The role of selenium and vitamin E in a Transylvanian enzootic equine recurrent rhabdomyolysis syndrome.

A severe form of recurrent exertional rhabdomyolysis occurs enzootically in a well-defined region of Transylvania, Harghita county. At the highest lying two settlements (more than 800 m above sea level), the prevalence of equine rhabdomyolysis is between 17 and 23%, while in the neighbouring villages in the valley it is less than 2%. The objective of our study was to clarify the role of selenium and vitamin E in the high prevalence of rhabdomyolysis in that region. Soil and hay samples were collected from each area to evaluate mineral content. Ten horses from the non-affected and 20 horses from the affected area were tested for serum selenium, vitamin E, glutathione peroxidase (GSH-Px), muscle enzymes, lactate and electrolytes. Hay samples collected from the affected area had lower selenium content. Horses in the affected regions had significantly lower serum selenium (P = 0.006) and GSH-Px levels than animals living in the non-affected regions. A good correlation between erythrocyte GSH-Px and serum selenium concentration could be demonstrated (r = 0.777, P < 0.001). Serum vitamin E levels were low independently of the origin of the horse. Based on our results, selenium deficiency possibly has a role in the Transylvanian enzootic equine recurrent rhabdomyolysis syndrome.

Fructose, glucose and fat interrelationships with metabolic pathway regulation and effects on the gut microbiota.

The purpose of this 30-day feeding study was to elucidate the changes, correlations, and mechanisms caused by the replacement of the starch content of the AIN-93G diet (St) with glucose (G), fructose (F) or lard (L) in body and organ weights, metabolic changes and caecal microbiota composition in rats (Wistar, SPF). The body weight gain of rats on the F diet was 12% less (P = 0.12) than in the St group. Rats on the L diet consumed 18.6% less feed, 31% more energy and gained 58.4% more than the animals on the St diet, indicating that, in addition to higher energy intake, better feed utilisation is a key factor in the obesogenic effect of diets of high nutrient and energy density. The G, F and L diets significantly increased the lipid content of the liver (St: 7.01 ± 1.48; G: 14.53 ± 8.77; F: 16.73 ± 8.77; L: 19.86 ± 4.92% of DM), suggesting that lipid accumulation in the liver is not a fructose-specific process. Relative to the St control, specific glucose effects were the decreasing serum glucagon (-41%) concentrations and glucagon/leptin ratio and the increasing serum leptin concentrations (+26%); specific fructose effects were the increased weights of the kidney, spleen, epididymal fat and the decreased weight of retroperitoneal fat and the lower immune response, as well as the increased insulin (+26%), glucagon (+26%) and decreased leptin (-25%) levels. This suggests a mild insulin resistance and catabolic metabolism in F rats. Specific lard effects were the decreased insulin (-9.14%) and increased glucagon (+40.44%) and leptin (+44.92%) levels. Relative to St, all diets increased the operational taxonomic units of the phylum Bacteroidetes. G and L decreased, while F increased the proportion of Firmicutes. F and L diets decreased the proportions of Actinobacteria, Proteobacteria and Verrucomicrobia. Correlation and centrality analyses were conducted to ascertain the positive and negative correlations and relative weights of the 32 parameters studied in the metabolic network. These correlations and the underlying potential mechanisms are discussed.

Effect of fulvic and humic acids on copper and zinc homeostasis in rats.

The objective of this study was to investigate the effects of fulvic acid (FA) and humic acid (HA), the two main compounds of humic substances (HSs), on copper (Cu) and zinc (Zn) homeostasis. Seventy-two male Wistar rats were randomly divided into nine experimental groups. The control diet (AIN-93G formula) and the diets supplemented with 0.1%, 0.2%, 0.4% and 0.8% FA or HA were fed for 26 days. Cu and Zn concentrations of the large intestinal content (LIC), liver, kidney, femur and hair were determined. FA and HA did not influence significantly the Cu or Zn contents of the experimental diets, the rats' feed intake, weight gain and the feed to gain ratio. Both FA and HA decreased the Cu concentrations of the LIC significantly and in a dose-related manner; however the absorption-stimulating effect of HA was more pronounced. FA increased the Cu content of the liver, but neither FA nor HA had a dose-dependent effect on it. FA or HA supplementations had no significant effect on the Cu concentration of the kidney. At the concentrations used, dietary FA or HA supplementations are not promising growth promoters. FA influences the Cu homeostasis unlike HA, because FA not only stimulates Cu absorption, but the extra quantity of absorbed Cu is retained in the organism. The stimulatory effect of HA on Zn absorption may not be manifested in Cu and Zn homeostasis, because of the tight connection of these microelements to FA and HA, which prevents the transmission of Zn from the ZnHA complex to the organs. As regards the effect of FA and HA on Cu and Zn homeostasis, both FA and HA stimulated the absorption of these microelements, but only FA increased the retention of Cu (in the liver) and Zn (in the kidney).

