Dynamics of mitochondrial adaptation and energy metabolism in rainbow trout (Oncorhynchus mykiss) in response to sustainable diet and temperature.

Impacts of plant-based ingredients and temperatures on energy metabolism in rainbow trout was investigated. A total of 288 fish (mean body weight: 45.6 g) were fed four isocaloric, isolipidic, and isonitrogenous diets containing 40% protein and 20% lipid and formulated as 100% animal-based protein (AP) and a blend of 50% fish oil (FO) and 50% camelina oil (CO); 100% AP and100% CO; 100% plant-based protein (PP) and a blend of 50% FO and 50% CO or 100% PP and 100% CO at 14 or 18 °C for 150 d. Diet did not significantly affect weight gain (WG) (P = 0.1902), condition factor (CF) (P = 0.0833) or specific growth rate (SGR) (P = 0.1511), but diet significantly impacted both feed efficiency (FE) (P = 0.0076) and feed intake (FI) (P = 0.0076). Temperature did not significantly affect WG (P = 0.1231), FE (P = 0.0634), FI (P = 0.0879), CF (P = 0.8277), or SGR (P = 0.1232). The diet × temperature interaction did not significantly affect WG (P = 0.7203), FE (P = 0.4799), FI (P = 0.2783), CF (P = 0.5071), or SGR (P = 0.7429). Furthermore, temperature did not influence protein efficiency ratio (P = 0.0633), lipid efficiency ratio (P = 0.0630), protein productive value (P = 0.0756), energy productive value (P = 0.1048), and lipid productive value (P = 0.1386); however, diet had significant main effects on PER (P = 0.0076), LPV (P = 0.0075), and PPV (P = 0.0138). Temperature regimens induced increased activities of mitochondrial complexes I (P = 0.0120), II (P = 0.0008), III (P = 0.0010), IV (P < 0.0001), V (P < 0.0001), and citrate synthase (CS) (P < 0.0001) in the intestine; complexes I (P < 0.0001), II (P < 0.0001), and CS (P = 0.0122) in the muscle; and complexes I (P < 0.0001), II (P < 0.0001), and III (P < 0.0001) in the liver. Similarly, dietary composition significantly affected complexes I (P < 0.0001), II (P < 0.0001), IV (P < 0.0001), V (P < 0.0001), and CS (P < 0.0001) in the intestine; complexes I (P < 0.0001), II (P < 0.0001), III (P = 0.0002), IV (P < 0.0001), V (P = 0.0060), and CS (P < 0.0001) in the muscle; and complexes I (P < 0.0001), II (P < 0.0001), IV (P < 0.0001), V (P < 0.0001), and CS (P < 0.0001) in the liver activities except complex III activities in intestine (P = 0.0817) and liver (P = 0.4662). The diet × temperature interaction impacted CS activity in the intestine (P = 0.0010), complex II in the muscle (P = 0.0079), and complexes I (P = 0.0009) and II (P = 0.0348) in the liver. Overall, comparing partial to full dietary substitution of FO with CO, partial dietary replacement showed similar effects on complex activities.

Diets are one of the most important consideration in aquaculture production as more than 50% of production costs are incurred in raising cultured fish to market size. As the price of FM and FO continues to increase, it is necessary to seek alternative sources of proteins and oils for sustainable aquaculture development. Plant-based ingredient sources have appeared as sustainable alternatives; however, it is uncertain whether the uses of plant-based alternatives will be appropriate in securing the production of carnivorous aquaculture species in the context of global warming, with studies postulating on the potential effects of climate change on fish growth and health. A study was conducted to examine how the replacement of FM and FO with sustainable plant-based protein and camelina oil (CO) as ingredient sources at 14 or 18 °C would affect the growth performance, nutrient utilization efficiencies, and mitochondrial enzyme activity in rainbow trout. Based on the results, mitochondrial enzyme activities were generally higher at 18 °C and CO could replace 50% dietary FO without negative effects on rainbow trout. Overall, our study demonstrated that animal-based protein with CO is as good as animal-based with FO for growth and health, thus providing potentially sustainable diet options for aquaculture.

Stress induced by crude glycerol in a thermophilic digester: microbial community divergence and resilience, but slow process recovery.

