Biomarkers for ideal protein: rabbit diet metabolomics varying key amino acids.

With the main aim of identifying biomarkers that contribute to defining the concept of ideal protein in growing rabbits under the most diverse conditions possible this work describes two different experiments. Experiment 1: 24 growing rabbits are included at 56 days of age. The rabbits are fed ad libitum one of the two experimental diets only differing in lysine levels. Experiment 2: 53 growing rabbits are included at 46 days of age, under a fasting and eating one of the five experimental diets, with identical chemical composition except for the three typically limiting amino acids (being fed commercial diets ad libitum in both experiments). Blood samples are taken for targeted and untargeted metabolomics analysis. Here we show that the metabolic phenotype undergoes alterations when animals experience a rapid dietary shift in the amino acid levels. While some of the differential metabolites can be attributed directly to changes in specific amino acids, creatinine, urea, hydroxypropionic acid and hydroxyoctadecadienoic acid are suggested as a biomarker of amino acid imbalances in growing rabbits' diets, since its changes are not attributable to a single amino acid. The fluctuations in their levels suggest intricate amino acid interactions. Consequently, we propose these metabolites as promising biomarkers for further research into the concept of the ideal protein using rabbit as a model.

A Trial Comparing Growing Rabbits Differing in 18 Generations of Selection for Growth Rate Reveals a Potential Lack of Effectiveness in the Genetic Selection Progress.

A total of 338 weaned rabbits (from the R line, selected for post-weaning growth rate) were used to evaluate the response to 18 generations of selection for increased growth rate on rabbit performance. Animals were obtained from two vitrified populations of the R line: R19V, belonging to the 18th generation (n = 165), and R37V, belonging to the 36th generation (n = 173), were allocated in individual and collective pens (178 and 160, respectively). A fattening trial was conducted from weaning (28 d of age until 63 d of age). During the trial, the body weight (BW), daily feed intake (DFI), average daily gain (ADG) and feed conversion ratio (FCR) were weekly monitored. Additionally, mortality and morbidity were daily registered. On days 49 to 53, an apparent faecal digestibility trial was also performed (12 animals per generation). Our results indicate that the generation of selection for growth rate did not affect mortality and morbidity. There were no differences in the diet digestibility according to the generation of selection. Regarding performance traits, R37V animals showed higher global BW (+6.7%; p = 0.0011) than R19V animals. R37V animals showed the same BW at weaning; however, R37V animals showed higher BW values in the last three weeks compared with R19V animals. Animals from the R37V generation also showed a higher DFI from 56 to 63 d of age (+12%; p = 0.0152) than R19V animals. However, there were no differences in global ADG and FCR between generations. These results indicate that the selection for growth rate in growing rabbits has slowed down, suggesting a lack of effectiveness in the genetic progress.

Effect of providing citrus pulp-integrated diet on fecal microbiota and serum and fecal metabolome shifts in crossbred pigs.

The study aimed to assess the impact of dehydrated citrus pulp (DCP) on growth performance, fecal characteristics, fecal bacterial composition (based on 16S rRNA analysis), and fecal and serum metabolomic profiles in crossbred pigs. 80 finishing pigs Duroc × (Landrace × Large White) were fed either a control diet (C) or a diet with 240 g/kg DCP (T) for six weeks. Including DCP in diets tended to decrease feed intake, increased (p < 0.05) the concentrations of acetic and heptanoic acids and decreased (p < 0.05) fecal butyric and branched-chain fatty acid concentrations in feces. Animals fed DCP exhibited a lower abundance of the genera Clostridium and Romboutsia, while Lachnospira significantly increased. Orthogonal partial least squares discriminant analysis plotted a clear separation of fecal and serum metabolites between groups. The main discriminant fecal metabolites were associated with bacterial protein fermentation and were downregulated in T-fed pigs. In serum, DCP supplementation upregulated metabolites related to protein and fatty acids metabolism. In conclusion, the addition of DCP as an environmentally friendly source of nutrients in pig diets, resulted in modifications of fecal bacterial composition, fermentation patterns, and overall pig metabolism, suggesting improvements in protein metabolism and gut health.

