Comparison between Organic and Inorganic Zinc Forms and Their Combinations with Various Dietary Fibers in Respect of the Effects on Electrolyte Concentrations and Mucosa in the Large Intestine of Pigs.

This study aimed to determine the effects of Zn sources, used with potato fiber (PF) or lignocellulose (LC), on electrolyte concentration and the mucus layer in the large intestine of pigs. The experiment involved 24 barrows with an initial body weight of 10.8 ± 0.82 kg, divided into four groups fed the following diets: LC and ZnSO4, LC and Zn glycinate (ZnGly), PF and ZnSO4, or PF and ZnGly. Fiber supplements provided 10 g crude fiber/kg diet, while Zn additives introduced 120 mg Zn/kg diet. After four weeks of feeding, the pigs were sacrificed and digesta and tissue samples were taken from the cecum and colon. PF increased the water content and decreased the phosphorus concentration in the large intestine in comparison with LC. PF also increased calcium, iron, and chloride concentrations in the descending colon. Mucus layer thickness and histological parameters of the large intestine were not affected. ZnGly diets increased MUC12 expression in the cecum as compared to the LC-ZnSO4 group. In the ascending colon, the PF-ZnGly diet increased MUC5AC expression, while both PF groups had greater MUC20 expression in comparison with the LC-ZnSO4 group. In the transverse colon, the LC-ZnGly group and both PF groups had higher MUC5AC expression in comparison with the LC-ZnSO4 group, and both ZnGly groups had higher MUC20 expression than ZnSO4 groups. PF and ZnGly increased MUC4 and MUC5AC expression in the descending colon. PF and ZnGly may exert a beneficial effect on colon health in pigs by upregulating the expression of the MUC5AC and MUC20 genes and are more effective than LC and ZnSO4.

Impact of Sainfoin (Onobrychis viciifolia) Pellets on Parasitological Status, Antibody Responses, and Antioxidant Parameters in Lambs Infected with Haemonchus contortus.

Our study analyzed the parasitological status, antibody responses, and antioxidant parameters of lambs experimentally infected with a gastrointestinal nematode during the consumption of sainfoin pellets (SFPs) for 14 d. Twenty-four lambs infected with Haemonchus contortus were separated into two groups: untreated animals (control) and animals treated with SFPs (600 g dry matter/d). SFP treatment began on day (D) 30 post-infection. The number of eggs per gram (EPG) of feces was quantified on D18, D23, D26, D30, D33, D37, D40, and D44. The mean reductions in EPG on D40 and D44 were 33.6 and 36.7%, respectively. The number of abomasal worms was lower for the SFP than the control group (p < 0.05). SFP treatment did not significantly affect either the total or the local antibody response (p > 0.05). The blood activity of glutathione peroxidase was affected by the treatment (p < 0.022). Adult worms were selected for scanning electron microscopy after necropsy, but surface structures of adult H. contortus females did not differ between the groups. The treatment of lambs with SFPs directly affected the dynamics of infection, probably indirectly by mobilizing the antioxidant defensive system and antibody response thus improving animal resistance.

Influence of the Zinc and Fibre Addition in the Diet on Biomechanical Bone Properties in Weaned Piglets.

The effects of the zinc and fibre source in piglets' diet on the bone mineral content, density, and strength parameters of the femur were investigated using 24 piglets fed a diet supplemented with either lignocellulose (LC) or potato fibre (PF). Half of each group of piglets consumed a diet with ZnSO4 monohydrate or with zinc glycinate (ZnGly). The diets contained similar amounts of lysine, energy, and fibre. Bone mineral content and density were over 9% higher in pigs receiving diets with ZnGly than in animals fed diets with ZnSO4. Moreover, ZnGly strongly improved maximum and elastic strength (by 25.7 and 20.0%, respectively, p < 0.0001) and bone stiffness (by 29.4%, p < 0.0001). Only the mass of the femur was affected by the type of fibre in the diet, as the femurs of piglets fed diets with LC were over 7% (p < 0.0001) heavier than in piglets fed diets with PF. The intake of digestible zinc and the zinc content in the blood serum were positively correlated with the measured bone parameters and, depending on the parameter, "r" ranged from 0.749 to 0.866 and from 0.400 to 0.479, respectively. It can be concluded that bone parameters are affected more strongly by the organic than inorganic source of zinc.

