Effects of antioxidants on rapeseed oil oxidation in an artificial digestion model analyzed by UHPLC-ESI-MS

A normal diet contains large quantities of oxidized fatty acids, glycerolipids, cholesterol, and their cytotoxic degradation products because many foods in the diet are fried, heated, or otherwise processed and consumed often after long periods of storage. There is also evidence that the acid medium of the stomach promotes lipid peroxidation and that the gastrointestinal tract is a major site of antioxidant action, as demonstrated by various colorimetric methods. The identity and yields of specific products of lipid transformation have seldom been determined. The present study describes the molecular species profiles of all major gastrointestinal lipids formed during digestion of autoxidized rapeseed oil in an artificial digestion model in the presence of L-ascorbic acid, 6-palmitoyl-O-L-ascorbic acid, 3,5-di-tert-butyl-4-hydroxytoluene (BHT), DL-α-tocopherol, and DL-α-tocopheryl acetate. Differences in oxidized lipid profiles were detected in the samples digested in the presence of different antioxidants, but none of them could prevent the formation of oxidized lipids or promote their degradation in a gastric digestion model. The lack of effect is attributed to the inappropriate nature of the gastrointestinal medium for the antioxidant activity of these vitamins and BHT. A fast ultrahigh performance liquid chromatographic-electrospray ionization-mass spectrometric method was developed for the analysis of lipolysis products, including epoxy, hydroperoxy, and hydroxy fatty acids, and acylglycerols, utilizing lithium as ionization enhancer.     

Prevalence of antibodies against Leptospira sp. in snakes,lizards and turtles in Slovenia

Background

Leptospiral infections in poikilothermic (cold blooded) animals have received very little attention and the literature concerning natural infections of these animals is limited. The aim of this study was to determine the prevalence of leptospiral antibodies in reptiles, imported into Slovenia and intended to be pets in close contact with humans. A total of 297 reptiles (22 snakes, 210 lizards and 65 turtles) were tested for specific antibodies against serovars of Leptospira interrogans sensu stricto using the microscopic agglutination test (MAT). Live cultures of different serovars were used as antigens. MAT was performed according to standard procedures and the degree of reaction was interpreted by estimating the percentage of agglutinated leptospires. Samples showing titres of ≥ 50 against one or more serovars were considered as positive.

Results

Antibodies against seven pathogenic serovars of L. interrogans sensu stricto were detected in 46 of 297 reptiles. Among 22 snakes, specific antibodies against pathogenic serovars of three Leptospira species (L. interrogans, L. kirschneri and L. borgpetersenii) at titre levels from 1:50 to 1:400 were detected in 6 snakes. In 31 of 210 lizards, specific antibodies were found in titres from 1:50 to 1:1000 and, finally, among 65 turtles (terrapins and tortoises), 9 had specific antibodies at titre levels between 1:50 and 1:1600. Animals imported from non-EU countries showed significantly higher prevalence (25.0%; 95 confidence interval: 16.7–33.3%) than animals from EU member states (10.4%; confidence interval: 6.1–14.7%).

Conclusions

Reptiles may be considered as potential reservoirs of L. interrogans sensu stricto. Origin of the animals is a risk factor for presence of leptospiral antibodies, especially in lizards. Special attention should be focused on animals from non-EU member states.     

