Total Volatile Fatty Acid Concentration Research Articles

63 Impacts of protein supplementation frequency on animal performance, rumen fermentation and methane emission from growing beef cattle

Abstract Nutrient supplementation is one of largest expenditure in forage-based beef production systems, especially when forage quality is poor or limited. Decreasing supplementation frequency may have a role in improving sustainability by increasing animal performance and minimizing cost of labor and fuel. This study evaluated the impacts of protein supplementation frequency (canola meal pellet; CMP) on animal performance, rumen fermentation, and enteric methane production from beef cattle fed forage-based diet. A total of 60 crossbred backgrounding steers [body weight (BW) = 278 ± 10 kg) was randomly assigned to one of the three treatments (n = 20/treatment): i) control (no CMP supplementation), ii) control + everyday CMP supplementation (0.83 kg/d per steer), and iii) control + every other day supplementation of CMP. The study was comprised of 21 d of adaptation followed by three 21-d periods, with enteric methane measured for 7 d per period. Animals were fed grass hay-based diets once a day at 0800 h and remained on their respective diets throughout the 63-d study. Diets were formulated considering animal BW, feed nutrient density and environmental condition in accordance with the National Academies of Sciences, Engineering and Medicine. Each treatment group was assigned to a pen, with the cattle and diets rotated among pens weekly to allow the animals to access the GreenFeed system stationed in one of the pens for enteric methane measurement. For measurement of rumen fermentation products, three ruminally canulated steers (BW = 350 ± 20 kg) were allocated to the three treatments using a 3 x 3 Latin Square Design with 21-d periods so that one animal received all the treatments over the three experimental periods. Animals were adapted to the treatments for 17 d and rumen samples were collected between d 19 and 21 at 0700 h, 1000h, 1300 h, 1600 h and 1900 h. Data were analyzed using the Mixed procedure of SAS. Relative to the control treatment (9.9 kg/d), DM intake was 4 to 5% greater (P < 0.001) following CMP supplementation regardless of supplementation frequency. Similarly, average daily gain was greater (7 to 11%, P = 0.01) for CPM supplemented treatments as compared with control (1.04 kg/d) irrespective of supplantation frequency. Rumen fermentation was affected by treatments. Total VFA concentration was greater (P = 0.01) for the every other day supplementation of CMP as compared with control and daily supplementation treatments. Conversely, rumen ammonia concentration was not affected by treatments. The control group had greater acetate and reduced propionate molar proportions that resulted in a 5 to 6% greater acetate:propionate ratio (P < 0.01) as compared with the CMP supplemented groups. Enteric methane production (g/d) was greater for the daily CMP supplementation (P = 0.02) while yield (g/kg daily weight gain) was not affected by treatment. Our results indicated that reduction of protein supplementation frequency improved animal performance without negatively impacting rumen fermentation.

191 Awardee Talk: Characterizing biological processes influencing the efficiency of nutrient utilization in growing cattle

Abstract 2Abstract: Residual feed intake (RFI), a metric of feed efficiency, is moderately heritable and minimally associated with body size and productivity, making it an ideal trait for investigation as a selection criterion to improve feed efficiency of growing cattle. The objective of this research was to characterize the inter-animal variation in biological processes of growing cattle selected to be divergent in RFI. Holstein heifers [n = 55; body weight (BW) = 351 ± 64 kg) with low- (n = 29) or high- (n = 26) genomically enhanced RFI (RFIg) were selected from a contemporary group of 453 heifers. Heifers were assigned to 1 of 2 pens, equipped with electronic feed bunks and a GreenFeed gaseous-exchange measurement system. Individual dry matter intake (DMI) and feeding behavior data were collected for 84-d. Body weight was measured weekly, and spot fecal samples collected at weighing. Phenotypic RFI (RFIp) was calculated as the residual from regression of DMI on ADG and mid-test BW0.75. On d 70 of the study a rumen fluid sample was collected via esophageal tubing. Blood samples were collected and analyzed for complete blood count (CBC). A mixed model including the fixed effect of RFIg classification and random effect of pen was used to evaluate the effect of RFIg classification on response variables. There were no differences (P > 0.05) in BW and ADG for heifers with divergent RFIg; however, low RFIg heifers consumed 7.5% less (P < 0.05) feed per day. Consequently, low RFIg heifers exhibited a more favorable (P < 0.05) RFIp (-0.188 vs 0.211 kg/d, respectively). Low RFIg heifers exhibited more favorable feeding behavior, with 8.7% fewer (P < 0.05) bunk visit events per day and an 11.2% slower (P ≤ 0.05) eating rate. Low RFIg heifers had 12.5% greater (P < 0.05) total rumen volatile fatty acid concentrations than high RFIg heifers, indicating potentially enhanced microbial fermentation and improved efficiency of feed utilization. Low RFIg heifers had 8.2% less (P < 0.05) methane emissions (g/d), 6.3% less (P < 0.05) carbon dioxide production (g/d), and 6.1% less (P < 0.05) heat production (Mcal/d) calculated according to Brouwer (1965) than high RFIg heifers. Dry matter digestibility did not differ (P > 0.05) between heifers with divergent RFIg. Overall, heifers selected to be more feed efficient exhibited more favorable energy efficiencies and feed efficiency phenotypes. To quantify the variation in RFIg explained by feeding behavior, gas flux, digestibility and rumen fermentation, and CBC partial least squares regression models were developed. Feeding behavior accounted for the largest proportion of variation in RFIg (28.8%). Upon sequential addition of gas flux, digestibility and rumen fermentation, and CBC to feeding behavior, 56.3% of the variation in RFIg was accounted for. Identifying the magnitude at which metabolic processes influence inter-animal variation in efficiency of nutrient utilization provides opportunities to discover valuable candidate biomarkers for selection of more feed efficient cattle.

