Upcycling of recycled minerals from sewage sludge through black soldier fly larvae (Hermetia illucens): Impact on growth and mineral accumulation.

Phosphorous (P) resources are finite. Sewage sludge recyclates (SSR) are not only of interest as plant fertilizer but also as potential source of minerals in animal nutrition. However, besides P and calcium (Ca), SSR contain heavy metals. Under EU legislation, the use of SSR derivatives in animal feed is not permitted, but given the need to improve nutrient recycling, it could be an environmentally sound future mineral source. Black soldier fly larvae (BSFL) convert low-grade biomass into valuable proteins and lipids, and accumulate minerals in their body. It was hypothesized that BSFL modify and increase their mineral content in response to feeding on SSR containing substrates. The objective was to evaluate the upcycling of minerals from SSR into agri-food nutrient cycles through BSFL. Growth, nutrient and mineral composition were compared in BSFL reared either on a modified Gainesville fly diet (FD) or on FD supplemented with either 4% of biochar (FD + BCH) or 3.6% of single-superphosphate (FD + SSP) recyclate (n = 6 BSFL rearing units/group). Larval mass, mineral and nutrient concentrations and yields were determined, and the bioaccumulation factor (BAF) was calculated. The FD + SSP substrate decreased specific growth rate and crude fat of BSFL (P < 0.05) compared to FD. The FD + SSP larvae had higher Ca and P contents and yields but the BAF for Ca was lowest. The FD + BCH larvae increased Ca, iron, cadmium and lead contents compared to FD. Larvae produced on FD + SSP showed lower lead and higher arsenic concentration than on FD + BCH. Frass of FD + BCH had higher heavy metal concentration than FD + SSP and FD (P < 0.05). Except for cadmium and manganese, the larval heavy metal concentration was below the legally permitted upper concentrations for feed. In conclusion, the SSR used could enrich BSFL with Ca and P but at the expense of growth. Due to the accumulation of Cd and Mn, BSFL or products thereof can only be a component of farmed animal feed whereas in BSFL frass heavy metal concentrations remained below the upper limit authorized by EU.

Effect of a two-step fermentation method with rumen liquor on protein quality of wheat bran and rice bran to use as poultry feed.

The availability of high quality protein rich feed in many developing countries is limited as well as expensive. Low-quality agro-industrial by-products, i.e., rice bran (RB) and wheat bran (WB), are therefore used as poultry feed irrespective of their low protein content. The main objective of the present study was to improve the protein content and the amino acid profiles of these by-products through rumen liquor mixed fermentation process. A two-step fermentation of some agricultural by-products (e.g., WB and RB) was performed in a controlled environment for 3 h and 6 h. In the 1st and 2nd steps, feedstuff (brans), McDougall buffer as well as collected rumen liquor were mixed with following the proportion of 1:2:3, respectively. After fermentation, brans were dried at 100 °C in an oven. Dried sample were used to analyze the crude protein (CP) as well as amino acid (AA) content. In 1st and 2nd fermentation of the WB, CP content increased 3.3 ± 0.2% (3 h), 4.3 ± 0.2% (6 h) and 7.7 ± 0.1% (3 h), 8.5 ± 0.1% (6 h), respectively compared to control. On the other hand, RB protein content increased by 3.3 ± 0.1% (3 h), 0.8 ± 0.1% (6 h) and 7.3 ± 0.3% (3 h), 4.0 ± 0.1% (6 h) in the 1st and 2nd fermentation step, respectively compared to control. Majority of the AA increased compared to control during the 1st fermentation step for RB and WB. However, In WB, some of the AA did not show significant difference. A number of AA were decreased after the 2nd step for both RB and WB except Methionine, which increased in both steps. In 1st and 2nd steps, Methionine increased by 24.9 ± 5.1% (3 h), 25.9 ± 5.8% (6 h) for WB and 12.2 ± 3.2% (3 h), 13.0 ± 4.5% (6 h) for RB, respectively compared to control. In conclusion brans protein and amino acid quality optimization might be possible through methodical rumen liquor mixed fermentation process for better utilization as poultry diet.

