A newly isolated methanogen, strain DMS1T, is the first obligately anaerobic archaeon which was directly enriched and isolated from a freshwater sediment in defined minimal medium containing dimethyl sulfide (DMS) as the sole carbon and energy source. The use of a chemostat with a continuous DMS-containing gas stream as a method of enrichment, followed by cultivation in deep agar tubes, resulted in a pure culture. Since the only substrates utilized by strain DMS1T are methanol, methylamines, methanethiol (MT), and DMS, this organism is considered an obligately methylotrophic methanogen like most other DMS-degrading methanogens. Strain DMS1T differs from all other DMS-degrading methanogens, since it was isolated from a freshwater pond and requires NaCl concentrations (0 to 0.04 M) typical of the NaCl concentrations required by freshwater microorganisms for growth. DMS was degraded effectively only in a chemostat culture in the presence of low hydrogen sulfide and MT concentrations. Addition of MT or sulfide to the chemostat significantly decreased degradation of DMS. Transient accumulation of DMS in MT-amended cultures indicated that transfer of the first methyl group during DMS degradation is a reversible process. On the basis of its low level of homology with the most closely related methanogen, Methanococcoides burtonii (94.5%), its position on the phylogenetic tree, its morphology (which is different from that of members of the genera Methanolobus,Methanococcoides, and Methanohalophilus), and its salt tolerance and optimum (which are characteristic of freshwater bacteria), we propose that strain DMS1T is a representative of a novel genus. This isolate was named Methanomethylovorans hollandica. Analysis of DMS-amended sediment slurries with a fluorescence microscope revealed the presence of methanogens which were morphologically identical to M. hollandica, as described in this study. Considering its physiological properties, M. hollandica DMS1T is probably responsible for degradation of MT and DMS in freshwater sediments in situ. Due to the reversibility of the DMS conversion, methanogens like strain DMS1T can also be involved in the formation of DMS through methylation of MT. This phenomenon, which previously has been shown to occur in sediment slurries of freshwater origin, might affect the steady-state concentrations and, consequently, the total flux of DMS and MT in these systems.
Currently, studies reporting the digestibility of carbohydrates, starch and especially non‐starch polysaccharides (NSP) in fish are scarce. Carbohydrate digestibility in the diet is largely dependent upon carbohydrate composition (starch vs. NSP). NSP are often considered to be indigestible and thus of no nutritional value. The present study reviews carbohydrates in fish feed, distinguishing between total carbohydrate, starch and NSP. Besides a qualitative approach, a meta‐analysis was performed, compiling available data from digestibility studies on tilapia. Our meta‐analysis confirms the negative effect of NSP on performance (FCR) and nutrient digestibility (crude protein, fat and energy). However, an average NSP digestibility of 24.3% was calculated in 95 cases. Out of these 95 cases, 88% of them showed a positive NSP digestibility. NSP digestibility was shown to contribute to energy digestibility. The digestion of NSP in fish is associated with fermentation in the gut, producing beneficial volatile fatty acids that are rapidly absorbed by the colonic lumen. Therefore, in diet formulation, digestibility and thus energy originating from NSP should be taken into consideration because NSP contribute to the energy needs of fish, here tilapia. Besides being an energy source, specific types of NSP may have immune‐modulating and prebiotic effects and may be increasingly added to fish feed as modulators of fish health. We suggest that NSP is potentially (partly) digested by a wide range of fish species, especially by warm‐water species with a long gut adapted to feeding on plant matter, as these factors favour gut fermentation.