Evaluation of biotechnological processing through solid-state fermentation of oilseed cakes on extracts bioactive potential.

Oilseed cakes (OC) are natural sources of lignocellulosic biomass, produced every year in large amounts. In addition to their main applications as animal feed, plant or soil fertilizer, and compost, they present enormous potential for being used in biotechnological processes for the obtainment and extraction of valuable bioactive compounds. This work evaluated the effect of solid-state fermentation on the bioactive properties of extracts obtained from the bioprocessing of OC and evaluated the effect of solvents on the recovery of compounds with higher bioactive potential. A general decrease of EC50 values was observed for fermented extracts obtained using a mixture of water/methanol (1:1) as extraction solvent. A decrease in the minimum inhibitory concentration was observed for fermented water extracts compared to non-fermented. Additionally, growth inhibition of Listeria monocytogenes was observed when using aqueous methanolic fermented extracts. These extracts also exhibited a higher percentage of growth reduction against phytopathogenic fungi, and some extracts exhibited increased protection against genotoxic agents such as camptothecin and bisphenol A. It was demonstrated that bioprocessing of OC through SSF is an effective approach to obtaining valuable compounds with bioactive properties for use in the food, pharmaceutical or cosmetic industries.

Nutritional ecology of European rabbit (Oryctolagus cuniculus): Factors affecting chemical composition of gastric content.

Nutritional ecology seeks to unravel the extensive web of nutritional links that directs animals in their interactions with their ecological and social environments. European rabbit (Oryctolagus cuniculus) populations its endemic locations are declining and it is considered a keystone species of the Mediterranean ecosystem prompteing the interest in its conservation. The main aim of this study was to determine the nutritional composition of the diet of European rabbits through the relative and absolute chemical composition of the gastric content. To address this objective, gastric content was collected from 80 European rabbits in a Mediterranean area for the analysis of its chemical composition. To this end, gastric content was analyzed for dry matter (DM), organic matter (OM), ash, crude protein (CP), highly digestible nonnitrogenous nutrients (HDNN), neutral detergent fibre (NDF), acid detergent fibre (ADF) and lignin. The rabbits were divided into two groups: EMPTY and FULL, depending on the level of stomach filling, directly related to food intake. Our results revealed a positive correlation between the rabbits weight with DM in the gastric content, total gastric content with DM in the gastric content, and DM in gastric content with all chemical parameters analysed. The mean relative values obtained were 8.8%, 25.5%, 40.4% and 25.4%, for ash, CP, NDF and HDNN, respectively. Moreover, EMPTY rabbits had both a proportional (+19%, p = 0.002 and -40%; p = 0.004, on NDF and HDNN, respectively) and absolute (-38%, p = 0.014, -52%; p = 0.012, -52%; p = 0.011 and +83%; p = 0.008 for OM, ash, HDNN, and lignin, respectively) different proportion of nutrients in gastric contents than FULL animals. Since there is a connection between this availability and the fitness of this species, understanding the chemical composition of the rabbit's diet can be utilised to delve into its biology. Our study provides information that will help elucidate the factors affecting the chemical composition of the gastric content of European rabbits to assist land use planners and conservationists in identifying sites for conservation in Mediterranean ecosystems.

The nutritional strategy of European rabbits is affected by age and sex: Females eat more and have better nutrient optimisation.

