Oil-in-Water Nanoemulsion Can Modulate the Fermentation, Fatty Acid Accumulation, and the Microbial Population in Rumen Batch Cultures.

In this study, three oil-in-water nanoemulsions were tested in two stages: In the first stage, three levels (on the substrate dry matter (DM)), namely 3%, 6%, and 9%, of three different oils, olive oil (OO), corn oil (CO), and linseed oil (LO), in raw and nanoemulsified (N) forms were used separately in three consecutive rumen batch cultures trials. The second stage, which was based on the first stage's results, consisted of a batch culture trial that compared the raw and nanoemulsified (N) forms of all three oils together, provided at 3% of the DM. In the first stage, NOO, NCO, and NLO preserved higher unsaturated fatty acid (UFA) and less saturated fatty acid (SFA) compared to OO, CO, and LO, respectively; noticeably, NCO had UFA:SFA = 1.01, 1.16, and 1.34 compared to CO, which had UFA:SFA = 0.66, 0.69, and 0.72 when supplemented at 3%, 6%, 9% of DM, respectively. In the second stage, UFA:SFA = 1.04, 1.12, and 1.07 for NOO, NCO, NLO, as compared to UFA:SFA = 0.69, 0.68, and 0.72 for OO, CO, and LO supplemented at 3% of DM. In conclusion, oil-in-water nanoemulsions showed an ability to decrease the transformation of UFA to SFA in the biohydrogenation environment without affecting the rumen microorganisms.

Effects of raw and fermented rapeseed cake on ruminal fermentation, methane emission, and milk production in lactating dairy cows.

The objective of this study was to evaluate the effects of replacing raw rapeseed cake (RC) with fermented rapeseed cake (FRC) in the diet of dairy cows on methane (CH4) production, ruminal fermentation, and milk production, composition, and fatty acid composition. The Hohenheim gas test (exp. 1) was initially used to evaluate RC and FRC as substrates. Following batch fermentation, an in vitro study (exp. 2) was performed to assess the effects of replacing RC with FRC at 28.75, 57.5, 86.25, and 115 g/kg (FRC25, FRC50, FRC75, and FRC100) in the total mixed rations (TMR). Based on the in vitro results, the control TMR (115 g/kg dry matter (DM) of RC; CONRC) and experimental TMR (115 g/kg DM of FRC; FRC100) were chosen for an in vivo assessment. In exp. 3, four ruminally cannulated cows were used in a replicated 2 (group) × 2 (period) crossover design and fed the TMR ad libitum. In exp. 4, twenty multiparous Polish Holstein-Friesian cows in their mid-lactation (148 ± 26 d in milk) were used in a completely randomized design. The cows were fed a partial mixed ration without the RC and FRC, and the RC and FRC were supplied in a concentrate feeder at 2.65 kg/d/cow. The FRC100 markedly decreased CH4 production by 12% and archaeal population without adversely affecting nutrient digestibility. The molar proportion of propionate was increased, and the molar proportion of acetate and butyrate and acetate to propionate ratio were decreased by FRC100. No significant effects on milk production or composition, except an increase in milk urea concentration, were observed in cows fed FRC100. Milk C18:2 cis-9, trans-11 concentration was greater, and n-6 to n-3 fatty acid ratio was lower for FRC100 than CONRC. In-situ ruminal degradation of RC and FRC were explored using in-sacco techniques (exp. 5). The potential degradation and effective degradability of the DM, organic matter, and crude protein were significantly higher for FRC than RC. These results suggested that FRC could mitigate enteric CH4 production by decreasing archaeal abundances without adversely affecting milk production and ruminal fermentation in lactating cows.

Effect of Dried Apple Pomace (DAP) as a Feed Additive on Antioxidant System in the Rumen Fluid.

