Institut für Nutztierwissenschaften

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Parasites, depredators, and limited resources as potential drivers of winter mortality of feral honeybee colonies in German forests
(2023) Kohl, Patrick L.; Rutschmann, Benjamin; Sikora, Luis G.; Wimmer, Norbert; Zahner, Volker; D’Alvise, Paul; Hasselmann, Martin; Steffan-Dewenter, Ingolf; Kohl, Patrick L.; Department of Animal Ecology and Tropical Biology, Biocenter, University of Würzburg, Würzburg, Germany; Rutschmann, Benjamin; Department of Animal Ecology and Tropical Biology, Biocenter, University of Würzburg, Würzburg, Germany; Sikora, Luis G.; NaturKonzept, Pfullingen, Germany; Wimmer, Norbert; Bayerische Landesanstalt Für Wald Und Forstwirtschaft, Freising, Germany; Zahner, Volker; Forest Ecology and Management, University of Applied Sciences Weihenstephan-Triesdorf, Freising, Germany; D’Alvise, Paul; Institute for Medical Microbiology and Hygiene, University Hospital Tübingen, Tübingen, Germany; Hasselmann, Martin; Department of Livestock Population Genomics, Institute of Animal Science, University of Hohenheim, Stuttgart, Germany; Steffan-Dewenter, Ingolf; Department of Animal Ecology and Tropical Biology, Biocenter, University of Würzburg, Würzburg, Germany
Wild honeybees ( Apis mellifera ) are considered extinct in most parts of Europe. The likely causes of their decline include increased parasite burden, lack of high-quality nesting sites and associated depredation pressure, and food scarcity. In Germany, feral honeybees still colonize managed forests, but their survival rate is too low to maintain viable populations. Based on colony observations collected during a monitoring study, data on parasite prevalence, experiments on nest depredation, and analyses of land cover maps, we explored whether parasite pressure, depredation or expected landscape-level food availability explain feral colony winter mortality. Considering the colony-level occurrence of 18 microparasites in the previous summer, colonies that died did not have a higher parasite burden than colonies that survived. Camera traps installed at cavity trees revealed that four woodpecker species, great tits, and pine martens act as nest depredators. In a depredator exclusion experiment, the winter survival rate of colonies in cavities with protected entrances was 50% higher than that of colonies with unmanipulated entrances. Landscapes surrounding surviving colonies contained on average 6.4 percentage points more cropland than landscapes surrounding dying colonies, with cropland being known to disproportionately provide forage for bees in our study system. We conclude that the lack of spacious but well-protected nesting cavities and the shortage of food are currently more important than parasites in limiting populations of wild-living honeybees in German forests. Increasing the density and diversity of large tree cavities and promoting bee forage plants in forests will probably promote wild-living honeybees despite parasite pressure.
Approaches to increase digestibility of Bangladesh ruminant feed resources in order to mitigate enteric methane production
(2024) Bashar, Muhammad Khairul; Rodehutscord, Markus
Multiple analyses have shown that the rising human population, urbanization, and consumer preferences affect the demand for livestock products in developing countries. Concurrently, human population and urbanisation growth are reducing the likelihood of newly cultivated land producing feeds or restoring damaged pastures. Using low-quality roughage more efficiently is an option, but such roughages have drawbacks, including high structural carbohydrate and low nitrogen contents, which lead to poor palatability and nutrient utilisation including emission of methane (CH4) that is harmful to the environment and a loss of energy to the animal. Using commonly accessible legumes and tree foliage could be a possibility to address the need for N and CH4 mitigation due to their high crude protein and secondary compound contents. In order to evaluate the potential of specific combinations of roughages, concentrates, and other feeds, it is necessary to determine the feeding values, in vitro ruminal fermentation, and CH4 production of single feeds, as well as the promising interaction (associative) effects between single feeds when combinded in a ration. This may allow for formulating a better balanced total mixed ration (TMR) for ruminants, which was the main goal of the present doctoral study. Eighteen feedstuffs from Bangladesh were chosen, including roughages such as crop residues (rice straw, urea molasses treated straw and maize stover), silages (Napier silage and Maize silage), common grasses (German grass, Para grass and Napier grass) and leguminous fodder (Ipil-ipil, Glicidia, Alfalfa hay and Moringa tops) and concentrates (crushed wheat, crushed maize, Wheat bran, Kashari bran, and Rice bran). In Study 1, the single feeds were characterized by comprehensive chemical analysis and in vitro production of total gas and methane using the Hohenheim gas test. Based on the results of Study 1, rice straw, German grass, Napier silage, and maize silage were used in Study 2 and incubated in vitro alone or after substitution by Ipil-ipil and Gliricidia at levels of 10%, 20%, 30%, and 40%, respectively. In Study 3, mixtures of roughages and leguminous fodder (rice straw + 40% Ipil-ipil, German grass + 10% Ipil-ipil, Napier silage + 30% Ipil ipil, and maize silage + 30% Ipil-ipil) were combined with single concentrates in order to identify the most promising mixtures in regard to digestibility and methane production. Eventually, 24 TMR were formulated using six for each type of roughage in Study 4. These TMR were comprised of 60% of the same combination of roughage as used in Study 3 and 40% of various combinations of concentrates including TMR-1 (60% rice bran + 10% wheat bran + 30% kashari bran), TMR-2 (30% rice bran + 10% wheat bran + 60% kashari bran), TMR-3 (10% rice bran + 60% wheat bran + 30% kashari bran), TMR-4 (10% rice bran + 30% wheat bran + 60% kashari bran), TMR-5 (60% rice bran + 30% wheat bran + 10 kashari bran), and TMR-6 (30% rice bran + 60% wheat bran + 10% kashari bran) in order to identify suitable TMR for maximising ruminal