Effects of Partial Replacement of Corn Grains With Mango Seed Kernel on Nutrient Digestibility, Blood Metabolites and Rumen Parameters of Growing Calves.

Fruit by-products are a sustainable and cost-effective alternative to traditional grain feed for livestock. One of the most important by-products is mango seed kernel (MSK), which can be used as a valuable energy source for feeding growing calves in the tropical and subtropical productive regions of the world. This study investigated the effect of replacing corn grains (CGs) with MSK on intake, nutrient digestibility, blood metabolites and rumen parameters. Eighteen growing male Friesian calves (200 ± 6.55 kg of body weight and 9-11 months old) were randomly assigned to three treatment groups. Three concentrate feed mixtures were formulated such that MSK replaced CG at 0%, 25% and 50%. Dry matter intake was unaffected, whereas dry matter, organic matter and fibre digestibility increased linearly (p < 0.05) with increased inclusion of MSK. Ruminal pH (p = 0.053) and total volatile fatty acid (VFA) concentration (p = 0.041) increased linearly. There was a linear decrease in cholesterol (p = 0.029) and AST (p = 0.028) levels in the blood of calves. In conclusion, this study demonstrated the positive effect of replacing 50% of CG with MSK on rumen parameters, including a higher ruminal pH and total VFA concentration, alongside higher nutrient digestibility. The dietary inclusion of mango seed can serve as a valuable and sustainable dietary component for growing calves.

Metagenomic insights into the mechanistic differences of plant polyphenols and nitrocompounds in reducing methane emissions using the rumen simulation technique

Methane (CH4) emissions from ruminants contribute significantly to greenhouse gas levels and also result in considerable feed energy losses. Plant polyphenols and nitrocompounds are two typical types of methane inhibitors. The study investigates the mechanistic differences between 2-nitroethanol (NE) and proanthocyanidins (PAC) in reducing methane emissions from ruminant livestock using the rumen simulation technique (RUSITEC) combined with metagenomic analyses.The experiment was performed as a complete randomized block design with 3 runs. Run was used as a blocking factor. The treatments included a control (CON) with no additive, NE at 0.5 g/kg dry matter (DM), and PAC at 20 g/kg DM, all incubated in vitro for 24 h (h) with eight replicates per treatment. The results showed that NE significantly reduced CH4 production by 94.9 % (P < 0.01) and total volatile fatty acid (TVFA) concentration by 11.1 % (P < 0.05) compared to the control. NE also decreased the acetate-to-propionate ratio (A/P) from 1.93 to 1.60 (P < 0.01), indicating a shift towards more efficient fermentation. In contrast, PAC reduced methane production by 11.7 % (P < 0.05) and decreased the A/P (P < 0.05) while maintaining microbial diversity and fermentation stability, with no significant impact on TVFA concentration (P > 0.05). Metagenomic analysis revealed that NE markedly suppressed the abundance of key genera involved in carbohydrate metabolism, including Prevotella and Bacteroides, leading to reduced acetate and butyrate pathways. NE also selectively inhibited methanogenic archaea, particularly Methanobrevibacter spp., which are integral to the hydrogenotrophic pathway (P < 0.01). On the other hand, PAC showed selective inhibition of Methanosphaera spp., targeting the methylotrophic pathway (P < 0.01). These findings provide valuable insights into the distinct microbial and metabolic pathways modulated by NE and PAC, offering potential strategies for developing effective dietary interventions to mitigate methane emissions in ruminant livestock.

Effects of Storage Time and Temperature on the Fermentation Characteristics of Rumen Fluid from a High-Forage Diet

The objective of this experiment was to investigate the effects of storage temperature and preservation time on the fermentation characteristics of rumen fluid collected from six Hu sheep fed a high-forage diet. The storage temperatures were set at −80 °C and −20 °C, and the preservation times were labelled as follows: 0 d (fresh rumen fluid, D0), 7 d (D7), 14 d (D14), 30 d (D30), 60 d (D60), 120 d (D120), and 240 d (D240). A repeated-measures design was applied to analyze the fermentation characteristics of rumen fluid across each preservation time point and storage temperature. The results showed that storage temperature had no significant effects on pH value, ammonia nitrogen (NH3-N), microbial protein (MCP), and volatile fatty acid (VFA) concentration and proportion (p &gt; 0.05). Specifically, the pH value increased on D7, D14, and D60 (p &lt; 0.05), while the MCP concentration decreased on D7, D14, D30, D120, and D240 but increased on D60 (p &lt; 0.05), and the concentration of NH3-N decreased on D14 (p &lt; 0.05). The acetate concentration increased on D30, while the concentrations of propionate, butyrate, valerate, and total VFA increased on D60 (p &lt; 0.05). The concentration of isovalerate decreased on D60, and isobutyrate and branched-chain VFA concentrations decreased on D120 (p &lt; 0.05). The proportion of acetate increased on D30, while the butyrate and valerate proportions increased on D60, and the proportions of isovalerate, isobutyrate, and branched-chain VFA decreased on D60 (p &lt; 0.05). For rumen fluid from a high-forage diet, the storage temperature (−80 °C and −20 °C) did not affect its fermentation characteristics, suggesting that rumen fluid could be preserved at −80 °C or −20 °C without altering its fermentation characteristics. The fermentation parameters of rumen fluid changed significantly after 7 days of preservation; hence, it is recommended to finish determining the fermentation parameters of rumen fluid within 7 days of collection. The concentrations and proportions of most VFA changed after 30 days of preservation; therefore, it is recommended that VFA determination should be completed within 30 days of rumen fluid collection.

