T-2 toxin induced Salmonella Typhimurium intoxication results in decreased Salmonella numbers in the cecum contents of pigs, despite marked effects on Salmonella-host cell interactions

ABSTRACT: The mycotoxin T-2 toxin and Salmonella Typhimurium infections pose a significant threat to human and animal health. Interactions between both agents may result in a different outcome of the infection. Therefore, the aim of the presented study was to investigate the effects of low and relevant concentrations of T-2 toxin on the course of a Salmonella Typhimurium infection in pigs. We showed that the presence of 15 and 83 μg T-2 toxin per kg feed significantly decreased the amount of Salmonella Typhimurium bacteria present in the cecum contents, and a tendency to a reduced colonization of the jejunum, ileum, cecum, colon and colon contents was noticed. In vitro, proteomic analysis of porcine enterocytes revealed that a very low concentration of T-2 toxin (5 ng/mL) affects the protein expression of mitochondrial, endoplasmatic reticulum and cytoskeleton associated proteins, proteins involved in protein synthesis and folding, RNA synthesis, mitogen-activated protein kinase signaling and regulatory processes. Similarly low concentrations (1-100 ng/mL) promoted the susceptibility of porcine macrophages and intestinal epithelial cells to Salmonella Typhimurium invasion, in a SPI-1 independent manner. Furthermore, T-2 toxin (1-5 ng/mL) promoted the translocation of Salmonella Typhimurium over an intestinal porcine epithelial cell monolayer. Although these findings may seem in favour of Salmonella Typhimurium, microarray analysis showed that T-2 toxin (5 ng/mL) causes an intoxication of Salmonella Typhimurium, represented by a reduced motility and a downregulation of metabolic and Salmonella Pathogenicity Island 1 genes. This study demonstrates marked interactions of T-2 toxin with Salmonella Typhimurium pathogenesis, resulting in bacterial intoxication.     

Quantitative assessment of physiological and behavioural parameters in healthy dairy cows evoked by transcutaneous electrical nerve stimulation of the udder

The pain and distress associated with transcutaneous electrical nerve stimulation (TENS) of the udder was evaluated by treating 20 healthy dairy cows with an electrical udder stimulator. This generated a sequence of pulses (frequency: 160±10% impulses per second, duration 250 μs) and provided voltage ranges from 0 to 10 volts (±10%). Trials took place on three consecutive days, twice daily after morning and evening milking. Daily sessions were divided into two periods: (1) control (sham treatment) and (2) treatment (real treatment). Physiological (heart rate, respiratory rate, and plasma cortisol concentration) as well as ethological parameters (kicking, weight shifting, and looking backwards to udder) were defined as pain-indicating parameters and observed. Evaluation of data showed that only one parameter (kicking) was significantly increased during real treatment compared to sham treatment. It is concluded that the TENS therapy tested in this study can evoke changes in behaviour (increased kicking) consistent with an experience of pain in some cows.     

Influence of soil aeration on rooting and growth of the Beuys-trees in Kassel,Germany

Soil aeration is an important factor in tree growth. Oxygen must be taken from the atmosphere for root respiration, and carbon dioxide must be discharged to the atmosphere. Because the pore space of the soil could be considered the “dead end” of the free atmosphere, topsoil gas diffusivity is particularly important for soil aeration. Due to diverse land uses, several soil cover types alternate on a small scale at urban sites, competing with the natural function of soil as the living space for roots.During Documenta 7 in 1982, the artist Joseph Beuys initiated the spectacular landscape art project “7000 Oaks”. Seven thousand trees of approximately the same age were planted over the whole city of Kassel, Germany, offering best possible conditions for investigating the influence of specific site factors on root and tree development. At 8 different sites featuring 36 Beuys-oaks and 15 Beuys-planes, topsoil gas diffusivity, soil CO₂ concentration and soil respiration of different soil cover types were measured and correlated with fine root density and tree growth.Topsoil gas diffusivity and soil respiration depend on soil cover type. The lowest gas diffusivities and respiration rates were found at sealed sites, and the highest values were measured at vegetated sites such as lawn or flower beds. Soil gas diffusivity primarily controls soil respiration. Soil CO₂ concentration is not strictly linked to the coverage type and does not show a strictly directed dependence on top soil gas diffusivity and soil respiration. Tree root density and height as well as diameter at breast height (1.3 m) of the oaks were decisively shaped by the gas diffusivity of the soil cover, whereas the investigated planes were not affected by soil aeration deficiencies. The vitality of urban trees can be controlled by the design of the tree site and the choice of the species.     