The study aimed to evaluate the effect of dried apple pomace (DAP) as a feed additive on the enzymatic activity and non-enzymatic compounds belonging to the antioxidant system in cattle rumen fluid. The experiment included 4 Polish Holstein−Friesian cannulated dairy cows and lasted 52 days. The control group was fed with the standard diet, while in the experimental group, 6% of the feedstuff was replaced by dried apple pomace. After the feeding period, ruminal fluid was collected. The spectrophotometric technique for the activity of lysosomal enzymes, the content of vitamin C, polyphenols, and the potential to scavenge the free DPPH radical was used. The enzyme immunoassay tests (ELISA) were used to establish the activity of antioxidants enzymes and MDA. Among the rumen aminopeptidases, a significant reduction (p < 0.01) from 164.00 to 142.00 was observed for leucyl-aminopeptidase. The activity of glycosidases was decreased for HEX (from 231.00 to 194.00) and ß-Glu (from 1294.00 to 1136.00), while a significant statistically increase was noticed for BGRD (from 31.10 to 42.40), α-Glu (from 245.00 to 327.00), and MAN (from 29.70 to 36.70). Furthermore, the activity of catalase and GSH (p < 0.01) was inhibited. In turn, the level of vitamin C (from 22.90 to 24.10) and MDA (from 0.36 to 0.45) was statistically higher (p < 0.01). The most positive correlations were observed between AlaAP and LeuAP (r = 0.897) in the aminopeptidases group and between ß-Gal and MAN (r = 0.880) in the glycosidases group. Furthermore, one of the most significant correlations were perceived between SOD and AlaAP (r = 0.505) and AcP (r = 0.450). The most negative correlation was noticed between α-Gal and DPPH (r = −0.533) based on these observations. Apple pomace as a feed additive has an influence on lysosomal degradation processes and modifies oxidation−reduction potential in the rumen fluid. Polyphenols and other low-weight antioxidant compounds are sufficient to maintain redox balance in the rumen.

Qualitative and Quantitative Analysis of Secondary Metabolites in Morphological Parts of Paulownia Clon In Vitro 112® and Their Anticoagulant Properties in Whole Human Blood.

It is not easy to find data in the scientific literature on the quantitative content of individual phytochemicals. It is possible to find groups of compounds and even individual compounds rather easily, but it is not known what their concentration is in cultivated or wild plants. Therefore, the subject of this study was to determine the content of individual compounds in the new Paulownia species, Oxytree, developed in a biotechnology laboratory in 2008 at La Mancha University in Spain. Six secondary metabolites were isolated, and their chemical structure was confirmed by spectral methods. An analytical method was developed, which was then used to determine the content of individual compounds in leaves, twigs, flowers and fruits of Paulownia Clon in Vitro 112®. No flavonoids were found in twigs and fruits of Oxytree, while the highest phenylethanoid glycosides were found in twigs. In this study, we also focused on biological properties (anticoagulant or procoagulant) of extract and four fractions (A-D) of different chemical composition from Paulownia Clon in Vitro 112 leaves using whole human blood. These properties were determined based on the thrombus-formation analysis system (T-TAS), which imitates in vivo conditions to assess whole blood thrombogenecity. We observed that three fractions (A, C and D) from leaves decrease AUC10 measured by T-TAS. In addition, fraction D rich in triterpenoids showed the strongest anticoagulant activity. However, in order to clarify the exact mechanism of action of the active substances present in this plant, studies closer to physiological conditions, i.e., in vivo studies, should be performed, which will also allow to determine the effects of their long-term effects.

Effect of Paulownia Leaves Extract Levels on In Vitro Ruminal Fermentation, Microbial Population, Methane Production, and Fatty Acid Biohydrogenation.

Paulownia is a fast-growing tree that produces a huge mass of leaves as waste that can be used as a feed source for ruminants. The previous study showed that phenolic compounds were the most active biological substances in Paulownia leaves, which affected the ruminal parameters and methane concentration. However, there are no scientific reports on the Paulownia leaves extract (PLE) containing phenolic compounds for their mode of action in the rumen. Phenolics constituted the main group of bioactive compounds in PLE (84.4 mg/g dry matter). PLE lowered the concentration of ammonia, modulated the VFA profile in the ruminal fluid, and decreased methane production. The PLE caused a significant reduction of in vitro dry matter degradability, reduced the number of methanogens and protozoa, and affected selected bacteria populations. PLE had a promising effect on the fatty acid profile in the ruminal fluid. Paulownia as a new dietary component or its extract as a feed additive may be used to mitigate ruminal methanogenesis, resulting in environmental protection and reducing ruminal biohydrogenation, improving milk and meat quality.

