Gain and loss of subcutaneous and abdominal adipose tissue depot mass of German Holstein dairy cows with different body conditions during the transition period.

Subcutaneous adipose tissue (SCAT) and abdominal adipose tissue (AAT) depots are mobilized during the fresh cow period (FCP) and early lactation period (ELP) to counteract the negative energy balance (NEB). Earlier studies suggested that fat depots contribute differently to lipomobilization and may vary in functionality. Differences between the adipose depots might influence the development of metabolic disorders. Thus, the gain and loss of subcutaneous and abdominal adipose depot masses in Holstein cows with lower and higher body condition (mean body condition scores: 3.48 and 3.87, respectively) were compared in the period from d -42 to d 70 relative to parturition in this study. Animals of the 2 experimental groups represented adequately conditioned and overconditioned cows. Estimated depot mass (eDM) of SCAT, AAT, retroperitoneal, omental, and mesenteric adipose depots of 31 pluriparous German Holstein cows were determined via ultrasonography at d -42, 7, 28, and 70 relative to parturition. The cows were grouped according to the eDM of SCAT on d -42 [low body condition (LBC) group: n = 16, mean eDM 8.6 kg; high body condition (HBC) group: n = 15, mean eDM 15.6 kg]. Average daily change (prepartum gain and postpartum loss) in depot masses during dry period (DP; from d -42 to d 7), FCP (d 7 to d 28), and ELP (d 28 to d 70) were calculated and daily dry matter intake and lactation performance recorded. Cows of this study stored about 2 to 3 times more fat in AAT than in SCAT depots. After parturition, on average more adipose tissue mass was lost from the AAT than the SCAT depot (0.23 kg/d vs. 0.14 kg/d). Cows with high compared with low body condition had similar gains in AAT (0.33 kg/d) and SCAT (0.14 kg/d) masses during the DP but mobilized significantly more adipose tissue mass from both depots after calving (AAT, HBC vs. LBC: 0.30 vs. 0.17 kg/d; SCAT, HBC vs. LBC: 0.19 vs. 0.10 kg/d). Correlation analysis indicated a functional disparity between AAT and SCAT. In the case of AAT (R2 = 0.36), the higher the gain in adipose mass during DP, the higher the loss in FCP, but this was not the case for SCAT. During FCP, a greater NEB resulted in greater loss of mass from SCAT (R2 = 0.18). In turn, greater mobilization of SCAT mass led to a higher calculated feed efficiency (R2 = 0.18). However, AAT showed no such correlations. On the other hand, during ELP, loss of both SCAT and AAT mass correlated positively with feed efficiency (R2 = 0.35 and 0.33, respectively). The results indicate that feed efficiency may not be an adequate criterion for performance evaluation in cows during NEB. Greater knowledge of functional disparities between AAT and SCAT depots may improve our understanding of excessive lipomobilization and its consequences for metabolic health and performance of dairy cows during the transition period.

Modification of the equine gastrointestinal microbiota by Jerusalem artichoke meal supplementation.

The objective of this study was to investigate the impact of natural prebiotic active compounds on the microbial composition in different regions of the equine gastrointestinal tract. Twelve adult horses (body weight [bwt] 534 ± 64.5 kg; age 14 ± 7.5 years) were randomly divided into two feeding groups. Six horses received a basal diet consisting of 1.5 kg hay/100 kg bwt x d-1 and oat grains equal to 1.19 g starch/kg bwt x d-1, supplemented with Jerusalem artichoke meal providing prebiotic fructooligosaccharides + inulin in a quantity of 0.15 g/kg bwt x d-1. The remaining horses received a placebo added to the basal diet. The horses were fed for 21 d and euthanized at the end of the feeding period. Digesta samples from different parts of the gastrointestinal tract were taken, DNA extracted and the V1-V2 region of the 16S rRNA gene amplified. Supplementation with the prebiotic increased the relative abundance of Lactobacillus (P < 0.05), with a concurrent reduction of the relative abundance of Streptococcus mainly in the stomach (P < 0.05). In the hindgut, the supplemental prebiotic also increased the relative abundance of Lactobacillus but further reduced the relative abundance of fibrolytic bacteria, specifically the unclassified members of the families Lachnospiraceae (P < 0.05) and Ruminococcaceae. The relative abundance of the genus Ruminococcus increased solely in the caecum and colon transversum. Overall, the addition of the prebiotic significantly increased the diversity in nearly all parts of the gastrointestinal tract (P < 0.05). The feeding of this natural prebiotic compound to horses had an impact on the microbial community in the entire gastrointestinal tract. Furthermore, the effect on the bacterial community in the foregut (especially the stomach) was more pronounced in comparison to the effect in the hindgut. Therefore, the impact on stomach health should be carefully considered.

