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Bioactive Compounds, Ruminal Fermentation, and Anthelmintic Activity of Specialty Coffee and Spent Coffee Grounds In Vitro
We quantified the bioactive compounds of Ethiopian coffee (ETH), spent coffee grounds SCGs from ETH (SCG-ETH), and mixed SCGs (SCG-MIX) prepared by filtration methods and investigated the effect of SCG-ETH on ruminal fermentation as well as the anthelmintic activity of ETH. Three substrates, meadow hay (MH)-barley grain (MH-BG), MH-SCG-ETH, and BG-SCG-ETH (1:1 w/w), were fermented using an in vitro gas production technique. The bioactive compounds were quantitatively analyzed using ultra-high-resolution mass spectrometry. We performed an in vitro larval development test to determine the anthelmintic effect of an aqueous extract of ETH against the gastrointestinal nematode (GIN) Haemonchus contortus. The total content of bioactive compounds was highest in SCG-ETH, followed by SCG-MIX and ETH (35.2, 31.2, and 20.9 mg/g dry matter, respectively). Total gas and methane production (p < 0.001) were decreased by both MH-SCG-ETH and BG-SCG-ETH. The in vitro digestibility of dry matter was higher for MH-SCG-ETH and BG-SCG-ETH than for MH-BG. The aqueous ETH extract exhibited a strong larvicidal effect, with a mean lethal dose of 13.2 mg/mL for 50% mortality and 31.9 mg/L for 99% mortality. SCG substrates have the potential to modulate ruminal fermentation and serve as a source of anthelmintic bioactive compounds against GINs in ruminants.
The effect of ensiled paulownia leaves in a high-forage diet on ruminal fermentation, methane production, fatty acid composition, and milk production performance of dairy cows
Abstract Background The use of industrial by-products rich in bioactive compounds as animal feeds can reduce greenhouse gas production. Paulownia leaves silage (PLS) was supplemented to dairy cows' diet and evaluated in vitro (Exp. 1; Rusitec) and in vivo (Exp. 2, cannulated lactating dairy cows and Exp. 3, non-cannulated lactating dairy cows). The study investigated the PLS effect on ruminal fermentation, microbial populations, methane production and concentration, dry matter intake (DMI), and fatty acid (FA) proportions in ruminal fluid and milk. Results Several variables of the ruminal fluid were changed in response to the inclusion of PLS. In Exp. 1, the pH increased linearly and quadratically, whereas ammonia and total volatile fatty acid (VFA) concentrations increased linearly and cubically. A linear, quadratic, and cubical decrease in methane concentration was observed with increasing dose of the PLS. Exp. 2 revealed an increase in ruminal pH and ammonia concentrations, but no changes in total VFA concentration. Inclusion of PLS increased ruminal propionate (at 3 h and 6 h after feeding), isovalerate, and valerate concentrations. Addition of PLS also affected several populations of the analyzed microorganisms. The abundances of protozoa and bacteria were increased, whereas the abundance of archaea were decreased by PLS. Methane production decreased by 11% and 14% in PLS-fed cows compared to the control in Exp. 2 and 3, respectively. Exp. 3 revealed a reduction in the milk protein and lactose yield in the PLS-fed cows, but no effect on DMI and energy corrected milk yield. Also, the PLS diet affected the ruminal biohydrogenation process with an increased proportions of C18:3 cis-9 cis-12 cis-15, conjugated linoleic acid, C18:1 trans-11 FA, polyunsaturated fatty acids (PUFA), and reduced n6/n3 ratio and saturated fatty acids (SFA) proportion in milk. The relative transcript abundances of the 5 of 6 analyzed genes regulating FA metabolism increased. Conclusions The dietary PLS replacing the alfalfa silage at 60 g/kg diet can reduce the methane emission and improve milk quality with greater proportions of PUFA, including conjugated linoleic acid, and C18:1 trans-11 along with reduction of SFA. Graphical Abstract Graphical abstract of the experimental roadmap
Growth Performance and Ruminal Fermentation in Lambs with Endoparasites and In Vitro Effect of Medicinal Plants
We investigated growth performance and ruminal fermentation associated with gastrointestinal nematode (GIN) Haemonchus contortus in lambs and in vitro ruminal fermentation of mallow, chamomile, fumitory, wormwood (Herbmix), and chicory using inoculum from GIN-infected lambs. Twelve lambs were equally divided into two groups: uninfected animals (CON) and animals infected (INF) with approximately 5000 third-stage larvae derived from the MHCo1 strain of GIN H. contortus. Two lambs per group were killed on days 48, 49, and 50 after infection and ruminal content was collected separately from each lamb. Batch cultures of ruminal fluid from CON and INF were incubated for 24 h in vitro with 0.25 g meadow hay, Herbmix, and chicory using an in vitro gas production technique. Daily weight gain was relatively lower in the INF than the CON group, but not significantly (72.6 vs. 130.1 g/day). The ruminal populations of protozoa, bacteria, total Archaea, Methanobacteriales, and Methanomicrobiales were significantly higher in the INF than in the CON group. The substrates affected the concentrations of n-butyrate, iso-butyrate, n-valerate, iso-valerate, ammonia-N, total gas, and methane (p < 0.001) in vitro. GIN infection affected fermentation and microbial population in the rumens of the lambs, and chicory was a promising substrate to modulate ruminal fermentation in vitro.