Recovery from stress is an important property for anaerobic digestion (AD). Although AD is quite adaptable with regard to waste composition, new substrates added to stable systems may cause process decline. We tested whether crude glycerol would cause stress to a thermophilic AD microbiome previously stabilized long-term on a low C/N ratio feedstock. Three-percent (v/v) crude glycerol was added to the basal substrate (poultry litter) for two hydraulic retention time (HRT) periods. This caused stress where biogas volume and methane percentage dramatically decreased and VFA levels increased. When the basal substrate was resumed, secondary inhibition occurred, resulting in even greater stress (biogas production ceased, methane 3.6%). Unassisted recovery of system processes required eight HRT periods. In contrast, crude glycerol applied at a lower organic loading rate did not cause inhibition. Crude glycerol caused changes in dominance in the microbial community (16S rRNA pyrotags). Although process resilience was slow, the recovery of digester functions occurred in conjunction with the recovery of community structure, particularly putative syntrophic acetate-oxidizing bacteria. KEY POINTS: • Crude glycerol caused stress in thermophilic co-digestion with poultry litter. • Unassisted resilience of digester functions (methane) required 8 HRT. • Syntrophic acetate-oxidizing bacteria implicated for keystone resilience functions. Graphical abstract.

Metagenome Sequences of a Thermophilic Anaerobic Digester Adapted to a Low C/N Ratio, High-Ammonia Feedstock (Poultry Litter).

We sequenced the metagenome of a pilot-scale thermophilic digester with long-term, stable performance on poultry litter feedstock which has a very low C/N ratio, a high ammonia level, and high lignocellulose content. Firmicutes were the dominant phylum (68.9%). Other abundant phyla included Bacteroidetes, Euryarchaeota, and Thermotogae This microbiome represents a hydrogenotrophic methanogenic community with high diversity.

Metagenome Sequences of Sediment from a Recovering Industrialized Appalachian River in West Virginia.

We sequenced two metagenomes from upper sediment layers (0 to 5 and 6 to 10 cm) from the Kanawha River, West Virginia. The watershed includes inputs from the forested Appalachian Mountains, surface coal mining, municipal residues, and extensive chemical manufacturing. The dominant bacterial phyla were Proteobacteria, Bacteriodetes, Firmicutes, Actinobacteria, and Chloroflexi Xenobiotic degradation pathways were present.

Biochemical methane potential of oil-extracted microalgae and glycerol in co-digestion with chicken litter.

The objective of this work was to evaluate the technical feasibility of using both oil-extracted microalgae (M) and glycerol (G) in co-digestion with chicken litter (CL), thereby improving biochemical methane potential (BMP). Different feedstock ratios of M (0-30%), G (0-3%) and CL (67-100%) were investigated to determine the best co-digestion condition under mesophilic conditions. According to the modified Gompertz model, the best BMP (131.1mLCH4gVSfed-1) was obtained with the triple co-digestion (M:G:CL) in a proportion of 30:3:67. This yielded a methane production rate (µm) of 3.3mLCH4gVSfed-1d-1 and a lag time (λ) of 17.4d. This treatment reduced chemical oxygen demand (COD) by 91.02% and increased the methane yield 15.8% with respect to the CL control.

Transcriptome Analysis of Invasive Plants in Response to Mineral Toxicity of Reclaimed Coal-Mine Soil in the Appalachian Region.

Efficient postmining reclamation requires successful revegetation. By using RNA sequencing, we evaluated the growth response of two invasive plants, goutweed (Aegopodium podagraria L.) and mugwort (Artemisia vulgaris), grown in two Appalachian acid-mine soils (MS-I and -II, pH ∼ 4.6). Although deficient in macronutrients, both soils contained high levels of plant-available Al, Fe and Mn. Both plant types showed toxicity tolerance, but metal accumulation differed by plant and site. With MS-I, Al accumulation was greater for mugwort than goutweed (385 ± 47 vs 2151 ± 251 µg g-1). Al concentration was similar between mine sites, but its accumulation in mugwort was greater with MS-I than MS-II, with no difference in accumulation by site for goutweed. An in situ approach revealed deregulation of multiple factors such as transporters, transcription factors, and metal chelators for metal uptake or exclusion. The two plant systems showed common gene expression patterns for different pathways. Both plant systems appeared to have few common heavy-metal pathway regulators addressing mineral toxicity/deficiency in both mine sites, which implies adaptability of invasive plants for efficient growth at mine sites with toxic waste. Functional genomics can be used to screen for plant adaptability, especially for reclamation and phytoremediation of contaminated soils and waters.

Application of Anaerobic Digestion Model No. 1 to describe the syntrophic acetate oxidation of poultry litter in thermophilic anaerobic digestion.

A molecular analysis found that poultry litter anaerobic digestion was dominated by hydrogenotrophic methanogens which suggests that bacterial acetate oxidation is the primary pathway in the thermophilic digestion of poultry litter. IWA Anaerobic Digestion Model No. 1 (ADM1) was modified to include the bacterial acetate oxidation process in the thermophilic anaerobic digestion (TAD). Two methods for ADM1 parameter estimation were applied: manual calibration with non-linear least squares (MC-NLLS) and an automatic calibration using differential evolution algorithms (DEA). In terms of kinetic parameters for acetate oxidizing bacteria, estimation by MC-NLLS and DEA were, respectively, km 1.12 and 3.25 ± 0.56 kg COD kg COD(-1)d(-1), KS 0.20 and 0.29 ± 0.018 kg COD m(-3) and Yac-st 0.14 and 0.10 ± 0.016 kg COD kg COD(-1). Experimental and predicted volatile fatty acids and biogas composition were in good agreement. Values of BIAS, MSE or INDEX demonstrate that both methods (MC-NLLS and DEA) increased ADM1 accuracy.

