Publications by authors named "Björn Kuhla"

49 Publications

Jejunal mucosa proteomics unravel metabolic adaptive processes to mild chronic heat stress in dairy cows.

Sci Rep 2021 Jun 14;11(1):12484. Epub 2021 Jun 14.

Institute of Nutritional Physiology "Oskar Kellner", Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany.

Climate change affects the duration and intensity of heat waves during summer months and jeopardizes animal health and welfare. High ambient temperatures cause heat stress in dairy cows resulting in a reduction of milk yield, feed intake, and alterations in gut barrier function. The objectives of this study were to investigate the mucosal amino acid, glucose and lactate metabolism, as well as the proteomic response of the small intestine in heat stressed (HS) Holstein dairy cows. Cows of the HS group (n = 5) were exposed for 4 days to 28 °C (THI = 76) in a climate chamber. Percentage decrease in daily ad libitum intake of HS cows was calculated to provide isocaloric energy intake to pair-fed control cows kept at 15 °C (THI = 60) for 4 days. The metabolite, mRNA and proteomic analyses revealed that HS induced incorrect protein folding, cellular destabilization, increased proteolytic degradation and protein kinase inhibitor activity, reduced glycolysis, and activation of NF-κB signaling, uronate cycling, pentose phosphate pathway, fatty acid and amino acid catabolism, mitochondrial respiration, ATPase activity and the antioxidative defence system. Our results highlight adaptive metabolic and immune mechanisms attempting to maintain the biological function in the small intestine of heat-stressed dairy cows.
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http://dx.doi.org/10.1038/s41598-021-92053-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8203643PMC
June 2021

Effects of 2 liquid feeding rates over the first 3 months of life on whole-body energy metabolism and energy use efficiency of dairy calves up to 5 months.

J Dairy Sci 2021 Jun 11. Epub 2021 Jun 11.

Institute of Nutritional Physiology "Oskar Kellner," Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 1, 18196 Dummerstorf, Germany. Electronic address:

Intensified milk replacer (MR) feeding in calves has nutritional long-term effects and is suggested to increase milk production later in life. However, the underlying mechanisms are not completely understood. The aim of our study was to investigate whether MR feeding intensity has long-term effects on energy metabolism and energy use efficiency of dairy calves. Newborn female Holstein calves (n = 28) were randomly assigned to 2 liquid feeding groups offered daily either 10% of body weight (BW) colostrum followed by 10% of BW MR (10%-MR) or 12% of BW colostrum followed by 20% of BW MR (20%-MR). Calves were housed individually. Weaning was completed by the end of wk 12. Hay and calf starter were fed from d 1 until the end of wk 14 and 16, respectively. A total mixed ration was fed from wk 11 onward, and the metabolizable energy intake (MEI) was determined daily. Energy metabolism of calves was measured in respiratory chambers before weaning in wk 6 and 9, and after weaning in wk 14 and 22. The MEI/BW was higher before weaning but lower during and shortly after weaning in 20%-MR calves. During the preweaning period, the 20%-MR animals had higher average daily gain, BW, back fat thickness and muscle diameter, but lower plasma β-hydroxybutyrate concentrations. The group difference in average daily gain ceased in wk 9, differences in back fat thickness and muscle diameter ceased after weaning, whereas difference in BW persisted until wk 23. The energy conversion ratio (BW gain/MEI) was not different before weaning, but was lower during and after weaning in 20%-MR calves. The higher MEI and BW in 20%-MR calves resulted in higher heat production (HP), as well as in higher carbohydrate oxidation (COX) and fat oxidation during the preweaning period. Gas exchange variables normalized to BW or MEI differed between groups only during preweaning. The energy balance was lower in 10%-MR calves in wk 6 and 9. The HP/BW and COX/BW were higher, whereas HP/MEI was lower in 20%-MR calves in wk 6. When normalized to BW and MEI, HP in wk 6 and 9, and COX in wk 9 was lower in 20%-MR calves. In conclusion, 20%-MR calves showed greater efficiency estimates preweaning, but this effect did not occur after weaning, suggesting that energy use efficiency does not persist until later stages in life.
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http://dx.doi.org/10.3168/jds.2021-20278DOI Listing
June 2021

Holstein dairy cows with high phosphorus utilization efficiency fed a low phosphorous diet secreted less phosphorus with urine but more with milk and feces.

Sci Total Environ 2021 Sep 17;788:147813. Epub 2021 May 17.

Institute of Nutritional Physiology 'Oskar Kellner', Research Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany. Electronic address:

The environmental pollution of phosphorus (P) from livestock farming is becoming increasingly problematic especially with regard to dwindling global P resources. Thus, a more sustainable handling of P resources, including improvements in P use efficiency and a reduction of P loss from farm animals, is necessary. Dairy cows may differ in milk P yield and P use efficiency despite receiving the same feed ration. The objective of this study was to elucidate inter-individual differences in P and closely linked nitrogen (N) excretions and the expression of P transport proteins in dairy cows with low and high P utilization efficiency. Twenty multiparous, late lactating German Holstein dairy cows were retrospectively assigned to either a high (HPeff; n = 10) or low (LPeff; n = 10) P utilization efficiency group. Cows were fed a diet low in P and crude protein (CP) content. During a 4-day balance study, feed intake, urine and fecal excretions, and milk yield were recorded to determine total P and N content in subsamples. Mammary gland, kidney and jejunal mucosa were sampled to analyze mRNA expressions of P transporters by real-time-PCR. A high milk P yield in HPeff cows strongly correlated with milk protein and milk N yield. HPeff cows excreted less urinary P, had a higher renal P reabsorption rate, and a higher renal sodium-P cotransporter 2 expression than LPeff cows. As HPeff cows channeled more P into milk, they mobilized more P from body reserves as indicated by their more negative P-balance. In addition, HPeff cows had higher fecal P excretion relative to ingested P, resulting in a lower apparent P digestibility. In conclusion, when fed a low P diet, HPeff cows channeled more endogenous P into milk and feces, which in the long-term, likely has adverse effects on animal health and the environment.
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http://dx.doi.org/10.1016/j.scitotenv.2021.147813DOI Listing
September 2021

Challenges and opportunities to capture dietary effects in on-farm greenhouse gas emissions models of ruminant systems.

Sci Total Environ 2021 May 8;769:144989. Epub 2021 Jan 8.

Instituto de Investigaciones Agropecuarias, INIA Remehue, Chile.

This paper reviews existing on-farm GHG accounting models for dairy cattle systems and their ability to capture the effect of dietary strategies in GHG abatement. The focus is on methane (CH) emissions from enteric and manure (animal excreta) sources and nitrous oxide (NO) emissions from animal excreta. We identified three generic modelling approaches, based on the degree to which models capture diet-related characteristics: from 'none' (Type 1) to 'some' by combining key diet parameters with emission factors (EF) (Type 2) to 'many' by using process-based modelling (Type 3). Most of the selected on-farm GHG models have adopted a Type 2 approach, but a few hybrid Type 2 / Type 3 approaches have been developed recently that combine empirical modelling (through the use of CH and/or NO emission factors; EF) and process-based modelling (mostly through rumen and whole tract fermentation and digestion). Empirical models comprising key dietary inputs (i.e., dry matter intake and organic matter digestibility) can predict CH and NO emissions with reasonable accuracy. However, the impact of GHG mitigation strategies often needs to be assessed in a more integrated way, and Type 1 and Type 2 models frequently lack the biological foundation to do this. Only Type 3 models represent underlying mechanisms such as ruminal and total-tract digestive processes and excreta composition that can capture dietary effects on GHG emissions in a more biological manner. Overall, the better a model can simulate rumen function, the greater the opportunity to include diet characteristics in addition to commonly used variables, and thus the greater the opportunity to capture dietary mitigation strategies. The value of capturing the effect of additional animal feed characteristics on the prediction of on-farm GHG emissions needs to be carefully balanced against gains in accuracy, the need for additional input and activity data, and the variability encountered on-farm.
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http://dx.doi.org/10.1016/j.scitotenv.2021.144989DOI Listing
May 2021

Improving robustness and accuracy of predicted daily methane emissions of dairy cows using milk mid-infrared spectra.