Effect of fulvic and humic acids on iron and manganese homeostasis in rats.

The objective of this study was to investigate the effects of fulvic acid (FA) and humic acid (HA) as the two main compounds of humic substances, separately on Fe and Mn homeostasis. Seventy-two male Wistar rats were randomly divided into 9 experimental groups. The control diet (AIN-93G formula) and diets supplemented with 0.1%, 0.2%, 0.4% and 0.8% HA or FA were fed for 26 days. Fe and Mn concentrations of the large intestinal content, liver, kidney, femur and hair were determined. No significant differences were observed in the production parameters. The effects of FA and HA on iron homeostasis were significantly different. FA proved to be a good iron source, and slightly increased the iron content of liver and kidney, but - up to a dietary iron level of 52.7 mg/kg - it did not influence the efficiency of iron absorption. Above a dietary iron level of 52.7 mg/kg down-regulation of Fe absorption can be assumed. HA significantly stimulated the iron uptake and there was no down-regulation of Fe absorption up to 0.8% dietary HA supplementation level (61.5 mg Fe/kg diet). In the HA groups the iron content of the liver and kidney decreased significantly, suggesting that in spite of the better Fe absorption, the HA-Fe complex does not provide iron to the investigated organs. Neither FA nor HA supplementation influenced the Fe content of the femur and hair and slightly decreased the Mn concentration in the large intestinal content. This effect was significant (with a 22.7% Mn concentration decrease) only at the HA supplementation rate of 0.8%. Neither FA nor HA influenced significantly the Mn concentrations of the liver, kidney and femur. The Mn concentration of the hair in rats receiving FA- or HA-supplemented diets was higher than in the control rats; however, this result needs further confirmation.

Effect of arginine or glutamine supplementation on production, organ weights, interferon gamma, interleukin 6 and antibody titre of broilers.

The objective of this study was designed to test whether supplementation of the diet with arginine (Arg) or glutamine (Gln) or their combination influences the production, organ weights and humoral immune response of broilers. A total of 432 one-day-old male Ross 308 broiler chickens were divided into 6 treatment groups: control, Arg-0.5%, Arg-1%, Gln-0.5%, Gln-1% and Arg-0.5%+Gln-0.5%. Drinking water and feed were provided ad libitum. On day 18 of the experiment 50% of chickens in each treatment group were immunised with bovine serum albumin. Ten and 21 days after immunisation blood samples were collected to determine the anti-albumin IgY titre, interleukin 6 (IL6) and interferon gamma (IFNG) and to measure the weight of the liver, spleen, bursa of Fabricius and thymus. Arg or Gln supplementation of the diets influenced neither the production nor the organ weights until 18 days of age. Between 18 and 39 days of age both Arg (0.5% and 1%) and Arg + Gln supplementation improved the feed conversion ratio (FCR) by 3.7%, 6.3% and 4.9%, respectively, while Gln-1% worsened it by 15%. Immunisation slightly (-0.79%) depressed the body weight gain of broilers fed the control diet, which was significantly improved by both Arg (0.5 or 1%) and Arg + Gln supplementation. Immunisation increased the weight of the spleen, bursa and thymus and decreased that of the liver. Supplementation with 1% Gln depressed (-5.13%) the body weight gain of the immunised chickens but strongly stimulated the immune response. Supplementations with Arg and Gln did not influence the IL6 and IFNG level of the blood; however, on day 10 after immunisation these two parameters showed a negative correlation with each other. Regarding production, organ weights and immunity, Arg supplementation should be recommended in the grower phase, while Gln supplementation can be useful in pullets raised for egg production, where a good immune response to vaccinations is an important factor.

Evaluation of mercury and lead content in the liver of the cormorant (Phalacrocorax carbo sinensis) population of Kis-Balaton, Hungary.

