Agave fructans enhance the effects of fermented milk products on obesity biomarkers: a randomised trial.

Dysbiosis has been implicated in childhood obesity. Oral intake of fermented milk containing Lacticaseibacillus casei strain Shirota preserves gut microbiota (GM) diversity in children and adults. This study was a double-blind trial involving 37 overweight or obese children aged 6-10 years. Children were followed over a 6-week intervention period in which they received different fermented milk products containing L. casei Shirota: 10 in the first group received just L. casei Shirota; 13 received L. casei Shirota with 3 g/day of inulin (L. casei+inulin); and 14 received L. casei Shirota with 3 g/day of fructans from Agave salmiana (L. casei+fructans). Principal component analysis showed the relationship between microbial abundance, GM metabolites, and other obesity-related markers. Supplementation with probiotics and synbiotics improved the HDL-cholesterol levels of overweight and obese children, although no changes in body composition were detected. We observed an increase in butyrate or propionate concentrations in the L. casei+fructans group compared to the end of the intervention (P<0.03). A diminished level of ANGPTL4 within the L. casei+fructans group (P=0.04) was also found, but no differences when lipopolysaccharide-binding protein was evaluated. The FFAR2+ cell frequency decreased between baseline and at the end of 6-week intervention in L. casei+inulin (P=0.02) and L. casei+fructans groups (P=0.04). In contrast, the percentage of CD14+FFAR3+ frequency increased in the same groups (P=0.04). The L. casei Shirota with inulin or fructans modulates GM, which improves the lipid profile and changes at a molecular level, such as expression of FFAR3 and FFAR2, ANGPTL4, propionate, and butyrate. It, therefore, could be considered an interesting therapeutic possibility for treating childhood overweight and obesity. The study was registered at ClinicalTrials.gov (ID: NCT05423015).

Hydrogenotrophic methanogens dominate in biogas reactors fed with defined substrates.

Methanogenic communities in 200L biogas reactors containing liquid manure were investigated for 33 d. The reactors were consecutively fed with casein, starch and cream. Real-time PCR with primers targeting the gene for methyl coenzyme-M reductase (mcrA) resulted in copy numbers of up to 2.1×10(9) g dry mass(-1). Single strand conformation polymorphism (SSCP) analysis revealed a stable community consisting of few hydrogenotrophic methanogens. One of the two most abundant species was closely related to Methanospirillum hungatei, whereas the other one was only distantly related to other methanogens, with Methanopyrus kandleri being the closest cultivated relative. Most probable number (MPN) cultivations were accomplished with a sample from a 600 m(3) reactor from which all manures used in the experiments originated, and equal cell counts of ca. 10(9) g dry mass(-1) were found for cultivations with acetate, H(2) and methanol. SSCP analysis of these samples and sequencing of the DNA bands identified different hydrogenotrophic methanogens in all samples, and acetoclastic methanogens closely related to Methanosarcina mazei in the samples cultivated with acetate and methanol. As the acetoclastic species were not found in any other SSCP sample, it was supposed that the ammonia values in the manure of the laboratory biogas reactor, which ranged from 2.48 to 3.61 g NH(4)-NL(-1), inhibited the growth of the acetoclastic methanogens.

Unexpected stability of Bacteroidetes and Firmicutes communities in laboratory biogas reactors fed with different defined substrates.

In the present study, bacterial communities in 200-liter biogas reactors containing liquid manure consecutively fed with casein, starch, and cream were investigated over a period of up to 33 days. A 16S rRNA gene clone library identified Bacteroidetes and Firmicutes as the most abundant bacterial groups in the starting material, at 58.9% and 30.1% of sequences, respectively. The community development of both groups was monitored by real-time PCR and single-strand conformation polymorphism (SSCP) analysis. The Firmicutes and Bacteroidetes communities were unexpectedly stable and hardly influenced by batch-feeding events. The continuous feeding of starch led to community shifts that nevertheless contributed to a stable reactor performance. A longer starving period and a change in the pH value resulted in further community shifts within the Bacteroidetes but did not influence the Firmicutes. Predominant DNA bands from SSCP gels were cloned and sequenced. Sequences related to Peptococcaceae, Cytophagales, and Petrimonas sulfuriphila were found in all samples from all experiments. Real-time PCR demonstrated the abundance of members of the phylum Bacteroidetes and also reflected changes in gene copy numbers in conjunction with a changing pH value and acetate accumulation.

Nitrate-dependent iron(II) oxidation in paddy soil.

Iron(III) profiles of flooded paddy soil incubated in the greenhouse indicated oxidation of iron(II) in the upper 6 mm soil layer. Measurement of oxygen with a Clark-type microelectrode showed that oxygen was only responsible for the oxidation of iron(II) in the upper 3 mm. In the soil beneath, nitrate could be used as electron acceptor instead of oxygen for the oxidation of the iron(II). Nitrate was still available 3 mm below the soil surface, and denitrifying activity was indicated by higher concentrations of nitrite between 3 and 6 mm soil depth. Nitrate was generated by nitrification from ammonium. Ammonium concentrations increased beneath 6 mm soil depth, indicating ammonium release and diffusion from deeper soil layers. High concentrations of ammonium were also found at the surface, probably resulting from N2 fixation by cyanobacteria. Experimental adjustment of the nitrate concentration in the flooding water to 200 microM stimulated nitrate-dependent iron(II) oxidation, which was indicated by significantly lower iron(II) concentrations in soil layers in which nitrate-dependent iron(II) oxidation was proposed. Soil incubated in the dark showed high iron(III) concentrations only in the layer where oxygen was still available. In this soil, the nitrogen pool was depleted because of the lack of N2 fixation by cyanobacteria. In contrast, soil incubated in the dark with 500 microM nitrate in the flooding water showed significantly higher iron(II) and significantly lower iron(II) concentrations in the anoxic soil layers, indicating nitrate-dependent iron(II) oxidation. Anoxic incubations of soil with nitrate in the flooding water also showed high concentrations of iron(II) and low concentrations of iron(II) in the upper 3 mm. As oxygen was excluded in anoxic incubations, the high iron(III) concentrations are a sign of the activity of nitrate-dependent iron(II) oxidizers. The presence of these bacteria in non-amended soil was also indicated by the most probable number (MPN) counts of nitrate-dependent iron(II) oxidizers in the layer of 3-4 mm soil depth, which revealed 1.6 x 10(6) bacteria g(-1) dry weight.