The ecological interest in the European rabbit (Oryctolagus cuniculus) has grown since it was declared an endangered species. Nutrition is fundamental in its dispersion and the key to its success. This is the main reason why knowledge of their nutritional preferences/requirements could play a fundamental role in rabbit biology and, therefore, in their conservation. The objectives of the work will be to elucidate how age and sex affect both nutritional preferences/requirements, and the nutrient optimisation of European rabbits. To address this gap, 70 wild European rabbits were studied. The rabbits were divided according to their age: adult (n = 43) and young (n = 27). Two groups were differentiated according to the adult rabbits' sex: females (n = 28) and males (n = 15). We analysed the relative (%) and absolute (g) chemical composition of the rabbits' gastric contents: dry matter (DM), ashes (ASH), crude protein (CP), ether extract (EE), neutral detergent fibre (NDF), acid detergent fibre (ADF), and lignin, hemicelluloses and celluloses, highly digestible nonnitrogenous nutrients (HDNN: fat, starch, and soluble fibre), well from the blood was analysed with plasmatic urea nitrogen (PUN), nonesterified fatty acid (NEFA) and glucose (GLU). As a nutrient optimisation measure, the following indices between blood metabolites and stomach content were calculated: PUN/CP total content, NEFA/DM total content and GLU/DM total content ratios. Our works showed that age and sex affected the nutritional strategy of rabbits. Regarding age: adults showed lower CP (-14%; p = 0.0217) and higher HDNN (+21%; p = 0.0399) relative content than young rabbits, and absolute amount of most nutrients: DM (+59%; p = 0.001), OM (+43%; p = 0.0049), ASH (+54%; p = 0.0085), Hemicelluloses (+73%; p = 0.0084), Cellulose (+27%; p = 0.0452), and HDNN (+63%; p = 0.0012). In addition, adults showed better nutrient optimisation. Sex did not affect the relative chemical composition of the gastric content, but it showed a clear higher gastric content by females (+85%; p < 0.0001) and higher intake of most of the absolute chemical components: DM (+64%; p < 0.001), CP (+56%; p = 0.0005), OM (+58%; p = 0.0001), ASH (+44%; p = 0.0123), HDNN (+39%; p = 0.001), NDF (+59%; p = 0.001), ADF (+64%; p = 0.0003), lignin (+82%; p = 0.0036) and cellulose (+58%; p = 0.0002). Finally, we observed that females had better nutrient optimisation than males. This works supports the idea that feeding ecology and nutrition are particularly relevant to reproductive success and fitness in wild animals.

Effect of Increasing the Methionine Level and Reducing the Threonine Level in the Diet of Fast-Growing Rabbits.

The main aim of this work was to evaluate a novel combination of the first limiting amino acids (lysine, methionine, and threonine) in fast-growing rabbits (combination MHL, shown to minimise levels of plasmatic urea nitrogen in previous research, medium level of lysine, high level of methionine, and low level of threonine) and compare it with current recommendations (combination MMM, medium level of all amino acids). A total of 165 weaned rabbits (28 d old) from a paternal line selected for growth rate were used in the growth trial. The effect of the diet on apparent fecal digestibility, as well as the apparent and true ileal digestibility, were studied. Nutrient retention was also determined. Although no differences in digestibility between diets were observed, animals fed with Diet MHL improved the global average daily gain (+2.3 g/d; p = 0.0482) and feed conversion ratio (-0.10; p = 0.0229). Animals fed with Diet MMM reduced the protein-to-energy ratio retained (p = 0.0086). In conclusion, Diet MHL promoted an improvement in growth traits in a paternal line. Consequently, we propose its levels of 6.4, 5.4, and 5.0 g/kg DM of true ileal digestible lysine, methionine, and threonine, respectively.

Bioprocessing of oilseed cakes by fungi consortia: Impact of enzymes produced on antioxidants release.