Mercury and lead concentrations were measured in the livers of cormorant (Phalacrocorax carbo sinensis), an aquatic bird species living and nesting in the special, highly protected nature conservation area of Kis-Balaton, Hungary. The measurements of metal concentrations were performed by atomic absorption spectrometry using the cold vapour method for mercury and the electrothermal method for lead. Mercury concentrations in the livers were significantly higher in the adult population (4.479 ± 3.336 mg/kg dry matter, DM) than in the juvenile birds (2.682 ± 2.087 mg/kg DM), indicating an increase of bioaccumulation with age. A similar pattern was not observed for lead. There were no statistical differences between males and females either in mercury or in lead concentrations. The average levels of mercury (3.580 ± 2.906 mg/kg DM) and lead (0.746 ± 0.499 mg/kg DM) were statistically different in the liver. No correlation was found between the concentrations of the two heavy metals. Recently, the wild birds have been chronically exposed to subtoxic amounts of metals which have a tendency to accumulate especially in the soft tissues.

Effect of dietary electrolyte balance on production, immune response and mineral concentrations of the femur in broilers.

The aim of this study was to assess the age-related effects of dietary electrolyte balance (DEB) on the performance, immune response (from day 0 to 42) and macromineral content of femur ash of broilers. The DEB values of the purchased commercial broiler diets were modified with the addition of NH4Cl or NaHCO3 to formulate the diets (DEB 325, 250, 175, 100, 25 and -50 mmol/kg) for this investigation. A total of 396 chickens were divided into 6 treatment groups and fed with the experimental diets for 6 weeks. During the first two weeks of life, DEB did not influence feed intake and body weight gain; however, by the 21st day of age DEB 175 and between 22 and 42 days of age DEB 250 mmol/kg gave significantly better results than the control. DEB did not affect the macromineral concentrations of bone ash. The immune response of broilers on low DEB (< 175 mmol/kg) was faster and more intensive than that of chickens on diets with medium or high DEB (> 175 mmol/kg). It can be concluded that the optimal DEB value required for the best body weight gain is significantly influenced by the age of broilers. Our results call attention to the discrepancy between the decreasing DEB level of commercial broiler diets and the age-related increase of 'electrolyte requirements' of broilers. It is also interesting that DEB may influence not only the performance but also the immune response of broilers.

Pathological changes induced by rancid feed in rats and effects on growth and protein utilisation.

The objective of this trial was to study the effect of rancid feeds on the health status and growth of rats and to determine the pathological changes induced by dietary rancidity. Forty-two weaned male rats (body weight: 69.3 +/- 1.0 g) were divided into seven experimental groups (n = 6 each). Rats in the different groups were fed diets containing meat-and-bone meal at an inclusion rate of 19 to 22%, low or high in peroxides and high in organic acids, with or without antioxidant. The diets were isoproteic (10%) and isolipidic (6%). During the 26-day-long trial (5 days for adjustment and 21 days for the main period) the body weight gain and the feed consumption were recorded. At the end of the trial detailed gross and histopathological examinations were performed. Feeding high-peroxide feed mixtures for 21 days significantly (P < 0.05) decreased feed intake and liveweight gain, while high organic acid concentration had only slight negative effects. Antioxidant supplementation alleviated the harmful effects, especially in the high-peroxide group. The ingestion of rancid feed mixtures and the subsequent decreased feed intake caused a decrease of glycogen content in the hepatocytes, accompanied by a slight centrolobular fatty infiltration. Peroxides caused lymphocyte depletion in the spleen, decreased the size of Malpighian bodies and the number of lymphoblasts, and altered the spermatogenesis. The protective effect of the antioxidant mixture seemed to be negligible in this respect.

Study of the energy and protein balance of pregnant rabbit does using two comparative methods.

