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Publication The adoption potential of extended lactation as a strategy to reduce excess calf numbers in dairy farming(2024) Gresham, Josephine; Reiber, Christoph; Chagunda, Mizeck G. G.Dairy production relies predominantly on a few, selectively bred, high-yielding dairy breeds, resulting in a surplus of low-value male offspring. This situation leads to a conflict between a growing demand for milk and low appreciation for dairy calves. Extending lactation could reduce the number of calves born. This study aimed to assess the current use of extended lactation in dairy production in Southern Germany, identify the perceived potential, and determine the biological potential extended lactation has to reduce calf numbers. A survey from 2020 with 310 farmers was analysed. A total of 145 (46.8%) farmers used extended lactation. The use of extended lactation on farms correlated significantly with breed, milk yield, interest in continuing or implementing the use of extended lactation, and the indicated feasibility of extended lactation. The perceived potential was assessed by 39.8% of farmers as “high” to “very high”. A total of 17 (12.8%) farmers currently not using extended lactation were identified as potential adopters. Together with existing adopters, this could result in 52.3% of farms using extended lactation, which could biologically reduce calf numbers by 7.3% or 14.1% p.a., when extended by three or six months, respectively. This adoption potential shows that extended lactation can reduce calf numbers considerably, addressing both ethical and economic concerns in dairy farming and benefiting farmers as well as society.Publication Degradation of crude protein and starch of corn and wheat grains in the rumen(2016) Seifried, Natascha; Rodehutscord, MarkusThe major objectives of the present thesis were to characterize the ruminal crude protein (CP) and starch degradation of different genotypes of corn and wheat grains and to predict the effective degradation (ED) of CP and starch with easily measurable characteristics. The in situ method is the standard technique to study the ruminal degradation of feeds in many feed evaluation systems. This technique was originally applied to study forages and it was therefore necessary to clarify methodical details related to the measurements of in situ starch degradation from cereal grains. Two in vitro and one in situ approach were conducted to study the loss of secondary starch particles from bags with different pore sizes used for the in situ incubation of different cereal grains. In the first in vitro study ground wheat was incubated in bags (pore size: 50 µm) over different time spans in a modified rumen simulation technique. Bag residues and fermenter fluids were analyzed for their starch content. In the second in vitro study ground wheat, barley, and corn were incubated with bags of pore sizes of 50, 30 (except corn), 20, and 6 µm. In the in situ study ground wheat, barley, corn, and oats were rumen incubated over different time spans using bags with pore sizes of 50, 20, and 6 µm. The starch content of the grains and bag residues was analyzed enzymatically and the degradation characteristics of starch were calculated for each grain type and pore size. It was shown for the first time that incubating wheat and barley in bags with 50 and 30 µm pore size lead to a substantial amount of secondary starch particle losses during incubation process in vitro. These losses were not detectable when the grains were incubated with bags having pore sizes of 20 and 6 µm. Independent of the bags’ pore size no secondary starch particle losses were found by the incubation of corn. Thus corn can be studied in situ even with bags with 50 µm pore size. Oats showed very high washout losses with all pore sizes tested in the present thesis and therefore none of them is suitable to study the starch degradation measurements of oats. Because of methodical problems of gas accumulation in bags having pore sizes < 50 µm, no recommendations can be provided for the in situ evaluation of wheat and barley. Further research is necessary to solve these problems. In the second and third study of the present thesis ruminal in situ degradation of 20 corn grain genotypes and 20 wheat grain genotypes was measured in three lactating Jersey cows. In both experiments the same techniques were used to characterize the ruminal degradation of CP and starch. Ground grains (2 mm) were rumen incubated in bags (50 µm pore size) over different time spans. Grains and bag residues were analyzed for their CP and starch content. The degradation parameters and the ED were calculated for dry matter (DM), CP, and starch. Gas production (GP) of ground grains (1 mm) was recorded after incubation over different time spans in buffered ruminal fluid and fitted to an exponential equation to determine GP parameters. To predict ED of CP and starch correlations with physical and chemical characteristics and in vitro measurements were evaluated and stepwise multiple linear regression analyses were applied. The in situ parameters (soluble fraction, potential degradable fraction, and degradation rate) varied widely between genotypes of corn and wheat