Štefan Faix

Effects of deoxynivalenol and zearalenone on oxidative stress and blood phagocytic activity in broilers

Effects of dietary contamination with various levels of deoxynivalenol (DON) and zearalenone (ZEA) were investigated on Ross 308 hybrid broilers of both sexes. After hatching, all chickens were fed an identical control diet for two weeks. Then chickens of Group 1 received a diet contaminated with DON and ZEA, both being 3.4 mg · kg−1, while Group 2 received DON and ZEA at 8.2 and 8.3 mg · kg−1, respectively. The diet of the control group contained background levels of mycotoxins. Samples of blood and tissues were collected after two weeks. Intake of both contaminated diets resulted in a significantly decreased activity of glutathione peroxidase (GPx) and increased level of malondialdehyde (MDA) in liver tissue, while in kidneys the concentration of MDA was significantly increased only in Group 1. On the other hand, activities of blood GPx and plasma γ-glutamyltransferase (GGT) were elevated in Group 2 only. Activities of thioredoxin reductase in liver and GPx in duodenal mucosa tissues, superoxide dismutase (SOD) in erythrocytes as well as levels of MDA in duodenal mucosa and α-tocopherol in plasma were not affected by dietary mycotoxins. Blood phagocytic activity was significantly depressed in Group 1 and 2. These results demonstrate that diets contaminated with DON and ZEA at medium levels are already able to induce oxidative stress and compromise the blood phagocytic activity in fattening chickens.

Effects of excessive selenium supplementation to diet contaminated with deoxynivalenol on blood phagocytic activity and antioxidative status of broilers.

This study examined the effects of excessive dietary supplementation with organic selenium on phagocytic activity and antioxidative status of chickens for fattening fed diet contaminated with deoxynivalenol (DON). Sixty chickens of Ross 308 hybrids were at day of hatching divided into four groups with 15 birds in each. The background DON dietary levels in both negative and positive control groups were 0.2 mg/kg. The complete feed for positive control group was supplemented with Se dose 1 mg/kg in the form of Se-yeast. Group 3 was fed diet with DON level 3 mg/kg while diet for group 4 combined DON level 3 mg/kg with a excessive supplement of Se-yeast (Se dose 1 mg/kg). After 6 weeks of dietary intake, six randomly-chosen chickens from each group were sampled. Feeding of contaminated diet resulted in significantly reduced blood phagocytic activity (19.5 ± 1.1% in the negative control vs. 12.8 ± 0.8% in the DON-treated group, p < 0.05). Se-yeast supplemented to the DON contaminated diet prevented suppression of phagocytic activity. Dietary intake of DON at levels 3 mg/kg did not influence the plasma α-tocopherol level while excessive dietary Se dose reduced it in both Se supplemented groups. Neither the birds of DON-treated group nor the birds from group 4 with DON and Se-yeast showed any response in plasma γ-glutamyltransferase (GGT) activity. Subtoxic dietary level of DON significantly increased the activity of glutathione peroxidase (GPx) in the duodenal mucosa, while additional Se supplementation prevented such a response to mycotoxin. On the other hand, both Se supplemented groups showed significantly elevated GPx activities in blood, liver and kidney, (p < 0.05). The results suggest a potential ability of excessive supplementation of organic selenium to prevent the blood phagocytic activity suppression and changes in GPx activity in duodenal tissue induced in broilers by subtoxic dietary levels of DON.

The Effects of Organic Selenium Supplementation on the Rumen Ciliate Population in Sheep

The effect of selenium supplementation on the rumen protozoan population of sheep was demonstrated. Both the total and generic counts of rumen ciliates in sheep fed a diet with basal Se content (70 µg/kg dry matter) were compared to those of animals given feed supplemented with inorganic (disodium selenite) or organic Se (selenized yeast) (310 µg/kg dry matter). The genera ofEntodinium, Isotricha, Dasytricha, Ophryoscolex, Diploplastron andPolyplastron occurred in all sheep except for the control, in whichOphryoscolex was not observed. The population ofOphryoscolex caudatus f.tricoronatus was significantly higher in sheep supplemented with organic Se than in animals given inorganic Se (by 160 %). Supplementation of feed with selenized yeast induced significant growth in theDiploplastron population (by 63 %) while no change occurred in sheep given selenite. The populations ofDasytricha ruminantium andPolyplastron multivesiculatum were higher than control in both Se-supplemented groups. The ciliate population ofEntodinium spp. was not influenced by Se supplements. Our results suggest a protective effect of Se feed supplementation on the development of some rumen ciliate species in young ruminants.