J Sci Food Agric 2021 Jun 20;101(8):3394-3403. Epub 2020 Dec 20.

AGROBIOCHEM Department and Research and Teaching Centre (TERRA), Gembloux Agro-Bio Tech, University of Liège, Gembloux, Belgium.

Background: A robust proxy for estimating methane (CH ) emissions of individual dairy cows would be valuable especially for selective breeding. This study aimed to improve the robustness and accuracy of prediction models that estimate daily CH emissions from milk Fourier transform mid-infrared (FT-MIR) spectra by (i) increasing the reference dataset and (ii) adjusting for routinely recorded phenotypic information. Prediction equations for CH were developed using a combined dataset including daily CH measurements (n = 1089; g d ) collected using the SF tracer technique (n = 513) and measurements using respiration chambers (RC, n = 576). Furthermore, in addition to the milk FT-MIR spectra, the variables of milk yield (MY) on the test day, parity (P) and breed (B) of cows were included in the regression analysis as explanatory variables.

Results: Models developed based on a combined RC and SF dataset predicted the expected pattern in CH values (in g d ) during a lactation cycle, namely an increase during the first weeks after calving followed by a gradual decrease until the end of lactation. The model including MY, P and B information provided the best prediction results (cross-validation statistics: R = 0.68 and standard error = 57 g CH d ).

Conclusions: The models developed accounted for more of the observed variability in CH emissions than previously developed models and thus were considered more robust. This approach is suitable for large-scale studies (e.g. animal genetic evaluation) where robustness is paramount for accurate predictions across a range of animal conditions. © 2020 Society of Chemical Industry.
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http://dx.doi.org/10.1002/jsfa.10969DOI Listing
June 2021

Differences between Holstein dairy cows in renal clearance rate of urea affect milk urea concentration and the relationship between milk urea and urinary nitrogen excretion.

Sci Total Environ 2021 Feb 27;755(Pt 2):143198. Epub 2020 Oct 27.

Institute of Nutritional Physiology 'Oskar Kellner', Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 1, 18196 Dummerstorf, Germany. Electronic address:

Urine and fecal excretions from cattle contribute to global nitrogen (N) emissions. The milk urea nitrogen (MUN) concentration in dairy cows is positively correlated with urinary urea N (UUN) emissions, and both decline with the reduction in crude protein intake. However, MUN concentration may differ between individual cows despite feeding the same ration. Thus, we hypothesized that due to differences in endogenous N utilization cows with high MUN concentration excrete more UUN than cows with a low MUN concentration. The objective of the present study was to elucidate N partitioning and urea metabolism in dairy cows with divergent MUN concentrations fed two planes of crude protein. Twenty Holstein dairy cows with high (HMU; n = 10) and low (LMU; n = 10) milk urea concentrations were fed two isocaloric diets with a low (LP) and normal (NP) crude protein level. Methane and ammonia emissions were recorded in respiration chambers. Feed intake, feces and urine excretions and milk yield were recorded for four days and subsamples were analyzed for total N and N-metabolites. A carbon-13 labeled urea bolus was administered intravenously followed by a series of plasma samplings. Total N and UUN excretions and ammonia emissions from excreta were lower on the LP diet, however, methane emissions, urinary N excretions and ammonia emissions were comparable between groups. Although plasma and salivary urea concentrations, urea pool size and urea turnover were higher, HMU cows had lower renal urea clearance rates. Additionally, HMU cows had lower renal clearance rates for creatinine, uric acid and creatine and excreted less uric acid (on the LP diet only) and creatine with urine. In conclusion, contrary to our hypothesis, HMU cows did not excrete more UUN than LMU cows. The lower urinary creatine excretion of HMU cows suggests that these animals have a lower environmental nitrogen footprint.
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http://dx.doi.org/10.1016/j.scitotenv.2020.143198DOI Listing
February 2021

The relationship between methane emission and daytime-dependent fecal archaeol concentration in lactating dairy cows fed two different diets.

Arch Anim Breed 2020 2;63(2):211-218. Epub 2020 Jul 2.

Institute of Animal Breeding and Husbandry, Kiel University, Kiel, 24098, Germany.

Archaeol is a cell membrane lipid of methanogenic archaea excreted in feces and is therefore a potential biomarker for individual methane emission (MEM). The aims of this study were to examine the potential of the fecal archaeol concentration (fArch) to be a proxy for MEM prediction in cows fed different diets and determine if the time of fecal collection affected the archaeol concentration. Thus, we investigated (i) the variation of the fArch concentration in spot samples of feces taken thrice within 8 h during respiration chamber measurements and (ii) the effect of two diets differing in nutrient composition and net energy content on the relationship between fArch and MEM in lactating cows. Two consecutive respiration trials with four primiparous and six multiparous lactating Holstein cows were performed. In the first trial (T1) at  d in milk (IM), a diet moderate in starch and fat content was fed for ad libitum intake, whereas in the second trial (T2) at  d IM, cows received a diet lower in starch and fat. Individual MEM (g d ) was measured for 24 h. Fecal samples were taken at 06:30, 10:00, and 14:30 LT and analyzed for fArch using Soxhlet lipid extraction and GC-MS. Cows produced less methane (364 g   d ) during T1 and had significantly lower fArch concentrations (37.1  g g dry matter; DM) compared to T2 (392 g   d and 47.6  g g  DM). A significant positive relationship between fArch ( g g fecal DM) and MEM, expressed on a dry matter intake (DMI) basis (g kg  DMI), was found ( , ). Among samples collected over the day, those collected at 10:00 LT provided the best coefficient of determination for MEM ( ). In conclusion, fArch offers some potential in serving as a proxy for innovative breeding schemes to lower enteric methane when fecal samples are taken at a certain time of the day, but more data on the sources of variation of the MEM   fArch ratios are required.
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http://dx.doi.org/10.5194/aab-63-211-2020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7397718PMC
July 2020

The effect of chronic, mild heat stress on metabolic changes of nutrition and adaptations in rumen papillae of lactating dairy cows.

J Dairy Sci 2020 Sep 26;103(9):8601-8614. Epub 2020 Jun 26.

