Short-term exposure of dairy cows to pyrrolizidine alkaloids from tansy ragwort (Jacobaea vulgaris Gaertn.): effects on organs and indicators of energy metabolism.

Preserved feed from meadows contaminated with ragwort (Jacobaea vulgaris, Gaertn.) may expose livestock to pyrrolizidine alkaloids (PA). Dairy cows are considered to be very susceptible animals and a PA ingestion can lead to liver and further organ damages and even death. Due to the lack of data, the present study aimed to evaluate critical PA doses based on organ effects, with a special focus on liver lesions and on indicators of energy metabolism. Therefore, 16 dairy cows (n = 4 per group) were exposed to increasing PA doses (group: CONMolasses: <0.001 mg PA/kg body weight (BW)/day (d); PA1: 0.47 mg PA/kg BW/d; PA2: 0.95 mg PA/kg BW/d; PA3: 1.91 mg PA/kg BW/d) for 28 days. Constant dosing was ensured by a defined PA extract administered orally once daily. Histological examinations of the livers showed infiltration by immune cells, higher proportions of apoptotic cells and enlargement of hepatocyte nuclei in the highest exposed group. In addition, bile volume increased with PA dose, which may indicate a cholestasis. Despite the signs of incipient liver damage, liver lipid content and clinical chemical parameters related to energy metabolism, such as glucose, non-esterified fatty acids and ßhydroxybutyrate, remained unaffected. Fat depot masses were also not significantly altered over time, suggesting that PA exposure did not induce a wasting syndrome. The liver showed slight microscopic changes already at a dosage of 0.95 mg PA/kg BW/d. However, the short-term metabolic indicators of energy status, lipolysis and ketogenesis, glucose, NEFA and BHB, as well as changes in fat depot, which serves as a longer-term indicator of lipolysis, remained unaffected in all treatment groups in the chosen scenario. These findings suggest that despite histopathological and clinical-chemical evidence of PA-associated hepatocellular lesions, liver function was not compromised.

Influences of dietary-reduced nitrogen (N) and phosphorus (P) on chemical body composition, N and P retention and health traits of contemporary barrows.

Farmgate balances are used as a tool for monitoring nutrient surpluses at farm level. In Germany, preparation of farmgate balances is legally mandatory and also requires data on chemical body composition, especially concentration of nitrogen (N) and phosphorus (P), of farm animals. It is well known that increased N and P efficiency results in lowered N and P excretions with the manure and therefore mitigates negative consequences of high N and P release into the environment (e.g. eutrophication of surface waters), especially in areas with high livestock density. In this context, feeding N- and P-reduced diets can be a strategy for increasing N and P efficiency in fattening pigs. To investigate the influence of N- and P-reduced diets on chemical body composition of barrows and to update current used data basis, 8 barrows were slaughtered after a balance trial and their bodies were subjected to full body analysis. During the balance trial, pigs received the control diet (CON) meeting common nutrient requirements or the N- and P-reduced diet (NPred) in a three-phased feeding regimen (n = 4/diet, 3 weeks/phase). Pigs were slaughtered with an average live weight (LW) of 123.3 ± 7.5 kg and carcasses were manually dissected in four fractions. Fractions were analysed for nutrient concentration. Furthermore, organs were weighed individually and blood serum was sampled during exsanguination. Serum samples were analysed for clinical-chemical traits. Chemical body composition did not significantly differ between NPred- and CON-fed pigs. N concentration was 23.3 ± 0.3 and 24.5 ± 1.0 g/kg, P concentration was 5.2 ± 0.1 and 5.5 ± 0.4 g/kg in the empty body of NPred- and CON-fed pigs (p = 0.073, 0.164). N and P retention between the experiment's start and slaughter did not differ between the feeding groups (p = 0.641, 0.240). Variables related to liver integrity, energy metabolism and electrolytes were similar between CON- and NPred-fed pigs. Traits related to protein metabolism showed significantly reduced concentrations of urea and albumin in NPred-fed pigs (p = 0.013, 0.025), but no hypoalbuminaemia. Results suggest that N- and P-reduced feeding does not significantly affect chemical body composition of contemporary barrows.

Effects of dietary-reduced nitrogen (N) and phosphorus (P) on N and P balance, retention and nutrient digestibility of contemporary fattening pigs fed ad libitum.