fermentation and reducing the CH4 production in vitro. In all studies, the total gas production (GP), digestibility of organic matter (dOM), metabolisable energy (ME), and net energy for lactation (NEL) were determined after incubating 200 mg of feed with a rumen fluid-buffer solution for 72 hours. In addition, the CH4 concentration in the produced gas was measured after incubating 120 mg of feed for 24 hours. Measured values of the mixed feed were compared to estimated values, where the estimated values were weighted means of the values of the individual feeds, to determine the associative effects between the feed ingredients (Study 2 and Study 4). In Study 1, the concentration of acid detergent fibre (ADF), neutral detergent fibre (NDF), and acid detergent lignin (ADL) was the lowest in leguminous fodder while the crude protein (CP) concentration was the hightest, followed by the common grasses and the silages. The crop residues showed the lowest CP and the highest cell wall fraction concentrations. The dOM, ME, and NEL of crushed wheat and crushed maize were significantly higher (p<0.05) than those of other feedstuffs. The same differences were found (p<0.05) for CH4 concentration (% of GP) and CH4 production (L CH4/kg dOM). The dOM and ME of German grass and Ipil-ipil were higher (p<0.05), whereas the CH4 concentration and CH4 production were lower compared to that of crop residues and other common grasses. The CH4 production of single feeds decreased with increasing concentrations of CP, ADF, and ADL, whereas it increased with NDF concentration. These findings enabled development of more balanced diets for ruminants with the aim of improved digestibility and reduced CH4 emission while making use of widely available feed resources in Bangladesh. Study 2 aimed to evaluate the in vitro ruminal fermentation, CH4 production, and associative effects between low-quality roughages and CP supplements. The gas production after 24 hours (GP24) and rumen fermentation rate increased (p<0.05) with increasing addition of Ipil-ipil to rice straw. It also resulted in a gradual increase of dOM and ME (p<0.05) of the mixture, reaching maximum values (59.1% and 7.60 MJ ME/kg DM) and maximum reductions in CH4 concentration and CH4/dOM (6.9% and 11.7%) compared to the control with Ipil-ipil addition at 40% of DM. When Ipil-ipil was added at 30% to Napier silage, the same trend was observed for dOM and ME (56.5%, and 7.55 MJ ME/kg DM) and CH4 concentration and CH4/dOM (5.3% and 13.3%). Ipil-ipil did not significantly interact with high quality roughages such as German grass and maize silage. Increasing the addition of Gliricidia to the roughages in Study 2 led to a decrease in GP24, dOM, ME, and CH4 production. The highest levels of the associative effects (p<0.05) were seen when 40% Ipil-ipil was added to rice straw and 30% was added to Napier silage. In conclusion, adding Ipil-ipil to low-quality roughages is superior to Gliricidia and showed promising results, with the ranking as follows: rice straw + 40% Ipil-ipil > Napier silage + 30% Ipil-ipil > maize silage + 30% Ipil-ipil > German gras + 10% Ipil-ipil. Study 3 and Study 4 had the overarching purpose to determine which combination of single concentrates and combinations of concentrates with roughage produced the best mixed feed in terms of rumen fermentation, CH4 reduction potential, and associative effect. The CP and non-stach carbohydrate contents of the Ipil-ipil-based mixed feed was enhanced by adding Wheat bran and Kashari bran as single concentrates, which had a favourable effect on rumen fermentation and gas production but had the opposite effect on CH4 production when compared to the addition of Rice bran. In Study 4, TMR were formulated using roughages and Ipil-ipil and addition of Wheat bran, Kashari bran, and Rice bran in various combinations. The TMR based on German grass and maize silage with specific bran combinations showed significant reductions in in vitro CH4 production, CH4 concentration, and CH4/dOM while maximising GP, dOM, ME, and the associative effects. To conclude, the results of chemical analyses and in vitro fermentation studies showed that specific combinations of roughages, protein feeds, and by-products available in Bangladesh have the potential to formulate rations for cattle that help making livestock production more sustainable. The results may be also relevant for other developing nations. It is suggested to verify the results of the present project by animal trials at local conditions.
Review: Welfare in farm animals from an animal-centred point of view
(2024) Huber, Korinna
This review aimed to enlighten aspects of welfare from the farm animal-centred point of view rarely addressed such as those anatomical and physiological alterations induced in farm animals to obtain high performance. Hence, the major working hypothesis was that high-producing farm animals developed an imbalance between body structural and functional capacities and the genetic procedures applied to obtain industrial production of animal protein. This is called “disproportionality”, a feature which cannot be compensated by feeding and management approaches. Consequences of disproportionality are the insidious development of disturbances of the metabolism, low-grade systemic inflammation and as a final stage, production diseases, developing throughout the productive life span of a farm animal and affecting animal welfare. Based on scientific evidence from literature, the review discusses disproportional conditions in broilers, laying hens, sows, piglets, dairy cows, bulls and calves as the most important farm animals for production of milk, meat, foetuses and eggs. As a conclusion, farm animal welfare must consider analysing issues from an animal-centered point of view because it seems evident that, due to genetics and management pressures, most of farm animals are already beyond their physiological limitations. Animal welfare from an animal-centered point must be addressed as an ethical step to establish limits to the strength placed on the animal’s anatomical and physiological functionality. It may allow more sustainable and efficient farm animal production and the availability of healthy animal-derived protein for human nutrition worldwide.