Effects of Dietary Energy Levels on Growth Performance, Nutrient Digestibility, Rumen Barrier and Microflora in Sheep.

Dietary energy is crucial for ruminants' performance and health. To determine optimal dietary energy levels for growing sheep, we evaluated their growth performance, nutrient digestibility, rumen fermentation, barrier function, and microbiota under varying metabolic energy (ME) diets. Forty-five growing Yunnan semi-fine wool sheep, aged 10 months and weighing 30.8 ± 1.9 kg, were randomly allocated to five treatments, each receiving diets with ME levels of 8.0, 8.6, 9.2, 9.8 or 10.4 MJ/kg. The results showed that with increasing dietary energy, the average daily gain (ADG) as well as the digestibility of dry matter (DM) and organic matter (OM) increased (p < 0.05), while the feed conversion ratio (FCR) decreased linearly (p = 0.01). The concentration of total VFA (p = 0.03) and propionate (p = 0.01) in the rumen increased linearly, while rumen pH (p < 0.01) and the acetate-propionate ratio (p = 0.01) decreased linearly. Meanwhile, the protein contents of Claudin-4, Claudin-7, Occludin and ZO-1 as well as the relative mRNA expression of Claudin-4 and Occludin also increased (p < 0.05). In addition, rumen bacterial diversity decreased with the increase of dietary energy, and the relative abundance of some bacteria (like Saccharofermentans, Prevotella and Succiniclasticum) changed. In conclusion, increasing dietary energy levels enhanced growth performance, nutrient digestibility, rumen fermentation, and barrier function, and altered the rumen bacterial community distribution. The optimal dietary ME for these parameters in sheep at this growth stage was between 9.8 and 10.4 MJ/kg.

Assessing the effects of high-carvacrol oregano oil on rumen microbial fermentation, gas production, and methane production in vitro

The objective of this in vitro study was to assess the effects of high-carvacrol oregano oil (ORE; 50 g/kg oregano oil, 800-802 g/kg carvacrol), on microbial fermentation and CH4 production. In the experiment (a complete randomized block design), treatments included a negative control (CTL, no additive), a positive control (monensin, MON, 10 mg/L), and ORE (20, 40, 80, 120, 200, 160, and 1000 mg/L). Compared with CTL, MON shifted (p &lt; 0.05) volatile fatty acid (VFA) profile from acetate and butyrate to propionate production, thereby reducing (p &lt; 0.05) CH4 production (-26%). Monensin decreased (p &lt; 0.05) NH3 and branched-chain VFA concentrations. Of the doses evaluated, only the highest dose (1000 mg/L) affected ruminal microbial fermentation and CH4 production. At this dose, ORE reduced (p &lt; 0.05) gas production, total VFA, acetate, propionate and NH3 concentrations, and CH4 production (-22%). The reduced gas production and total VFA is an indication of feed digestion inhibition. These results suggest that ORE may decrease CH4 production and improve ruminal N utilization. However, these findings need to be validated in vivo to determine the optimal dose to benefit from the positive effects while avoiding the negative impact of ORE on feed digestion.

Effect of exogenous cellulase pretreatment of straw-type fermented total mixed ration on digestibility and ruminal fermentation in growing beef cattle.

This study aimed to assess the impact of corn straw-based unfermented and fermented total mixed rations (TMR) supplemented with exogenous cellulase on the in vitro fermentation characteristics, growth performance, feeding behavior, apparent digestibility, rumen fermentation and digestive enzyme activities of Chinese Simmental bulls. Unfermented (direct spraying of exogenous cellulase onto TMR, TMR) and fermented (exogenous cellulase fermentation for more than 7 d, fermented total mixed rations [FTMR]) TMR were collected, dried, powdered and used as fermentation substrates. The fermentation liquid was ruminal fluid collected from Chinese Simmental bulls. The artificial rumen culture fluid were continuously cultured in vitro for 48 h. Based on the diets they were fed, 24 healthy Chinese Simmental bulls (average weight of 495.93 ±10.89 kg) were randomly divided into two groups, with 12 bulls in each group, which were fed TMR or FTMR. The study lasted 56 d. In in vitro experiments, the neutral detergent fiber degradability and total volatile fatty acid, propionate, iso-butyrate, iso-valerate and valerate concentrations were greater in the FTMR group (p<0.05) than in the TMR group. However, the methane production, pH and A/P of the FTMR group tended to be lower (p<0.05) than those of the TMR group. In the in vivo experiments, the average daily gain, eating rate, and feed efficiency of the FTMR groups were greater (p<0.05) than those of the TMR group. Similarly, the NDF degradability of the FTMR group was greater (p<0.05) than that of the TMR group. Compared to those in the TMR group, the concentrations of total volatile fatty acids, iso-butyrate, propionate and butyrate were greater in the FTMR group (p<0.05), and the A/P ratio was lower (p<0.05). Similarly, cellulase, xylanase, and β-glucosidase activities were greater (p<0.05) in the FTMR group than in the TMR group. Corn straw-based fermented total mixed rations supplemented with exogenous cellulase play a vital role in decreasing the structural carbohydrate content of TMR and ruminal methane production in vitro, improving nutrient digestion and absorption, optimizing rumen fermentation, and improving the growth performance of beef cattle.