Institute of Nutritional Physiology "Oskar Kellner," Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany. Electronic address:

Global warming and accompanying high ambient temperatures reduce feed intake of dairy cows and shift the blood flow from the core of the body to the periphery. As a result, hypoxia may occur in the digestive tract accompanied by disruption of the intestinal barrier, local endotoxemia and inflammation, and altered nutrient absorption. However, whether the barrier of the rumen, like the intestine, is affected by ambient heat has not been studied so far. Lactating Holstein dairy cows were subjected to heat stress at 28°C (temperature-humidity index = 76; n = 5) with ad libitum feed intake or to thermoneutral conditions at 15°C (temperature-humidity index = 60; n = 5) and pair-feeding to heat-stressed animals for a total of 4 d. Gas exchange and feed intake behavior were measured in a respiration chamber, and rumen epithelia were taken after slaughter. Heat stress significantly reduced meal size and whole-body fat oxidation but increased meal frequency and carbohydrate oxidation. The mRNA expression of toll-like receptor 4 (TLR4) and tight junction proteins and the phosphorylation of TLR4 downstream targets (interleukin-1 receptor-associated kinase 4, stress-activated protein kinase, p38 mitogen-activated protein kinase, and nuclear factor k-B) in the rumen epithelium were not affected by heat. The proteomics approach revealed increased expression of rumen epithelium proteins involved in the AMP-activated protein kinase (AMPK) and insulin signaling pathways in heat-stressed cows. Also, proteins involved in chaperone-mediated folding of proteins were upregulated, whereas those involved in antioxidant defense system were downregulated. Further, we found evidence for increased carbohydrate phosphorylation accompanied with an increased flux of carbohydrates through the hexosamine biosynthetic pathway, providing substrates for protein glycosylation. In conclusion, the mild heat stress did not induce barrier dysfunction or inflammatory responses in the rumen epithelium of dairy cows, probably because of adaptations in feed intake behavior and defense mechanisms at the tissue level.
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http://dx.doi.org/10.3168/jds.2020-18417DOI Listing
September 2020

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

J Dairy Sci 2020 Jul 14;103(7):6054-6069. Epub 2020 May 14.

Institute of Nutritional Physiology "Oskar Kellner," Leibniz Institute for Farm Animal Biology (FBN), 18196 Dummerstorf, Germany. Electronic address:

A growing need exists for the development of practical feeding strategies to mitigate methane (CH) emissions from cattle. Therefore, the objective of this study was to evaluate the influence of milk replacer feeding intensity (MFI) in calves on CH emission, rumen development, and performance. Twenty-eight female newborn Holstein calves were randomly assigned to 2 feeding groups, offered daily either 10% of the body weight (BW) in colostrum and subsequently 10% of the BW in milk replacer (MR; 10%-MR), or 12% of the BW in colostrum followed by 20% of the BW in MR (20%-MR). In wk 3, half of each feeding group was equipped with a permanent rumen cannula. Both groups were weaned at the end of wk 12. Hay and calf starter (mixture of pelleted grains) were offered from d 1 until wk 14 and 16, respectively. A total mixed ration was offered from wk 11 onward. Feed intake was measured daily and BW, anatomical measures, and rumen size weekly. Methane production and gastrointestinal passage rate were measured pre-weaning in wk 6 and 9 and post-weaning in wk 14 and 22, with additional estimation of organic matter digestibility. Rumen fluid, collected in wk 1, 2, 3, 6, 9, 14, 18, and 22, was analyzed for volatile fatty acid concentrations. Although the experimental period ended in wk 23, rumen volume of 17 calves was determined after slaughter in wk 34. Data was analyzed using ANOVA for the effects of feeding group, cannulation, and time, if applicable. Dry matter intake (DMI) of solid feed (SF) in 20%-MR animals was lower pre-weaning in wk 6 to 10 but mostly higher post-weaning. From wk 6 onward, anatomical measures and BW were greater in 20%-MR animals, and only the differences in body condition score gradually ceased post-weaning. Following the amount of SF intake, 10%-MR calves emitted more CH pre-weaning in wk 9, whereas post-weaning the 20%-MR group tended to have higher levels. Methane emission intensity (CH/BW) was lower pre-weaning in 20%-MR animals but was comparable to the 10%-MR group post-weaning. Methane yield (CH/DMI of SF) and estimated post-weaning organic matter digestibility were not affected by MFI. Rumen size normalized to heart girth was greater in 10%-MR calves from wk 5 to 10, but differences did not persist thereafter. In wk 34, rumen volume was higher in 20%-MR calves, but normalization to BW revealed no difference between feeding groups. In conclusion, high MFI reduces CH emission from calves pre-weaning, although this effect ceases post-weaning.
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http://dx.doi.org/10.3168/jds.2019-17875DOI Listing
July 2020

The use of milk Fourier transform mid-infrared spectra and milk yield to estimate heat production as a measure of efficiency of dairy cows.

J Anim Sci Biotechnol 2020 7;11:43. Epub 2020 May 7.

1Institute of Nutritional Physiology "Oskar Kellner," Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany.

Background: Transformation of feed energy ingested by ruminants into milk is accompanied by energy losses via fecal and urine excretions, fermentation gases and heat. Heat production may differ among dairy cows despite comparable milk yield and body weight. Therefore, heat production can be considered an indicator of metabolic efficiency and directly measured in respiration chambers. The latter is an accurate but time-consuming technique. In contrast, milk Fourier transform mid-infrared (FTIR) spectroscopy is an inexpensive high-throughput method and used to estimate different physiological traits in cows. Thus, this study aimed to develop a heat production prediction model using heat production measurements in respiration chambers, milk FTIR spectra and milk yield measurements from dairy cows.

Methods: Heat production was computed based on the animal's consumed oxygen, and produced carbon dioxide and methane in respiration chambers. Heat production data included 168 24-h-observations from 64 German Holstein and 20 dual-purpose Simmental cows. Animals were milked twice daily at 07:00 and 16:30 h in the respiration chambers. Milk yield was determined to predict heat production using a linear regression. Milk samples were collected from each milking and FTIR spectra were obtained with MilkoScan FT 6000. The average or milk yield-weighted average of the absorption spectra from the morning and afternoon milking were calculated to obtain a computed spectrum. A total of 288 wavenumbers per spectrum and the corresponding milk yield were used to develop the heat production model using partial least squares (PLS) regression.

Results: Measured heat production of studied animals ranged between 712 and 1470 kJ/kg BW. The coefficient of determination for the linear regression between milk yield and heat production was 0.46, whereas it was 0.23 for the FTIR spectra-based PLS model. The PLS prediction model using weighted average spectra and milk yield resulted in a cross-validation variance of 57% and a root mean square error of prediction of 86.5 kJ/kg BW. The ratio of performance to deviation (RPD) was 1.56.

Conclusion: The PLS model using weighted average FTIR spectra and milk yield has higher potential to predict heat production of dairy cows than models applying FTIR spectra or milk yield only.
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http://dx.doi.org/10.1186/s40104-020-00455-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7204237PMC
May 2020

Substitution of Dietary Sulfur Amino Acids by dl-2-Hydroxy-4-Methylthiobutyric Acid Reduces Fractional Glutathione Synthesis in Weaned Piglets.

J Nutr 2020 04;150(4):722-729

Institute of Nutritional Physiology "Oskar Kellner", Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.

Background: Cys is limiting for reduced glutathione (GSH) synthesis and can be synthesized from Met. We hypothesized that the dietary Met hydroxyl analogue dl-2-hydroxy-4-methylthiobutyric acid (dl-HMTBA) affects Cys and GSH metabolism and oxidative stress defense differently than Met.

Objective: The objective was to elucidate whether dl-HMTBA supplementation of a Met-deficient diet affects Cys flux, GSH fractional synthetic rate (FSR), and the basal oxidative stress level relative to Met supplementation in pigs.

Methods: Twenty-nine male German Landrace piglets aged 28 d were allocated to 3 dietary groups: a basal diet limiting in Met (69% of Met plus Cys requirement) supplemented with either 0.15% l-Met (LMET; n = 9), 0.15% dl-Met (DLMET; n = 11), or 0.17% dl-HMTBA (DLHMTBA; n = 9) on an equimolar basis. At age 54 d the pigs received a continuous infusion of [1-13C]-Cys to calculate Cys flux and Cys oxidation. After 3 d, GSH FSR was determined by [2,2-2H2]-glycine infusion, and RBC GSH and oxidized GSH concentrations were measured. At age 62 d the animals were killed to determine hepatic mRNA abundances of enzymes involved in GSH metabolism, GSH concentrations, and plasma oxidative stress defense markers.

