Research Institute for Farm Animal Biology (FBN)

To date, husbandry categories are defined based on the human concept of animal needs, however, to objectively assess animal welfare in defined husbandry systems, animal-related indicators must be evaluated. Here, we studied the impact of differential housing conditions on factors of the insulin-like growth factor system (IGF-system) in mother sows either housed in a conventional husbandry system corresponding to the regulations of the German husbandry form 1, or in the associated ecological facility, corresponding to the German husbandry form 4/5. Utilizing a proprietary bioassay that determines the activity of the IGF system at the level of intracellular signal transduction, we revealed altered serum IGF bioactivities in differentially housed sows. Moreover, we present a comprehensive analysis of the individual factors of the IGF system in serum of pregnant and lactating sows to elucidate which of these are modified depending on the housing conditions. An unsupervised principal component analysis suggests that IGF system factors might distinguish sows based on their housing environment. These results imply that the IGF system provides a solid basis for the documentation and objective assessment of animal health and welfare. However, a robust biomarker system for animal welfare assessment will likely require a multifactorial and integrative approach.

<ns7:p>The well-being of farmed and companion animals is increasingly recognised as integral to sustainable agroecosystems, companionship, and the One Health approach, which emphasises the interconnected health of people, animals, and the environment. The ELIXIR Domestic Animals Genome and Phenome (DAGP) Community supports genome-to-phenome analyses for farmed and companion animal species. Its aim is to coordinate, discuss, and explore the potential of data technology solutions to address key issues in animal welfare, behaviour, health, infectious diseases, metabolism, nutritional efficiency, and the preservation of genetic diversity and the environment. Through consolidating efforts to develop data standards, coordination, workflows, and visualisation, it will enhance the science underpinning rapidly growing fields in domestic animal genomics, including genome-enabled breeding, population genomics, pangenome analysis, functional genomics, genome editing, paleogenomics, phenotyping, and bio-banking. These standards will adhere to the FAIR data principles and leverage established ontologies to promote best practices in data coordination and archiving. This white paper, prepared by the ELIXIR DAGP Focus Group, summarises the current data infrastructure, resources, and tools available for domestic animal genomics and phenomics, and presents community-led plans and priorities to be implemented to meet the requirements of ELIXIR services and the animal science community. We describe how ELIXIR services can be applied in the domestic animal genomics and phenomics fields, and how we can connect projects and infrastructures that are active in the animal sciences domain. We also discuss three key priority areas: i) expanding the FAANG Data Portal for phenotype data with ELIXIR Data Platforms; ii) supporting submissions of new data types across ELIXIR Core Data Resources , including proprietary data from industry partners; and iii) strengthening connections to existing ELIXIR Communities and international consortia. This article provides a set of priorities for a Domestic Animals Genome and Phenome Community in ELIXIR and outlines the next steps to engage across stakeholders and to consolidate data for domestic animal science in Europe.</ns7:p>

Conjugated linoleic acid (CLA) is produced by bacterial biohydrogenation in the rumen of cattle, fulfills various biological functions, and is known for anti-obesity, anti-inflammation, anti-cancer, and other beneficial effects. It has numerous isomers, of which cis-9,trans-11 CLA accounts for 80% of total CLA, followed by trans-10,cis-12 CLA (t10,c12 CLA), with distinct molecular structures, oxidation efficiencies, activities, and functions. Different effects were observed, when isomers were individually supplemented in livestock nutrition. Currently, CLA is supplemented into the diets of dairy cows to improve the energy balance, and avoid negative effects of energy loss during the transition period. Furthermore, t10,c12 CLA was shown to reduce subcutaneous fat and to improve intramuscular fat (IMF) content in the carcasses of ruminants and pigs. Increasing the IMF content without increasing other fat depots and without compromising feed efficiency is an important goal in beef production. However, inconsistent and conflicting results were reported partly based on different study designs. This review aims to summarize studies on CLA supplementation in cattle, focusing on t10,c12 CLA and the effects of the dose, time, and method of supplementation on energy balance, milk yield and body composition, as well as on individual cells in vitro. This may improve our understanding of energy-saving and repartitioning effects of CLA in cattle.