The total body composition of 4 x 10 female, 5-month-old New Zealand White rabbits was determined, using direct chemical analysis and computerised tomography (CT). Ten non-pregnant rabbits (control) and 3 x 10 pregnant rabbits at days 14, 21 and 28 of gestation, respectively, were included in the study. With the advancement of pregnancy, the body weight (BW) of does significantly (P < 0.05) increased (control: 3255 g, day 28: 3938 g), while the dry matter, DM (control: 43.02%, day 28: 40.94%), ether extract, EE (control: 20.15%, day 28: 18.13%), crude protein, CP (control: 20.15%, day 28: 18.13%), and energy concentration decreased (control: 12.41 MJ/kg BW, day 28: 11.81 MJ/kg BW). During pregnancy the protein content of uteri (control: 12.24%, day 28: 49.45% of DM) and the retained energy in the form of uterus with fetuses (day 14: 0.824 MJ, day 28: 2.189 MJ) increased, while the dry matter (control: 82.39%, day 28: 28.96%) and ether extract content (control: 86.05%, day 28: 45.92% of DM) of uteri decreased. With the advancement of pregnancy the dry matter (day 14: 39.70%, day 28: 18.65%) and crude protein (day 14: 75.74%; day 28: 63.63% of DM) content and the number of fetuses (day 14: 8.8; day 28: 6.4) decreased, while their fat concentration (day 14: 10.13%, day 28: 24.34% of DM) increased. According to the equations created from the CT pictures, body fat (R2 = 0.871) and energy (R2 = 0.926) content can be predicted with high accuracy. The accuracy of predicting body protein content is much lower (R2 = 0.356), which can be improved by introducing the body weight as a new dependent variable (R2 = 0.797). It can also be concluded that, in contrast to other animal species, rabbit fetuses show considerable development not only in the last trimester of gestation but already from the second half of pregnancy.

Effect of inorganic and organic manganese supplementation on the performance and tissue manganese content of broiler chicks.

The effects of dietary levels of manganese (Mn) in inorganic (MnO) and organic (Mn fumarate) forms were evaluated on cockerel chicks. A basal corn-soybean diet with 23 mg/kg Mn was supplemented with levels of 0, 30, 60 and 240 ppm Mn from both Mn sources. Each treatment was replicated in five pens of 10 chicks. The chicks were fed diets ad libitum from 14 to 49 days of age, after which five birds per treatment were sacrificed for pathomorphological examinations and analysis. The treatments did not exert significant effects on the body weight (BW), the feed/gain (F/G) ratio or the mortality rate. According to the necropsy findings, no growth retardation or emaciation occurred in either of the groups and the differences in the average absolute and relative organ weights were not significant (P > 0.05). Tissue analysis indicated that the tibia showed the greatest response to Mn, followed by the liver and kidney. Accumulation in the tibia was higher (P < 0.05) with supplements of 30, 60 and 240 mg/kg from both Mn sources (3.71, 3.78, 4.44, and 3.68, 4.00, 4.36 mg/kg DM, MnO and Mn fumarate, respectively) compared to the control group (3.21 mg/kg). Accumulation in the liver increased significantly (P < 0.05) only with supplements of 60 and 240 ppm independently of the Mn source (12.7, 14.2, and 14.0, 14.9 mg/kg, respectively) compared to the control (9.8 mg/kg). Similarly, kidney tissue Mn was higher (P < 0.05) only with supplements of 60 and 240 ppm (12.8, 12.8, and 13.1, 12.5 mg/kg, respectively) compared to the control (10.2 mg/kg). At the same level of supplementation of the two Mn sources there were no significant differences (P > 0.05) between the Mn concentrations of organs and tissues. Droppings sensitively reflected the intake, whereas blood plasma and feathers showed only the extreme Mn loading.

Large doses of zinc oxide increases the activity of hydrolases in rats.

The effect of pharmacological doses of zinc oxide (1000; 2500; 5000 mg per kg diet) and two levels of dietary protein on pancreatic and intestinal hydrolase activity in rats were studied. It was hypothesized that ZnO would increase intestinal and pancreatic hydrolase enzyme activity. Male Wistar rats, averaging 64 g body weight, were randomly allocated to dietary treatments (chow diets- meeting all NRC requirements) containing 10% or 15% protein supplemented with additional ZnO (above 100 mg/kg ZnSO(4)) as follows: 0.0; 0.1; 0.25; 0.5% w/w. Water and food were provided ad libitum. Animals were fed the diets for 10 days and body weights were recorded; after decapitation blood and organ samples were collected. Amylase, lipase, trypsin, and total protease activity of pancreatic homogenates and small intestinal contents were determined. ZnO supplementation dose dependently increased the plasma Zn concentration and significantly increased amylase, lipase, trypsin and total protease activity in pancreatic homogenates and small intestinal contents. The statistical analysis showed significant protein and ZnO interaction on the activity of amylase in the pancreas, and amylase, trypsin and total-protease in the small intestinal content. Therefore ZnO at high dietary concentration may influence the digestion of nutrients via increased hydrolase activity.