Results: The Cys oxidation was 21-39% and Cys flux 5-15% higher in the fed relative to the feed-deprived state (P < 0.001). On average, GSH FSR was 49% lower (P < 0.01), and RBC GSH and total GSH concentrations were 12% and 9% lower, respectively, in DLHMTBA and DLMET relative to LMET pigs (P < 0.05). In the feed-deprived state, Gly flux, the GSH:oxidized glutathione (GSSG) ratio, RBC GSSG concentrations, plasma oxidative stress markers, and the hepatic GSH content did not differ between groups.

Conclusions: Although GSH FSR was higher in LMET compared with DLMET or DLHMTBA feed-deprived pigs, these differences were not reflected by lower oxidative stress markers and antioxidant defense enzymes in LMET pigs.
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http://dx.doi.org/10.1093/jn/nxz272DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7138682PMC
April 2020

Comparison of Methods to Measure Methane for Use in Genetic Evaluation of Dairy Cattle.

Animals (Basel) 2019 Oct 21;9(10). Epub 2019 Oct 21.

Agri-Food and Biosciences Institute, Hillsborough, Co. Down BT26 6DR, UK.

Partners in Expert Working Group WG2 of the COST Action METHAGENE have used several methods for measuring methane output by individual dairy cattle under various environmental conditions. Methods included respiration chambers, the sulphur hexafluoride (SF) tracer technique, breath sampling during milking or feeding, the GreenFeed system, and the laser methane detector. The aim of the current study was to review and compare the suitability of methods for large-scale measurements of methane output by individual animals, which may be combined with other databases for genetic evaluations. Accuracy, precision and correlation between methods were assessed. Accuracy and precision are important, but data from different sources can be weighted or adjusted when combined if they are suitably correlated with the 'true' value. All methods showed high correlations with respiration chambers. Comparisons among alternative methods generally had lower correlations than comparisons with respiration chambers, despite higher numbers of animals and in most cases simultaneous repeated measures per cow per method. Lower correlations could be due to increased variability and imprecision of alternative methods, or maybe different aspects of methane emission are captured using different methods. Results confirm that there is sufficient correlation between methods for measurements from all methods to be combined for international genetic studies and provide a much-needed framework for comparing genetic correlations between methods should these become available.
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http://dx.doi.org/10.3390/ani9100837DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6826463PMC
October 2019

Involvement of Plasma Endocannabinoids and the Hypothalamic Endocannabinoid System in Increasing Feed Intake after Parturition of Dairy Cows.

Neuroendocrinology 2020 29;110(3-4):246-257. Epub 2019 May 29.

Institute for Experimental Surgery, Rostock University Medical Center, Rostock, Germany.

The endocannabinoids (ECs) N-arachidonylethanolamide (anandamide; AEA) and 2-arachidonoylglycerol (2-AG) participate in the control of feed intake and energy metabolism. Most mammals increase their feed intake after parturition to cope with the increased energy and nutrient requirements for milk synthesis, thereby increasing their metabolic rate. Here we investigated in experiment 1 the regulation of plasma AEA and 2-AG concentrations during the transition from late pregnancy to early lactation in dairy cows, and analyzed in experiment 2 the expression of the EC system in the paraventricular nucleus (PVN) and the arcuate nucleus (ARC) of the hypothalamus of late and early lactating cows using immunohistochemistry. Cows in experiment 1 were retrospectively grouped based on peak plasma fatty acid concentrations to a high (H) or low (L) group. Feed intake was not different between groups before parturition, but was lower in H than L cows during early lactation. Plasma AEA and 2-AG concentrations increased 2.2- to 2.4-fold during early lactation, in which time plasma AEA concentrations rose faster in H cows than in L cows postpartum. Upregulation of N-acyl phosphatidylethanolamine-specific phospholipase D together with tending increased cannabinoid receptor 1 (CB1) expression, and downregulation of fatty acid amide hydrolase in early lactating cows suggested an increased PVN AEA tone. The abundance of CB1 in the ARC and diacylglycerol lipase-alpha was not different between late and early lactating cows, but PVN monoacylglycerol lipase expression was 30% higher in early lactating cows, indicating diminished PVN 2-AG concentrations. The results show a potential involvement of AEA in stimulating feed intake and of 2-AG in regulating energy metabolism of early lactating cows.
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http://dx.doi.org/10.1159/000501208DOI Listing
February 2021

Characterizing the metabotype and its persistency in lactating Holstein cows: An approach toward metabolic efficiency measures.

J Dairy Sci 2019 Jul 15;102(7):6559-6570. Epub 2019 May 15.

Institute of Nutritional Physiology "Oskar Kellner", Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany. Electronic address:

The variation in feed efficiency among dairy cows is due to differences in fermentation and digestion characteristics, but recent studies have suggested that various aspects of postabsorptive metabolic processes including heat production or the metabolizable energy for maintenance are more crucial. Thus, metabolic efficiency largely determines feed efficiency, but whether divergent feed efficient cows differ in O consumption and metabolic CO production, directly determining the metabolic rate has not been investigated. Therefore, the objective of the present study was to determine whether variation in ME intake (MEI), O consumption, and metabolic CO production account for the variation in metabolic efficiency of dairy cows and whether this effect persists across the lactation cycle. Eighteen cows with different German breeding value functional herd life were kept in freestalls with ad libitum access to a total mixed ration that was kept constant in composition throughout the first lactation. Cows were blood sampled and weighed at wk 5, 13, and 42 postpartum (pp) and transferred into respiration chambers. Animals were retrospectively clustered according to MEI, O consumption, and metabolic CO production, each normalized to metabolic body weight (mBW). Cluster analysis revealed 9 high metabolically efficient (high-Meff) and 9 low metabolically efficient cows. The high-Meff cows had greater MEI and feed conversion efficiency, produced less metabolic CO and methane, had a stronger negative energy balance, and tended to have a lower metabolic respiratory quotient. Further, high-Meff cows had lower residual MEI, less heat energy loss, and lower plasma glucose concentrations, but used a greater portion of body reserves instead of feed energy for milk synthesis, particularly at wk 5 and 13 pp. However, these group differences did not persist by wk 42 pp. Cow groups were not different in O consumption, milk yield, metabolizable energy for maintenance, or the efficiency of tissue utilization for milk synthesis, but high-Meff cows tended to have the lower German relative breeding value functional herd life, indicating a link between metabolic performance and productive lifespan. In conclusion, the use of a clustering approach involving MEI/mBW, O/mBW, and CO/mBW seems to be a promising method to differentiate cows with divergent metabolic efficiency but does not allow identifying an individual metabotype that persists across the whole lactation cycle.
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http://dx.doi.org/10.3168/jds.2019-16274DOI Listing
July 2019

Heat stress directly impairs gut integrity and recruits distinct immune cell populations into the bovine intestine.

Proc Natl Acad Sci U S A 2019 05 7;116(21):10333-10338. Epub 2019 May 7.

Institute of Genome Biology, FBN, 18196 Dummerstorf, Germany.