Changes in the Antioxidant and Mineral Status of Rabbits After Administration of Dietary Zinc and/or Thyme Extract.

This study was aimed at determining the impact of organic zinc (Zn) and thyme extract (TE) administration, given alone or together for 6 weeks, on the antioxidant and mineral status (Zn, Cu, Fe, and Mn) in the plasma and tissues of growing rabbits. A total of 96 rabbits of age 35 days were randomly assigned to one of four treatment groups: a control group (C), a Zn group supplemented with dietary zinc (50 mg/kg), a TE group receiving thyme extract applied in drinking water (1 ml/L), and a Zn + TE group treated with both additives. Lipid peroxidation in the plasma was influenced by Zn intake and in the kidney was affected by both the Zn and TE treatment (P < 0.05). Zn supplementation led to a significant increase in glutathione peroxidase activity (P = 0.017), total antioxidant capacity (P = 0.009) and total thiol groups level (P = 0.047) in the kidney, with the highest values occurring in rabbits receiving the combination Zn + TE. Administration of TE influenced Zn content in the kidney (P < 0.001), while zinc intake decreased Cu concentration in muscle (P = 0.021). In conclusion, the simultaneous administration of organic Zn and TE positively affected the antioxidant response of kidneys and can be used for improving the antioxidant status of growing rabbits.

Effects of Medicinal Plants and Organic Selenium against Ovine Haemonchosis.

The objective of this study was to determine the effect of traditional medicinal plants typical to Central Europe as well as organic selenium on increasing the resistance of lambs to gastrointestinal nematode (GIN) infection with Haemonchus contortus. 21 female lambs were infected with third-stage larvae of H. contortus on the day (D) 0 and re-infected on D49 and D77. The animals were divided into three groups based on a treatment diet: a basal diet (control), a diet enriched with dry plants (Herbmix), and a diet enriched with selenized yeast (Selplex). The number of eggs per gram (EPG) of feces was quantified on D21, D28, D35, D42, D49, D56, D63, D70, D77, D84, D91, D98, D105, D112, and D119. The mean reductions in EPG on D28 were 43.4 and 28.6% for Selplex and Herbmix, respectively. The reduction in egg output was nearly uniform throughout the experiment for Selplex. However, for Herbmix the mean reduction was only 19.8% up to D91 and 46.1% after D91. Glutathione peroxidase activity in the blood from D35 to D98 was two to three-fold higher for Selplex than the other groups. Both supplements slowed the dynamics of GIN infection and gradually increased the resistance of lambs against ovine haemonchosis.

Effect of Hydrolyzed Yeast Administration on Faecal Microbiota, Haematology, Serum Biochemistry and Cellular Immunity in Healthy Dogs.

Fungal microorganisms are regularly found in the gastrointestinal tract of healthy and diseased dogs especially from the phyla Ascomycota and Basidiomycota; however, it is necessary to better understand their role in host health. One of the most commonly studied yeast species in humans or animals is Saccharomyces cerevisiae especially in its live cell form. Scarce knowledge on its hydrolysate product effects in dogs forced us to test diet supplemented with hydrolyzed brewery S. cerevisiae (at a dose 0.3% of the diet) for 14 days to healthy adult dogs. Twenty German Shepherds were randomly divided into 2 groups: control and experimental, ten dogs in each. The experiment lasted 42 days (blood and faeces sample collection at days 0, 14, 28 and 42). The results of this straighforward experiment showed significant increase in the abundance of bifidobacteria (day 14), lactic acid bacteria (day 42) and clostridia (day 42). The faecal pH was significantly increased at day 28. In blood serum, the concentration of triglyceride and cholesterol decreased (day 42) while activities of alanine aminotransferase (at day 14) and aspartate aminotransferase significantly increased (at days 28 and 42). Activities of these enzymes were above reference range top in 7 dogs (ALT) and 4 dogs (AST). Haematological paramaters and activity of phagocytes as well as on percentage of lymphocyte subsets CD4+, CD8+, CD4+CD8+ and CD21+ were not changed during the experiment. The important point of these results is their onset mostly in the post-supplementation period. The observation of some unexpected effects emphasizes the need for reassessment to use yeasts products for dogs but further studies using different doses are necessary.

