Beyond the microcirculation: sequestration of infected red blood cells and reduced flow in large draining veins in experimental cerebral malaria.

Sequestration of infected red blood cells (iRBCs) in the microcirculation is a hallmark of cerebral malaria (CM) in post-mortem human brains. It remains controversial how this might be linked to the different disease manifestations, in particular brain swelling leading to brain herniation and death. The main hypotheses focus on iRBC-triggered inflammation and mechanical obstruction of blood flow. Here, we test these hypotheses using murine models of experimental CM (ECM), SPECT-imaging of radiolabeled iRBCs and cerebral perfusion, MR-angiography, q-PCR, and immunohistochemistry. We show that iRBC accumulation and reduced flow precede inflammation. Unexpectedly, we find that iRBCs accumulate not only in the microcirculation but also in large draining veins and sinuses, particularly at the rostral confluence. We identify two parallel venous streams from the superior sagittal sinus that open into the rostral rhinal veins and are partially connected to infected skull bone marrow. The flow in these vessels is reduced early, and the spatial patterns of pathology correspond to venous drainage territories. Our data suggest that venous efflux reductions downstream of the microcirculation are causally linked to ECM pathology, and that the different spatiotemporal patterns of edema development in mice and humans could be related to anatomical differences in venous anatomy.

Deoxynivalenol triggers the expression of IL-8-related signaling cascades and decreases protein biosynthesis in primary monocyte-derived cells.

Humans and their immune system are confronted with mold-contaminated food and/or mold-contaminated air in daily life and indoor activities. This results in metabolic stress and unspecific disease symptoms. Other studies provided evidence that exposure to mold is associated with the etiology of allergies. Deoxynivalenol (DON) is of great concern due to its frequent occurrence in toxically relevant concentrations. The exposure to this toxin is a permanent health risk for both humans and farm animals because DON cannot be significantly removed during standard milling and processing procedures. However, the direct effect on immunity or hematology is poorly defined because most investigations could not separate the effect of DON-contaminated feed intake. Due to the widespread distribution of DON after rapid absorption, it is not surprising that DON is known to affect the immune system. The immune system of the organism has one important function, to defend against the invasion of unknown substances/organisms. This study shows for the first time a synergistic effect of both-low physiological DON-doses in combination with low LPS-doses with the focus on the IL-8 expression on protein and RNA level. Both doses were found in vivo. IL-8 together with other anorectic cytokines like IL-1ß can affect the food intake and anorexia. We could also show that a calcium-response is not involved in the increased IL-8 production after acute DON stimulation with high or low concentrations.

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.

Dietary fiber and its role in performance, welfare, and health of pigs.

Dietary fiber (DF) is receiving increasing attention, and its importance in pig nutrition is now acknowledged. Although DF for pigs was frowned upon for a long time because of reductions in energy intake and digestibility of other nutrients, it has become clear that feeding DF to pigs can affect their well-being and health. This review aims to summarize the state of knowledge of studies on DF in pigs, with an emphasis on the underlying mode of action, by considering research using DF in sows as well as suckling and weaned piglets, and fattening pigs. These studies indicate that DF can benefit the digestive tracts and the health of pigs, if certain conditions or restrictions are considered, such as concentration in the feed and fermentability. Besides the chemical composition and the impact on energy and nutrient digestibility, it is also necessary to evaluate the possible physical and physiologic effects on intestinal function and intestinal microbiota, to better understand the relation of DF to animal health and welfare. Future research should be designed to provide a better mechanistic understanding of the physiologic effects of DF in pigs.

[Myofascial chronic pelvic pain in women : A retrospective evaluation of the effects of specific diagnostics and therapy]. / Myofaszial bedingte chronische Unterbauchschmerzen bei Frauen : Eine retrospektive Auswertung der Auswirkungen einer spezifischen Diagnostik und Therapie.