grains. The ED of DM, CP, and starch showed a high variation for corn grain genotypes. Due to the high degradation rates, the ED of wheat grains were similar between genotypes. The GP rate was in good agreement with the in situ values for corn grains, whereas no systematic relationship between both methods was observed for wheat grains. Evaluation of correlation analysis showed significant relationships between calculated ED of CP and several amino acids (AA) for both grain types. This indicates that the protein composition of the grains influences CP degradation in the rumen. Similar relationships were found between the same AA and ED of starch of corn grains which highlights the impact of the protein composition on ruminal starch degradation for this grain type. For both grain types, the ED of starch and CP could be predicted accurately from physical and chemical characteristics alone or in combination with GP measurements. Thus, the equations presented in the present thesis can be used to obtain rapid and cost effective information on ruminal degradation of CP and starch for corn and wheat grains. The results of the present thesis show that there is considerable variation of ruminal CP and starch degradation from different genotypes of corn and also – albeit to a lesser extent – for wheat grains. Differences in ED of starch should be taken into account when formulating rations containing significant amounts of corn and wheat grains. In the case of corn grains differences in ED of CP should also be accounted for.Publication 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, SvenIn 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.Publication Dietary L-carnitine supplementation modifies blood parameters of mid-lactating dairy cows during standardized lipopolysaccharide-induced inflammation(2024) Seemann, Leonie; Frahm, Jana; Kersten, Susanne; Bühler, Susanne; Meyer, Ulrich; Visscher, Christian; Huber, Korinna; Dänicke, SvenL-carnitine, available as feed additive, is essential for the beta-oxidation of free fatty acids in the mitochondrial matrix. It provides energy to immune cells and may positively impact the functionality of leukocytes during the acute phase response, a situation of high energy demand. To test this hypothesis, German Holstein cows were assigned to a control group (CON, n = 26) and an L-carnitine supplemented group (CAR, n = 27, rumen-protected L-carnitine product: 125 g/cow/d, corresponded to total L-carnitine intake: 25 g/cow/d, supplied with concentrate) and received an intravenous bolus injection of lipopolysaccharides (LPS, 0.5 µg/kg body weight, E. coli) on day 111 postpartum as a model of standardized systemic inflammation. Blood samples were collected from day 1 ante injectionem until day 14 post injectionem (pi), with frequent sampling through an indwelling venous catheter from 0.5 h pi to 12 h pi. All parameters of the white blood cell count responded significantly to LPS, while only a few parameters were affected by L-carnitine supplementation. The mean eosinophil count, as well as the percentage of basophils were significantly higher in CAR than in CON over time, which may be due to an increased membrane stability. However, phagocytosis and production of reactive oxygen species by leukocytes remained unchanged following L-carnitine supplementation. In conclusion, although supplementation with 25 g L-carnitine per cow and day resulted in increased proportions of specific leukocyte populations, it had only minor effects on the functional parameters studied in mid-lactating dairy cows during LPS-induced inflammation, and there was no evidence of direct improvement of immune functionality.Publication Evaluation of a UHF RFID system for livestock(2017) Hammer, Nora; Jungbluth, ThomasA structural change could be observed within German animal production in recent years. Whereas the number of livestock holdings decreased, the number of animals per livestock holding increased. Because bigger livestock holdings are also often in a conflict of aims between sustainability, animal welfare and economy, a well-functioning and cost-effective management assistance is even more important. The collection of animal-related data and data from their environment with simple, innovative and low-cost techniques to improve animal welfare, animal health and animal performance, as well as the housing conditions, is a main part of so-called precision livestock farming (PLF). A possible solution for implementation of these thoughts is a technology called radio-frequency identification (RFID). The suitability of an UHF RFID system for simultaneous pig and cattle detection could be evaluated during a three year project, which was funded by the Federal Office of Agriculture and Food. Therefore, several UHF transponder ear tags had to be developed and tested in laboratory and practical experiments. Additionally, a cost-benefit analysis of the UHF system based on four example barns had to be carried out to estimate the potential of the system for use in practice. Thereby, not only the costs and benefits of simultaneous animal detection were calculated, but also the costs and benefits of hotspot monitoring of the animals in their husbandry environment were estimated. Nine