Tissue-Wide Expression of Genes Related to Vitamin D Metabolism and FGF23 Signaling following Variable Phosphorus Intake in Pigs.

Calcium (Ca) and phosphorus (P) homeostasis is maintained by several regulators, including vitamin D and fibroblast growth factor 23 (FGF23), and their tissue-specific activation and signaling cascades. In this study, the tissue-wide expression of key genes linked to vitamin D metabolism (CYP2R1, CYP27A1, CYP27B1, CYP24A1, GC, VDR) and FGF23 signaling (FGF23, FGFR1-4, KL) were investigated in pigs fed conventional (trial 1) and divergent P diets (trial 2). The tissue set comprised kidney, liver, bone, lung, aorta, and gastrointestinal tract sections. Expression patterns revealed that non-renal tissues and cells (NRTC) express genes to form active vitamin D [1,25(OH)2D3] according to site-specific requirements. A low P diet resulted in higher serum calcitriol and increased CYP24A1 expression in the small intestine, indicating local suppression of vitamin D signaling. A high P diet prompted increased mRNA abundances of CYP27B1 for local vitamin D synthesis, specifically in bone. For FGF23 signaling, analyses revealed ubiquitous expression of FGFR1-4, whereas KL was expressed in a tissue-specific manner. Dietary P supply did not affect skeletal FGF23; however, FGFR4 and KL showed increased expression in bone at high P supply, suggesting regulation to balance mineralization. Specific NRTC responses influence vitamin D metabolism and P homeostasis, which should be considered for a thrifty but healthy P supply.

The Microbiological Quality of Concentrates for Horses-A Retrospective Study on Influencing Factors and Associations with Clinical Symptoms Reported by Owners or Referring Vets.

Evidence has already been provided that feed-borne mold spores and endotoxins can trigger chronic, non-infectious respiratory disease if inhaled. Furthermore, deficiencies in feed microbiology are suspected to trigger gastrointestinal and liver disorders in horses, but the connection needs further clarification. Most of the previous studies regarding horse feed hygiene focused on forage, whereas research regarding hygienic quality of concentrates is scarce. In the present study, results of reports on hygienic quality of compound feed and cereals for horses were evaluated secondarily. Results included sensory findings, and counts of aerobic bacteria, molds and yeasts determined by cultivation and lipopolysaccharide (LPS) contents. It was found that microbial counts of compound feed exceeded VDLUFA orientation values significantly more frequently than cereals (38.4 vs. 22.6%). However, average counts of bacteria, molds and yeasts were higher in cereals than in compound feeds (p < 0.0001, respectively). Mold counts in grains were significantly higher if dry matter contents were below 86% (p = 0.0201). No relation could be established between the anamnestically reported gastrointestinal disorders or elevated liver enzyme activities and microbiological deviations. Mold counts of concentrates which were suspected to cause coughing in horses were significantly higher than mold counts of control samples (3.29 vs. 2.40 log10 cfu g−1, p = 0.0313). These results indicate that hygienic status of concentrates is relevant for horse health in the respiratory tract.

The Hygienic Status of Different Forage Types for Horses-A Retrospective Study on Influencing Factors and Associations with Anamnestic Reports.

The hygienic quality of forage for horses is discussed as a potential health hazard, especially regarding respiratory diseases, colic, and hepatopathies. Therefore, the purpose of this study was to investigate the possible relations between microbiological counts, as well as endotoxin levels and disease symptoms. Data from microbiological examination reports were analyzed retrospectively, including the results of sensory examination, microbiological counts, and lipopolysaccharide contents. Sensory analysis gave an indication of deficiencies in microbiological analysis, but both methods did not give consistently equivalent results regarding the hygienic status of forage. The strongest agreements between sensory and microbiological findings were demonstrated in haylage regarding mold contamination. The influences of dry matter content on microbiological quality could be shown in haylage and hay, whereas this did not apply to straw. Deviations regarding molds and the detection of Aspergillus species occurred, especially in haylage, with values above 70% DM detected (39.6%, p=0.0021 and 47.2%, p = 0.0393). Aspergillus was detected more frequently, and average counts were higher in samples that were suspected to induce coughing in horses (p = 0.0118 and p = 0.0313, respectively). The results of the present study emphasize the importance of feed hygiene for equine respiratory health and the need for the microbiological examination of feedstuffs, since sensory analysis cannot provide an error-free prediction of microbial counts.

