Supplementation of lactobacillus-fermented rapeseed meal in broiler diet reduces Campylobacter jejuni cecal colonization and limits the l-tryptophan and l-histidine biosynthesis pathways.

BACKGROUND: Campylobacter jejuni (C. jejuni), a widely distributed global foodborne pathogen, primarily linked with contaminated chicken meat, poses a significant health risk. Reducing the abundance of this pathogen in poultry meat is challenging but essential. This study assessed the impact of Lactobacillus-fermented rapeseed meal (LFRM) on broilers exposed to C. jejuni-contaminated litter, evaluating growth performance, Campylobacter levels, and metagenomic profile. RESULTS: By day 35, the litter contamination successfully colonized broilers with Campylobacter spp., particularly C. jejuni. In the grower phase, LFRM improved (P < 0.05) body weight and daily weight gain, resulting in a 9.2% better feed conversion ratio during the pre-challenge period (the period before artificial infection; days 13-20). The LFRM also reduced the C. jejuni concentration in the ceca (P < 0.05), without altering alpha and beta diversity. However, metagenomic data analysis revealed LFRM targeted a reduction in the abundance of C. jejuni biosynthetic pathways of l-tryptophan and l-histidine and gene families associated with transcription and virulence factors while also possibly leading to selected stress-induced resistance mechanisms. CONCLUSION: The study demonstrated that LFRM inclusion improved growth and decreased cecal Campylobacter spp. concentration and the relative abundance of pivotal C. jejuni genes. Performance benefits likely resulted from LFRM metabolites. At the molecular level, LFRM may have reduced C. jejuni colonization, likely by decreasing the abundance of energy transduction and l-histidine and l-tryptophan biosynthesis genes otherwise required for bacterial survival and increased virulence. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effect of dietary protein source and Saccharina latissima on nutritional and safety characteristics of milk.

BACKGROUND: Wheat distillers' grains (WDG) and seaweeds are recommended as alternative protein sources and enteric methane mitigators in dairy cow diets, respectively, but little is known about their impact on milk quality and safety. In the present study, 16 cows in four 4 × 4 Latin squares were fed isonitrogenous diets (50:50 forage:concentrate ratio), with rapeseed meal (RSM)-based or WDG-based concentrate (230 and 205 g kg-1 dry matter) and supplemented with or without Saccharina latissima. RESULTS: Replacement of RSM with WDG enhanced milk nutritional profile by decreasing milk atherogenicity (P = 0.002) and thrombogenicity (P = 0.019) indices and the concentrations of the nutritionally undesirable saturated fatty acids - specifically, lauric (P = 0.045), myristic (P = 0.022) and palmitic (P = 0.007) acids. It also increased milk concentrations of the nutritionally beneficial vaccenic (P < 0.001), oleic (P = 0.030), linoleic (P < 0.001), rumenic (P < 0.001) and α-linolenic (P = 0.012) acids, and total monounsaturated (P = 0.044), polyunsaturated (P < 0.001) and n-6 (P < 0.001) fatty acids. Feeding Saccharina latissima at 35.7 g per cow per day did not affect the nutritionally relevant milk fatty acids or pose any risk on milk safety, as bromoform concentrations in milk were negligible and unaffected by the dietary treatments. However, it slightly reduced milk concentrations of pantothenate. CONCLUSION: Feeding WDG to dairy cows improved milk fatty acid profiles, by increasing the concentrations of nutritionally beneficial fatty acids and reducing the concentration of nutritionally undesirable saturated fatty acids, while feeding seaweed slightly reduced pantothenate concentrations. However, when considering the current average milk intakes in the population, the milk compositional differences between treatments in this study appear relatively small to have an effect on human health. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Novel sfericase protease improves amino acid digestibility of soybean meal and rapeseed meal in broiler chickens.