Effect of Enterococcus faecium AL41 and Thymus vulgaris essential oil on small intestine integrity and antioxidative status of laying hens.

We investigated the effect of Enterococcus faecium on phagocytic activity, antioxidative status in vivo and the effect of E. faecium and 0.4% concentration of Thymus vulgaris essential oil (EO) on the duodenal tissue integrity in vitro in laying hens. The birds were fed the same standard diets and were divided into four groups. E. faecium was added to the drinking water for the second and fourth groups. EO was added to special chambers for measuring trans-epithelial electrical resistance (TEER) for the third and fourth groups only. TEER was lower in groups where EO was added, but in the group with E. faecium TEER was not changed significantly. Our results show that EO at 0.4% concentration may negatively affect intestine integrity, and the probiotic strain E. faecium AL41 is able to eliminate this effect and can strengthen non-specific immunity. To confirm our findings further histopathological investigations of intestinal tissue are needed.

Effect of thyme oil on small intestine integrity and antioxidant status, phagocytic activity and gastrointestinal microbiota in rabbits.

The effects of 0.5 g thyme oil per kg dry matter (DM) of diet on duodenal tissue integrity, antioxidant status, phagocytic activity and selected microbiota in the caecum and faeces of rabbits were studied. Twenty-four rabbits were divided into two groups and were fed a commercial granulated diet for growing rabbits (CD) with access to water ad libitum. The first group was fed the CD, while to the CD of the second group thyme oil was added. Intestinal integrity was tested by measuring the transepithelial electrical resistance (TEER). Thyme oil significantly increased the value of total antioxidant status (TAS) in the blood plasma and glutathione peroxidase (GPx) activity in the liver, and it decreased malondialdehyde (MDA) concentration in the duodenal tissue. Thyme oil resulted in strengthened intestinal integrity, as the essential oil supplementation significantly increased TEER values in the experiment. The faecal microbiota of rabbits was almost completely balanced in both groups, and only a slight decrease was found in the microbial population at day 42 of the trial. In both groups, the bacterial counts were generally lower in the caecum than in the faecal samples. In conclusion, dietary supplementation with 0.5 g/kg DM thyme oil may improve intestinal integrity, and it may have an antioxidant effect. A tendency was also found for thyme oil to stimulate the abundance of some microbes beneficial in the rabbit gut.

Effect of lignin on oxidative stress in chickens fed a diet contaminated with zearalenone

The effect of lignin supplementation to a diet contaminated with zearalenone (ZEA) on antioxidant status was studied in female chickens of the ISA BROWN laying strain. From the day of hatching to 2 weeks of age, four groups of chickens were fed the same uncontaminated control diet. After 14 days, Group 1 (control) continued to receive the uncontaminated diet, while Group 2 was fed an identical diet enriched with 0.5% chemically modified lignin. Simultaneously, chickens of Group 3 were switched to a diet contaminated with 7.9 mg/kg ZEA and those of Group 4 to an identical contaminated diet supplemented with 0.5% lignin. At 6 weeks of age blood and tissue samples were collected. Feeding of a diet contaminated with a high level of ZEA resulted in elevated glutathione peroxidase (GPx) activity in the duodenal mucosa and kidney tissues, and an increased γ-glutamyltransferase (GGT) activity in the plasma, indicative of oxidative stress. In the liver tissue, no mycotoxin-induced response in GPx and thioredoxin reductase (TrxR) activities occurred, and the malondialdehyde (MDA) level was even reduced. Neither the plasma levels of retinol and α-tocopherol nor the activities of superoxide dismutase in erythrocytes and GPx in blood were affected in birds fed the contaminated diet. The only effect of lignin supplemented to the contaminated feed was that it prevented the increase of GPx activity in the duodenal mucosa as an indicator of oxidative stress.

Effects of feeding diets contaminated with Fusarium mycotoxins on blood biochemical parameters of broiler chickens.