Effects of silybin supplementation on nutrient digestibility, hematological parameters, liver function indices, and liver-specific mi-RNA concentration in dogs.

BACKGROUND: Hepatopathies are an important group of disorders in dogs where proper nutritional care is crucial. Supplementation with a hepatoprotectant like silybin can improve liver function and should not interfere with nutrient digestibility. The purpose of this study was to investigate the effect of both pure silybin and commercial hepatoprotectant on nutrients digestibility, liver function indices and health status in healthy dogs (EXP1). Moreover, the second experiment (EXP2) investigated the effect of commercial hepatoprotectant on liver function tests and liver-associated miRNAs concentration in dogs with idiopathic liver disorder. RESULTS: Nutrient digestibility was not affected by treatment in EXP1. Supplementation did alter the serum fatty acid profile, with no clinical relevance. The levels of liver markers such as ALT, AST and GGT significantly decreased. In EXP2, supplementation with commercial hepatoprotectant containing silybin improved liver function tests. A decrease was observed in liver serum markers such as ALT, AST and miR122 concentration. CONCLUSIONS: EXP1 confirmed that silybin (whether pure or as a commercial hepatoprotectant) does not interfere with digestion which subsequently exerts no detrimental effect on dogs' health and metabolism. In EXP2, dietary supplementation with commercial hepatoprotectant containing silybin resulted in a decreased activity of serum liver markers, accompanied by a decrease in the concentration of liver-specific miRNA molecules. Liver function indices were consequently improved. Silybin supplementation can thus serve as an effective therapeutical tool in dogs with hepatopathies.

Mineral status and enteric methane production in dairy cows during different stages of lactation.

BACKGROUND: Lactating dairy cows are the greatest livestock contributor of methane, a major global greenhouse gas (GHG). However, good feeding management with adequate mineral intake can offers an effective approach to maintaining high levels of milk production and the health of dairy cows over the entire course of lactation, while also helping to reduce methane emission. The study described here investigated the plasma concentrations of both macroelements (Ca, Na, K, Mg, P) and microelements (Zn, Cu, Fe, Mn), as well as enteric methane emission and milk composition in high-yielding dairy cows in different lactation periods. The experiment was performed on Holstein-Friesian dairy cows with the average milk yield of 41 (± 9) L/day in a Polish commercial farm with modern dairy systems. A total of thirty high-yielding dairy cows were randomly assigned into three groups differing by lactation stage: early stage (Early, days 25-100), middle stage (Middle, days 101-250), and late stage (Late, day 250 and later). Dietary treatment for all cows was a total mixture ration (TMR) with maize and alfalfa silage the main forage components. RESULTS: The greatest milk yield and methane production were recorded in early-stage lactating cows, but the greatest methane intensity per kg of corrected milk was recorded in the late stage of lactation. Plasma concentrations of macroelements and microelements did not differ by lactation stages, but increased plasma concentrations of Zn and Fe and decreased plasma levels of Mg were noted during lactation. A positive correlation was found between plasma levels of Mg and other macroelements (Ca, Na, K), and between the concentrations of Fe and Zn, P in plasma, but no correlation between methane emission and mineral status was detected in the different lactation stages. CONCLUSIONS: Our results showed different mineral requirements and enteric methane emissions in each lactation stage. The feeding strategy and mineral utilization were adequate to maintain the health, mineral status, and milk production of the Holstein cows during the entire lactation period, and suggest an effective way of reducing methane emission.

Phytochemical Profile and Antioxidant Activities of Coleus amboinicus Lour. Cultivated in Indonesia and Poland.

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.

The effects of dietary medium-chain fatty acids on ruminal methanogenesis and fermentation in vitro and in vivo: A meta-analysis.

