Institut für Nutztierwissenschaften

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Investigating the interplay of stressors and health in horses through fecal cortisol metabolite analysis
(2025) Nowak, Aurelia C.; Macho-Maschler, Sabine; Biermann, Nora M.; Palme, Rupert; Dengler, Franziska; Nowak, Aurelia C.; Department of Biological Sciences and Pathobiology, Institute of Physiology and Pathophysiology, University of Veterinary Medicine Vienna, Vienna, Austria; Macho-Maschler, Sabine; Department of Biological Sciences and Pathobiology, Experimental Endocrinology, University of Veterinary Medicine Vienna, Vienna, Austria; Biermann, Nora M.; Department of Small Animals and Horses, Clinical Unit of Equine Surgery, University of Veterinary Medicine Vienna, Vienna, Austria; Palme, Rupert; Department of Biological Sciences and Pathobiology, Experimental Endocrinology, University of Veterinary Medicine Vienna, Vienna, Austria; Dengler, Franziska; Department of Biological Sciences and Pathobiology, Institute of Physiology and Pathophysiology, University of Veterinary Medicine Vienna, Vienna, Austria
Introduction: Horses are highly sensitive to stress, which can affect their wellbeing and lead to various health issues. Effective and objective stress assessment is therefore crucial for improving their care and management. The production of the glucocorticoid cortisol is increased in response to stressful stimuli and its metabolites can be measured non-invasively in feces. Therefore, this study aimed to explore the impact of different environmental and physiological stressors on fecal cortisol metabolite (FCM) concentrations in horses, with a particular focus on the relationship between stress, health, and welfare. We hypothesized that increased FCM levels may be indicative of disease and thus improve early detection and subsequent intervention. Methods: Fresh fecal samples of N = 41 horses (20 geldings and 21 mares) from the same herd were collected once weekly for 1 year. Horses had been housed in the same stable for at least a month and were accustomed to the habitat, daily routine, and social groups. Environmental conditions, health data, and potentially stressful events were recorded. Fecal concentrations of 11,17-dioxoandrostanes were measured via 11-oxoetiocholanolone enzyme immunoassay. Results: We showed stable baseline FCM concentrations of 6.3 ng/g feces (range 0.6–28.1 ng/g feces). During the summer months, median FCM concentrations increased significantly ( p < 0.05; One Way RM ANOVA), and this increase correlated with higher ambient temperatures ( p < 0.0001, adj r 2 = 0.669, Pearson Product Moment correlation). Additionally, other factors such as breed, coat color, and housing conditions influenced the FCM concentrations. Stressful events, such as riding exams and some painful conditions, also resulted in elevated FCM levels, although the magnitude of these responses varied across individual horses. However, not all diseases were associated with increased FCMs. Discussion: Our findings emphasize the complexity of the hypothalamic-pituitary-adrenal axis in horses, suggesting that while high FCM levels can indicate stress, they may not be reliable biomarkers for early disease detection. Particularly in the light of climate change, the impact of heat stress in the summer months should not be neglected and measures to improve the housing conditions accordingly should become an essential part of equine health management.
Host metabolome and faecal microbiome shows potential interactions impacted by age and weaning times in calves
(2023) Amin, Nida; Schwarzkopf, Sarah; Tröscher-Mußotter, Johanna; Camarinha-Silva, Amélia; Dänicke, Sven; Huber, Korinna; Frahm, Jana; Seifert, Jana; Amin, Nida; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Schwarzkopf, Sarah; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Tröscher-Mußotter, Johanna; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Camarinha-Silva, Amélia; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Dänicke, Sven; Institute of Animal Nutrition, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Brunswick, Germany; Huber, Korinna; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Frahm, Jana; Institute of Animal Nutrition, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Brunswick, Germany; Seifert, Jana; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany
Background: Calves undergo nutritional, metabolic, and behavioural changes from birth to the entire weaning period. An appropriate selection of weaning age is essential to reduce the negative effects caused by weaning-related dietary transitions. This study monitored the faecal microbiome and plasma metabolome of 59 female Holstein calves during different developmental stages and weaning times (early vs. late) and identified the potential associations of the measured parameters over an experimental period of 140 days. Results: A progressive development of the microbiome and metabolome was observed with significant differences according to the weaning groups (weaned at 7 or 17 weeks of age). Faecal samples of young calves were dominated by bifidobacterial and lactobacilli species, while their respective plasma samples showed high concentrations of amino acids (AAs) and biogenic amines (BAs). However, as the calves matured, the abundances of potential fiber-degrading bacteria and the plasma concentrations of sphingomyelins (SMs), few BAs and acylcarnitines (ACs) were increased. Early-weaning at 7 weeks significantly restructured the microbiome towards potential fiber-degrading bacteria and decreased plasma concentrations of most of the AAs and SMs, few BAs and ACs compared to the late-weaning event. Strong associations between faecal microbes, plasma metabolites and calf growth parameters were observed during days 42–98, where the abundances of Bacteroides, Parabacteroides , and Blautia were positively correlated with the plasma concentrations of AAs, BAs and SMs as well as the live weight gain or average daily gain in calves. Conclusion: The present study reported that weaning at 17 weeks of age was beneficial due to higher growth rate of late-weaned calves during days 42–98 and a quick adaptability of microbiota to weaning-related dietary changes during day 112, suggesting an age-dependent maturation of the gastrointestinal tract. However, the respective plasma samples of late-weaned calves contained several metabolites with differential concentrations to the early-weaned group, suggesting a less abrupt but more-persistent effect of dietary changes on host metabolome compared to the microbiome.