To jump or not to jump? Strategies employed by leisure and sport horses

Show jumping is one of the most popular equestrian disciplines although strategic jumping per se is seldom used by free-living horses when negotiating obstacles that they might otherwise avoid. To establish if horses are naturally motivated to jump, we examined horses in a free-choice situation when negotiating various obstacles under test conditions. Eighteen leisure horses (LHs) and 16 sport horses (SHs) participated in a number of 2-choice tests. First, horses were presented with 2 optional routes during a “free-choice test” to reach a food stimulus. The choices were a shorter route (over an obstacle) or a longer route (around the obstacle). During 8 consecutive trials, the horses encountered an increase in obstacle height on every second trial as follows: 0, 20, 35, and 50 cm. In the “ridden test” after an initial conditioning period, the horses were tested with a Y-maze formation during 3 consecutive trials, where one element of the maze contained an obstacle arm (OA). The horses walked or trotted over the obstacle most frequently (59.9%) and jumped the obstacle in only 10.7% of the cases. For both the LH and SH groups, the horses' motivation to traverse the obstacle decreased as the obstacle height increased and only 44.1% of horses actually negotiated the 50-cm obstacle as compared with the 20-cm obstacle (85.3%). The LH group preferred to go around the obstacle significantly more often than the SH group (24 trials, 16.7% vs. 9 trials, 7.0%, for SH and LH, respectively; χ² = 6.81, N = 33, P = 0.0090). The LH group used a jumping strategy to clear the obstacle far less frequently (6 trials, 4.2%) than the SH group (23 trials, 18.0%; χ² = 9.96, N = 29, P = 0.0023). In the Y-maze trials, the LH group exhibited preference (40 per 54 trials, 74.1%; χ² = 12.5, P < 0.0001), and the SH group failed to exhibit any preference (20 per 48 trials; χ² = 1.33, P = 0.3123). There was no evidence of any correlation between the motivation to clear the obstacle and total number of OA choices in either the LH group (r_s = 0.13, P = 0.9594) or the SH group (r_s = ?0.25, P = 0.3492). The findings from the present study indicate that SHs are motivated and willing to jump obstacles more often than are LHs under similar conditions. However, the apparent reluctance of the horses overall to continue jumping as the obstacle height increases suggests that, in general, many horses could easily encounter excessive demands (overfacing) in sport. This issue should be carefully monitored in terms of equine training, competition, and welfare.     

Immune defense of wild‐caught Norway rats is characterized by increased levels of basal activity but reduced capability to respond to further immune stimulation

Studies of wild animals' immunity often use comparison with laboratory‐raised individuals. Using such an approach, various data were obtained concerning wild Norway rat's immunity. Lower or higher potential of immune system cells to respond to activation stimuli were shown, because of analysis of disparate parameters and/ or small number of analyzed individuals. Inconsistent differences between laboratory and wild rats were shown too, owing to great response variability in wild rats. We hypothesized that wild rats will express more intense immune activity compared to their laboratory counterparts which live in a less demanding environment. To test this, we analyzed the circulating levels of inflammatory cytokine interleukin‐6 (IL‐6), a mediator which has a central role in host immune defense. In addition, we examined the activity of the central immune organ, the spleen, including cell proliferation and production of pro‐inflammatory cytokines interferon‐γ (IFN‐γ) and interleukin‐17 (IL‐17), which are major effectors of cellular adaptive immune response. In order to obtain reasonable insight into the immunity of wild Norway rats, analysis was conducted on a much larger number of individuals compared to other studies. Higher levels of plasma IL‐6, higher spleen mass, cellularity and basal IFN‐γ production concomitantly with lower basal production of anti‐inflammatory cytokine interleukin‐10 (IL‐10) revealed more intense immune activity in the wild compared to laboratory rats. However, lower responsiveness of their spleen cells' proinflammatory cytokine production to concanavalin A (ConA) stimulation, along with preserved capacity of IL‐10 response, might be perceived as an indication of wild rats' reduced capability to cope with incoming environmental stimuli, but also as a means to limit tissue damage.     

Skin biopsies predict acute graft-versus-host disease after small bowel transplantation in pigs

Intestinal transplantation is being increasingly performed to treat patients with irreversible intestinal failure. The major cause of intestinal graft failure is graft-versus-host disease (GVHD) that represents a life-threatening complication after small bowel transplantation (Itx). The purpose of this study was to assess the diagnostic and prognostic value of skin biopsy histological changes for acute GVHD after Itx in pigs. Thirty-four Large White pigs were divided into three groups: Group 1 with Itx only, Group 2 with Itx and donor bone marrow infusion (Itx BM) and Group 3 (control group - before the operation). Animals received tacrolimus-based immunosuppression from day 0 to day 30 postoperatively. Skin and small bowel biopsies were histologically assessed, analysed and classified from grade 1 to 4 on postoperative days 15, 30, 45 and 60. There was a strong correlation between the histological grading values of skin biopsy changes and the histological grading values of small bowel biopsy changes (Kendall's tau_b is 0.855 for the Itx group and 0.730 for the Itx BM group). The significant correlation found between skin and small bowel histological changes suggests the prognostic value of skin biopsies after Itx. In conclusion, our findings emphasise the diagnostic and prognostic value of skin biopsy analysis for acute GVHD after Itx.