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This study aimed to evaluate the sulfur hexafluoride (SF6) tracer technique for estimating methane (CH4) emissions in dual-flow continuous culture systems (DFCCS). In experiment 1 (Exp1), fermenters were filled with water, and known CH4 concentrations (0, 1.35, 2.93, or 4.43 g/d) were injected using permeation tubes with SF6 release rates (3.30 or 9.65 mg/d). Headspace gas was collected using canisters, and the SF6 technique estimated CH4 recovery. Experiment 2 (Exp2) involved a DFCCS fermentation trial with ruminal fluid from three Holstein cows, testing diets with soybean meal or its partial replacement (50%) by Chlorella or Spirulina. Headspace gas was collected at intervals post-feeding. Standard curves for SF6 and CH4 quantification were inadequate for DFCCS samples, with the CH4:SF6 ratio differing from standards, indicating the data needs further SF6 release rate evaluation. In Exp1, a high correlation (r = 0.97) was found between infused and calculated CH4, indicating good repeatability. Low and high SF6 rates performed similarly at low CH4 infusion, but high SF6 overestimated CH4 at high infusion. Exp2 showed CH4 emissions irrespective of SF6 rate and indicated reduced CH4 emissions and increased NDF degradation with algae-containing diets. Further evaluation of the SF6 tracer technique is warranted for DFCCS. [ABSTRACT FROM AUTHOR]

Aflatoxin B1 (AFB1) is a mycotoxin known to impair human and animal health. It is also believed to have a deleterious effect on ruminal nutrient digestibility under in vitro batch culture systems. The objective of this study was to evaluate the effects of increasing the dose of AFB1 on ruminal dry matter and nutrient digestibility, fermentation profile, and N flows using a dual-flow continuous culture system fed a diet formulated for lactating dairy cows. Eight fermenter vessels were used in a replicated 4 × 4 Latin square design with 10 d periods (7 d adaptation and 3 d sample collection). Treatments were randomly applied to fermenters on diet DM basis: (1) 0 μg of AFB1/kg of DM (Control); (2) 50 μg of AFB1/kg of DM (AF50); (3) 100 μg of AFB1/kg of DM (AF100); and (4) 150 μg of AFB1/kg of DM (AF150). Treatments did not affect nutrient digestibility, fermentation, and N flows. Aflatoxin B1 concentration in ruminal fluid increased with dose but decreased to undetectable levels after 4 h post-dosing. In conclusion, adding incremental doses of AFB1 did not affect ruminal fermentation, digestibility of nutrients, and N flows in a dual-flow continuous culture system fed diets formulated for lactating dairy cows. [ABSTRACT FROM AUTHOR]