Associations of carotid intima media thickness with gene expression in whole blood and genetically predicted gene expression across 48 tissues.

Carotid intima media thickness (cIMT) is a biomarker of subclinical atherosclerosis and a predictor of future cardiovascular events. Identifying associations between gene expression levels and cIMT may provide insight to atherosclerosis etiology. Here, we use two approaches to identify associations between mRNA levels and cIMT: differential gene expression analysis in whole blood and S-PrediXcan. We used microarrays to measure genome-wide whole blood mRNA levels of 5647 European individuals from four studies. We examined the association of mRNA levels with cIMT adjusted for various potential confounders. Significant associations were tested for replication in three studies totaling 3943 participants. Next, we applied S-PrediXcan to summary statistics from a cIMT genome-wide association study (GWAS) of 71 128 individuals to estimate the association between genetically determined mRNA levels and cIMT and replicated these analyses using S-PrediXcan on an independent GWAS on cIMT that included 22 179 individuals from the UK Biobank. mRNA levels of TNFAIP3, CEBPD and METRNL were inversely associated with cIMT, but these associations were not significant in the replication analysis. S-PrediXcan identified associations between cIMT and genetically determined mRNA levels for 36 genes, of which six were significant in the replication analysis, including TLN2, which had not been previously reported for cIMT. There was weak correlation between our results using differential gene expression analysis and S-PrediXcan. Differential expression analysis and S-PrediXcan represent complementary approaches for the discovery of associations between phenotypes and gene expression. Using these approaches, we prioritize TNFAIP3, CEBPD, METRNL and TLN2 as new candidate genes whose differential expression might modulate cIMT.

Shift in Circulating Serum Protein Fraction (SPF) Levels of Pregnant Jennies and Nutritional Related Aspects at Early-, Mid- and Late Gestation.

A viable tool for the monitoring of the systemic condition of the pregnant jenny may be the determination of serum protein fraction (SPF) levels, including metabolic profiling. Tissue development and composition of the growing fetus requires the mother to provide adequate nutrients to its body parts and organs. In this regard, body fluid distribution and strategic molecule transportation can be screened using SPF electropherograms and analysis of intermediate metabolites. The nutritional and health status of 12 jennies (age: 5-8 years; BW at the start: 135-138 kg; Body Condition Score, BCS [1 to 5 points] = 2.25-2.50; 4th month of gestation) were monitored throughout gestation (approximate gestation period 350-356 d). All animals were pasture-fed and were offered hay ad libitum. Individual blood samples were collected within the 4th, 7th, and 10th month following conception (ultrasound scanning). Serum biochemistry, in particular, the analysis of 6 fractions of serum proteins was carried out. The significant decrease in circulating albumin in jennies from mid- to late-gestation (p < 0.001) suggests a considerable role of dietary amino acids in the synthesis of protein for fetal tissue formation as well as body fluid distribution and blood pressure control of the jenny in those stages. Moreover, α1-globulin decreased significantly in late gestation (p < 0.047), corresponding to major organ development in the terminal fetus and supported by lipid transportation in the bloodstream of the jenny. Similarly, α2-globulin decreased in late gestation (p < 0.054) as haptoglobin, an important component for the transport of free circulating hemoglobin, is likely used for fetal synthesis. Mid-gestation, appears to be a crucial moment for adequate dietary nutrient supplementation in order to prevent homeostasis perturbation of jennies, as observed in this trial.

Mineral Phosphorus Supply in Piglets Impacts the Microbial Composition and Phytate Utilization in the Large Intestine.