Studies of phytate degradation and associated mineral utilization in growing pigs
(2024) Klein, Nicolas; Rodehutscord, Markus
Phosphorus (P) is primarily stored in plant seeds as phytate, which is any salt of phytic acid (myo-inositol hexakisphosphate; InsP6). The enzymatic capacity of pigs and the microorganisms present in their digestive tract allows for only partial and incomplete prececal degradation of InsP6. To address this limitation, the addition of exogenous phytases to feed has been state-of-the-art for more than three decades. Phytases, found in various organisms like bacteria, fungi, plants, and animals, initiate the breakdown of InsP6 and inorganic phosphate, thereby making the liberated phosphate available for digestion. Substantial scientific research and practical field applications have shown that adding exogenous phytases to the diet of non-ruminants leads to an improvement in P digestibility and a reduction in P excretion. Over time, phytases with enhanced thermal profiles and higher activity at physiological pH milieus have been engineered. Additionally, advancements in plant breeding have increased intrinsic phytase activity in feed ingredients relevant to animal nutrition. Thus, this thesis aimed to deepen the understanding of inositol phosphate (InsP) degradation in ileal-cannulated pigs, focusing on how dietary variables can influence this process along the digestive tract. In the first study (Manuscript 1), diets based on wheat with variable intrinsic phytase activity and the addition of an exogenous phytase on InsP degradation in pigs were investigated. Although wheat intrinsic phytase contributed to increased InsP degradation and P digestibility in pigs, variation in intrinsic phytase activity by crossbreeding wheat, as achieved in this study, was not reflected in InsP degradation and P digestibility data. This discrepancy may be attributed to an inadequate proteolytic resistance of wheat phytase in the gastric compartment of the pig. A marked increase in prececal InsP degradation and P digestibility was achieved by adding a microbial phytase to the wheat-based feed. While exogenous phytase increases the digestibility of InsP-P, other dietary components may impair its hydrolytic activity. In the second study (Manuscript 2), the single and interactive effects of dietary Ca concentration and exogenous phytase in pigs were investigated. The study assessed the effects of dietary Ca and exogenous phytase on InsP degradation and nutrient digestibility in growing pigs. Additional dietary Ca decreased prececal InsP6 disappearance, but only with exogenous phytase. Concentrations of lower InsP isomers and myo-inositol in the ileal digesta and prececal P digestibility were greater with exogenous phytase, but not affected by dietary Ca concentration. In contrast, fecal InsP6 disappearance was lower and the concentration of InsP4-5 isomers in feces was greater with additional dietary Ca. The study revealed that dietary Ca concentration is relevant for InsP disappearance in the hindgut, but not in the ileum. However, when exogenous phytase is used, the dietary Ca concentration is important because prececal InsP degradation is affected by the Ca level in the diet. The third study (Manuscript 3) aimed to investigate the effects of dietary Ca sources, exogenous phytase, and formic acid on InsP degradation and nutrient digestibility in growing pigs. Replacing limestone with Ca formate reduced prececal InsP6 disappearance and prececal P digestibility. However, adding formic acid to a diet containing limestone and phytase enhanced prececal InsP6 disappearance and increased InsP2 and myo-inositol concentrations in the distal ileal digesta. Accordingly, prececal P digestibility was increased when formic acid was added to the diet containing limestone and exogenous phytase. This suggested that Ca formate is not a suitable alternative Ca source to limestone for optimizing prececal P release from InsP in growing pigs. The release of P from InsP by exogenous phytase in limestone diets can still be further increased by adding formic acid. In conclusion, the degradation of InsP in growing pigs is a complex process affected by dietary Ca concentration, dietary Ca source, and the presence of phytase. However, investigated dietary traits mainly affected InsP6, with lesser effects on lower InsP isomers, highlighting the importance of prececal hydrolytic breakdown of InsP6 to InsP5. This directly affected P digestibility of growing pigs. Intrinsic phytase activity in feed ingredients, such as wheat, can contribute to prececal InsP degradation but may be neglected when exogenous phytase is present in the feed. Exogenous phytase addition notably increased prececal InsP degradation and P digestibility in plant-based diets, and complete dephosphorylation of InsP in the prececal tract was implied by the increase of the final degradation product myo-inositol in the ileum content, urine, and blood. However, some P remained bound to InsP at the end of the ileum of growing pigs and was therefore not available for digestion. Nevertheless, some diets with exogenous phytase tested in this thesis met the recommended digestible P concentration. This suggests that diets formulated with adequate InsP-containing ingredients and exogenous phytase may obviate the need for mineral P addition, reducing the reliance on finite P sources in growing pig nutrition.
Factors influencing proteolysis and protein utilization in the intestine of pigs: A review
(2021) Kurz, Alina; Seifert, Jana
Pigs are among the most important farm animals for meat production worldwide. In order to meet the amino acid requirements of the animals, pigs rely on the regular intake of proteins and amino acids with their feed. Unfortunately, pigs excrete about two thirds of the used protein, and production of pork is currently associated with a high emission of nitrogen compounds resulting in negative impacts on the environment. Thus, improving protein efficiency in pigs is a central aim to decrease the usage of protein carriers in feed and to lower nitrogen emissions. This is necessary as the supply of plant protein sources is limited by the yield and the cultivable acreage for protein plants. Strategies to increase protein efficiency that go beyond the known feeding options have to be investigated considering the characteristics of the individual animals. This requires a deep understanding of the intestinal processes including enzymatic activities, capacities of amino acid transporters and the microbiome. This review provides an overview of these physiological factors and the respective analyses methods.