Diversity declines in Microstegium vimineum (Japanese stiltgrass) patches

The spread of invasive plant species and their impacts on plant communities have received international attention as global trade and global environmental change enhance the colonization and establishment of introduced species and threaten the integrity of native ecosystems. Because introduced species vary in their impact, studying the relationship between invasion and native communities is necessary to guide allocation of finite management resources. By studying adjacent pairs of invaded and uninvaded plots across an eastern United States forested landscape, we demonstrate Microstegium vimineum was associated with local declines in species richness and cover of native species. Negative impacts of M. vimineum on species richness did not emerge until August when M. vimineum cover and height were greatest, highlighting the value of following study subjects through the growing season. In contrast, native species cover was already lower in invaded plots early in the growing season. M. vimineum invasion was not the only important driver of species richness and community composition within the study region; abiotic environmental gradients, such as soil nitrate concentration and pH, across the six study sites were also important in affecting species richness and cover, but lessened in explanatory power through the growing season. We conclude that M. vimineum has effects on community structure that may have long-term consequences for biodiversity. Studies which track sites through time and consider multiple scales are required as invaders impact multiple biotic and abiotic factors operating at different spatial and temporal scales.     

Nitrous oxide emissions after incorporation of winter oilseed rape (Brassica napus L.) residues under two different tillage treatments

The aim of this study was to investigate the effect of crop residues from winter oilseed rape on N₂O emissions from a loamy soil and to determine the effect of different tillage practices on N₂O fluxes. We therefore conducted a field experiment in which crop residues of winter oilseed rape (Brassica napus L., OSR) were replaced with ¹⁵N labelled OSR residues. Nitrous oxide (N₂O) emissions and ¹⁵N abundance in the N₂O were determined for a period of 11 months after harvest of OSR and in the succeeding crop winter wheat (Triticum aestivum L.) cultivated on a Haplic Luvisol in South Germany. Measurements were carried out with the closed chamber method in a treatment with conventional tillage (CT) and in a treatment with reduced soil tillage (RT). In both tillage treatments we also determined N₂O fluxes in control plots where we completely removed the crop residues. High N₂O fluxes occurred in a short period just after OSR residue replacement in fall and after N‐fertilization to winter wheat in the following spring. Although N₂O emissions differed for distinct treatments and sub‐periods, cumulative N₂O emissions over the whole investigation period (299 days) ranged between 1.7 kg and 2.4 kg N₂O‐N ha^?1 with no significant treatment effects. More than half of the cumulative emissions occurred during the first eight weeks after OSR replacement, highlighting the importance of this post‐harvest period for annual N₂O budgets of OSR. The contribution of residue N to the N₂O emission was low and explained by the high C/N‐ratio fostering immobilization of mineral N. In total only 0.03% of the N₂O‐N emitted in the conventional tillage treatment and 0.06% in the reduced tillage treatment stemmed directly from the crop residues. The ¹⁵N recovery in the treatments with crop residues was 62.8% (CT) and 75.1% (RT) with more than 97% of the recovered ¹⁵N in the top soil. Despite our measurements did not cover an entire year, the low contribution of the OSR residues to the direct N₂O emissions shows, that the current IPCC tier 1 approach, which assumes an EF of 1%, strongly overestimated direct emissions from OSR crop residues. Furthermore, we could not observe any relationship between tillage and crop residues on N₂O emission, only during the winter period were N₂O emissions from reduced tillage significantly higher compared to conventional tillage. Annual N₂O emission from RT and CT did not differ.     

