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BACKGROUND AND AIMS: Non-alcoholic steatohepatitis (NASH) has multifactorial etiopathogenesis, and intestinal microbiota is co-responsible in this process. The aim of this study was to evaluate the intestinal microbiota in NASH patients with different metabolic profiles. METHODS: Patients with biopsy-proven NASH were evaluated. Subjects were divided into two groups according to their metabolic profile, with or without metabolic syndrome (MS). Their characteristics in relation to liver disease and intestinal microbiota were analyzed. To evaluate the microbiota, breath tests to investigate small intestinal bacterial overgrowth (SIBO) and fecal microbiota analysis by fluorescence in situ hybridization (FISH) were performed. RESULTS: There was a high prevalence of SIBO in both groups, with no significant difference between them. Breathing tests were positive in 43.8% of patients with MS and 50% of those without MS. There was a significant difference regarding the quantification of Verrucomicrobiales, less abundant in patients with NASH without MS. Its lower concentration also correlated with higher serum ferritin levels and higher hepatocyte ballooning. This order of bacteria, through its representative in human microbiota, Akkermansia muciniphila, is associated with mucosal protection and metabolic processes with liver aggression. CONCLUSIONS: Our results suggested that lower Verrucomicrobiales concentration is associated with higher inflammatory activity in patients with NASH without MS, where the disease etiopathogenesis does not have its classic metabolic substrate.

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Information about bacterial diversity, such as the number of each species in the root canals of primary teeth, contributes to improving our effective management of infections of endodontic origin in primary teeth. This study made a qualitative and quantitative assessment of the bacteria in the root canals of primary teeth with necrotic pulp, using the fluorescence in situ hybridization (FISH) technique. Thirty-one primary teeth with pulp necrosis from 31 children were evaluated using the FISH technique, to detect the presence and density of Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Enterococcus faecalis, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Streptococcus, Streptococcus mutans, Streptococcus sobrinus, Tannerella forsythia and Treponema denticola. Descriptive measures explained the data related to density, and Student's t-test assessed the differences among the densities of each bacterium, according to signs and symptoms. The bacterial density was paired and correlated. All bacteria tested were detected and identified in all the samples. The average number of bacterial individuals from each species ranged from 1.9 x 108 cells/mL (S. mutans) to 3.1 x 108 cells/mL (F. nucleatum) (p > 0.05). The sum of the mean counts of each bacterium represented almost 80% of the entire microbial community. Patients with pain had significantly more T. denticola, and those with edema showed a greater density of Streptococcus and P. nigrescens (p < 0.05). This study revealed that all 12 bacteria evaluated were found in all primary teeth with pulp necrosis. There was no predominance among the species studied; all species had a similar number of individuals.

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OBJECTIVE: The aim of this study was to verify the effect of a beverage containing ora-pro-nobis (OPN) flour on intestinal microbiota, gastrointestinal symptoms, and anthropometric parameters in women. METHODS: This prospective, double-blinded, randomized clinical trial included 24 women volunteers. For 6 wk, the test group received a beverage supplemented with OPN, and the control group received the same beverage without OPN. Fecal microbiota were evaluated by the fluorescence in situ hybridization method. Each participant responded to a questionnaire based on the Gastrointestinal Symptom Rating Scale. The data were analyzed by Student's t test to compare the effects between treatments (P < 0.05). RESULTS: Supplementation with the OPN beverage reduced weight (P = 0.008), waist circumference (P = 0.010), and body fat (P = 0.000). Additionally, the treatment increased satiety (P = 0.039), reduced eructation (P = 0.038) and constipation (P = 0.017), and improved feces consistency (P = 0.017). CONCLUSION: The OPN flour beverage has an effect on weight reduction, body fat composition, and improvement in gastrointestinal symptoms. These results highlight the beneficial effects of ora-pro-nobis flour beverage against the development of obesity.

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We studied petroleum biodegradation by biostimulation by using water in oil in water (W/O/W) double emulsions. These emulsions were developed using seawater, canola oil, surfactants, and mineral salts as sources of NPK. The emulsions were used in the simulation of hydrocarbon bioremediation in oligotrophic sea water. Hydrocarbon biodegradation was evaluated by CO2 emissions from microcosms. We also evaluated the release of inorganic nutrients and the stability of the emulsion's droplets. The double emulsions improved CO2 emission from the microcosms, suggesting the increase in the hydrocarbon biodegradation. Mineral nutrients were gradually released from the emulsions supporting the hydrocarbon biodegradation. This was attributed to the formation of different diameters of droplets and therefore, varying stabilities of the droplets. Addition of the selected hydrocarbonoclastic isolates simulating bioaugmentation improved the hydrocarbon biodegradation. We conclude that the nutrient-rich W/O/W emulsion developed in this study is an effective biostimulation agent for bioremediation in oligotrophic aquatic environments.