BACKGROUND: The prevalence of chronic pelvic pain of 11.8% in the general population underlines the importance of this disease. However, the specific diagnostics and therapy of the muscles of this region are not yet part of the standard examination. The following study examines the effects of specific diagnostics and therapy on myofascial chronic pelvic pain. OBJECTIVES: The effectiveness of targeted diagnostics and multimodal therapy in the context of chronic pelvic pain and the need for a complementary drug adjustment. MATERIALS AND METHODS: This study retrospectively evaluated the data of 59 patients, who were referred to a pain center for treatment-resistant chronic pelvic pain in the period from January 2012 to April 2017. The pain needed to be clearly identified as myofascial. A previous minimum duration of pain as well as previous operations or other treatment procedures did not constitute exclusion criteria. Previous traumatization was a reason for exclusion. Therapy components included manual therapeutic treatment, training in self-stretching exercises, medication with the active ingredients flupirtine or methocarbamol, as well as relaxation procedures. Therapy was evaluated on the basis of the German Pain Questionnaire. RESULTS: After the treatment interval, the following statistically significant improvements were recorded: The average pain intensity decreased by 29.95 points (standard deviation [SD] = 20.61). General quality of life (Marburg questionnaire on habitual well-being, MFHW) increased by 1.1 points (SD = 0.73). The depression and anxiety assessment decreased by 2.56 (SD = 3.99) and 2.63 points (SD = 5.21) respectively. CONCLUSION: A multimodal therapy concept with a manual therapeutic treatment focus can lead to an improvement in pain symptoms and quality of life in patients with myofascial chronic pelvic pain after a treatment period of 120 days. Myofascial syndromes of urogenital muscles must be considered in the assessment of the cause of chronic pelvic pain.

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.

Deoxynivalenol, but not E. coli lipopolysaccharide, changes the response pattern of intestinal porcine epithelial cells (IPEC-J2) according to its route of application.

The porcine intestinal epithelium is a primary target for mycotoxin deoxynivalenol (DON) and lipopolysaccharides (LPS). Although epithelial cells are exposed to these toxins mainly from the luminal-chyme compartment an exposure from the blood side resulting from systemic absorption cannot be excluded. Thus, we investigated the effect of DON and LPS, alone or combined, on porcine intestinal epithelial cells IPEC-J2 on a transcriptional, translational and functional level when administered either from apical or basolateral. IPEC-J2 cells were cultured on 12-well inserts in complete medium at 5% CO2 and 39°C and subjected to following treatments: control (CON), 2000 ng/mL DON, 1 µg/mL LPS or DON+LPS for 72 h, either from apical or basolateral. Transepithelial electrical resistance (TEER), protein and IL-8 content were measured and microarray analysis, qRT-PCR (IL-8, zonula occludens-1 ZO-1, ß-actin), Western Blot (ZO-1, ß-actin) and immunofluorescence (ZO-1) were performed. Data of at least three independent experiments were analysed with ANOVA and Dunnett's post hoc test. Basolateral DON resulted in significantly lower cell counts (p<0.05) with larger cells (p<0.01), whereas apical DON reduced total (p<0.001) and specific protein content (IL-8 content CON vs. DON: 2378 pg/3 mL vs. 991 pg/3 mL; p<0.001). Transcripts of ß-actin and ZO-1 were significantly upregulated in response to DON, irrespective of direction, whereas IL-8 mRNA remained unaffected. However, ZO-1 spatial distribution in the tight junction and its function (TEER) were detrimentally affected by basolateral DON only. In conclusion, direction of DON exposure affected IPEC-J2 differently on a translational and functional level, but was mainly inconsequential on a transcriptional level.

[Treatment of myofascial lumbar dorsal pain. Effective clinical diagnostics and therapy]. / Die Behandlung myofaszialer lumbaler Rückenschmerzen. Effektive Diagnostik und Therapie.

BACKROUND: Lumbar dorsal pain is a problem that must be taken seriously and is part of many people's everyday lives. Not only does it cause high costs for the health system, it also frequently leads to inability to work. The significance of the myofascial system is still not taken seriously enough in therapy and clinical diagnostics, when treating dorsal pain. In the following article, the effectiveness of specifically targeted therapy for myofascial pain is evaluated. METHODS: Included in the study were 44 patients referred for lumbar dorsal treatment-resistant pain to a practice specializing in pain therapy. Therapy focused on treatment of the affected muscle area with physiotherapy and the additional techniques of infiltrating trigger points, neural therapy, and general relaxation exercises. Medication was optimized according to the specific guidelines for the condition. The effect of therapy was evaluated using the German pain questionnaire (Deutsche Schmerzfragebogen). RESULTS: After the therapy phase, patients had a significantly lower intensity of pain, anxiety, and depression, as well as an increased quality of life. CONCLUSION: The results indicate that targeted treatment of muscles and fascia in patients with chronic back pain can lead to a reduction of pain symptoms. The consideration of the myofascial systems, particularly in relation to nonspecific back pain, could contribute to improving the treatment of pain and contribute to lowering costs.