Standard Methods for Dissection of Varroa destructor Females
(2021) Piou, Vincent; Vilarem, Caroline; Rein, Carolin; Sprau, Lina; Vétillard, Angélique
Varroa destructor (Anderson and Trueman) is known as a major pest of Apis mellifera L, especially in the Northern Hemisphere where its effects can be deleterious. As an obligate parasite, this mite relies entirely on its host to reproduce and complete its cycle. Studies focusing on isolated organs are needed to better comprehend this organism. To conduct such targeted molecular or physiological studies, the dissection of V. destructor mites is crucial as it allows the extraction of specific organs. Here, we propose a technical article showing detailed steps of females V. destructor dissection, illustrated with pictures and videos. These illustrated guidelines will represent a helpful tool to go further in V. destructor research.
Nutrient utilisation and metabolic adaptation of broiler chickens to high levels of free amino acids in the diet
(2024) Ibrahim, Ahmad; Rodehutscord, Markus
Reducing dietary crude protein (CP) content in broiler chickens is a strategy used to reduce nitrogen excretion and its impact on the environment caused by the broiler industry. This requires an increased proportion of free amino acids (AA) in the diet to avoid insufficient AA supply. An upper limit of dietary free AA inclusion has often been suggested but could not be derived from the current literature. The overall objective of this dissertation was to determine the upper limit of free AA content in CP-reduced diets. Additionally, the aim was to investigate the effects of adding free AA on the acid-base status in the blood of broiler chickens. Furthermore, the physiological and metabolic responses to substituting peptide-bound AA with free AA were investigated. In a preliminary experiment of the first study, the precaecal AA digestibility of soya protein isolate (SPI) representing a peptide-bound AA source was determined. One diet with 80 g SPI per kg and another with the basal mix, including maize starch in substitution with SPI, were tested. The diets were offered from day 18–21 of age and tested in eight units with 15 animals each. On day 21, the birds were slaughtered and the content of the last distal ileum was sampled. The amount of precaecal digestible AA derived from SPI was calculated as the difference in AA digestibility between both diets. In the main trial of the first study, the amount of digestible AA derived from SPI was incrementally substituted with a free AA mixture, i.e. 0, 25, 50, 75, and 100 % to determine the maximum amount of peptide-bound AA that can be substituted by free AA without reducing growth performance. At each substitution level, the amounts of digestible aspartate (Asp) and glutamate (Glu), which, per analysis, also contained asparagine (Asn) and glutamine (Gln), were either substituted with free Asp and Glu or with a 50/50 mixture of Asp/Asn and Glu/Gln. Each diet was offered from day 7–21 of age and tested in 7 units of 10 animals each. Based on growth performance, there was a maximum limit of dietary free AA inclusion dependent on the inclusion of Asn and Gln. This indicates that Asn and/or Gln were limited in the diets including only Asp and Glu and high levels of free AA. Asn and Gln supply allowed for increased AA substitution from 10 to 19 % of CP in the diet and increased free AA concentration from about 37 to 54 g/kg without reducing growth. Additionally, as the AA substitution was done in 25 %-unit increments, the maximum amount of dietary free AA was estimated between 54–71 g/kg. The broken line linear regression estimated value of 57 g free AA/kg in the diets with Asn and Gln without impairing growth and 61 g free AA/kg without impairing feed intake. Blood data related to the acid-base balance indicated a compensated acidosis 14 days after the diet change, which was attenuated by including Asn and Gln in the diet. In the second study, three diets were selected based on the results of the first study. The first diet included SPI with 0 % AA substitution. The second diet included the maximum free AA level with 50 % AA substitution without negative effects on growth and nitrogen utilisation. The third diet with 100 % AA substitution had evidence of an acid-base shift. The second and third diets considered all 20 proteinogenic AA including Asn and Gln. The change to one of these three diets occurred on day 7 post-hatch and feeding continued until day 22. Excreta were collected on days 7–8, 8–9, 9–10, 11–12, 14–15, and 21–22. Repeated blood samples were withdrawn on days 7, 8, 9, 11, 14, and 21. There was no evidence of affected nitrogen utilisation by substituting peptide-bound with free AA, especially 3 days after the diet change. It was confirmed that up to 50 % AA substitution from SPI did not affect growth. The reduction in growth at 100 % AA substitution was mainly due to the reduced feed intake immediately after the diet change. Adaption of nitrogen utilisation and AA in blood plasma to high levels of dietary free AA occurred within 3 to 7 days after changing to 50 % and 100 % AA substitution diets, respectively. This was suggested by the data on feed intake, nitrogen accretion, nitrogen utilisation efficiency in the first 3 days and plasma free AA concentrations in the first 7 days after the diet change, which remained unaffected by the treatments thereafter. A significant increase in ammonia excretion was also observed with increasing levels of free AA at all points in time. The blood samples were also investigated for responses in the acid-base balance and plasma metabolites directly after the diet change. An acid-base shift was determined in the birds that received 100 % and 50 % AA substitution diets. An acute reaction of the acid-base balance was on days 4 to 7 and day 4 after a change to the 100 % and 50 % AA substitution diets, respectively. Thereafter, a compensated acid-base acidosis was determined on day 7 for 50 % AA substitution and on day 14 for 100 % AA substitution. This likely explained the increased ammonia excretion found for these diets as an adaptation to excrete acids after an acid-base shift. The untargeted metabolomics analysis effectively determined the reduction in the use of plant feed ingredients with increasing free AA inclusion. However, no changes in metabolic pathways by increasing free AA inclusion were detected. It can be concluded that there was a maximum limit to including free AA in diets for broiler chickens, which was dependent on the supply of Asn and Gln. A reduced feed intake was the primary response of the birds to this upper limit. One of the likely reasons for the reduced feed intake was the acid-base shift caused by the diets with AA substitution higher than 50 %. The dietary inclusion of Asn and Gln attenuated this acid-base shift. The knowledge gained from this dissertation can contribute to the targeted use of more free AA, representing a potential alternative to protein sources for the future and reducing the CP content of the feed. The free AA level used to date in practical feeding is below the limit identified in this dissertation. Nonetheless, such levels of free AA are often used in research to answer specific questions or to cover certain AA interactions for a more accurate estimation of the individual AA requirements for contemporary poultry breeds.
