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We evaluated the effects of a Bacillus-based direct-fed microbial (DFM) on total in vitro gas production, dry matter (DM), neutral detergent fiber (NDF), and starch disappearance of different feedstuffs and total mixed rations (TMR) in three different experiments. In experiment 1, six single fiber-based feedstuffs were evaluated: alfalfa hay, buffalo grass, beet pulp, eragrostis hay, oat hay, and smutsvinger grass. Experimental treatments were control (with no probiotic inoculation; CON) or incubation of a probiotic mixture containing Bacillus licheniformis and B. subtilis (3.2 × 109 CFU/g; DFM). The calculation of DFM dose under in vitro conditions was based on the assumption of a rumen capacity of 70 liter and the dose of 3 g of the DFM mixture/head/d (9.6 × 109 CFU). Total in vitro gas production, DM, and NDF disappearance were evaluated at 24- and 48 h posttreatment incubation. Mean treatment effects were observed at 24- and 48 h gas production (P < 0.0001), as DFM incubation increased in vitro gas production by 5.0% and 6.5%, respectively. For nutrient digestibility, mean DM digestibility was increased at 48 h (P = 0.05), whereas mean NDF digestibility increased at both timepoints by incubating DFM in vitro (P ≤ 0.02). In experiment 2, nine commercial dairy TMR were collected and evaluated for the same variables and treatments described in experiment 1, with the additional analysis of starch digestibility at 7 h post in vitro incubation. The only difference was the concentration of the DFM included, being representative for a dosage of 8.8 × 109 CFU/head/d. In vitro gas production was increased only at 48 h due to DFM incubation (P = 0.05), whereas DM and NDF digestibility were improved at 24 and 48 h (P ≤ 0.02). No treatment effects were observed on in vitro starch digestibility (P = 0.31). In experiment 3, a combined analysis of DM and NDF digestibility was performed by using quality values (NDF and crude protein or CP) of 16 substrates. Regardless of CP and NDF levels of the substrates, DFM improved in vitro 24 and 48 h DM and NDF digestibility (P ≤ 0.03). In summary, incubating a Bacillus-based DFM (B. licheniformis and B. subtilis; BOVACILLUS) improved mean in vitro gas production, DM, and NDF digestibility of single feedstuffs and commercial dairy TMR, highlighting the potential of this combination of Bacillus spp. to improve nutrient utilization, mainly fiber.

We aimed to evaluate the effects of feeding super-conditioned corn at different temperatures on intake, growth performance, total-tract starch digestibility, rumen fermentation, blood metabolites, and feeding behavior of dairy calves. Thirty-six Holstein female dairy calves (40 ± 1.72 kg of body weight, ± SD) were randomly assigned to 1 of the following 3 treatments: (1) ground corn (control; CON; n = 12), (2) corn super-conditioned at 75°C (T-75; n = 12), and (3) corn super-conditioned at 95°C (T-95; n = 12). Three mash starter feeds with an identical nutritional composition were blended with 5% chopped alfalfa hay and fed to individually-housed calves from d 3 to 77 of their birth. All calves were fed 4 L/d of pasteurized whole milk twice daily since d 3 to 56, followed by 2 L/d of morning feeding from d 57 to 63 of age. Calves were weaned on d 63 and remained in the study until d 77. The T-75 and T-95 diets increased total-tract starch digestibility compared with the CON diet. Dry matter intake and weaning or final BW were not affected by treatments; however, average daily gain and feed efficiency increased in calves fed T-95 in the overall period. The T-95 diet increased withers height and tended to increase hip height compared with other diets, but feeding behavior did not change throughout the experimental period. Ruminal pH decreased in calves fed the T-95 diet compared with T-75 and CON diets. The molar proportion of ruminal propionate increased, whereas the acetate-to-propionate ratio tended to decrease in calves fed the T-95 compared with CON diet. Calves fed the T-95 diet had the highest blood glucose concentration, whereas a trend for increased insulin concentration was observed in calves fed T-95 compared with other diets. In conclusion, super-conditioning temperature of corn (T-95 vs. T-75 and CON) improved the average daily gain, feed efficiency, and skeletal growth, but did not influence dry matter intake during the first 77 d of age. Finally, the total-tract starch digestibility increased, whereas ruminal pH dropped during the postweaning period as super-conditioning temperature elevated.