Deoxynivalenol affects the composition of the basement membrane proteins and influences en route the migration of CD16(+) cells into the intestinal epithelium.

The numerous pores in the basement membrane (BM) of the intestinal villi are essential for the communication of enterocytes with cells in the lamina propria, an important mechanism for the induction of intestinal immune responses. The intestinal epithelial barrier is affected by the mycotoxin deoxynivalenol (DON) from both the apical (luminal) and basolateral (serosal) side. The pig is the most susceptible species to the anorectic and immune-modulating effects of DON, which is most prevalent in crops. We analysed in pigs the effect of DON-contaminated feed on the composition and perforation of the BM and the presence of CD16(+) cells or their dendrites in the epithelium. In addition to in vivo experiments, in vitro studies were carried out. Using microarray analyses, the effects of DON on IPEC-J2 cells were studied with the focus on the BM. Our in vivo results showed in the control pigs: (1) a significant increased pore number (p ≤ 0.001) in the jejunum in comparison to ileum, (2) no difference in the pore size, and (3) comparable frequency of intraepithelial CD16(+) cells/dendrites in the jejunum and ileum. There was a marked trend that DON feeding increases: (1) the pore number in jejunum, and (2) the number of CD16(+) cells/dendrites in the epithelium (Tukey-Kramer; p = 0.055 and p = 0.067, respectively). The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells). The down-regulation of genes like syndecan, fibulin 6 and BM-40 was observed. These proteins are important factors in the BM composition and in formation of pores. Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells. Thus, DON affects the composition of the BM.

Interactions between the Fusarium toxin deoxynivalenol and lipopolysaccharides on the in vivo protein synthesis of acute phase proteins, cytokines and metabolic activity of peripheral blood mononuclear cells in pigs.

The in vivo effects of the Fusarium toxin deoxynivalenol (DON) on albumin and fibrinogen synthesis in pigs and metabolic activity of porcine peripheral blood mononuclear cells (PBMCs) were studied alone or in combination with lipopolysaccharides (LPSs) in order to examine proposed synergistic effects of both substances. A total of 36 male castrated pigs (initial weight of 26 kg) were used. Uncontaminated (Control) and naturally DON-contaminated (chronic oral DON, 3.1mg/kg diet) wheat was fed for 37 days. On the day of protein synthesis measurement, pigs recruited from the Control group were treated once intravenously with (iv) DON (100 µg/kg live weight (LW)/h), iv LPS (7.5 µg/kgLW/h) or a combination of both substances, and six pigs from the chronic oral group were treated once with iv LPS. A treatment with DON alone exhibited no alterations of acute phase protein synthesis and metabolic activity of PBMC. There was no evidence that the chosen dosing regimen of DON had influences on the induced sub-acute stage of sepsis, as the LPS challenge, irrespective of DON co-exposure, mediated an acute phase reaction with a typical decrease of albumin synthesis, as well as changes in cytokine concentration and a loss of metabolic activity in PBMC.

Deoxynivalenol and E.coli lipopolysaccharide alter epithelial proliferation and spatial distribution of apical junction proteins along the small intestinal axis.

We investigated a proposed synergistic effect of deoxynivalenol (DON) and lipopolysaccharides (LPS) on small intestinal architecture and epithelial barrier integrity in pigs. Crypt depth and intestinal cell proliferation were analyzed, as well as expression of zonula occludens protein-1 (ZO-1) and ß-catenin of the apical junction complex along the small intestine. Barrows (26.2±4.1 kg) were fed restrictedly either a control diet (CON) or a diet naturally contaminated with 3.1 mg DON/kg feed (DON) for 37 d. At d 37, the control group was infused for 1 h either with 100 µg/kg BW of DON (CON-DON, n=6), 7.5 µg/kg BW of LPS (CON-LPS, n=6), a combination of both (CON-DON+LPS, n=7), or 0.9% NaCl (CON-CON, n=6) and the DON group with 7.5 µg/kg BW of LPS (DON-LPS, n=8) or 0.9% NaCl (DON-CON, n=6). Pigs were euthanized 3.25 h after start of infusion. Immunohistochemistry (5'-bromo-2'-deoxyuridine for proliferation) and immunofluorescence (ZO-1 and ß-catenin) from duodenum, proximal jejunum, mid-jejunum, proximal ileum, and terminal ileum were analyzed for crypt depth, cell proliferation, and apical junction proteins. Duodenal crypts were deeper compared with the other 4 intestinal regions, and proximal jejunal crypts were deeper than those of mid-jejunum and proximal ileum (P<0.001). Epithelial proliferation showed a bell-shaped distribution along the small intestinal axis. Duodenal proliferating cells had the least number compared with jejunal sections and proximal ileum (P<0.001). Neither DON nor LPS affected these variables. Zonula occludens-1 displayed a distinct spatial distribution in the epithelium with an apical and a cytosolic component. Apical expression of ZO-1 was severely damaged in the mid-jejunum (P<0.001) of CON-DON compared with animals treated with LPS. Also, in all animals receiving LPS systemically, the cytosolic ZO-1 fraction in the 3 upper gut sections disappeared completely. This effect was independent of DON presence. Control pigs had a greater basolateral ß-catenin accumulation (P<0.05) in the cells, whereas the protein distribution did not differ in CON-DON pigs. In conclusion, results of this experiment demonstrated that epithelial proliferation has a distinct pattern along the small intestine and is not necessarily positively linked to crypt depth in pigs. Furthermore, results indicate that LPS changed the spatial distribution of ZO-1. A synergistic effect of DON and LPS on intestinal architecture could not be verified in the present study.