The reduction of nitrogen (N) and phosphorus (P) in fattening pigs' diets is one possible approach to lower N and P excretion in livestock farming relative to N and P intake. Due to the implementation of the European Nitrates Directive and the consecutive amendments to the German fertiliser legislation since 2017, N- and P-reduced diets for fattening pigs are becoming more and more important and are increasingly used in practice. To investigate the effects of such diets on N and P balance and retention as well as on nutrient digestibility of contemporary fattening pigs, a balance experiment was performed with eight barrows (average live weight = 61.5 ± 2.1 kg) which were surgically fitted with a simple T-cannula at the terminal ileum. The pigs received a control diet meeting nutrient requirements (CON) and an N- and P-reduced diet (NPred) ad libitum (n = 4/diet) in a 3-phased feeding regimen (3 weeks/phase). In the last week of each phase, faeces and urine were collected quantitatively for 5 days followed by a 2 × 12 hours collection of ileal digesta. Daily feed intake, live weight gain and feed-to-gain ratio did not differ between CON and NPred. NPred-fed pigs consumed 10.5% (p = 0.006) and excreted 28.3% (p = 0.028) less N than CON-fed pigs. Phosphorus excretion was lowered by 15.1% in NPred-fed pigs (p = 0.012). N and P retention did not differ between CON and NPred, but were elevated in comparison to other studies. N and P efficiency, expressed as nutrient retention divided by nutrient intake, was higher in NPred - than CON-fed pigs (N: 68 vs 60%, P: 54.2 vs 49.3%). Apparent post-ileal digestibility coefficient (DCpost-ileal) and apparent total tract digestibility coefficient (DCtotal) of crude protein were higher in NPred - than CON-fed pigs (p < 0.013), but apparent precaecal digestibility coefficient (DCpc) of crude protein was unaffected by diet. DCpc, DCpost-ileal and DCtotal of P were similar for CON- and NPred-fed pigs. NPred-fed pigs showed an elevated DCpc and DCtotal of organic matter, N-free-extractives and starch compared to CON-fed pigs. DCpc of calcium was also higher in NPred-fed pigs. In conclusion, the results suggest that N- and P-reduced feeding of fattening pigs remains an effective strategy to lower the N and P release into the environment. Furthermore, results indicate that N- and P-reduced feeding leads to a higher N and P efficiency in contemporary fattening pigs.

Short-term exposure of dairy cows to pyrrolizidine alkaloids from tansy ragwort (Jacobaea vulgaris Gaertn.): effects on health and performance.

The increasing spread of ragworts is observed with concern. Ragworts like tansy ragwort (Jacobaea vulgaris Gaertn.) or marsh ragwort (J. aquatica) contain pyrrolizidine alkaloids (PA) which may induce hepatotoxic effects. Grazing animals usually avoid ragworts if their pasture management is appropriate. Preserved feed prepared from ragworts contaminated meadows may, however, lead to a significant exposure to PA. Previous studies on toxicity of PA for dairy cows revealed inconsistent results due to feeding ragwort plant material which was associated with heterogeneous PA exposure and thus failed to conclusively deduce critical PA doses. Therefore, the aim of the present study was to expose dairy cows (n = 4 per group) in a short-term scenario for 28 days with increasing PA doses (PA1: 0.47 mg PA/kg body weight (BW)/day (d); PA2: 0.95 mg PA/kg BW/d; PA3: 1.91 mg PA/kg BW/d) via oral administration by gavage of a defined PA-extract. While group PA3 was dosed with the PA-extract alone, groups PA2 and PA1 received PA-extracts blended in similar volumes with molasses to provide comparable amounts of sugar. Additionally, two control groups were treated either with water (CONWater) or with molasses (CONMolasses) to assess the effects of sugar without PA interference. While clinical traits including dry matter intake, milking performance, rectal body temperature, ruminal activity and body condition score (BCS) were not influenced by PA exposure, activities of enzymes indicative for liver damages, such as gamma-glutamyltransferase (GGT), aspartate aminotransferase (AST) and glutamate dehydrogenase (GLDH), increased significantly over time at an exposure of 1.91 mg total PA/kg BW/d.

Estimates of Water Consumption of Growing Pigs under Practical Conditions Based on Climate and Performance Data.