Grazing management and N-fertilizer have been reported to improve tropical forage productivity and quality, however, their effect on methane emission of grazing animals remains uncertain. Therefore, this study aimed to assess the effects of increasing application rates of nitrogen (N) fertilization of Marandu palisadegrass under continuous stocking on intake, digestibility, nitrogen balance, and enteric methane emissions of Nellore growing bulls. We hypothesized that changes in the forage nutritive value caused by N fertilization of pastures combined with adequate grazing management (e.g., greater crude protein [CP] and digestibility) would lead to an increase in animal productivity (e.g., greater average daily gain [ADG] and gain per area), and then, to a decrease in methane emission intensity. Treatments consisted of different annual application rates of nitrogen fertilization: 0, 75, and 150 kg N/ha using ammonium nitrate (32% N) as the nitrogen source. The experimental design was completely randomized, with three treatments and four replications (12 paddocks). Intake, digestibility, N balance, and methane emissions were measured in eight animals per treatment. CP intake, digestibility and N balance increased linearly with the increase in N fertilization (P < 0.05). In addition, stocking rate (SR) and ADG linearly increased from 1.75 animal unit (AU = 450 kg)/ha and 0.62 kg/d (0 kg N/ha) to 3.75 AU/ha and 0.82 kg/d (150 kg N/ha), respectively. Individual methane emissions nor methane emission intensity were affected by treatment with an average of 164.7 g/d and 199.7 g/kg ADG (P > 0.05). Annual N fertilization with ammonium nitrate between 75 and 150 kg N/ha in palisadegrass pastures under continuous stocking enhances animal performance per unit area yet not affecting neither methane production nor intensity.

A meta-analysis of 158 peer-reviewed articles was conducted to examine effects of inoculation with Lactobacillus buchneri (LB)-based inoculants (LBB) that did or did not include homolactic or obligate heterolactic bacteria on silage fermentation and aerobic stability. A complementary meta-analysis of 12 articles examined LBB inoculation effects on dairy cow performance. Raw mean differences between inoculant and control treatment means weighted by inverse variance were compared with a hierarchical effects model that included robust variance estimation. Meta-regression and subgrouping analysis were used to identify effects of covariates including forage type, application rate (≤104, 105, 106, or ≥ 107 cfu/g as fed), bacteria type (LB vs. LB plus other bacteria), enzyme inclusion, ensiling duration, and silo type (laboratory or farm scale). Inoculation with LBB increased acetate (62%), 1, 2 propanediol (364%) and propionate (30%) concentration and aerobic stability (73.8%) and reduced lactate concentration (7.2%), yeast counts (7-fold) and mold counts (3-fold). Feeding inoculated silage did not affect milk yield, dry matter intake, and feed efficiency in lactating dairy cows. However, forage type, inoculant composition, and dose effects on silage quality measures were evident. Inoculation with LBB increased aerobic stability of all silages except tropical grasses. Adding obligate homolactic or facultative heterolactic bacteria to LB prevented the small increase in DM losses caused by LB alone. The 105 and 106 cfu/g rates were most effective at minimizing DM losses while aerobic stability was only increased with 105, 106, and ≥ 107 cfu/g rates. Inoculation with LBB increased acetate concentration, reduced yeast counts and improved aerobic stability but did not improve dairy cow performance.

