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The starch content of triticale and oat grains provides much of their readily available energy. Synchronizing energy and nitrogen in the rumen is important in optimizing profitability; for this reason, ammonia processing of these grains was evaluated for its potential to modify ruminal fermentation and to improve milk production performance. A mixture of ground triticale and oats (CONG, at a ratio of 60:40 on a dry matter basis) was treated with urea (5 kg/1000 kg) and urease additive (20 kg/1000 kg) containing 200 g/kg of moisture, for 2 wk (UREG). The urea treatment enhanced the pH and CP content of grains by 34% and 52%, respectively. In a batch culture study, CONG or UREG as the only substrate was incubated in a buffered ruminal fluid. Compared to CONG, UREG increased pH, total VFA concentration, total gas, and disappearance of DM, while reducing CH4 production, whereas NH3 concentration increased and entodiniomorph counts tended to increase. In the in vivo study, cows were randomly allocated to two dietary groups (n = 24) and were offered TMR based on maize and grass silage, containing either 155 g/kg of CONG and 80 g/ kg of soybean meal (CONT) or 155 g/kg of UREG and 59 g/kg of soybean meal (URET) for 31 d. Ruminal fluid was collected (n = 10) using rumenocentesis. The relative abundances of Streptococcus bovis decreased, but Megasphaera elsdenii, methanogens, and ammonia-producing bacteria increased by URET. Entodiniomorph and holotrich counts were decreased by URET. Feeding with URET increased ruminal pH and concentrations of total VFA, acetate, branched-chain VFA, and NH3. Feeding with URET also increased milk yield. These results demonstrate that replacing untreated triticale and oat grains with urea-treated grains can beneficially modulate ruminal microbiota and fermentation, consequently improving production performance and profitability.

Article Details: Received: 2020-10-06 | Accepted: 2020-11-27 | Available online: 2021-01-31 https://doi.org/10.15414/afz.2021.24.mi-prap.29-32 The aim of this experiment was to determine the effect of medicinal plants ( Artemisia absinthium and Malva sylvestris ) as feed supplements on blood parameters, and local immune responses in the abomasum of Haemonchus contortus infected lambs. Twenty-four lambs were infected orally with 5000 L3 larvae of H. contortus and subsequently randomly divided into four groups: unsupplemented animals (UNS), animals with A. absinthium (ART), animals with M. sylvestris (MAL), and animals with a mix of two plants (ARTMAL). During the experiment, samples of blood were taken for evaluation of hemoglobin and albumin levels. All animals were humanely killed after 75 days of the experiment, and abomasum tissue samples were taken for histopathology. The hemoglobin concentration in the blood was influenced by time ( p< 0.001) and in all groups decreased until day 45. The serum albumin levels were influenced by treatment ( p< 0.05) and time ( p< 0.001) and in all groups occurred reduction until the end of the experiment compared to day 15. The number of plasma cells was higher in ARTMAL compared to MAL ( p < 0.05). UNS and ART had significantly higher numbers of mast cells compared to MAL ( p< 0.01 and 0.05, respectively). The regeneration of abomasal tissue was most frequent in ARTMAL. The results showed that dietary supplementation by A. absinthium and M. sylvestris did not positively affected blood parameters but influenced local immune response in the abomasum of H. contortus infected sheep. Keywords: Haemonchus contortus , sheep, hemoglobin, abomasum, local immune response References Besier, R. et al. (2016). The Pathophysiology, Ecology and Epidemiology of Haemonchus contortus. In Gasser, RB and Samson-Himmelstjerna, G. Haemonchus contortus- Past, present and future trends. (1. ed.). London: Elsevier Ltd (pp. 95-143). Hoste, H. et al. (2016). Interactions between nutrition and infections with Haemonchus contortus and related gastrointestinal nematodes in small ruminants. Advances in Parasitology, 93, 239–351. https://doi.org/10.1016/bs.apar.2016.02.025 McRae, KM. et al. (2015). The host immune response to gastrointestinal nematode infection in sheep. Parasite Immunology, 37(12), 605-613. https://doi.org/10.1111/pim.12290. Mravcakova, D. et al. (2019). Natural chemotherapeutic alternatives for controlling of haemonchosis in sheep. BMC Veterinary Research, 15, 1-13. https://doi.org/10.1186/s12917-019-2050-2 Rouatbi, M. et al. (2016). Effect of the infection with the nematode Haemonchus contortus (Strongylida: Trichostrongylidae) on the haematological, biochemical, clinical and reproductive traits in rams. The Onderstepoort Journal of Veterinary Research, 83(1), 1-8. http://dx.doi.org/10.4102/ojvr.v83i1.1129 Varadyova, Z. et al. (2018). Effects of herbal nutraceuticals and/or zinc against Haemonchus contortus in lambs experimentally infected. BMC Veterinary Research, 14(1), 78. https://doi.org/10.1186/s12917-018-1405-4 Villalba, JJ. Et al. (2017). Phytochemicals in animal health: Diet selection and trade-offs between costs and benefits. Proceedings of the Nutrition Society, 76(2), 113-121. https://doi.org/10.1017/S0029665116000719