Corrigendum: Impact of Zinc and/or Herbal Mixture on Ruminal Fermentation, Microbiota, and Histopathology in Lambs.

[This corrects the article DOI: 10.3389/fvets.2021.630971.].

Impact of Zinc and/or Herbal Mixture on Ruminal Fermentation, Microbiota, and Histopathology in Lambs.

We investigated the effect of diets containing organic zinc and a mixture of medicinal herbs on ruminal microbial fermentation and histopathology in lambs. Twenty-eight lambs were divided into four groups: unsupplemented animals (Control), animals supplemented with organic zinc (Zn, 70 mg Zn/kg diet), animals supplemented with a mixture of dry medicinal herbs (Herbs, 100 g dry matter (DM)/d) and animals supplemented with both zinc and herbs (Zn+Herbs). Each lamb was fed a basal diet composed of meadow hay (700 g DM/d) and barley (300 g DM/d). The herbs Fumaria officinalis L. (FO), Malva sylvestris L. (MS), Artemisia absinthium L. (AA) and Matricaria chamomilla L. (MC) were mixed in equal proportions. The lambs were slaughtered after 70 d. The ruminal contents were used to determine the parameters of fermentation in vitro and in vivo and to quantify the microbes by molecular and microscopic methods. Samples of fresh ruminal tissue were used for histopathological evaluation. Quantitative analyses of the bioactive compounds in FO, MS, AA, and MC identified 3.961, 0.654, 6.482, and 12.084 g/kg DM phenolic acids and 12.211, 6.479, 0.349, and 2.442 g/kg DM flavonoids, respectively. The alkaloid content in FO was 6.015 g/kg DM. The diets affected the levels of total gas, methane and n-butyrate in vitro (P < 0.046, < 0.001, and < 0.001, respectively). Relative quantification by real-time PCR indicated a lower total ruminal bacterial population in the lambs in the Zn and Zn+Herbs groups than the Control group (P < 0.05). The relative abundances of Ruminococcus albus, R. flavefaciens, Streptococcus bovis, and Butyrivibrio proteoclasticus shifted in the Zn group. Morphological observation found a focally mixed infiltration of inflammatory cells in the lamina propria of the rumen in the Zn+Herbs group. The effect of the organic zinc and the herbal mixture on the parameters of ruminal fermentation in vitro was not confirmed in vivo, perhaps because the ruminal microbiota of the lambs adapted to the zinc-supplemented diets. Long-term supplementation of a diet combining zinc and medicinal herbs, however, may negatively affect the health of the ruminal epithelium of lambs.

Effect of Artemisia absinthium and Malva sylvestris on Antioxidant Parameters and Abomasal Histopathology in Lambs Experimentally Infected with Haemonchus contortus.

This study evaluated the effect of Artemisia absinthium and Malva sylvestris on antioxidant response and histopathological changes in the abomasa of the Haemonchus contortus infected lambs. Twenty-four lambs were divided into four groups: unsupplemented lambs (UNS), lambs supplemented with A. absinthium (ART), lambs supplemented with M. sylvestris (MAL), and lambs supplemented with both plants (ARTMAL). Lambs were infected orally with approximately 5000 third-stage (L3) larvae of H. contortus. The experiment was conducted for 75 d (days), all animals were then slaughtered; and the abomasal tissues were examined for antioxidant parameters and histopathology. The concentration of malondialdehyde in the abomasal mucosa was lower in ARTMAL (p < 0.05), and the total antioxidant capacity was higher in MAL (p < 0.05), than in UNS. Increased mucus production was observed in the ARTMAL. The number of mast cells in UNS and ART was significantly higher than the number in MAL (p < 0.01 and p < 0.05). Plasma cell numbers were higher in ARTMAL than the number in MAL (p < 0.05). Abomasal tissue regenerated more frequently in ARTMAL. These results represent the first report of the impact of A. absinthium and M. sylvestris on antioxidant parameters and local immune responses of abomasal mucosa of lambs infected with a GIN parasite.