This study aimed to evaluate the effects of feeding ground, steam-flaked, or super-conditioned corn on production performance, rumen fermentation, nutrient digestibility, and milk fatty acid (FA) profile of lactating dairy cows. Twenty-four lactating Holstein cows (130 ± 12 d in milk) in a completely randomized block design experiment were assigned to 1 of 3 treatments that contained 31% of one of the following corn types: (1) ground corn; (2) steam-flaked corn; and (3) super-conditioned corn. Actual milk yield was greater in the super-conditioned corn diet than in the steam-flaked and ground corn diets. Dry matter intake, 3.5% fat-corrected milk and energy-corrected milk remained unaffected by treatments; however, milk fat concentration decreased in the super-conditioned corn diet compared with the ground and steam-flaked corn diets. The molar proportion of ruminal acetate decreased in the super-conditioned corn diet compared with the ground and steam-flaked corn diets, whereas the molar proportion of propionate spiked in the super-conditioned corn diet. Ruminal pH dropped in cows fed super-conditioned corn compared with the other 2 diets. A similar pattern was observed for ruminal NH3-N and acetate-to-propionate ratio. Total-tract starch digestibility increased the most in the super-conditioned corn diet followed by the steam-flaked and ground corn diets (96.8, 95.1, and 92.5%, respectively). The neutral detergent fiber digestibility declined in cows fed the super-conditioned corn diet as opposed to other diets (~3.9%). The concentrations of 16:0 and mixed-FA in milk fat dropped in the super-conditioned corn-based diet compared with the ground corn diet. Milk trans-10 18:1 FA increased, whereas trans-11 18:1 FA decreased in cows fed the super-conditioned diet. We concluded that super-conditioned corn has the potential to increase milk yield and starch digestibility in lactating dairy cows; however, reduced milk fat output caused by altering ruminal pH and ruminal FA biohydrogenation pathways may not be desirable in certain markets. Future research is warranted to investigate how super-conditioned corn affects feed efficiency.

Pasture-fed beef cattle production has been often criticized as a source of greenhouse gas emissions. However, ruminants efficiently convert foods of low nutritional value to humans (e.g., pasture and by-products) into beef, a high-quality food. The present study aimed assessing the ratio between carbon emission per human-edible protein produced (HePp) of Nellore young bulls under three levels of beef cattle intensification systems: lenient, semi-intensive, and intensive. During backgrounding, all animals were raised in marandu palisadegrass pastures managed in continuous stocking with variable stocking rate and fertilized with 0, 75, and 150 kg N/ha, as ammonium nitrate (32% N) for lenient, semi-intensive, and intensive systems, respectively. During the finishing phase, bulls from lenient system remained in the pasture, while animals from semi-intensive and intensive systems were moved to a feedlot barn. The total carbon emission was represented by the total GHG emissions converted to CO2 equivalent (CO2e). Human-edible protein produced was calculated for each production phase and for the entire production system based on animal body protein minus the inedible fraction of empty body weight. Data were analyzed using PROC MIXED procedures of SAS. During the backgrounding phase, the ratio CO2e/kg HePp of semi-intensive did not differ from other systems; however, the intensive system presented the greatest ratio (P< 0.05). During the finishing phase, the ratio was greater for the more intensive systems (P< 0.05). For the entire period, the ratio was greater for the intensified systems (P< 0.05). Although the HePp of bulls from the intensified systems was greater than the lenient system (P< 0.05), the gain was insufficient to offset the CO2e production. Our results suggest that cattle ranching intensification acquired by improvements in grazing management is the most powerful tool to mitigate environmental impact of grazing systems. The authors acknowledge the São Paulo Research Foundation (FAPESP #2017/18750-7; #2018/26492-0).

Better understanding of N2O emissions from interactions between urine or feces, with different levels of nitrogen (N) fertilization on tropical pastures would be helpful in developing strategies to mitigate emissions and improve N2O inventories. The objective of this study was to evaluate N2O emissions from feces or urine samples collected from Nellore young bulls, isolated or in association with different levels of N fertilizer. Study was designed as completely randomized block design with five replications per treatment. Treatments were two levels of excreta (Feces or Urine) and three levels of N fertilizer (0, 75 and 150 kg N ha-1). Ammonium nitrate was used as source of N (32% of N). One liter of urine or 1.5 kg of feces was used per chamber with the respective N dose directly applied on excreta sample. Gas emissions were evaluated using the static chamber closed method and quantified using gas chromatography. Chambers were closed for 40 min and gas samples were collected at 0, 20 and 40 min. After treatment imposition, gas was sampled three times during week 1, twice during week 2, 3, and 4 and once during week 5, 6, 7 and 8, and subsequently every 2 weeks for two consecutive years. Type of excreta samples did not affect N2O emissions. Additionally, associative effects of N fertilizer onto feces or urine was similar. Cumulative N2O emissions linearly increased with increasing levels of N regardless of source, and ranged from 5.95, 32.1 to 74.7 mg of N2O/m2, for 0, 75 and 150 kg/ha/year of N. Similarly, emissions factor of N2O were linearly increased with increased N fertilizer (0.15, 0.36 and 0.73%, for excreta, excreta + 75 kg N and excreta + 150 kg N). Therefore, the interaction between N fertilizer and excreta on N2O emissions in tropical pastures was additive.