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Abstract Information about bacterial diversity, such as the number of each species in the root canals of primary teeth, contributes to improving our effective management of infections of endodontic origin in primary teeth. This study made a qualitative and quantitative assessment of the bacteria in the root canals of primary teeth with necrotic pulp, using the fluorescence in situ hybridization (FISH) technique. Thirty-one primary teeth with pulp necrosis from 31 children were evaluated using the FISH technique, to detect the presence and density of Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Enterococcus faecalis, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Streptococcus, Streptococcus mutans, Streptococcus sobrinus, Tannerella forsythia and Treponema denticola. Descriptive measures explained the data related to density, and Student's t-test assessed the differences among the densities of each bacterium, according to signs and symptoms. The bacterial density was paired and correlated. All bacteria tested were detected and identified in all the samples. The average number of bacterial individuals from each species ranged from 1.9 x 108 cells/mL (S. mutans) to 3.1 x 108 cells/mL (F. nucleatum) (p > 0.05). The sum of the mean counts of each bacterium represented almost 80% of the entire microbial community. Patients with pain had significantly more T. denticola, and those with edema showed a greater density of Streptococcus and P. nigrescens (p < 0.05). This study revealed that all 12 bacteria evaluated were found in all primary teeth with pulp necrosis. There was no predominance among the species studied; all species had a similar number of individuals.

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BACKGROUND AND AIMS: Modulation of the gut microbiota emerges as a therapeutic possibility to improve health. Our objective was to compare the impact of three months of intervention with diet plus nutritional orientation versus only nutritional orientation on the gut microbiota and metabolic-nutritional profile of outpatients with non-alcoholic steatohepatitis. METHODS: It was a randomized clinical trial with 40 outpatients (49.48 ± 10.3 years), allocated in two groups: DIET group (n=20), who received diet (1.651.34 ± 263.25 kcal; 47% carbohydrates, 28% lipids, 25% proteins, 30 g fibers) and nutritional orientation, and control group (n = 20), which received only nutritional orientation. RESULTS: The DIET group, in relation to baseline, presented a reduction in body weight (p<0.001), BMI (p<0.001), waist circumference (p=0.001), percentage of fat (p=0.002), serum aspartate aminotransferase (p<0.001), alanine aminotransferase (p<0.001), γ-glutamyltransferase (p=0.001), glycemia (p=0.003), homeostasis model assessment of insulin resistance (p=0.017), total cholesterol (p=0.014), and triacylglycerols (p=0.008), whereas the control group did not present changes. After intervention, the small intestinal bacterial overgrowth frequency was 30% in the DIET group and 45% in the control group (p=0.327). In the DIET group, an increase in the density of total microorganisms (3.76 ± 7.17 x 10 8 cells g -1 ; p=0.048) was detected, while in the control group reduced Bacteroidetes (-0.77 ± 2.01 x 10 8 cells g -1 , p=0.044) and Verrucomicrobiales (-0.46 ± 0.75 x 10 8 cells g -1 ; p=0.022) were observed. CONCLUSIONS: The results suggest that exclusively dietary modifications contribute to health promotion in non-alcoholic steatohepatitis and should be the basis of nutritional treatment for this condition.

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Although lifestyle and physiology in obese individuals are accepted to lead to changes in the intestinal microbiota, uncertainty remains about microbiota dysbiosis, and xenobiotics intake, as a source of selective pressure, independent of antimicrobial chemotherapy. The aim of this study was to compare the occurrence of antimicrobial resistance genetic markers (ARG) in faecal specimens of eutrophic, overweight and obese individuals, and their correlation with xenobiotic intake and gut bacteria density. Methods: This was a cross-sectional case-controlled study including 72 adult participants with no record of intestinal or systemic diseases, or recent use of antimicrobials, grouped as eutrophic, overweight, or obese. Anthropometric profile, eating habits and oral xenobiotics intake were recorded. Faecal metagenomic DNA was used to screen for ARG by PCR, and to measure bacterial groups by fluorescence in situ hybridization (FISH). Student's t and Wilcoxon tests were used to compare means and differences in ARG detection (95% confidence intervals). Correlation analyses (odds ratio) and relationships between bacteria density and ARG were determined. Results: Increase in abdominal circumference, waist circumference, hip, waist-hip ratio, BMI, carbohydrate, fibres, and total calorie intakes were different from eutrophic to obese participants. Habitual use of antihypertensive and anti-inflammatory drugs, antacids, and artificial sweeteners were associated mainly with obesity and overweight. Nutritional supplements were associated to the eutrophic group. ARG screening showed differences being more frequent among obese, and positive for 27 genetic markers related to ß-lactams, tetracyclines, the macrolide lincosamide and streptogramin group, quinolones, sulfonamides, aminoglycosides, and efflux pump. Positive correlation between ARG and BMI, caloric intake, and intake of xenobiotics, was observed for obese individuals. Relationships among ARG detection and bacteria densities were also different. Conclusions: This study reinforces the hypothesis that obese individuals may harbour an altered gut microbiota, if compared to eutrophic. The overweight individuals display a transitional gut microbiota which seems to be between eutrophic and obese. Furthermore, the increased xenobiotic intake associated to obesity may play an important role in the antimicrobial resistance phenomenon.