Modulation of hepatic insulin and glucagon signaling by nutritional factors in broiler chicken
(2022) Petrilla, Janka; Mátis, Gábor; Mackei, Máté; Kulcsár, Anna; Sebők, Csilla; Papp, Márton; Gálfi, Péter; Fébel, Hedvig; Huber, Korinna; Neogrády, Zsuzsanna
Influencing the endocrine metabolic regulation of chickens by nutritional factors might provide novel possibilities for improving animal health and productivity. This study was designed to evaluate the impact of dietary cereal type (wheat-based (WB) vs. maize-based (MB) diets), crude protein level (normal (NP) vs. lowered (LP)), and sodium (n-)butyrate (1.5 g/kg diet) supplementation (vs. no butyrate) on the responsiveness of hepatic glucagon receptor (GCGR), insulin receptor beta (IRβ) and mammalian target of rapamycin (mTOR) in the phase of intensive growth of chickens. Liver samples of Ross 308 broiler chickens (Gallus gallus domesticus) were collected on day 21 for quantitative real-time polymerase chain reaction and Western blot analyses. Hepatic GCGR and mTOR gene expressions were up-regulated by WB and LP diet. GCGR and IRβ protein level decreased in groups with butyrate supplementation; however, the quantity of IRβ and mTOR protein increased in WB groups. Based on these data, the applied dietary strategies may be useful tools to modulate hepatic insulin and glucagon signaling of chickens in the period of intensive growth. The obtained results might contribute to the better understanding of glycemic control of birds and increase the opportunity of ameliorating insulin sensitivity, hence, improving the production parameters and the welfare of broilers.
Dietary L-carnitine affects leukocyte count and function in dairy cows around parturition
(2022) Kononov, Susanne Ursula; Meyer, Jennifer; Frahm, Jana; Kersten, Susanne; Kluess, Jeannette; Bühler, Susanne; Wegerich, Anja; Rehage, Jürgen; Meyer, Ulrich; Huber, Korinna; Dänicke, Sven
In early lactation, an energy deficit leading to a negative energy balance (NEB) is associated with increased susceptibility to disease and has been shown to be an important factor during transition in dairy cows. L-carnitine as a key factor in the mitochondrial transport of fatty acids and subsequently for β-oxidation and energy release is known to modulate mitochondrial biogenesis and thus influence metabolism and immune system. In the current study, we characterized hematological changes around parturition and investigated the potential effects of dietary L-carnitine supplementation on immune cell functions. For this approach, dairy cows were assigned either to a control (CON, n = 30) or an L-carnitine group [CAR, n = 29, 25 g rumen-protected L-carnitine per cow and day (d)]. Blood samples were taken from d 42 ante partum (ap) until d 110 post-partum (pp), with special focus and frequent sampling from 0.5 to72 h post-calving to clarify the impact of L-carnitine supplementation on leukocyte count, formation of reactive oxygen species (ROS) in polymorphonuclear cells (PMN) and peripheral mononuclear cells (PBMC) and their phagocytosis activity. Blood cortisol concentration and the capacity of PBMC proliferation was also investigated. All populations of leukocytes were changed during the peripartal period, especially granulocytes showed a characteristic increase up to 4 h pp. L-carnitine supplementation resulted in increased levels of eosinophils which was particularly pronounced one day before to 4 h pp, indicating a possible enhanced support for tissue repair and recovery. Non-supplemented cows showed a higher phagocytic activity in PBMC as well as a higher phagocytic capacity of PMN during the most demanding period around parturition, which may relate to a decrease in plasma levels of non-esterified fatty acids reported previously. L-carnitine, on the other hand, led to an increased efficiency to form ROS in stimulated PMN. Finally, a short period around calving proved to be a sensitive period in which L-carnitine administration was effective.