Volatile Fatty Acids from Household Food Waste: Production and Kinetics

Household food waste (HFW), which is rich in organic matter, is a good candidate for producing added-value bio-based chemicals, such as volatile fatty acids (VFAs), by acidogenic fermentation processes. However, the lack of design tools, such as appropriate kinetic models, hinders the implementation of this technology because the results of these processes are affected by operational factors. In this work, VFA production by the acidogenic fermentation of HFW under uncontrolled pH levels (4–5) was studied at thermophilic (55 °C) and mesophilic (35 °C) temperature conditions. Batch reactors were used to digest HFW, and VFA production and the individual acid distributions were measured at different fermentation times from 0 to 624 h. The results showed higher individual and total VFA production at 35 °C and 120 h of fermentation time as a consequence of the competition between the VFA production and decomposition reactions. Acetic and valeric acids were VFAs mainly produced as a result of a high content of proteins in the initial substrate, and a small amount of propionic and butyric acids were present. A simplified kinetic model was successfully developed to represent the complex process of VFA formation from the acidogenic fermentation of HFW. A simple mechanism for the production–decomposition of VFAs, corresponding to a zero-order reaction for the first 48 h and a single consecutive reaction from that time on, was proposed. For both mesophilic and thermophilic conditions, the suggested kinetic model was able to predict the individual and total concentrations of VFAs along the fermentation time.

Feeding almond hulls to light lambs: Effects on growth performance, digestive utilization and blood metabolites

The effects of including almond hulls (AH) in the concentrate fed to light lambs on animal performance, diet utilization, and blood metabolites. Three isonitrogenous and isoenergetic concentrates were formulated with either 0 (control), 6 (AH6) or 12 (AH12) g of AH per 100 g (fresh matter basis). Thirty Manchega breed lambs (15 males and 15 females; 12.3 ± 0.18 kg body weight (BW)) were distributed into six homogeneous groups according to BW (3 groups of each sex), which were randomly assigned within sex to each experimental concentrate. Lambs were fed concentrate and barley straw ad libitum until reaching about 23.0 kg BW. There were no differences (P ≥ 0.182) between groups in final BW, intake of concentrate and straw, and feed conversion rate. Average daily gain and digestibility of dry matter and crude protein tended (P < 0.10) to decrease by the AH inclusion in the concentrate. Lambs fed AH12 showed greater (P < 0.05) urinary losses than those fed the control concentrate (15.4 and 11.5 g per 100 g of N intake, respectively), despite the similar N intake in all groups. Feeding AH did not affect (P ≥ 0.213) blood concentrations of albumin, globulins, urea, creatinine, glucose, total bilirubin and cholesterol or blood activities of hepatic enzymes. The post-mortem analysis revealed no differences among groups in pH and total volatile fatty acids (VFA) concentrations in the rumen of lambs, but NH3-N concentrations were lower (P < 0.05) in the lambs fed the concentrates with AH. In addition, lambs fed AH12 diet had lower (P < 0.05) molar proportions of propionate and greater (P < 0.05) proportions of butyrate compared with lambs fed control and AH6 concentrate. In general, no concentrate x sex interactions were observed, and sex had no effect on the measured parameters except for some differences in ruminal VFA profile. In conclusion, AH can be included at in light lambs’ concentrate up to 12 g per 100 g of concentrate without impairing animal growth.

Effects of an essential oil blend rich in cinnamaldehyde and carvacrol on rumen biohydrogenation and fatty acid profile in the longissimus lumborum of growing lambs.

Essential oils extracted from cinnamon bark and oregano are rich in cinnamaldehyde and carvacrol and show potential for promoting animal performance. However, their impact on rumen biohydrogenation and the fatty acid composition of meat has not been reported. The hypothesis of this study was that a blend of essential oils rich in cinnamaldehyde and carvacrol would inhibit rumen biohydrogenation and promote the accumulation of polyunsaturated fatty acids (PUFAs) in lamb meat. The present study evaluated the effect of a blend essential oil (EO) rich in cinnamaldehyde and carvacrol on the nutrient digestibility, rumen biohydrogenation, growth performance, and fatty acid profile of the longissimus lumborum of lambs. Sixty male lambs with an average age of 84 ± 0.98 days and initial body mass of 25.4 ± 0.29 kg (mean ± standard deviation) were assigned randomly to four diets, and supplemented with 0 (EO0), 30 (EO30), 60 (EO60), and 120 (EO120) mg kg-1 dry matter of EO for 60 days. Although dry matter and neutral detergent fiber digestibility all showed a linear decrease (P ≤ 0.02) with increasing quantities of EO, final body mass and average daily gain increased linearly (P = 0.04), and average daily weight gain (ADG)/dry matter intake (DMI) tended to increase linearly (P = 0.07). Increasing EO supplementation resulted in a linear decrease in total volatile fatty acid concentration, acetate molar percentage, and acetate-to-propionate ratio (P ≤ 0.03), with the EO120 treatment being lower than the other EO treatments (P ≤ 0.05). Seven lambs from the EO120 treatment and seven lambs from the EO0 treatment were randomly slaughtered. It was observed that the proportions of C18:2n6c and PUFA in longissimus lumborum were higher in the EO120 treatment than the EO0 treatment (P ≤ 0.05). The relative abundance of Firmicutes in the rumen was decreased by the EO120 treatment in comparison with the EO0 treatment (P ≤ 0.05). Furthermore, the predicted relative abundances of genes encoding for conjugated linoleic acid reductase tended to decrease with the EO120 treatment (P = 0.06). We demonstrated that supplementation of the EO blend rich in cinnamaldehyde and carvacrol can enhance lamb growth performance and promote the deposition of desirable PUFAs in meat. © 2024 Society of Chemical Industry.