A sufficient supply of phosphorus (P) to pigs in livestock farming is based on the optimal use of plant-based phytate and mineral P supplements to ensure proper growth processes and bone stability. However, a high P supplementation might bear the risk of higher environmental burden due to the occurrence of excess P and phytate degradation products in manure. In this context, the intestinal microbiota is of central importance to increase P solubility, to employ non-mineral P by the enzymatic degradation of phytate, and to metabolize residual P. A feeding experiment was conducted in which piglets were fed diets with different P levels, resulting in three groups with low, medium (covering requirements), and high concentrations of available P. Samples from caecum and colon digesta were analysed for microbial composition and phytate breakdown to estimate the microbial contribution to metabolize P sources. In terms of identified operational taxonomic units (OTU), caecum and colon digesta under the three feeding schemes mainly overlap in their core microbiome. Nevertheless, different microbial families correlate with increased dietary P supply. Specifically, microbes of Desulfovibrionaceae, Pasteurellaceae, Anaerovoracaceae, and Methanobacteriaceae were found significantly differentially abundant in the large intestine across the dietary treatments. Moreover, members of the families Veillonellaceae, Selenomonadaceae, and Succinivibrionaceae might contribute to the observed phytate degradation in animals fed a low P diet. In this sense, the targeted manipulation of the intestinal microbiota by feeding measures offers possibilities for the optimization of intestinal phytate and P utilization.

Effects of increasing dietary rye levels on physicochemical characteristics of digesta and its impact on stomach emptying as well as the formation of 'doughballs' in stomachs of young pigs.

Despite quite similar contents of starch and crude fibre of wheat and rye, the unique non-starch-polysaccharide fraction of rye (e.g. high levels of arabinoxylans and fructans) might have an impact on physicochemical properties of the digesta in pigs. Forty pigs (age: 46.8 ± 5.28 days; bodyweight: 16.1 ± 4.13 kg) were divided into four treatment groups. During four weeks, the pigs received diets consisting of wheat and/or rye, barley, soy, potato protein and a mineral supplement. The sum of wheat and rye was 69% in all diets, whereby the compound feed of each group was characterized by a different ratio (%) of wheat/rye (69/0; 46/23; 23/46; 0/69, respectively). In the stomach, 'doughballs' occurred more frequently with increasing dietary rye levels (9/10; 69% rye). With higher DM content and extract-viscosity of gastric digesta, the stomach emptying tended to be retarded in rye groups. Compared to the control group (69% wheat), maximum dietary rye levels (69%) resulted in significantly higher concentrations of lactic acid in digesta of the stomach and small intestine. With increasing lactic acid concentrations, the pH tended to be lower in small intestinal digesta. With an intensified formation of lactic acid, effects against Gram-negative bacteria, for example Salmonella, can be expected. Moreover, because of higher viscosity and the retarded stomach emptying, there could be advantages of including rye in compound feeds when a longer lasting satiety is intended, for example when feeding pregnant sows (regularly fed restrictively).

Marination increases the bioavailability of lead in game meat shot with lead ammunition.

As a consequence of the toxicological lead characteristics, a reduction of its exposure should consider all sources. Game meat might contain elevated levels of lead due to the use of lead ammunition. The aim of the present study was to investigate the effects of acidic marination on the bioavailability of ammunition-derived lead in game meat (Roe deer), using the growing pig as an animal model. Furthermore, the study should provide evidence that the large-area scattering of lead particles leads to noticeable differences in the individual lead intake per game meat portion. Pigs of group A (n 7) received lead-shot game meat, which was cooked in water. Pigs of group B (n 7) received lead-shot game meat, which was first marinated (wine and vinegar) and then cooked. The lead content of both game meat preparations was equal with 0⋅77-0⋅79 mg Pb/portion. Pigs of group C (n 4) received lead-free game meat, which was also marinated and cooked. Additionally, lead acetate was administered intravenously to group D pigs (n 4). Blood samples were taken on elevated time points before and after game meat intake/i.v.-application. The acidic marination increased the bioavailability of orally ingested lead, resulting in significantly higher blood lead concentrations. The bioavailability of lead was 2⋅7 % when game meat was just cooked and 15 % when the meat was marinated before. The considerable variation of the individual blood lead concentrations suggests that an inhomogeneous distribution of ammunition-derived lead particles (in terms of size and number) causes individually non-comparable lead intakes from the consumption of game meat.

mRNA Profiles of Porcine Parathyroid Glands Following Variable Phosphorus Supplies throughout Fetal and Postnatal Life.