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

Background: This study aimed to investigate the effect of biologically active compounds (BAC) of Coleus amboinicus Lour. (CAL) herb fed to growing lambs on ruminal methane production, ruminal biohydrogenation of unsaturated fatty acids and meat characteristics. An in vitro trial (Experiment 1) comprising of control and three experimental diets (CAL constituting 10%, 15%, and 20% of the total diet) was conducted to determine an effective dose for in vivo experiments. After the in vitro trial, two in vivo experiments were conducted on six growing, rumen-cannulated lambs (Experiment 2) and 16 growing lambs (Experiment 3), which were assigned into the control (CON) and one experimental diet (20% of CAL). Several parameters were examined in vitro (pH, ammonia and VFA concentrations, protozoa, methanogens and select bacteria populations) and in vivo (methane production, digestibility, ruminal microorganism populations, meat quality, fatty acids profiles in rumen fluid and meat, transcript expression of 5 genes in meat). Results: The CAL lowered in vitro methane production by 51%. In the in vivo experiments, lambs fed CAL decreased methane production by 20% compared with the CON animals (Experiment 3), which corresponded to the reduced total methanogens counts in all experiments up to 28%, notably Methanobacteriales. In Experiment 3, CAL increased or tended to increase the numbers of Ruminococcus albus, Megasphaeraelsdenii, Butyrivibrioproteoclasticus, and Butyrivibriofibrisolvens. Dietary CAL suppressed the Holotricha population, but increased or tended to increase Entodiniomorpha population in Experiments 2 and 3. An increase in the polyunsaturated fatty acid (PUFA) proportion in the rumen of lambs was noted in response to the CAL diet, which was mainly attributable to the increase in C18:3 cis-9 cis-12 cis-15 (LNA) proportion. The CAL reduced the mRNA expressions of four investigated genes in meat (fatty acid synthase, stearoyl-CoA desaturase, lipoprotein lipase, and fatty acid desaturase 1). Conclusions:Summarizing, polyphenols of CAL (20% in diet) origin can mitigate ruminal methane production by inhibiting the methanogens communities. Supplementation of CAL also provides favorable conditions in the rumen by modulating ruminal bacteria involved in fermentation and biohydrogenation of fatty acids. CAL elevated the LNA concentration, which led to improved meat quality through increased deposition of n-3 PUFA.

Coleus amboinicus Lour., Lamiaceae, is a perennial herb that is native to Indonesia and also cultivated in Africa, Asia and Australia. The major phytochemicals responsible for its bioactivity are rosmarinic acid (RA) and its analogues, flavonoids and abietane diterpenoids. The possibility of cultivation in a colder climate would extend the use of this herb and provide new opportunities to herb growers and livestock farmers. Our study to compare feed value and phytochemical composition of C. amboinicus plants cultivated in its original region, Indonesia, and in Poland. The crude protein content was significantly higher in plants cultivated in Indonesia compared to those cultivated in Poland—21% and 13% of dry matter, respectively. The higher ADF contents were detected in C. amboinicus cultivated in Indonesia, 38–41%, in comparison to 34% in plants cultivated in Poland. The phytochemical composition was also significantly influenced by the cultivation location. Polish samples were higher in polyphenols (RA and its analogues), and also had 1.5–2-fold higher antioxidant potential, as measured by DPPH scavenging, phosphomolybdenum reduction and Fenton reaction driven lipid peroxidation. The Indonesian samples contained more diterpenoid compounds, such as dihydroxyroyleanone, and the sum of terpenoids was ca. 10 times higher than in samples from Poland (15.59–23.64 vs. 1.87 µg/g of extracts). In conclusion, C. amboinicus is suitable for cultivation in non-optimal climatic conditions but some nutritional properties and bioactivity are significantly affected.