This study was conducted to investigate the effects of deoxynivalenol (DON) and zearalenone (ZEA) on some biochemical indices of broiler chickens. Twenty-four Ross 308 hybrid broiler chickens of both sexes were fed diets containing maize contaminated with Fusarium mycotoxins. The diets included a control diet (DON 0.60 mg/kg feed; ZEA 0.07 mg/kg feed), an experimental 1 diet (DON 3.4 mg kg⁻¹ feed; ZEA 3.4 mg kg⁻¹ feed), and an experimental 2 diet (DON 8.2 mg kg⁻¹ feed; ZEA 8.3 mg kg⁻¹ feed). Contaminated diets were fed from 14 days of age for 14 days. Blood samples were collected from 4-week-old birds. Chicks fed a diet containing a low level of contaminated maize (experimental 1) had decreased plasma potassium, magnesium, phosphorus, total protein, albumin, triglycerides, free glycerol concentrations and increased cholesterol and calcium levels as well as alkaline phosphatase (ALP) and aspartate aminotransferase (AST) enzyme activities as compared to the control. Feeding a diet contaminated with high levels of mycotoxins (experimental 2) resulted in decreased plasma potassium, magnesium, total protein, albumin, triglycerides, free glycerol concentrations and increased plasma ALP, alanine aminotransferase (ALT) and AST enzyme activities. The effect of mycotoxin-contaminated diets on ALP activity was dose dependent. Chloride concentration was not affected by the diets. It can be concluded that feeding diets contaminated with both levels of Fusarium mycotoxins significantly affected protein, lipid and mineral metabolism as well as AST and ALP enzyme activities in broiler chickens.

Identification and quantification of thymol metabolites in plasma, liver, and duodenal wall of broiler chickens using UHPLC‐ESI‐QTOF‐MS

In the present study, we aimed to develop a method for thymol sulfate and thymol glucuronide determination in plasma, liver and duodenal wall of broiler chickens after feeding with a Thymus vulgaris essential oil at the different concentrations (0.01, 0.05 and 0.1% w/w). UHPLC coupled with accurate-mass QTOF-MS was used for identification and quantification of thymol metabolites. Novel Waters Oasis Prime HLB solid-phase extraction cartridges were applied to sample clean-up with extraction recoveries ranged from 85 to 92%. The presence of thymol glucuronide was confirmed by MS software according to molecular formula, score, mass error and double bond equivalent. In terms of validation, calibration curves of thymol sulfate were constructed in matrix samples with linearity from 3.91 to 250.0 ng/mL and correlation coefficients were within the range of 0.9979-0.9995. Limits of detection were 0.97, 0.29 and 0.63 ng/mL and limits of quantification were 3.23, 0.97 and 2.09 ng/mL for plasma, liver and duodenal wall, respectively. Intra-day and inter-day precision expressed as relative standard deviation were <4.35%. To highlight, thymol metabolites were directly detected for the first time in liver and duodenal wall and this method was shown to be successfully applicable for investigation of thymol metabolism in chickens after thyme essential oil ingestion.

The anti-translocation and anti-inflammatory effect of cinnamon oil in mice with TNBS induced colitis

The bacterial translocation induced by colitis may cause the organ failure and sepsis. Therefore, it is necessary to find new possibilities for prevention and therapy of this problem. The purpose of this study was to examine Escherichia coli anti-translocation activity of cinnamon oil and its ability to reduce colonic damage in mice with TNBS (2,4,6-trinitrobenzenesulfonic acid) induced colitis. Mice received cinnamon essential oil in four various concentrations (0.5%, 0.25%, 0.125% and 0.063%) in the powdery commercial rodent diet, starting 21 days before induction of TNBS colitis. The colonic damage was assessed using the colon macroscopic scoring system (Wallace score). E. coli translocation to the mesenteric lymphatic nodules was evaluated by serial dilutions method for counting bacteria. Bacterial translocation was significantly reduced in first and third group (15.2% or 42.8% in cinnamon oil groups versus 100% in TNBS group). Cinnamon oil was effective also against the colonic damage in all cinnamon oil groups (macroscopically scores of grade 9 in TNBS group versus 5.25, 5.63, 5.13 and 3.25 in cinnamon oil groups). Our results confirmed that dietary administration of cinnamon oil could possess potential therapeutic effects on bacterial translocation and intestinal wall injury in colitis.

Effects of borneol on blood chemistry changes in chickens.

It is well established that borneol may be alter bone metabolism. The aim of the study was to investigate the effects of feeding diets supplemented with borneol on blood biochemistry of chickens. Forty chicks of a commercial strain ISA Brown were divided into five groups with 8 birds in each and fed diets supplemented with 0%, 0.1%, 0.05% or 0.025% borneol dissolved in sunflower oil from hatching up to week 11. The concentrations of potassium and magnesium were found to be significantly higher in chickens fed the diet supplemented with 0.1% and 0.025% borneol, respectively. The blood plasma levels of total protein and glucose in chickens treated with 0.05% borneol were significantly lowered. Feeding of diets supplemented with borneol significantly increased levels of bilirubin, cholesterol and total lipids in blood plasma compared to the control group supplemented with sunflower oil only. The present study shows that addition of borneol to the diets can influence some parameters of mineral, lipid and protein metabolism in chicken blood plasma.