Selection index theory for populations under directional and stabilizing selection
(2023) Wellmann, Robin; Wellmann, Robin; Department of Animal Genetics and Breeding, University of Hohenheim, Stuttgart, Germany
Background: The purpose of a selection index is that its use to select animals for breeding maximizes the profit of a breed in future generations. The profit of a breed is in general a quantity that predicts the satisfaction of future owners with their breed, and the satisfaction of the consumers with the products that are produced by the breed. Many traits, such as conformation traits and product quality traits have intermediate optima. Traditional selection index theory applies only to directional selection and cannot achieve any further improvement once the trait means have reached their optima. A well-founded theory is needed that extends the established selection index theory to cover directional as well as stabilizing selection as limiting cases, and that can be applied to maximize the profit of a breed in both situations. Results: The optimum selection index shifts the trait means towards the optima and, in the case of stabilizing selection, decreases the phenotypic variance, which causes the phenotypes to be closer to the optimum. The optimum index depends not only on the breeding values, but also on the squared breeding values, the allele contents of major quantitative trait loci (QTL), the QTL heterozygosities, the inbreeding coefficient of the animal, and the kinship of the animal with the population. Conclusion: The optimum selection index drives the alleles of major QTL to fixation when the trait mean approaches the optimum because decreasing the phenotypic variance shifts the trait values closer to the optimum, which increases the profit of the breed. The index weight on the kinship coefficient balances the increased genetic gain that can be achieved in future generations by outcrossing, and the increased genetic gain that can be achieved under stabilizing selection by reducing the phenotypic variance. In a model with dominance variance, it can also account for the effect of inbreeding depression. The combining ability between potential mating partners, which predicts the total merit of their offspring, could become an important parameter for mate allocation that could be used to further shift the phenotypes towards their optimum values.
Transcriptional responses to diets without mineral phosphorus supplementation in the jejunum of two high-yielding laying hen strains
(2024) Abitew, Yosef A.; Reyer, Henry; Hadlich, Frieder; Oster, Michael; Trakooljul, Nares; Sommerfeld, Vera; Rodehutscord, Markus; Wimmers, Klaus; Ponsuksili, Siriluck
Phosphorus (P) is an essential mineral for all forms of life including laying hens, playing a crucial role in growth and efficient egg production. Recent studies suggest that current P recommendations might exceed the physiological demand, leading to unnecessarily high P excretions. This study on Lohmann Brown (LB) and Lohmann Selected Leghorn (LSL) laying hens (n=80; 10 replicates per strain, production period, and dietary group) investigates transcriptional changes in the jejunum, a critical intestinal segment for mineral absorption, in response to a diet either without (P-) or with (P+) a mineral supplement from monocalcium phosphate, administered over a 4-week period during the transition (15–19 weeks) or onset of laying (20–24 weeks). DESeq2 analysis of RNA sequencing data revealed that most differentially expressed genes (DEGs) varied between strains and age groups, with less pronounced effects from dietary mineral P content. The 19-week-old LB hens showed a stronger response to dietary mineral P removal, with transcripts affiliated with increased adaptation of the metabolism and decreased immune pathway activation. The identified pathways such as folate biosynthesis and p53 signaling, potentially link altered energy and amino acid metabolism (2-oxocarboxylic acid and arginine). Interestingly, genes involved in calcium transport (CALB1) and cellular signaling (PRKCA, STEAP4) along with tight junctions (CLDN2) were affected by complete removal of mineral P supplements, suggesting a promoted intestinal mineral uptake. Transcriptional regulation in the jejunum in response to low dietary mineral content is strain-specific when the laying phase begins, which may contribute to a physiological Ca:P ratio.
Functionality of the Na+-translocating NADH:quinone oxidoreductase and quinol:fumarate reductase from Prevotella bryantii inferred from homology modeling
(2024) Hau, Jann-Louis; Schleicher, Lena; Herdan, Sebastian; Simon, Jörg; Seifert, Jana; Fritz, Günter; Steuber, Julia; Hau, Jann-Louis; Institute of Biology, University of Hohenheim, Garbenstraße 30, 70599, Stuttgart, Germany; Schleicher, Lena; Institute of Biology, University of Hohenheim, Garbenstraße 30, 70599, Stuttgart, Germany; Herdan, Sebastian; Institute of Biology, University of Hohenheim, Garbenstraße 30, 70599, Stuttgart, Germany; Simon, Jörg; Microbial Energy Conservation and Biotechnology, Department of Biology, Technical University of Darmstadt, Schnittspahnstraße 10, 64287, Darmstadt, Germany; Seifert, Jana; HoLMiR-Hohenheim Center for Livestock Microbiome Research, University of Hohenheim, Leonore-Blosser-Reisen-Weg 3, 70599, Stuttgart, Germany; Fritz, Günter; Institute of Biology, University of Hohenheim, Garbenstraße 30, 70599, Stuttgart, Germany; Steuber, Julia; Institute of Biology, University of Hohenheim, Garbenstraße 30, 70599, Stuttgart, Germany
Members of the family Prevotellaceae are Gram-negative, obligate anaerobic bacteria found in animal and human microbiota. In Prevotella bryantii , the Na + -translocating NADH:quinone oxidoreductase (NQR) and quinol:fumarate reductase (QFR) interact using menaquinone as electron carrier, catalyzing NADH:fumarate oxidoreduction. P. bryantii NQR establishes a sodium-motive force, whereas P. bryantii QFR does not contribute to membrane energization. To elucidate the possible mode of function, we present 3D structural models of NQR and QFR from P. bryantii to predict cofactor-binding sites, electron transfer routes and interaction with substrates. Molecular docking reveals the proposed mode of menaquinone binding to the quinone site of subunit NqrB of P. bryantii NQR. A comparison of the 3D model of P. bryantii QFR with experimentally determined structures suggests alternative pathways for transmembrane proton transport in this type of QFR . Our findings are relevant for NADH-dependent succinate formation in anaerobic bacteria which operate both NQR and QFR.