The utilization of microencapsulated organic acids and pure botanicals (mOAPB) is widely used in the monogastric livestock industry as an alternative to antibiotics; in addition, it can have gut immunomodulatory functions. More recently, an interest in applying those compounds in the ruminant industry has increased; thus, we evaluated the effects of mOAPB on ruminal fermentation kinetics and metabolite production in an in vitro dual-flow continuous-culture system. For this study, two ruminal cannulated lactating dairy Holstein cows were used as ruminal content donors, and the inoculum was incubated in eight fermenters arranged in a 4 × 4 Latin square design. The basal diet was formulated to meet the nutritional requirements of a 680-kg Holstein dairy cow producing 45 kg/d of milk and supplemented with increasing levels of mOAPB (0; 0.12; 0.24; or 0.36% of dry matter [ DM ]), which contained 55.6% hydrogenated and refined palm oil, 25% citric acid, 16.7% sorbic acid, 1.7% thymol, and 1% vanillin. Diet had 16.1 CP, 30.9 neutral detergent fiber (NDF), and 32.0 starch, % of DM basis, and fermenters were fed 106 g/d split into two feedings. After a 7 d adaptation, samples were collected for 3 d in each period. Samples of the ruminal content from the fermenters were collected at 0, 1, 2, 4, 6, and 8 h postmorning feeding for evaluation of the ruminal fermentation kinetics. For the evaluation of the daily production of total metabolites and for the evaluation of nutrient degradability, samples from the effluent containers were collected daily at days 8 to 10. The statistical analysis was conducted using MIXED procedure of SAS and treatment, time, and its interactions were considered as fixed effects and day, Latin square, and fermenter as random effects. To depict the treatment effects, orthogonal contrasts were used (linear and quadratic). The supplementation of mOAPB had no major effects on the ruminal fermentation, metabolite production, and degradability of nutrients. The lack of statistical differences between control and supplemented fermenters indicates effective ruminal protection and minor ruminal effects of the active compounds. This could be attributed to the range of daily variation of pH, which ranged from 5.98 to 6.45. The pH can play a major role in the solubilization of lipid coat. It can be concluded that mOAPB did not affect the ruminal fermentation, metabolite production, and degradability of dietary nutrients using an in vitro rumen simulator. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effects of feeding sugars as a replacement for starch on the ruminal microbiome using a dual-flow continuous culture system. Four periods of 10 days each were conducted with 8 fermenters in a 4 × 4 replicated Latin square design. Treatments included: 1) control with corn—CON, 2) molasses—MOL, 3) untreated condensed whey permeate—CWP, and 4) CWP treated with a caustic agent—TCWP as a partial substitute for corn. Sugars were defined as the water-soluble carbohydrates (WSC) concentration. Diets were formulated by replacing 4% of the diet DM in the form of starch from corn with the sugars in byproducts. Microbial samples for DNA analysis were collected from the solid and liquid effluent containers at 3, 6, and 9 h after feeding. Bacterial community composition was analyzed with sequencing the V4 region of the 16S rRNA gene using Illumina MiSeq platform. Data were analyzed with R 4.1.3 packages vegan, lmer, and ggplot to determine the effects of treatment on the relative abundance of taxa in the solid and liquid fractions, as well as the correlation of Acetate: Propionate ratio and pH to taxa relative abundance. Treatments did not affect alpha or beta diversity. At the phylum level the relative abundance of Proteobacteria was increased in CON compared to sugars in the solid fraction. In the liquid fraction, Firmicutes had greater relative abundance in sugar treatments while Bacteroidota and Spirochaetota were present in lower relative abundance in CWP. For solid and liquid samples, the family Lachnospiraceae had greater relative abundance in sugar treatments compared to CON. The decreased relative abundance of Christensenellaceae and Rikenellaceae paired with the greater relative abundance of Selenomonadaceae in CWP could help explain greater propionate molar proportion and decreased ruminal pH previously observed for this treatment. The genera Olsenella a lactic acid-producing bacterium, had the greatest relative abundance in MOL. Incorporating TCWP or MOL as a partial replacement for starch was more conservative of fibrolytic bacterial taxa compared to CWP. Additionally, TCWP did not increase bacterial taxa associated with synthesis of lactate as compared to MOL. Overall, replacing starch with sugars is mostly conservative of the ruminal microbiome; however, changes observed coincide with differences observed in acetate and propionate proportions and ruminal pH. [ABSTRACT FROM AUTHOR]

Climate change may have an adverse effect on feed production. Hydroponic fodders produced in controlled environments are alternative feeds that may be used for dairy cattle. The objective of this study was to evaluate in-situ dry matter ruminal degradability of hydroponic fodders and corn silage. Two rumen-canulated lactating Holstein cows were used in a complete randomized block design with four treatments and 9 times (0, 2, 4, 8, 12, 24, 48, 72, and 240 h). Treatments were corn silage (CS), hydroponic sprouts of barley, wheat, and triticale. Quadruplicate samples were placed in Dacron bags (10 x 20 cm, 50 μm porosity) and incubated in the rumen. After incubation, samples were washed and dried in an air-forced oven set at 60°C for 48 h and DM recovery was weighted. Ingredients nutrient composition were CS (32.3% NDF, 8.4% CP, and 41.6% starch, 1.3% WSC; % of DM), barley (34.3% NDF, 17.1% CP, and 4.2% starch, 32.9% WSC; % of DM), wheat (23.7% NDF, 23.3% CP, and 8.5% starch, 32.7% WSC; % of DM), triticale (20.0% NDF, 16.2% CP, and 21.7% starch, 33.2% WSC; % of DM). Model included fixed effects of treatment and random effects of cow and replicate(cow). Data were analyzed using PROC MIXED of SAS. Degradation fractions were calculated based on DM disappearance. There were treatment effects (P < 0.01) on fraction A (degradable), fraction B (potentially degradable), fraction C (undegradable), effective ruminal degradability (ERD), and rate of degradation (kd: %/h). Fraction A was greater for triticale (74.1%) compared with wheat (70.2%), CS (63.1%), and barley (55.0%). Fraction B was greater for barley (28.5%) compared with CS (23.2%), wheat (19.1%), and triticale (13.9%). Fraction C was less for wheat (10.6%) than triticale (12.0%), CS (13.7%), and barley (16.4%). The ERD was greater for wheat (90.8%) compared with triticale (89.0%), CS (88.0%), and barley (85.6%). The kd for barley (2.4%) was less than triticale (4.9%). The kd for CS (1.8%) was less than triticale (4.9%). Overall, triticale had the greatest fraction A, barley had the greatest fraction B, and wheat had the least fraction C and the greatest ERD and kd. [ABSTRACT FROM AUTHOR]