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 &lt, 0.05), and the total antioxidant capacity was higher in MAL (p &lt, 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 &lt, 0.01 and p &lt, 0.05). Plasma cell numbers were higher in ARTMAL than the number in MAL (p &lt, 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.

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.

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.

Background: The aim of this study was to investigate the effect of diets containing organic zinc and a mixture of medicinal herbs on rumen microbial fermentation and histopathology in lambs.Methods: 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 mix of dry medicinal herbs (Herbs, 100 g 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). 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 days of the experiment. The rumen content was used to determine the fermentation parameters in vitro and in vivo and to quantify the microbes by molecular and microscopic methods. Samples of fresh rumen tissue were used for histopathological evaluation.Results: Quantitative analyses of the bioactive compounds 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 in FO, MS, AA and MC, respectively. The alkaloids content in FO was 6.015 g/kg DM. The effect of diets on values of total gas, methane and n-butyrate in vitro was observed (P < 0.046 and P < 0.001, respectively). Relative quantification by real-time PCR revealed a decrease in the total rumen bacterial population in the lambs in the Zn and Zn+Herbs groups (P < 0.05). A shift in the relative abundance of Ruminococcus albus, Streptococcus bovis, Butyrivibrio proteoclasticus and R. flavefaciens was observed in the Zn group. Morphological observation showed focal mixed infiltration of inflammatory cells within the lamina propria of the rumen in the Zn+Herbs group. Conclusion: The effect of the organic zinc and the herbal mix on rumen fermentation parameters in vitro was not confirmed in vivo because the rumen microbiota of lambs probably adapted to the zinc-supplemented diets. However, long-term supplementation of a diet combining zinc and medicinal herbs can diminish the health of the ruminal epithelium of lambs.

Paulownia is a genus of fast-growing trees that generate a huge mass of leaves, which can be utilized as a feed resource for ruminants. In the present study, the chemical and phytochemical composition, in vitro ruminal fermentation and methane production, and nutrient degradability of fresh Paulownia hybrid leaves (PL) and its silage (PLS) were investigated. The crude protein content of PL and PLS ranged from 132 to 199 g/kg dry matter, which was comparable to alfalfa silage (AS). Ensiling of PL increased the amount of both phenolic acids and flavonoids. The amino acid content increased, and total saturated fatty acid concentration tended to increase in PLS compared with PL. In the in vitro study with the ruminal fluid (batch culture), PL and PLS decreased pH (P

The aim of this study was to determine the effect of dietary supplementation with a mixture of selected medicinal herbs (Herbmix) on hematological and parasitological parameters and on the inflammatory response of lambs infected experimentally with the gastrointestinal parasitic nematode Haemonchus contortus . Herbmix contained selected medicinal herbs common in central Europe (8.55% each of Althaea officinalis , Petasites hybridus , Inula helenium , Malva sylvestris , Chamomilla recutita , Plantago lanceolata , Rosmarinus officinalis , Solidago virgaurea , Fumaria officinalis , Hyssopus officinalis and Melissa officinalis , 5% Foeniculum vulgare and 1% Artemisia absinthium ). Herbmix (100 g dry matter (DM)/d) was added to the basal diets of meadow hay (600 g DM/d) and a commercial concentrate (350 g DM/d) per animal. Twenty-four female lambs were divided into four groups: infected animals (Inf), infected animals supplemented with Herbmix (Inf + Herb), uninfected control animals (C) and uninfected animals supplemented with Herbmix (C + Herb). Inf and Inf + Herb lambs were infected orally with approximately 5000 L3 larvae of a strain of H. contortus susceptible to anthelmintics (MHco1). Herbmix supplementation for Inf + Herb and C + Herb began on day 0 (D0). Feces were collected and eggs per gram (EPG) of feces were quantified on D0, D20, D32, D50 and D60. Variables differed after 24 h of ruminal incubation of the individual medicinal herbs ( P 0.001), but the herbs had no negative effect on the fermentation patterns. The mean EPG counts were significantly lower by 45% between D32 and D50 ( P 0.05) and by 63% between D32 and D60 ( P 0.01) for the infected lambs receiving Herbmix (Inf + Herb). These EPG reductions were only 10 and 30% in infected group (Inf, P > 0.05). Mean cumulative gain in live weight was higher in Inf + Herb as compared to Inf by D32 ( P 0.01) and D50 ( P 0.001). The evaluation of red blood cells identified clinical signs of haemonchosis, such as anemia, in Inf and Inf + Herb. Total antioxidant capacity ( P 0.008) and activity of glutathione peroxidase ( P 0.005) in serum at D50 were lower in Inf and Inf + Herb than C and C + Herb. The concentration of serum calprotectin differed significantly ( P 0.001) in Inf vs. Inf + Herb. The Zn and Fe serum concentrations differed significantly ( P 0.05 and P 0.001, respectively) in Inf vs. Inf + Herb, Inf vs. C and Inf vs. C + Herb by D32. The mixture of the medicinal herbs thus contributed to slowing the dynamics of infection and improving the production indicators of the animals.