High ambient temperature has multiple potential effects on the organism such as hyperthermia, endotoxemia, and/or systemic inflammation. However, it is often difficult to discriminate between cause and consequence of phenotypic effects, such as the indirect influence of heat stress via reduced food intake. Lactating dairy cows are a particularly sensitive model to examine the effects of heat stress due to their intensive metabolic heat production and small surface:volume ratio. Results from this model show heat stress directly induced a so-far unknown infiltration of yet uncategorized cells into the mucosa and submucosa of the jejunum. Due to a pair-feeding design, we can exclude this effect being a consequence of the concurrent heat-induced reduction in feed intake. Isolation and characterization of the infiltrating cells using laser capture microdissection and RNA sequencing indicated a myeloic origin and macrophage-like phenotype. Furthermore, targeted transcriptome analyses provided evidence of activated immune- and phagocytosis-related pathways with LPS and cytokines as upstream regulators directly associated with heat stress. Finally, we obtained indication that heat stress may directly alter jejunal tight junction proteins suggesting an impaired intestinal barrier. The penetration of toxic and bacterial compounds during heat stress may have triggered a modulated immune repertoire and induced an antioxidative defense mechanism to maintain homeostasis between commensal bacteria and the jejunal immune system. Our bovine model indicates direct effects of heat stress on the jejunum of mammals already at moderately elevated ambient temperature. These results need to be considered when developing concepts to combat the negative consequences of heat stress.
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http://dx.doi.org/10.1073/pnas.1820130116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6535017PMC
May 2019

Substitution of Dietary Sulfur Amino Acids by DL-2-hydroxy-4-Methylthiobutyric Acid Increases Remethylation and Decreases Transsulfuration in Weaned Piglets.

J Nutr 2019 03;149(3):432-440

Institute of Nutritional Physiology, Dummerstorf, Germany.

Background: DL-2-hydroxy-4-methylthiobutyric acid (DL-HMTBA), an L-methionine (L-Met) hydroxyl analogue, has been suggested to be a dietary L-Met source. How dietary DL-HMTBA compared with L-Met affects whole-body L-Met kinetics in growing individuals is unknown.

Objectives: We determined to what extent DL-HMTBA supplementation of an L-Met-deficient diet affects whole-body L-Met and L-cysteine (L-Cys) kinetics, protein synthesis (PS), and the L-Met incorporation rate in liver protein (L-MetInc) compared with L-Met and DL-Met supplementation in a piglet model.

Methods: Forty-five, 28-d-old weaned piglets (male, German Landrace) were allocated to 4 dietary groups: L-Met-deficient diet [Control: 69% of recommended L-Met plus L-Cys supply; 0.22% standardized ileal digestible (SID) L-Met; 0.27% SID L-Cys; n = 12] and Control diet supplemented equimolarly to 100% of recommended intake with either L-Met (n = 12; LMET), DL-Met (n = 11; DLMET), or DL-HMTBA (n = 10; DLHMTBA). At 47 d of age, the piglets were infused with L-[1-13C; methyl-2H3]-Met and [3,3-2H2]-Cys to determine the kinetics and PS rates. Plasma amino acid (AA) concentrations, hepatic mRNA abundances of L-Met cycle and transsulfuration (TS) enzymes, and L-MetInc were measured.

Results: During feed deprivation, L-Met kinetics did not differ between groups, and were ≤3 times higher in the fed state (P < 0.01). Remethylation (RM) was 31% and 45% higher in DLHMTBA than in DLMET and Control pigs, respectively, and the RM:transmethylation (TM) ratio was 50% higher in DLHMTBA than in LMET (P < 0.05). Furthermore, TS and the TS:TM ratio were 32% lower in DLHMTBA than in LMET (P < 0.05). L-MetInc was 42% lower in DLMET and DLHMTBA than in L-Met-deficient Control pigs, whereas plasma AA and hepatic mRNA abundances were similar among DL-HMTBA-, L-Met-, and DL-Met-supplemented pigs.

Conclusions: In piglets, DL-HMTBA compared with L-Met and DL-Met supplementation increases RM and reduces the TS rate to conserve L-Met, but all 3 Met isomers support growth at a comparable rate.
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http://dx.doi.org/10.1093/jn/nxy296DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6398387PMC
March 2019

Methane prediction based on individual or groups of milk fatty acids for dairy cows fed rations with or without linseed.

J Dairy Sci 2019 Feb 26;102(2):1788-1802. Epub 2018 Dec 26.

Institute of Nutritional Physiology "Oskar Kellner," Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany; Nutritional Physiology and Animal Nutrition, Faculty of Agriculture and Environmental Sciences, University of Rostock, 18059 Rostock, Germany. Electronic address:

Milk fatty acids (MFA) are a proxy for the prediction of CH emission from cows, and prediction differs with diet. Our objectives were (1) to compare the effect of diets on the relation between MFA profile and measured CH production, (2) to predict CH production based on 6 data sets differing in the number and type of MFA, and (3) to test whether additional inclusion of energy-corrected milk (ECM) yield or dry matter intake (DMI) as explanatory variables improves predictions. Twenty dairy cows were used. Four diets were used based on corn silage (CS) or grass silage (GS) without (L0) or with linseed (LS) supplementation. Ten cows were fed CS-L0 and CS-LS and the other 10 cows were fed GS-L0 and GS-LS in random order. In feeding wk 5 of each diet, CH production (L/d) was measured in respiration chambers for 48 h and milk was analyzed for MFA concentrations by gas chromatography. Specific CH prediction equations were obtained for L0-, LS-, GS-, and CS-based diets and for all 4 diets collectively and validated by an internal cross-validation. Models were developed containing either 43 identified MFA or a reduced set of 7 groups of biochemically related MFA plus C16:0 and C18:0. The CS and LS diets reduced CH production compared with GS and L0 diets, respectively. Methane yield (L/kg of DMI) reduction by LS was higher with CS than GS diets. The concentrations of C18:1 trans and n-3 MFA differed among GS and CS diets. The LS diets resulted in a higher proportion of unsaturated MFA at the expense of saturated MFA. When using the data set of 43 individual MFA to predict CH production (L/d), the cross-validation coefficient of determination (R) ranged from 0.47 to 0.92. When using groups of MFA variables, the R ranged from 0.31 to 0.84. The fit parameters of the latter models were improved by inclusion of ECM or DMI, but not when added to the data set of 43 MFA for all diets pooled. Models based on GS diets always had a lower prediction potential (R = 0.31 to 0.71) compared with data from CS diets (R = 0.56 to 0.92). Models based on LS diets produced lower prediction with data sets with reduced MFA variables (R = 0.62 to 0.68) compared with L0 diets (R = 0.67 to 0.80). The MFA C18:1 cis-9 and C24:0 and the monounsaturated FA occurred most often in models. In conclusion, models with a reduced number of MFA variables and ECM or DMI are suitable for CH prediction, and CH prediction equations based on diets containing linseed resulted in lower prediction accuracy.
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http://dx.doi.org/10.3168/jds.2018-14911DOI Listing
February 2019

TECHNICAL NOTE: Development of a pressure sensor-based system for measuring rumination time in pre-weaned dairy calves.

J Anim Sci 2018 Nov;96(11):4483-4489

Institute of Nutritional Physiology, Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.

