Protein turnover in pigs: A review of interacting factors.

Protein turnover defines the balance between two continuous and complex processes of protein metabolism, synthesis and degradation, which determine their deposition in tissues. Although the liver and intestine have been studied extensively for their important roles in protein digestion, absorption and metabolism, the study of protein metabolism has focused mainly on skeletal muscle tissue to understand the basis for its growth. Due to the high adaptability of skeletal muscle, its protein turnover is greatly affected by different internal and external factors, contributing to carcass lean-yield and animal growth. Amino acid (AA) labelling and tracking using isotope tracer methodology, together with the study of myofiber type profiling, signal transduction pathways and gene expression, has allowed the analysis of these mechanisms from different perspectives. Positive stimuli such as increased nutrient availability in the diet (e.g., AA), physical activity, the presence of certain hormones (e.g., testosterone) or a more oxidative myofiber profile in certain muscles or pig genotypes promote increased upregulation of translation and transcription-related genes, activation of mTORC1 signalling mechanisms and increased abundance of satellite cells, allowing for more efficient protein synthesis. However, fasting, animal aging, inactivity and stress, inflammation or sepsis produce the opposite effect. Deepening the understanding of modifying factors and their possible interaction may contribute to the design of optimal strategies to better control tissue growth and nutrient use (i.e., protein and AA), and thus advance the precision feeding strategy.

Changes in the Rumen Microbiota of Cows in Response to Dietary Supplementation with Nitrate, Linseed, and Saponin Alone or in Combination.

Dietary supplementation with linseed, saponins, and nitrate is a promising methane mitigation strategy in ruminant production. Here, we aimed to assess the effects of these additives on the rumen microbiota in order to understand underlying microbial mechanisms of methane abatement. Two 2-by-2 factorial design studies were conducted simultaneously, which also allowed us to make a broad-based assessment of microbial responses. Eight nonlactating cows were fed diets supplemented with linseed or saponin in order to decrease hydrogen production and nitrate to affect hydrogen consumption; also, combinations of linseed plus nitrate or saponin plus nitrate were used to explore the interaction between dietary treatments. Previous work assessed effects on methane and fermentation patterns. Rumen microbes were studied by sequencing 18S and 16S rRNA genes and ITS1 amplicons. Methanogen activity was monitored by following changes in mcrA transcript abundance. Nitrate fed alone or in combination in both studies dramatically affected the composition and structure of rumen microbiota, although impacts were more evident in one of the studies. Linseed moderately modified only bacterial community structure. Indicator operational taxonomic unit (OTU) analysis revealed that both linseed and nitrate reduced the relative abundance of hydrogen-producing Ruminococcaceae Linseed increased the proportion of bacteria known to reduce succinate to propionate, whereas nitrate supplementation increased nitrate-reducing bacteria and decreased the metabolic activity of rumen methanogens. Saponins had no effect on the microbiota. Inconsistency found between the two studies with nitrate supplementation could be explained by changes in microbial ecosystem functioning rather than changes in microbial community structure.IMPORTANCE This study aimed at identifying the microbial mechanisms of enteric methane mitigation when linseed, nitrate, and saponins were fed to nonlactating cows alone or in a combination. Hydrogen is a limiting factor in rumen methanogenesis. We hypothesized that linseed and saponins would affect hydrogen producers and nitrate would affect hydrogen consumption, leading to reduced methane production in the rumen. Contrary to what was predicted, both linseed and nitrate had a deleterious effect on hydrogen producers; linseed also redirected hydrogen consumption toward propionate production, whereas nitrate stimulated the growth of nitrate-reducing and, hence, hydrogen-consuming bacterial taxa. This novel knowledge of microbial mechanisms involved in rumen methanogenesis provides insights for the development and optimization of methane mitigation strategies.

Measurement of Methane and Ammonia Emissions from Compost-Bedded Pack Systems in Dairy Barns: Tilling Effect and Seasonal Variations.