Microbial community structure of a pilot-scale thermophilic anaerobic digester treating poultry litter.

The microbial community structure of a stable pilot-scale thermophilic continuous stirred tank reactor digester stabilized on poultry litter was investigated. This 40-m(3) digester produced biogas with 57% methane, and chemical oxygen demand removal of 54%. Bacterial and archaeal diversity were examined using both cloning and pyrosequencing that targeted 16S rRNA genes. The bacterial community was dominated by phylum Firmicutes, constituting 93% of the clones and 76% of the pyrotags. Of the Firmicutes, class Clostridia (52% pyrotags) was most abundant followed by class Bacilli (13% pyrotags). The bacterial libraries identified 94 operational taxonomic units (OTUs) and pyrosequencing identified 577 OTUs at the 97% minimum similarity level. Fifteen OTUs were dominant (≥2% abundance), and nine of these were novel unclassified Firmicutes. Several of the dominant OTUs could not be classified more specifically than Clostridiales, but were most similar to plant biomass degraders, including Clostridium thermocellum. Of the rare pyrotag OTUs (<0.5% abundance), 75% were Firmicutes. The dominant methanogen was Methanothermobacter which has hydrogenotrophic metabolism, and accounted for >99% of the archaeal clones. Based on the primary methanogen, as well as digester chemistry (high VA and ammonia levels), we propose that bacterial acetate oxidation is the primary pathway in this digester for the control of acetate levels.

Thermophilic anaerobic co-digestion of poultry litter and thin stillage.

The purpose of this study was to test whether the performance of a thermophilic CSTR digester that has been stabilized on poultry litter will be enhanced or diminished by the addition of thin stillage as co-substrate. Replicate laboratory digesters, derived from a stable pilot-scale digester, were operated with increasing ratios (w/w) of thin stillage/poultry litter feedstock. After a period of adaptation to 20% and 40% thin stillage, digester performance showed increases in biogas, percent methane and COD removal, as well as a decrease in volatile acids. Peak performance occurred with 60% thin stillage. However, 80% thin stillage caused significant reduction of performance, including declines of methanogenic activity and COD removal. In conclusion, supplementing the thermophilic digestion of poultry litter with thin stillage improved the bioenergy (methane) output, but thin stillage became inhibitory at high concentrations.

Fundamentals of microbial community resistance and resilience.

Microbial communities are at the heart of all ecosystems, and yet microbial community behavior in disturbed environments remains difficult to measure and predict. Understanding the drivers of microbial community stability, including resistance (insensitivity to disturbance) and resilience (the rate of recovery after disturbance) is important for predicting community response to disturbance. Here, we provide an overview of the concepts of stability that are relevant for microbial communities. First, we highlight insights from ecology that are useful for defining and measuring stability. To determine whether general disturbance responses exist for microbial communities, we next examine representative studies from the literature that investigated community responses to press (long-term) and pulse (short-term) disturbances in a variety of habitats. Then we discuss the biological features of individual microorganisms, of microbial populations, and of microbial communities that may govern overall community stability. We conclude with thoughts about the unique insights that systems perspectives - informed by meta-omics data - may provide about microbial community stability.

Role of the 14-3-3 protein in carbon metabolism of the pathogenic yeast Candida albicans.

We previously demonstrated that the pathogenic yeast Candida albicans effectively adapts to utilize L-sorbose (Sou+) by a novel mechanism based on the loss of one copy of chromosome 5, probably due to the reduction of copy number of a negative regulator located on this chromosome. We report here another negative regulator of L-sorbose utilization, an orthologue of the Saccharomyces cerevisiae BMH1 gene, which encodes the evolutionarily conserved protein 14-3-3. This essential gene is located on chromosome 1, does not have paralogues, and is supposedly a component of the regulatory network. Experiments involving disruption of one allele of BMH1 and overexpression of BMH1 revealed that BMH1 represses the transcription of SOU1, which is responsible for the utilization of L-sorbose. Although the exact mechanism of the interaction between BMH1 and SOU1 is not known, it is clear that the control is based on the ratio of gene copy number, and that BMH1 does not control the loss of chromosome 5, the major mechanism producing Sou+ mutants. We propose that function of BMH1 as a negative regulator of SOU1 contributes to a general cellular homeostasis. This is a first report on the role of the C. albicans essential gene BMH1 as a negative regulator of the utilization of secondary carbon source in yeast, which further substantiates the involvement of 14-3-3 proteins in diverse functions.