1. Two experiments were conducted to explore the effects of an exogenous sfericase protease on the apparent ileal nutrient digestibility of soybean meal (SBM) and rapeseed meal (RSM) in broiler chickens.2. In each experiment, a total of 256 sixteen-day-old male Cobb 500 broilers were fed one of four semi-purified experimental diets, comprising two different batches (A and B) of samples for either SBM (Exp. 1) or RSM (Exp. 2) without or with an exogenous sfericase (0 or 30,000 NFP/kg). Each experimental diet was fed to eight replicate pens of broiler chickens from 16 to 21 d of age (eight birds per cage), and ileal digesta were collected for measuring the digestibility coefficients.3. In Exp. 1, the amino acid digestibility was greater (P < 0.05) in SBM B compared with SBM A for Arg and Val, and a similar trend (P < 0.1) was observed for Tyr, Leu and Thr. Exogenous sfericase increased (P < 0.10) digestibility of most of amino acids except Gly, His and Trp. There was an interaction between SBM source and sfericase, whereby digestibility of P, N and Asp was increased by sfericase for SBM B but not for SBM A. In Exp. 2, there was no interaction (P > 0.05) between RSM source and sfericase for ileal nutrient digestibility. Digestibility was greater in RSM A compared to RSM B for all non-essential AA and most essential AA (except for Trp), while the reverse was noted for Ca and P (P < 0.05). Exogenous sfericase increased (P < 0.1) digestibility for all amino acids except Cys and Met.4. In conclusion, the current studies showed that both SBM and RSM batches influenced amino acid digestibility. Sfericase protease supplementation increased amino acid digestibility for both SBM and RSM. The digestibility effects were greater in the SBM batch with low digestibility for N and Asp which was in line with an increase in P digestibility.

Replacing soybean meal with fermented rapeseed meal in diets: potential effects on growth performance, antioxidant capacity, and liver and intestinal health of juvenile tilapia (Oreochromis niloticus).

This study aims to evaluate the effects of substituting soybean meal with fermented rapeseed meal (FRM) on growth, antioxidant capacity, and liver and intestinal health of the genetically improved farmed tilapia (GIFT, Oreochromis niloticus). A total of 450 tilapia (7.22 ± 0.15 g) were fed with five experimental diets, including a basal diet containing 40% soybean meal (CP0), which was subsequently replaced by 25% (CP25), 50% (CP50), 75% (CP75), and 100% (CP100) FRM in a recirculated aquiculture system for 9 weeks (30 fish per tank in triplicates). The results showed that the weight gain, specific growth rate, feed intake, feed efficiency, hepatosomatic index, and viscerosomatic index of fish in both CP75 and CP100 groups were significantly lower than those in CP0 group (P < 0.05). The fish in CP100 group had the lower content of muscle crude protein while the higher level of muscle crude lipid (P < 0.05). Activities of serum aspartate aminotransferase, alanine aminotransferase along with total triglyceride in CP100 group were significantly higher than those in CP0 group (P < 0.05). There were no significant differences in the contents of liver protease, amylase, and lipase among five groups (P > 0.05). The activities of liver total antioxidant capacity and superoxide dismutase exhibited the increased tendency with the increase of FRM replacement levels from 25 to 50% (P < 0.05), while then significantly decreased from 75 to 100% (P < 0.05). Histological morphology indicated that the fish in between CP75 and CP100 groups had poor liver and intestine health. Intestinal microbial diversity analysis showed that the relative abundance of Cetobacterium and Alcaligenaceae in both CP75 and CP100 groups were lower than that in other three groups. In conclusion, the maximum replacement level of soybean meal with FRM in the diet was determined to be 50% without compromising the growth performance, antioxidant status, and liver and intestinal health of tilapia under the current experimental conditions. The observed decrease in food intake and subsequent retarded growth performance in the CP75 and CP100 groups can be attributed directly to a reduction in feed palatability caused by FRM.

Processed fava bean as a substitute for rapeseed meal with or without rumen-protected methionine supplement in grass silage-based dairy cow diets.