Air-liquid interface cultures enhance the oxygen supply and trigger the structural and functional differentiation of intestinal porcine epithelial cells (IPEC).

The specific function of the epithelium as critical barrier between the intestinal lumen and the organism's internal microenvironment is reflected by permanent maintenance of intercellular junctions and cellular polarity. The intestinal epithelial cells are responsible for absorption of nutritional components, facing mechanical stress and a changing oxygen supplementation via blood stream. Oxygen itself can regulate the barrier and the absorptive function of the epithelium. Therefore, we compared the dish cell culture, the transwell-like membrane culture and the oxygen enriched air-liquid interface (ALI) culture. We demonstrated strong influence of the different culture conditions on morphology and function of intestinal porcine epithelial cell lines in vitro. ALI culture resulted in a significant increase in cell number, epithelial cell layer thickness and expression as well as apical localisation of the microvilli-associated protein villin. Remarkable similarities regarding the morphological parameters were observed between ALI cultures and intestinal epithelial cells in vivo. Furthermore, the functional analysis of protein uptake and degradation by the epithelial cells demonstrated the necessity of sufficient oxygen supply as achieved in ALI cultures. Our study is the first report providing marked evidence that optimised oxygen supply using ALI cultures directly affects the morphological differentiation and functional properties of intestinal epithelial cells in vitro.

Studies on the toxicity of deoxynivalenol (DON), sodium metabisulfite, DON-sulfonate (DONS) and de-epoxy-DON for porcine peripheral blood mononuclear cells and the Intestinal Porcine Epithelial Cell lines IPEC-1 and IPEC-J2, and on effects of DON and DONS on piglets.

The in vitro effects of deoxynivalenol (DON), de-epoxy-DON, DON-sulfonate (DONS) and sodium metabisulfite (Na(2)S(2)O(5), SBS) on porcine peripheral blood mononuclear cells (PBMC), and on the Intestinal Porcine Epithelial Cell lines IPEC-1 and IPEC-J2 were examined by using the MTT assay. In addition, an uncontaminated and a DON contaminated triticale were included in diets either untreated (CON, FUS) or SBS treated (CON-SBS, FUS-SBS) and fed to piglets for 28 d starting from weaning. The diet concentrations of DON and DONS amounted to 0.156, 2.312, 0.084 and 0.275 mg and to<0.05, <0.05, <0.05 and 1.841 mg/kg, respectively. PBMC of the so-exposed piglets were also subjected to the MTT assay. Neither DONS and SBS nor de-epoxy-DON affected the viability of PBMC, IPEC-1 and IPEC-J2 significantly up to concentrations of 17, 8 and 23 microM, respectively. For DON, IC(50) values were estimated at 1.2+/-0.1, 1.3+/-0.5 and 3.0+/-0.8 microM for PBMC, IPEC-1 and IPEC-J2, respectively. PBMC from piglets fed the SBS treated diets were characterized by a significantly decreased stimulation index and an increased IgA supernatant concentration with the SBS effect being significantly more pronounced after feeding the FUS-SBS diet. Further studies should clarify the possible impact of SBS on the porcine immune system.

Natural alternatives to in-feed antibiotics in pig production: can immunomodulators play a role?