The experiment investigated whether the effect of enzyme supplementation on performance and nutrient digestibility is dependent on type of the ingredient(s) used, by incorporating wheat bran (WB), sunflower meal (SFM) and citrus pulp (CP) to a reference diet (REF). Those ingredients are known to be rich in different types of non‐starch polysaccharides (NSP). Diets were supplemented with and without (control) an enzyme mix (phytase 1,000 FTU/kg and xylanase U/kg) according to a 2 × 4 factorial arrangement. In total, 24 tanks (3 replicates/treatment) were used with 30 fish each (Nile tilapia, Oreochromis niloticus; mean initial body weight 41 g). Fish were restrictively fed the experimental diets for 43 days (80% of expected satiation). Enzyme supplementation affected the absolute growth (g/day) and FCR (p < 0.05), improving the growth of fish fed the WB and SFM diets, while fish fed the REF and CP diets did not benefit. NSP, energy, ash, phosphorous and calcium digestibility improved with enzyme supplementation (p < 0.05). There was an interaction effect on all growth parameters, as well as the digestibility of energy and phosphorus (p < 0.05). This indicates that the effectiveness of the enzymes was dependent on the NSP‐rich ingredient used and thus the composition of the NSP fraction.
From granular sludge of an upflow anaerobic sludge bed (UASB) reactor treating paper-mill wastewater, a sulfate-reducing bacterium (strain ASRB1) was isolated with acetate as sole carbon and energy source. The bacterium was rod-shaped, (1.4-1.9 x 2.5-3.4 microns), nonmotile, and gram-negative. Optimum growth with acetate occurred around 37 degrees C in freshwater medium (doubling time: 3.5-5.0 days). The bacterium grew on a range of organic acids, such as acetate, propionate, and butyrate, and on alcohols, and grew autotrophically with H2, CO2, and sulfate. Fastest growth occurred with formate, propionate, and ethanol (doubling time: approx. 1.5 days). Strain ASRB1 clusters with the delta subdivision of Proteobacteria and is closely related to Syntrophobacter wolinii, a syntrophic propionate oxidizer. Strain ASRB1 was characterized as a new genus and species: Desulforhabdus amnigenus.
Sustainable aquafeed production requires fishmeal replacement, leading to an increasing use of plant-derived ingredients. As a consequence, higher levels of antinutritional substances, such as non-starch polysaccharides and phytate, are present in aquafeeds, with negative effects on fish performance, nutrient digestibility and overall gut health. To alleviate these negative effects, providing exogenous digestive enzymes and/or probiotics can be an effective solution. In this study, we tested the effect of dietary supplementation of enzymes (phytase and xylanase) and probiotics (three strains of Bacillus amyloliquefaciens) on nutrient digestion kinetics and volatile fatty acid content along the gut, and the distal gut microbiome diversity in Nile tilapia. Chyme volatile fatty content was increased with probiotic supplementation in the proximal gut, while lactate content, measured for the first time in vivo in fish, decreased with enzymes along the gut. Enzyme supplementation enhanced crude protein, Ca and P digestibility in proximal and middle gut. Enzymes and probiotics supplementation enhanced microbial interactions as shown by network analysis, while increased the abundance of lactic acid bacteria and Bacillus species. Such results suggest that supplementation with exogenous enzymes and probiotics increases nutrient availability, while at the same time benefits gut health and contributes to a more stable microbiome environment.
From granular sludge of an upflow anaerobic sludge bed (UASB) reactor treating paper-mill wastewater, a sulfate-reducing bacterium (strain ASRB1) was isolated with acetate as sole carbon and energy source. The bacterium was rod-shaped, (1.4-1.9 x 2.5-3.4 microns), nonmotile, and gram-negative. Optimum growth with acetate occurred around 37 degrees C in freshwater medium (doubling time: 3.5-5.0 days). The bacterium grew on a range of organic acids, such as acetate, propionate, and butyrate, and on alcohols, and grew autotrophically with H2, CO2, and sulfate. Fastest growth occurred with formate, propionate, and ethanol (doubling time: approx. 1.5 days). Strain ASRB1 clusters with the delta subdivision of Proteobacteria and is closely related to Syntrophobacter wolinii, a syntrophic propionate oxidizer. Strain ASRB1 was characterized as a new genus and species: Desulforhabdus amnigenus.
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