Metabolic adaptation to energetic demands of early lactation in Holstein Friesian and Simmental cows
(2025) Ruda, Lena; Straub, C.; Scholz, A.M.; Huber, Korinna
Holstein Friesian (HF) is the highest-performing dairy breed worldwide. However, this breed is prone to suffer from health disorders and infertility due to metabolic instability. The aim of the study was to use the metabolic differences between to breed, German HF and German Simmental (SI), for discussion of the biological impact of well-known and novel pathways and their respective indicators. Therefore, a targeted metabolomics approach combined with an examination of classical clinical chemistry using plasma samples was used to describe the metabolic status 42 days before expected calving and at day 21 of lactation. In total, 27 multiparous cows (HF n = 17, SI n = 10) were enrolled in the study. All cows were kept on the same farm under identical management, feeding and housing conditions. BW of HF cows dropped while SI cows maintained their BW but produced less milk than HF cows during the first weeks of lactation. Differences were detected for the following indicators and related pathways, and were used to assess their biological impact. Branched-chain amino acids such as leucine and isoleucine at day +21 remained as high as at day −42 in HF cows; however, in SI cows, a clear drop in these amino acids was observed indicating less body protein mobilisation in this breed. Alpha amino-adipic acid and kynurenine as markers for oxidative stress balance were increased at day +21 in HF cows, only reflecting a metabolically more tensed condition in HF cows. As a conclusion, these findings confirm the impact of these metabolites on a higher risk to suffer from metabolic disorders.
Growth performance, phytate degradation, and tibia characteristics of broiler chickens fed pelleted diets with varying feed particle sizes and phytase levels
(2025) Wolfrum, Stephanie; Francesch, Maria; Jimenez-Moya, Beatriz; Feuerstein, Dieter; Rodehutscord, Markus
The objective was to investigate the effects of feed particle size and microbial phytase supplementation of pelleted diets on growth performance, myo-inositol hexakisphosphate (InsP6) degradation, and tibia characteristics in broiler chickens. Ross 308 broilers were housed in 56 floor pens in groups of 42, and seven pens were allocated to one of eight dietary treatments to measure the performance data on day 38 of age. From day 21 to 26, six birds from each floor pen were moved to metabolism units with the same distribution of treatments for digestibility and bone measurements. For the starter phase, two diets with fine or coarse particle size (429 or 657 µm mean particle size, determined after feed processing by wet sieving) were formulated and added with 500 FTU phytase/kg. For the grower and finisher phase, diets were arranged with two particle sizes (fine and coarse, 434 or 729 µm) and four phytase levels (0, 300, 600, and 1,200 FTU/kg). No significant interactions between particle size and phytase were determined. Prececal InsP6 disappearance and P digestibility were higher with fine than coarse particles, indicating that coarse grinding of rapeseed meal may not benefit birds. Increasing phytase supplementation increased ADG, ADFI, prececal InsP6 disappearance and P digestibility, tibia ash, and tibia breaking strength. Under the conditions of this study, birds were sufficiently supplied with P at a total P level of 4.0 g/kg and phytase supplementation of 1,200 FTU/kg. Renouncement of feed phosphate in the grower and finisher phase was possible without impaired performance.
Interactions between dietary phytate concentration and phytase level on phytate degradation and amino acid digestibility in broiler chickens
(2025) Wolfrum, Stephanie; Siegert, Wolfgang; Rubio-Cervantes, Ismael; Liebhold, Tina Marie; Feuerstein, Dieter; Camarinha-Silva, Amélia; Rodehutscord, Markus
The objective of the present study was to investigate the effects of dietary myo-inositol hexakisphosphate (InsP6) concentration and added phytase on gastrointestinal InsP6 degradation, prececal digestibility of P, Ca, and amino acids (AA), and MEN in broiler chickens. Dietary InsP6 was increased by graded substitution of corn starch with a mixture of 50 % soybean meal, 20 % rapeseed meal, 20 % sunflower meal, and 10 % rice bran (oilseed meal-rice bran level (ORL)). The experiment was arranged in a 4 × 3-factorial design with 4 ORL (leading to 1.4, 1.9, 2.4, and 3.0 g InsP6-P/kg) and 3 phytase levels (500, 1,500, and 3,000 FTU/kg). Male Ross 308 broilers were allocated to 84 metabolism units in groups of 10 and assigned to 1 of the 12 diets. InsP6 disappearance in the crop decreased with increasing ORL (45 to 24 %; P = 0.001). Prececal InsP6 disappearance and P digestibility linearly decreased with increasing ORL at 500 FTU/kg (83 to 56 % and 80 to 62 %; P < 0.001). Minor differences were determined for prececal InsP6 disappearance and P digestibility among ORL at 1,500 and 3,000 FTU/kg, but values decreased with increasing ORL (91 to 83 % and 87 to 81 %, respectively; P < 0.001). When prececal InsP6 disappearance relative to FTU was regressed against dietary InsP6, the relationship was non-linear at 500 FTU/kg but linear at 1,500 and 3,000 FTU/kg. Cecal InsP6 concentration increased with ORL and decreased with phytase (P < 0.001). Except for cysteine, prececal digestibility of all AA and MEN decreased with increasing ORL. The data indicated that phytase supplemented at or above 1,500 FTU/kg did not limit gastrointestinal InsP6 degradation and AA digestibility at high InsP6 concentrations of the fee
Mendelian randomisation to uncover causal associations between conformation, metabolism, and production as potential exposure to reproduction in German Holstein dairy cattle