The pressure-based noseband sensor system (RWS: RumiWatch System; ITIN + HOCH GmbH Feeding Technology, Liestal, Switzerland) has recently been validated for the measurement of rumination time in mature cows. We aimed in this study at developing a similar pressure-based system for monitoring rumination in young dairy calves. To this end, a vegetable oil-filled silicon tube with a built-in pressure sensor (outer diameter 5.7 mm, length 38 cm) was attached to the noseband of a calf halter. In contrast to the RWS developed for mature cows, the accelerometer, the battery, the data logger, and the SD card of the RWS were integrated into 1 box to reduce the weight of the RWS to 0.35 kg. The box was attached to the halter so that it was located behind the right ear of the calf. Ten pre-weaned German Holstein calves (49-106 kg BW and 33-63 days of age) were equipped with the RWS. Calves were milk-fed thrice a day and offered hay and commercial starter for ad libitum intake. In parallel, animals were monitored by a video camera connected to a video recorder for 12 h. Two independent observers assessed the video records to obtain a reliable gold standard for the evaluation of the newly developed RWS. Data obtained by either RWS or visual video observation were processed as min rumination per h, yielding a total of 120 pairs of values (12 pairs per animal) for regression analysis. Assessment of 2 independent observers were highly correlated (r = 0.99). Results indicated relatively low random error between results obtained from the RWS (on y-axis) and video observations (on x-axis) (R2 = 0.82). However, the intercept of the regression line (y = 7.70 + 0.64 x) was significantly different from zero (P < 0.01) and the 95% confidence interval of the slope (0.79-0.94) did not include the value of 1. This translates to a significant systemic error resulting in overestimation of rumination time which is attributable to nutritive and nonnutritive oral activities that almost exclusively lasted for up to 10 min. Exclusion of false positive rumination signals lasting less than 10 or 5 consecutive min from the analysis reduced the random and systemic errors of the model (R2 = 0.86 and 0.93, respectively). We conclude that the newly developed RWS can be used to provide accurate measurement of rumination time in young calves. However, an extra programmed algorithm in the evaluation software is recommended to make the system more user-friendly for measurements on calves.
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http://dx.doi.org/10.1093/jas/sky337DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6247832PMC
November 2018

Feed-efficient pigs exhibit molecular patterns allowing a timely circulation of hormones and nutrients.

Physiol Genomics 2018 09 15;50(9):726-734. Epub 2018 Jun 15.

Institute of Genome Biology, Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.

Feed efficiency (FE) is a measure of the rate between feed intake and body weight gain and is subject to constant progress in pigs, based on extensive performance tests and analyses of physiological parameters. However, endocrine regulatory circuits that comprise the sensation and perception of intrinsic requirements and appropriate systemic responses have not yet been fully elucidated. It is hypothesized that the gut-brain axis, which is a network of hierarchical anterior regulatory tissues, contributes largely to variations in FE. Therefore, full-sib pigs with extreme residual feed intake values were assigned to experimental groups of high and low FE. Relevant hormones, minerals, and metabolites including fatty acid profiles were analyzed in serum to assess postprandial conditions. Transcriptome profiles were deduced from intestinal (duodenum, jejunum, ileum) and neuroendocrine tissues (hypothalamus). Serum analyses of feed-efficient animals showed an increased content of the incretin GIP, calcium, magnesium, β-hydroxybutyric acid, and fat compared with low-FE pigs. Complementary expression profiles in intestinal tissues indicate a modulated permeability and host-microbe interaction in FE-divergent pigs. Transcriptomic analyses of the hypothalamus showed that differences between the FE groups in appetite and satiety regulation are less pronounced. However, hypothalamic abundance of transcripts like ADCY7, LHCGR, and SLC2A7 and molecular signatures in local and systemic tissue sites indicate that increased allocation and circulation of energy equivalents, minerals, and hormones are promoted in feed-efficient animals. Overall, patterns of gastrointestinal hormones and gene expression profiles identified host-microbiota interaction, intestinal permeability, feed intake regulation, and energy expenditure as potential mechanisms affecting FE in pigs.
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http://dx.doi.org/10.1152/physiolgenomics.00021.2018DOI Listing
September 2018

Intravenous lipid infusion affects dry matter intake, methane yield, and rumen bacteria structure in late-lactating Holstein cows.

J Dairy Sci 2018 Jul 28;101(7):6032-6046. Epub 2018 Mar 28.

Institute of Nutritional Physiology "Oskar Kellner", Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany. Electronic address:

Increasing the dietary fat content of ruminant diets decreases methane (CH) production. This effect is caused by the toxic properties of fatty acids on rumen microbial populations, coating of feed particles diminishing the accessibility for microbes, and a reduction in dry matter intake (DMI). The latter effect is caused by postabsorptive long-chain fatty acids eliciting anorexic signaling; however, whether circulating long-chain fatty acids affect rumen CH production alike is unknown. To approach this question, 5 rumen-cannulated Holstein cows in late lactation received 2 jugular catheters and were kept in respiration chambers to measure CH production and DMI for 48 h. In a crossover design, cows were intravenously infused with a 20% lipid emulsion (LIPO) or 0.9% NaCl (CON). The LIPO cows received 2.1 kg of triglycerides/d [0.152 ± 0.007 g of triglycerides/(kg of BW × h)] consisting of 12.1% palmitic acid, 4.2% stearic acid, 31.1% oleic acid, and 52.7% linoleic acid. Blood and rumen fluid samples were taken hourly during the day. Results showed that LIPO compared with CON infusion increased plasma triglyceride as well as free fatty acid and serotonin concentrations but reduced the proportion of de novo synthesized milk fatty acids (sum of C6 to C16). Daily CH production and DMI were lower, whereas daily CH yield (CH/DMI) was greater in LIPO than CON cows, although CH yield decreased from d 1 to d 2 by 2 to 14% in LIPO-infused cows only. This effect was associated with a higher (acetate + butyrate)/propionate ratio, tending lower propionate concentrations between 24 and 34 h of infusion, reduced relative abundances of genera belonging to Succinivibrio, Ruminococcaceae, and Ruminiclostridium, and greater relative Bacteroidetes genus abundances in the rumen.
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http://dx.doi.org/10.3168/jds.2017-14101DOI Listing
July 2018

Variations in methane yield and microbial community profiles in the rumen of dairy cows as they pass through stages of first lactation.

J Dairy Sci 2018 Jun 15;101(6):5102-5114. Epub 2018 Mar 15.

School of Biology and Environmental Science & Earth Institute, University College Dublin, Dublin, D04 N2E5, Ireland. Electronic address:

Considerable interest exists both from an environmental and economic perspective in reducing methane emissions from agriculture. In ruminants, CH is produced by a complex community of microorganisms that is established in early life but can be influenced by external factors such as feed. Although CH emissions were thought to be constant once an animal reached maturity, recent studies have shown that CH yield significantly increases from early to late lactation in dairy cows. The aim of this study was to test the hypothesis that increases in CH yield over the lactation cycle are related to changes in rumen microbial community structure. Nine cows were monitored throughout their first lactation cycle. Methane and dry matter intake were measured to calculate CH per dry matter intake (CH yield) and ruminal fluid was collected during early, mid, and late lactation. A significant difference in bacterial and archaeal community structure during early and late lactation was observed. Furthermore, when ruminal short-chain fatty acid concentrations were measured, the ratio of acetate and butyrate to propionate was significantly higher in late lactation compared with early lactation. Propionate concentrations were higher in cows with low CH yield during late lactation, but no differences were observed in bacterial or archaeal community structures. Prevotella dominated the rumen of cows followed by Succinclasticum; Treponema, Fibrobacter, Ruminococcus, and Bifidobacterium were also in high abundance relative to other bacterial genera. In general, positive correlations were stronger between the most relatively abundant bacterial genera and acetate and butyrate concentrations in the cows with high CH and weaker between these genera and propionate concentration. This study indicates that increased CH yield in late lactation is reflected in significant changes in microbial community structure.
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http://dx.doi.org/10.3168/jds.2017-14200DOI Listing
June 2018

Hepatic thyroid signaling of heat-stressed late pregnant and early lactating cows.