different transponder types for each animal species were developed within the duration of the project. During the development process, the antenna structure, antenna length and label material had to be varied to adjust the transponder to its immediate surroundings as optimally as possible. The grouting process of the transponder into the ear tag was also continuously improved. Before testing the UHF transponder ear tag types in practice, they were all tested on a dynamic test bench. Using this test bench, a preliminarily assessment of the in-house developed transponder types by themselves, with foreign and commercially available UHF transponder types under various conditions was possible. The number of readings per round was recorded and used to identify differences between the transponder types. The UHF transponder ear tag types were tested with the aid of driving experiments using pigs and cattle with a focus on their suitability and durability under practical conditions. While one gate in a stall environment was built in the driving experiments for the fattening pigs, with cattle, reader output power, reader orientation and the test environment were varied. In these experiments, the number of readings per round and the reading rates, which were the more decisive value in practice, were calculated. In the last stage of development, a suitable, well-functioning UHF transponder ear tag type and good average reading rates could be achieved for both animal species (pigs: ø 98 %; cattle: ø 99 %) While performing the cost-benefit analysis, it could be calculated that, at the present state of development of the UHF system, the benefits do not exceed the costs of the system in the fattening pig husbandry. In dairy cattle husbandry, a positive result could be reached only under the best estimations and the larger farm. However, the costs arising per animal are still too high to implement the systems on the market. Because of the early stage of development, the calculation of the costs and benefits was difficult and still holds uncertainties. Following the assumption that the UHF system will be developed to practical maturity, the costs calculated would be lower and an advantageousness of the system would be also expected for other farms. This work was sensible and necessary to get a first assessment of the costs and benefits. Great development progress could be achieved for the UHF RFID system and a large potential for PLF could be shown within this project, even if the system is not yet ready for market.Publication Impact of physically effective fiber on protein metabolism in dairy cows(2022) Heering, Ruth; Dickhöfer, UtaFiber is a prerequisite in the nutrition of dairy cows, because it promotes rumen motility and mat formation, while stimulating chewing activity (i.e. eating and ruminating). Chewing and salivation ensure a stable rumen environment for the microbial symbionts by providing an even release and recycling of nutrients as well as by buffering the end products of rumen fermentation, and thus reducing the risk of acidosis. The concept of physically effective neutral detergent fiber (peNDF) has been introduced to evaluate the adequacy of fiber supply to dairy cows. The peNDF combines both, the physical and the chemical characteristics of fiber, namely the dietary particle size and the neutral detergent fiber concentration. Various studies have been conducted to understand the effects of peNDF, particularly those concerning rumen health issues and the prevention of acidosis. Although some studies have looked at the effect of peNDF on nitrogen (N) metabolism, no studies are known that have evaluated the impact of peNDF on the partitioning of N excretion in dairy cows. Given that the peNDF can be considered as that fraction of the feed that stimulates chewing and salivation in ruminants, it may enhance recycling of circulating N and therefore increase the efficiency of N use by the animal. In this doctoral project, it was hypothesized that increasing the peNDF concentration of a diet stimulates chewing activity in dairy cows, thereby promoting salivation up to a certain peNDF concentration, after which chewing activity declines as a result of lower dry matter (DM) intake. It was further hypothesized that greater chewing activity and salivation may stimulate rumen N recycling and microbial protein synthesis (MPS), compensating for potential negative effects of reduced rumen-degradable crude protein supply on feed intake, nutrient digestibility, and performance of dairy cows. The present dissertation uses the rumen N balance (RNB) to evaluate the availability of degradable crude protein in the rumen. Overall, three in vivo studies were conducted to investigate the proposed hypotheses. The first study tested the effects of four graded peNDF concentrations of a total mixed ration (TMR) on the chewing activity of eight lactating dairy cows to evaluate whether the selected peNDF concentrations have an effect on chewing activity. The second study evaluated the effects of different combinations of two dietary peNDF concentrations and two RNB (RNB0: 0 g/kg DM and RNB–: – 1.5 g/kg DM) in a TMR fed to twenty lactating dairy cows. The third study tested the effects of four different