Effects of yeast culture supplementation on milk yield, rumen fermentation, metabolism, and bacterial composition in dairy goats.

The effects of yeast culture (YC) on dairy goat milk yield and potential effects of rumen microbial population changes on rumen fermentation are poorly understood. This study aimed to evaluate the effects of YC on milk yield and rumen fermentation in dairy goats and explore the potential microbial mechanisms. Forty Laoshan dairy goats with a weight of 51.23 ± 2.23 kg and daily milk yield of 1.41 ± 0.26 kg were randomly divided into 4 groups: control (no YC), YC1 (10 g/day per goat), YC2 (25 g/day per goat), and YC3 (40 g/day per goat). The pre-feeding period was 15 days, and the official period was 60 days. Laoshan dairy goats were milked twice daily, and the individual milk yield was recorded. On the last day of the official period, rumen fluid was collected to measure rumen fermentation, perform quantitative polymerase chain reaction (PCR), and detect metabolites. Compared to the control group, the YC group had greater milk yield; higher acetic acid, butyric acid, and total volatile fatty acid contents; and lower ammonia-N (NH3-N) content in the rumen (p < 0.05). YC increased the abundance of Clostridia_UCG-014 and Paraprevotella (p < 0.05). Differential metabolites L-leucine and aspartic acid were screened. This study revealed the microbial mechanisms linking the relative abundance of Paraprevotella and Clostridia_UCG-014 to L-leucine and aspartic acid utilization. These results describe the potential benefits of supplementing 10 g/day per goat YC in the diets of Laoshan dairy goats for improving the rumen environment and milk yield.

Effects of fermented jujube powder on growth performance, rumen fermentation, and antioxidant properties of simmental bulls.

Fermented jujube powder (FJP) promotes a balance between the intestinal microflora and immune factors in animals. In this study, we aimed to investigate the effects of FJP on the production performance, nutrient digestion, rumen fermentation, and antioxidant properties of bulls. Forty Simmental bulls were randomly divided into four groups based on body weight and fed a basal diet with [5, 7.5, or 10% dry matter (DM)] or without FJP. The experimental period was 20 d for adaptation and 60 d for the feeding trial. Dietary FJP supplementation did not affect DM intake (P > 0.05) but increased the average daily gain quadratically (P = 0.049) and decreased the feed conversion ratio linearly (P = 0.042). FJP quadratically enhanced DM and crude protein digestibility (P = 0.026 and P = 0.041, respectively) and linearly enhanced acid detergent fiber digestibility (P = 0.048). It also increased the total volatile fatty acid concentration quadratically (P = 0.037), acetate molar percentage, and acetate-to-propionate ratio linearly (P = 0.002 and 0.001), and reduced the ammonia nitrogen concentration linearly (P = 0.003). Additionally, xylanase and protease activities and Ruminococcus flavefaciens abundance increased linearly (P = 0.006, 0.018, and 0.009, respectively), and total bacteria, Ruminococcus albus, and Ruminobacter amylophilus abundance increased quadratically (P = 0.047, 0.011, and 0.021, respectively). FJP linearly increased serum total protein concentration and antioxidant capacity (P = 0.003 and 0.018, respectively) and decreased malonaldehyde content (P = 0.006). FJP supplementation (7.5%) enhanced production performance, nutrient digestion, rumen fermentation, and serum antioxidant capacity in bulls. The improved nutrient digestion may be due to an increase in ruminal microorganisms and total volatile fatty acids from the FJP. High blood antioxidant levels indicate that FJP may preserve proteins, thereby boosting the production performance of bulls.

Digestion, nitrogen excretion and rumen microbial communities in beef cattle, and in vitro methane output, of fresh temperate grass with or without citrus pulp supplementation

