Chemical Composition and Nutritive Value of Almond Hulls from Two Almond Varieties and Influence of Including Almond Hulls in the Diet on In Vitro Ruminal Fermentation and Methane Production.

Almond hulls (AH) are frequently used in dairy ruminant feeding, but information on variability of their nutritive value and their potential effects on CH4 production is still scarce. The influence of almond variety (Guara vs. Soleta) on chemical composition and energy value of AH was investigated using 10 samples per variety collected in 2 consecutive years. Guara-AH had greater (p ≤ 0.015) ash, protein, and fat content, but lower (p ≤ 0.001) fiber than Soleta-AH. The metabolizable energy content estimated from chemical composition and in vitro gas production was 8.5% greater for Guara than for Soleta samples. Harvesting year significantly affected most of the chemical fractions. The in vitro ruminal fermentation of diets for dairy ruminants including increasing amounts of dried AH (8, 16 and 24% of the total diet; fresh matter basis) indicated that AH can be included up to 16% of the diet, partially substituting corn, wheat bran and sugar beet pulp without detrimental effects on in vitro volatile fatty acid (VFA) production. In contrast, when AH replaced alfalfa hay and corn, VFA production was reduced at all levels of AH inclusion. No antimethanogenic effects of AH were detected in the in vitro incubations.

Effects of Garlic Oil and Cinnamaldehyde on Sheep Rumen Fermentation and Microbial Populations in Rusitec Fermenters in Two Different Sampling Periods.

Garlic oil (GO) and cinnamaldehyde (CIN) have shown potential to modify rumen fermentation. The aim of this study was to assess the effects of GO and CIN on rumen fermentation, microbial protein synthesis (MPS), and microbial populations in Rusitec fermenters fed a mixed diet (50:50 forage/concentrate), as well as whether these effects were maintained over time. Six fermenters were used in two 15-day incubation runs. Within each run, two fermenters received no additive, 180 mg/L of GO, or 180 mg/L of CIN. Rumen fermentation parameters were assessed in two periods (P1 and P2), and microbial populations were studied after each of these periods. Garlic oil reduced the acetate/propionate ratio and methane production (p < 0.001) in P1 and P2 and decreased protozoal DNA concentration and the relative abundance of fungi and archaea after P1 (p < 0.05). Cinnamaldehyde increased bacterial diversity (p < 0.01) and modified the structure of bacterial communities after P1, decreased bacterial DNA concentration after P2 (p < 0.05), and increased MPS (p < 0.001). The results of this study indicate that 180 mg/L of GO and CIN promoted a more efficient rumen fermentation and increased the protein supply to the animal, respectively, although an apparent adaptive response of microbial populations to GO was observed.

Knowledge of Obesity's Health Related Outcomes among Hispanic Women living in Puerto Rico.

OBJECTIVE: The primary aim of this cross-sectional study was to assess, according to previous cancer diagnosis, the knowledge Puerto Rican women have on the link between obesity-endometrial, -breast, and colon cancer, and determine women's most common source for medical information. METHODS: In this cross-sectional study, eligible female patients (n=234) from the Gynecology and Gynecology-Oncology Clinics completed a self-administered survey from October 2014 to March 2016. Participants were evaluated on sociodemographic, body mass index, knowledge of the obesity-cancer link, and source of medical information. RESULTS: About 49% and 31% of women in the study were obese and overweight, respectively. Less than 52% of the women knew about the link between obesity-breast,-colon, and/or -endometrial cancers. Women with previous cancer diagnoses were more likely aware of the association between obesity-colon cancer than women without previous cancer diagnoses (58% vs. 44%, p0.05). Higher incomes showed increased odds for the knowledge obesity-cancer link among women with a cancer history, but the odds decreased for women without previous cancer diagnoses (p>0.05). Higher education showed a trend towards a better knowledge of the obesity-cancer association. The most common sources of information were the primary doctor (80%) and the internet (54%). CONCLUSION: Counseling about preventable cancer risk factors through primary care to all women must be encouraged especially in young healthy females. Further studies should address qualitative aspects of the odds differences in the obesity-cancer link knowledge observed between income categories for women with/without previous cancer diagnoses.

Systematic integration of m6A regulators and autophagy-related genes in combination with long non-coding RNAs predicts survival in glioblastoma multiforme.