Knowledge of gene expression profiles reflecting functional features and specific responsiveness of parathyroid glands (PTGs) contributes to understanding mineral homeostasis and parathyroid function in healthy and diseased conditions. The study aims to reveal effector molecules driving the maintenance of phosphorus (P) homeostasis and parathyroid hormone (PTH) responsiveness to variable P supply throughout fetal and postnatal life. In this study, a long-term dietary intervention was performed by keeping pig offspring on distinct mineral P levels throughout fetal and postnatal life. Respective adaptation processes of P homeostasis were assessed in mRNA profiles of PTGs and serum minerals. RNA sequencing data and resulting molecular pathways of PTGs showed that the PTH abundance is very strictly controlled via e.g., PIN1, CaSR, MAfB, PLC and PKA signaling to regulate PTH expression, stability, and secretion. Additionally, the observed dietary effects on collagen expression indicate shifts in the ratio between connective tissue and parenchyma, thereby affecting cell-cell contacts as another line of PTH regulation. Taken together, the mRNA profiles of porcine PTGs reflect physiological responses in-vivo following variable dietary P supplies during fetal and postnatal life. The results serve to evaluate a long-term nutrition strategy with implications for improving the mineral balance in individuals with pathological disorders.

Effects of excessive or restricted phosphorus and calcium intake during early life on markers of bone architecture and composition in pigs.

Sufficient supply of pigs with calcium (Ca) and phosphorus (P) is essential for animal health and welfare during the growth period. However, the P content in animal manure is considered as a cause of massive environmental problems in soil and aquatic ecosystems. To complement previous findings, the objective of this study is the investigation of effects of a reduced and increased Ca and P supplementation on bone mineralization and bone structure compared with the current dietary recommendation. Another aim is to find possible serum markers that would allow the assessment of adequacy of P supply for bone health during growth. The result validated that the recommended Ca and P supply is sufficient, without the addition of microbial phytases. However, addition of P has no further beneficial effects on bone stability, while P supplementation below the recommended level affects bone development and growth performance. Reduced P levels have consequences for cancellous bone density and trabecular architecture. Further fine-tuning of the P supply in conjunction with an appropriate Ca supply will contribute to a reduction in P waste and associated environmental impact while maintaining animal health and welfare.

Levels of Pb and Cd in Single Feeding Stuffs and Compound Feeds for Poultry.

The assessment of heavy metals in feeds for food-producing animals is of particular importance both for animal and human health. Single feeds can be mixed and offered to animals either crushed or pelleted, and it would be recommendable to assess heavy metal content before mixing. A total of 408 feed samples for poultry were sent to our laboratory by farmers and feed manufacturers from Germany. Single feeding stuffs (cereals and legume seeds, n = 109 and n = 86, respectively) and compound mixed feeds for laying hens (n = 63), broilers (n = 83) and turkeys (n = 67) in the years 2015 to 2018 were processed for Pb and Cd content. Analyses pointed to the lowest Pb contents in cereal feeds (0.031 and 0.101 mg/kg feed). In compound feeds, values ranged between 0.089 (broilers) and 0.181 mg/kg on an as fed basis (laying hens). The highest Pb content of 0.358 mg/kg (as fed) was determined in one sample of mixed feed for turkeys. Cd levels in cereal feeds and compound feeds were similar; Cd in legume seeds turned out to be slightly higher (0.089 mg/kg feed). Results point to differences between cereals and legume seeds as single feeds; however, safe levels were found for both, as well as for compound feeds, below the maximum permitted for Pb 10 mg/kg and Cd 1 mg/kg in feed dry matter (88%).

Variation of Hematochemical Profile and Vitamin E Status in Feral Giara Horses From Free Grazing in the Wild to Hay Feeding During Captivity.