The water consumption of fattening pigs was recorded under practical conditions and compared with calculated water consumption. The experiment was carried out in the summer of 2020 with 79 fattening pigs. Data loggers were used to record the climate data, such as temperature and relative humidity. These data were used to calculate the temperature-humidity index (THI). It was found that there were sometimes considerable discrepancies between the measured and the calculated water consumption. One possible reason for this discrepancy could be the age of the existing water requirement equations, as in recent decades there has been a clear breeding development and thus a strong increase in pig performance. Based on these deviations, six new water consumption equations were established, which considered the variables body weight (BW), temperature, THI and feed consumption. It was found that the THI and BW should be included in one equation as predictor variables and the evaluation also showed good results. Its use, in practice, should also be considered. Overall, it became apparent that there is still a need for further research to make water consumption equations more precise. This would require a larger database.

Inactivation of zearalenone (ZEN) and deoxynivalenol (DON) in complete feed for weaned piglets: Efficacy of ZEN hydrolase ZenA and of sodium metabisulfite (SBS) as feed additives.

Female pigs respond sensitive both to DON and ZEN with anorexia and endocrine disruption, respectively, when critical diet concentrations are exceeded. Therefore, the frequent co-contamination of feed by DON and ZEN requires their parallel inactivation. The additive ZenA hydrolyzes ZEN while SBS inactivates DON through sulfonation. Both supplements were simultaneously added (+, 2.5 g SBS and 100 U ZenA/kg) to a control diet (CON-, 0.04 mg DON and < 0.004 mg ZEN/kg; CON+, 0.03 mg DON and < 0.004 mg ZEN/kg) and a Fusarium toxin contaminated diet (FUS-, 2.57 mg DON and 0.24 mg ZEN/kg; FUS+, 2.04 mg DON and 0.24 mg ZEN/kg). The 4 diets were fed to 20 female weaned piglets each (6 kg initial body weight) for 35 days; the piglets were sacrificed thereafter for collecting samples. Supplements improved performance and modified metabolism and hematology independent of dietary DON contamination. The mechanisms behind these changes could not be clarified and require further consideration. SBS reduced DON concentration in feed by approximately 20% and to the same extent in blood plasma and urine suggesting that no further DON sulfonate formation occurred in the digestive tract before absorbing DON in the upper digestive tract or that additionally formed DON sulfonates escaped absorption. DON sulfonates were detected in feces suggesting that unabsorbed DON sulfonates reached feces and/or that unabsorbed DON was sulfonated in the hindgut. The observed reduction rate of 20% was evaluated to be insufficient for feeding practice. Galenic form of SBS added to dry feed needs to be improved to support the DON sulfonation in the proximal digestive tract.ZenA was active in the digestive tract as demonstrated by the presence of its hydrolyzed none-estrogenic reaction products hydrolyzed ZEN (HZEN) and decarboxylated and hydrolyzed ZEN (DHZEN) both in feces, systemic circulation, and urine of group FUS+ compared to group FUS-. The presence of these hydrolysis products was paralleled by a significant decrease in high-estrogenic ZEN concentrations which, in turn, was related to a decrease in relative weights of uteri and ovaries when compared to group FUS-. Thus, ZenA was proven to be effective; both in terms of biomarkers and biological effects.

Deoxynivalenol affects cell metabolism in vivo and inhibits protein synthesis in IPEC-1 cells.

Deoxynivalenol is present in forage crops in concentrations that endanger animal welfare but is also found in cereal-based food. The amphipathic nature of mycotoxins allows them to cross the cell membrane and interacts with different cell organelles such as mitochondria and ribosomes. In our study, we investigated the gene expression of several genes in vivo and in vitro that are related to the metabolism. We observed a significantly higher COX5B and MHCII expression in enterocytes of DON-fed pigs compared to CON-fed pigs and a marked increase in GAPDH and SLC7A11 in DON-fed pigs, but we could not confirm this in vitro in IPEC-1. In vitro, functional metabolic analyses were performed with a seahorse analyzer. A significant increase of non-mitochondrial respiration was observed in all DON-treatment groups (50-2000 ng/mL). The oxygen consumption of cells, which were cultured on membranes, was examined with a fiber-glass electrode. Here, we found significantly lower values for DON 200- and DON 2000-treatment group. The effect on ribosomes was investigated using biorthogonal non-canonical amino acid tagging (BONCAT) to tag newly synthesized proteins. A significantly reduced amount was found in almost all DON-treatment groups. Our findings clearly show that apical and basolateral DON-treatment of epithelial cell layer results in decreasing amounts of newly synthesized proteins. Furthermore, our study shows that DON affects enterocyte metabolism in vivo and in vitro.