Glioblastoma multiforme (GBM) is probably the only tumor in which a unique epigenetic alteration, namely methylation of the MGMT gene, possesses direct clinical relevance. Now with the emergence of aberrant N6 methyladenosine (m6A) modifications (the most common epigenetic modification of mRNA, closely linked to the autophagy process) in cancer, the epi-transcriptomic landscape of GBM pathobiology has been expanded. Considering this, herein, we systematically analyzed m6A regulators, assessed their correlation with autophagy-related genes (ATG), and established a long non-coding RNAs (lncRNA)-dependent prognostic signature (m6A-autophagy-lncRNAs) for GBM. Our analysis identified a novel signature of five long non-coding RNAs (lncRNAs: ITGA6-AS1, AC124248.1, NFYC-AS1, AC025171.1, and AC005229.3) associated with survival of GBM patients, and four among them clearly showed cancer-associated potential. We further validated and confirmed the altered expression of two lncRNAs (AC124248.1, AC005229.3) in GBM associated clinical samples using RT-PCR. Concerning the prognostic ability, the obtained signature determined high-/low-risk groups in GBM patients and showed sensitivity to anticancer drugs. Collectively, the m6A-autophagy-lncRNAs signature presented in the study is clinically relevant and is the first attempt to systematically predict the potential interaction between the three key determinants (m6A, autophagy, lncRNA) in cancer, particularly in GBM.

Jumping vs. running: Effects of exercise modality on aerobic capacity and neuromuscular performance after a six-week high-intensity interval training.

PURPOSE: High-intensity interval training (HIIT) has proven to be effective in improving endurance capacity and muscle endurance. However, its potential to improve other aspects of physical performance such as strength and power has not been well explored, and most research studies used only running and cycling as exercise modalities. Here, we compared the effects of jumping versus running as exercise modalities during a 6-week HIIT. METHODS: 46 participants (24±3 years, 171±9 cm, 68±13 kg, 22 women) were randomly allocated to one of three groups: countermovement jump training, running training, or control. The two training groups underwent a 6-week HIIT with 3 training sessions per week. Both training protocols had identical training frequency, number of series and work/rest durations (on average 7 series of 25s, with a rest of 25s between series). Before and after the training period, aerobic capacity and neuromuscular performance were assessed. RESULTS: Analyses of variance revealed a significant group*time interaction effect for maximal aerobic capacity (p = 0.004), and post hoc analyses showed a significant increase in the running group (p < .001, +7.6%). Analyses of the maximal voluntary contraction revealed only a significant increase in the jumping group (plantar flexion +12.8%, knee extension +8.2%). No interaction effects were found for maximal power or jump height. CONCLUSION: Despite identical programming, the choice of exercise mode profoundly affected the training adaptations: the running group significantly increased aerobic capacity, and the jump group significantly increased leg strength. These results underline the importance of exercise modality in physical performance adaptations.

Small RNAs and their protein partners in animal meiosis.

Meiosis is characterized by highly regulated transitions in gene expression that require diverse mechanisms of gene regulation. For example, in male mammals, transcription undergoes a global shut-down in early prophase I of meiosis, followed by increasing transcriptional activity into pachynema. Later, as spermiogenesis proceeds, the histones bound to DNA are replaced with transition proteins, which are themselves replaced with protamines, resulting in a highly condensed nucleus with repressed transcriptional activity. In addition, two specialized gene silencing events take place during prophase I: meiotic silencing of unsynapsed chromatin (MSUC), and the sex chromatin specific mechanism, meiotic sex chromosome inactivation (MSCI). Notably, conserved roles for the RNA binding protein (RBP) machinery that functions with small non-coding RNAs have been described as participating in these meiosis-specific mechanisms, suggesting that RNA-mediated gene regulation is critical for fertility in many species. Here, we review roles of small RNAs and their associated RBPs in meiosis-related processes such as centromere function, silencing of unpaired chromatin and meiotic recombination. We will discuss the emerging evidence of non-canonical functions of these components in meiosis.

ZBTB18 inhibits SREBP-dependent lipid synthesis by halting CTBPs and LSD1 activity in glioblastoma.