The effects of supplementing grass with a high-energy, low-protein concentrate on intake, rumen fermentation, rumen microbial communities, apparent total tract digestibility and N-balance in beef cattle, and rumen fermentation and methane output in vitro, were studied. Sixteen Charolais steers (mean live weight (s.d.) 564 (24.3) kg)), were used in a completely randomised block design experiment, and offered either zero-grazed perennial ryegrass (Lolium perenne L.) ad libitum (GO), or the same grass with 2.25 kg citrus pulp dry matter (DM) daily (GCP). During the measurement phase, DM intake was restricted at 0.85 of their pre-determined intake. The same diets were incubated in an eight-vessel RUSITEC system. Supplementation decreased grass DM intake (6.59 vs. 7.54 kg/d) but increased total DM intake by 0.95 kg/d (P < 0.01). Rumen pH (P = 0.05) and NH3 concentration (P < 0.05) decreased, and total volatile fatty acid (VFA) concentrations (P < 0.01), molar proportions of propionate (P = 0.08) and butyrate (P < 0.05) increased for GCP compared to GO. Supplementation significantly altered the overall rumen microbial community structure compared to GO, driven principally by changes in relative abundance of Prevotella, Methoanobrevibacter and Lachnospiraceae genera. Plasma urea concentration was lower (P < 0.05) for GCP than GO. There was no effect (P > 0.10) of supplementation on apparent total tract digestibility of DM, OM and ADFom, but aNDFom digestibility tended to be lower (P = 0.07) and crude protein (N × 6.25) digestibility was lower (P < 0.01) for GCP compared to GO. Nitrogen intake (200 g/d) did not differ (P > 0.05) between treatments. Faeces N loss was 9 g/d greater (P < 0.01), and urine (P < 0.01) and total (P < 0.05) N losses were 22 and 13 g/d lower, respectively, for GCP compared to GO. In terms of N-balance (g/kg N intake), faecal N excretion was greater and urinary N excretion was lower (P < 0.01) resulting in a greater N use efficiency (P = 0.06) for GCP than GO. In vitro rumen NH3 concentrations were lower (P < 0.01) for GCP compared to GO, whereas in vitro rumen pH, lactic acid, VFA concentrations and molar proportions, and methane and total gas output did not differ (P > 0.05) between treatments. Supplementing grass with a low-protein, high-energy feed may be a strategy to reduce urinary and total N excretion in beef cattle grazing temperate grass pasture.

Pilot-scale evaluation of cascade anaerobic digestion of mixed municipal wastewater treatment sludges.

This work assessed the performance of a pilot-scale cascade anaerobic digestion (AD) system when treating mixed municipal wastewater treatment sludges. The cascade system was compared with a conventional continuous stirred tank reactor (CSTR) digester (control) in terms of process performance, stability, and digestate quality. The results showed that the cascade system achieved higher volatile solids removal (VSR) efficiencies (28-48%) than that of the reference (25-41%) when operated at the same solids residence time (SRT) in the range of 11-15 days. When the SRT of the cascade system was reduced to 8days the VSR (32-36%) was only slightly less than that of the reference digester that was operated at a 15-day SRT (39-43%). Specific hydrolysis rates in the first stage of the cascade system were 66-152% higher than those of the reference. Additionally, the cascade system exhibited relatively stable effluent concentrations of volatile fatty acids (VFAs: 100-120 mg/l), while the corresponding concentrations in the control effluent demonstrated greater fluctuations (100-160 mg/l). The cascade system's effluent pH and VFA/alkalinity ratios were consistently maintained within the optimal range. During a dynamic test when the feed total solids concentration was doubled, total VFA concentrations (85-120 mg/l) in the cascade system were noticeably less than those (100-170 mg/l) of the control, while the pH and VFA/alkalinity levels remained in a stable range. The cascade system achieved higher total solids (TS) content in the dewatered digestate (19.4-26.8%) than the control (17.4-22.1%), and E. coli log reductions (2.0-4.1 log MPN/g TS) were considerably higher (p < 0.05) than those in the control (1.3-2.9 log MPN/g TS). Overall, operating multiple CSTRs in cascade mode at typical SRTs and mixed sludge ratios enhanced the performance, stability digesters, and digestate quality of AD. PRACTITIONER POINTS: Enhanced digestion of mixed sludge digestion with cascade system. Increased hydrolysis rates in the cascade system compared to a reference CSTR. More stable conditions for methanogen growth at both steady and dynamic states. Improved dewaterability and E. coli reduction of digestate from the cascade system.

Impact of aging of primary and secondary polystyrene nanoplastics on the transmission of antibiotic resistance genes in anaerobic digestion

Sewage sludge is a significant reservoir of nano/microplastics (NPs/MPs) and antibiotic resistance genes (ARGs). Research has revealed that NPs/MPs may exert an inhibitory effect on anaerobic digestion (AD) of sludge. Moreover, NPs/MPs can influence microbial community diversity and composition, potentially increasing ARGs dissemination. The morphological changes to NPs/MPs surface due to aging contribute to modifying hydrophobic properties. To date, there is limited comprehension regarding how various surface properties of NPs influence ARGs dissemination during AD. This study investigated the impact of primary aged/non-aged and secondary aged/non-aged polystyrene nanoplastics (PSNPs) on ARGs and mobile genetic elements (MGEs) propagation during AD. The findings indicated that the UV-aging process resulted in surface oxidation and distinct morphological characteristics in both primary and secondary PSNPs, while the surface oxidation effect was more pronounced in the secondary aged PSNPs. High concentrations (150 μg/L) of primary and secondary PSNPs inhibited methane production, with secondary PSNPs causing greater inhibition by 16 to 20 % compared to control. In contrast, low concentration (25 μg/L) had negligible or slightly positive effects on methane production. PSNPs at 150 μg/L reduced total VFA concentration, indicating an inhibitory effect on the fermentation step in the AD process. Primary and secondary PSNPs exhibited changes in EPS characteristics. ARGs abundance was enriched in reactors amended with PSNPs, with the highest abundance of 8.54 × 105 copies/g sludge observed in the secondary aged PSNPs (150 μg/L) reactor. Reactors exposed to aged PSNPs exhibited a relatively higher abundance of ARGs compared to reactors exposed to non-aged PSNPs. Exposure to PSNPs increased the microbial community diversity within the digesters and triggered the enrichment of Comamonadaceae and Syntrophaceae, belonging to Proteobacteria phylum. On the other hand, archaeal communities tended to shift towards hydrogenotrophic methanogens in PSNPs reactors. The correlation analysis showed that Comamonadaceae were positively correlated with the majority of ARGs and intl1. A positive correlation was observed between MGEs and most ARGs, suggesting that the increased proliferation of ARGs under PSNPs exposure may be linked to the abundance of MGEs, which in turn promotes the growth of hosts carrying ARGs. These findings suggest that aged and non-aged NPs could substantially impact the spread of ARGs and MGEs, which also led to notable alterations in the composition of the microbial community. Overall, this study provides valuable insights into the multifaceted impacts of PSNPs with various characteristics on AD processes, microbial communities, and ARGs proliferation, highlighting the urgent need for comprehensive assessments of NPs pollutants in the environment.