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Probiotics are widely used in the food industry and may affect the oral microbiota. This study aimed to evaluate the effect of petit-suisse plus probiotic on the microbiota of children's saliva. Strawberry flavor petit-suisse cheese plus green banana flour without Lactobacillus casei (control) and with addition of Lactobacillus casei (probiotic) were prepared and used in the experiments. The feasibility of adding probiotic to petit-suisse was assessed over 28 days. Saliva samples from volunteers who consumed the petit-suisses produced were collected before the period of consumption of the products (T0), on the last day of consumption of petit-suisse (T1) and two weeks after the consumption stopped (T2 - post treatment). The samples were immediately fixed with paraformaldehyde in a final concentration of 2%, after which the bacteria of interest were identified by Fluorescent In Situ Hybridization (FISH) and counted with the aid of an epifluorescence microscope. Lactic bacteria were viable between 108 and 109 CFU·g-1 for 28 days of storage at 4 °C. Both products (control and with L. casei added) were able to significantly reduce (p < 0.05) the total number of microorganisms and Streptococcus mutans in the volunteers' saliva. However, only the product plus L. casei decreased density of Agreggatibacter actinomycetemcomitans, as well as being able to maintain the low density of Porphyromonas gingivalis post treatment. Therefore, the petit-suisse cheese developed showed the ability to carry probiotic microorganism, being a potential alternative for reducing potentially pathogenic microbiota in the oral cavity.

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Litopenaeus vannamei is the most cultured marine shrimp in all types of systems including the Bioflocs Technology System (BFT). Bioflocs are formed by microorganisms, among these, autotrophic bacteria are responsible for the nitrification process. This study aimed to identify and promote the development of nitrifying bacteria by adding artificial substrates and biofloc inoculum in L. vannamei culture in a BFT system. The experiment consisted of four treatments with three replics (4x3) as follows: (1) Control: clear water in which bioflocs were formed; (2) IN (10%): clear water with biofloc inoculum (10%); (3) IB: clear water with substrate (immature bioballs); and (4) MB: clear water with mature bioballs inoculum from a recirculation system. Treatments were stocked with shrimp juveniles (4.92±0.45 g) in 12 tanks with 200 L working volume at a stocking density of 200 shrimp/m³. Shrimps were fed twice a day with a commercial feed (38% CP) following a feeding table, and daily observations intake were made over the four weeks of the experiment. Biofloc and bioballs samples were collected to detect the growth of the population of nitrifying and heterotrophic bacteria by FISH. There was no significant difference between treatments (P>0.05) for survival, obtaining mean values greater than 88%. The IN (10%) treatment had lower concentrations of ammonia and nitrite, and nitrate concentration increased, while MB had a higher weight and biomass final, productivity, weekly weight gain and lower conversion of apparent feed for production performance results.(AU)

Litopenaeus vannamei é o camarão marinho mais cultivado em todos os tipos de sistemas, incluindo o Sistema de Tecnologia de Bioflocos (BFT). Os bioflocos são formados por microorganismos, entre estes, bactérias autotróficas que são responsáveis pelo processo de nitrificação. Este estudo teve como objetivo identificar e promover o desenvolvimento de bactérias nitrificantes pela adição de substratos artificiais e inóculo de bioflocos no cultivo de L. vannamei em sistema BFT. O experimento consistiu de quatro tratamentos com três repetições (4x3), sendo: (1) Controle: água limpa, na qual foram formados os bioflocos; (2) IN (10%): água limpa com inóculo de bioflocos (10%); (3) BI: água limpa com substrato (bioballs imaturos); e (4) BM: água limpa com inóculo de bioballs maduros de um sistema de recirculação. Para tanto, os tratamentos foram estocados com juvenis de camarão (4,92±0,45 g) em 12 tanques com 200 L de volume útil com densidade de 200 camarões/m³. Os camarões foram alimentados duas vezes ao dia com ração comercial (38% PB) seguindo uma tabela de alimentação, e observações diárias foram feitas ao longo das quatro semanas de experimento. Amostras de bioflocos e bioballs foram coletadas para detectar o crescimento da população de bactérias nitrificantes e heterotróficas por FISH. Não houve diferença significativa entre os tratamentos (P>0,05) para sobrevivência, obtendo-se valores médios superiores a 88%. O tratamento com IN (10%) apresentou menores concentrações de amônia e nitrito, e a concentração de nitrato aumentou, enquanto o BM apresentou maior peso e biomassa final, produtividade, ganho de peso semanal e menor conversão alimentar aparente para resultados de desempenho zootécnico.(AU)

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