Grazing management and N- fertilizer have been reported to improve tropical forage productivity and quality, however, their effect on methane emission of grazing animals remains uncertain. Therefore, this study aimed to assess the effects of increasing application rates of nitrogen (N) fertilization of Marandu palisadegrass under continuous stocking on intake, digestibility, nitrogen balance, and enteric methane emissions of Nellore growing bulls. We hypothesized that changes in the forage nutritive value caused by N-fertilization of pastures combined with adequate grazing management (e.g., greater crude protein and digestibility) would lead to an increase in animal productivity (e.g., greater average daily gain, ADG, and gain per area), and then, to a decrease in methane emission intensity. Treatments consisted of different annual application rates of nitrogen fertilization: 0, 75, and 150 kg N/ha using ammonium nitrate (32% N) as the nitrogen source. The experimental design was completely randomized, with three treatments and four replications (12 paddocks). Intake, digestibility, N balance, and methane emissions were measured in eight animals per treatment. Crude protein (CP) intake, digestibility and N balance increased linearly with the increase in N fertilization (P0.05). In addition, stocking rate (SR) and average daily gain (ADG) linearly increased from 1.75 animal unit (AU = 450 kg)/ha and 0.62 kg/d (0 kg N/ha) to 3.75 AU/ha and 0.82 kg/d (150 kg N/ha), respectively. Individual methane emissions nor methane emission intensity were affected by treatment with an average of 164.7 g/d and 199.7 g/kg ADG (P0.05). Annual N fertilization with ammonium nitrate between 75 and 150 kg N/ha in palisadegrass pastures under continuous stocking enhances animal performance per unit area yet not affecting neither methane production nor intensity.

The purpose of this literature review is to evaluate current research into and understanding of whole-plant sorghum silage production and the effect of feeding whole-plant sorghum silage on lactation performance of dairy cows. Sorghum's drought tolerance, water efficiency, and low cost of production make it an intriguing crop in areas where whole-plant corn silage production may be limited. Currently, urban land encroachment and reduced water availability have increased social and economic pressures on farms to improve crop production efficiency. As these challenges become more prevalent, greater reliance on sorghum can be expected because of its ability to produce high dry matter yields while maintaining nutritive value, even under less-than-ideal growing conditions. Moreover, whole-plant sorghum silage provides both physically effective fiber and energy through fiber and grain fractions. Advancements in sorghum genetics and mechanical processing have the potential to alleviate common challenges associated with whole-plant sorghum silage supplementation, such as increased neutral detergent fiber and decreased neutral detergent fiber digestibility, starch concentration, and starch digestibility. These nutritive challenges must be overcome for whole-plant sorghum silage to be a viable alternative to whole-plant corn silage.

Objective Our objective was to evaluate the effects of ensiling on the corn silage processing score (CSPS) and fermentation and long-chain fatty acids (LCFA) profiles of unprocessed whole-plant corn. Materials and Methods Eleven corn hybrids were harvested and chopped separately (unprocessed) into approximately 16-mm-length particles and divided in 4 subsamples of approximately 600 g each. Two subsamples were randomly assigned to each treatment (0 or 120 d of ensiling). All samples were vacuum sealed in nylon-polyethylene standard barrier vacuum pouches. Representative samples from 0 and 120 d were immediately frozen and further analyzed for nutrients, CSPS, and fermentation and LCFA profiles. Results and Discussion Concentration of DM was unaffected (P > 0.10) by ensiling and averaged 36.2% as fed. Crude fat was 0.42 percentage units greater (P = 0.005) and pH levels were lower (P = 0.001) after 120 d, likely due to lactic and acetic acids concentrations that were 7.7 and 1.0 percentage units greater (P 0.10) by ensiling and averaged 31.2% of DM. The CSPS (28.8%) was low in the unprocessed material and was unaffected (P > 0.10) by ensiling. No effects of ensiling were observed on LCFA profile of major fatty acids including C16:0, C18:0, C18:1, C18:2, and C18:3 fatty acids (P > 0.10). Implications and Applications Ensiling did not affect CSPS and LCFA profile.