The efficacy of methane (CH4 ) suppression using medium-chain fatty acids (MCFA) remains inconclusive, despite a number of studies on this topic are available. We thus carried out a meta-analysis to integrate the published data on different concentrations and types of MCFA such as lauric acid and myristic acid, which investigated ruminal methanogenesis and fermentation in in vitro and in vivo experiments. In vitro MCFA sources were classified either as pure MCFA (lauric acid, myristic acid and their combinations) or as natural MCFA-rich oils (canola oil enriched with lauric acids, coconut oil, krabok oil and palm kernel oil). The MCFA sources used in the in vivo studies were coconut oil, lauric acid, myristic acid and the combination of lauric and myristic acids. A total of 41 studies (20 in vitro and 21 in vivo studies) were compiled in our database, which included the data on CH4 emission, digestibility, ruminal fermentation products and microbial populations. The results showed that the amount of CH4 production per unit of digested organic matter decreased linearly under in vitro conditions (p < .01) and tended to decrease quadratically under in vivo conditions (p < .07) with increasing doses of MCFA. Populations of protozoa (p < .01) in both in vitro and in vivo responded negatively in a linear manner, whereas Archaea population diminished quadratically (p = .04) only in the in vitro conditions with increasing doses of MCFA. Increasing dietary MCFA concentrations also reduced the fibre digestibility linearly (p < .05) in both in vitro and in vivo conditions. CH4 production for different sources of MCFA decreased in following order: coconut oil > lauric acid > myristic acid > mixed lauric and myristic acids > palm kernel oil > canola oil enriched with lauric acids > krabok oil. It can be concluded that the effect of MCFA on ruminal methanogenesis depends on the amount and type of MCFA.

Effects of various mastitis treatments on the reproductive performance of cows.

BACKGROUND: The purpose of the study described here was to evaluate the effects of different supportive treatments - such as antioxidants, immunomodulators, and nonsteroidal anti-inflammatory drugs (NSAIDs) - in mastitic cows treated with intramammary antibiotics on the efficacy of mastitis therapy and fertility indices. Fertility indices, including time to first insemination, conception rate, time between calving and conception (open days), and number of services per conception (insemination index), were evaluated for 300 dairy cows. Sixty cows without apparent clinical signs of mastitis were assigned 100 days after calving to a Control group. Another 240 cows with clinical mastitis were systematically divided into four experimental groups (I-IV) of 60 cows each. All mastitic cows were treated with approved intramammary antibiotics in recommended doses. Cows in Group I were treated with intramammary antibiotics only. Cows in Groups II, III, and IV, received intramammary antibiotic therapy and a single injection with antioxidants, an immunomodulator (lysozyme dimer), or an NSAID (flunixin meglumine), respectively. RESULTS: The lowest treatment efficacy of mastitic quarters and cows was noted in Group I (51.6 and 53.3%; p > 0.05). The best recovery rate was noted in Group II (63.3 and 66.7%; p > 0.05), followed by Group III (58.3 and 60.9%) and Group IV (58.3 and 58.0%; p > 0.05). The above data did not differ statistically (p > 0.05). The animals with mastitis (Groups I-IV) showed prolonged time to first insemination, more open days, higher insemination index, and lower conception rate than the control cows (p <  0.05). The conception rate of healthy cows and of successfully treated cows was insignificantly lower than that of cows required prolonged antibiotic therapy. Supportive treatments improved the mastitis recovery rate compared with intramammary antibiotics only. The efficacy of mastitis treatments affected the reproduction indices: in cows requiring prolonged treatment with antioxidants, a shorter time to first insemination was needed than in other groups (p <  0.05). Fewer days open were observed between the group with antioxidants and the control group (p <  0.05). CONCLUSIONS: Clinical mastitis negatively affects reproductive indices (days open, pregnancy rate after first AI, NSC) in dairy cows. Different types of supportive medicine, such as antioxidants (vitamin C and E, and ß-carotene), lysozyme dimer, or NSAID can be useful in improving fertility in mastitis cows treated with antibiotic only. It has been proven that each supportive treatment improved antibiotics efficiency and the antibiotic combined with the antioxidants was the most effective treatment.

Effects of feeding frequency and oil supplementation on feeding behavior, ruminal fermentation, digestibility, blood metabolites, and milk performance in late-lactation cows fed a high-forage diet.