IntroductionIntramuscular fat (IMF) content is a crucial factor affecting meat quality and flavor in sheep. Our previous studies demonstrated that the pyruvate dehydrogenase kinase 4 (PDK4) expression increases during adipocyte differentiation and is positively correlated with IMF content in sheep. However, the effects of the PDK4 gene on lipid metabolism in ovine adipocytes remain unclear.MethodsThe effects of PDK4 overexpression on ovine adipocyte differentiation and fat deposition were investigated. Subsequently, the key lipids and genes affected by PDK4 overexpression were identified using lipidomic and transcriptomic sequencing. Furthermore, a PDK4-knockout NIH/3T3 cell line was generated to verify the evolutionarily conserved functions of PDK4 and to examine the expression of candidate genes in a murine model.ResultsPDK4 overexpression significantly increased triglyceride deposition by 2.32-fold in ovine adipocytes (p = 0.001) but did not affect PPARγ mRNA or protein expression levels (p > 0.05). A total of 80 differentially expressed lipids (DELs) and 24 differentially expressed genes (DEGs) were identified between the overexpression (Over) and negative control (NC) groups, including 20 upregulated DELs, 60 downregulated DELs, 21 upregulated DEGs, and 3 downregulated DEGs. Additionally, PDK4 overexpression altered lipid content, composition, carbon chain length, and degree of unsaturation in ovine adipocytes. TMEM273, one of the DEGs, was negatively affected by PDK4 overexpression and closely correlated with 7 DELs (adjusted p < 0.01, |R| > 0.9). In the murine cell model, PDK4 knockout of NIH/3T3 (KO) cells significantly decreased triglyceride deposition by 0.77-fold (p < 0.01) but did not affect BSCL2 expression (p = 0.08). TMEM273 expression was significantly increased by 2.78-fold in pre-differentiated KO cells (p = 0.018) and significantly decreased by 0.83-fold in differentiated KO cells (p = 0.005).ConclusionPDK4 positively regulates fat deposition in both ovine and murine adipocytes. TMEM273 expression is negatively affected by PDK4 and correlated with lipid metabolism in sheep and mice.

The capacity for animals to produce and comprehend vocalisations that provide referential acoustic cues to their eliciting cause (e.g. predator-specific alarm calls and food calls) is a highly adaptive means of maximising the benefits of group living, and has been widely studied in diverse species. However, an underexplored dimension of referentiality is the ability to process the direction in which a vocalisation is emitted as a cue towards its referent. Here, we replicated an experimental design previously applied to dogs, chimpanzees and human infants to investigate whether domestic goats (Capra hircus) can use human voices as a directional cue in a hidden-object task. Twenty-nine goats from a UK sanctuary participated in three experimental conditions. In each condition, goats were individually presented with a human experimenter obscured by a barrier, and two identical containers, one of which was baited with food. In the reward directed speech condition, the experimenter vocalised excitedly towards the baited container while sitting closer to the un-baited container, and then the goat was able to select which container to explore. While substantial inter-individual variation existed, on average, subjects chose the baited container at above chance level across the four trials. Two control conditions explored alternative explanations for this result: a no speech condition, in which the experimenter was silent but remained in the same location as in the test condition, and a non-reward directed speech condition, in which the experimenter directed their voice away from both containers. Subjects showed no evidence of choosing the baited container at above chance level in either of these control conditions. We conclude that goats, like dogs, but not chimpanzees, are capable of attending to the directional cues provided by human voices, and discuss the possible role of domestication in the taxonomic distribution of this ability.

Fatigue is a prevalent and debilitating symptom in patients with liver cirrhosis (LC), characterized by persistent exhaustion that impairs both physical and mental function. Despite its significant impact fatigue is often overlooked in clinical practice due to its complex and subjective nature. Emerging evidence suggests central mechanisms underlying fatigue in LC, with potential links to malnutrition and sarcopenia. This study aimed to determine the prevalence of fatigue in LC patients, investigate its associations with malnutrition and sarcopenia, and identify independent indicators to aid in recognizing high-risk individuals. Additionally, the study explored the trajectory of fatigue in a structured nutritional intervention in malnourished LC patients. In a multicentric cross-sectional study, 91 LC patients and 92 matched healthy controls were recruited. Analysis focused on 56 LC patients, assessing both central and peripheral fatigue with the Fatigue Severity Scale (FSS), along with anxiety and depression (Hospital Anxiety and Depression Scale, HADS), and loneliness (De Jong Gierveld Scale). Body composition, malnutrition, and sarcopenia were evaluated using bioelectrical impedance, Global Leadership Initiative on Malnutrition (GLIM) and European Working Group on Sarcopenia in Older People (EWGSOP) II criteria. Laboratory parameters and waist circumference as indicator for ascites were assessed. Functional status was measured via handgrip strength (Jamar Dynamometer) and gait speed, and physical activity levels were assessed using the International Physical Activity Questionnaire short form (IPAQ SF). Dietary intake was evaluated with a validated Food Frequency Questionnaire (FFQ). The course of fatigue was investigated in a single-arm exploratory, intensified three-month nutritional intervention study with a 6-month follow-up in malnourished LC patients. Fatigue, affecting 68 % of LC patients, was more common than in controls and linked to both central and peripheral symptoms. LC patients also experienced higher rates of anxiety, depression, and mild loneliness. While 59 % had malnutrition and 23 % sarcopenia, neither were related to fatigue, nor anemia markers like hemoglobin or hematocrit. Fatigue was linked to increased use of proton pump inhibitors and antibiotics. Fatigue severity was correlated with increased anxiety, depression, loneliness, increased waist circumference, and lower gait speed and physical activity. Regression analysis showed increased waist circumference and reduced gait speed as independent predictors of fatigue. Nutritional intervention in malnourished LC patients led to improvements in fatigue, anxiety, and depression. Ascites and gait speed are practical clinical indicators of fatigue in LC patients. Nutritional therapy shows promise in alleviating fatigue and malnutrition in LC patients, offering a potential therapeutic approach. DRKS00021124, DRKS00021181.