Oilseed cakes (OC) present high potential as feedstock for the biobased industry. Biotechnological processes allow OC valorization by the production of diverse value-added products and simultaneously altering OC structure, improving their nutritional value, and boosting OC utilization in animal feed. This work explored the use of fungi consortium of two different species as a bioprocessing approach to improve the nutritional quality of OC, obtain enzymes and antioxidants by solid-state fermentation (SSF) of sunflower cake (SFC) and rapeseed cake (RSC). Rhyzopus oryzae and Aspergillus ibericus consortium led to the highest production of cellulase (135 U/g) and ß-glucosidase (265 U/g) while maximum protease (228 U/g) was obtained with A. niger and R. oryzae consortium. Maximum xylanase production (886 U/g) was observed in SSF of RSC resulting in high hemicellulose reduction. The synergistic action of lignocellulosic enzymes resulted in extracts with increased antioxidant potential with possible application as food additives against oxidative stress.

Unravelling the Role of Metabolites for Detecting Physiological State of Wild Animals: European Rabbit's (Oryctolagus cuniculus) Case.

European wild rabbit (Oryctolagus cuniculus) has been defined as a keystone species in the Mediterranean ecosystem. Rabbits have been classed as "endangered" by the IUCN within their native range. In this sense, animal nutrition may play a fundamental and limiting role in the conservation of wild species. The overarching goal of ecological nutrition is to unravel the extensive web of nutritional links that direct animals in their interactions with their ecological environments. The main aim of this work was to evaluate the effect of different feed intake, geographic location, animal sex, and reproductive stage on glucose, non-esterified fatty acids (NEFA), and plasmatic urea nitrogen (PUN), albumin, glutamate, and total protein metabolites. Additionally, we examined the potential of these metabolites as biomarkers. Full stomach contents and blood samples were collected from European wild rabbits (n = 89) for the analysis of the metabolites described above. Our work shows that the levels of these metabolites are affected by the sex of the animals, as well as by their reproductive stage (glucose, NEFA and albumin). There were signs of better optimisation of resources by females than by other groups of animals. These data may be interesting in the study of nutritional components that could be affecting physiological state of this species.

Nutritional Metabolites as Biomarkers of Previous Feed Intake in European Rabbit (Oryctolagus cuniculus): Applications on Conservation.

European wild rabbit (Oryctolagus cuniculus) populations have drastically reduced, and recently, rabbits have been classed as "endangered" by the IUCN. This animal plays an important ecological role in Mediterranean ecosystems and its introduction could significantly contribute to ecological restoration. Rabbits have high nutrient requirements that apparently cannot be covered in all ecosystems, and there are clues that nutrition can limit their abundance and density. On the other hand, some studies reflect the effects of food availability on the metabolomic status of other animal species, but there are no specific studies on this keystone species. The main aim of this work is to find biomarkers to assess the previous levels of ingestion of European rabbits (Oryctolagus cuniculus). To address this gap, gastric content and blood samples were collected from European rabbits (n = 99) in a Mediterranean area for the analysis of glucose, non-esterified fatty acids (NEFA), plasmatic urea nitrogen (PUN), albumin, glutamate and total protein metabolites. Depending on their previous feed intake (gastric content and the ratio between the gastric content and the weight of the animal), the animals were divided into two groups (lower and normal previous feed intake). Our work shows that the metabolomic profiles of the animals were affected. Levels of glucose (+82%; p = 0.0003), NEFA (−61%; p = 0.0040) and PUN (+139%; p < 0.001) were different in the animals with lower previous feed intake than the animals with normal previous feed intake. This work summarises that metabolic phenotype can be interesting when seeking to discover the limiting nutrients and food availability in diets that could affect the ecological fitness and conservation of European wild rabbits. It is important to mention that in this work, only the effects on six different metabolites have been analysed and more studies are necessary to complement the knowledge of possible metabolites that indicate the level of ingestion in this species and others. These (and new) biomarkers could be used as a tool to provide information about individual or population characteristics that other physiological parameters cannot detect, improving the conservation physiology field.

Urea Nitrogen Metabolite Can Contribute to Implementing the Ideal Protein Concept in Monogastric Animals.