Improving the accuracy of multi-breed prediction in admixed populations by accounting for the breed origin of haplotype segments
(2022) Schmid, Markus; Stock, Joana; Bennewitz, Jörn; Wellmann, Robin
Numerically small breeds have often been upgraded with mainstream breeds. This historic introgression predisposes the breeds for joint genomic evaluations with mainstream breeds. The linkage disequilibrium structure differs between breeds. The marker effects of a haplotype segment may, therefore, depend on the breed from which the haplotype segment originates. An appropriate method for genomic evaluation would account for this dependency. This study proposes a method for the computation of genomic breeding values for small admixed breeds that incorporate phenotypic and genomic information from large introgressed breeds by considering the breed origin of alleles (BOA) in the evaluation. The proposed BOA model classifies haplotype segments according to their origins and assumes different but correlated SNP effects for the different origins. The BOA model was compared in a simulation study to conventional within-breed genomic best linear unbiased prediction (GBLUP) and conventional multi-breed GBLUP models. The BOA model outperformed within-breed GBLUP as well as multi-breed GBLUP in most cases.
Multi-omics reveals different strategies in the immune and metabolic systems of high-yielding strains of laying hens
(2022) Iqbal, Muhammad Arsalan; Reyer, Henry; Oster, Michael; Hadlich, Frieder; Trakooljul, Nares; Perdomo-Sabogal, Alvaro; Schmucker, Sonja; Stefanski, Volker; Roth, Christoph; Camarinha Silva, Amélia; Huber, Korinna; Sommerfeld, Vera; Rodehutscord, Markus; Wimmers, Klaus; Ponsuksili, Siriluck
Lohmann Brown (LB) and Lohmann Selected Leghorn (LSL) are two commercially important laying hen strains due to their high egg production and excellent commercial suitability. The present study integrated multiple data sets along the genotype-phenotype map to better understand how the genetic background of the two strains influences their molecular pathways. In total, 71 individuals were analyzed (LB, n = 36; LSL, n = 35). Data sets include gut miRNA and mRNA transcriptome data, microbiota composition, immune cells, inositol phosphate metabolites, minerals, and hormones from different organs of the two hen strains. All complex data sets were pre-processed, normalized, and compatible with the mixOmics platform. The most discriminant features between two laying strains included 20 miRNAs, 20 mRNAs, 16 immune cells, 10 microbes, 11 phenotypic traits, and 16 metabolites. The expression of specific miRNAs and the abundance of immune cell types were related to the enrichment of immune pathways in the LSL strain. In contrast, more microbial taxa specific to the LB strain were identified, and the abundance of certain microbes strongly correlated with host gut transcripts enriched in immunological and metabolic pathways. Our findings indicate that both strains employ distinct inherent strategies to acquire and maintain their immune and metabolic systems under high-performance conditions. In addition, the study provides a new perspective on a view of the functional biodiversity that emerges during strain selection and contributes to the understanding of the role of host–gut interaction, including immune phenotype, microbiota, gut transcriptome, and metabolome.
Does carrying a rider change motor and sensory laterality in horses?
(2022) Schwarz, Sophie; Marr, Isabell; Farmer, Kate; Graf, Katja; Stefanski, Volker; Krueger, Konstanze
Laterality in horses has been studied in recent decades. Although most horses are kept for riding purposes, there has been almost no research on how laterality may be affected by carrying a rider. In this study, 23 horses were tested for lateral preferences, both with and without a rider, in three different experiments. The rider gave minimal aids and rode on a long rein to allow the horse free choice. Firstly, motor laterality was assessed by observing forelimb preference when stepping over a pole. Secondly, sensory laterality was assessed by observing perceptual side preferences when the horse was confronted with (a) an unfamiliar person or (b) a novel object. After applying a generalised linear model, this preliminary study found that a rider increased the strength of motor laterality (p = 0.01) but did not affect sensory laterality (p = 0.8). This suggests that carrying a rider who is as passive as possible does not have an adverse effect on a horse’s stress levels and mental state.
Central carbon metabolism, sodium-motive electron ransfer, and ammonium formation by the vaginal pathogen Prevotella bivia
(2021) Schleicher, Lena; Herdan, Sebastian; Fritz, Günter; Trautmann, Andrej; Seifert, Jana; Steuber, Julia
Replacement of the Lactobacillus dominated vaginal microbiome by a mixed bacterial population including Prevotella bivia is associated with bacterial vaginosis (BV). To understand the impact of P. bivia on this microbiome, its growth requirements and mode of energy production were studied. Anoxic growth with glucose depended on CO2 and resulted in succinate formation, indicating phosphoenolpyruvate carboxylation and fumarate reduction as critical steps. The reductive branch of fermentation relied on two highly active, membrane-bound enzymes, namely the quinol:fumarate reductase (QFR) and Na+-translocating NADH:quinone oxidoreductase (NQR). Both enzymes were characterized by activity measurements, in-gel fluorography, and VIS difference spectroscopy, and the Na+-dependent build-up of a transmembrane voltage was demonstrated. NQR is a potential drug target for BV treatment since it is neither found in humans nor in Lactobacillus. In P. bivia, the highly active enzymes L-asparaginase and aspartate ammonia lyase catalyze the conversion of asparagine to the electron acceptor fumarate. However, the by-product ammonium is highly toxic. It has been proposed that P. bivia depends on ammonium-utilizing Gardnerella vaginalis, another typical pathogen associated with BV, and provides key nutrients to it. The product pattern of P. bivia growing on glucose in the presence of mixed amino acids substantiates this notion.