Simple Summary The gastrointestinal parasitic nematode Haemonchus contortus of small ruminants is an important target for chemoprophylaxis. Repeated use of anthelmintics in the form of synthetic drugs increases the risk of residues in food products and the development of anthelmintic resistance. However, the use of combinations of dry traditional medicinal plants as nutraceuticals is an alternative to chemotherapeutics for controlling haemonchosis in ruminants. Therefore, the aim of this study is to determine the effect of dietary supplementation with wormwood, mallow and their mix on parasitological status and inflammatory response in lambs experimentally infected with H. contortus. Simultaneously, the present study evaluated by the egg hatch test the in vitro anthelminthic effects of different concentrations (50–1.563 mg/mL) of the aqueous extracts of these plants. Our results revealed that the strong anthelmintic effect of both medicinal plants observed in vitro was not fully confirmed in vivo. This knowledge builds on our previously published findings and highlights that the effect of dry medicinal plants depends on the variety and synergy of plant polyphenols and the combination of bioactive compounds that together have an effect and contribute to a certain pharmacological efficacy. Abstract 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.

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 .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.

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 0.05). 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.

Two 24 h in vitro batch culture experiments were conducted to investigate the effects of fumarate addition (10 mmol L−1) on the ruminal fermentation parameters of selected medicinal herbs, and the effects of different doses of fumarate (0, 10, or 30 mmol L−1) on ruminal metabolism of organic acids when a high-concentrate diet (meadow hay and barley grain, 400/600, w/w) was supplemented with a mix of medicinal herbs (Artemisia absinthium L., Melissa officinalis L., Malva sylvestris L., Matricaria chamomilla L., Plantago lanceolata L., Foeniculum vulgare Mill., and Althaea officinalis L.). Depending on the concentration, fumarate treatment decreased methane production (by 10–11%) and increased propionate proportions (by 5–13%) with high-concentrate diets. The organic acid (fumaric, succinic, malic, and lactic acid) concentrations in the batch culture were measured at intervals of 0, 4, 6, 12, and 24 h. The time and organic acid concentrations with 10 mmol L−1 fumarate were well correlated (R 2 = 0.846). The amount of succinate was accumulated and metabolized more slowly than that of fumarate (>24 h). The addition of fumarate and a herb mix could positively influence in vitro ruminal fermentation parameters of high-concentrate diets by increasing the levels of propionate and succinate as well as the pH, and by decreasing of methane emissions.