The objective of the study was to evaluate the effects of partially replacing soybean meal (SBM) with black soldier fly larvae frass (BSFL) on nutrient digestibility, growth performance, and blood profile of ewes consuming diets with approximately 10% SBM on a DM basis. Six yearling Florida native ewes, averaging (mean ± SD) 44 ± 3 kg BW were enrolled in a replicated 3 x 3 Latin square design with 3 experimental periods of 21 d each (14 d adaptation + 4 d samples collection + 3 d washout). First 14 d of each period, ewes were housed in 7.4 x 1.8 m individual pens with rubber mats, one automatic waterer and two plastic feeders. During collection, ewes were moved to 1.4 x 0.7 m metabolic crates equipped with an individual waterer and feeder and two trays for the collection of feces and urine. Animals were fed TMR ad libitum twice daily at 0730 and 1530 h consisting of different levels of SBM replacement with BSFL, as follows: no replacement (CON), partial replacement of SBM with 25% BSFL (BSFL25) and 50% BSFL (BSFL50) on a DM basis. Data on DM and nutrient intake, excretion, and apparent digestibility, and also plasma concentrations of glucose and blood urea nitrogen (BUN), and urine concentrations of allantoin, urine blood nitrogen, and uric acid, were collected. Data were analyzed with SAS 9.4 evaluating orthogonal polynomial contrasts to determine linear and quadratic effects of SBM replacement with BSFL for each response variable. The intakes of DM, NDF, ADF, EE, and starch were greater for BSFL25 compared with control (Quadratic effect; P < 0.01) while no differences were observed between BSFL25 and BSFL50. Similarly apparent digestibilities of DM, NDF, ADF and starch was less for BSFL25 than CON (Quadratic effect; P ≤ 0.01). Intake of CP linearly increased (P < 0.01) with BSFL, while apparent digestibility of CP linearly decreased and that of EE increased, with inclusion of BSFL. There was a quadratic effect of BSFL on total feces excretion (P < 0.01) and water consumption (P < 0.01) being greater in BSFL25 compared with CON. Urine BUN (P < 0.01) and urine nitrogen (P < 0.01) linearly increased with BSFL inclusion. Results indicate that partial replacement of SBM with BSFL in the diet of yearling Florida native ewes may quadratically increase feed intake and decrease nutrient apparent digestibility. Further research is needed to explore the long-term implications of incorporating BSFL into animal diets, considering their impact on overall growth and performance. [ABSTRACT FROM AUTHOR]

The objectives of this study were: 1) to compare the effects of live yeast (LY), yeast fermentation product (YFP), a mix of Lactobacillus acidophilus and Propionibacterium freudenreichii (MLP), and Lactobacillus plantarum included as additives in dairy cows' diets on in vitro ruminal fermentation and gas production (GP); and 2) to evaluate the effects of L. plantarum as direct-fed microbials (DFM) in dairy cows' diets on in vitro ruminal fermentation, GP, nutrient digestibility, and N metabolism. Three experiments were carried out: Exp. 1 had the objective to compare all additives regarding ruminal fermentation parameters: an Ankom GP system was used in a completely randomized design, consisting of four 48 h incubations, and eight replications per treatment. There were eight treatments: a basal diet without additive (CTRL) or with one of the following additives: LY, YFP, MLP, or L. plantarum at four levels (% of diet Dry Matter (DM)): 0.05% (L1), 0.10% (L2), 0.15% (L3), and 0.20% (L4). In Exp. 2, a batch culture was used to evaluate ruminal fermentation, and CO2 and CH4 production using the same treatments and a similar experimental design, except for having 16 replications per treatment. Based on Exp. 1 and 2 results, Exp. 3 aimed at evaluating the effects of the L. plantarum on ruminal true nutrient digestibility and N utilization in order to evaluate the use of L. plantarum as DFM. The treatments CTRL, MLP, L1, and L2 were used in a replicated 4 × 4 Latin square design using a dual-flow continuous culture system. Data were analyzed using linear and nonlinear regression; treatment means were compared through contrasts, and L treatments in Exp. 1 and 2 were tested for linear, quadratic, and cubic effects. In Exp. 1, all treatments containing additives tended to reduce OM digestibility as well as reduced total volatile fatty acids (VFA) concentration and total GP. The YFP had greater OM digestibility than LY, and MLP treatment had greater total VFA concentration compared to L. plantarum treatments. In Exp. 2, additives reduced CO2 production, and there were no major differences in CH4. In Exp. 3, all additives reduced NH3-N concentration. In conclusion, pH and lactate concentration were not affected in all three experiments regardless of additive tested, suggesting that these additives may not improve ruminal fermentation by pH modulation; and L. plantarum may improve ruminal N metabolism when used as DFM in high-producing dairy cows' diets, mainly by reducing NH3-N concentration. [ABSTRACT FROM AUTHOR]