As a result of the European ban of in-feed growth-promoting antibiotics, new strategies are being developed to increase the resistance to disease in farm animals. In pig production, this is of particular importance during the weaning transition when piglets are subjected to major stressful events, making them highly sensitive to digestive disorders. At this time, the development of both innate and adaptive immunity at the mucosal surface is critical in preventing the potential harmful effects of intestinal pathogenic agents. Strategies aiming at stimulating natural host defences through the use of substances able to modulate immune functions have gained increasing interest in animal research, and different bioactive components a priori sharing those properties have been the subject of in vivo nutritional investigations in pig. Among these, yeast derivates (ß-glucans and mannans) are able to interact with immune cells, particularly phagocytic cells. However, studies where they have been fed to pigs have shown inconsistent results, suggesting that their ability to target the sensitive immune cells through the oral route is questionable. The plant extracts, which would benefit from a positive image in the public opinion, have also been tested. However, due to a lack of data on the bioactive components of particular plants and the large diversity of species, it has proved difficult to prepare extracts of equivalent potency and thus, the literature on their influence on pig immunity remains inconclusive. In considering piglet immunity and health benefits, the most promising results to date have been obtained with spray-dried animal plasma, whose positive effects would be provided by specific antibodies and non-specific competition of some plasma components with bacteria for intestinal receptors. The major positive effect of spray-dried animal plasma is in reducing the infiltration of gut-associated lymphoid tissue by immune cells, which is likely to be the result of a decreased colonisation by potentially harmful bacteria. This review also highlights the limitations of some of the published in vivo studies on the immunomodulatory activity of certain feed additives. Among those, the lack of standardisation of extracts and the heterogeneity of piglet-rearing conditions (e.g. exposure to pathogens) are likely the most limiting.

Cholera toxin transiently inhibits porcine T cell proliferation in vitro.

Cholera toxin (Ctx) is an important mucosal adjuvant with potential experimental applications in pigs. However, little is known about the direct effects of Ctx on porcine immune cells. Therefore, we analysed the influence of Ctx on mitogen-induced lymphocyte proliferation. Ctx inhibited peripheral blood mononuclear cell (PBMC) proliferation with an IC50 of 34+/-17 ng/mL. This inhibition was not due to increased cell death. Lymphoblast formation in cultures stimulated with concanavalin A and Ctx was decreased at 24 h, but had reached the levels of control cultures again at 72 and 120 h, indicating that suppression was transient. Analysis of T cell subsets revealed that Ctx treatment specifically reduced the percentage of CD4-CD8+ and gammadelta T cells, whereas the proportion of CD4+CD8- increased. Furthermore, Ctx caused secretion of IL-10 by PBMC cultures, but depressed TNFalpha secretion.

Research note: Effects of deoxynivalenol on immunohistological parameters in pigs.

TheFusarium toxin deoxynivalenol (DON) is known to exert immunomodulatory effects. Numerous studies in mice demonstrated that dietary exposure leads to an upregulation of polymeric serum immunoglobulin A (IgA) suggesting the mucosal immune system as a primary target while at the cellular level T cells and macrophages are involved in this process.The present study aimed to verify these effects in pigs. A total of 24 male pigs were subjected to four treatments, a control group fed a diet devoid of DON, a chronically exposed group receiving a diet containing contaminated wheat (5.7 mg DON/kg diet), an acute orally exposed group receiving only one meal (550 g) of the contaminated feed and an acute intravenous exposed group receiving 53 µg DON/kg body weight. Cryosections of the spleen and the jejunum of the pigs were immunohistologically stained for IgA+, CD3+, CD4+ and CD8+ cells. The number of positive stained cells did not differ significantly between the treatment groups and the control group of any of the specimen.

On the effects of a chronic deoxynivalenol intoxication on performance, haematological and serum parameters of pigs when diets are offered either for ad libitum consumption or fed restrictively.

Wheat infected naturally with Fusarium, contaminated mainly with deoxynivalenol (DON) (16.6 mg DON/kg), was added to a total constant wheat content of 400 g/kg diet. To distinguish between differences in feed intake and specific effects of the DON contaminated diet, control and DON contaminated feed was administered for 11 weeks under ad libitum and restrictive feeding conditions to 48 pigs of both sexes, which were randomly divided into four groups (n = 12 per group). Feed intake was 2.90 kg/day, live weight gain 987 g/day and feed to gain ratio 2.77 kg/kg for the ad libitum fed control group. The group fed DON contaminated wheat ad libitum significantly consumed 15% less feed and gained 13% less live weight, while the feed to gain ratio was unaffected. Moreover, it was concluded that the lower growth performance by DON contaminated feed resulted mainly from the lower voluntary feed intake, because there were no differences in live weight gain between the groups with the restrictive feeding regimen. On the contrary, metabolizable energy, nitrogen retention digestibility of organic matter, crude protein, crude fat and crude fibre were significantly increased by 3, 10, 3, 6, 9 and 20% in the DON group respectively. Animals fed DON contaminated diets needed more time to consume the restrictive ration than the control group. For example in the first hour after feeding 85% of the control pigs had consumed all feed, but only 39% of the DON group had. There were only few differences in haematological and serum parameters, which were characterized by a high variation between individuals. DON and IgA concentrations in serum were significantly influenced by DON exposure.