Wildlife protection and management are important priorities for landscape identity and biodiversity preservation. Feeding practices of fauna confined in facilities during temporary captivity are fundamental to support animal health and natural behavior. Appropriate provision of feedstuffs appears to be necessary to support the best practices in respect of animal species-specific natural diet. This investigation explored the variation of the metabolic profile by means of selected metabolite and respective circulating levels in a group feral Giara horses undergoing the change of the diet, moving from natural free grazing in the wild to temporary captivity. Six Giara horses (4 mares and 2 stallions; estimated age: 2.5-3 years; body weight: 163-170 kg) were captured to monitor the serological reaction to equine infectious anemia (EIA; screening at Coggins test). Animals were sheltered in a wildlife rescue center for a duration of 4 weeks, and all received the same hay-based diet (ad libitum). On 0 and 28 days of captivity, blood serum alpha-tocopherol (α-TOH) concentration was determined alongside selected metabolites (liver enzymes, total protein and fractions, cholesterol, triglycerides, and macrominerals and trace elements). Comparative feces quality and composition were also assessed. Both serum samples (0 vs. 28 days) displayed α-TOH levels below (<2 µg/mL) adequacy established for the domestic horse. Initial levels markedly (P = .020) decreased after the 4 weeks of captivity (Δ = -32.5%). Vitamin E status and ALT levels varied significantly, but serum protein fractions did not point to significant variations before and after captivity. All horses tested negative to EIA. Monitoring of vitamin E status of wild and feral herbivores may be recommendable in the context of adequate feeding practices during captivity to prevent potential deficiency or excessive depletion.

Water consumption in small mammals (dwarf rabbits, Guinea pigs and chinchillas): New data about possible influencing factors.

Information on water requirement of small mammals is sparse and the assessment of daily water consumption is an important aspect of the veterinary practice in exotic pet nutrition. Water and feed daily intake are interlaced, that way the water to feed ratio is a meaningful indication. However, various factors such as ambient temperature or the uptake of substances to be eliminated via the kidney (i.e. calcium) must be taken into account when estimation of water consumption is carried out. Moreover, behavioural abnormalities could affect water consumption, like in the case of water uptake as a substitute action. Adequate water supply for small mammals is indispensable for health reasons (prophylaxis of urolithiasis) and consequences relevant to animal welfare. For this purpose, data from different trials enrolling no. 5 Flemish Belgian (FB) and no. 5 New Zealand White (NZW) rabbits, no. 10 dwarf rabbits, no. 7 guinea pigs and no. 7 chinchilla, were obtained to assess water intake according to feed rationing. Water balance following feed rationing was seen to differ across the species of small mammals considered in these trials. The comparative approach pointed to species-dependent water intake behaviour. Due to the species-related high variation in water uptake, it is not possible to give general recommendations for water supply of small mammals. In addition, water uptake is also influenced by the water content of the food, absolute dry matter uptake and food composition. Nevertheless, an additional supply of drinking water is recommended for all species, since unsatisfactory water supply reduces dry matter intake and may lead to energy undersupply.

Reduced phosphorus intake throughout gestation and lactation of sows is mitigated by transcriptional adaptations in kidney and intestine.

BACKGROUND: The environmental impact of pig farming need to be reduced, with phosphorus (P) being of particular interest. Specified dietary regimens and management systems contribute to meet environmental concerns and reduce economic constrains. However, pregnant and lactating sows represent vulnerable individuals, whose reproductive potential and metabolic health status relies on adequate supply of macro- and micronutrients. The aim of this study was to investigate, whether sows fed with a dietary P content that is below or above current recommendations are capable to maintain mineral homeostasis during the reproduction cycle and which endogenous mechanisms are retrieved therefore in kidney and jejunum. Nulliparous gilts were fed iso-energetic diets with recommended (M), reduced (L), or high (H) amounts of mineral P supplements throughout gestation and lactation periods. Blood metabolites and hormones referring to the P homeostasis were retrieved prior to term (110 days of gestation) and at weaning (28 days of lactation). Transcriptional responses in kidney cortex and jejunal mucosa were analyzed using RNA sequencing. RESULTS: The variable dietary P content neither led to an aberration on fertility traits such as total weaned piglets nor to an effect on the weight pattern throughout gestation and lactation. Serum parameters revealed a maintained P homeostasis as reflected by unaltered inorganic P and calcium levels in L and H fed groups. The serum calcitriol levels were increased in lactating L sows. The endocrine responses to the dietary challenge were reflected at the transcriptional level. L diets led to an increase in CYP27B1 expression in the kidney compared to the H group and to an altered gene expression associated with lipid metabolism in the kidney and immune response in the jejunum. CONCLUSIONS: Our results suggest that current P requirements for gestating and lactating sows are sufficient and over supplementation of mineral P is not required. Shifts in renal and jejunal expression patterns between L and H groups indicate an affected intermediate metabolism, which long-term relevance needs to be further clarified.