Dairy cattle contribute to environmental harm as a source of polluting gas emissions, mainly of enteric origin, but also from manure management, which varies among housing systems. Compost-bedded pack systems use manure as bedding material, which is composted in situ daily. As current literature referring to their impact on NH3 and CH4 emissions is scarce, this study aims to characterize the emissions of these two gases originating from three barns of this system, differentiating between two emission phases: static emission and dynamic emission. In addition, the experiment differentiated emissions between winter and summer. Dynamic emission, corresponding to the time of the day when the bed is being composted, increased over 3 and 60 times the static emission of NH3 and CH4, respectively. In terms of absolute emissions, both gases presented higher emissions during summer (1.86 to 4.08 g NH3 m-2 day-1 and 1.0 to 4.75 g CH4 m-2 day-1 for winter and summer, respectively). In this way, contaminant gases produced during the tilling process of the manure, especially during the warmer periods of the year, need to be taken into account as they work as a significant factor in emissions derived from compost-bedded pack systems.

Age Evolution of Lipid Accretion Rate in Boars Selected for Lean Meat and Duroc Barrows.

Fatty acid (FA) deposition in growing-fattening pigs is mainly based on endogenous lipid synthesis, but also direct FA incorporation from the diet. To evaluate the direct fat incorporation rates and the endogenous desaturation action of the stearoyl-CoA desaturase (SCD) enzyme, a deuterium (D)-labeled saturated FA (d35-C18:0) was added to the diet. Sixteen three-way (3W) crossbred boars, and thirty-two purebred Duroc barrows homozygous for the SCD single nucleotide polymorphism rs80912566 (16 CC/16 TT), were used. Half of the animals of each genotype belonged to the growing and fattening phases. The fractional incorporation rate (FIR) of dietary fat in growing pigs was generally higher in adipose tissues, whereas in fattening pigs it was higher in the liver. Duroc pigs exhibited lower FIRs than 3W pigs, suggesting lower rates of endogenous synthesis by 3W pigs. Real fractional unsaturation rates (FURs) increased with age by the higher FIRs in 3W pigs and the de novo synthesis pathway in Duroc genotypes. Moreover, pigs carrying the SCD_T allele showed more enhanced oleic acid biosynthesis than Duroc CC pigs. In conclusion, suitable feeding protocols should be designed for each pig type to optimize production traits, considering that the metabolic pathway of FA for its deposition may differ.

The Impact of Genetics on Gut Microbiota of Growing and Fattening Pigs under Moderate N Restriction.

Characterization of intestinal microbiota is of great interest due to its relevant impact on growth, feed efficiency and pig carcass quality. Microbial composition shifts along the gut, but it also depends on the host (i.e., age, genetic background), diet composition and environmental conditions. To simultaneously study the effects of producing type (PT), production phase (PP) and dietary crude protein (CP) content on microbial populations, 20 Duroc pigs and 16 crossbred pigs (F2), belonging to growing and fattening phases, were used. Half of the pigs of each PT were fed a moderate CP restriction (2%). After sacrifice, contents of ileum, cecum and distal colon were collected for sequencing procedure. Fattening pigs presented higher microbial richness than growing pigs because of higher maturity and stability of the community. The F2 pigs showed higher bacterial alpha diversity and microbial network complexity (cecum and colon), especially in the fattening phase, while Duroc pigs tended to have higher Firmicutes/Bacteroidetes ratio in cecum segment. Lactobacillus was the predominant genus, and along with Streptococcus and Clostridium, their relative abundance decreased throughout the intestine. Although low CP diet did not alter the microbial diversity, it increased interaction network complexity. These results have revealed that the moderate CP restriction had lower impact on intestinal microbiota than PP and PT of pigs.

Annual Nitrogen Balance from Dairy Barns, Comparison between Cubicle and Compost-Bedded Pack Housing Systems in the Northeast of Spain.