Does the Implementation of an Animal Welfare Programme on a Farm Yield a Demonstrable Improvement in Fattening Pig Welfare?

In the course of social criticism of fattening pig farming, an animal welfare programme called "Initiative Animal Welfare" (ITW) was founded to increase animal welfare in pig farming in Germany. Furthermore, there is a legal obligation to record animal welfare parameters as a self-monitoring measure. The "German Association for Technology and Structures in Agriculture" published a guideline on the applicable animal welfare criteria. This guide formed the basis of this study's data collection. The aim was to apply the animal welfare parameters on farms by comparing the results between farms participating in ITW with those not participating. A cumulative score was calculated by evaluating the collected data. In addition, the relative risk was calculated in order to estimate the risk of finding a negative expression of a parameter. Our data show that ITW farms did not perform significantly better than the farms without ITW in terms of both the cumulative score and the relative risk. Overall, it must be considered that in both farm variants the occurrence of negative evaluations was very rare and the visited farms thus certainly can be considered to be well-managed farms. Climate parameters were recorded in each compartment and showed no significant differences in most cases.

Dietary L-Carnitine Affects Leukocyte Count and Function in Dairy Cows Around Parturition.

In early lactation, an energy deficit leading to a negative energy balance (NEB) is associated with increased susceptibility to disease and has been shown to be an important factor during transition in dairy cows. L-carnitine as a key factor in the mitochondrial transport of fatty acids and subsequently for ß-oxidation and energy release is known to modulate mitochondrial biogenesis and thus influence metabolism and immune system. In the current study, we characterized hematological changes around parturition and investigated the potential effects of dietary L-carnitine supplementation on immune cell functions. For this approach, dairy cows were assigned either to a control (CON, n = 30) or an L-carnitine group [CAR, n = 29, 25 g rumen-protected L-carnitine per cow and day (d)]. Blood samples were taken from d 42 ante partum (ap) until d 110 post-partum (pp), with special focus and frequent sampling from 0.5 to72 h post-calving to clarify the impact of L-carnitine supplementation on leukocyte count, formation of reactive oxygen species (ROS) in polymorphonuclear cells (PMN) and peripheral mononuclear cells (PBMC) and their phagocytosis activity. Blood cortisol concentration and the capacity of PBMC proliferation was also investigated. All populations of leukocytes were changed during the peripartal period, especially granulocytes showed a characteristic increase up to 4 h pp. L-carnitine supplementation resulted in increased levels of eosinophils which was particularly pronounced one day before to 4 h pp, indicating a possible enhanced support for tissue repair and recovery. Non-supplemented cows showed a higher phagocytic activity in PBMC as well as a higher phagocytic capacity of PMN during the most demanding period around parturition, which may relate to a decrease in plasma levels of non-esterified fatty acids reported previously. L-carnitine, on the other hand, led to an increased efficiency to form ROS in stimulated PMN. Finally, a short period around calving proved to be a sensitive period in which L-carnitine administration was effective.

Correction: Schwarzkopf et al. Weaning Holstein Calves at 17 Weeks of Age Enables Smooth Transition from Liquid to Solid Feed. Animals 2019, 9, 1132.

The authors wish to make the following correction to their paper [...].

Does chronic dietary exposure to the mycotoxin deoxynivalenol affect the porcine hepatic transcriptome when an acute-phase response is initiated through first or second-pass LPS challenge of the liver?