Does Herbal and/or Zinc Dietary Supplementation Improve the Antioxidant and Mineral Status of Lambs with Parasite Infection?

This study was conducted to evaluate the effect of feed supplementation with a medicinal herbs mixture (Hmix) and organic zinc (Zn), alone or in combination, on the antioxidant responses and mineral status of lambs infected with the gastrointestinal nematode parasite Haemonchus contortus. A total of 24 experimentally infected lambs were randomly allocated to 1 of 4 dietary treatments (n = 6). The diets included an unsupplemented control diet (CON) and the CON further supplemented with Hmix, Zn, or both Hmix + Zn. Antioxidant enzymes activities, lipid peroxidation, total antioxidant capacity (TAC) and microelement (Zn, Cu, Fe, Mn) concentrations were analyzed in serum, liver, kidney, and intestinal mucosa. Zinc treatment elevated the superoxide dismutase activities in the duodenal mucosa and ileal TAC. Intake of Hmix resulted in higher kidney and ileal catalase activity and also influenced the TAC of the liver and intestinal mucosa. The inclusion of Hmix or Zn alone into the diet increased glutathione peroxidase activity in the blood, liver and duodenal mucosa. Tissue mineral uptake was not affected by herbal supplementation. Organic Zn intake increased the serum and liver Zn levels and influenced the Cu concentration in duodenal mucosa. Dietary supplementation with Hmix and/or Zn might promote the antioxidant status of lambs infected with Haemonchus spp.

Ruminal fermentation, microbial population and lipid metabolism in gastrointestinal nematode-infected lambs fed a diet supplemented with herbal mixtures.

The aim of this study was to evaluate the effects of medicinal herbal mixtures rich in phenolic, flavonoid and alkaloid compounds on ruminal fermentation and microbial populations, and fatty acid (FA) concentrations and lipid oxidation in tissues of lambs infected with the gastrointestinal nematode (GIN) parasite (Haemonchus contortus). Parallel in vitro and in vivo studies were performed using two different herbal mixtures (Mix1 and Mix2). The in vitro study was conducted in a 2 (infection status; non-infected versus infected) × 3 (diets; control, Mix1 and Mix2) factorial design. In the in vivo study, 24 lambs were equally divided into four treatments: non-infected lambs fed a control diet, infected lambs fed the control diet, infected lambs fed a diet with Mix1 and infected lambs fed a diet with Mix2. Herbal mixtures (100 g dry matter (DM)/d) were added to the basal diets of meadow hay (ad libitum) and a commercial concentrate (500 g DM/d). The experimental period lasted for 70 days. Ruminal fermentation characteristics and methane production were not affected by infection in vivo or in vitro. Both herbal mixture supplementation increased total volatile fatty acid (VFA) concentrations (P < 0.01) and DM digestibility (P < 0.01) in vitro. Archaea population was slightly diminished by both herbal mixtures (P < 0.05), but they did not lower methane production in vitro or in vivo (P > 0.05). Infection of H. contortus or herbal mixtures modulated FA proportion mainly in the liver, especially the long chain FA proportion. Concentrations of thiobarbituric acid reactive substances (TBARS) in serum were significantly higher after 70 days post-infection in the infected lambs. Herbal Mix1 supplementation reduced TBARS concentrations in meat after seven days of storage. In conclusion, supplementing of herbal mixtures to the diets of GIN parasite infected lambs did not affect the basic ruminal fermentation parameters. Herbal mixtures may improve few FA proportions mainly in liver as well as decrease lipid oxidation in meat.

Effect of dry medicinal plants (wormwood, chamomile, fumitory and mallow) on in vitro ruminal antioxidant capacity and fermentation patterns of sheep.