Response of Fattening Rabbits with Acorns (Quercus pubescens Willd.) Combined in the Diet: First Acquaintances on Growth Performance, Carcass Traits and Perirenal Fatty Acid Profile.

The request for functional and healthy meat presents a challenge to modern animal nutritionists and rabbit meat consumption appears to increase alongside the aging population. Novel functional feeds for food-producing animals gather the interest of the scientific community and acorns appear frequently accounted among non-competitive-with-human feeding sources, above all in slow food production systems. This investigation aimed to assess the response to acorns combined in the diet of 40 fattening rabbits, in respect of growth performance, carcass characteristics and fatty acids composition in perirenal fat. A same commercial fattening diet combined or not with shredded acorns (control, CON = 0 vs. acorn combined diet, ACD = 200 g/kg feed as fed weight, respectively) was administered for six weeks to two groups of Separator rabbits, consisting of 20 animals each. No differences in feed conversion, carcass weight at slaughter and carcass yields (24 h) were found between groups at the end of the experimental feeding. Perirenal fat profile of rabbits from the ACD group pointed to significant differences in ΣPUFA content (25.1 vs. 31.6, as a percentage of total lipids, respectively, p < 0.001) and in the Σ n - 6/n - 3 ratio (5.95 vs. 2.41). In conclusion, acorns can be used as an energy source in mixed feeds for rabbits, especially in slow production systems.

Direct Delivery of Health Promoting ß-Asp-Arg Dipeptides via Stable Co-expression of Cyanophycin and the Cyanophycinase CphE241 in Tobacco Plants.

Feed supplementation with ß-arginine-aspartate dipeptides (ß-Asp-Arg DP) shows growth promoting effects in feeding trials with fish and might also be beneficial for pig and poultry farming. Currently, these DPs are generated from purified cyanophycin (CGP), with the help of the CGP-degrading enzyme cyanophycinase (CGPase). As alternative to an in vitro production, the DPs might be directly produced in feed crops. We already demonstrated that CGP can be produced in plastids of tobacco and potato, yielding up to 9.4% of the dry weight (DW). We also transiently co-expressed CGPases in the cytosol without degrading CGP in intact cells, while degradation occurs in the homogenized plant tissue. However, transient co-expression is not feasible for field-grown CGP plants, which is necessary for bulk production. In the present study, we proved that stable co-expression of the CGPase CphE241 in CGP-producing tobacco is sufficient to degrade 2.0% CGP/DW nearly completely within 3 h after homogenization of the leaves. In intact senescing leaves, CGP is partially released to the cytosol and degraded into DPs which limits the overall accumulation of CGP but not the level of the stable DPs. Even after 48 h, 54 µmol ß-Asp-Arg DP/g DW could be detected in the extract, which correspond to 1.5% DP/DW and represents 84% of the expected amount. Thus, we developed a system for the production of ß-Asp-Arg DP in field-grown plants.

Mitochondrial PCK2 Missense Variant in Shetland Sheepdogs with Paroxysmal Exercise-Induced Dyskinesia (PED).