Fava bean offers a sustainable home-grown protein source for dairy cows, but fava bean protein is extensively degraded in the rumen and has low Met concentration. We studied the effects of protein supplementation and source on milk production, rumen fermentation, N use, and mammary AA utilization. The treatments were unsupplemented control diet, and isonitrogenously given rapeseed meal (RSM), processed (dehulled, flaked, and heated) fava bean without (TFB) or with rumen-protected (RP) Met (TFB+). All diets consisted of 50% grass silage and 50% cereal-based concentrate including studied protein supplement. The control diet had 15% of crude protein and protein-supplemented diets 18%. Rumen-protected Met in TFB+ corresponded to 15 g/d of Met absorbed in the small intestine. Experimental design was a replicated 4 × 4 Latin square with 3-wk periods. The experiment was conducted using 12 multiparous mid-lactation Nordic Red cows, of which 4 were rumen cannulated. Protein supplementation increased dry matter intake (DMI), and milk (31.9 vs. 30.7 kg/d) and milk component yields. Substituting RSM with TFB or TFB+ decreased DMI and AA intake but increased starch intake. There were no differences in milk yield or composition between RSM diet and TFB diets. Rumen-protected Met did not affect DMI, or milk or milk component yields but increased milk protein concentration in comparison to TFB. There were no differences in rumen fermentation except for increased ammonium-N concentration with the protein-supplemented diets. Nitrogen-use efficiency for milk production was lower for the supplemented diets versus control diet but tended to be greater for TFB and TFB+ versus RSM. Protein supplementation increased plasma essential AA concentration but there were no differences between TFB diets and RSM. Rumen-protected Met clearly increased plasma Met concentration (30.8 vs. 18.2 µmol/L) but did not affect other AA. Absence of differences between RSM and TFB in milk production together with limited effects of RP Met suggest that TFB is a potential alternative protein source for dairy cattle.

Biodegradation of nitriles derived from glucosinolates in rapeseed meal by BnNIT2: a nitrilase from Brassica napus with wide substrate specificity.

Nitriles derived from glucosinolates (GSLs) in rapeseed meal (RSM) can cause lesions on animal liver and kidneys. Nitrilase converts nitriles to carboxylic acids and NH3, eliminating their toxicity. Here we describe a nitrilase, BnNIT2, from Brassica napus (optimal temperature, 45 °C; pH, 7.0) that is stable at 40 °C and has a wide substrate specificity. Recombinant BnNIT2 converted the three main nitriles from GSLs (3-hydroxy-4-pentenenitrile, 3-butenenitrile, and 4-pentenenitrile), with the highest specific activity (58.6 U/mg) for 4-pentenenitrile. We used mutagenesis to improve the thermostability of BnNIT2; the resulting mutant BnNIT2-H90M had an ~ 14.5% increase in residual activity at 50 °C for 1 h. To verify the functionality of BnNIT2, GSLs were extracted from RSM and converted into nitriles at pH 5.0 in the presence of Fe2+. Then, BnNIT2 was used to degrade the nitriles from GSLs; ultimately, ~ 80% of nitriles were removed. Thus BnNIT2 is a potential enzyme for detoxification of RSM. KEY POINTS: • Functional identification of the plant nitrilase BnNIT2. • Identified a mutant, H90M, with improved thermostability. • BnNIT2 was capable of degrading nitriles from transformed GSLs.

Yeast-Fermented Rapeseed Meal Extract Is Able to Reduce Inflammation and Oxidative Stress Caused by Escherichia coli Lipopolysaccharides and to Replace ZnO in Caco-2/HTX29 Co-Culture Cells.

(1) The present study tested in vitro the capacity of a fermented rapeseed meal extract to reduce medicinal ZnO, which will be banned at the EU level from 2023 onwards because of its potential to cause environmental pollution and the development of Zn resistance in gut bacteria. Rapeseed meal could be an important ZnO substitute as it has antioxidant/radical scavenging properties due to its content of bioactive compounds (e.g., polyphenols). (2) Protein array and flow cytometry were used to detect apoptosis, oxidative stress production, and inflammatory and signaling-related molecules in Caco-2 and goblet HT29-MTX co-culture cells challenged with Escherichia coli lipopolysaccharides and treated with ZnO and FRSM. (3) LPS induced cell death (21.1% vs. 12.7% in control, p < 0.005); apoptosis (16.6%); ROS production; and overexpression of biomarkers related to inflammation (63.15% cytokines and 66.67% chemokines), oxidative stress, and signaling proteins when compared to untreated cells. ZnO was effective in counteracting the effect of LPS, and 73.68% cytokines and 91.67% of chemokines were recovered. FRSM was better at restoring normal protein expression for 78.94% of cytokines, 91.67% of chemokines, and 61.11% of signaling molecules. FRSM was able to mitigate negative effects of LPS and might be an alternative to ZnO in pig diets.