The ideal protein concept refers to dietary protein with an amino acid profile that exactly meets an animal's requirement. Low-quality protein levels in the diet have negative implications for productive and reproductive traits, and a protein oversupply is energetically costly and leads to an excessive N excretion, with potentially negative environmental impact. Urea Nitrogen (UN), which corresponds to the amount of nitrogen in the form of urea circulating in the bloodstream, is a metabolite that has been widely used to detect amino acid imbalances and deficiencies and protein requirements. This review aims to critically analyse how UN can contribute to accurately implementing the ideal protein concept in monogastric animals, particularly in pig, poultry, and rabbit nutrition (14,000 animals from 76 published trials). About 59, 37, and 4% of trials have been conducted in pigs, poultry, and rabbits, respectively. UN level was negatively correlated to main performance traits (Pearson Correlation Coefficient [PCC] of -0.98 and -0.76, for average daily gain and feed conversion ratio, respectively), and lower UN level was related to higher milk yield and concentration. High level of UN was positively correlated to N excretion (PCC = 0.99) and negatively correlated to protein retention (PCC = -0.99). Therefore, UN in blood seems to be a proper indicator of amino acid imbalance in monogastric animals. Great variability in the use of UN was observed in the literature, including uses as determination medium (blood, plasma, or serum), units, and feeding system used (ad libitum or restricted), among others. A standardization of the methods in each of the species, with the aim to harmonize comparison among works, is suggested. After review, UN measurement in plasma and, whenever possible, the utilization of the same nutritional methodology (ad libitum conditions or restriction with blood sampling after refeeding at standardised time) are recommended. More studies are necessary to know the potential of UN and other bioindicators for amino acid deficiencies evaluation to get closer to the ideal protein concept.

The Duration of the Trial Influences the Effects of Mineral Deficiency and the Effective Phytase Dose in Broilers' Diets.

Two trials varying in duration (short- and long-term) were conducted to evaluate the effects of providing deficient (NC) or sufficient (PC) Ca and P levels, and different doses of a new phytase (250, 500, and 1000 FTU/kg feed), in broiler feed on growth performance, nutrient digestibility and retention, and tibia mineralization. A total of 80 and 490 male chicks (Ross) of 21 and 1 days of age were used in the short- and long-term trials, respectively. In the long-term trial, chicks fed NC diets showed a lower (p < 0.05) average daily gain and feed intake compared to chicks fed PC and a greater (p < 0.05) feed conversion ratio compared to 500 and 1000 FTU/kg feed during the starting period. Regarding the effects on minerals' and nutrients' coefficients of retention, animals fed NC showed a significantly higher digestibility for P than those fed the PC diet in the long-term trial. Additionally, feeding 250 to 500 FTU/kg diets increased most of the nutrients' digestibility in the short-term but only P digestibility in the long-term trial. Tibia mineralization increased linearly with phytase addition (p < 0.05) only in the long-term trial. In conclusion, the effects of dietary mineral and phytase levels on growth performance are more noticeable in young animals. In addition, the duration of the trial is key due to a possible adaptation phenomenon of birds to low P supplementary levels.

Biomarkers and De Novo Protein Design Can Improve Precise Amino Acid Nutrition in Broilers.

Precision nutrition in broilers requires tools capable of identifying amino acid imbalances individually or in groups, as well as knowledge on how more digestible proteins can be designed for innovative feeding programs adjusted to animals' dynamic requirements. This work proposes two potential tools, combining traditional nutrition with biotechnological, metabolomic, computational and protein engineering knowledge, which can contribute to improving the precise amino acid nutrition of broilers in the future: (i) the use of serum uric nitrogen content as a rapid biomarker of amino acid imbalances, and (ii) the design and modeling of de novo proteins that are fully digestible and fit exactly to the animal's requirements. Each application is illustrated with a case study. Case study 1 demonstrates that serum uric nitrogen can be a useful rapid indicator of individual or group amino acid deficiencies or imbalances when reducing dietary protein and adjusting the valine and arginine to lysine ratios in broilers. Case study 2 describes a stepwise approach to design an ideal protein, resulting in a potential amino acid sequence and structure prototype that is ideally adjusted to the requirements of the targeted animal, and is theoretically completely digestible. Both tools can open up new opportunities to form an integrated framework for precise amino acid nutrition in broilers, helping us to achieve more efficient, resilient, and sustainable production. This information can help to determine the exact ratio of amino acids that will improve the efficiency of the use of nitrogen by poultry.