This study was conducted to examine the effects of clay (CL) and Saccharomyces cerevisiae fermentation product (SCFP) on the ruminal bacterial community of Holstein dairy cows challenged with aflatoxin B1 (AFB1). A second objective was to examine correlations between bacterial abundance and performance measures. Eight lactating dairy cows stratified by milk yield and parity were randomly assigned to 4 treatments in a 4 × 4 Latin square design with 2 replicate squares, four 33-d periods, and a 5-d washout between periods. The treatments included (1) control (basal diet, no additive); (2) T (control + 63.4 µg/kg AFB1, oral dose); (3) CL (T + 200 g/head per day of sodium bentonite clay, top-dress); and (4) CL+SCFP [CL + 19 g/head per day Diamond V NutriTek (Diamond V Inc., Cedar Rapids, IA) + 16 g/head per day MetaShield (Diamond V Inc.), top-dress]. Cows were adapted to diets containing no AFB1 from d 1 to 25 (predosing period). From d 26 to 30 (dosing period), AFB1 was orally dosed and then withdrawn for d 31 to 33 (withdrawal period). During the predosing period, compared with the control, feeding CL and CL+SCFP increased the relative abundance of the most dominant phylum, Bacteroidetes (55.1 and 55.8 vs. 50.6%, respectively), and feeding CL+SCFP increased Prevotella abundance (43.3 and 43.6 vs. 40.0%, respectively). During the dosing period, feeding AFB1 did not affect the ruminal bacterial community, but the relative abundance of Fibrobacteraceae increased with CL+SCFP compared with T (1.45 vs. 0.97%); Fibrobacter abundance also tended to increase with CL+SCFP compared with T and control, respectively (1.45 vs. 0.97 and 1.05%, respectively). Feeding AFB1 with or without CL or CL+SCFP did not affect ruminal pH or concentrations of NH3-N, total volatile fatty acids, or individual volatile fatty acids. Milk yield and milk component yields were positively correlated with the relative abundance of unclassified Succinivibrionaceae, unclassified YS2, or Coprococcus. Feed efficiency was positively correlated (r ≥ 0.30) with the relative abundance of unclassified YS2, Coprococcus, or Treponema. Feeding aflatoxin at 63 µg/kg, a common contamination level on farms, did not affect the abundance of dominant bacteria or rumen fermentation. When aflatoxin was fed, CL+SCFP increased the abundance of Fibrobacter, a major fibrolytic bacteria genus. Milk yield and DMI were positively correlated with abundance of Succinivibrionaceae and Coprococcus. Feed efficiency was positively correlated with abundance of Coprococcus, Treponema, and YS2. Future studies should speciate culture and determine the functions of the bacteria to elucidate their roles in the rumen and potential contribution to increasing the performance of dairy cows.

Aflatoxin is a potent carcinogen commonly found in animal feeds that can impair rumen fermentation at high concentrations; however, its effects at physiologically relevant concentrations are unknown. This study examined the effects of aflatoxin B1 (AFB1), with or without bentonite clay (CL) and Saccharomyces cerevisiae fermentation product (SCFP)-based sequestering agents on in vitro rumen fermentation and digestibility of a dairy cow TMR. Corn silage-based TMR (0.5 g, 17.3% crude protein and 1.67 Mcal/kg of net energy for lactation) was incubated in a rumen fluid-buffer inoculum (1:2 ratio; 50 mL) with the following treatments: (1) no additives (control); (2) control + 0.75 µg/L AFB1 (T); (3) T + 80 mg/L sodium bentonite clay (CL; Astra-Ben-20, Prince Agri Products Inc., Quincy, IL); or (4) CL + 14 mg/L SCFP (CL+SCFP; Diamond V, Cedar Rapids, IA). Ruminal fluid was collected 3 h after the morning feeding from 3 cannulated cows fed the same TMR, and rumen fluid from individual cows was used to prepare separate inocula. Each treatment was incubated in duplicate at 39°C for 0, 4, 8, 16, and 24 h in each of 3 runs. Adding T reduced total volatile fatty acid (VFA) concentration after 4 and 8 h and molar proportion of propionate after 4 and 24 h of incubation relative to control. Adding sequestering agents (CL and CL+SCFP) with T did not affect total VFA concentration after 4 or 8 h, but increased total VFA after 16 h and tended to increase molar proportion of propionate after 24 h compared with T. At 24 h, T had lower DM digestibility and higher NH3-N concentration compared with the control. Thus, AFB1, even at very low concentration (0.75 µg/L), had detrimental effects on rumen fermentation and subsequently DM digestibility of the TMR. Adding sequestering agents did not prevent negative effects of T on rumen fermentation within 8 h of incubation; however, sequestering agents were effective after 16 h of incubation.