In situ ruminal disappearance of crude protein and phytate from differently processed rapeseed meals in dairy cows.

BACKGROUND: The influence of different processing conditions of rapeseed meal on ruminal degradation of crude protein and phytate in dairy cows was investigated. Following oil extraction from the rapeseed, five residence times in the desolventizer/toaster were chosen to remove the solvent from the meal. Rapeseed cake and rapeseed meals were incubated in situ in the rumen of three fistulated dairy cows to determine ruminal degradation parameters. RESULTS: With increasing residence time in the desolventizer/toaster the ruminal degradation of crude protein decreased significantly for every treatment step. Ruminal phytate degradation and crude protein degradation were affected almost identically. CONCLUSION: The processing conditions of rapeseed meal have a major impact on the ruminal degradation of crude protein and phytate, indicating a potential conflict of interest regarding the production process. Large amounts of undegradable rumen protein are often intended for high-yielding dairy cows whereas a high level of ruminal degradation is preferred for phytate to increase absorption of phosphorus in the small intestine. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effect of rapeseed meal degraded by enzymolysis and fermentation on the growth performance, nutrient digestibility and health status of broilers.

The purpose of this study is to investigate the nutritional changes of degraded rapeseed meal and its effects on growth performance, nutrient digestibility and health status of broilers. Raw rapeseed meal (CON), degraded by enzymolysis (protease, ERM), fermentation (Bacillus subtilis, FRM) or both (DRM) were included in diets at 25% and fed to 480 yellow-feathered broilers at 22-63 d of age. Results showed that rapeseed peptide contents (≤1 kDa) were increased (p < 0.05) from 4.13% (CON) to 35.5% (ERM), 24.1% (FRM) and 50.4% (DRM); glucosinolate and erucic acid in DRM were decreased (p < 0.05) by 71.6% and 86.2%, respectively, compared to CON. There were increases (p ≤ 0.029) in feed intake, body weight gain, feed efficiency and precaecal digestibility of dry matter, crude protein, methionine, isoleucine, leucine, lysine, cysteine, phenylalanine, tyrosine, threonine, tryptophan and valine in the three degraded diets. Also, serum immunoglobulin (Ig) A, IgG, glutathione peroxidase, superoxide dismutase and catalase were raised (p ≤ 0.034) in the degraded diets. Additionally, DRM showed more pronounced effects (p < 0.05) on variables related to growth, digestibility and health than ERM and FRM. It is concluded that rapeseed meal degraded by both enzymolysis and fermentation can increase its nutritional values and application in broilers.

The effect of fermented and raw rapeseed meal on the growth performance, immune status and intestinal morphology of broiler chickens.

We evaluated the effect of fermented and raw rapeseed meal (FRCM and RRSM) on the growth performance, carcass traits, serum biochemical indexes, immune status and intestinal morphology of broilers. A total of 420-day-old Arbor Acre male broilers were randomly assigned to a 1 + 2 × 3 factorial arrangement with one basal diet group, two rapeseed meal (RSM) varieties (FRCM and RRSM) and three addition levels (5%, 10% and 15%) for a duration of 42 days. FRSM significantly increased the ADG and ADFI of broilers during the 22-42 days and 1-42 days (p < 0.05) growth periods compared with RRSM groups. No significant difference was observed in ADG and ADFI between broilers fed FRSM in different dietary levels and control diets (p > 0.05), but broilers fed diet with 15% RRSM showed significantly lower ADG, ADFI and spleen index (p < 0.05). Both FRSM and RRSM in different dietary levels affected the anti-oxidation function of broilers, including a significant increase in the serum contents of superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) (p < 0.05), and a significant decrease of malondialdehyde (MDA) (p < 0.05). Fermentation tends to increase the villus height of duodenum (p = 0.09), and significantly increased the villus height and V/C ratio of duodenum (p < 0.05) in birds. The duodenum villus height was the highest in the 5% FRSM group and lowest in birds of 5% RRSM group. A 10% level significantly increased the duodenal V/C ratio in both FRSM and RRSM birds. The results indicated that FRSM could be used to supplemented in broilers diet to improve the production performance and maintain good health. FRSM can be added to broilers diet at 10% without adverse effect on growth performance or immune function.