Effects of sodium chloride intake on urea-N recycling and renal urea-N kinetics in lactating Holstein cows

The effects of high (2.5% of DM) versus normal dietary sodium chloride (NaCl) intake on renal urea-N kinetics and urea-N metabolism were investigated in 9 rumen-cannulated and multi-catheterized lactating dairy cows in a crossover design with 21-d periods. It was hypothesized that urinary urea-N excretion would be greater, and blood urea-N concentration lower in response to greater diuresis induced by high NaCl intake. Also, urea-N transport across ruminal and portal drained viscera (PDV) tissues was hypothesized to be affected by dietary sodium intake. A second experiment was conducted using 8 lactating cows in a crossover design with 14-d periods to test high NaCl (2.5% of DM) versus high KCl (3.2% of DM) intake on milk yield and milk urea-N concentrations. Experiment 1 showed that despite greater diuresis there was no effect of high NaCl intake on urinary urea-N excretion or blood urea-N concentration. The high NaCl intake did not affect rumen ammonia concentrations, total rumen VFA concentrations, ruminal venous - arterial concentration differences for ammonia, or ammonia absorption indicating that high NaCl did not adversely affect ruminal fermentation and microbial protein synthesis. High NaCl intake did not affect the total amount of urea-N transport from blood to gut, but ruminal venous - arterial concentration differences for urea-N were lower with high NaCl and ruminal extraction of arterial urea-N was numerically smaller, indicating that the ruminal epithelial urea-N transport was lower with high NaCl. Energy corrected milk yield was greater with high NaCl (3.2 ± 1.5 kg/d); however, milk urea-N concentrations were not affected by treatment. In experiment 2, ECM was greater with NaCl (1.4 ± 0.31 kg/d) compared with KCl (30.2 and 28.8 ± 0.91 kg ECM / d, respectively). Milk urea-N concentration was lower with KCl, suggesting a urea-N lowering effect in milk not evident with high NaCl intake. In conclusion, the present data show that dietary Na intake of 12-13 g/kg DM was followed by greater diuresis but did not impact urea-N excretion or blood urea-N concentration. High NaCl intake did not affect the total amount of urea-N transfer across PDV tissues. Energy corrected milk yield was greater with high NaCl compared with both control and feeding KCl, however, with KCl milk urea-N decreased.

Performance, interaction, and metabolic pathway of novel dry–wet anaerobic digestion for treating high-solid agricultural waste

Anaerobic digestion (AD) with high-solid content of agricultural waste often leads to the accumulation of volatile fatty acids (VFAs) due to the imbalance of the functions of hydrolytic acidification bacteria and methanogenic archaea. This study developed a novel dry–wet AD of high-solid content was developed to make acidogenic bacteria and methanogenic archaea having higher collaboration ability. The results showed that the dry–wet AD system achieved a dynamic balance and synergistic effect between hydrolytic acidification and the methane production process, which shortened the lag phase to a minimum of 1.21 days. Additionally, the system exhibited a highest volumetric biogas production rate of 1.76 L L−1 d−1 when CM:CS and the initial pH were 6:4, 8.0, respectively. The total VFA concentration reached a maximum of 4.69 g L−1 d−1 and acetic acid (HAc) and butyric acid (HBu) were dominant VFAs (88.75%–93.27%) in the leachate in hydrolytic acidification phase. Metagenomic analysis indicated Pseudomonadales, Clostridiales, Bacteroidales, Spirochaetales, and Methanosaeta were dominant microorganism in hydrolytic acidification phase and methanogenic phase, respectively. The gene abundance of key enzymes in the acetoclastic methanogenic pathway significantly exceeded that of the hydrotrophic methanogenic pathway. This study provides a novel AD technology and insights into the energy conversion of agricultural waste.

Effect of acidosis in the late-finishing phase on rumen fermentation in feedlot steers.