Potential of Paulownia Leaves Silage in Lamb Diet to Improve Ruminal Fermentation and Fatty Acid Profile − An in vitro Study
Abstract Environmental impact, quality, and quantity of food products of ruminant origin (especially beef and mutton) are considered major challenges in meeting the nutritional requirements of the growing human population worldwide. Therefore, we conducted this in vitro study to explore the potential of Paulownia leaves silage to reduce the environmental impact of feedlot lamb production and improve ruminal fatty acids (FAs) profile by influencing ruminal biohydrogenation. In the present study, Paulownia leaves silage (PLS) and alfalfa silage (AAS) were mixed in a proportion of 1:0 (Control, PLS 0%), 0.75:0.25 (PLS 25%), 0.5:0.5 (PLS 50%), 0.25:0.75 (PLS 75%) and 0:1 (PLS 100%) on dry matter basis in the lamb diet. The experimental findings demonstrated that 100% replacement of AAS with PLS in the lamb diet significantly improved the ruminal fermentation by increasing the in vitro DM degradability (P<0.01), total volatile fatty acid (P<0.01), and propionate production (P<0.01) while reducing the acetate: propionate (A/P) ratio (P<0.01) and CH4 concentration (mM; L and Q P<0.05) and CH4 production, mM/g DM (L and Q P<0.05). Meanwhile, 100% PLS inclusion in the diet increased the total monounsaturated fatty acids (P<0.05), total unsaturated fatty acids (P<0.01) and total n-3 fatty acids (P<0.05) particularly alpha-linolenic acid (ALA; C18:3 n-3; P<0.05) in the rumen fluid after incubation. Moreover, the total saturated fatty acids concentration was reduced (P<0.01). These findings suggested that PLS could be a climate-friendly and sustainable alternative to AAS in the lamb feedlot diet for quality meat production.
Effect of Sainfoin (Onobrychis viciifolia) Pellets on Rumen Microbiome and Histopathology in Lambs Exposed to Gastrointestinal Nematodes
Our study analyzed the ruminal fermentation and microbiome, hematological profile, and abomasal histopathology of lambs experimentally infected with a gastrointestinal nematode (GIN) and fed sainfoin pellets (SFPs; 600 g DM/d/animal) for 14 d. Twenty-four lambs infected with Haemonchus contortus were divided into two separated groups: animals fed meadow hay (control) and animals fed SFPs. The ruminal contents, fermentation parameters, and microbiome in vitro and in vivo were determined using molecular and microscopic techniques. Ruminal contents in the SFP group indicated smaller populations of Archaea (p < 0.001), Methanomicrobiales (p = 0.009), and lower methane concentrations in vitro (p = 0.046) and in vivo (p = 0.030) than the control group. The relative abundance of Butyrivibrio fibrisolvens quantified by real-time PCR was higher in the lambs with the SFP diet (p = 0.05). Haemonchosis affected the number of red blood cells of the lambs (p < 0.001). The lambs in the SFP group had a higher percentage of damaged abomasa glands than did the control group (p = 0.004). The consumption of SFPs by GIN-infected lambs may affect ruminal methanogens and subsequently decrease methane emission without undesirable changes in the ruminal microbiome or the health of the animals.
Insights into the role of bioactive plants for lambs infected with Haemonchus contortus parasite
Bioactive plants provide therapeutic and prophylactic effects to ruminants. We determined the effect of grazing on natural meadow grassland enriched with experimentally sown chicory (Cichorium intybus) on parasitological status, pasture larval infectivity, antioxidant parameters, and the histology of abomasal tissue in lambs experimentally infected with the parasitic gastrointestinal nematode (GIN) Haemonchus contortus. We also qualitatively identified the main polyphenols in the meadow grassland and phenolic metabolites in the feces of the lambs. Sixteen lambs were orally infected with approximately 5,000 infective larvae (L3) of H. contortus. The lambs were divided into two groups: lambs grazing on a plot consisting exclusively of meadow pasture which serves as control group and lambs grazing on a plot where approximately 25% of a meadow grassland was reclaimed with chicory. The experimental period was 144 days. The number of eggs per gram (EPG) of feces was quantified on D21, D34, D48, D62, D76, D89, D103, D118, D131, and D144 post-infection. Pasture contamination with H. contortus L3 was examined. EPG in both groups of lambs was highest at D34. Egg shedding was significantly lower in both groups from D48 onwards, with a reduction of >95% from D103 onwards. Pasture contamination with L3 was highest at D41 but was then significantly lower in both groups. The total antioxidant capacity, the activity of glutathione peroxidase and the concentration of malondialdehyde in the serum changed significantly during the experiment (p < 0.003, < 0.001, and < 0.016, respectively). At least 54 species of meadow plants were identified on both pasture plots; plant bioactive compounds identified were mainly phenolic acids, flavonoids, and glucosides. Phenolic metabolites (e.g., coumaric acid, chicory acid, salvigenin, and esters of gallic acid) were identified in the feces of the lambs. In some lambs, the morphological observation identified small histopathological changes in the abomasal tissues typical of hemonchosis. Both the natural meadow pasture and the pasture enriched with experimentally sown chicory slowed the dynamics of GIN infection and pasture contamination with L3 by mobilizing the antioxidant defensive system and gradually increasing the resistance of the infected lambs, probably due to the beneficial effects of plant bioactive substances.