Toll-like receptors (TLRs) are a class of pattern-recognition receptors that recognize pathogen- and damage-associated molecular patterns and initiate immune responses. TLRs selectively recruit distinct adapter molecules such as the myeloid differentiation primary response protein 88 (MyD88) that mediates signaling downstream of all TLRs, with the exception of TLR3. To investigate TLR signaling pathways in fish, we engineered a knockout clonal epitheliod fish cell line, named MYD88C2, using CRISPR/Cas9-mediated genome editing to disrupt the myd88 gene. We characterized the phenotype of this cell line alongside a wild-type cell line through gene-expression profiling and reporter-gene analyses in the context of stimulation with heat-killed Vibrio anguillarum, heat-killed Escherichia coli, flagellin, zymosan, and inoculation with a panel of viruses. We demonstrate that the proinflammatory response to zymosan and flagellin, as measured through the induction of proinflammatory genes, was greatly reduced in the MYD88C2 cell line. The responsiveness to zymosan was found to be partially restored by transfecting the MYD88C2 cell line with a myd88-expression plasmid. In contrast, the loss of the myd88 gene had no impact on the cytopathic effect associated with the replication of viral haemorrhagic septicemia virus (VHSV), infectious haematopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus (IPNV), spring viraemia of carp virus (SVCV) or infectious salmon anaemia virus (ISAV). These findings highlight the critical role of MyD88 in mediating specific proinflammatory responses to bacterial and fungal stimuli, while its absence has no detectable impact on viral replication or cytopathogenicity in epithelioid fish cells under the conditions tested.

Primary cell culture models are useful tools to analyze intracellular signaling pathways and interactions in detail. However, at higher passages, vital cell characteristics such as cell morphology, physiology, gene expression and cell proliferation can be compromised which may result in variable data. In the present study, we characterized cultured primary luteal cells (PLCs) and compared them to intermediate passaged luteal cells i.e. passage number 15 (P15) and high passaged luteal cells i.e. passage number 30 (P30). To explore in-vitro culture induced variabilities, PLCs were passaged repeatedly until passage number P30. Expression of cell cytoskeleton proteins was monitored by immunofluorescence and steroidogenic acute regulatory (STAR) protein expression was detected by capillary electrophoresis as physiological key parameter. The abundance of STAR, hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (HSD3B1) and luteinizing hormone/choriogonadotropin receptor (LHCGR) marker transcripts was quantified by RT-qPCR. Cell proliferation and cell viability of luteal cells were evaluated using flow cytometry. Global gene expression profiling by RNA sequencing was performed on early passaged (P3) luteal cells. Cell passaging severely reduced the expression of genes encoding marker proteins of luteal cells. Similarly, progesterone (P4) synthesis and cell proliferation were reduced significantly at higher passages. Early passaged (P3) luteal cells expressed key genes of luteal cells but with lower expression values. Luteal cells remained highly viable and consistently co-expressed both vimentin and cytokeratin-18 protein in their cytoskeleton irrespective of passage number. Together, these findings demonstrate that only short-term luteal cell primary cultures or very early passaged luteal cells (P3) are able to display molecular features that resemble that of the corpus luteum in vivo and thus are suitable in vitro models.

The objective of this study was to investigate the effects of chromium-niacin (CrNi) supplementation on production parameters, blood metabolites, antioxidant status and serum metabolite profile in transition dairy cows. A total of 28 multiparous Holstein dairy cows were included in the study. The experimental design included two treatments: a control group and a CrNi group, with each cow in the CrNi group receiving 8mg chromium and 6g niacin per day. The results showed that CrNi supplementation improved postpartum dry matter intake (DMI), which was higher in the CrNi group (21.02 vs. 19.74kg/d; P = 0.09). Milk yield was significantly increased by 1.99kg/d in the CrNi group (39.55 vs. 37.56kg/d; P = 0.04). Somatic cell count (SCC) was numerically lower in the CrNi group (217 vs. 414 × 10³ cells/mL; P = 0.08), indicating a possible improvement in udder health. Milk protein, fat, lactose and solids-not-fat percentages and yields were not significantly affected by the treatment. Body condition score (BCS) remained similar between groups. No treatment differences were found in serum calcium, BHB, NEFA, alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea, glucose, insulin, bilirubin, total protein, albumin, malondialdehyde (MDA) or total antioxidant capacity (TAC). Serum triglyceride levels were affected by CrNi (P = 0.06), with a significant treatment × time interaction (P < 0.05). Metabolomics analysis revealed that 42 metabolites were significantly altered by CrNi supplementation, with 21 metabolites upregulated, particularly those involved in lipid metabolism and microbial co-metabolism. In particular, DG (i-15:0/8:0/0:0) and Vanillic acid 4-O-sulfate were strongly upregulated, indicating increased lipid mobilization. Conversely, 21 metabolites were downregulated, including L-Threonine and Gabexate, indicating suppression of certain metabolic pathways, including anti-inflammatory processes. These results suggest that CrNi supplementation may contribute to improved production performance and metabolic health in transition dairy cows.