The objective was to determine the effects of induced acidosis in the late-finishing phase on rumen fermentation in feedlot steers. Eleven ruminally cannulated steers (body weight [BW] = 795 kg ± 54) were blocked into two groups based on initial BW. For 195 d prior to the start of the study, cattle were consuming a basal finishing diet (60% dry-rolled corn, 15% modified distillers grains, 15% corn silage, and 10% ground corn-based supplement). Steers were randomly assigned to one of the two treatments: control (CON), or induced acidosis (ACD). Both treatments were fasted for 24h then fed the basal finishing diet. Steers on the ACD treatment received 0.05% of BW of wheat starch via rumen cannula at 0800 and 2000 hours on day 1 and ad libitum refeeding following the fast. On days 1 and 2, CON steers were provided 25% of allotted feed every 6h. Rumen fluid was collected every 4h during the challenge period (hours 0 to 48), and 0, 6, and 12h after feeding during the recovery period (hours 54 to 96). Rumen fluid was analyzed for pH, ammonia, volatile fatty acids (VFA), and lactate. Fecal grab samples were collected every 8h to determine fecal pH. A treatment × day interaction (P = 0.03) was observed for dry matter intake during the challenge period with steers on the ACD treatments consuming more on day 1 than CON steers. Intake was not different on day 2 (P = 0.88). A treatment × hour effect (P < 0.01) was observed for ruminal pH during the challenge period with the ACD steers having a lesser pH than CON from hours 12 to 32. Duration of time below a pH of 5.6 during the challenge period was greater (P < 0.01) for ACD steers than CON. During the challenge period, a treatment × time interaction (P = 0.04) was observed for total VFA concentration with ACD steers having greater total VFA concentration from hours 12 to 36. Acetate to propionate ratio (A:P) was affected by treatment × hour (P = 0.04) with CON steers having greater A:P from hours 28 to 48. Rumen ammonia and lactate concentrations did not differ (P ≥ 0.25) between treatments or the interaction with time. Challenge and recovery period fecal pH were not affected (P ≥ 0.13) by treatment, time, or their interaction. Recovery period ruminal pH was not different (P = 0.99) between treatments. For the recovery period, total VFA and ammonia concentration were not affected by treatment, time, or their interaction (P ≥ 0.07). Ruminal pH and VFA were affected in the initial 48h of induced acidosis in the late-finishing phase.

Supplementing flaxseed meal with sucrose, flaxseed oil, or both: Effects on milk enterolactone, ruminal microbiota profile, production performance, and nutrient utilization in dairy cows

Flaxseed is the richest source of secoisolariciresinol diglucoside, which is converted by ruminal microorganisms primarily to the mammalian lignan enterolactone. Our objective was to investigate the effect of diets containing soybean meal or flaxseed meal (FM) supplemented with sucrose, flaxseed oil, or both, on milk enterolactone concentration yield, diversity, and relative abundance of ruminal bacterial taxa, ruminal fermentation profile, production performance, milk fatty acids (FA) yield, and nutrient utilization in dairy cows. Sixteen Holstein cows (8 multiparous [4 ruminally cannulated] and 8 primiparous) averaging (mean ± SD) 134 ± 54.1 DIM and 679 ± 78.9 kg of BW in the beginning of the study were assigned to treatment sequences in a replicated 4 × 4 Latin square design. Each experimental period lasted 25 d, with 18 d for diet adaptation and 7 d for data and sample collection. Diets were formulated to contain a 60:40 forage:concentrate ratio and included the following (DM basis): (1) 8% soybean meal and 23% ground corn (SBM); (2) 15% FM, 10.7% ground corn, and 5% sucrose (FLX+S); (3) 15% FM, 15.4% ground corn, and 3% flaxseed oil (FLX+O); and (4) 15% FM, 10.2% ground corn, 5% sucrose, and 3% flaxseed oil (FLX+SO). Compared with SBM, the concentration and yield of milk enterolactone increased in cows fed the FM diets but did not differ among FLX+S, FLX+O, and FLX+SO. The relative abundances of the phyla Firmicutes, Verrucomicrobiota, and Actinobacteriota, and those of the bacterial genera Lachnospiraceae NK3A20 group, Eubacterium coprostanoligenes group, Anaeromusa-Anaeroarcus, WCHB1-41, and p-251-o5 decreased, whereas Prevotella and NK4A214 group increased when comparing SBM against at least 1 diet containing FM. Furthermore, the relative abundances of Firmicutes and Actinobacteriota and those of Prevotella, Lachnospiraceae NK3A20 group, Eubacterium coprostanoligenes group, Acetitomaculum, Lachnospiraceae unclassified, NK4A214 group, and Anaeromusa-Anaeroarcus changed (increased or decreased) across the FLX+S, FLX+O, and FLX+SO diets. However, all these changes in the relative abundance of the ruminal bacterial taxa were not conclusively associated with the effect of diets on milk enterolactone. Diets did not affect ruminal pH and concentrations of NH3-N and total VFA. Dry matter intake and yields of milk, milk fat, and milk true protein all decreased in cows fed FLX+O or FLX+SO. Yields of milk total odd-chain FA, branched-chain FA, total <16C FA, and total 16C FA decreased with feeding FLX+O and FLX+SO. The apparent total-tract digestibilities of DM and OM were lowest in the FLX+S and FLX+O diets, with CP and ADF digestibilities lowest in cows receiving FLX+S or FLX+O, respectively. Urinary excretion of total N was lowest with feeding SBM. Contrarily, diets did not affect the urinary excretion of total purine derivatives. In brief, despite the effect of diets on the relative abundance of several ruminal microbiota phyla and genera, we were unable to conclusively associate these changes with increased milk enterolactone in FM-containing diets versus SBM.