(2025) Schwarz, Leopold; Heise, Johannes; Liu, Zengting; Bennewitz, Jörn; Thaller, Georg; Tetens, Jens; Schwarz, Leopold; Department of Animal Sciences, Georg-August-University, 37077, Göttingen, Germany; Heise, Johannes; Vereinigte Informationssysteme Tierhaltung w.V. (VIT), 27283, Verden, Germany; Liu, Zengting; Vereinigte Informationssysteme Tierhaltung w.V. (VIT), 27283, Verden, Germany; Bennewitz, Jörn; Institute of Animal Science, University of Hohenheim, 70599, Stuttgart, Germany; Thaller, Georg; Institute of Animal Breeding and Husbandry, Christian-Albrechts-University, 24118, Kiel, Germany; Tetens, Jens; Department of Animal Sciences, Georg-August-University, 37077, Göttingen, Germany
Background: Reproduction is vital to welfare, health, and economics in animal husbandry and breeding. Health and reproduction are increasingly being considered because of the observed genetic correlations between reproduction, health, conformation, and performance traits in dairy cattle. Understanding the detailed genetic architecture underlying these traits would represent a major step in comprehending their interplay. Identifying known, putative or novel associations in genomics could improve animal health, welfare, and performance while allowing further adjustments in animal breeding. Results: We conducted genome-wide association studies for 25 different traits belonging to four different complexes, namely reproduction (n = 13), conformation (n = 6), production (n = 3), and metabolism (n = 3), using a cohort of over 235,000 dairy cows. As a result, we identified genome-wide significant signals for all the studied traits. The obtained summary statistics collected served as the input for a Mendelian randomisation approach (GSMR) to infer causal associations between putative exposure and reproduction traits. The study considered conformation, production, and metabolism as exposure and reproduction as outcome. A range of 139 to 252 genome-wide significant SNPs per combination were identified as instrumental variables (IVs). Out of 156 trait combinations, 135 demonstrated statistically significant effects, thereby enabling the identification of the responsible IVs. Combinations of traits related to metabolism (38 out of 39), conformation (68 out of 78), or production (29 out of 39) were found to have significant effects on reproduction. These relationships were partially non-linear. Moreover, a separate variance component estimation supported these findings, strongly correlating with the GSMR results and offering suggestions for improvement. Downstream analyses of selected representative traits per complex resulted in identifying and investigating potential physiological mechanisms. Notably, we identified both trait-specific SNPs and genes that appeared to influence specific traits per complex, as well as more general SNPs that were common between exposure and outcome traits. Conclusions: Our study confirms the known genetic associations between reproduction traits and the three complexes tested. It provides new insights into causality, indicating a non-linear relationship between conformation and reproduction. In addition, the downstream analyses have identified several clustered genes that may mediate this association.
Weaning age influences indicators of rumen function and development in female Holstein calves
(2022) Schwarzkopf, Sarah; Kinoshita, Asako; Hüther, Liane; Salm, Laurenz; Kehraus, Saskia; Südekum, Karl-Heinz; Huber, Korinna; Dänicke, Sven; Frahm, Jana; Schwarzkopf, Sarah; Department of Functional Anatomy of Livestock, Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Kinoshita, Asako; Department of Functional Anatomy of Livestock, Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Hüther, Liane; Institute of Animal Nutrition, Friedrich-Loeffler-Institute, Braunschweig, Germany; Salm, Laurenz; Institute of Animal Nutrition, Friedrich-Loeffler-Institute, Braunschweig, Germany; Kehraus, Saskia; Institute of Animal Science, University of Bonn, Bonn, Germany; Südekum, Karl-Heinz; Institute of Animal Science, University of Bonn, Bonn, Germany; Huber, Korinna; Department of Functional Anatomy of Livestock, Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Dänicke, Sven; Institute of Animal Nutrition, Friedrich-Loeffler-Institute, Braunschweig, Germany; Frahm, Jana; Institute of Animal Nutrition, Friedrich-Loeffler-Institute, Braunschweig, Germany
Background: Prenatal and postnatal conditions are crucial for the development of calves. Primiparous cows are still maturing during pregnancy, thus competing with the nutritional needs of their offspring. Therefore, mature cows might provide a superior intrauterine condition. Furthermore, weaning calves at an older age might affect them positively as well by reducing stress and offering time for various organs and their functions to develop. We aimed to evaluate effects of mothers’ parity and calves’ weaning age on gastrointestinal development and corresponding acid–base balance. Fifty-nine female German Holstein calves (about 8 days old) were investigated in a 2 × 2 factorial experiment with factors weaning age (7 vs. 17 weeks) and parity of mother (primiparous vs. multiparous). Calves were randomly assigned to one of these four groups. Animal behavior that was observed included resting, chewing and active behavior. Results: Behavioral patterns were interactively affected by time and weaning age. Rumen sounds per 2 min increased in early-weaned calves during their weaning period. In late-weaned calves a consistently increase in rumen sounds was already recorded before their weaning period. Urinary N-containing compounds (creatinine, hippuric acid, uric acid, urea, allantoin) were interactively affected by time and weaning age. Concentrations of all measured compounds except urea increased during early weaning. All except hippuric acid concentration decreased in early-weaned calves after weaning. In late-weaned calves allantoin and uric acid increased before weaning and did not change during weaning. Conclusion: These results suggest that late-weaned calves developed adequate rumen functions and acid–base balance, whereas early-weaned calves might have suffered from ruminal acidosis and catabolism. Weaning calves at 7 weeks of age might be too early for an adequate rumen development.