Enhanced fatty acid synthesis is a hallmark of tumors, including glioblastoma. SREBF1/2 regulate the expression of enzymes involved in fatty acid and cholesterol synthesis. Yet, little is known about the precise mechanism regulating SREBP gene expression in glioblastoma. Here, we show that a novel interaction between the co-activator/co-repressor CTBP and the tumor suppressor ZBTB18 regulates the expression of SREBP genes. In line with our findings, metabolic assays and glucose tracing analysis confirm the reduction in several phospholipid species upon ZBTB18 expression. Our study identifies CTBP1/2 and LSD1 as co-activators of SREBP genes and indicates that the functional activity of the CTBP-LSD1 complex is altered by ZBTB18. ZBTB18 binding to the SREBP gene promoters is associated with reduced LSD1 demethylase activity of H3K4me2 and H3K9me2 marks. Concomitantly, the interaction between LSD1, CTBP, and ZNF217 is increased, suggesting that ZBTB18 promotes LSD1 scaffolding function. Our results outline a new epigenetic mechanism enrolled by ZBTB18 and its co-factors to regulate fatty acid synthesis that could be targeted to treat glioblastoma patients.

Potential of Agroindustrial By-Products to Modulate Ruminal Fermentation and Reduce Methane Production: In Vitro Studies.

The effects of including wine lees (WL), exhausted olive cake (EOC) and a 1:1 mixture of EOC and tomato pomace (EOCTP) in diets for fattening ruminants on in vitro fermentation parameters and CH4 production were analysed. Ten diets were studied, containing either none of the tested by-products (control), or 6.0, 12.0 or 18.0% of WL, EOC and ECOTP formulated to have similar protein and fiber content. Diets were incubated in vitro with sheep ruminal fluid to measure gas production kinetics and fermentation parameters. Increasing the level of WL, EOC and EOCTP decreased linearly (p ≤ 0.009) the potential gas production, but other gas production parameters were unaffected (p > 0.05), excepting that EOCTP increased the gas production rate. No differences (p ≥ 0.0.05) among diets were observed in total volatile fatty acid (VFA) production at 24 h of incubation for EOC and EOCTP, but NH3-N concentration decreased (p ≤ 0.003). In contrast, WL at 12.0 and 18.0% decreased (p < 0.05) total VFA production and increased the acetate/propionate ratio (p < 0.05). None of the by-products had an effect on CH4 production (p ≥ 0.0.05). Results indicate that EOC and EOCTP could be included up to 18.0% in fattening diets, but lower levels of WL are recommended.

Test-Retest Reliability and Sensitivity of Common Strength and Power Tests over a Period of 9 Weeks.

This study evaluated the reliability and sensitivity of a set of different common strength and power tests in a healthy adult population in a span of 9 weeks. Seventeen subjects (24.2 ± 2.2 years, 1.75 ± 0.10 m, 68.6 ± 14.2 kg, seven women) participated in the study. We tested countermovement jumps, reactive hops, and the maximal voluntary contraction (MVC) of handgrip and isometric knee extension. The tests were conducted in three separate sessions across a nine-week period, with one week between the first two sessions and eight weeks between the second and the third. Reliability and sensitivity statistics for each test were calculated for both the average of three trials and the best result during each session. The MVC of isometric knee extension and handgrip, as well as the countermovement jump test, demonstrated very high reliability and sensitivity over the nine-week period. The peak force of the reactive hops demonstrated high reliability but high sensitivity only for the average but not for the best result. The average contact time of reactive hops was neither a sensitive nor reliable measurement. In conclusion, isometric maximal knee extension and handgrip tests, as well as countermovement jumps and peak force of reactive hops, can be used as reliable and sensitive measurements of isometric and reactive strength and power over time periods of up to eight weeks. We recommend the use of the average results of three trials instead of the best performance value for longitudinal studies, as this procedure produces more consistent results and a lower measurement error.

Calpain-mediated cleavage generates a ZBTB18 N-terminal product that regulates HIF1A signaling and glioblastoma metabolism.

Proteolytic cleavage is an important post-translational mechanism to increase protein variability and functionality. In cancer, this process can be deregulated to shut off tumor-suppressive functions. Here, we report that in glioblastoma (GBM), the tumor suppressor ZBTB18 is targeted for protein cleavage by the intracellular protease calpain. The N-terminal (Nte) ZBTB18 cleaved fragment localizes to the cytoplasm and thus, is unable to exert the gene expression repressive function of the uncleaved protein. Mass spectrometry (MS) analysis indicates that the Nte ZBTB18 short form (SF) interacts with C-terminal (Cte) binding proteins 1 and 2 (CTBP1/2), which appear to be involved in HIF1A signaling activation. In fact, we show that the new ZBTB18 product activates HIF1A-regulated genes, which in turn lead to increased lipid uptake, lipid droplets (LD) accumulation, and enhanced metabolic activity. We propose that calpain-mediated ZBTB18 cleavage represents a new mechanism to counteract ZBTB18 tumor suppression and increase tumor-promoting functions in GBM cells.