Many dairy producers are keen to feed low-producing late-lactation cows only once per day (1×) to reduce production costs. This study examined effects of feeding frequency (FF: thrice versus once daily) on behavioral patterns, ruminal fermentation, and milk production performance of cows and supplementation of yellow grease oil (YO) rich in 18:2n-6 as a potential strategy to alleviate the possible negative effects of 1× daily feeding. Twenty-four late-lactation Holstein cows (215 ± 53.8 DIM) housed in tiestalls were assigned to 4 treatments according to a 2 × 2 factorial arrangement with 2 FF [3 times daily (3×) at 0800, 1400, and 2000 h; or 1× at 0800 h] and 2 high-forage total mixed rations (TMR), without (CON) or with 25 g/kg of dry matter of YO (YGO), in a randomized complete block design. Treatments were applied for 21 d. Feeding behavior was recorded every 5 min over a 24-h period on d 19. Fresh TMR and orts were sampled (d 15 to 21) and separated using a 3-screen (19, 8, and 1.18 mm) Penn State Particle Separator for sorting activity. Ruminal fluid samples were collected using oral stomach tubing on d 21. Cows on 1×-CON spent more time eating during the first 6 h after feeding at 0800 h than did cows on 3×-CON or 1×-YGO. Decreasing FF increased meal length and tended to increase meal size for CON cows, but supplementing YO increased meal bouts and reduced meal length and size for cows fed 1×. Cows on 1×-CON had the greatest ruminating time between 2000 and 0800 h compared with other treatments. Total and daytime distribution of lying time did not vary by treatments. Sorting activity was higher for cows fed frequently, and the extent of sorting was increased by oil supplementation. In the morning ruminal fluid samples, pH was not different among treatments, but in the evening samples 1× daily feeding reduced ruminal pH compared with 3×. In the morning and evening samples, ratios of acetate to propionate were the lowest for 1×-CON cows compared with other treatments. Dry matter intake and milk yield were similar among the groups. Milk fat content and yield decreased with 1×-CON treatment, but supplementing YO numerically increased milk fat for cows fed 1×. These results suggest that decreasing FF from 3× to 1× daily increases meal length, particularly after feed delivery, in cows fed high-forage diets; but supplementation of plant oil changes feeding patterns and may improve ruminal pH and milk fat in cows fed once a day.

Comparative Phytochemical, Antioxidant, and Hemostatic Studies of Extract and Four Fractions from Paulownia Clone in Vitro 112 Leaves in Human Plasma.

Background: The Paulownia Clone in Vitro 112, known as oxytree or oxygen tree, is a hybrid clone of the species Paulownia elongata and Paulownia fortunei (Paulowniaceae). The oxytree is a fast-growing hybrid cultivar that can adapt to wide variations in edaphic and climate conditions. In this work, Paulownia Clone in Vitro 112 leaves were separated into an extract and four fractions (A-D) differing in chemical content in order to investigate their chemical content using LC-MS analysis. The extract and fractions were also evaluated for their anticoagulant and antioxidant properties in a human plasma in vitro. Results: The Paulownia leaf extract contained mainly phenolic compounds (e.g., verbascoside), small amounts of iridoids (e.g., aucubin or 7-hydroxytometoside) and triterpenoids (e.g., maslinic acid) were also detected. Our results indicate that the extract and fractions have different effects on oxidative stress in human plasma treated with H2O2/Fe in vitro, which could be attributed to differences in their chemical content. For example, the extract and all the fractions, at the two highest concentrations of 10 and 50 µg/mL, significantly inhibited the plasma lipid peroxidation induced by H2O2/Fe. Fractions C and D, at all tested concentrations (1-50 µg/mL) were also found to protect plasma proteins against H2O2/Fe-induced carbonylation. The positive effects of fraction C and D were dependent on the dose. Conclusions: The extract and all four fractions, but particularly fractions C and D, which are rich in phenolic compounds, are novel sources of antioxidants, with an inhibitory effect on oxidative stress in human plasma in vitro. Additionally, the antioxidant potential of fraction D may be associated with triterpenoids.

