , Distribuição Aleatória , Reto/metabolismo , Reto/microbiologia
20.
PLoS One ; 15(2): e0229281, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32092087

RESUMO

The aim of this study was to investigate the effect on growth performance and nutrient utilisation when supplementing diets deficient in energy and protein with carbohydrase enzymes or xylo-oligosaccharide in broilers challenged with coccidia. 960 Ross 308 broilers were used in this 21-day study. The treatments were arranged into a 2×4 factorial with 2 challenge states (challenged and non-challenged) and 4 different additive types (control, xylanase alone, xylanase and ß-glucanase mixture and xylo-oligosaccharide). On day 14, the challenged group received 12× the recommended dose of coccidiosis vaccine while the non-challenged group received a sham treatment of water only. The birds and feed were weighed on days 0, 14 and 21. On day 21, two birds per pen were euthanized, the caeca were removed and the contents collected for short chain fatty acid analysis. Six more birds per pen were euthanized and ileal digesta were collected and pooled per pen for nutrient digestibility analysis. Feed intake was greater (P < 0.05) on days 14 and 21 when xylo-oligosaccharide was included in the diet compared to the xylanase and ß-glucanase mixture in birds challenged with coccidiosis. Including xylo-oligosaccharide in the diet improved (P < 0.05) the digestibility of nitrogen and supplementing diets with the xylanase and ß-glucanase mixture improved (P < 0.05) the digestibility of several amino acids. The concentration of arabinose and xylose was (P < 0.001) greater when broiler diets were supplemented with carbohydrase enzymes or xylo-oligosaccharide compared to the control. Although there was an increase in short chain fatty acid production due to the addition of carbohydrase enzymes or xylo-oligosaccharide, there was no additive effect on the %G+C profile of caecal bacteria however there was a negative effect of coccidiosis. In conclusion, the similarity in the response to carbohydrase enzymes or xylo-oligosaccharide supplementation illustrates that the hydrolysis products from carbohydrase activity may have prebiotic like effects.


Assuntos
Galinhas/crescimento & desenvolvimento , Coccidiose , Glicosídeo Hidrolases/farmacologia , Prebióticos , Fenômenos Fisiológicos da Nutrição Animal , Animais , Galinhas/parasitologia , Coccidiose/dietoterapia , Coccidiose/tratamento farmacológico , Dieta , Suplementos Nutricionais , Digestão , Grão Comestível , Ácidos Graxos Voláteis/análise , Glicosídeo Hidrolases/metabolismo , Oligossacarídeos/farmacologia
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