The pig as a model of developmental immunology.

There are many limitations to analyse the developing immune system in humans, thus there is need for experimental animal models to study the environmental influences during the ontogeny of the immune system. However risk assessment is difficult in using rodent models alone, especially as the intrauterine period of development is much shorter than that of humans. In addition to studies in dogs, the pig provides a variety of experimental approaches for developmental immunotoxicology. The gestation period is 115 days and the occurrence of the different lines of T and B lymphocytes in the blood and organs of the porcine embryo and fetus is well documented. Fetal porcine B cells represent a naive population developing without maternal idiotypic-antiidiotypic influences. The postnatal development is highly correlated to sufficient uptake of colostrum during the first 48 hours. Although many immunotoxicological experiments have been performed, there is a limited number of original publications about these studies. With the different strains of standard pigs and miniature pigs available and the rapid growing amount of immunological reagents, the pig represents an important experimental model for cost-effective studies in developmental immunotoxicology to analyse the risk of environmental hazards.

Model for experimental revascularized laryngeal allotransplantation.

BACKGROUND: Although a human laryngeal transplant has been undertaken successfully, important questions remain that require a suitable animal model. METHODS: A pig model for allotransplantation has been developed. Organ perfusion was studied in nine animals before four transplants were performed in congenic (unrecovered) animals and eight in unmatched (recovered) animals. Larynges were regularly examined endoscopically until death at 14 days. Immunosuppression included the use of tacrolimus. Revascularization was achieved by anastomosing the donor right cervical vascular tree to the recipient common carotid. In recovered animals, four allografts were placed orthotopically and four heterotopically. RESULTS: The pig larynx was perfused adequately via the right cervical vascular tree and congenic grafts were well tolerated. Of eight allografts, seven were well tolerated and remained healthy for the duration of the study (14 days). One allograft became infected between days 4 and 7 after operation. Median operating time was 6 h, with a median cold ischaemia time of 3 h. CONCLUSION: Revascularized allotransplants of the larynx can be undertaken reliably in pigs and this provides a preclinical model for studies of laryngeal transplantation.

Actinobacillus pleuropneumoniae iron transport and urease activity: effects on bacterial virulence and host immune response.

Actinobacillus pleuropneumoniae, a porcine respiratory tract pathogen, has been shown to express transferrin-binding proteins and urease during infection. Both activities have been associated with virulence; however, their functional role for infection has not yet been elucidated. We used two isogenic A. pleuropneumoniae single mutants (DeltaexbB and DeltaureC) and a newly constructed A. pleuropneumoniae double (DeltaureC DeltaexbB) mutant in aerosol infection experiments. Neither the A. pleuropneumoniae DeltaexbB mutant nor the double DeltaureC DeltaexbB mutant was able to colonize sufficiently long to initiate a detectable humoral immune response. These results imply that the ability to utilize transferrin-bound iron is required for multiplication and persistence of A. pleuropneumoniae in the porcine respiratory tract. The A. pleuropneumoniae DeltaureC mutant and the parent strain both caused infections that were indistinguishable from one another in the acute phase of disease; however, 3 weeks postinfection the A. pleuropneumoniae DeltaureC mutant, in contrast to the parent strain, could not be isolated from healthy lung tissue. In addition, the local immune response-as assessed by fluorescence-activated cell sorter and enzyme-linked immunosorbent spot analyses-revealed a significantly higher number of A. pleuropneumoniae-specific B cells in the bronchoalveolar lavage fluid (BALF) of pigs infected with the A. pleuropneumoniae DeltaureC mutant than in the BALF of those infected with the parent strain. These results imply that A. pleuropneumoniae urease activity may cause sufficient impairment of the local immune response to slightly improve the persistence of the urease-positive A. pleuropneumoniae parent strain.