The sensitivity of pigs to deoxynivalenol (DON) might be increased by systemic inflammation (SI), which also has consequences for hepatic integrity. Liver lesions and a dys-regulated gene network might hamper hepatic handling and elimination of DON whereby the way of initiation of hepatic inflammation might play an additional role. First and second-pass exposure of the liver with LPS for triggering a SI was achieved by LPS infusion via pre- or post-hepatic venous route, respectively. Each infusion group was pre-conditioned either with a control diet (0.12 mg DON/kg diet) or with a DON-contaminated diet (4.59 mg DON/kg diet) for 4 wk. Liver transcriptome was evaluated at 195 min after starting infusions. DON exposure alone failed to modulate the mRNA expression significantly. However, pre- and post-hepatic LPS challenges prompted transcriptional responses in immune and metabolic levels. The mRNAs for B-cell lymphoma 2-like protein 11 as a key factor in apoptosis and IFN-γ released by T cells were clearly up-regulated in DON-fed group infused with LPS post-hepatically. On the other hand, mRNAs for nucleotide binding oligomerization domain containing 2, IFN-α and eukaryotic translation initiation factor 2α kinase 3 as ribosomal stress sensors were exclusively up-regulated in control pigs with pre-hepatic LPS infusion. These diverse effects were traced back to differences in TLR4 signalling.

Dietary l-carnitine Supplementation Modifies the Lipopolysaccharide-Induced Acute Phase Reaction in Dairy Cows.

l-carnitine plays an important role in energy metabolism through supporting the transport of activated fatty acids to the subcellular site of ß-oxidation. An acute phase reaction (APR) is known as an energy consuming process. Lipopolysaccharides (LPS) are often used in animal models to study intervention measures during innate immune responses such as APR. Thus, the aim of the study was to investigate the effects of dietary l-carnitine supplementation during an LPS-induced APR in mid-lactating German Holstein cows. Animals were assigned to a control (CON, n = 26) or l-carnitine group (CAR, n = 27, 25 g rumen-protected l-carnitine/cow/d) and received an intravenous injection of LPS (0.5 µg/kg body weight) at day 111 post-partum. Blood samples were collected from day 1 pre-injection until day 14 post-injection (pi). From 0.5 h pi until 72 h pi blood samplings and clinical examinations were performed in short intervals. Clinical signs of the APR were not altered in group CAR except rumen motility which increased at a lower level compared to the CON group after a period of atonia. Group CAR maintained a higher insulin level compared to group CON even up to 72 h pi which might support glucose utilization following an APR.

Sodium sulfite (SoS) as decontamination strategy for Fusarium-toxin contaminated maize and its impact on immunological traits in pigs challenged with lipopolysaccharide (LPS).

The main objective of this study was to evaluate the effects of sodium sulfite (SoS) treatment of maize and its impact on the porcine immune system in the presence of an LPS-induced systemic inflammation. Control maize (CON) and Fusarium-toxin contaminated maize (FUS) were wet-preserved (20% moisture) for 79 days with (+) or without (-) SoS and then included at 10% in a diet, resulting in four experimental groups: CON-, CON+, FUS-, and FUS+ with deoxynivalenol (DON) concentrations of 0.09, 0.05, 5.36, and 0.83 mg DON/kg feed, respectively. After 42-day feeding trial (weaned barrows, n = 20/group), ten pigs per group were challenged intraperitoneally with either 7.5 µg LPS/kg BW or placebo (0.9% NaCl), observed for 2 h, and then sacrificed. Blood, mesenteric lymph nodes, and spleen were collected for phenotyping of different T cell subsets, B cells, and monocytes. Phagocytic activity and intracellular formation of reactive oxygen species (ROS) were analyzed in both polymorphonuclear cells (PMN) and peripheral blood mononuclear cells (PBMC) using flow cytometry. Our results revealed that the impact of DON was more notable on CD3+CD4+CD8+ T cells in lymphoid tissues rather than in blood T cells. In contrast, SoS treatment of maize altered leukocyte subpopulations in blood, e.g., reduced the percentage and fluorescence signal of CD8high T cells. Interestingly, SoS treatment reduced the amount of free radicals in basal ROS-producing PMNs only in LPS-challenged animals, suggesting a decrease in basal cellular ROS production (pSoS*LPS = 0.022).

Oral exposure of pigs to the mycotoxin deoxynivalenol does not modulate the hepatic albumin synthesis during a LPS-induced acute-phase reaction.