Na+-coupled respiration and reshaping of extracellular polysaccharide layer counteract monensin-induced cation permeability in Prevotella bryantii B14
(2021) Trautmann, Andrej; Schleicher, Lena; Pfirrmann, Jana; Boldt, Christin; Steuber, Julia; Seifert, Jana
Monensin is an ionophore for monovalent cations, which is frequently used to prevent ketosis and to enhance performance in dairy cows. Studies have shown the rumen bacteria Prevotella bryantii B14 being less affected by monensin. The present study aimed to reveal more information about the respective molecular mechanisms in P. bryantii, as there is still a lack of knowledge about defense mechanisms against monensin. Cell growth experiments applying increasing concentrations of monensin and incubations up to 72 h were done. Harvested cells were used for label-free quantitative proteomics, enzyme activity measurements, quantification of intracellular sodium and extracellular glucose concentrations and fluorescence microscopy. Our findings confirmed an active cell growth and fermentation activity of P. bryantii B14 despite monensin concentrations up to 60 µM. An elevated abundance and activity of the Na+-translocating NADH:quinone oxidoreductase counteracted sodium influx caused by monensin. Cell membranes and extracellular polysaccharides were highly influenced by monensin indicated by a reduced number of outer membrane proteins, an increased number of certain glucoside hydrolases and an elevated concentration of extracellular glucose. Thus, a reconstruction of extracellular polysaccharides in P. bryantii in response to monensin is proposed, which is expected to have a negative impact on the substrate binding capacities of this rumen bacterium.
Electronic nose for the rapid detection of deoxynivalenol in wheat using classification and regression trees
(2022) Camardo Leggieri, Marco; Mazzoni, Marco; Bertuzzi, Terenzio; Moschini, Maurizio; Prandini, Aldo; Battilani, Paola
Mycotoxin represents a significant concern for the safety of food and feed products, and wheat represents one of the most susceptible crops. To manage this issue, fast, reliable, and low-cost test methods are needed for regulated mycotoxins. This study aimed to assess the potential use of the electronic nose for the early identification of wheat samples contaminated with deoxynivalenol (DON) above a fixed threshold. A total of 214 wheat samples were collected from commercial fields in northern Italy during the periods 2014–2015 and 2017–2018 and analyzed for DON contamination with a conventional method (GC-MS) and using a portable e-nose “AIR PEN 3” (Airsense Analytics GmbH, Schwerin, Germany), equipped with 10 metal oxide sensors for different categories of volatile substances. The Machine Learning approach “Classification and regression trees” (CART) was used to categorize samples according to four DON contamination thresholds (1750, 1250, 750, and 500 μg/kg). Overall, this process yielded an accuracy of >83% (correct prediction of DON levels in wheat samples). These findings suggest that the e-nose combined with CART can be an effective quick method to distinguish between compliant and DON-contaminated wheat lots. Further validation including more samples above the legal limits is desirable before concluding the validity of the method.
The active core microbiota of two high-yielding laying hen breeds fed with different levels of calcium and phosphorus
(2022) Roth, Christoph; Sims, Tanja; Rodehutscord, Markus; Seifert, Jana; Camarinha-Silva, Amélia
The nutrient availability and supplementation of dietary phosphorus (P) and calcium (Ca) in avian feed, especially in laying hens, plays a vital role in phytase degradation and mineral utilization during the laying phase. The required concentration of P and Ca peaks during the laying phase, and the direct interaction between Ca and P concentration shrinks the availability of both supplements in the feed. Our goal was to characterize the active microbiota of the entire gastrointestinal tract (GIT) (crop, gizzard, duodenum, ileum, caeca), including digesta- and mucosa-associated communities of two contrasting high-yielding breeds of laying hens (Lohmann Brown Classic, LB; Lohmann LSL-Classic, LSL) under different P and Ca supplementation levels. Statistical significances were observed for breed, GIT section, Ca, and the interaction of GIT section x breed, P x Ca, Ca x breed and P x Ca x breed (p < 0.05). A core microbiota of five species was detected in more than 97% of all samples. They were represented by an uncl. Lactobacillus (average relative abundance (av. abu.) 12.1%), Lactobacillus helveticus (av. abu. 10.8%), Megamonas funiformis (av. abu. 6.8%), Ligilactobacillus salivarius (av. abu. 4.5%), and an uncl. Fusicatenibacter (av. abu. 1.1%). Our findings indicated that Ca and P supplementation levels 20% below the recommendation have a minor effect on the microbiota compared to the strong impact of the bird’s genetic background. Moreover, a core active microbiota across the GIT of two high-yielding laying hen breeds was revealed for the first time.