peNDF concentrations in a TMR fed to four rumen-fistulated lactating dairy cows. Overall, increasing the peNDF concentration did not always result in a reduction of DM intake by cows, which could be related to their stage of lactation and energy requirement. Total chewing time and total chews per kilogram of DM intake increased linearly with increasing peNDF concentration; however, the increment in chewing activity may have resulted only in a small increase in salivation and hence, in N recycling in the animal. Although, high peNDF concentration negatively affected the yield and efficiency of MPS and apparent total tract digestibility (ATTD) of nutrients at low RNB, there were no indications that these parameters had any effect on the yield or composition of milk produced by the cows. There was also an interaction between the peNDF concentration and the RNB in the diets of dairy cows, with the effects being more pronounced at low RNB. Therefore, recommended peNDF concentration for dairy cows will vary with RNB and vice versa. The minimum RNB needed to sustain rumen functioning may differ depending on the dietary peNDF concentration. In this regard, more studies are needed for a better understanding of the interaction between RNB and peNDF concentration. Finally, offering dairy cows a slightly negative RNB diet at an optimum peNDF concentration may be a means of reducing the N emission from dairy milk production and hence, may provide economical and environmental benefits.Publication Langfristige Beurteilung der Energieversorgung von Milchkühen bei unterschiedlichem Kraftfuttereinsatz(2020) Gerster, Elisabeth Katharina; Rodehutscord, MarkusThe first objective of the present study was to compile the current knowledge concerning the effect of the amount of CON on dry matter (DM) intake and milk yield of dairy cows, by regression analysis. For this purpose, a dataset was compiled containing 46 studies with feeding trials and graded inclusion levels of CON. When offering the roughage ad libitum, a quadratic relation was found between CON intake and roughage intake. Taking into account the substitution of roughage, the observed increase of milk yield was less than expected. The second objective was to verify the response of Simmental dairy cows to a long-term reduction of the CON input, in terms of dry matter intake, milk yield, and energy supply. During a feeding trial conducted over a two-year period at the experimental station LAZBW in Aulendorf (cooperative research project „optiKuh“) each 24 Simmental dairy cow was fed either 250 (group 250) or 150 (group 150) g CON per kg energy-corrected milk yield (ECM) throughout lactation. Grass and maize silage as well as straw and hay were combined in the total mixed rations to ensure an energy concentration of 6.6 MJ NEL per kg DM of the roughage in both groups throughout lactation. The CON contained a mixture produced on-site (winter wheat, winter barley, faba bean) and rapeseed meal. At 165 days in milk, each cow in both groups was switched individually to a ration containing less CON. Data were gathered also during the dry period. Feed was offered for ad libitum consumption. Statistical analyses were done separately for the lactation and dry period. As fixed effects the calendar week, the group, the parity, and the interaction group*parity entered the model, respectively. Additionally, the days in milk were included as a covariate during lactation. During lactation the mean daily DM intake did not differ significantly between group 250 (21.6 kg DM) and group 150 (21.0 kg DM). The calculated substitution of roughage amounted to 0.75 kg DM per kg CON DM during the mentioned trial period. As lactation progressed, more roughage was substituted. The group 250 had a higher milk yield during the first half of lactation, but a lower milk yield during the second half. The mean milk yield was equal in both groups (28.7 kg ECM per day). The examined blood characteristics (8, 28, 100 days p. p., at drying off, 14 days a. p.) and the milk fatty acids analysis by gas chromatography (5, 28, 100, 200 days p. p.) reflected the different progressions of the energy balance of the two groups. On average the group 250 reached a positive energy balance already at 37 days in milk. Probably owing to a better glucose supply after calving, the serum insulin concentration increased faster and supported the replenishing of body reserves at the expense of milk yield. On the contrary, the group 150 achieved a positive energy balance only after 72 days in milk on average. Indicating the mobilization of body reserves, the blood of group 150 showed a significantly higher D-ß-hydroxybutyrate concentration, and the milk fat a significantly higher concentration of long-chain fatty acids 28 days p.p.. However, the values were within the physiological range. The reduction of CON did not increase the likelihood of illness. During the dry period, despite equal feeding of both groups in this period, the group 150 had an increased DM intake of 0.9 kg DM per day (p=0,026). Thus, they restored body energy reserves. Over time both groups succeeded in balancing the energy deficit of the beginning of lactation. The proposal of SUSENBETH (2018) for the calculation of the