Does prebiotic feeding affect equine gastric health? A study on the effects of prebiotic-induced gastric butyric acid production on mucosal integrity of the equine stomach.

Fructo-oligosaccharides are commonly administered as prebiotics to horses in order to reduce the risk of disruption of microbial populations in the hindgut. Their microbial degradation to SCFA already begins in the stomach potentially resulting in increased gastric concentrations of SCFA such as butyric acid. The impact of butyric acid on the squamous mucosa is postulated to be detrimental, its effects on the glandular mucosa are yet unknown. Thus, the aim of this study was to determine the effects of butyric acid exposure on the functional integrity and morphology of the equine nonglandular and glandular gastric mucosa using butyric acid concentrations equivalent to the ones found in horses subjected to prebiotic fructo-oligosaccharides feeding. Gastric mucosal samples of healthy horses were exposed to butyric acid using the in vitro Ussing chamber technique. Electrophysiological parameters were continuously monitored, mucosal samples were blinded and histomorphological analysis was performed using a scoring system for assessment of histopathologic changes. Exposure to butyric acid resulted in pathohistomorphological changes in the glandular mucosa and in impairment of functional mucosal integrity in the squamous and the glandular mucosa as documented by significant changes in tissue conductances (Gt). Administration of fructo-oligosaccharides as a preventive prebiotic measure to horses should therefore be carefully considered, particularly in horses known to be at risk of developing EGUS.

Modulation of renal calcium and phosphate transporting proteins by dietary nitrogen and/or calcium in young goats.

In young goats, a reduction in dietary nitrogen (N) had an impact on mineral homeostasis although ruminants are able to recycle N effectively due to rumino-hepatic circulation. A solitary calcium (Ca) reduction stimulated calcitriol synthesis and Ca concentrations remained unchanged, whereas a dietary N reduction led to a decrease in calcitriol, which could not be prevented by a simultaneous reduction of N and Ca. In a previous study, it was shown that a reduced dietary N intake caused a decrease in intestinal Ca absorption due to a reduction of intestinal Ca transporting proteins. As no data on the potential role of the kidneys are available, it was the aim of the present study to evaluate whether an N- and/or Ca-reduced diet had an impact on renal Ca and phosphate (Pi) transporting protein expression in young goats. The animals were divided into 4 feeding groups, each receiving an adequate N and Ca supply, a reduced N supply, a reduced Ca supply, or a combined N and Ca reduction for 6 to 9 wk. The protein expression of the renal Ca channel transient receptor potential cation channel subfamily V member 5 (TRPV5) was diminished in N-reduced fed goats (P = 0.03), whereas in Ca restricted animals, the expression remained unaltered. The mRNA and protein expression of the Ca-binding protein calbindin-D28K (CaBPD28K) and the sodium-Ca exchanger 1 (NCX1) were significantly decreased due to the N-reduced feeding (mRNA, P = 0.003; P < 0.0001; protein, P = 0.002; P = 0.02), whereas dietary Ca reduction increased the CaBPD28K and NCX1 mRNA expression (P = 0.05; P = 0.01). The mRNA and protein expression of the parathyroid hormone receptor (PTHR) decreased due to the N-reduced feeding (P = 0.02; P = 0.03). These results confirm that a reduced dietary N intake led to decreased TRPV5, CaBPD28K, PTHR, and NCX1 expression levels, contributing to low levels of calcitriol and plasma Ca. In contrast to this, sodium-phosphate cotransporter type IIa expression and plasma Pi concentration were increased during dietary N reduction, thus indicating that Pi homeostasis is modulated in a calcitriol-independent manner. In conclusion, the modulation of Ca transporting proteins expression in the kidney is not able to prevent changes in mineral homeostasis in young goats receiving an N-reduced diet.

Dietary nitrogen and calcium modulate CYP27B1 expression in young goats.