Influence of feeding intensity on the growth, body composition and sexual maturity of male New Zealand White rabbits.

An experiment was carried out with young male New Zealand White (NZW) rabbits to establish live body weight changes, body measurements, body composition and sexual maturity as a function of feeding intensity. Animals in Group 1 ('AL', n = 10) were fed ad libitum, while those in Group 2 ('RS', n = 10) received restricted feeding corresponding to 70% of the ad libitum level. The starting liveweights were practically the same (0.907 +/- 0.146 and 0.911 +/- 0.147 kg in Group AL and Group RS, respectively). The feeding trial lasted from 6 to 22 weeks of age. The average body weight was significantly higher in Group AL from 7 to 22 weeks of age. At 22 weeks of age the body weight of RS rabbits was 85.64% of the weight of AL animals (3.22 +/- 0.52 kg and 3.76 +/- 0.33 kg, respectively). Average body weights of RS males at 8, 9, 11, 19 and 21 weeks of age were similar to those of ad libitum fed (AL) animals at 7, 8, 10, 15 and 16 weeks of age, respectively. The growth of bucks fed restricted tended to be allometric. The most significant difference was found at 16 and 18 weeks of age, while the lowest difference occurred at 6, 12, 15 and 19 weeks of age. It can be stated that low-intensity feeding up to slaughtering weight causes backwardness in rear cannon length and this backwardness remains also after the 15th week, which is well over the optimal slaughtering age. Based on the present data, the 70% restricted feeding cannot be recommended either for the future breeding bucks or for broiler males reared for slaughter. To determine the major chemical components of the body, rabbits were euthanised. Original dry matter and crude fat content of the body significantly (P < 0.05) decreased under restricted feeding (41.42%; 32.48% and 16.73%; 7.35%) while the percentage of protein within the dry matter increased (49.6%; 65.0%) and fat decreased (40.17%; 22.1%) significantly. Libido unambiguously decreases as a consequence of feed deprivation. The most conspicuous difference was found in the level of blood testosterone. Although a few RS bucks produced semen but only much later than the rabbits fed ad libitum. On the other hand, there was no difference in the motility of spermatozoa and ejaculate volume in comparison with AL animals. There was no relationship between the body fat content and the reproductive status of bucks in the present trial.

Effect of different fat sources on in vitro degradation of nutrients and certain blood parameters in sheep.

This study was designed to determine the effects of calcium salt of palm oil fatty acids (CS), hydroxyethylsoyamide (HESA), butylsoyamide (BSA) and soybean oil (SO) on degradation of crude protein and fibre in vitro, and on the blood plasma lipid parameters in vivo. Five mature wethers (body weight 75 kg) were fed five diets in a 5 x 5 Latin square experiment. The control diet consisted of 50% meadow hay and 50% concentrate with no added fat. The control diet was supplemented with CS, HESA, BSA, or SO. Fat was added at 3.5% of dietary dry matter (DM). The final ether extract content of the ration was near 6%. Each period lasted 20 days. Fat supplements, except HESA, consistently decreased the in vitro DM disappearance of soybean meal as compared to control. In contrast to the effect of other treatments, crude protein degradation was greatest in the test tubes with inocula obtained from sheep fed diet with HESA. Fat supplements equally inhibited the DM and fibre breakdown of alfalfa pellet. CS and HESA seemed to be less detrimental to in vitro fermentation of neutral detergent fibre (NDF) than BSA and SO. All fat supplements increased blood plasma triglyceride, cholesterol and total lipid content. Plasma concentration of cholesterol and total lipid was highest with SO. The inclusion of CS in the diet increased 16:0, while all fat supplements increased plasma 18:0 and decreased 16:1 and 18:1 fatty acid content. Plasma 18:2n-6 was not changed by feeding CS and SO. However, compared to the control diet, 18:2n-6 increased with 12 and 41% in plasma fatty acids when sheep were fed HESA and BSA, respectively. The results showed that plasma concentration of linoleic acid was enhanced more when the amide was synthesised from butylamine than when from ethanolamine.