energy requirement of dairy cows improved the plausibility of the calculated energy balances compared with the factors according to GfE (2001). The third objective was to develop a model for the estimation of energy balance in the first third of lactation based on characteristics of production with focus on concentration of milk fatty acids. For this purpose, the milk fatty acids analyses 5, 28 and 100 days p. p. of the feeding trial were used (n=200). The model was derived from the regularized linear regression method „elastic net“. The variables milk yield on sampling day, milk fat content, concentrations of the milk fatty acid C12:0 and the milk fatty acid C18:0, as well as the relations n-6/n-3, C15/C17 and oleic acid/C15 were selected. An estimation error of 13.1 MJ NEL per day was determined by leave-one-out cross-validation. Potentially, the model seems to be applicable for the detection of a severe negative energy balance at the beginning of lactation. But first a validation with an independent dataset is required.Publication Prediction of ruminal acidosis in dairy cows from milk constituents(2022) Seyfang, Gero Marc; Rodehutscord, MarkusSubacute rumen acidosis (SARA) is a common, but hardly assumable disease in modern dairy cows’ herds. SARA incidences are prevalent in two circumstances. The first, when the cows have to adapt fast to a ration high in carbohydrates after parturition. Since the feed composition has to be changed fast, to meet the cows’ requirements energy- and nutrients wise, the rumen microbiota climate has to adapt fast, which can cause unbeneficial rumen circumstances. The second, when the lactating cows have, beside high milk yield also a high feed intake in mid-lactation, when feed high in energy but low in structural carbohydrates is fed. This can lead to high density of VFAs in the rumen, if the outflow and absorption through the ruminal wall, as well as the buffer capacity in the rumen is not sufficient for the high production of those acids. Then the ruminal milieu becomes more acid, which can negatively affect the cow’s health. The cows suffering SARA, if at all, show mild symptoms like reduced water and feed intake, depression, diarrhea, reduced rumen motility, laminitis or reduced milk yield and milk fat depression. Since those symptoms can also show up with a delay in time and can be caused by several other factors, monitoring SARA in herds can be difficult. An unambiguous definition of SARA circumstances in the rumen cannot be found in literature, although it is under research for decades. Since SARA can influence the milk yield and can lead to a milkfat depression and a change in composition of milkfat, we focused on milk parameters and milkfat composition in particular with the aim of correlating those with pH conditions in the rumen. Three trials were made with feeding rations that were predictably capable of inducing SARA conditions in mid-lactation. During the trials, besides performance and ruminal parameters, as well as continuous pH measurement, milk samples were taken. The cows used were all rumen cannulated. Therefore, datalogger with integrated pH meter (Large Ruminant Logger M5-T7, Dascor Inc., Oceanside, USA) were placed in the ventral sac of the rumen to measure reliably and continuously. In Trial 1, three feeding rations with constant 20% grass silage were used. One ration consisted of additionally 20% corn silage and 60% concentrate (treatment CS60), the other two rations included 20 respectively 60% pressed sugar beet pulp silage and 60 respectively 20% concentrate (treatments SBPS60 respectively SBPS20). With those rations, low pH values were induced in the rumen, leading to SARA incidences of 89% in the measured days in the CS60, 100% in the SBPS60, and 61% in the SBPS20 treatment. In Trial 2, for all three rations a fix concentration of 52% concentrate was used. The remaining 48% consisted of corn silage (treatment CS), grass silage (treatment GS) or hay (treatment Hay). In the CS treatment, SARA incidence was 23%, while the GS and Hay treatments did not show SARA incidence. While the first two trials were designed as a 3x3 Latin square, in Trial 3 the cows remained in their respective treatment. One group stayed in the barn with a TMR, including 30% concentrate (treatment CG), while the other group was full time grazing and got additional 1.75 kg concentrate per day (treatment PG). SARA incidences were 7% in the CG and 8% in the PG. Additionally, in an intertrial approach, regression models for SARA detection were developed. Therefore, easily accessible performance data from the barn and milk parameters from the official milk control and milk fatty acids were used to estimate the rumen parameters pH mean and the time spent below pH 5.8. One first model was designed to include 63 variables. Besides 11 parameters gained in the barn or from the official milk control, also 52 parameters that were gaschromatographically detected fatty acids and sums of these fatty acids. A second model was designed to be useable if no gaschromatographical milkfat analysis was available. Therefore, only those FAs were included that can be estimated in a good quality with MIR spectroscopy. With those regression models the SARA days from the 