In livestock, feeding a reduced nitrogen (N) diet is favored for economic and ecological reasons. Ruminants cope more easily with a reduced N diet than monogastric species. However, changes in mineral homeostasis such as a reduction in 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) concentrations, calcium (Ca), and IGF1 levels were observed in goats kept on a reduced N diet. The decrease in 1,25-(OH)2D3 occurred even during a simultaneous reduction in dietary N and Ca, whereas a solitary Ca reduction stimulated 1,25-(OH)2D3 synthesis. The aim of the present study was to examine the effects of N- and/or Ca-reduced diets on the expression of 24-hydroxylase (CYP24A1), 1-alpha-hydroxylase (CYP27B1), vitamin D receptor (VDR), retinoid X receptor alpha (RXRα), IGF1 receptor (IGF1R), Klotho, and fibroblast growth factor receptor 1c (FGFR1c) in kidneys of young goats. Four groups were kept on a control diet, an N-reduced diet, a Ca-reduced diet or an N- and a Ca-reduced diet. Renal expression of CYP24A1 was not affected, whereas CYP27B1 expression was significantly diminished in the N-reduced diet fed goats (P < 0.05) and significantly elevated with the Ca reduction (P < 0.001). The VDR expression was not modified, whereas RXRα (P < 0.05) and Klotho expression (P < 0.001) were stimulated during Ca reduction. The IGF1R (P < 0.05) and FGFR1c (P < 0.05) expression were enhanced with the N reduction. From these data, it can be concluded that the downregulation of renal CYP27B1 expression observed with dietary N reduction is probably mediated by a complex interaction between the somatotropic axis and the Klotho/FGF signaling pathway in young goats.

Interrelations between the rumen microbiota and production, behavioral, rumen fermentation, metabolic, and immunological attributes of dairy cows.

Different studies have shown a strong correlation between the rumen microbiome and a range of production traits (e.g., feed efficiency, milk yield and components) in dairy cows. Underlying dynamics concerning cause and effect are, however, still widely unknown and warrant further investigation. The aim of the current study was to describe possible functional interrelations and pathways using a large set of variables describing the production, the metabolic and immunological state, as well as the rumen microbiome and fermentation characteristics of dairy cows in early lactation (n = 36, 56 ± 3 d in milk). It was further hypothesized that the feed intake-associated behavior may influence the ruminal fermentation pattern, and a set of variables describing these individual animal attributes was included. Principal component analysis as well as Spearman's rank correlations were conducted including a total of 265 variables. The attained plots describe several well-known associations between metabolic, immunological, and production traits. Main drivers of variance within the data set included milk production and efficiency as well as rumen fermentation and microbiome diversity attributes, whereas behavioral, metabolic, and immunological variables did not exhibit any strong interrelations with the other variables. The previously well-documented strong correlation of production traits with distinct prokaryote groups was confirmed. This mainly included a negative correlation of operational taxonomic units ascribed to the Prevotella genus with milk and fat yield and feed efficiency. A central role of the animals' feed intake behavior in this context could not be affirmed. Furthermore, different methodological and interpretability aspects concerning the microbiome analysis by 16S rRNA gene sequencing, such as the discrepancy between taxonomic classification and functional communality, as well as the comparability with other studies, are discussed. We concluded that, to further investigate the driving force that causes the difference between efficient and inefficient animals, studies including more sophisticated methods to describe phenotypical traits of the host (e.g., rumen physiology, metabolic and genetic aspects) as well as the rumen microbiome (e.g., metagenome, metatranscriptome, metaproteome, and metabolome analysis) are needed.

Induction of a transient acidosis in the rumen simulation technique.

Feeding high concentrate diets to cattle results in an enhanced production of short-chain fatty acids by the micro-organisms in the rumen. Excessive fermentation might result in subclinical or clinical rumen acidosis, characterized by low pH, alterations in the microbial community and lactate production. Here, we provide an in vitro model of a severe rumen acidosis. A transient acidosis was induced in the rumen simulation technique by lowering bicarbonate, dihydrogen phosphate and hydrogen phosphate concentrations in the artificial saliva while providing a concentrate-to-forage ratio of 70:30. The experiment consisted of an equilibration period of 7 days, a first control period of 5 days, the acidosis period of 5 days and a second control period of 5 days. During acidosis induction, pH decreased stepwise until it ranged below 5.0 at the last day of acidosis (day 17). This was accompanied by an increase in lactate production reaching 11.3 mm at day 17. The daily production of acetate, propionate and butyrate was reduced at the end of the acidosis period. Gas production (methane and carbon dioxide) and NH3 -N concentration reached a minimum 2 days after terminating the acidosis challenge. While the initial pH was already restored 1 day after acidosis, alterations in the mentioned fermentation parameters lasted longer. However, by the end of the experiment, all parameters had recovered. An acidosis-induced alteration in the microbial community of bacteria and archaea was revealed by single-strand conformation polymorphism. For bacteria, the pre-acidotic community could be re-established within 5 days, however, not for archaea. This study provides an in vitro model for a transient rumen acidosis including biochemical and microbial changes, which might be used for testing feeding strategies or feed additives influencing rumen acidosis.

Relationships between body temperatures and inflammation indicators under physiological and pathophysiological conditions in pigs exposed to systemic lipopolysaccharide and dietary deoxynivalenol.