The objective of this study was to determine the effect of dry medicinal plants (wormwood, chamomile, fumitory and mallow) and dietary substrates containing a mix of the plants on the end products of in vitro ruminal and intestinal fermentation, rumen protozoan population and ruminal antioxidant capacity of sheep. The experiment consisted of fermentations with the four plants used individually as the sole substrate and fermentation of a mix of medicinal plants (Plants): meadow hay:barley grain (MH:B), 700/300 w/w and Plants:MH:B, 100/600/300 w/w/w. The experiment was conducted using the in vitro gas production technique (IVGPT) with 35 ml of buffered inocula and approximately 250 mg (DM basis) of substrate incubated for 24 hr at 39°C in anaerobic conditions. Quantitative analyses of the bioactive compounds by ultra-high-resolution mass spectrometry in Plants identified three main groups: flavonoids (22 mg/g DM), phenolic acids (15 mg/g DM) and alkaloids (3 mg/g DM). The total antioxidant capacity (TAC) of the plant extracts and rumen fluid was analysed using a ferric reducing antioxidant power assay. The values of total and individual short-chain fatty acids, acetate:propionate ratio, pH and total gas production were significantly affected by the single plant substrates and inocula (p < .001). Apart from these parameters, the values of ammonia N, methane production and total gas produced were decreased in Plants:MH:B in comparison with MH:B (p < .001). A positive correlation was recorded between total polyphenols content and TAC of plant extracts (R2  = .778, p < .001). The counts of the total ciliate protozoan population in rumen fluid after 24 hr of fermentation were not significantly different (p > .05). Results suggest that the dietary substrate containing the medicinal plant mix possessed strong ruminal antioxidant capacity, had the potential to reduce methane emission and ammonia concentration and caused desirable changes in the gastrointestinal ecosystem.

Anthelmintic Activity of Wormwood (Artemisia absinthium L.) and Mallow (Malva sylvestris L.) against Haemonchus contortus in Sheep.

The objective of this study is to evaluate the effect of dry wormwood and mallow on the gastrointestinal parasite of small ruminants Haemonchus contortus. Twenty-four experimentally infected lambs were randomly divided into four groups of six animals each: unsupplemented lambs, lambs supplemented with wormwood, lambs supplemented with mallow and animals supplemented with a mix of both plants. Faecal samples from the lambs were collected on day 23, 29, 36, 43, 50, 57, 64 and 75 post-infection for quantification of the number of eggs per gram (EPG). The mix of both plants contained phenolic acids (10.7 g/kg DM) and flavonoids (5.51 g/kg DM). The nematode eggs were collected and in vitro egg hatch test was performed. The aqueous extracts of both plants exhibited strong ovicidal effect on H. contortus, with ED50 and ED99 values of 1.40 and 3.76 mg/mL and 2.17 and 5.89 mg/mL, respectively, in the in vitro tests. Despite the great individual differences between the treated lambs in eggs reduction, the mean EPG of the untreated and treated groups did not differ (p > 0.05). Our results indicate that using wormwood and mallow as dietary supplements do not have a sufficient effect on lambs infected with H. contortus.

Natural chemotherapeutic alternatives for controlling of haemonchosis in sheep.

BACKGROUND: Parallel in vitro and in vivo experiments were designed to evaluate promising chemotherapeutic alternatives for controlling haemonchosis in ruminants. In vitro anthelmintic activities (egg hatch test - EHT; larval development test - LDT) of aqueous and methanolic herbal extracts Mix1 and Mix2 were investigated. The in vivo effects of dietary supplementation with Mix1 and Mix2 on the parasitological status, inflammatory response, antioxidant parameters and microbial community of the lambs infected experimentally with Haemonchus contortus were investigated. Lambs were divided into four groups for the in vivo study: uninfected control lambs (C), infected lambs (I), infected lambs supplemented with Mix1 (I + Mix1) and infected lambs supplemented with Mix2 (I + Mix2). The experimental period was 70 days. RESULTS: The number of eggs per gram (EPG) of feces was quantified 22, 30, 37, 44, 51, 58, 65 and 70 days post-infection, and mean abomasal worm counts were assessed 70 days post-infection. Quantitative analyses identified 57.3 and 22.2 mg/g phenolic acids, 41.5 and 29.5 mg/g flavonoids and 1.4 and 1.33 mg/g protoberberine-type alkaloids in Mix1 and Mix2, respectively. The methanolic extracts of the herbal mixtures in both in vitro tests had higher anthelmintic effects (P < 0.01) than the aqueous extracts, but the effects did not differ significantly between Mix1 and Mix2 (P > 0.05). I + Mix1 and I + Mix2 lowered mean EPGs between 44 and 70 d by 58.1 and 51.6%, respectively. The level of IgG antibodies against H. contortus increased significantly after infection in each infected group. CONCLUSION: These results represent the first monitoring of the in vitro anthelmintic effects of herbal mixtures on H. contortus. The in vivo experiment indicated that the anthelmintic effect was not sufficient for the elimination of parasites, but this herbal treatment may affect the host over a longer term, reducing the parasitic infection in the host.