The sensitivity of pigs to deoxynivalenol (DON) might be influenced by systemic inflammation (SI) which impacts liver. Besides following acute-phase proteins, our aim was to investigate both the hepatic fractional albumin (ALB) synthesis rate (FSR) and the ALB concentration as indicators of ALB metabolism in presence and absence of SI induced by LPS via pre- or post-hepatic venous route. Each infusion group was pre-conditioned either with a control diet (CON, 0.12 mg DON/kg diet) or with a DON-contaminated diet (DON, 4.59 mg DON/kg diet) for 4 wk. A depression of ALB FSR was observed 195 min after LPS challenge, independent of feeding group or LPS application route, which was not paralleled by a down-regulated ALB mRNA expression but by a reduced availability of free cysteine. The drop in ALB FSR only partly explained the plasma ALB concentrations which were more depressed in the DON-pre-exposed groups, suggesting that ALB levels are influenced by further mechanisms. The abundances of haptoglobin, C-reactive protein, serum amyloid A, pig major acute-phase protein, fibrinogen and LPS-binding protein mRNA were up-regulated upon LPS stimulation but not accompanied by increases in the plasma concentrations of these proteins, pointing at an imbalance between synthesis and consumption.

Effects of a Dietary L-Carnitine Supplementation on Performance, Energy Metabolism and Recovery from Calving in Dairy Cows.

Dairy cows are metabolically challenged during the transition period. Furthermore, the process of parturition represents an energy-consuming process. The degree of negative energy balance and recovery from calving also depends on the efficiency of mitochondrial energy generation. At this point, L-carnitine plays an important role for the transfer of fatty acids to the site of their mitochondrial utilisation. A control (n = 30) and an L-carnitine group (n = 29, 25 g rumen-protected L-carnitine per cow and day) were created and blood samples were taken from day 42 ante partum (ap) until day 110 post-partum (pp) to clarify the impact of L-carnitine supplementation on dairy cows, especially during the transition period and early puerperium. Blood and clinical parameters were recorded in high resolution from 0.5 h to 72 h pp. L-carnitine-supplemented cows had higher amounts of milk fat in early lactation and higher triacylglyceride concentrations in plasma ap, indicating increased efficiency of fat oxidation. However, neither recovery from calving nor energy balance and lipomobilisation were influenced by L-carnitine.

Weaning Holstein Calves at 17 Weeks of Age Enables Smooth Transition from Liquid to Solid Feed.

Development of calves depends on prenatal and postnatal conditions. Primiparous cows were still maturing during pregnancy, which can lead to negative intrauterine conditions and affect the calf's metabolism. It is hypothesized that weaning calves at higher maturity has positive effects due to reduced metabolic stress. We aimed to evaluate effects of mothers' parity and calves' weaning age on growth performance and blood metabolites. Fifty-nine female Holstein calves (38.8 ± 5.3 kg birth weight, about 8 days old) were used in a 2 × 2 factorial experiment with factors weaning age (7 vs. 17 weeks) and parity of mother (primiparous vs. multiparous cows). Calves were randomly assigned one of these four groups. Live weight, live weight gain and morphometry increased over time and were greater in calves weaned later. Metabolic indicators except total protein were interactively affected by time and weaning age. Leptin remained low in early-weaned calves born to primiparous cows, while it increased in the other groups. The results suggest that weaning more mature calves has a positive effect on body growth, and calves born to primiparous cows particularly benefit from this weaning regimen. It also enables a smooth transition from liquid to solid feed, which might reduce the associated stress of weaning.

Kinetic studies on clinical and immunological modulations by intramuscular injection of Escherichia coli LPS in laying hens.

The present study investigated clinical and immunological modulations due to intramuscular injection of Escherichia coli LPS in 49-wk-old laying hens over 48 h post injection (p.i.). LPS induced characteristic sickness behavior but no significant body temperature alterations ( P > 0.05). During experimental period decreases in blood albumin, calcium, phosphorus and tryptophan concentrations, hyperglycemia, increased plasma nitrite concentrations, leucopenia, decreased thrombocyte counts, lymphopenia, heterophilia and an increased heterophilic granulocyte/lymphocyte (H/L) ratio were observed after LPS administration. Time-dependent effects were shown on T and B cell subsets in caecal tonsils (CT) and on splenic CD3+/CD4+/CD8+ proportions, on IL-1ß and -10 and inducible NO synthase mRNA expression in peripheral blood lymphocytes (PBL), liver, spleen and CT, and on the mRNA expression of the TLR4 in PBL, liver and spleen p.i. ( P < 0.05). The main responding period of mentioned alterations due to LPS appears to include the period from 2 until 8 h p.i. According to the H/L ratio, the most stressful phase was 5 h p.i. T and B cell subsets in CT, the IL-1ß and TLR4 mRNA expression in liver and plasma nitrite concentrations seemed to be affected for a longer period.