Ruminal and post-ruminal phytate degradation of diets containing rapeseed meal or soybean meal.

This study aimed to investigate ruminal and post-ruminal degradation of phytic acid (InsP6) in diets containing either rapeseed meal (RSM) or soybean meal (SBM). In Experiment 1, the effective degradability of crude protein (CPED) and InsP6 (InsP6ED) was evaluated by incubating RSM and SBM in situ in three rumen-fistulated lactating Jersey cows for 2, 4, 6, 8, 16, 24, 48 and 72 h, and calculating effective degradability at rumen passage rates of 2% and 5%/h. In Experiment 2, eight wethers were assigned for 8 weeks to two dietary treatments (Diet RSM and Diet SBM) containing 150 g of either meal and 100 g of maize silage per feeding time and had free access to hay and water. Titanium dioxide (TiO2) was added to the diets for the last 5 days of the study. The wethers were then stunned, exsanguinated and digesta from the reticulo-rumen, omasum, abomasum, jejunum, colon, and rectum were sampled. In Experiment 1, the InsP6ED of RSM (InsP6ED2: 83%; InsP6ED5: 64%) decreased almost identically to that of CPED with increasing passage rate (CPED2: 78%; CPED5: 63%) and was significantly lower than that of SBM (InsP6ED2: 93%; InsP6ED5: 85%). In Experiment 2, ruminal InsP6 disappearance was significantly higher in wethers fed Diet SBM (89%) than in those fed Diet RSM (76%). Total post-ruminal InsP6 degradation was 6% for Diet RSM and 4% for Diet SBM (p = 0.186). The total tract InsP6 disappearance was higher in Diet SBM (93%) than in Diet RSM (82%). Considering higher InsP6 contents in RSM, Diet RSM resulted in significantly higher amounts of ruminally (Diet RSM: 4.5 g/d; Diet SBM: 3.4 g/d) and total tract (Diet RSM: 4.9 g/d; Diet SBM: 3.5 g/d) degraded InsP6. InsP5 was quantified in most of the digesta samples after feeding Diet RSM but was not detectable in the majority of digesta samples for Diet SBM. Concentrations of myo-inositol (MI) tended to be higher (p = 0.060) in the blood plasma of wethers fed Diet RSM. The consistency between ruminal InsP6 disappearance in wethers and in situ calculated InsP6ED2, along with the very low extent of post-ruminal InsP6 degradation, suggests that at a low rumen passage rate, InsP6-P from the feed becoming available to ruminants is almost entirely from InsP6 degradation in the rumen.

Effects of Achyranthes japonica extract on the performance of finishing pigs fed diets containing palm kernel meal and rapeseed meal as a partial alternative to soybean meal.

A total of 120 finishing pigs with an average initial body weight of 49.72 ± 0.08 kg (mean ± SD) were used in a 10 weeks trial. Pigs were randomly allotted into one of four dietary treatments (6 replicate pen/treatment, 5 pigs/pen). The nutritional dietary treatments were corn, soy bean meal, palm- kernel meal, and rapeseed meal based basal diets supplemented with 0, 0.05, 0.10, and 0.20% of Achyranthes japonica extract (AJE). Dietary inclusion of AJE supplementation had trend to increase the body weight and average daily gain of pigs at week 10 and the overall experimental period, respectively. The graded level of AJE supplement had increase the total track digestibility dry matter (p = 0.067) only at week 5 while nitrogen and energy digestibility (p < 0.05) was linearly increased at both weeks 5 and 10. During week 10, pigs fed with an increased level of AJE supplementation had linearly increase (p < 0.05) fecal Lactobacillus counts. In addition, AJE supplementation in the diet of finishing pigs had linearly decreased (p > 0.05) NH3 emission of gas and trend to decrease total mercaptans during week 10. Dietary inclusion of AJE supplement resulted in a linear increase in the blood protein concentration level. Moreover, drip loss was linearly reduced on day 5 and day 7 (p > 0.05) post slaughter in finishing pigs fed with gradually increased levels of AJE supplementation. During weeks 5 and 10, pigs fed with graded levels of AJE supplementation had linearly increase (p < 0.05) the backfat thickness and lean meat percentage. Therefore we conclude that dietary inclusion of AJE with palm kernel meal and rapeseed meal could be benificial to enhance the growth performance, nutrient digestibility, fecal microbial, blood prolife, meat quality and reduced fecal gas emission in finishing pigs.