We studied the constancy of the relationship between rectal and intraabdominal temperature as well as their linkage to inflammatory markers (leucocyte counts, kynurenine-to-tryptophan ratio (Kyn-Trp ratio), tumour necrosis factor alpha (TNF-α) in healthy and in pigs exposed to lipopolysaccharide (LPS) and/or deoxynivalenol (DON). Barrows (n = 44) were fed 4 weeks either a DON-contaminated (4.59 mg DON/kg feed) or a control (CON) diet and equipped with an intraabdominal temperature logger and a multicatheter system (V.portae hepatis, V.lienalis, Vv.jugulares) facilitating infusion of 0.9% NaCl (CON) or LPS (7.5 µg/kg BW) and simultaneous blood sampling. Body temperatures were measured and blood samples taken every 15 min for leucocyte counts, TNF-α and Kyn-Trp ratio. Combination of diet and infusion created six groups: CON_CONjug .-CONpor. , CON_CONjug. -LPSpor. , CON_LPSjug. -CONpor. , DON_CONjug. -CONpor. , DON_CONjug. -LPSpor. , DON_LPSjug. -CONpor. . The relationship between both temperatures was not uniform for all conditions. Linear regression revealed that an intraabdominal increase per 1°C increase in rectal temperature was ~25% higher in all LPS-infused pigs compared to NaCl-infusion, albeit diet and site of LPS infusion modified the magnitude of this difference. Inflammatory markers were only strongly present under LPS influence and showed a significant relationship with body temperatures. For example, leucocyte counts in clinically inconspicuous animals were only significantly correlated to core temperature in DON-fed pigs, but in all LPS-infused groups, irrespective of diet and temperature method. In conclusion, the gradient between body core and rectal temperature is constant in clinically inconspicuous pigs, but not under various pathophysiological conditions. In the latter, measurement of inflammatory markers seems to be a useful completion.

Identification of hypoglycin A binding adsorbents as potential preventive measures in co-grazers of atypical myopathy affected horses.

BACKGROUND: Intestinal absorption of hypoglycin A (HGA) and its metabolism are considered major prerequisites for atypical myopathy (AM). The increasing incidence and the high mortality rate of AM urgently necessitate new therapeutic and/or preventative approaches. OBJECTIVES: To identify a substance for oral administration capable of binding HGA in the intestinal lumen and effectively reducing the intestinal absorption of the toxin. STUDY DESIGN: Experimental in vitro study. METHODS: Substances commonly used in equine practice (activated charcoal composition, di-tri-octahedral smectite, mineral oil and activated charcoal) were tested for their binding capacity for HGA using an in vitro incubation method. The substance most effective in binding HGA was subsequently tested for its potential to reduce intestinal HGA absorption. Jejunal tissues of 6 horses were incubated in Ussing chambers to determine mucosal uptake, tissue accumulation, and serosal release of HGA in the presence and absence of the target substance. Potential intestinal metabolism in methylenecyclopropyl acetic acid (MCPA)-conjugates was investigated by analysing their concentrations in samples from the Ussing chambers. RESULTS: Activated charcoal composition and activated charcoal were identified as potent HGA binding substances with dose and pH dependent binding capacity. There was no evidence of intestinal HGA metabolism. MAIN LIMITATIONS: Binding capacity of adsorbents was tested in vitro using aqueous solutions, and in vivo factors such as transit time and composition of intestinal content, may affect adsorption capacity after oral administration. CONCLUSIONS: For the first time, this study identifies substances capable of reducing HGA intestinal absorption. This might have major implications as a preventive measure in cograzers of AM affected horses but also in horses at an early stage of intoxication.

Influence of fumaric acid on ruminal parameters and organ weights of growing bulls fed with grass or maize silage.

The influence of the potential methane reducer, fumaric acid (FA), on ruminal parameters, the rumen wall and organ weights was investigated in a long-term study with growing bulls. In all, 20 bulls were fed with maize or grass silage as roughage, and with concentrate with or without 300 g FA per animal and day during the whole fattening period. After slaughtering, the organs were weighed and blood serum was analysed for glucose, ß-hydroxybutyric acid (BHB) and non-esterified fatty acid concentration. The ruminal fluid was analysed for short-chain fatty acids, ammonia-N and the microbial community via single strand conformation polymorphism analysis. The rumen wall was examined histopathologically and results were graded as 'no visible lesions', 'few inflammatory infiltrates', 'some inflammatory infiltrates' or 'several inflammatory infiltrates'. In addition, the dimensions of the rumen villi were measured. The FA supplementation decreased the serum BHB concentration and the butyric acid concentration in the ruminal fluid. The microbial community in the ruminal fluid was not influenced by FA. An interaction between FA and silage type was observed for the inflammation centres counted in the villous area of rumen papillae. This interaction was also observed in the length and surface of the rumen villi. Rumen villi results show that the influence of FA depends on the roughage used in the diet.