Typing of Salmonella by DNA-microarrays

Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such as oligonucleotides or PCR products are immobilized in a high density, and compactness. Homologue DNA hybridises with the immobilized complementary nucleic acid probes. This study gives after a short general introduction in the principle of DNA-microarrays an overview about published data on the field of typing of Salmonella by microarrays. An onset of a DNA-microarray developed by the National Reference Laboratory for Salmonella (NRL-Salm) will be introduced. By this new technique, it is possible to answer epidemiological questions as well as to find genes involved in certain biochemical processes, such as pathogenicity or resistance of salmonellae.     

The role of nitric oxide in Listeria encephalitis of ruminants and in rats intracisternally infected with Listeria monocytogenes

Listeria monocytogenes is a Gram-positive facultative intracellular bacteria which infects a wide range of hosts. In ruminants, infection with L. monocytogenes frequently causes encephalitis, which is usually fatal in sheep and goat, while cattle often recover with antibiotic therapy. Since the role of NO in the control of Listeria is controversial, we have studied the expression of iNOS in the brains of cattle, sheep and goats which had succumbed to listeria encephalitis. iNOS was demonstrated in decreasing intensity in the M phi of microabscesses from cattle, sheep and goat. iNOS expression was accompanied by NT in the microabscesses of cattle, but was only present to a low degree in sheep and was absent in goats. This is indirect evidence for differences in the ability to produce NO in the three species. Presence of iNOS and NT were inversely correlated with the numbers of bacteria. While microabscesses of goats contained high amounts of L. monocytogenes they occurred only rarely in cattle. To corroborate our hypothesis that NO is involved in the control of listeria encephalitis a new animal model was developed. Eleven day old infant rats were infected intracisternally with a low dose of L. monocytogenes. This resulted in a transient meningoencephalitis with moderate clinical signs and low mortality. Listeria proliferated strongly in the inflammatory lesions during the first days of infection, reached a peak at day 4 and were eliminated until day 7. The presence of bacteria was closely accompanied by high numbers of iNOS-expressing M phi and the formation of NT. Administration of the iNOS inhibitor L-NIL or the radical scavenger PBN resulted in rapid death of the treated animals. However, the increase in bacterial numbers was one order of magnitude higher for animals treated with PBN compared with L-NIL administration. This shows that NO plays an important role in the control of a brain infection with Listeria, but suggests that reactive oxidants other than NO are also involved. In conclusion, our findings point to a possible involvement of the differences in the ability to express iNOS and subsequent NO production in the different clinical outcome of listeria encephalitis in cattle and small ruminants.     

Safety and accuracy of blind vs. ultrasound-guided dorsal retrobulbar nerve blocks in horses—A cadaveric study

Objective

There is limited knowledge regarding the safety and accuracy of ultrasound-guided retrobulbar nerve blocks in horses. The aim of this study was to compare these parameters between blind and ultrasound-guided injection techniques for the dorsal retrobulbar nerve block in horses.

Methods

Equine cadaver heads were used to inject the retrobulbar space with contrast medium (CM). Injections were performed either blindly based on anatomic landmarks (blind group, n = 44) or under ultrasonographic guidance (US-group, n = 44), equally divided between an experienced and unexperienced operator. Needle position and distribution of CM were assessed with computed tomography imaging and evaluated by a board-certified veterinary diagnostic imager blinded to the technique. Safety and accuracy of both techniques were compared.

Results

Ocular penetration was observed in two cases (n = 2/44) in the blind group but not in the US group (n = 0/44). No intrathecal, intraneural, or intravascular injections were seen in either group. Safety was significantly improved in the US group (p = .026). There was no statistically significant difference between the groups regarding the accuracy of the injection. Excellent accuracy was achieved more often with the ultrasound-guided technique (n = 11/22) than with the blind technique (n = 7/22) when performed by the unexperienced operator, but this difference was not statistically significant.

Conclusion

To prevent globe-threatening complications and improve the safety of the injection, we recommend using the ultrasound-guided injection technique for the dorsal retrobulbar nerve block.