Four female Shetland Sheepdogs with hypertonic paroxysmal dyskinesia, mainly triggered by exercise and stress, were investigated in a retrospective multi-center investigation aiming to characterize the clinical phenotype and its underlying molecular etiology. Three dogs were closely related and their pedigree suggested autosomal dominant inheritance. Laboratory diagnostic findings included mild lactic acidosis and lactaturia, mild intermittent serum creatine kinase (CK) elevation and hypoglycemia. Electrophysiological tests and magnetic resonance imaging of the brain were unremarkable. A muscle/nerve biopsy revealed a mild type II fiber predominant muscle atrophy. While treatment with phenobarbital, diazepam or levetiracetam did not alter the clinical course, treatment with a gluten-free, home-made fresh meat diet in three dogs or a tryptophan-rich, gluten-free, seafood-based diet, stress-reduction, and acetazolamide or zonisamide in the fourth dog correlated with a partial reduction in, or even a complete absence of, dystonic episodes. The genomes of two cases were sequenced and compared to 654 control genomes. The analysis revealed a case-specific missense variant, c.1658G>A or p.Arg553Gln, in the PCK2 gene encoding the mitochondrial phosphoenolpyruvate carboxykinase 2. Sanger sequencing confirmed that all four cases carried the mutant allele in a heterozygous state. The mutant allele was not found in 117 Shetland Sheepdog controls and more than 500 additionally genotyped dogs from various other breeds. The p.Arg553Gln substitution affects a highly conserved residue in close proximity to the GTP-binding site of PCK2. Taken together, we describe a new form of paroxysmal exercise-induced dyskinesia (PED) in dogs. The genetic findings suggest that PCK2:p.Arg553Gln should be further investigated as putative candidate causal variant.

Effects of 2 colostrum and subsequent milk replacer feeding intensities on methane production, rumen development, and performance in young calves.

A growing need exists for the development of practical feeding strategies to mitigate methane (CH4) emissions from cattle. Therefore, the objective of this study was to evaluate the influence of milk replacer feeding intensity (MFI) in calves on CH4 emission, rumen development, and performance. Twenty-eight female newborn Holstein calves were randomly assigned to 2 feeding groups, offered daily either 10% of the body weight (BW) in colostrum and subsequently 10% of the BW in milk replacer (MR; 10%-MR), or 12% of the BW in colostrum followed by 20% of the BW in MR (20%-MR). In wk 3, half of each feeding group was equipped with a permanent rumen cannula. Both groups were weaned at the end of wk 12. Hay and calf starter (mixture of pelleted grains) were offered from d 1 until wk 14 and 16, respectively. A total mixed ration was offered from wk 11 onward. Feed intake was measured daily and BW, anatomical measures, and rumen size weekly. Methane production and gastrointestinal passage rate were measured pre-weaning in wk 6 and 9 and post-weaning in wk 14 and 22, with additional estimation of organic matter digestibility. Rumen fluid, collected in wk 1, 2, 3, 6, 9, 14, 18, and 22, was analyzed for volatile fatty acid concentrations. Although the experimental period ended in wk 23, rumen volume of 17 calves was determined after slaughter in wk 34. Data was analyzed using ANOVA for the effects of feeding group, cannulation, and time, if applicable. Dry matter intake (DMI) of solid feed (SF) in 20%-MR animals was lower pre-weaning in wk 6 to 10 but mostly higher post-weaning. From wk 6 onward, anatomical measures and BW were greater in 20%-MR animals, and only the differences in body condition score gradually ceased post-weaning. Following the amount of SF intake, 10%-MR calves emitted more CH4 pre-weaning in wk 9, whereas post-weaning the 20%-MR group tended to have higher levels. Methane emission intensity (CH4/BW) was lower pre-weaning in 20%-MR animals but was comparable to the 10%-MR group post-weaning. Methane yield (CH4/DMI of SF) and estimated post-weaning organic matter digestibility were not affected by MFI. Rumen size normalized to heart girth was greater in 10%-MR calves from wk 5 to 10, but differences did not persist thereafter. In wk 34, rumen volume was higher in 20%-MR calves, but normalization to BW revealed no difference between feeding groups. In conclusion, high MFI reduces CH4 emission from calves pre-weaning, although this effect ceases post-weaning.