Identification and Capture of Phenolic Compounds from a Rapeseed Meal Protein Isolate Production Process By-Product by Macroporous Resin and Valorization Their Antioxidant Properties.

In this study, phenolic compounds from an aqueous protein by-product from rapeseed meal (RSM) were identified by HPLC-DAD and HPLC-ESI-MS, including sinapine, sinapic acid, sinapoyl glucose, and 1,2-di-sinapoyl gentibiose. The main phenolic compound in this by-product was sinapine. We also performed acid hydrolysis to convert sinapine, and sinapic acid derivatives present in the permeate, to sinapic acid. The adsorption of phenolic compounds was investigated using five macroporous resins, including XAD4, XAD7, XAD16, XAD1180, and HP20. Among them, XAD16 showed the highest total phenolic contents adsorption capacities. The adsorption behavior of phenolic compounds was described by pseudo-second-order and Langmuir models. Moreover, thermodynamics tests demonstrated that the adsorption process of phenolic compounds was exothermic and spontaneous. The highest desorption ratio was obtained with 30% (v/v) and 70% (v/v) ethanol for sinapine and sinapic acid, respectively, with a desorption ratio of 63.19 ± 0.03% and 94.68 ± 0.013%. DPPH and ABTS tests revealed that the antioxidant activity of the hydrolyzed fraction was higher than the non-hydrolyzed fraction and higher than the one of vitamin C. Antioxidant tests demonstrated that these phenolic compounds could be used as natural antioxidants, which can be applied in the food industry.

Valorization Potentials of Rapeseed Meal in a Biorefinery Perspective: Focus on Nutritional and Bioactive Components.

Rapeseed meal (RSM), a by-product of oilseed extraction connected to the agri-food and biofuel sectors, is currently used as animal feed and for other low-value purposes. With a biorefinery approach, RSM could be valorized as a source of bio-based molecules for high-value applications. This study provides a chemical characterization of RSM in the perspective of its valorization. A qualitative study of main functional groups by fourier transform infrared (FTIR) spectroscopy was integrated with a chemical characterization of macronutrients, minerals by inductively coupled plasma optical emission spectrometry (ICP-OES), phenolic acids and lipid components by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), HPLC-diode-array detector (HPLC-DAD) and gas chromatography-mass spectrometry/flame ionization detector (GC-MS/FID). The study, conducted on different lots of RSM collected over a one-year period from an oil pressing factory serving a biofuel biorefinery, highlighted a constant quality over time of RSM, characterized by high protein (31-34%), fiber (33-40%) and mineral (5.5-6.8%) contents. Polyphenol extracts showed a significant antioxidant activity and a prevalence of sinapic acid, accounting for more than 85% of total phenolic acids (395-437 mg kg-1 RSM). Results highlight the potentialities of RSM for further valorization strategies that may lead to the creation of new cross-sector interconnections and bio-based value chains with improvement of the economics and sustainability of the bioeconomy sectors involved.

Gene editing of three BnITPK genes in tetraploid oilseed rape leads to significant reduction of phytic acid in seeds.