T-cell dysfunction in the glioblastoma microenvironment is mediated by myeloid cells releasing interleukin-10.

Despite recent advances in cancer immunotherapy, certain tumor types, such as Glioblastomas, are highly resistant due to their tumor microenvironment disabling the anti-tumor immune response. Here we show, by applying an in-silico multidimensional model integrating spatially resolved and single-cell gene expression data of 45,615 immune cells from 12 tumor samples, that a subset of Interleukin-10-releasing HMOX1+ myeloid cells, spatially localizing to mesenchymal-like tumor regions, drive T-cell exhaustion and thus contribute to the immunosuppressive tumor microenvironment. These findings are validated using a human ex-vivo neocortical glioblastoma model inoculated with patient derived peripheral T-cells to simulate the immune compartment. This model recapitulates the dysfunctional transformation of tumor infiltrating T-cells. Inhibition of the JAK/STAT pathway rescues T-cell functionality both in our model and in-vivo, providing further evidence of IL-10 release being an important driving force of tumor immune escape. Our results thus show that integrative modelling of single cell and spatial transcriptomics data is a valuable tool to interrogate the tumor immune microenvironment and might contribute to the development of successful immunotherapies.

Expression of Chlamydomonas reinhardtii chloroplast diacylglycerol acyltransferase 3 is induced by light in concert with triacylglycerol accumulation.

Considerable progress has been made towards the understanding of triacylglycerol (TAG) accumulation in algae. One key aspect is finding conditions that trigger TAG production without reducing cell division. Previously, we identified a soluble diacylglycerol acyltransferase (DGAT), related to plant DGAT3, with heterologous DGAT activity. In this work, we demonstrate that Chlamydomonas reinhardtii DGAT3 localizes to the chloroplast and that its expression is induced by light, in correspondence with TAG accumulation. Dgat3 mRNAs and TAGs increase in both wild-type and starch-deficient cells grown with acetate upon transferring them from dark or low light to higher light levels, albeit affected by the particularities of each strain. The response of dgat3 mRNAs and TAGs to light depends on the pre-existing levels of TAGs, suggesting the existence of a negative regulatory loop in the synthesis pathway, although an effect of TAG turnover cannot be ruled out. Altogether, these results hint towards a possible role of DGAT3 in light-dependent TAG accumulation in C. reinhardtii.

Mitigation of ruminal methane production with enhancing the fermentation by supplementation of different tropical forage legumes.

The aim of this research was to evaluate the influence of forage species adapted to the tropical region of Ecuador on gas production, enteric methane, digestion, and ruminal fermentation. The tree forage evaluated were C. arborea, E. fusca, B. forficata, E. poeppigiana, C. argentea, G. sepium, C. tora, and F. macrophylla. Ruminal fluid of four adult sheep fistulated with permanent cannulas in the rumen was used in the in vitro gas production technique. The in vitro gas production parameters were lower (P < 0.05) in the C. arborea (A = 41.68 mL gas/g DM, c = 0.044%/h and Lag = 1.654 h) and the average gas production rate for B. forficata was 1.017 mL/h (P < 0.05). C. arborea presented higher (P = 0.0001) effective degradation and real DM digestibility (40.461 g/kg and 82.51 mg/g, respectively). With respect to VFA, the highest (P < 0.05) proportion of acetic, propionic, and butyric was observed in C. arborea, G. sepium, and E. poeppigiana (72.52, 23.09, and 7.44 mol/100 mol, respectively) and the lowest (P = 0.0001) ratio: acetic/propionic was observed in G. sepium (2.92 mol/100 mol). The content of NH3-N (mg/L) showed no difference. The lowest (P = 0.0001) methane production was observed in C. arborea (1.23 mL CH4/g DM). The use of forage species of tropical climate rich in secondary metabolites in ruminant diets has the capacity to reduce the gas production and enteric methane; however, this is at the expense of the reduction of the fermentation of organic matter in the rumen.

NF1 regulates mesenchymal glioblastoma plasticity and aggressiveness through the AP-1 transcription factor FOSL1.