Unraveling the role of shrimp hydrolysate as a food supplement in the immune function and fecal microbiota of beagle dogs
(2025) Guilherme-Fernandes, Joana; Barroso, Carolina; Correia, Alexandra; Aires, Tiago; Yergaliyev, Timur; Camarinha-Silva, Amélia; Vilanova, Manuel; Fonseca, António J. M.; Lima, Sofia A. C.; Maia, Margarida R. G.; Cabrita, Ana R. J.; Guilherme-Fernandes, Joana; REQUIMTE, LAQV, ICBAS, School of Medicine and Biomedical Sciences, University of Porto, Porto, Portugal; Barroso, Carolina; REQUIMTE, LAQV, ICBAS, School of Medicine and Biomedical Sciences, University of Porto, Porto, Portugal; Correia, Alexandra; i3S, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal; Aires, Tiago; SORGAL, Sociedade de Óleos e Rações S.A. S. João Ovar, Portugal; Yergaliyev, Timur; Hohenheim Center for Livestock Microbiome Research, HoLMiR, University of Hohenheim, Stuttgart, Germany; Camarinha-Silva, Amélia; Hohenheim Center for Livestock Microbiome Research, HoLMiR, University of Hohenheim, Stuttgart, Germany; Vilanova, Manuel; i3S, Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal; Fonseca, António J. M.; REQUIMTE, LAQV, ICBAS, School of Medicine and Biomedical Sciences, University of Porto, Porto, Portugal; Lima, Sofia A. C.; REQUIMTE, LAQV, ICBAS, School of Medicine and Biomedical Sciences, University of Porto, Porto, Portugal; Maia, Margarida R. G.; REQUIMTE, LAQV, ICBAS, School of Medicine and Biomedical Sciences, University of Porto, Porto, Portugal; Cabrita, Ana R. J.; REQUIMTE, LAQV, ICBAS, School of Medicine and Biomedical Sciences, University of Porto, Porto, Portugal
The inclusion of protein hydrolysates from animal by-products in petfood may enhance the industry’s sustainability, while contributing to dogs’ health due to their nutritional and functional properties. However, inconsistent results of protein hydrolysates on gut microbiota, immune responses, among other health parameters, have been observed in dogs. This study aimed to evaluate the effects of diets supplemented with 5% shrimp hydrolysate, compared to non-supplemented diets, on hematological parameters, serum chemistry profile, innate and adaptive immune function, and fecal microbiota composition in adult Beagle dogs during a 12-week feeding trial. Dietary inclusion of shrimp hydrolysate decreased blood eosinophils ( P = 0.017) and glucose levels ( P = 0.023), while increasing levels of white blood cells ( P = 0.002), platelets ( P = 0.038), neutrophils ( P = 0.036), and CD4 + ( P < 0.001) and CD8 + T cells ( P < 0.001) single producers of tumor necrosis factor-alpha. It also led to a more extensive proliferation of CD4 + T cells in response to LipL32 antigen ( P = 0.020) and to a decrease in superoxide production in stimulated cells ( P = 0.002). Additionally, supplemented diets increased Oscillospiraceae ( P-adj = 0.005) and Clostridia ( P-adj = 0.017) abundance, and decreased such of Sellimonas ( P-adj = 0.025). Overall, the results indicate that diet supplementation with shrimp hydrolysate at 5% modulates the immune response and fecal microbiota, highlighting its potential to be included in hypoallergenic and gastrointestinal diets, and in diets for diabetic dogs.
Fecal cortisol metabolites indicate increased stress levels in horses during breaking-in: a pilot study
(2025) Krieber, Julia; Nowak, Aurelia C.; Geissberger, Jakob; Illichmann, Oliver; Macho-Maschler, Sabine; Palme, Rupert; Dengler, Franziska; Krieber, Julia; Institute of Physiology and Pathophysiology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria, (A.C.N.);; Nowak, Aurelia C.; Institute of Physiology and Pathophysiology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria, (A.C.N.);; Geissberger, Jakob; Institute of Physiology and Pathophysiology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria, (A.C.N.);; Illichmann, Oliver; Institute of Physiology and Pathophysiology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria, (A.C.N.);; Macho-Maschler, Sabine; Experimental Endocrinology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria;; Palme, Rupert; Experimental Endocrinology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria;; Dengler, Franziska; Institute of Physiology and Pathophysiology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria, (A.C.N.);; Madigan, John
Sport horses are frequently exposed to situations that were identified as stressors, indicated by an increased cortisol release, which might impair animal welfare. However, while many studies deal with the impact of exercise, transport, and competition on stress in horses, little is known about the early phase of a horse’s sports career and studies investigating the stress level of young horses during breaking-in are limited. To compare stress levels in unridden horses, horses during breaking-in, and horses in training we collected fecal samples of young, unridden horses ( n = 28) and of horses in different training stages ( n = 13) and measured fecal cortisol metabolite (FCM) concentrations. Our preliminary results showed that FCM concentrations of unridden horses were significantly lower than those of horses in training (Mann–Whitney rank sum test, p < 0.001). Particularly in the first year under the saddle FCMs were significantly higher than in unridden horses (one way ANOVA + post hoc Holm–Sidak test, p < 0.05), with a tendency for FCM levels to decrease with time in training. Furthermore, we observed that within the group of ridden horses there was a larger range of variability in FCM levels, suggesting individual variations regarding their ability to deal with (training-induced) stress. These results indicate that breaking-in is a stressful time for young horses, underlining the importance of carrying out the initial training as carefully as possible.
The potential of gene editing and genomic selection in improving quantitative traits: A simulation study regarding dual-purpose chicken
(2024) Chuang, Edward; Bennewitz, Jörn
The practice of chick culling is the killing of day-old chicks of the layer line chicken. Because of their inability to lay eggs and inefficiency to grow meat, killing them instead of raising them is more economically efficient. In recent years, this practice has raised ethical concerns and led to the ban of this practice in several countries, including Germany and France. One of the several possible solutions is the use of dual-purpose chicken. Unlike the commercial layers, the dual-purpose chicken has abilities in both egg laying and meat production. Even though dual-purpose chicken has better meat growth performance compared to the commercial layers, the use of dual-purpose chicken is still challenging. One of the major challenges to improve dual-purpose chicken is the negative correlation between egg production and meat production. To cope with this negative correlation, some have proposed that gene editing could be a helpful tool to mitigate the problem. The aim of this simulation study is to explore the potential of gene editing to improve negatively correlated quantitative traits. To be more specific, given the aforementioned reasons, the negative correlation between the meat and egg production traits in chicken was chosen as the topic of this study. That is, chicken breeding programs that develop and improve dual-purpose chicken were simulated to discuss the potential of gene editing in animal breeding programs. Chapter 1 begins this thesis with a general introduction of the background to the problem of chick culling, the challenge of developing dual-purpose chicken, and the use of gene editing. Some previous simulation studies on the use of gene editing to improve quantitative traits in animal breeding programs is also introduced in this chapter. Chapter 2 presents a literature review that discusses how gene editing technologies could change the breeding and production of farm animals. In accordance with this thesis's aim, a specific focus was placed on improving quantitative traits. By reviewing the technical and social limitations of gene editing, this review provides a basis for further discussion of the feasibility of the gene-editing implementation as presented in the next chapters. Chapter 3 introduces the simulation framework to explore the potential of gene editing in chicken breeding programs. This framework first simulates two divergently selected lines that reflect the commercial layers and broilers. Based on these two lines, two alternative breeding programs were simulated. One is to develop a dual-purpose line from a layer line (the L-Pure Scenario), while the other selects and improves a synthetic line obtained from a cross between a layer line and a broiler line (the L-B cross scenario). In both breeding schemes, gene editing was integrated into the genomic selection scheme applying optimal contribution selection. The simulation presented in Chapter 4 considers multiplex genome editing that could edit five to one hundred single nucleotide variants at a time. These variants include the causal variants and other falsely edited variants. The results suggest that multiplex gene editing is beneficial in both the L-Pure and the L-B cross scenarios. It also demonstrated that when the risk of negative off-target effect exists, editing more nucleotides does not guarantee extra genetic gain. The simulation presented in Chapter 5 considers the introduction of major genes that would have larger effects on meat production. Here, only the L-Pure scenario is considered. Different to Chapter 4, in this chapter, editing is only possible with only one gene at a time. This chapter aims to study the effects of the numbers of major genes and the pleiotropy (on egg production and meat production). The results suggest that gene editing would be beneficial in dual-purpose chicken breeding programs when the gene is mildly pleiotropic. Chapter 6 ends the thesis with a general discussion that covers the limitations of this study and the possibilities to further expand the simulation study.