Recently, the narrow-leafed, blue lupine (Lupinus angustifolius) seeds, including the Tango variety, have been used as a potential feed of livestock animals. However, there is still a limited knowledge of possible action of the Tango lupine seed meal (LSM) on ruminal fermentation, enteric methane production, milk yield and composition in dairy cattle. Thus, the present study aimed to investigate the effect of LSM supplementation to dairy cows on ruminal enteric methane production, microbial population, ruminal biohydrogenation and fatty acid (FA) proportion in the ruminal fluid and milk. An in vitro trial comprising of control and five experimental groups (supplementation of LSM at 20, 40, 60, 80, and 100 g/kg diet) was conducted to determine an effective dose level for an in vivo experiment. The in vitro trial was followed by an in vivo experiment conducted on 60 high-yielding Polish Holstein Friesian dairy cows assigned into two groups: the control (n = 30) and the experimental LSM group (n = 30; 2 kg LSM/day/cow). Supplementation of LSM decreased methane emission and archaeal abundances in vitro and in vivo and increased ammonia level in vivo. Ruminal total volatile fatty acid concentration and acetate proportion were reduced by LSM supplementation both in vitro and in vivo. Milk yield and its basic composition were not influenced by the LSM. However, the LSM increased the unsaturated FA content and decreased n6 to n3 FA ratio in milk. Supplementation of LSM upregulated the mRNA expressions of FASN and ELOVL5 genes and downregulated the LPL gene expression in milk somatic cells. The daily cost of LSM diet and the cost of 1 kg of milk production were lower compared with the control. It is concluded that LSM supplementation may improve milk quality without affecting milk yield and has the potential to decrease methane production in dairy cows.

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 0.05). The diets containing medicinal plants and organic zinc thus helped to modulate the characteristics of fermentation in ruminants.

a b s t r a c t The aim of this study was to determine the effects of a medicinal plant mixture (MP), sunflower oil (SO) and a combination of the medicinal plant mixture and sunflower oil (MPSO) on fermentation end products, the fatty acid composition of the rumen fluid, the ruminal microbial population as well as antioxidant status in the blood of sheep fed a high-concentrate diet. Four rumen-fistulated rams were randomly assigned according to a 4 × 4 Latin square design and fed a basal diet consisting of 720 g DM/day meadow hay and 540 g DM/day barley grain. The basal diet was supplemented with no additive (control), a medicinal plant mixture (MP, 10% replacement of meadow hay; 72 g DM/day), sunflower oil (SO, 36 g/day) and the combination (MPSO). Before the in vivo experiment, 24 h ruminal incubations in vitro were performed (meadow hay/barley grain; 400/600, w/w; MP; 10% replacement of meadow hay; SO; 3.5% DM; MPSO). The in vitro experiment with MP and MPSO increased the in vitro dry matter digestibility (P < 0.001) and decreased methane production (P = 0.021), while the MPSO treatment increased the concentration of polyunsaturated fatty acids (PUFA; P < 0.001). In sheep, the rumen characteristics were not affected by the treatments. No treatment effect on the rumen eubacterial population was detected by the 16S-polymerase chain reaction-denaturizing gradient gel electrophoresis method. The beneficial effects of MP and MPSO on PUFA concentration observed in vitro were not fully confirmed in vivo. However, these results could point to the promising effects of using a medicinal plant mixture in high-concentrate diets of ruminants without adverse effects on the fermentation characteristics and microbial ecosystem in sheep rumen.

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.

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

Aims Enrichment of wheat bran (WB), corn meal (CM) and barley flakes (BF) with the oleaginous fungus Cunninghamella echinulata (CE) might lead to effective use of these by-products in ruminant nutrition. We examined their effects on rumen fermentation and lipid metabolism. Methods and Results WB, CM and BF substrates without or with brewer's grains (WBG, CMG, BFG) and enriched with CE were incubated with meadow hay (MH, 500 : 500, w/w) in rumen fluid in vitro for 24 h. The dry matter of the CE-enriched substrates increased (by 2–4%); however, digestibility decreased (P

The impact of 3 months of daily administration of equimolar doses of inorganic and organic Selenium (Se) on the fatty acid composition of rumen contents and rumen bacteria and protozoa was studied. Twelve sheep were fed a basal diet of meadow hay (1380 g dry matter/sheep) and ground barley grain (450 g dry matter/sheep) containing a background Se concentration of 0.16 mg/kg DM. The sheep were randomly divided into three groups and fed the basal diet only (Control) or identical diets supplemented with inorganic (Na2SeO3; ISe) or organic Se (selenized yeast; OSe) in equimolar amounts of Se of 0.4 mg/kg of feed DM. The fatty acid (FA) profile was determined in the total rumen contents (TRC), bacterial fraction (BF) and ciliated protozoal fraction (PF). In the TRC, supplementation with OSe decreased the concentration of C18:2 cis-9, trans-11 (P