185 measurement days were calculated to test the accuracy of the models. From the original 47 SARA days the first model was able to detect 43 days and the second model detected 39 SARA days. Although the accuracy of SARA prediction based on these models might be too inaccurate for a decision if a single day was SARA prevalent or not, an information on herd basis seems assessable. Still the small number of cows and measured days, as well as the fact that two breeds of cows and only cows in the later lactation phase were integrated in the model establishment has to be considered and further developed before it becomes a useful tool in field use for SARA detection.Publication The influence of L-carnitine on hematology and functional blood parameters of dairy cows with special focus on high resolution data around parturition(2023) Kononov, Susanne Ursula; Huber, KorinnaThe transition period, defined as three weeks before to three weeks after parturition, is one of the most critical times in the production cycle of dairy cows. On the one hand, cows have to cope with increased energy demand, while on the other hand, feed intake decreases due to stress and pain during parturition. This results in an negative energy balance and, consequently, at the beginning of body fat tissue mobilization. Lipomobilization increases the blood concentration of NEFA. This is accompanied by an increase in the blood concentration of ketone bodies, such as BHB. In addition to changes in energy metabolism, alterations in the immune function of dairy cows occur during the transition period. Stress and pain during calving lead to elevated blood levels of glucocorticoids, such as cortisol, which affect the immune system. Furthermore, the immune system is affected by increased concentrations of NEFA and BHB. At the same time, oxidative stress occurs due to an imbalance between the production of reactive oxygen species (ROS) and the activity of the antioxidative system. In general, the period around calving and its consequences constitute a very complex process influenced by many interdependent factors. One key factor in energy production is the quaternary amine L-carnitine (LC), which is necessary for the transport of short-chain fatty acids from the cytosol to the mitochondrial matrix. Furthermore, several studies have demonstrated the antioxidant and membrane-stabilizing effects of LC. This study aimed to investigate the effects of dietary LC supplementation on energy metabolism, hematology, and immune functions of dairy cows during the transition period. In addition, the first 72 h after calving were observed at high resolution to show the characteristic courses of the examined parameters, which, to the best of our knowledge, have not yet been analyzed. To attain this aim, 60 pluriparous Holstein Friesian cows were assigned to two groups based on their lactation number, body weight, body condition score, and fat-corrected milk yield from previous lactation. The LC group (CAR) received 25 g of rumen-protected LC. The study started 42 days before excepted calving and ended 110 days after parturition. To evaluate the performance and health of the animals, feed and milk samples were collected regularly, and feed intake, milk yield, body weight, and BCS were documented (Manuscript I). Additionally, NEB was calculated, and NEFA, BHB, and triglyceride concentrations in the blood were determined (Manuscript I). Also, the concentration of LC in the blood as well as that of the precursors γ-butyrobetaine (γBB), Nε-trimethyllysine (TML), and acetylcarnitine (ACA) was examined (Manuscript I). Red blood cell counts and antioxidant enzyme activity were measured to obtain more information on the oxygen supply and antioxidant status of the animals (Manuscript II). To evaluate the immunological status and inflammatory response, white blood cell count, phagocytic activity, ROS production, and lymphocyte populations were analyzed (Manuscript III). Dietary supplementation with LC increases blood LC, γBB, and ACA concentrations. Furthermore, LC supplementation resulted in better utilization of NEFA and TG. This was manifested by an increased blood concentration of triglycerides and a lower concentration of NEFA. Moreover, increased levels of platelets and eosinophils were detected in the CAR group, confirming the membrane-stabilizing effect of LC and the associated longer cell lifespan. Additionally, immunological functions were affected by LC supplementation. The ability of polymorphonuclear cells (PMN) to phagocytose bacteria was analyzed by the mean fluorescence intensity (MFI) of ROS-producing PMN, and the phagocytic capacity decreased compared to the CON group. Simultaneously, the efficiency of ROS production by PMN increased in CAR cows. These results suggest an altered immune function around calving, but not suppression, as is often described in the literature. In addition, this study showed that calving affected almost all analyzed data. The strongest changes in hematology and cell function were found four hours after calving. Furthermore, the influence of LC supplementation on immunological parameters was observed in the first few hours after parturition, indicating that LC supplementation may have an effect at