The aim of this study was to determine N recovery and irreversible losses (i.e., through NH3-N volatilization) from manure in two different housing systems throughout a year using an N mass balance approach. Dietary, milk, and manure N were monitored together with outside temperatures in six dairy barns during six months, comprising two different seasons. Three barns were designed as conventional free stalls (cubicle, CUB) and the other three barns as compost-bedded packs (CB). All the barns were located in the Ebro's valley, in the northeast of Spain. Mass N balance was performed simultaneously in the six barns, during two three-month periods (Season I and II) and sampling at a 15-day interval. Results of ANOVA analysis showed that annual N retained in manure (kg/head per year) from cows housed in CUB barns was significantly higher than in manure from cows housed in CB (133.5 vs. 70.9, p < 0.001), while the opposite was observed for N losses (26.9 vs. 84.8, for CUB and CB barn, respectively; p < 0.005). The annual mean proportion of irreversible N loss from manure in relation to N intake was much lower in barns using conventional free-stall cubicles than the mean ratio registered in bedded pack systems barns.

The Impact of Producing Type and Dietary Crude Protein on Animal Performances and Microbiota Together with Greenhouse Gases Emissions in Growing Pigs.

In order to reduce dietary nitrogen and achieve an efficient protein deposition as well as decrease N wastage, we challenged the nutrient utilization efficiency of two different producing types in front of a dietary crude protein (CP) restriction and studied the role of the microbiota in such an adaptation process. Therefore, 32 pure castrated male Duroc (DU) and 32 entire male hybrid (F2) piglets were raised in a three-phase feeding regime. At each phase, two iso caloric diets differing in CP content, also known as normal protein (NP) and low protein (LP), were fed to the animals. LP diets had a fixed restriction (2%) in CP content in regards to NP ones throughout the phases of the experiment. At the end of third phase, fecal samples were collected for microbiota analysis purposes and greenhouse gases emissions, together with ammonia, were tested. No changes were found in average daily feed intake (ADFI) of animals of two producing types (Duroc vs. F2) or those consumed different experimental diets (NP vs. LP) throughout the course of study. However, at the end of each experimental phase the average body weight (BW) of hybrid animals were higher compared to Duroc pigs, whereas a reverse trend was observed for average daily gain (ADG), where Duroc pigs showed greater values with respect to hybrid ones. Despite, greater CH4 and ammonia emissions in Duroc pigs with respect to F2, no significant differences were found in contaminant gases emissions between diets. Moreover, LP diets did not alter the microbial community structure, in terms of diversity, although some genera were affected by the dietary challenge. Results suggest that the impact of reducing 2% of CP content was limited for reduction in contaminant gases emissions and highlight the hypothesis that moderate change in the dietary protein levels can be overcome by long-term adaptation of the gut microbiota. Overall, the influence of the producing type on performance and digestive microbiota composition was more pronounced than the dietary effect. However, both producing types responded differently to CP restriction. The use of fecal microbiota as biomarker for predicting feed efficiency has a great potential that should be completed with robust predictive models to achieve consistent and valid results.

Modulatory Effect of Protein and Carotene Dietary Levels on Pig gut Microbiota.

In this study we investigated the impact of dietary protein and carotene levels on microbial functions and composition during the last month of purebred fattening Duroc pigs. Fecal microbiota was characterized using 16S ribosomal RNA sequencing at two points of live, 165 (T1) and 195 (T2) days. From 70 to 165 days of age, 32 pigs were divided into two groups fed either a standard-protein (SP) or a low-protein (LP) diet. In the last month (165-195 days), all pigs received a LP diet, either carotene-enriched (CE) or not (NC). Significant differences were observed between T1 and T2 at Amplicon Sequences Variants (ASVs), phylum and genus levels. In T1 group, Prevotella, Faecalibacterium and Treponema were the genera most influenced by dietary protein, together with predicted functions related with the degradation of protein. In contrast, the CE diet did not impact the microbiome diversity, although 160 ASVs were differentially abundant between CE and NC groups at T2. Weak stability of enterotype clusters across time-points was observed as consequence of medium-term dietary interventions. Our results suggest that during the last month of fattening, dietary protein have a stronger effect than carotenes on the modulation of the compositional and functional structure of the pig microbiota.