An evaluation of the lineage of Brucella isolates in turkey by a whole-genome single-nucleotide polymorphism analysis
(2024) Akar, Kadir; Holzer, Katharina; Hoelzle, Ludwig E.; Yıldız Öz, Gülseren; Abdelmegid, Shaimaa; Baklan, Emin Ayhan; Eroğlu, Buket; Atıl, Eray; Moustafa, Shawky A.; Wareth, Gamal; Elkhayat, Manar; Akar, Kadir; Faculty of Veterinary Medicine, Van Yuzuncu Yıl University, 65090 Van, Turkey; Holzer, Katharina; Department of Livestock Infectiology and Environmental Hygiene, Institute of Animal Science, University of Hohenheim, 70599 Stuttgart, Germany;; Hoelzle, Ludwig E.; Department of Livestock Infectiology and Environmental Hygiene, Institute of Animal Science, University of Hohenheim, 70599 Stuttgart, Germany;; Yıldız Öz, Gülseren; NRL for Brucellosis, Pendik Veterinary Control Institute, 34890 Istanbul, Turkey; (G.Y.Ö.); (E.A.B.); (B.E.); (E.A.); Abdelmegid, Shaimaa; Faculty of Veterinary Medicine, Assiut University, Assiut 71515, Egypt;; Baklan, Emin Ayhan; NRL for Brucellosis, Pendik Veterinary Control Institute, 34890 Istanbul, Turkey; (G.Y.Ö.); (E.A.B.); (B.E.); (E.A.); Eroğlu, Buket; NRL for Brucellosis, Pendik Veterinary Control Institute, 34890 Istanbul, Turkey; (G.Y.Ö.); (E.A.B.); (B.E.); (E.A.); Atıl, Eray; NRL for Brucellosis, Pendik Veterinary Control Institute, 34890 Istanbul, Turkey; (G.Y.Ö.); (E.A.B.); (B.E.); (E.A.); Moustafa, Shawky A.; Faculty of Veterinary Medicine, Benha University, Toukh 13736, Egypt; (S.A.M.); (M.E.); Wareth, Gamal; Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institute, 07743 Jena, Germany;; Elkhayat, Manar; Faculty of Veterinary Medicine, Benha University, Toukh 13736, Egypt; (S.A.M.); (M.E.); Pedersen, Karl
Brucellosis is a disease caused by the Brucella ( B. ) species. It is a zoonotic disease that affects farm animals and causes economic losses in many countries worldwide. Brucella has the ability to persist in the environment and infect the host at low doses. Thus, it is more important to trace brucellosis outbreaks, identify their sources of infection, and interrupt their transmission. Some countries already have initial data, but most of these data are based on a Multiple-Locus Variable-Number Tandem-Repeat Analysis (MLVA), which is completely unsuitable for studying the Brucella genome. Since brucellosis is an endemic disease in Turkey, this study aimed to examine the genome of Turkish Brucella isolates collected between 2018 and 2020, except for one isolate, which was from 2012. A total of 28 strains of B. melitensis ( n = 15) and B. abortus ( n = 13) were analyzed using a core-genome single-nucleotide polymorphism (cgSNP) analysis. A potential connection between the Turkish isolates and entries from Sweden, Israel, Syria, Austria, and India for B. melitensis was detected. For B. abortus , there may be potential associations with entries from China. This explains the tight ties found between Brucella strains from neighboring countries and isolates from Turkey. Therefore, it is recommended that strict measures be taken and the possible effects of uncontrolled animal introduction are emphasized.
Subconjunctival bleedings in neonatal calves: a case series report
(2022) Steffl, Martin; Nautscher, Nadine; Steffl, Martin; Faculty of Agricultural Sciences, Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Nautscher, Nadine; Faculty of Agricultural Sciences, Institute of Animal Science, University of Hohenheim, Stuttgart, Germany
Background: In animals, only few reports exist about the occurrence and causes of subconjunctival bleedings, especially in newborn calves. Most case reports and studies showed that the major risk factors for subconjunctival bleedings in animals are traumatic events such as birth trauma and traffic accidents, respectively. In neonatal babies, it is suggested that compression of the thorax and abdomen during delivery or forces generated in utero during labor may raise venous pressure to conjunctival vessels and can cause subconjunctival bleedings. Results: The incidence of bleedings in neonatal Holstein–Friesian calves was 2.4 per cent of 289 neonatal calves examined over a six-year period. In general, two types of subconjunctival bleedings were seen. One was usually in a semilunar fashion immediately outside the limbus of the eye. The other type was a stripe or macule of variable size at different positions of the sclera. The subconjunctival bleedings were not related to gestational time. In all cases, affected calves were born without assistance. Multiparous cows were more often involved in the calves with subconjunctival bleedings. Two calves examined haematologically did not show signs of anemia or thrombocytopenia. Conclusions: Subconjunctival bleedings in neonatal calves appear not to be incidental findings. Main causes or associated conditions of subconjunctival bleedings were not found.