Differential effects of monensin and a blend of essential oils on rumen microbiota composition of transition dairy cows.

In response to oral application, monensin alters the rumen microbiota, increasing ruminal propionate production and energy availability in the animal. Data from different studies indicate that the susceptibility of rumen bacteria to monensin is mainly cell-wall dependent but tracing its activity to specific microbial groups has been challenging. Several studies have shown a similar effect for essential oils but results are inconsistent. To investigate the influence of monensin and a blend of essential oils (BEO, containing thymol, guaiacol, eugenol, vanillin, salicylaldehyde, and limonene) on the rumen microbiome, rumen liquid samples were collected orally on d 56 postpartum from cows that had either received a monensin controlled-release capsule 3 wk antepartum, a diet containing a BEO from 3 wk antepartum onward, or a control diet (n = 12). The samples were analyzed for pH, volatile fatty acid, ammonia, and lipopolysaccharide concentrations and protozoal counts. A 16S rRNA gene fingerprinting analysis (PCR-single-strand conformation polymorphism) and sequencing revealed that the BEO treatment had no effect on the rumen microbiota, whereas monensin decreased bacterial diversity. Twenty-three bacterial species-level operational taxonomic units were identified for which monensin caused a significant decrease in their relative abundance, all belonging to the phyla Bacteroidetes (uncultured BS11 gut group and BS9 gut group) and Firmicutes (Lachnospiraceae, Ruminococcaceae, and Erysipelotrichaceae). Ten bacterial operational taxonomic units belonging to the phyla Actinobacteria (Coriobacteriaceae), Bacteroidetes (Prevotella), Cyanobacteria (SHA-109), and Firmicutes (Lachnospiraceae and Ruminococcaceae) increased in relative abundance due to the monensin treatment. These results confirm the hypothesis that varying effects depending on cell-wall constitution and thickness might apply for monensin sensitivity rather than a clear-cut difference between gram-negative and gram-positive bacteria. No effect of monensin on the archaea population was observed, confirming the assumption that reported inhibition of methanogenesis is most likely caused through a decrease in substrate availability, rather than by a direct effect on the methanogens. The data support the hypothesis that the observed increase in ruminal molar propionate proportions due to monensin may be caused by a decrease in abundance of non-producers and moderate producers of propionate and an increase in abundance of succinate and propionate producers.

Segmental diversity of phosphate transport along the intestinal axis in horses.

For horses, distinct differences in intestinal phosphate transport have been postulated to account for the unique features of hind gut fermentation compared to other monogastric animals and ruminants. So far published data on mechanisms and underlying transport proteins involved in intestinal phosphate transport in the horse are still missing. Therefore we investigated intestinal phosphate transport in horses at both functional and molecular levels. Segmental diversity of intestinal phosphate transport along the intestinal axis was documented using the Ussing chamber technique. A transcellular phosphate secretion in the jejunum was confirmed. Furthermore, 2 sodium-dependent phosphate cotransporters, NaPiIIb and PiT1, were first detected in the equine intestine at mRNA level with PiT1 being expressed in both the small and large intestine, and NaPiIIb being solely expressed in the large intestine. In the colon, unidirectional net flux rates of phosphate were significantly greater compared to flux rates in other segments ( < 0.005) suggesting the colon as a major site for phosphate absorption in horses. Phosphate transport in the colon was mainly transcellular and mediated by a sodium-gradient as documented by Ussing chamber experiments and uptake of phosphate into colonic brush border membrane vesicles. In summary, the present study demonstrated mechanisms and transporters of intestinal phosphate transport in equine intestinal tissues with distinct differences between intestinal segments providing a new basis for a better understanding of intestinal phosphate transport in horses.

Trans- and paracellular calcium transport along the small and large intestine in horses.

Intestinal calcium absorption plays a key role in the maintenance of calcium homeostasis and may either occur by paracellular or transcellular mechanisms. The horse has some unique peculiarities in calcium homeostasis compared to other species including a high absorptive capacity for calcium in the intestine, high plasma calcium concentrations, high renal excretion, and low plasma concentrations of vitamin D metabolites. So far, knowledge about the underlying mechanisms and the regulation of intestinal calcium absorption is still limited concerning this species. Several studies have documented that intestinal calcium transport in horses is not as dependent on vitamin D as in other species. However, published data on other potential regulatory mechanisms are still lacking. In the present study, paracellular and transcellular transport mechanisms for intestinal calcium transport along the intestinal axis were identified in horses using the Ussing chamber technique. Furthermore, the expression of respective transport proteins including transient receptor potential vanilloid member 6, calbindin-D9k and calcium ATPase type 1 in line with the determined calcium flux rates was documented. In respect to regulation of transepithelial calcium transport, novel regulatory proteins for maintaining calcium homeostasis such as B-box and SPRY-domain containing protein and calmodulin were investigated for the first time in equine intestinal tissues in this study. This provides the basis for a new approach for a better understanding of equine calcium homeostasis regulation.