Commercialization of Brassica napus. L (oilseed rape) meal as protein diet is gaining more attention due to its well-balanced amino acid and protein contents. Phytic acid (PA) is a major source of phosphorus in plants but is considered as anti-nutritive for monogastric animals including humans due to its adverse effects on essential mineral absorption. The undigested PA causes eutrophication, which potentially threatens aquatic life. PA accounts to 2-5% in mature seeds of oilseed rape and is synthesized by complex pathways involving multiple enzymes. Breeding polyploids for recessive traits is challenging as gene functions are encoded by several paralogs. Gene redundancy often requires to knock out several gene copies to study their underlying effects. Therefore, we adopted CRISPR-Cas9 mutagenesis to knock out three functional paralogs of BnITPK. We obtained low PA mutants with an increase of free phosphorus in the canola grade spring cultivar Haydn. These mutants could mark an important milestone in rapeseed breeding with an increase in protein value and no adverse effects on oil contents.

Identification of phytic acid mutants in oilseed rape (Brassica napus) by large-scale screening of mutant populations through amplicon sequencing.

Brassica napus (oilseed rape) is an important oil crop in temperate regions, which originated from hybridization of Brassica oleracea and Brassica rapa. Owing to its polyploidy, the functional study of single genes is cumbersome. Phytic acid is considered as an antinutritive compound, and we aimed to knock out the underlying synthesis and transporter genes to identify low phytic acid mutants. We implemented a high-throughput next-generation sequencing screening protocol for an ethylmethane sulfonate population of 7680 plants in six gene families (BnMIPS, BnMIK, Bn2-PGK, BnIPK1, BnIPK2, and BnMRP5) with two paralogues for each gene. A total of 1487 mutations were revealed, and the vast majority (96%) were confirmed by Sanger sequencing. Furthermore, the characterization of double mutants of Bn.2-PGK2 showed a significant reduction of phytic acid contents. We propose to use three-dimensional pooling combined with amplicon stacking and next-generation sequencing to identify mutations in polyploid oilseed rape in a fast and cost-effective manner for complex metabolic pathways. Furthermore, the mutants identified in Bn2-PGK2 might be a very valuable resource for industrial production of oilseed rape protein for human consumption.

Effect of static-state fermentation on volatile composition in rapeseed meal.

BACKGROUND: Fermented rapeseed meal has been used as an alternative protein source for animal feed, but the volatile compounds and how their contents change during fermentation have not been reported. To clarify the effect of static-state fermentation on its aroma, the volatile compounds of rapeseed meal during different stages of fermentation were analyzed using an electronic nose system and headspace solid-phase microextraction-gas chromatography-mass spectrometry. RESULTS: The results suggested that the volatile compounds in the raw rapeseed meal, mostly hydrocarbons and some aldehydes, were lost. The levels of the volatile compounds resulting from microbial metabolism, especially pyrazines, greatly increased during fermentation. Nonanal was the dominant volatile measured in the headspace of raw rapeseed meal. However, the volatile compounds found at high concentrations in rapeseed meal after 5 days of fermentation were tetramethylpyrazine, followed by butanoic acid, benzenepropanenitrile, 2-methylbutanoic acid, trimethylamine, 2,3,5-trimethyl-6-ethylpyrazine, and 2,3,5-trimethylpyrazine. CONCLUSION: The fermentation process could significantly change the composition and content of volatile compounds in rapeseed meal. The results may provide reference data for studies on the choice of fermentation period and formation mechanism of flavor substances in fermented rapeseed meal. © 2020 Society of Chemical Industry.

Effect of supplementation of multiple enzymes to the diets containing variable protein sources on performance and nutrient utilization in commercial broilers.

The effect of supplementation of multiple enzymes at 0, 1X, and 2X concentration to the diet containing variable protein sources (replacement of soybean meal with 10% guar meal (GM), 10% rapeseed meal (RSM), and 10% cottonseed meal (CSM)) was studied in a factorial manner on performance, carcass yield, protein, and energy utilization in commercial broiler males. For this purpose, 600-day-old commercial male broiler chicks (Ross) were randomly divided into 12 treatment groups with 10 replications of 5 birds each and reared in battery brooders up to 42 days of age. Treatment groups are as follows: T1 control and T2 and T3 corn-soya diet with enzymes at 1X and 2X concentrations, respectively. From T4 to T12 treatments, corn-soya meal was replaced by 10% GM, RSM, and CSM without, with 1X and 2X concentration of enzyme supplementation, respectively. The cumulative body weight gain was significantly (P < 0.05) higher in control and birds fed with GM diet in pre-starter and starter as well as during overall period. Supplementation of multiple enzymes at 1X and 2X did not influence body weight, feed intake, and FCR (P > 0.05) during 0-42 days of age. During experimental period, birds fed with the control diet and 10% GM diet showed significantly (P < 0.05) higher cumulative feed intake and better FCR compared to birds fed with 10% RSM diet and 10% cottonseed meal (CSM). Supplementation of multiple enzymes did not influence the mortality and slaughter variables and protein utilization in broilers.