Endogenous mucosal phosphatases characterization in duodenum brush border membrane of laying hens
(2025) Hanauska, Anna; Sommerfeld, Vera; Schollenberger, Margit; Huber, Korinna; Rodehutscord, Markus
Chicken mucosal phosphatases can partially degrade phytate contained in the feed. Little is known about the characteristics and degradation products of such mucosal phosphatases and the effects of age and genetic strain of the chicken. The objective of this study was to characterize endogenous mucosal phosphatases of two laying hen strains fed diets with or without mineral phosphorus (P) before and after the onset of egg laying. Hens of the strains Lohmann Brown-classic (LB) and Lohmann LSL-classic (LSL) were sacrificed in weeks 19 and 24 of age after 4 weeks of feeding one of two diets with (P+) or without (P-) mineral P supplement. Mucosa of the duodenum was collected, and the brush border membrane (BBM) of enterocytes was enriched and used for phosphatase activity determination. Additionally, the BBM was used in a modified three-step in vitro assay to study the InsP6 degradation products. The results of both in vitro assays were not significantly affected by hen strain and diet. The activity of mucosal phosphatase in 19-week-old hens was, on average, 0.8 µmol Pi/g BBM protein/min lower than in 24-week-old hens (P < 0.002). Consistently, the InsP6 concentration in the incubation residue was significantly higher in 19-week-old hens than in 24-week-old hens (P < 0.001). In the incubation residue, the concentrations of Ins(1,2,3,4,5)P5, Ins(1,2,3,4,6)P5, and Ins(1,2,3,4)P4 were significantly lower (P ≤ 0.002), and those of InsP3 and InsP2 were significantly higher (P ≤ 0.027) when BBM of 24-week-old hens was used compared to 19-week-old hens. The InsP6 degradation products suggest the activity primarily of a 6- and secondarily of a 5-phytase in the duodenal mucosa. The consistent results from both in vitro assays provide a comprehensive characterization of these enzymes. Under the conditions of this study, small intestine calcium concentration appeared to influence mucosal enzyme activity more than dietary mineral P supplementation.
Composition of the ileum microbiota is a mediator between the host genome and phosphorus utilization and other efficiency traits in Japanese quail (Coturnix japonica)
(2022) Haas, Valentin; Vollmar, Solveig; Preuß, Siegfried; Rodehutscord, Markus; Camarinha-Silva, Amélia; Bennewitz, Jörn; Haas, Valentin; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Vollmar, Solveig; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Preuß, Siegfried; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Rodehutscord, Markus; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Camarinha-Silva, Amélia; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Bennewitz, Jörn; Institute of Animal Science, University of Hohenheim, Stuttgart, Germany
Background: Phosphorus is an essential nutrient in all living organisms and, currently, it is the focus of much attention due to its global scarcity, the environmental impact of phosphorus from excreta, and its low digestibility due to its storage in the form of phytates in plants. In poultry, phosphorus utilization is influenced by composition of the ileum microbiota and host genetics. In our study, we analyzed the impact of host genetics on composition of the ileum microbiota and the relationship of the relative abundance of ileal bacterial genera with phosphorus utilization and related quantitative traits in Japanese quail. An F2 cross of 758 quails was genotyped with 4k genome-wide single nucleotide polymorphisms (SNPs) and composition of the ileum microbiota was characterized using target amplicon sequencing. Heritabilities of the relative abundance of bacterial genera were estimated and quantitative trait locus (QTL) linkage mapping for the host was conducted for the heritable genera. Phenotypic and genetic correlations and recursive relationships between bacterial genera and quantitative traits were estimated using structural equation models. A genomic best linear unbiased prediction (GBLUP) and microbial (M)BLUP hologenomic selection approach was applied to assess the feasibility of breeding for improved phosphorus utilization based on the host genome and the heritable part of composition of the ileum microbiota. Results: Among the 59 bacterial genera examined, 24 showed a significant heritability (nominal p ≤ 0.05), ranging from 0.04 to 0.17. For these genera, six genome-wide significant QTL were mapped. Significant recursive effects were found, which support the indirect host genetic effects on the host’s quantitative traits via microbiota composition in the ileum of quail. Cross-validated microbial and genomic prediction accuracies confirmed the strong impact of microbial composition and host genetics on the host’s quantitative traits, as the GBLUP accuracies based on the heritable microbiota-mediated components of the traits were similar to the accuracies of conventional GBLUP based on genome-wide SNPs. Conclusions: Our results revealed a significant effect of host genetics on composition of the ileal microbiota and confirmed that host genetics and composition of the ileum microbiota have an impact on the host’s quantitative traits. This offers the possibility to breed for improved phosphorus utilization based on the host genome and the heritable part of composition of the ileum microbiota.