The molecular basis underlying glioblastoma (GBM) heterogeneity and plasticity is not fully understood. Using transcriptomic data of human patient-derived brain tumor stem cell lines (BTSCs), classified based on GBM-intrinsic signatures, we identify the AP-1 transcription factor FOSL1 as a key regulator of the mesenchymal (MES) subtype. We provide a mechanistic basis to the role of the neurofibromatosis type 1 gene (NF1), a negative regulator of the RAS/MAPK pathway, in GBM mesenchymal transformation through the modulation of FOSL1 expression. Depletion of FOSL1 in NF1-mutant human BTSCs and Kras-mutant mouse neural stem cells results in loss of the mesenchymal gene signature and reduction in stem cell properties and in vivo tumorigenic potential. Our data demonstrate that FOSL1 controls GBM plasticity and aggressiveness in response to NF1 alterations.

Desmosterol Incorporation Into Ram Sperm Membrane Before Cryopreservation Improves in vitro and in vivo Fertility.

Pregnancy rates in ewes are markedly low after cervical insemination with frozen-thawed sperm. Sensitivity of ram sperm to freeze-thawing is related to the lipid composition of the membrane, particularly to its low sterol content. Recently, we proved that sterol content of ram sperm can be increased by treatment with methyl-ß-cyclodextrin-sterol complexes and we provided mechanistic based evidence on the differential behavior of cholesterol and desmosterol in the ram sperm membrane. In the present study, we evaluated the role of increasing cholesterol and desmosterol content of ram sperm before cryopreservation, on the extent and distribution of sterols, cryocapacitation status, acrosome integrity, DNA damage associated with apoptosis and fertility competence in vitro and in vivo of post-thawed sperm. After freeze-thawing, similar levels of sterol content were evidenced in control sperm cells and in those pre-incubated with either cholesterol or desmosterol. Still, moderately higher levels of sterols were registered in treated sperm compared to the control, indicating no physiological excess of sterols after thawing or sterol losses that exceed the control. Live cell imaging of fluorescent cholesterol evidenced the presence of sperm sub-populations differentially affected by freeze-thawing. Similar unimodal frequency profiles were observed between sterol-enriched groups, while the control exhibited a sub-population of sperm compatible with low sterol content. Tyrosine phosphorylation was significantly lower when ram sperm incorporated cholesterol compared to the control. No difference in this capacitation parameter was found between the latter and desmosterol-enriched sperm. The percentage of sperm with damaged acrosomes post-thawing, assessed by a fluorescent lectin, was reduced in sperm that incorporated sterols before freezing, irrespective of the sterol class. These results suggest that sterols exert a stabilizing effect on the acrosome. No differences were found in levels of apoptotic DNA fragmentation among experimental groups. As to fertility trials, desmosterol-enriched sperm gave rise to higher rates of in vitro activated oocytes by heterologous fertilization and to significantly lower pregnancy loss in vivo. Our research provides new insights on sterol incorporation into ram sperm prior to cryopreservation, in particular on the additional benefit of incorporating desmosterol as a strategy to improve fertility outcome.

Daily 30-min exposure to artificial gravity during 60 days of bed rest does not maintain aerobic exercise capacity but mitigates some deteriorations of muscle function: results from the AGBRESA RCT.

PURPOSE: Spaceflight impairs physical capacity. Here we assessed the protective effect of artificial gravity (AG) on aerobic exercise capacity and muscle function during bed rest, a spaceflight analogue. METHODS: 24 participants (33 ± 9 years, 175 ± 9 cm, 74 ± 10 kg, 8 women) were randomly allocated to one of three groups: continuous AG (cAG), intermittent AG (iAG) or control (CTRL). All participants were subjected to 60 days of six-degree head-down tilt bed rest, and subjects of the intervention groups completed 30 min of centrifugation per day: cAG continuously and iAG for 6 × 5 min, with an acceleration of 1g at the center of mass. Physical capacity was assessed before and after bed rest via maximal voluntary contractions, cycling spiroergometry, and countermovement jumps. RESULTS: AG had no significant effect on aerobic exercise capacity, flexor muscle function and isometric knee extension strength or rate of force development (RFD). However, AG mitigated the effects of bed rest on jumping power (group * time interaction of the rmANOVA p < 0.001; iAG - 25%, cAG - 26%, CTRL - 33%), plantar flexion strength (group * time p = 0.003; iAG - 35%, cAG - 31%, CTRL - 48%) and plantar flexion RFD (group * time p = 0.020; iAG - 28%, cAG - 12%, CTRL - 40%). Women showed more pronounced losses than men in jumping power (p < 0.001) and knee extension strength (p = 0.010). CONCLUSION: The AG protocols were not suitable to maintain aerobic exercise capacity, probably due to the very low cardiorespiratory demand of this intervention. However, they mitigated some losses in muscle function, potentially due to the low-intensity muscle contractions during centrifugation used to avoid presyncope.