Analytical method for the determination of polyethylenglycole 400 as marker in porcine plasma.

Polyethylenglycole (PEG) is a widespread linear polymer which can be utilized as a solute digestive and intestinal permeability marker in nutritional physiology studies depending on chain length/molecular mass. PEG 400 is proposed to be an ideal permeability marker. Due to its molecular mass (238-590 g/mol) and characteristics, PEG 400 is suggested to be used as a surrogate for studying the paracellular permeability of small hydrophilic molecules. For this purpose, a liquid chromatographic-tandem mass spectrometric method has been developed for the determination of the major oligomers of PEG 400 in porcine plasma. The analysis included a simple and rapid clean-up step where proteins were precipitated. The most intense ions corresponding to seven PEG 400 oligomers were separated within 7 min. Validation of the optimized method was performed in the range of 500-18,000 ng/mL. Mean recoveries between 93 and 105% were achieved using spiked plasma samples in three different concentration levels. The limit of quantification ranged between 11 and 244 ng/mL. The applicability of the method was demonstrated by the analysis of porcine plasma samples obtained from an animal experiment with barrows. The kinetic course of administrated PEG 400 was shown based on the dataset of two barrows selected from the control group, and it was figured out that relative proportion of each PEG oligomer in portal plasma decreased with increasing molecular mass.

Effects of deoxynivalenol-feed contamination on circulating LPS in pigs.

Low concentration of LPS can be detected in healthy mammals without triggering systemic inflammation. Here we analysed the influence of the mycotoxin deoxynivalenol (DON) on very low LPS concentrations and the role of DON in the physiology of pigs challenged with high artificial LPS dosage mimicking septic shock. Pigs were fed for 29 d with DON-contaminated (4.59 mg/kg feed) or control feed. Samples of control animals showed 6.6 ± 13.5 pg/ml LPS in portal and 3.1 ± 7.6 pg/ml LPS in jugular serum samples. In the DON fed group, 3.4 ± 7.2 pg/ml and 0.6 ± 0.8 pg/ml were detected. The differences were statistically not significant, indicating that DON is not a trigger for enhanced LPS transfer into the blood circulation. Next, pigs were challenged with 7.5 µg LPS/kg body mass via portal or jugular route. The application route did not significantly influence the LPS concentration. We expected higher circulating LPS concentrations in the presence of DON due to the additional stress of liver metabolism and reduced liver capacity to remove LPS from circulation. This scenario is supported by tendency. In summary, we found that DON is unlikely to influence LPS transfer in the gut; DON likely reduces the capacity for LPS removal in septic shock conditions.

Effects of a Fusarium Toxin-Contaminated Maize Treated with Sodium Sulfite on Male Piglets in the Presence of an LPS-Induced Acute Inflammation.

We investigated the effects of feeding sodium sulfite (SoS) treated uncontaminated and Fusarium contaminated maize in a porcine lipopolysaccharide (LPS) challenge model. Eighty piglets (7.59 ± 0.92 kg body weight [BW]) were equally assigned to one of four experimental diets containing 10% maize, either uncontaminated and untreated (CON-, 0.09 mg deoxynivalenol [DON]/kg diet) or uncontaminated and SoS-treated (CON+, wet-preserved with 5 g SoS/kg maize; 0.05 mg DON/kg diet), or prepared with 10% of a Fusarium contaminated maize containing mainly deoxynivalenol (DON), either contaminated and untreated (FUS-, 5.36 mg DON/kg diet), or contaminated and SoS-treated (FUS+, wet-preserved with 5 g SoS/kg maize; 0.83 mg DON/kg diet). At day 42 of experiment, ten pigs of each group were injected intraperitoneally with either 7.5 µg LPS/kg BW or placebo (0.9% NaCl). At 120 min after injection, blood samples were collected to analyse TNF-α, hematological profile, clinical biochemistry as well as the redox status. A significant increase in body temperature and cytokine TNF-α concentration was observed in the LPS-injected piglets. Results for hematology, clinical chemistry and redox status indicate no effects of SoS treatment, with exception of neutrophil counts being significantly more pronounced after feeding the SoS treated FUS maize. In conclusion, SoS treatment of maize did not modulate the LPS-induced acute inflammation.