energetically critical times. In conclusion, the present study showed that dietary LC supplementation affected energy metabolism, cell vitality, and cell function during the critical period around calving. However, this study also showed the clear influences of calving, which may be even more pronounced than animal-specific differences. Future studies should record the LC supply of cells to enable a more detailed description of the energetic situation of cells such as blood cells.Publication The intestinal microbiome and metabolome of dairy cows under challenging conditions(2022) Tröscher-Mußotter, Johanna; Seifert, JanaThe modern dairy cow is confronted with a multitude of stressors throughout live. Especially calving, transition, and microbial infections are strong challenges that can have long-lasting impacts on the cow’s health and performance. Yet, individuals can differ in their response towards these challenges, raising the question which characteristics in the dairy cow contribute to a more or less robust animal. Apart from genetics, the gut microbiome and the entailed metabolome is assumed to play an important role in buffering or promoting host stress. This is also due to the fact that the gut microbiome is strongly involved in the hosts energy metabolism and immune system. As dairy cows often show performance impairments during high energy demanding periods, it could be suggested that improving energy metabolism in these specific phases might reduce the negative phenotypic outcomes. This was tested using dietary L-carnitine, a metabolite inevitably necessary for energy metabolism. However, no supplement effects on the intestinal microbiome or metabolome have been found in the present work. Supplementation was continued throughout the complete trial. Calving functioned as an individual stimulus, and an intra-venous LPS injection induced a standardized inflammatory challenge, as a specific amount of LPS per kg of bodyweight was applied per cow. Supplemented animals were compared to a control group. In total, the animals were studied across 168 days and sampled extensively at several sites. The focus of this thesis was to analyze the bacterial consortia and metabolites of both, host and bacteria, in rumen, duodenum, and feces throughout the given period. This was to elucidate the metabolic reactions and bacterial shifts during the mentioned challenging periods and their response to the L-carnitine supplementation. First, the ruminal and duodenal fluid microbiome of eight double cannulated animals during the two respective challenges was analysed. Before calving and feed change, rumen and duodenal fluid bacterial consortia were significantly different, thereafter very alike. Strong microbial community shifts were observed throughout the complete trial irrespectively of the matrix. Both matrices varied in their metabolite patterns indicating functional variation among sites. Also, a strong increase of Bifidobacterium at three days after calving was observed in almost all animals pointing towards a strong biological purpose. This needs to be investigated in upcoming studies. The study could show increasing ketogenic activities in the animals after calving and proposes a possible protective host-microbial interaction, against a ruminal collapse induced by LPS challenge, here described as "microbial airbag". The second part included fecal samples of the same animals, which were analyzed for their bacterial consortia and targeted metabolites. Different dynamics and diversities of microbial communities amongst the individuals were observed, according to which animals could be grouped into three microbiome clusters. These showed in part fundamentally different metabolic, health, and performance parameters, indicating strong host-microbiome-metabolite interactions. The study demonstrated that microbiome clustering may contribute to identifying different metabo- and production types. Again, the study observed a strong increase of Bifidobacterium at three days after calving and even during the LPS challenge supporting the findings of the former study. This strengthens the hypothesis that also for the cow Bifidobacterium may have protective effects, as this genus is largely involved in health promoting activities. The power of this project lies in the massive sampling of different body sites in dairy cows across a very long period of time and finally, merging of the collected data. This, however, requires high computational efforts as numerous time points, matrices, animals, measurements, treatments, feeding regimen, and challenges resulted into a large bandwidth of parameters and metadata. Yet, it bears the potential to better elucidate and understand actions and reactions of the host, its microbiome and metabolism, as well as organ-axes in dairy cows and thereby gaining a more holistic picture of these complex animals. The aim of analyzing the host, its microbiome and metabolome throughout challenging periods resulted into the following main findings. Time, calving, and feed change remarkably change the microbial communities and to a lesser extent the metabolomes in all three matrices. Rumen and proximal duodenal fluid samples significantly differ in their metabolomes but