The effects of two probiotic inoculants Enterococcus faecium 2/3s (EF2/3s) and E. faecium 26/42 (EF26/42) on nutritional patterns and fatty acid concentration in corn silages were examined. Both inoculants were isolated and characterized at Institute of Animal Physiology Slovak Academy of Sciences. The whole crop corn (341 g of dry matter (DM) kg−1) was conserved at 21°C for 111 days during the three treatments: (1) corn without inoculant (CS, control), (2) corn inoculated by the strain EF2/3s (CS + EF2/3s) and (3) corn inoculated by the strain EF26/42 (CS + EF26/42). Counts of the inoculated strains were enumerated (5.58 and 4.92 log10 cfu g−1 for EF26/42 and EF2/3s, respectively) immediately after their inoculation. At the end of conservation, EF26/42 achieved 1.60 ± 0.26 log10 cfu g−1 and EF2/3s less than 1.0 log10 cfu g−1. Compared to CS, the lactate values of inoculated CS were greater (P < 0.001). It can be concluded that both bacterial inoculants effectively enhanced in vitro DM degradability and l...

Effects of diets supplemented with fruit seed oils on fermentation parameters, ciliated protozoan population, and fatty acid composition of the rumen fluid of dairy cows were studied in 24 h batch cultures. Two diets, one containing lucerne plus wheat meal (60:40%) and the other containing of meadow hay plus wheat meal (60:40%), were supplemented with either grape oil or black currant oil (50 g . kg -1 of dry matter). The control diet contained no oil supplementation. The oils were selected due to high content of linoleic acid (grape oil, 696 g . kg -1 of fatty acids; black currant oil, 586 g . kg -1 of fatty acids). Oil supplements did not affect the basal parameters of rumen fermentation. Interactions between the diets and oil supplements affected rumen methane production and either the total or the majority of rumen ciliate species examined. Although the diets had no effect on the total content of volatile fatty acids, the proportions of n-butyrate and iso-valerate were significantly affected. The concentration of polyunsaturated fatty acids was higher in the meadow hay diet than in the lucerne diet, whereas addition of oils increased polyunsaturated fatty acids content. Black currant oil supplementation proved to be more efficient in enhancing polyunsaturated fatty acids content in rumen fluid when compared to grape oil. In conclusion, both oil supplements considerably decreased methane production when lucerne was used as the diet, what could be the effect of detrimental influence of the type of diet and oil supplement on protozoa population. However, the supplements did not negatively affect other rumen parameters, hence may be considered as valuable supplements in ruminant nutrition.

We examined fermentation capacity of fecal microbial inocula of Przewalski horse ( Equus ferus przewalskii ), Asian wild ass - kulan ( Equus hemionus hemionus ), and Chapman zebra ( Equus quagga chapmani) in vitro. Interactions of the substrates (amorphous cellulose, wheat straw, meadow hay, xylan from oat spelt, and ground barley grain) and type of fecal inocula in the gas volume and in vitro dry matter digestibility were detected in all substrates after 72 hours of fermentation in five replicates for each substrate and type of inocula. No effects of fecal inocula sources were detected on total short-chain fatty acids concentrations. No live fecal ciliate population was present in kulan feces. Complex ciliate populations in zebra feces and the number and genera resembled ciliates from the colon of horses. Fresh feces of kulan and zebra were fractionated by galvanotaxis and centrifugation to separate fecal ciliates and bacteria. Specific activities (μmol of reducing equivalents/mL min mg protein) of carboxymethyl cellulase (CM-cellulase), xylanase, α-amylase, and inulinase were measured in crude cell-free extract of fecal ciliates (zebra), fecal bacteria (zebra and kulan), and total fecal preparation (zebra and kulan). All examined specific enzymatic activities were present in zebra fecal samples. We were unable to measure the inulinase activity and CM-cellulase activity in kulan fecal samples. Zebra ciliates are actively involved in the digestion of plant storage (α-amylase, 0.53 ± 0.02; inulinase, 1.77 ± 0.01, specific activities) and structural polysaccharides (CM-cellulase, 0.4 ± 0.15; xylanase, 0.26 ± 0.06). For the first time, we measured inulinase activity in intestinal ciliates.

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.