Haematological and febrile response to Escherichia coli lipopolysaccharide in 12-week-old cockerels of genetically diverse layer lines fed diets with increasing L-arginine levels.

Due to its decisive function in the avian metabolic, endocrine and immune system L-arginine (Arg) is dietary indispensable for chickens. In 12-week-old cockerels of two high- and two low-performing purebred layer lines, the effects of increasing dietary Arg on the haematological and febrile response were studied over 48 h after single lipopolysaccharide (LPS) injection. The offered diets contained Arg equivalent to 70%, 100% and 200% of recommended supply. Pathophysiological alterations in weight gain, feed intake, body temperature and differential blood count were examined in comparison with their physiological initial values. Within the first 24 h after LPS injection, cockerels reduced feed intake and lost body weight subsequently. Thereby, low-performing genotypes lost body weight to a lesser extent than high-performing ones. The loss of body weight was further intensified by deficient dietary Arg. Within the following 24 h, cockerels recovered by improving feed intake and weight gain. Furthermore, LPS induced genotype-specific fever response: both brown genotypes showed initial hypothermia followed by longer lasting moderate hyperthermia, whereas the white genotypes exhibited biphasic hyperthermia. Fever response was accompanied by significant changes in differential blood counts. Characterized by lymphopenia and heterophilia, a severe leucopenia was observed from 4 to 8 h after LPS injection and replaced by a marked leucocytosis with longer lasting monocytosis up to 48 h after LPS injection. Under given pathophysiological conditions, deficiently Arg-supplied cockerels showed higher total leucocyte counts than adequately and excessively Arg-supplied cockerels. However, deficient and surplus dietary Arg tended to cause higher ratios between heterophils and lymphocytes. To conclude, present results confirmed that LPS induced numerous immunological changes in 12-week-old cockerels and emphasized that chicken's genotype is a source of variation to be considered for immunological studies. Deficient dietary Arg intensified acute changes in differential blood counts and weight gain during LPS-induced inflammation.

Investigations on the possible impact of a glyphosate-containing herbicide on ruminal metabolism and bacteria in vitro by means of the 'Rumen Simulation Technique'.

AIMS: This study was performed in a well-established in vitro model to investigate whether the application of a glyphosate-containing herbicide might affect the bacterial communities and some biochemical parameters in a cow's rumen. METHODS AND RESULTS: The test item was applied in two concentrations (high and low) for 5 days. In a second trial, fermentation vessels were inoculated with Clostridium sporogenes before the high dose was applied. Effluents were analysed by biochemical, microbiological and genetic methods. A marginal increase in short-chain fatty acid production and a reduction in NH3 -N were observed. There were minor and rather equivocal changes in the composition of ruminal bacteria but no indications of a shift towards a more frequent abundance of pathogenic Clostridia species. Clostridium sporogenes counts declined consistently. CONCLUSIONS: No adverse effects of the herbicide on ruminal metabolism or composition of the bacterial communities could be detected. In particular, there was no evidence of a suspected stimulation of Clostridia growth. SIGNIFICANCE AND IMPACT OF THE STUDY: Antibiotic activity of glyphosate resulting in microbial imbalances has been postulated. In this exploratory study, however, intraruminal application of concentrations reflecting potential exposure of dairy cows or beef cattle did not exhibit significant effects on bacterial communities in a complex in vitro system. The low number of replicates (n = 3/dose) may leave some uncertainty.

Excretion pathways and ruminal disappearance of glyphosate and its degradation product aminomethylphosphonic acid in dairy cows.