J Endocrinol 2017 Aug 12;234(2):129-141. Epub 2017 May 12.

Institute of Nutritional Physiology 'Oskar Kellner'Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany

During the transition between late gestation and early lactation, dairy cows experience severe metabolic stress due to the high energy and nutrient requirements of the fetus and the mammary gland. Additional thermal stress that occurs with rising temperatures during the ongoing climate change has further adverse implications on energy intake, metabolism and welfare. The thyroid hormone (TH)-mediated cellular signaling has a pivotal role in regulation of body temperature, energy intake and metabolic adaptation to heat. To distinguish between energy intake and heat stress-related effects, Holstein cows were first kept at thermoneutrality at 15°C followed by exposure to heat stress (HS) at 28°C or pair-feeding (PF) at 15°C for 6 days, in late pregnancy and again in early lactation. Herein, we focused on hepatic metabolic changes associated with alterations in the hypothalamic-pituitary-thyroid axis in HS and PF animals. T and T levels dropped with HS or PF; however, in HS animals, this decline was more pronounced. Thyroid-stimulating hormone (TSH) levels remain unaffected, while plasma cholesterol concentrations were lower in HS than PF animals. Hepatic marker genes for TH action (, and ) decreased after HS and were lower compared to PF cows but only post-partum. Proteomics data revealed reduced hepatic amino acid catabolism ante-partum and a shift toward activated beta-oxidation and gluconeogenesis but declined oxidative stress defense post-partum. Thus, liver metabolism of HS and PF cows adapts differently to diminished energy intake both ante-partum and post-partum, and a different TH sensitivity is involved in the regulation of catabolic processes.
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http://dx.doi.org/10.1530/JOE-17-0066DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5516449PMC
August 2017

Exposure of Lactating Dairy Cows to Acute Pre-Ovulatory Heat Stress Affects Granulosa Cell-Specific Gene Expression Profiles in Dominant Follicles.

PLoS One 2016 17;11(8):e0160600. Epub 2016 Aug 17.

Institute of Nutritional Physiology, Leibniz Institute for Farm Animal Biology (FBN), 18196 Dummerstorf, Germany.

High environmental temperatures induce detrimental effects on various reproductive processes in cattle. According to the predicted global warming the number of days with unfavorable ambient temperatures will further increase. The objective of this study was to investigate effects of acute heat stress during the late pre-ovulatory phase on morphological, physiological and molecular parameters of dominant follicles in cycling cows during lactation. Eight German Holstein cows in established lactation were exposed to heat stress (28°C) or thermoneutral conditions (15°C) with pair-feeding for four days. After hormonal heat induction growth of the respective dominant follicles was monitored by ultrasonography for two days, then an ovulatory GnRH dose was given and follicular steroid hormones and granulosa cell-specific gene expression profiles were determined 23 hrs thereafter. The data showed that the pre-ovulatory growth of dominant follicles and the estradiol, but not the progesterone concentrations tended to be slightly affected. mRNA microarray and hierarchical cluster analysis revealed distinct expression profiles in granulosa cells derived from heat stressed compared to pair-fed animals. Among the 255 affected genes heatstress-, stress- or apoptosis associated genes were not present. But instead, we found up-regulation of genes essentially involved in G-protein coupled signaling pathways, extracellular matrix composition, and several members of the solute carrier family as well as up-regulation of FST encoding follistatin. In summary, the data of the present study show that acute pre-ovulatory heat stress can specifically alter gene expression profiles in granulosa cells, however without inducing stress related genes and pathways and suggestively can impair follicular growth due to affecting the activin-inhibin-follistatin system.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0160600PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4988698PMC
July 2017

Metabolic Response to Heat Stress in Late-Pregnant and Early Lactation Dairy Cows: Implications to Liver-Muscle Crosstalk.

PLoS One 2016 11;11(8):e0160912. Epub 2016 Aug 11.

Institute of Nutritional Physiology "Oskar Kellner", Leibnitz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.

Climate changes lead to rising temperatures during summer periods and dramatic economic losses in dairy production. Modern high-yielding dairy cows experience severe metabolic stress during the transition period between late gestation and early lactation to meet the high energy and nutrient requirements of the fetus or the mammary gland, and additional thermal stress during this time has adverse implications on metabolism and welfare. The mechanisms enabling metabolic adaptation to heat apart from the decline in feed intake and milk yield are not fully elucidated yet. To distinguish between feed intake and heat stress related effects, German Holstein dairy cows were first kept at thermoneutral conditions at 15°C followed by exposure to heat-stressed (HS) at 28°C or pair-feeding (PF) at 15°C for 6 days; in late-pregnancy and again in early lactation. Liver and muscle biopsies and plasma samples were taken to assess major metabolic pathway regulation using real-time PCR and Western Blot. The results indicate that during heat stress, late pregnant cows activate Cahill but reduce Cori cycling, prevent increase in skeletal muscle fatty acid oxidation, and utilize increased amounts of pyruvate for gluconeogenesis, without altering ureagenesis despite reduced plane of nutrition. These homeorhetic adaptations are employed to reduce endogenous heat production while diverting amino acids to the growing fetus. Metabolic adaptation to heat stress in early lactation involves increased long-chain fatty acid degradation in muscle peroxisomes, allowance for muscle glucose utilization but diminished hepatic use of amino acid-derived pyruvate for gluconeogenesis and reduced peroxisomal fatty acid oxidation and ATP production in liver of HS compared to PF cows in early lactation. Consequently, metabolic adaptation to heat stress and reduced feed intake differ between late pregnancy and early lactation of dairy cows to maintain energy supply for fetus development or milk production simultaneously reducing endogenous heat production.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0160912PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4981427PMC
August 2017

Methane emission, digestive characteristics and faecal archaeol in heifers fed diets based on silage from brown midrib maize as compared to conventional maize.

Arch Anim Nutr 2015 12;69(3):159-76. Epub 2015 May 12.

a Institute of Nutritional Physiology "Oskar Kellner", Leibniz Institute for Farm Animal Biology (FBN) , Dummerstorf , Germany.

The aim of the present experiment was to compare silage prepared from maize having a brown midrib (BMR) mutation with control (CTR) maize to identify their effects on enteric methane emission, digesta mean retention time (MRT), ruminal fermentation and digestibility. In addition, the utility of archaeol present in faecal samples was validated as a proxy for methane production. Seven German Holstein heifers were fed total mixed rations with a maize-silage proportion (either BMR or CTR) of 920 g/kg dry matter (DM) in a change-over design. Heifers were fed boluses with markers to measure MRT; faeces were collected for 7 days and rumen fluid was collected on the penultimate day. Methane emission was measured in respiration chambers on one day. Data were analysed by t-test and regression analysis. DM intake did not differ between the two diets. The apparent digestibility of DM and most nutrients was unaffected by diet type, but apparent digestibility of neutral and acid detergent-fibre was higher in those heifers fed BMR than in those fed CTR. Comparisons between diets revealed no difference in particle or solute MRT in the gastro-intestinal tract and the reticulorumen. Concentrations of short-chain fatty acid and ammonia in rumen fluid and its pH were not affected by silage type. Independent of the mode of expression [l/d, l/kg DM intake, l/kg digested organic matter], methane emissions were not affected by maize-silage type, but with BMR, there was a trend towards lower methane production per unit of digested neutral detergent fibre than there was with CTR silage. Results of the present study show that feeding heifers BMR silage does not increase methane emissions despite a higher fibre digestibility as compared to CTR silage. Therefore, it is assumed that improvements in animal productivity achieved by feeding BMR silage, as some studies have reported, can be obtained without extra environmental cost per unit of milk or meat. Neither faecal archaeol content [µg/g] nor daily amount excreted [mg/d] is suitable to predict methane production in absolute terms [l per day]. However, faecal archaeol content has a certain potential for predicting the methane yield [l per kg DM intake] of individual animals.
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http://dx.doi.org/10.1080/1745039X.2015.1043211DOI Listing
February 2016

Metabolic Heat Stress Adaption in Transition Cows: Differences in Macronutrient Oxidation between Late-Gestating and Early-Lactating German Holstein Dairy Cows.