Degradation of lignocellulosic matrix of oilseed cakes by solid-state fermentation: fungi screening for enzymes production and antioxidants release.

BACKGROUND: Vegetable oils are yearly produced in large amounts generating solid by-products, the oilseed cake (OC). OCs are lignocellulosic materials that have been used for animal feed with some limitations due to high fibre content from the plant cell walls. Biotechnological processes can help to overcome these limitations and contribute to up-grading such by-products, enhancing their nutritional value as feed ingredients. RESULTS: All fungal species were able to decrease neutral detergent fibre and acid detergent fibre in all by-products. Additionally, relevant enzymes were produced by the three fungi studied resulting in an improved antioxidant capacity of all fermented OCs. Aspergillus niger led to the highest activity of cellulase (109 U g-1 ), xylanase (692 U g-1 ) and protease (157 U g-1 ) per dry OC matter and to the recovery of an extract rich in antioxidants, with the highest scavenging potential of free radicals and superoxide anion, iron chelation ability and reducing power. Rhyzopus oryzae produced the highest activity of ß-glucosidase (503 U g-1 ) and led to the highest liberation of total phenolic content (TPC). Principal components analysis showed that extracts with high antioxidant potential were obtained in solid-state fermentation (SSF) with high enzymatic activity. A positive correlation was established between the action of ß-glucosidase and TPC. CONCLUSION: Within the same bioprocess it was possible to improve the nutritional value of OCs and to obtain relevant bioactive compounds such as lignocellulosic enzymes and phenolic compounds with antioxidant potential, resulting in a significant improvement of already valuable by-products with commercial interest for animal feed. © 2021 Society of Chemical Industry.

Evaluation of Phosphorus Digestibility from Monocalcium and Dicalcium Phosphate Sources and Comparison between Total Tract and Prececal Digestibility Standard Methods in Broilers.

The objective of this study was to compare the total tract (total excreta and marker) and prececal methodologies to determine phosphorus (P) digestibility and to evaluate its variation as a function of the physicochemical characteristics of the inorganic phosphate used (monocalcium, MCP and dicalcium, DCP) from different commercial sources. A total of 176 1-day-old male broilers were used in two digestibility experiments. In Experiment 1, one MCP and one DCP were incorporated in the basal diet at two levels. In Experiment 2, MCP and DCP from three commercial sources were incorporated to the basal diet at one level. Physicochemical characteristics of inorganic phosphates were examined, as well. Additionally, bone mineralization and growth performance traits were investigated in both trials. The digestibility of MCP ranged from 75.2 to 87.4% and from 80.5 to 86.6% for DCP amongst methodologies, but differences between total tract and preceal methodologies were not statistically significant. Particle size, surface area, degree of crystallinity and impurities varied amongst commercial sources. The P digestibility of the three tested commercial sources of MCP was 79.6% (MCP1), 70.2% (MCP2) and 65.6% (MCP3); p > 0.05. The P digestibility of the 3 tested commercial sources of DCP was 80.1% (DCP1), 77.4% (DCP2) and 71.4% (DCP3); p > 0.05.

Effect of Dietary Mineral Content and Phytase Dose on Nutrient Utilization, Performance, Egg Traits and Bone Mineralization in Laying Hens from 22 to 31 Weeks of Age.