The objective of this study was to evaluate the effects of different sources of cobalt in a lactating dairy cow diet on in vitro ruminal fermentation. All diets provided the same amount of supplemental Co (1 mg per kg of diet DM) included in a premix with a targeted inclusion level of 2.5% of diet DM. Treatments were cobalt carbonate (CON), and three alternative Co sources; alternative source 1 (AS1), alternative source 2 (AS2), and alternative source 3 (AS3). Eight dual-flow continuous culture fermenters were used in a replicated 4 × 4 Latin square. Experimental periods were 10 d (7 d adaptation and 3 d sample collection). Composite samples of daily effluents were collected and analyzed for NH3-N, lactate, and VFA concentration as well as for nutrient digestibility. Samples of ruminal content were collected from each fermenter at 0, 1, 2, 4, 6, and 8 h after feeding to determine lactate, NH3-N, and pH kinetics. All data were analyzed using PROC MIXED of SAS and the repeated variable of time was included for pH, lactate, and NH3-N. There were neither treatment effects nor interactions between treatment and time for pH, NH3-N, and lactate concentrations, indicating a similar Co kinetics across treatments. With regards to lactate pool, we did not observe treatment effects for L-lactate; however, we observed treatment effects for D-lactate and total-lactate as seen in Table 1. Analyses of total and individual VFA, N utilization, and nutrient degradation are still undergoing, hence, it is possible that Co sources have effects on ruminal fermentation. [ABSTRACT FROM AUTHOR]

Our aim was to determine whether the method used to estimate truly digestible neutral detergent fiber (tdNDF) affects calculated concentrations of total digestible nutrients (TDN1x) and net energy of lactation (NEL3x) of canola meal (CM). Samples were collected from 12 CM processing plants in Canada over 4 yr (2011 to 2014, n = 47) and analyzed for dry matter (DM), crude protein (CP), ether extract (EE), ash, neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin (ADL), and neutral detergent insoluble CP (NDICP). Ruminal in situ incubation of CM samples was performed at 0, 24, 48, 96, and 288 h to determine NDF fractions (A, B, and C), effective ruminal NDF digestibility (ERNDFD), and indigestible NDF (iNDF) of CM. Three tdNDF-estimation methods were evaluated: 1) National Research Council (NRC) = 0.75 × (NDF - NDICP - ADL) × {1- [ADL/ (NDF - NDICP)]0.667}; 2) iNDF = 0.75 × (NDF - NDICP - NDF remaining after 288 h in situ); and 3) ERNDFD estimated from in situ NDF digestion kinetics. Resulting tdNDF values were used for calculation of TDN1x and NEL3x according to NRC (2001) equations. Data were analyzed with MIXED procedure of SAS 9.4 to determine the effect of processing plant on chemical composition, NDF degradation kinetics and NEL3x of CM. Effect of tdNDF estimation method on calculated TDN1x and NEL3x of CM was also evaluated. Model for analysis of processing plant included the fixed effect of plant and the random effect of year (plant) as replication, while analysis of tdNDF methods included the fixed effect of tdNDF estimation method and the random effects of processing plant and of year(plant) as replication. There was an effect of processing plant on DM (P = 0.03), CP (P < 0.01), EE (P < 0.01), and NDF (P < 0.01) of CM. Processing plant also had an effect on NDF fractions A (P < 0.01) and B (P = 0.02) but did not affect fraction C and ERNDFD. The tdNDF estimation method had an effect on tdNDF (P < 0.01), TDN1x (P < 0.01), and NEL3x (P < 0.01) of CM, yielding average NEL3x values of 1.72, 1.87, and 2.07 Mcal/kg for NRC, iNDF, and ERNDFD, respectively. Our results indicate that calculated energy concentration of CM according to NRC (2001) equations varies depending on the method used for estimation of tdNDF. Further research will be needed to determine the most accurate estimation method., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)