Dietary Zinc and Fibre Source can Influence the Mineral and Antioxidant Status of Piglets.

The study investigated the effect of dietary zinc glycine chelate and potato fibre on the absorption and utilisation of Zn, Cu, Fe, and Mn; the activity of Zn-containing enzymes (superoxide dismutase, SOD; alkaline phosphatase, ALP); and zinc transporter concentrations (metalothionein1, MT1; zinc transporter1, ZnT1) in tissues, with a special emphasis on the small intestine. Twenty-four barrows (Danbred × Duroc) were randomly allotted to four diets (supplemented with 10 g/kg of crude fibre and 120 mg Zn/kg) that consisted of cellulose and either zinc sulphate (C) or zinc glycinate (ZnGly), or contained potato fibre supplemented with ZnSO4 (PF) or ZnGly (PF + ZnGly). Feeding PF can influence the Zn absorption in the small intestine due to reduced zinc transporters MT1 and ZnT1 in the jejunum. The activity of antioxidant enzyme SOD and liver ZnT1, and duodenal iron concentrations were increased in the PF treatments. Dietary ZnGly did not significantly influence the Zn distribution, but it may alter the absorption of Fe and Mn. Given the elevated content of thiol groups and the Zn/Cu ratio in plasma, as well as the altered SOD activity and MT content in the tissues, we can conclude that feeding PF and ZnGly can influence the mineral and antioxidant status of growing piglets. However, further research is needed in order to elucidate the effect of both dietary sources on the transport systems of other minerals in enterocytes.

Modulation of ruminal and intestinal fermentation by medicinal plants and zinc from different sources.

Two experiments were conducted on sheep to determine the effect of dietary supplementation with zinc and a medicinal plant mixture on haematological parameters and microbial activity in the rumen and large intestine. In Experiment 1, 24 male lambs were randomly divided into four groups: One group was fed an unsupplemented basal diet (control), and three groups were fed a diet supplemented with 70 mg Zn/kg diet in the form of Zn sulphate (ZnSO4 ), a Zn-chelate of glycine hydrate (Zn-Gly) or a Zn-proteinate (Zn-Pro), for five months. The ruminal content was collected separately from each lamb, and batch cultures of ruminal fluid were incubated in vitro with mixture of medicinal plants (Mix) with different roughage:concentrate ratios (800:200 and 400:600, w/w). Bioactive compounds in Mix were quantified by UPLC/MS/MS. In Experiment 2, four sheep were fed a diet consisting of meadow hay and barley grain (400:600, w/w), with Zn-Gly (70 mg Zn/kg diet), Mix (10% replacement of meadow hay) or Zn-Gly and Mix (Zn-Gly-Mix) as supplements in a Latin square design. Mix decreased total gas (p < 0.001) and methane (p < 0.01) production in vitro. In Experiment 1, caecal isobutyrate and isovalerate concentrations varied among the dietary treatments (p < 0.01). The isovalerate concentration of the zinc-supplemented groups in the distal colon was higher (p < 0.001) compared with the control. In Experiment 2, the molar proportion of isobutyrate was the highest in the faeces of the sheep fed the diet with Zn-Gly-Mix (p < 0.01). The plasma zinc concentration was higher in the groups fed a diet supplemented with zinc (p < 0.001). The haematological profile and antioxidant status did not differ between the dietary groups (p > 0.05). The diets containing medicinal plants and organic zinc thus helped to modulate the characteristics of fermentation in ruminants.