Biotransformation of rapeseed meal leading to production of polymers, biosurfactants, and fodder.

After extracting the oil from rapeseed, the remaining meal byproduct is used in animal feed, particularly for cattle, and represents an effective, high-protein substitute for soybean meal. The biotransformation of rapeseed meal using Generally Recognized as Safe (GRAS) bacteria increases its nutritional value and enriches it with a variety of additives including polymers, biosurfactants, and enzymes. Polymers produced in SSF process with rapeseed meal (e.g., levan) have probiotic prosperities and can even serve as alternatives to antibiotics, which are banned from animal feed by law. Due to their moisturizing properties, these polymers are also incorporated into cosmetics. The biosurfactants produced by bacteria and yeast confer their strong antimicrobial effects to preserve the feed. In turn, the many enzymes produced during the biotransformation of rapeseed meal increase its nutritional value by reducing fibers, detrimental substances (e.g., tannins, erucic acid, phytic acid), and mycotoxins. Taken together, rapeseed meal biotransformation results in numerous benefits, for the animal and industry alike.

The effect of partial substitution of rapeseed meal and faba beans by Spirulina platensis microalgae on milk production, nitrogen utilization, and amino acid metabolism of lactating dairy cows.

Alternative protein sources such as microalgae and faba beans may have environmental benefits over rapeseed. We studied the effects of rapeseed meal (RSM) or faba beans (FB) as a sole protein feed or as protein feeds partially substituted with Spirulina platensis (spirulina) microalgae on milk production, N utilization, and AA metabolism of dairy cows. Eight multiparous Finnish Ayrshire cows (113 ± 36.3 d in milk; mean ± SD) were used in a balanced, replicated 4 × 4 Latin square with 2 × 2 factorial arrangement of treatments and 21-d periods. Four cows in one Latin square were rumen cannulated. Treatments were 2 isonitrogenously fed protein sources, RSM or rolled FB, or one of these sources with half of its crude protein substituted by spirulina (RSM-SPI and FB-SPI). Cows had ad libitum access to total mixed rations consisting of grass silage, barley, sugar beet pulp, minerals, and experimental protein feed. The substitution of RSM with FB did not affect dry matter intake (DMI) but decreased neutral detergent fiber intake and increased the digestibility of other nutrients. Spirulina in the diet decreased DMI and His intake. Spirulina had no effect on Met intake in cows on RSM diets but increased it in those on FB diets. Energy-corrected milk (ECM) and protein yields were decreased when RSM was substituted by FB. Milk and lactose yields were decreased in cows on the RSM-SPI diet compared with the RSM diet but increased in those on FB-SPI compared with FB. The opposite was true for milk fat and protein concentrations; thus, spirulina in the diet did not affect ECM. Feed conversion efficiency (ECM:DMI) increased in cows on FB diets with spirulina, whereas little effect was observed for those on RSM diets. The substitution of RSM by FB decreased arterial concentration of Met and essential AA. Spirulina in the diet increased milk urea N and ruminal NH4-N and decreased the efficiency of N utilization in cows on RSM diets, whereas those on FB diets showed opposite results. Met likely limited milk production in cows on the FB diet as evidenced by the decrease in arterial Met concentration and milk protein yield when RSM was substituted by FB. The results suggest the potential to improve milk production response to faba beans with supplementation of Met-rich feeds such as spirulina. This study also confirmed spirulina had poorer palatability than RSM and FB despite total mixed ration feeding and lower milk production when spirulina partially replaced RSM.