Utilization of Avocado and Mango Fruit Wastes in Multi-Nutrient Blocks for Goats Feeding: In Vitro Evaluation.

This study was conducted to investigate the nutritive value of avocado and mango fruit wastes, and to assess the possibility of preserving the wastes into multi-nutrient blocks (MB). Both peels and a pulp:peels (PP) mixture of each fruit were analyzed for chemical composition and in vitro fermentation with goats' ruminal fluid. Wastes had low-dry matter (DM) content (<250 g/kg), with those from mango having high non-structural carbohydrates content (>800 g/kg DM) and those from avocado high fat levels (>580 g/kg DM). Mango wastes were fermented at a greater extent and faster rate than avocado ones. The PP mixture of each fruit was included into multi-nutrient blocks (MB) formulated to have similar chemical composition. There were only subtle differences in the fermentation of MB including wastes from either avocado or mango, but fermentation of avocado-MB resulted in significantly (p ≤ 0.032) greater acetate and lower propionate proportions than mango-MB. Including the PP mixture in the formulation of MB for goats feeding is a feasible option to reduce the environmental impact of avocado and mango fruit wastes, but studies on the acceptance of the MB by goats and their stability over long-time storage periods are needed.

Replacing Forage by Crude Olive Cake in a Dairy Sheep Diet: Effects on Ruminal Fermentation and Microbial Populations in Rusitec Fermenters.

Olive oil extraction generates large amounts of a highly pollutant by-product called olive cake (OC), and its use in ruminant feeding could be an alternative. This study was designed to evaluate the effects of partially replacing forage by crude OC (COC) in a mixed dairy diet on rumen fermentation and microbial populations in Rusitec fermenters. The COC replaced 33% of the forage (66% maize silage and 33% barley straw) and was included at 16.6% of the total diet. Four fermenters were used in a cross-over design with two 13-day incubation periods. Experimental diets had a 50:50 forage-to-concentrate ratio and were formulated to contain the same protein (16.0%) and neutral detergent fiber (32.5%) levels. Compared with control fermenters, those fed the COC diet showed greater (p ≤ 0.02) pH (6.07 vs. 6.22), diet disappearance (0.709 vs. 0.748), and butyrate proportions (18.0 vs. 19.4), but there were no differences in volatile fatty acids and ammonia production. Microbial growth, bacterial diversity, protozoal abundance, and relative abundance of fungi and archaea were unaffected by diet, although the solid phase of COC-fed fermenters showed greater (p = 0.01) bacterial abundance than control ones. Results indicate that COC could replace 33% of the forage in a mixed dairy diet.

In Vitro and In Situ Evaluation of Broccoli Wastes as Potential Feed for Ruminants.

The potential of broccoli wastes (florets and stems) as ruminant feed was analyzed using in vitro and in situ techniques. Both stems and florets had high moisture content (90.6 and 86.1%, respectively), but the stems contained (% dry matter) lower levels (p < 0.05) of crude protein (CP; 23.2 vs. 30.8%) and ether extract (2.91 vs. 6.15%) and tended to have greater sugars content (p = 0.071; 33.4 vs. 19.6%) than florets. Stems had greater in vitro dry matter rumen degradability (45.3%; 24 h incubation) and lower in vitro CP intestinal digestibility (82.7%) compared with florets (42.2 and 90.1%, respectively). Rumen degradability of protein was high (<85%) for both fractions. In a second experiment, diets including different proportions of broccoli were formulated and fermented in vitro. The replacement of 24% of conventional feeds (wheat, soybean meal and wheat bran) in a concentrate by dried broccoli increased the amount of organic matter fermented in vitro and the NH3-N concentrations of a mixed diet including 40% of the concentrate. Including dried broccoli in the diet produced only small modifications in the volatile fatty acid profile and did not affect CH4 emission.