Effects of herbal nutraceuticals and/or zinc against Haemonchus contortus in lambs experimentally infected.

BACKGROUND: The gastrointestinal parasitic nematode Haemonchus contortus is a pathogenic organism resistant to several anthelmintics. This study assessed the efficacy of a medicinal herbal mixture (Herbmix) and organic zinc, as an essential trace element for the proper functioning of both unspecific and specific immune defensive mechanisms, against experimental infections with H. contortus in lambs. All lambs were infected orally with approximately 5000 third-stage larvae of a strain of H. contortus susceptible to anthelmintics (MHco1). Twenty-four female lambs 3-4 months of age were divided into four groups: unsupplemented animals (control), animals supplemented with Herbmix (Hmix), animals supplemented with organic zinc (Zn) and animals supplemented with Herbmix and organic zinc (Hmix+Zn). Eggs per gram (EPG) of faeces were quantified 20, 28, 35, 42, 49, 56, 62 and 70 d post-infection and mean abomasal worm counts were assessed 70 d post-infection. Samples of blood were collected from each animal 7, 35, 49 and 70 d post-infection. RESULTS: Quantitative analyses of the bioactive compounds in Herbmix identified three main groups: flavonoids (9964.7 µg/g), diterpenes (4886.1 µg/g) and phenolic acids (3549.2 µg/g). Egg counts in the lambs treated with Hmix, Zn and Hmix+Zn decreased after 49 d. The EPGs in the Zn and Hmix+Zn groups were significantly lower on day 56 (P < 0.05 and P < 0.01, respectively), and the EPGs and mean worm counts were significantly lower on day 70 in all supplemented groups (P < 0.05 and P < 0.01). Hemograms of complete red blood cells of each animal identified clinical signs of haemonchosis after day 35. Serum calprotectin concentrations and IgA levels were significantly affected by treatment. The treatment influenced serum malondialdehyde concentrations (P < 0.05) and sulfhydryl groups (P < 0.01) of antioxidant status. The mineral status was unaltered in all lambs. CONCLUSION: A direct anthelmintic impact on the viability of nematodes was not fully demonstrated, but the treatments with herbal nutraceuticals and zinc likely indirectly contributed to the increase in the resistance of the lambs to nematode infection.

The effects of high dose of two manganese supplements (organic and inorganic) on the rumen microbial ecosystem.

Little is known about the effects of the high dose and types of manganese supplements on rumen environment at manganese intake level close above the limit of 150 mg/kg of dry feed matter. The effects of high dose of two manganese supplements (organic and inorganic) on rumen microbial ecosystem after four months of treatment of 18 lambs divided into three treatment groups were studied. We examined the enzyme activities (α-amylase, xylanase, and carboxymethyl cellulase), total and differential microscopic counts of rumen ciliates, total microscopic counts of bacteria, and fingerprinting pattern of the eubacterial and ciliates population analyzed by PCR-DGGE. Lambs were fed a basal diet with a basal Mn content (34.3 mg/kg dry matter; control) and supplemented either with inorganic manganous sulfate or organic Mn-chelate hydrate (daily 182.7, 184 mg/kg dry matter of feed, respectively). Basal diet, offered twice daily, consisted of ground barley and hay (268 and 732 g/kg dry matter per animal and day). The rumens of the lambs harbored ciliates of the genera of Entodinium, Epidinium, Diplodinium, Eudiplodinium, Dasytricha, and Isotricha. No significant differences between treatment groups were observed in the total ciliate number, the number of ciliates at the genus level, as well as the total number of bacteria. Organic Mn did decrease the species richness and diversity of the eubacterial population examined by PCR-DGGE. No effects of type of Mn supplement on the enzyme activities were observed. In comparison to the control, α-amylase specific activities were decreased and carboxymethyl-cellulase specific activities were increased by the Mn supplements. Xylanase activities were not influenced. In conclusion, our results suggested that the intake of tested inorganic and organic manganese supplements in excess may affect the specific groups of eubacteria. More studies on intake of Mn supplements at a level close to the limit can reveal if the changes in microbial population impact remarkably the other rumen enzymatic activities.