Occurrence of ‘Candidatus Mycoplasma haemosuis’ in fattening pigs, sows and piglets in Germany using a novel gap-based quantitative real-time PCR assay
(2022) Ade, Julia; Stadler, Julia; Ritzmann, Mathias; Zübert, Christina; Hoelzle, Katharina; Hoelzle, Ludwig E.; Ade, Julia; Institute of Animal Science, Department of Livestock Infectiology and Environmental Hygiene, University of Hohenheim, Stuttgart, Germany; Stadler, Julia; Clinic for Swine, Centre for Clinical Veterinary Medicine, Ludwig-Maximilians-Universität Munich, Oberschleissheim, Germany; Ritzmann, Mathias; Clinic for Swine, Centre for Clinical Veterinary Medicine, Ludwig-Maximilians-Universität Munich, Oberschleissheim, Germany; Zübert, Christina; Institute of Animal Science, Department of Livestock Infectiology and Environmental Hygiene, University of Hohenheim, Stuttgart, Germany; Hoelzle, Katharina; Institute of Animal Science, Department of Livestock Infectiology and Environmental Hygiene, University of Hohenheim, Stuttgart, Germany; Hoelzle, Ludwig E.; Institute of Animal Science, Department of Livestock Infectiology and Environmental Hygiene, University of Hohenheim, Stuttgart, Germany
Background: The appearance of the novel porcine haemotrophic mycoplasma (HM) species ‘Candidatus Mycoplasma haemosuis’ was reported in apparently healthy but also in clinically sick animals in China, Korea and in a case report from Germany. Outside of Asia, however, nothing further is known about the frequency of ‘Ca. M. haemosuis’ in pigs to date. To investigate the distribution of this novel HM species in Germany, fattening pigs, sows and pre-suckling piglets were examined using a herein developed quantitative real-time PCR assay (qPCR). Because the piglets were sampled before the first colostrum uptake, additional information on a possible vertical transmission from dams to their offspring was obtained. Results: Our novel qPCR assay successfully detected ‘Ca. M. haemosuis’ in all blood samples from the ‘Ca. M. haemosuis’-infected pigs. No cross-reactivity was detected when DNA from non-target Mycoplasma spp. and other bacterial species representing 105 bacteria/reaction were used as a template. The lower limit of detection of the qPCR was thus 10 gap gene copies per reaction and 2.5 x 103 genome equivalents (GE) per mL blood. ‘Candidatus M. haemosuis’ was detected by this qPCR in blood samples from a total out of 6.25% sows (13/208), 4.50% pre-suckling piglets (28/622) and 17.50% fattening pigs (35/200). On farm level, 3 out of 21 piglet producing farms (14.28%) and 9 out of 20 fattening farms (45.00%) were positive for ‘Ca. M. haemosuis’. Co-infections with M. suis were evident in all age groups. Conclusion: ‘Candidatus M. haemosuis’ infection is present in German pig farms and the detection of the novel porcine HM species in piglets immediately after birth before colostrum intake indicates vertical transmission. The novel qPCR assay specific for ‘Ca. M. haemosuis’ described herein will be a prerequisite for future studies on the prevalence, epidemiology as well as the clinical and economic impact of ‘Ca. M. haemosuis’ infections.
Structural variants and tandem repeats in the founder individuals of four F2 pig crosses and implications to F2 GWAS results
(2022) Blaj, Iulia; Tetens, Jens; Bennewitz, Jörn; Thaller, Georg; Falker-Gieske, Clemens; Blaj, Iulia; Institute of Animal Breeding and Husbandry, Kiel University, Kiel, Germany; Tetens, Jens; Center for Integrated Breeding Research, Georg-August-University, Göttingen, Germany; Bennewitz, Jörn; Institute of Animal Husbandry and Breeding, University of Hohenheim, Stuttgart, Germany; Thaller, Georg; Institute of Animal Breeding and Husbandry, Kiel University, Kiel, Germany; Falker-Gieske, Clemens; Department of Animal Sciences, Georg-August-University, Göttingen, Germany
Background: Structural variants and tandem repeats are relevant sources of genomic variation that are not routinely analyzed in genome wide association studies mainly due to challenging identification and genotyping. Here, we profiled these variants via state-of-the-art strategies in the founder animals of four F2 pig crosses using whole-genome sequence data (20x coverage). The variants were compared at a founder level with the commonly screened SNPs and small indels. At the F2 level, we carried out an association study using imputed structural variants and tandem repeats with four growth and carcass traits followed by a comparison with a previously conducted SNPs and small indels based association study. Results: A total of 13,201 high confidence structural variants and 103,730 polymorphic tandem repeats (with a repeat length of 2-20 bp) were profiled in the founders. We observed a moderate to high (r from 0.48 to 0.57) level of co-localization between SNPs or small indels and structural variants or tandem repeats. In the association step 56.56% of the significant variants were not in high LD with significantly associated SNPs and small indels identified for the same traits in the earlier study and thus presumably not tagged in case of a standard association study. For the four growth and carcass traits investigated, many of the already proposed candidate genes in our previous studies were confirmed and additional ones were identified. Interestingly, a common pattern on how structural variants or tandem repeats regulate the phenotypic traits emerged. Many of the significant variants were embedded or nearby long non-coding RNAs drawing attention to their functional importance. Through which specific mechanisms the identified long non-coding RNAs and their associated structural variants or tandem repeats contribute to quantitative trait variation will need further investigation. Conclusions: The current study provides insights into the characteristics of structural variants and tandem repeats and their role in association studies. A systematic incorporation of these variants into genome wide association studies is advised. While not of immediate interest for genomic prediction purposes, this will be particularly beneficial for elucidating biological mechanisms driving the complex trait variation.
Determination of optimal phage load and administration time for antibacterial treatment
(2024) Plunder, Steffen; Burkard, Markus; Helling, Thomas; Lauer, Ulrich M.; Hoelzle, Ludwig E.; Marongiu, Luigi
Using phages as antibacterials is becoming a customary practice in Western countries. Nonetheless, successful treatments must consider the growth rate of the bacterial host and the degradation of the virions. Therefore, successful treatments require administering the right amount of phage (viral load, Vφ) at the right moment (administration time, Tφ). The present protocols implement a machine learning approach to determine the best combination of Vφ and Tφ to obtain the elimination of the target bacterium from a system. Basic Protocol 1: One bacterium, one phage. Alternate Protocol 1: One bacterium, one phage (wrapping function). Alternate Protocol 2: One bacterium, one phage (wrapping function, alternative growing model). Basic Protocol 2: Two bacteria, one phage. Alternate Protocol 3: Two bacteria, one phage (launch from terminal).

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