From 6 balance experiments with total collection of feces and urine, samples were obtained to investigate the excretion pathways of glyphosate (GLY) in lactating dairy cows. Each experiment lasted for 26d. The first 21d served for adaptation to the diet, and during the remaining 5d collection of total feces and urine was conducted. Dry matter intake and milk yield were recorded daily and milk and feed samples were taken during the sampling periods. In 2 of the 6 experiments, at the sampling period for feces and urine, duodenal contents were collected for 5d. Cows were equipped with cannulas at the dorsal sac of the rumen and the proximal duodenum. Duodenal contents were collected every 2h over 5 consecutive days. The daily duodenal dry matter flow was measured by using chromium oxide as a volume marker. All samples (feed, feces, urine, milk and duodenal contents were analyzed for GLY and aminomethylphosphonic acid (AMPA). Overall, across the 6 experiments (n=32) the range of GLY intake was 0.08 to 6.67mg/d. The main proportion (61±11%; ±SD) of consumed GLY was excreted with feces; whereas excretion by urine was 8±3% of GLY intake. Elimination via milk was negligible. The GLY concentrations above the limit of quantification were not detected in any of the milk samples. A potential ruminal degradation of GLY to AMPA was derived from daily duodenal GLY flow. The apparent ruminal disappearance of GLY intake was 36 and 6%. In conclusion, the results of the present study indicate that the gastrointestinal absorption of GLY is of minor importance and fecal excretion represents the major excretion pathway. A degradation of GLY to AMPA by rumen microbes or a possible retention in the body has to be taken into account.

The effects of a ration change from a total mixed ration to pasture on rumen fermentation, volatile fatty acid absorption characteristics, and morphology of dairy cows.

To investigate the effect of the change from a concentrate and silage-based ration (total mixed ration, TMR) to a pasture-based ration, a 10-wk trial (wk 1-10) was performed, including 10 rumen- and duodenum-fistulated German Holstein dairy cows (182±24 d in milk, 23.5±3.5kg of milk/d; mean ± standard deviation). The cows were divided in either a pasture group (PG, n=5) or a confinement group (CG, n=5). The CG stayed on a TMR-based ration (35% corn silage, 35% grass silage, 30% concentrate; dry matter basis), whereas the PG was gradually transitioned from a TMR to a pasture-based ration (wk 1: TMR only; wk 2: 3 h/d on pasture wk 3 and 4: 12 h/d on pasture wk 5-10: pasture only). Ruminal pH, volatile fatty acids (VFA), NH3-N, and lipopolysaccharide (LPS) concentrations were measured in rumen fluid samples collected medially and ventrally on a weekly basis. Ruminal pH was continuously recorded during 1 to 4 consecutive days each week using ruminal pH measuring devices. In wk 1, 5, and 10, rumen contents were evacuated and weighed, papillae were collected from 3 locations in the rumen, and subsequently a VFA absorption test was performed. In the PG, mean rumen pH and molar acetate proportions decreased, and molar butyrate proportions increased continuously over the course of the trial, which can most likely be ascribed to an increased intake of rapidly fermentable carbohydrates. During the first weeks on a full grazing ration (wk 5-7), variation of rumen pH decreased, and in wk 5 a lower rumen content, papillae surface area, and potential for VFA absorption were observed. In wk 8 to 10, variation of rumen pH and total VFA concentrations increased again, and acetate/propionate ratio decreased. In wk-10 rumen content, papillae area and VFA absorption characteristics similar to initial levels were observed. Although continuous rumen pH assessments and LPS concentrations did not reveal an increased risk for subacute rumen acidosis (SARA) during the adaption period, histopathology of rumen papillae and potential for VFA absorption indicated a possible risk for rumen health. An increased risk for SARA was observed in wk 9 and 10 in the PG, but rumen LPS concentrations and histopathology were not adversely affected. Results of the present study suggest that after behavioral and metabolic adaptation to the transition from a TMR to a pasture-based ration, no adverse effects on rumen morphology and absorption capacity occurred, although rumen pH after adaptation to pasture indicated increased risk of SARA.

Glucose transporter expression differs between bovine monocyte and macrophage subsets and is influenced by milk production.

The peripartal period of dairy cows is characterized by negative energy balance and higher incidences of infectious diseases such as mastitis or metritis. With the onset of lactation, milk production is prioritized and large amounts of glucose are transported into the mammary gland. Decreased overall energy availability might impair the function of monocytes acting as key innate immune cells, which give rise to macrophages and dendritic cells and link innate and adaptive immunity. Information on glucose requirements of bovine immune cells is rare. Therefore, this study aims to evaluate glucose transporter expression of the 3 bovine monocyte subsets (classical, intermediate, and nonclassical monocytes) and monocyte-derived macrophages and to identify influences of the peripartal period. Blood samples were either collected from nonpregnant healthy cows or from 16 peripartal German Holstein cows at d -14, +7, and +21 relative to parturition. Quantitative real-time PCR was applied to determine mRNA expression of glucose transporters (GLUT) 1, GLUT3, and GLUT4 in monocyte subsets and monocyte-derived macrophages. The low GLUT1 and GLUT3 expression in nonclassical monocytes was unaltered during differentiation into macrophages, whereas in classical and intermediate monocytes GLUT expression was downregulated. Alternatively activated M2 macrophages consumed more glucose compared with classically activated M1 macrophages. The GLUT4 mRNA was only detectable in unstimulated macrophages. Neither monocytes nor macrophages were insulin responsive. In the peripartum period, monocyte GLUT1 and GLUT3 expression and the GLUT3/GLUT1 ratio were negatively correlated with lactose production. The high-affinity GLUT3 transporter appears to be the predominant glucose transporter on bovine monocytes and macrophages, especially in the peripartal period when blood glucose levels decline. Glucose transporter expression in monocytes is downregulated as a function of lactose production, which might impair monocyte to macrophage differentiation.