The effect of total and individual alfalfa saponins on rumen methane production.

BACKGROUND: Ten varieties of alfalfa (Medicago sativa L.) were evaluated for saponin content. Two of the most promising varieties were chosen so that their effect on rumen fermentation and methane production could be studied. Initially, four Hohenheim gas tests (HGT) were performed to test the effect of increased levels of total saponin extracted from the two alfalfa cultivars (Kometa and Verko) - either as fresh material or ensiled - on the total bacteria, total protozoa, methane emission, and selected methanogenic population. Afterwards, seven particular saponins were extracted from fresh alfalfa of the Kometa variety and tested in 24 h batch fermentation culture experiments. RESULTS: The ensiled forms of both the Verko and Kometa alfalfa varieties seem to be good sources of saponin, capable of reducing methane production (P < 0.05) without negatively affecting the basic fermentation parameters. Of the two evaluated varieties, Kometa was the most effective, and the saponins extracted from its roots 3-Glc,28-Glc Ma, medicagenic saponin, and 3-Glu Ma showed the most evident effect (P = 0.0001). The most promising aerial alfalfa saponin in mitigating methane production was soysaponin I K salt (P = 0.0001). Three mixtures of saponins were tested and all were found to mitigate methane production; however, one mixture (MIX 1) did so only to a very small extent. CONCLUSION: Saponins have been observed to have differing effects depending on their source; however, the mode of action of saponins depends on their direct or probable indirect effect on the microorganisms involved in methane production. © 2019 Society of Chemical Industry.

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.

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.

Effects of berry seed residues on ruminal fermentation, methane concentration, milk production, and fatty acid proportions in the rumen and milk of dairy cows.

Strawberry (SB), black currant (BC), and raspberry seed (RB) residues were used in 3 experiments to study their effects on ruminal fermentation, methane concentration, and fatty acid (FA) proportions in the ruminal fluid and milk of dairy cows. Initially, a batch fermentation in vitro study (experiment 1) was performed to investigate the effects of the 3 berry residues on basic ruminal fermentation parameters. Total volatile fatty acid concentrations, including acetate, propionate, and butyrate, increased in the BC group compared with other treatments. Based on the preliminary in vitro results, 2 consecutive in vivo experiments were conducted using 4 Polish Holstein-Friesian cows fitted with rumen cannulas (experiment 2) and 30 lactating Polish Holstein-Friesian dairy cows (experiment 3) in a replicated 2 × 2 crossover design. Cows in both experiments received a partial mixed ration (PMR) in 2 variants: (1) a control diet of PMR + 2 kg of concentrate (control); (2) PMR + 2 kg of BC seed residues (BC). The BC diet did not mitigate ruminal methane production. Ruminal fermentation (experiment 2) was not affected by the BC diet; however, the concentrations of C18:1 trans-11 and C18:2 cis-9,trans-11 increased significantly by 91 and 131%, respectively. Likewise, concentrations of total trans C18:1 and total monounsaturated FA in ruminal fluid were increased significantly by BC seed residues. In experiment 3, BC significantly increased milk fat C18:1 trans-11, C18:2 cis-9,trans-11, n-3, n-6, and polyunsaturated FA concentrations without affecting milk production performance. In conclusion, the amount (2 kg/d) of BC used in this study did not adversely affect ruminal fermentation or milk production and composition. However, using BC increased proportions of unsaturated FA and conjugated linoleic acid in milk. Although dietary BC did not exert a strong methane inhibition effect, it could represent an inexpensive alternative concentrate to improve beneficial FA in milk without negative effects on rumen fermentation and production parameters in dairy cows. Incorporation of berry seed residues in diets would be profitable economically and nutritionally for dairy cattle production.

Structural and quantitative changes of saponins in fresh alfalfa compared to alfalfa silage.