A total of 192 laying hens were used to evaluate the effect of dietary mineral content and phytase dose on nutrient utilization, egg production and quality and bone mineralization of young laying hens. Four dietary treatments were studied: PC, positive control with no added phytase, 4.07% Ca and 0.61% P; NC, negative control with no added phytase, 2.97% Ca and 0.37% P; and P500 and P1000, where NC diet was supplemented with phytase at 500 and 1000 FTU/kg, respectively. Hens' performance and egg traits were controlled from 22 to 31 weeks of age. Coefficients of total tract apparent digestibility (CTTAD) of nutrients were determined at 25 and 31 weeks of age. Apparent ileal digestibility (AID) and blood content of Ca and P, as well as bone traits, were determined at 31 weeks of age. Ca and P retention was higher in birds on PC diet at 25 weeks, but not at 31 weeks of age compared to those on NC diet (p < 0.05). P1000 birds had the highest CTTAD values for dry and organic matter at both ages (p < 0.001). CTTAD of Ca was significantly higher in P1000 diet than in NC diet at 31 weeks of age (p < 0.001). Birds fed with P500 diet at 25 weeks of age and P1000 at 31 weeks of age showed higher CTTAD and retention of P, but lower excretion of P than those fed NC diet (p < 0.05). Phytase inclusion linearly increased AID of dry matter and P (p < 0.001). P500 hens fed had the greatest body weight at the end of the trial (p < 0.05) and P1000 birds had the best feed conversion ratio (p < 0.05). Fowl fed a PC diet produced eggs with higher shell thickness and yolk color than those fed on NC diet (p < 0.05). Phytase inclusion linearly increased the yolk color (p < 0.05). Tibia of laying hens fed with PC had significantly higher ash content than those on NC diet (p < 0.05), and birds fed with P1000 presented intermediate values. It can be concluded that it would be advisable to increase the dose of phytase in the feed of laying hens to obtain long-term benefits.

Do Growing Rabbits with a High Growth Rate Require Diets with High Levels of Essential Amino Acids? A Choice-Feeding Trial.

As a consequence of the genetic selection process in growing rabbits, there are currently fast-growing animals exhibiting an average daily gain that may exceed 45 g/d. The protein requirements of these animals, namely amino acid requirements, may differ from animals with low growth rates. The objective of this work was to evaluate growth performance, the coefficient of total tract apparent digestibility (CTTAD), the apparent ileal digestibility (AID) of amino acids and nutrient retention of fast-growing rabbits when they had access to a diet with high levels of amino acids and/or a diet formulated with current nutritional recommendations in a choice-feeding trial. To this end, two diets were formulated: the M diet following current nutritional recommendations for growing rabbits (including 8.1, 5.8 and 6.9 g/kg dry matter (DM) of total lysine, sulphur amino acids and threonine, respectively) and the H diet with higher levels of total lysine, sulphur amino acids and threonine (9.4, 6.6 and 7.8 g/kg DM, respectively). A total of 220 weaned rabbits, from a paternal line selected for the growth rate, had free access to the M diet, the H diet or both (MH) diets from 28 to 63 days of age. The CTTAD of DM, crude protein and gross energy from 49 to 53 days of age as well as the AID of amino acids of the diets at 63 days of age were determined. Protein, amino acids and energy retained in the empty body from 28 to 63 days of age were also registered. No significant differences in growth performance, CTTAD, AID and nutrient retention between dietary treatments were observed. However, animals fed the H diet showed a higher AID of cysteine (p < 0.05) and higher threonine retention (p < 0.05) compared to the M diet. As regards the choice-feeding trial, MH animals showed a higher feed intake of the M diet compared to the H diet (+8.61%; p < 0.001), and furthermore, more than 50% of the animals preferred the M diet throughout the experimental period (p < 0.05). Our results suggest that animals with high growth rates do not show significantly higher productive traits when fed the H diet compared to the M diet. As regards choice feeding, MH animals were capable of choosing their preferred diet, showing high intra-individual repeatability in preference for the M diet. It would be interesting to continue studying this behaviour of choice based on amino acid levels.