Effects of Different Dietary Selenium Sources on Antioxidant Status and Blood Phagocytic Activity in Sheep.

The objective of this experiment was to investigate the effects of feed supplementation with equivalent doses of selenium from sodium selenite (SS) or selenized yeast (SY) on Se deposition, selenoenzyme activity and lipid peroxidation in tissues as well as in bacterial and protozoal fractions of rumen contents in sheep. The phagocytic activity of monocytes and neutrophils in whole blood was also assessed after 3 months of dietary treatment. While animals in the control group were fed with unsupplemented basal diet (BD) containing only background Se (0.16 mg/kg DM), the diet of the other two groups (n = 6) consisted of identical BD enriched with 0.4 mg Se/kg DM either from SS or SY. Concentrations of Se in blood and tissues were found to be significantly increased in both supplemented groups. No response in Se deposition was recorded in the musculus longissimus dorsi of sheep given dietary SS. The intake of SY resulted in a significantly higher Se level in the blood, kidney medulla, skeletal muscles, heart, intestinal and ruminal mucosa than in the case of SS supplementation. No differences appeared between tissue Se contents in the liver and kidney cortex due to the source of added Se. Regardless of source, Se supplementation to feeds significantly increased the glutathione peroxidase (GPx) activity in blood and tissues except the kidney medulla and jejunal mucosa. Supplementation with SY resulted in significantly higher activity of thioredoxin reductase in the liver and ileal mucosa, and also reduced malondialdehyde content in the liver and duodenal mucosa. Dietary Se intake increased Se concentrations in the total rumen contents and bacterial and protozoal fractions. The accumulation of Se in rumen microbiota was associated with increased GPx activity. Phagocytic cell activity was enhanced by Se supplementation. Our results indicate that Se from both sources has beneficial effects on antioxidant status in sheep and can be utilized by rumen microflora.

Dietary manganese source does not affect Mn, Zn and Cu tissue deposition and the activity of manganese-containing enzymes in lambs.

Manganese (Mn) is a trace element required for normal physiological processes in animals and humans. Organic forms of trace elements are expected to have higher bioavailability in comparison with inorganic sources. The effect of feeding a diet supplemented with different sources of manganese to lambs was studied in a 112-d feeding trial. The aim of this study was to investigate the deposition of Mn in relation to activities of superoxide dismutase (SOD), manganese superoxide dismutase (MnSOD) and arginase in the tissues of lambs fed the diet supplemented with an inorganic or an organic source of manganese up to the maximum total Mn content allowed in the European Union (150mg Mn/kg). A total of eighteen female lambs of the improved Valachian breed were randomly allocated to three dietary treatments and fed an unsupplemented basal diet (Control, 31mg Mn/kg) or the identical diet supplemented with manganese sulphate (MnSO4) or manganese chelate of glycine hydrate (Mn-Gly) with a total Mn content up to 150mg/kg. Regardless of the source, feed supplementation with manganese increased Mn concentrations in plasma (P˂0.05) and the liver (P˂0.001) as well as the activity of liver MnSOD (P˂0.05) and arginase (P˂0.001) compared with the control lambs. In the kidney cortex, the concentration of Mn was greatest in lambs fed the diet supplemented with the chelated Mn source compared with animals receiving the inorganic Mn source (P˂0.05) and the unsupplemented diet (P˂0.001). The 112-d intake of feed enriched with manganese did not result in any change in Mn levels, SOD or MnSOD activity in pancreas and kidney tissues. Plasma Cu concentration was depressed in both supplemented treatments. No analyzed tissue showed a change in zinc and copper levels, except the greater Cu concentration in the liver of lambs fed the diet with Mn-Gly. The presented results did not indicate any differences between dietary Mn sources either in Mn tissue deposition or activity of SOD, MnSOD and arginase in lamb tissues; however, it seems that manganese intake of up to 150mg/kg in feed may decrease plasma Cu concentration in lambs.