Changes in Growth and Metabolic Profile of Scutellaria baicalensis Georgi in Response to Sodium Chloride
Scutellaria baicalensis Georgi is a valuable medicinal plant of the Lamiaceae family. Its roots have been used in Traditional Chinese Medicine (under the name Huang-qin) since antiquity and are nowadays included in Chinese and European Pharmacopoeias. It is abundant in bioactive compounds which constitute up to 20% of dried root mass. These substances are lipophilic flavones with unsubstituted B-ring, baicalein, and wogonin and their respective glucuronides–baicalin and wogonoside being the most abundant. The content of these compounds is variable and the environmental factors causing this remain partially unknown. The role of these compounds in stress response is still being investigated and in our efforts to measure the effect of NaCl treatment on S. baicalensis growth and metabolic profile, we hope to contribute to this research. Short-term exposure to salt stress (50, 100, and 150 mM NaCl) resulted in a marked increase of baicalein from 1.55 mg to 2.55 mg/g DM (1.6-fold), baicalin from 8.2 mg to 14.7 mg (1.8-fold), wogonin from 4.9 to 6.8 (1.4-fold), and wogonoside from 3.3 to 6.8 mg/g DM (2-fold) in the roots. Conversely, in the aerial parts, the content of individual major flavonoids: carthamidine-7-O-glucuronide and scutellarein-7-O-glucuronide decreased the most by 10–50% from 18.6 mg to 11.3 mg/g (1.6-fold less) and from 6.5 mg to 3.4 mg/g DM (0.52-fold less), respectively. The amino acid profile was also altered with an increase in root concentrations of the following amino acids: arginine from 0.19 to 0.33 mg/g (1.7-fold), glutamate from 0.09 to 0.16 mg/g DM (1.6-fold), alanine from 0.009 to 0.06 mg/g (6.8-fold), proline from 0.011 to 0.029 (2.4-fold) and lysine from 0.016 to 0.063 mg/g (3.9-fold). Aspartate concentration decreased from 0.01 to 0.002 mg/g (4.8-fold less) at 150 mM NaCl. In the aerial parts, the concentration and variation in levels of specific amino acids differed among groups. For instance, the glutamate content exhibited a significant increase exclusively in the treatment group, rising from 0.031 to 0.034 mg/g, representing a 1.2-fold increase. Proline concentration showed a marked increase across all treated groups with the highest from 0.011 to 0.11 mg/g (10-fold). In conclusion, moderate salt stress was shown to increase S. baicalensis root biomass and flavonoid content which is rarely observed in a glycophyte species and provides a foundation for further studies on the mechanisms of osmotic stress adaptation on the specialized metabolism level.