Comparative performance of three sampling techniques to detect airborne Salmonella species in poultry farms.

Sampling techniques to detect airborne Salmonella species (spp.) in two pilot scale broiler houses were compared. Broilers were inoculated at seven days of age with a marked strain of Salmonella enteritidis. The rearing cycle lasted 42 days during the summer. Airborne Salmonella spp. were sampled weekly using impaction, gravitational settling, and impingement techniques. Additionally, Salmonella spp. were sampled on feeders, drinkers, walls, and in the litter. Environmental conditions (temperature, relative humidity, and airborne particulate matter (PM) concentration) were monitored during the rearing cycle. The presence of Salmonella spp. was determined by culture-dependent and molecular methods. No cultivable Salmonella spp. were recovered from the poultry houses' surfaces, the litter, or the air before inoculation. After inoculation, cultivable Salmonella spp. were recovered from the surfaces and in the litter. Airborne cultivable Salmonella spp. Were detected using impaction and gravitational settling one or two weeks after the detection of Salmonella spp. in the litter. No cultivable Salmonella spp. were recovered using impingement based on culture-dependent techniques. At low airborne concentrations, the use of impingement for the quantification or detection of cultivable airborne Salmonella spp. is not recommended. In these cases, a combination of culture-dependent and culture-independent methods is recommended. These data are valuable to improve current measures to control the transmission of pathogens in livestock environments and for optimising the sampling and detection of airborne Salmonella spp. in practical conditions.

The use of agricultural substrates to improve methane yield in anaerobic co-digestion with pig slurry: effect of substrate type and inclusion level.

Anaerobic co-digestion of pig slurry with four agricultural substrates (tomato, pepper, persimmon and peach) was investigated. Each agricultural substrate was tested in co-digestion with pig slurry at four inclusion levels: 0%, 15%, 30% and 50%. Inclusion levels consisted in the replacement of the volatile solids (VS) from the pig slurry with the VS from the agricultural substrate. The effect of substrate type and inclusion level on the biochemical methane potential (BMP) was evaluated in a batch assay performed at 35 °C for 100 days. Agricultural substrate's chemical composition was also analyzed and related with BMP. Additionally, Bacteria and Archaea domains together with the four main methanogenic archaeal orders were quantified using quantitative real-time TaqMan polymerase chain reaction (qPCR) at the end of the experiment to determine the influence of agricultural substrate on sludge's microbial composition. Results showed that vegetable substrates (pepper and tomato) had higher lipid and protein content and lower carbohydrates than fruit substrates (persimmon and peach). Among substrates, vegetable substrates showed higher BMP than fruit substrates. Higher BMP values were obtained with increasing addition of agricultural substrate. The replacement of 50% of VS from pig slurry by tomato and pepper increased BMP in 41% and 44%, respectively compared with pig slurry only. Lower increments in BMP were achieved with lower inclusion levels. Results from qPCR showed that total bacteria and total archaea gene concentrations were similar in all combinations tested. Methanomicrobiales gene concentrations dominated over the rest of individual archaeal orders.

Airborne particulate matter from livestock production systems: a review of an air pollution problem.

Livestock housing is an important source of emissions of particulate matter (PM). High concentrations of PM can threaten the environment, as well as the health and welfare of humans and animals. Particulate matter in livestock houses is mainly coarse, primary in origin, and organic; it can adsorb and contain gases, odorous compounds, and micro-organisms, which can enhance its biological effect. Levels of PM in livestock houses are high, influenced by kind of housing and feeding, animal type, and environmental factors. Improved knowledge on particle morphology, primarily size, composition, levels, and the factors influencing these can be useful to identify and quantify sources of PM more accurately, to evaluate their effects, and to propose adequate abatement strategies in livestock houses. This paper reviews the state-of-the-art of PM in and from livestock production systems. Future research to characterize and control PM in livestock houses is discussed.