not in their microbiome. In all matrices, an increase of Bifidobacterium is seen within three days after calving, which has to be further researched. Across the herd, three distinct microbiome clusters are found, which significantly differ in their production and health parameters.Publication The prediction of energy balance of dairy cows from animal, feed, and milk traits with special regard to milk fatty acids(2017) Becher, Vera; Rodehutscord, MarkusThe objective of the present study was to predict the energy balance (EB) of dairy cows from animal, feed, and milk traits. As the milk fatty acid (FA) profile is known to react to physiological conditions like an energy deficit, special regard was given to milk FA in order to identify new potential indicators for negative EB. Visiting six experimental stations in Germany, single milk samples were taken from dairy cows between their 6th and 133th day in milk to create a dataset covering a large spectrum of EB and a variety of practical diets. The milk composition was analyzed by mid-infrared spectrometry, and the milk FA profile via gas chromatography. Energy balance (MJ NEL/d), as response variable, was calculated by subtracting the cow’s energy requirements from energy intake. As candidate variables parity, day of lactation, dietary nutrient composition, milk yield, milk composition, and the milk FA profile were provided resulting in a pool of 62 potential predictors. The prediction of EB was performed in two different ways: first, an automated stepwise variable selection was performed with the whole variable pool (GLMs-N) and with FA only (GLMs-FA-N). As this method recently earned criticism, some other methods were also tested for a first variable selection: the regularized linear regression models Lasso, elastic net (ENET), adaptive Lasso (AdaLasso), and adaptive elastic net (ADAENET). As a machine learning method which also considers interactions and non-linear relationships random forests were also applied. The first variable selection was performed using a five-fold cross-validation which resulted in five models per selection method. All chosen effects were combined to one model (MODEL1) for each method, respectively. Following this, the individual effects of the MODEL1 were used for a forward selection based on the corrected Akaike Information Criterion (AICC) for further model reduction, resulting in MODEL2. Then, the non-significant effects were removed from the MODEL2, achieving the final MODEL3 for each method. The final models were validated using leave-one-out cross-validation. The models showed adequate correlations (r) between the predicted and the observed EB in leave-one-out cross-validation: although GLMs-FA-N had the lowest accuracy (r = 0.79), the result was still remarkable and showed how much information milk FA alone can provide. GLMs-N and AdaLasso performed best with r = 0.86 and 0.85 containing 21 and 18 predictors, respectively. However, other models like ADAENET achieved only slightly lower accuracy (r = 0.83) with only 6 predictors. The composition of the predictors was relatively similar in all models. All (except for GLMs-FA-N) contained days in milk, milk yield, C18:1c9, C15:0iso, and the ratio of omega-6 to omega-3 FA (n-6/n-3) as effects with the strongest impacts on the prediction. While milk yield, days in milk, and C18:1c9 mirrored physiologically obvious effects, the strong and positive impact of n-6/n-3 and C15:0iso was unexpected. The n-6/n-3 ratio might be physiologically connected to EB as might reflect the dietary forage-to-concentrate ratio which influences dietary energy content and thus EB. The importance of C15:0iso, a FA arising from microbial FA synthesis in the rumen, could not be explained satisfyingly. The nature of the potential physiological connections between EB and some FA like C15:0iso or n-6 or n-3 FA might require further research. The present study showed that it is possible to predict the cow’s EB from animal and milk traits with an adequate accuracy. As long as the diets have similar composition and not contain ingredients which strongly affect the milk FA profile, dietary effects have not to be taken into account. However, a practical application of the obtained models is not yet possible: First, as the dataset was relatively small (n = 248), it is not clear whether or not the models would perform adequately with independent datasets. Second, FA analysis by gas chromatography is very expensive. Third, even if gas chromatographic analysis were affordable for standard milk analysis, there are some highly variable, very low concentrated FA as predictors in the models, which might be prone to laboratory effects, and this could spoil the predictions. Although under criticism, automatic stepwise selection provided the best performing model and thus seems sufficient for practical issues like the one dealt with in the present study. However, the differences in accuracy between the applied methods were very small and as regularized linear regression methods, especially ENET and ADAENET, are supposed to deal better with highly correlated variables, it might be safer to use them with datasets containing highly correlated variables such as the one used in the present work.