BACKGROUND: Alfalfa (Medicago sativa) is an important legume used in animal feed. The primary objective of the present study was to analyze and trace the individual saponins in fresh and ensiled alfalfa of ten varieties, with the aim of evaluating the structural changes that saponins undergo during ensiling. A secondary objective was to examine whether any of the changes in saponin content were associated with changes in the basic nutrient composition of the ensiled alfalfa. RESULTS: The total saponin concentration increased when the fresh alfalfa was processed into silage. Three of the zanhic acid glycosides were degraded substantially, whereas the other two increased in all the tested varieties. Five of the seven medicagenic acid glycosides were not changed, whereas the remaining saponins completely disappeared. Finally, two glycosides of soyasapogenol B displayed an inverse relationship to each other. CONCLUSION: The nutrient content of fresh material and silages remained relatively constant, with some variations in the crude protein content of the selected alfalfa varieties. The total concentration of saponins increased two- to three-fold after ensilation over the levels present in fresh alfalfa material. The increases in saponin concentration showed a negative linear relationship between protein on a dry matter basis and saponin content. These findings are significant because no known published data have shown this transformation in the ensiling of alfalfa. © 2018 Society of Chemical Industry.

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 Effect of Different Levels of Cu, Zn and Mn Nanoparticles in Hen Turkey Diet on the Activity of Aminopeptidases.

The aim of the study was to estimate the influence of the different levels of Cu, Zn, and Mn nanoparticles on the activity of aminopeptidases in turkey. An experiment was carried out on 144 turkey hen Hybrid Converter. The birds were divided into groups given standard- and nanoparticle-supplementation of different level of copper (Cu 20, 10, 2 mg/kg), zinc (Zn 100, 50, 10 ppm), and manganese (Mn 100, 50, 10 ppm), covering respectively 100%, 50%, and 10% of the physiological demands for those minerals in the diet. The activity of aminopeptidases (alanyl: AlaAP, leucyl: LeuAP and arginyl: ArgAP) after supplementation of minerals was determined in the breast and thigh turkey muscle. The strongest effect of interaction among minerals supplementation form and dose on the activity levels of the aminopeptidases in thigh muscle was observed for nano-Cu already at the lowest dose of 2 mg/kg. In this dose (covering 10% of the birds' demand) nano form of supplementation significantly increased the activity of Ala-, Leu-, and ArgAP (877, 201, and 719, respectively), compared to standard form of supplementation (461, 90.5, and 576, respectively). In turn, in breast muscle, after supplementation covering 10% of the demand with the nano-Cu, nano-Zn, and nano-Mn compared to the standard form, we did not observe any significant difference in the activity levels of any of the investigated aminopeptidases, except for AlaAP under Zn supplementation. Supplementation with the 20 mg/kg of Nano-Cu (100% of demand) and with 10 mg/kg of Nano-Cu (50% of demand) inhibited the activity of all of the three aminopeptidases in thigh muscle. Supplementation of the minerals in nano form into the diet, especially of Cu and Zn in the dose covering 10% of the demand is relevant to maintain homeostasis in turkey muscles, as indicated by the activity of the aminopeptidases.

Multifactorial analysis of the follicular environment is predictive of oocyte morphology in cattle.

Numerous attempts have been recently made in the search for a reliable, fast and noninvasive assay for selection of oocytes suitable for in vitro embryo production. Potential markers have been described in the follicle such as follicular fluid (FF) or cumulus cells (CCs). However, the reported findings are contradictory, which may reflect the complexity of metabolism of the ovarian follicle. In the present experiment, a data set from individual follicles of known diameter was obtained: cumulus-oocyte complex (COC) morphology, fatty acid composition and glucose concentration in FF as well as apoptotic index in CCs. The obtained data was statistically analyzed either separately (univariate analysis) or simultaneously (multivariate analysis) to examine its predictive value in morphology assessment of bovine COCs. Although the univariate analysis yielded a complex relation system of the selected parameters, no clear outcome could be established. In multivariate analysis, the concentration of the four fatty acids (C16:0, C16:1, C18:1cis9, C22:5n3) and Δ(9)-desaturase (16) as well as elongase activities were selected as covariates. This allowed prediction of the morphology of a COC with an accuracy of 72%, which is the most interesting finding of the experiment. The present study indicates that the multifactorial model comprising of selected parameters related to the follicle appeared more effective in predicting the morphology of a bovine COC, which may improve the effectiveness of in vitro production systems.