Metabolic and clinical response to Escherichia coli lipopolysaccharide in layer pullets of different genetic backgrounds supplied with graded dietary L-arginine.

L-arginine (Arg) is an essential amino acid in birds that plays a decisive role in avian protein synthesis and immune response. Effects of graded dietary Arg supply on metabolic and clinical response to Escherichia coli lipopolysaccharide (LPS) were studied over 48 hours after a single intramuscular LPS injection in 18-week-old genetically diverse purebred pullets. LPS induced a genotype-specific fever response within 4 hours post injectionem. Whereas brown genotypes showed an initial hypothermia followed by longer-lasting moderate hyperthermia, white genotypes exhibited a biphasic hyperthermia without initial hypothermia. Furthermore, within 2 hours after LPS injection, sickness behavior characterized by lethargy, anorexia, intensified respiration, and ruffled feathers appeared, persisted for 3 to 5 hours and recovered 12 hours post injectionem. The varying grades of Arg did not alter the examined traits named above, whereas insufficient Arg reduced body growth and increased relative weights of liver and pancreas significantly. At 48 hours post injectionem, increased relative weights of liver and spleen were also found in LPS treated pullets, whereas LPS decreased those of pancreas, bursa, thymus, and cecal tonsils. Moreover, LPS lowered the sum of plasma amino acids and decreased plasma concentrations of Arg, citrulline, glutamate, methionine, ornithine, phenylalanine, proline, tryptophan, and tyrosine, and increased those of aspartate, glutamine, lysine, 1- and 3-methyl-histidine. Elevating concentrations of dietary Arg led to increasing plasma concentrations of Arg, citrulline, ornithine, and 3-methyl-histidine subsequently. As quantitative expression of LPS-induced anorexia, proteolysis, and the following changes in plasma amino acids, pullets showed a significant decrease of feed and nitrogen intake and catabolic metabolism characterized by negative nitrogen balance and body weight loss in the first 24 hours post injectionem. Pullets recovered from the challenge within the second 24 hours post injectionem and changed to anabolism with re-increased feed and nitrogen intake, positive nitrogen retention, and weight gain. To conclude, present results confirmed that LPS induced numerous metabolic and physiological changes in pullet's genotypes, whereas dietary Arg affected the examined traits only slightly.

Effect of monensin and essential oils on performance and energy metabolism of transition dairy cows.

This work examined preventive effects of a dietary and a medical intervention measure on postpartum (p.p.) ketogenesis in dairy cows overconditioned in late pregnancy. Sixty German Holstein cows were allocated 6 weeks antepartum (a.p.) to three high body condition score (BCS) groups (BCS 3.95 ± 0.08) and one low BCS group (LC, BCS 2.77 ± 0.14). Concentrate proportion in diet a.p. was higher (60% vs. 20%) and increase in proportion p.p. from 30% up to 50% decelerated (3 vs. 2 weeks) in high BCS groups. High BCS cows received a monensin controlled-release capsule (CRC) (HC/MO), a blend of essential oils (HC/EO) or formed a control group (HC). Performance parameters and energy status were evaluated in three periods [day (d) -42 until calving, one until 14 days in milk (DIM), 15 until 56 DIM]. Feed efficiency was 65% and 53% higher in HC/MO than in LC (p < 0.001) and HC groups (p = 0.002) in the second period. Milk fat content was higher in HC/EO (5.60 vs. 4.82%; p = 0.012) and milk urea higher in HC/MO (135 mg/kg) than in LC cows (107 mg/kg; p < 0.001). Increased p.p. levels of non-esterified fatty acids in serum were found in HC (p = 0.003), HC/MO (p = 0.068) and HC/EO (p = 0.002) in comparison with LC cows. Prevalence of subclinical and clinical ketosis was 54% and 46%, respectively, in HC group. Monensin decreased the prevalence to 50% and 7% respectively. Ruminal fermentation pattern showed higher proportions of propionate (23.43 mol % and 17.75 mol %, respectively; p < 0.008) and lower acetate:propionate ratio (2.66 vs. 3.76; p < 0.001) in HC/MO than HC group. Results suggest that a monensin CRC improved energy status and feed efficiency of transition dairy cows while essential oils failed to elicit any effect.