Feed clusters according to In Situ and In Vitro ruminal crude protein degradation
(2023) Okon, Paul; Bachmann, Martin; Wensch-Dorendorf, Monika; Titze, Natascha; Rodehutscord, Markus; Rupp, Christiane; Susenbeth, Andreas; Greef, Jörg Michael; Zeyner, Annette
Effective degradation (ED) of crude protein (CP) was estimated in vitro at 0.02, 0.05 and 0.08 h−1 assumed ruminal passage rates for a total of 40 feedstuffs, for which in situ ED was available and used as reference degradation values. For this, the Streptomyces griseus protease test was used. The differences between in vitro CP degradation and the in situ CP degradation values were lowest in legume grains and highest in cereal by-products and barley. The differences between in situ and in vitro ED were expressed using a degradation quotient (degQ), where degQ = (EDin vitro − EDin situ)/EDin situ. Among the tested feedstuffs, eight specific clusters were identified according to degQ for the assumed passage rates. The feedstuffs clustered in an unspecific way, i.e., feedstuffs of different nutrient composition, origin or treatment did not necessarily group together. Formaldehyde–treated rapeseed meal, soybean meal, wheat, a treated lupin, sunflower meal and barley could not be assigned to any of the clusters. Groupwise degradation (range of degQ for assumed passage rates are given in brackets) was detected in grass silages (−0.17, −0.11), cereal by-products together with sugar beet pulp (−0.47, −0.35) and partly in legume grains (−0.14, 0.14). The clustering probably based on different specific nutrient composition and matrix effects that influence the solubility of feed protein and limit the performance of the protease. The matrix can be affected by treatment (chemically, thermally or mechanically), changing the chemical and physical structure of the protein within the plant. The S. griseus protease test had reliable sensitivity to reflect differences between native feedstuffs and treatments (thermally or chemically) that were found in situ. The in situ results, however, are mostly underestimated. The clustering results do not allow a clear conclusion on the groupwise or feed-specific use of carbohydrate-degrading enzymes as pre- or co-inoculants as part of the S. griseus protease test and need to be tested for its potential to make this test more conform with in situ data.
Bi-objective optimization of nutrient intake and performance of broiler chickens using Gaussian process regression and genetic algorithm
(2023) Ahmadi, Hamed; Rodehutscord, Markus; Siegert, Wolfgang
This study investigated whether quantifying the trade-off between the maxima of two response traits increases the accuracy of diet formulation. To achieve this, average daily weight gain (ADG) and gain:feed ratio (G:F) responses of 7–21-day-old broiler chickens to the dietary supply of three nutrients (intake of digestible glycine equivalents, digestible threonine, and total choline) were modeled using a newly developed hybrid machine learning-based method of Gaussian process regression and genetic algorithm. The dataset comprised 90 data lines. Model-fit-criteria indicated a high model adjustment and no prediction bias of the models. The bi-objective optimization scenarios through Pareto front revealed the trade-off between maximized ADG and maximized G:F and provided information on the needed input of the three nutrients that interact with each other to achieve the trade-off scenarios. The trade-off scenarios followed a nonlinear pattern. This indicated that choosing target values intermediate to maximized ADG and G:F after single-objective optimization is less accurate than feed formulation after quantifying the trade-off. In conclusion, knowledge of the trade-off between maximized ADG and maximized G:F and the needed nutrient inputs will help feed formulators to optimize their feed with a more holistic approach.
Update on shedding and transmission routes of porcine haemotrophic mycoplasmas in naturally and experimentally infected pigs
(2021) Ade, Julia; Ritzmann, Mathias; Wöstmann, Christopher; Eddicks, Matthias; Reese, Sven; Hoelzle, Katharina; Hoelzle, Ludwig E.; Stadler, Julia
Horizontal transmission of Mycoplasma suis via parenteral exposure during standard practices or through bites during fightings have been identified as key epidemiological routes. However, as knowledge gaps on other potential shedding and transmission routes exist, the present study combines both laboratory experiments and field surveys to gain new insights into the epidemiology of porcine haemotrophic mycoplasmas. Splenectomised pigs were orally inoculated with a M. suis field strain and investigated for clinical signs related to infectious anaemia of pigs (IAP) and the presence of M. suis in blood, urine and saliva samples by qPCR. All blood samples were negative for M. suis and animals did not show obvious clinical signs of IAP throughout the entire study period. Additionally, urine, nasal and saliva samples from sows of conventional piglet producing farms and semen samples from a boar stud revealed no detection of M. suis and ‘Candidatus Mycoplasma haemosuis’ by qPCR. Thus, the results indicate that blood-independent transmission routes might be of minor relevance under field conditions.
Clinical, haematological and pathomorphological findings in Mycoplasma suis infected pigs
(2021) Stadler, Julia; Ade, Julia; Hermanns, Walter; Ritzmann, Mathias; Wentzel, Sarah; Hoelzle, Katharina; Hoelzle, Ludwig E.
Background: Mycoplasma suis (M. suis) belongs to the group of haemotrophic mycoplasmas and is known as the causative agent of infectious anaemia in pigs. In the last few years valuable insights into the mechanism of adhesion and invasion, shedding patterns and cell tropism of M. suis were gained by the use of new molecular techniques. However, details on M. suis induced lesions as well as the distribution of M. suis in different organs are still lacking. Therefore, seven splenectomised pigs were experimentally infected and clinical and laboratory investigations as well as a detailed histopathological examination were performed. Detection and quantification of M. suis DNA in blood and various tissue samples was done using a quantitative real-time PCR. Results: During the course of experimental infection, periodically occurring signs of infectious anaemia of pigs including severe icteroanaemia, fever, apathy and anorexia were observed. In addition, dermatological manifestations such as haemorrhagic diathesis presenting as petechiae occurred. The most important haematological alterations were normochromic, normocytic anaemia, hypoglycaemia as well as increased bilirubin and urea concentrations. Necropsy revealed predominant evidence of haemolysis with consecutive anaemia, as well as disseminated intravascular coagulation. M. suis was found in all investigated tissues with the highest copy numbers found in the kidneys. In Giemsa stained sections M. suis was only detected red blood cell (RBC)-associated. Conclusion: In the present study, no RBC independent sequestration of M. suis was detected in organs of experimentally infected pigs. Pathological findings are most likely resulting from haemolysis, consecutive anaemia as well as from disseminated intravascular coagulation and subsequent organ impairments.

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