Dietary Coleus amboinicus Lour. decreases ruminal methanogenesis and biohydrogenation, and improves meat quality and fatty acid composition in longissimus thoracis muscle of lambs
Abstract Background Methane production and fatty acids (FA) biohydrogenation in the rumen are two main constraints in ruminant production causing environmental burden and reducing food product quality. Rumen functions can be modulated by the biologically active compounds (BACs) of plant origins as shown in several studies e.g. reduction in methane emission, modulation of FA composition with positive impact on the ruminant products. Coleus amboinicus Lour. (CAL) contains high concentration of polyphenols that may potentially reduce methane production and modulate ruminal biohydrogenation of unsaturated FA. This study aimed to investigate the effect of BAC of Coleus amboinicus Lour. (CAL) fed to growing lambs on ruminal methane production, biohydrogenation of unsaturated FA and meat characteristics. In this study, the in vitro experiment aiming at determining the most effective CAL dose for in vivo experiments was followed by two in vivo experiments in rumen-cannulated rams and growing lambs. Experiment 1 (RUSITEC) comprised of control and three experimental diets differing in CAL content (10%, 15%, and 20% of the total diet). The two in vivo experiments were conducted on six growing, rumen-cannulated lambs (Exp. 2) and 16 growing lambs (Exp. 3). Animals were assigned into the control (CON) and experimental (20% of CAL) groups. 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 CAL lowered in vitro methane production by 51%. In the in vivo Exp. 3, CAL decreased methane production by 20% compared with the CON group, which corresponded to reduction of total methanogen counts by up to 28% in all experiments, notably Methanobacteriales. In Exp. 3, CAL increased or tended to increase populations of some rumen bacteria (Ruminococcus albus, Megasphaera elsdenii, Butyrivibrio proteoclasticus, and Butyrivibrio fibrisolvens). Dietary CAL suppressed the Holotricha population, but increased or tended to increase Entodiniomorpha population in vivo. 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. CAL reduced the mRNA expression of four out of five genes investigated in meat (fatty acid synthase, stearoyl-CoA desaturase, lipoprotein lipase, and fatty acid desaturase 1). Conclusions Summarizing, polyphenols of CAL origin (20% in diet) mitigated ruminal methane production by inhibiting the methanogen communities. CAL supplementation also improved ruminal environment by modulating ruminal bacteria involved in fermentation and biohydrogenation of FA. Besides, CAL elevated the LNA concentration, which improved meat quality through increased deposition of n-3 PUFA.
Pulsed Electric Field (PEF) Treatment Results in Growth Promotion, Main Flavonoids Extraction, and Phytochemical Profile Modulation of Scutellaria baicalensis Georgi Roots
This study aims to explore the effect of pulsed electric field (PEF) treatment as a method very likely to result in reversible electroporation of Scutellaria baicalensis Georgi underground organs, resulting in increased mass transfer and secondary metabolites leakage. PEF treatment with previously established empirically tailored parameters [E = 0.3 kV/cm (U = 3 kV, d = 10 cm), t = 50 µs, N = 33 f = 1 Hz] was applied 1–3 times to S. baicalensis roots submerged in four different Natural Deep Eutectic Solvents (NADES) media (1—choline chloride/xylose (1:2) + 30% water, 2—choline chloride/glucose (1:2) + 30% water, 3—choline chloride/ethylene glycol (1:2), and 4—tap water (EC = 0.7 mS/cm). Confocal microscopy was utilized to visualize the impact of PEF treatment on the root cells in situ. As a result of plant cell membrane permeabilization, an extract containing major active metabolites was successfully acquired in most media, achieving the best results using medium 1 and repeating the PEF treatment twice (baicalein <LOQ, baicalin 12.85 µg/mL, wogonin 2.15 µg/mL, and wogonoside 3.01 µg/mL). Wogonin concentration in NADES media was on par with the control (plants harvested on the day of the experiment, ultrasound-mediated methanolic extraction, Cwogonin = 2.15 µg/mL). After successful extraction, PEF treatment allowed the plants to continue growing, with the lowest survival rate across treated groups being 60%. Additionally, an enhancement in plant growth parameters (length and fresh mass of the roots) and significant changes in the S. baicalensis root phytochemical profile were also observed.
Potential of chicory (Cichorium intybus) sward to improve growth performance and the fatty acid profile of rumen fluid, liver, muscle, and subcutaneous fat tissues of lamb
From Waste to Feed: Enhancing Sustainability in Ruminant Nutrition Through Cashew Apple Utilization – A Review
Abstract Growing global demand for livestock products has intensified the need for sustainable feed resources in ruminant nutrition, particularly in developing regions. With feed costs comprising up to 75% of production expenses, exploring alternative feed sources can offer both economic and environmental benefits. This study explored the potential of cashew apple (CA) as a novel and sustainable by-product for ruminants by reviewing relevant articles from databases such as Scopus, Web of Science, PubMed, and Google Scholar. CA has an average proximate composition (dry matter basis) of 130 g/kg crude protein, 71.7 g/kg crude fibre, 61.2 g/kg ether extract, 49.3 g/kg ash, 640 g/kg nitrogen free extract, and 2973 kcal/kg metabolizable energy, making it a valuable addition to ruminant diets. It can reduce reliance on traditional and high-cost feed sources and mitigate environmental pollution by reducing waste and contributing to circular agricultural practices. However, limitations such as high moisture content in fresh cashew fruit, seasonal availability, and rapid spoilage necessitate effective processing methods. There is a knowledge gap on the use of CA in ruminant diets, and there is a need to assess its impact on ruminal pH, short-chain fatty acids production, ammonia production, other rumen fermentation profiles, and animal performance. Based on the quantitative data of this study about the proximate composition of CA and some animal studies, it could be a viable and eco-friendly supplement to the ruminant diets. Addressing its storage and preservation challenges can unlock its potential in diversified, eco-friendly ruminant systems.