PLoS One 2015 4;10(5):e0125264. Epub 2015 May 4.

Institute of Nutritional Physiology "Oskar Kellner", Leibniz Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.

High ambient temperatures have severe adverse effects on biological functions of high-yielding dairy cows. The metabolic adaption to heat stress was examined in 14 German Holsteins transition cows assigned to two groups, one heat-stressed (HS) and one pair-fed (PF) at the level of HS. After 6 days of thermoneutrality and ad libitum feeding (P1), cows were challenged for 6 days (P2) by heat stress (temperature humidity index (THI) = 76) or thermoneutral pair-feeding in climatic chambers 3 weeks ante partum and again 3 weeks post-partum. On the sixth day of each period P1 or P2, oxidative metabolism was analyzed for 24 hours in open circuit respiration chambers. Water and feed intake, vital parameters and milk yield were recorded. Daily blood samples were analyzed for glucose, β-hydroxybutyric acid, non-esterified fatty acids, urea, creatinine, methyl histidine, adrenaline and noradrenaline. In general, heat stress caused marked effects on water homeorhesis with impairments of renal function and a strong adrenergic response accompanied with a prevalence of carbohydrate oxidation over fat catabolism. Heat-stressed cows extensively degraded tissue protein as reflected by the increase of plasma urea, creatinine and methyl histidine concentrations. However, the acute metabolic heat stress response in dry cows differed from early-lactating cows as the prepartal adipose tissue was not refractory to lipolytic, adrenergic stimuli, and the rate of amino acid oxidation was lower than in the postpartal stage. Together with the lower endogenous metabolic heat load, metabolic adaption in dry cows is indicative for a higher heat tolerance and the prioritization of the nutritional requirements of the fast-growing near-term fetus. These findings indicate that the development of future nutritional strategies for attenuating impairments of health and performance due to ambient heat requires the consideration of the physiological stage of dairy cows.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0125264PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4418699PMC
February 2016

The control of short-term feed intake by metabolic oxidation in late-pregnant and early lactating dairy cows exposed to high ambient temperatures.

Physiol Behav 2015 Jun 31;145:64-70. Epub 2015 Mar 31.

Institute of Nutritional Physiology 'Oskar Kellner', Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany. Electronic address:

The objective of the present study was to integrate the dynamics of feed intake and metabolic oxidation in late pregnant and early lactating Holstein cows under heat stress conditions. On day 21 before parturition and again on day 20 after parturition, seven Holstein cows were kept for 7days at thermoneutral (TN) conditions (15°C; temperature-humidity-index (THI)=60) followed by a 7day heat stress (HS) period at 28°C (THI=76). On the last day of each temperature condition, gas exchange, feed intake and water intake were recorded every 6min in a respiration chamber. Pre- and post-partum cows responded to HS by decreasing feed intake. The reduction in feed intake in pre-partum cows was achieved through decreased meal size, meal duration, eating rate and daily eating time with no change in meal frequency, while post-partum cows kept under HS conditions showed variable responses in feeding behavior. In both pre- and post-partum cows exposed to heat stress, daily and resting metabolic heat production decreased while the periprandial respiratory quotient (RQ) increased. The prolonged time between meal and the postprandial minimum in fat oxidation and the postprandial RQ maximum, respectively, revealed that HS as compared to TN early-lactating cows have slower postprandial fat oxidation, longer feed digestion, and thereby showing a shift from fat to glucose utilization.
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http://dx.doi.org/10.1016/j.physbeh.2015.03.044DOI Listing
June 2015

Cerebrospinal fluid prohormone processing and neuropeptides stimulating feed intake of dairy cows during early lactation.

J Proteome Res 2015 Feb 14;14(2):823-8. Epub 2015 Jan 14.

Institute of Nutritional Physiology "Oskar Kellner", Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany.

After parturition, feed intake of dairy cows increases within the first weeks of lactation, but the molecular mechanisms stimulating or delaying the slope of increase are poorly understood. Some of the molecules controlling feed intake are neuropeptides that are synthesized as propeptides and subsequently processed before they bind to specific receptors in feeding centers of the brain. Cerebrospinal fluid surrounds most of the feed intake regulatory centers and contains numerous neuropeptides. In the present study, we used a proteomic approach to analyze the neuropeptide concentrations in cerebrospinal fluid taken from dairy cows between day -18 and -10, and between day +10 and +20 relative to parturition. We found 13 proteins which were only present in samples taken before parturition, 13 proteins which were only present in samples taken after parturition, and 25 proteins which were commonly present, before and after parturition. Among them, differences in pro-neuropeptide Y, proenkephalin-A, neuroendocrine convertase-2, neurosecretory protein VGF, chromogranin-A, and secretogranin-1 and -3 concentrations relative to parturition highlight propeptides and prohormone processings involved in the control of feed intake and energy homeostasis. Scaffold analysis further emphasized an increased tone of endogenous opioids associated with the postparturient increase of feed intake.
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http://dx.doi.org/10.1021/pr500872kDOI Listing
February 2015

Advanced glycation end products are mitogenic signals and trigger cell cycle reentry of neurons in Alzheimer's disease brain.

Neurobiol Aging 2015 Feb 13;36(2):753-61. Epub 2014 Oct 13.

Institute for Experimental Surgery, Medical School Rostock, University of Rostock, Rostock, Germany.

Neurons that reenter the cell cycle die rather than divide, a phenomenon that is associated with neurodegeneration in Alzheimer's disease (AD). Reexpression of cell-cycle related genes in differentiated neurons in AD might be rooted in aberrant mitogenic signaling. Because microglia and astroglia proliferate in the vicinity of amyloid plaques, it is likely that plaque components or factors secreted from plaque-activated glia induce neuronal mitogenic signaling. Advanced glycation end products (AGEs), protein-bound oxidation products of sugar, might be one of those mitogenic compounds. Cyclin D1 positive neurons are colocalized with AGEs or directly surrounded by extracellular AGE deposits in AD brain. However, a direct proof of DNA replication in these cells has been missing. Here, we report by using fluorescent in situ hybridization that consistent with the expression of cell cycle proteins, hyperploid neuronal cells are in colocalization with AGE staining in AD brains but not in nondemented controls. To complement human data, we used apolipoprotein E-deficient mice as model of neurodegeneration and showed that increased oxidative stress caused an intensified neuronal deposition of AGEs, being accompanied by an activation of the MAPK cascade via RAGE. This cascade, in turn, induced the expression of cyclin D1 and DNA replication. In addition, reduction of oxidative stress by application of α-lipoic acid decreased AGE accumulations, and this decrease was accompanied by a reduction in cell cycle reentry and a more euploid neuronal genome.
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http://dx.doi.org/10.1016/j.neurobiolaging.2014.09.025DOI Listing
February 2015