This study examined the effects of chopping or chopping + blender maceration of red clover or lucerne on pre-fermented juice (PFJ) and determined the effects of PFJs on the quality of red clover silage or lucerne silage. The PFJs from chopping red clover (PFJ-RC) or lucerne (PFJ-LC) had a higher lactic acid bacteria (LAB) count than that from chopping + blender maceration (p &lt, 0.05) and were used as additives. Compared with the Control of both silages, adding PFJ increased LAB, lactic acid (LA), and in vitro digestibility of dry matter (IVDMD) (p &lt, 0.05), while pH, acetic acid (AA), and ammonia nitrogen/total nitrogen (NH3-N/TN) were decreased (p &lt, 0.05). For red clover silages, the PFJ-RC treatment contained the greatest LAB and LA and the lowest pH and NH3-N/TN among treatments (p &lt, 0.05), similar results were observed in PFJ-LC treatment for lucerne silages (p &lt, 0.05). The IVDMD of both silages correlated negatively with pH, AA, and NH3-N/TN and positively with LA (p &lt, 0.05). Overall, chopping alone was a better method for preparing PFJ. Adding PFJ at ensiling increased LA and decreased the pH, AA, and NH3-N/TN of both silages. Ensiling lucerne or red clover with PFJ from the ensiling material had a more positive effect on the fermentation parameters mentioned above. Satisfactory fermentation parameters detected in the present study contributed to improving the IVDMD of both silages.

Four experiments were conducted to examine the effects of a recombinant bacterial expansin-like protein (BsEXLX1) from Bacillus subtilis and a commercial exogenous fibrolytic enzyme (EFE) preparation for ruminants on hydrolysis of pure substrates (cellulose and xylan) and in vitro digestibility of bermudagrass haylage (BMH). Recombinant Escherichia coli BL21 strain was used to express BsEXLX1; the protein was purified using an affinity column. In experiment 1, carboxymethylcellulose, Whatman #1 filter paper (General Electric, Boston, MA) and oat-spelt xylan substrates were subjected to 4 treatments (1) sodium citrate buffer (control), (2) BsEXLX1 (162 µg/g of substrate), (3) EFE (2.3 mg/g of substrate), and (4) EFE + BsELX1 in 3 independent runs. Samples were incubated at optimal conditions for both additives (pH 5 and 50°C) or at ruminal (pH 6 and 39°C) or ambient (pH 6 and 25°C) conditions for 24 h and sugar release was measured. In experiment 2, digestibility in vitro of BMH was examined after treatment with the following: (1) control (buffer only), (2) BsEXLX1 (162 µg/g of dry matter), (3) EFE (2.2 mg/g of dry matter), and (4) EFE + BsEXLX1 in 3 independent runs at 39°C for 24 h. Experiment 3 examined effects of EFE and BsEXLX1 on simulated preingestive hydrolysis and profile of released sugars from BMH after samples were suspended in deionized water with sodium azide at 25°C for 24 h in 2 independent runs. In experiment 4, the sequence of the BsEXLX1 purified protein was compared with 447 ruminal bacterial genomes to identify similar proteins from the rumen. In experiment 1, compared with EFE alone, EFE and BsEXLX1 synergistically increased sugar release from carboxymethylcellulose and Whatman #1 filter paper under all simulated conditions; however, hydrolysis of xylan was not improved. In experiment 2, compared with EFE alone, treatment with EFE and BsEXLX1 increased neutral detergent fiber and acid detergent fiber digestibility of bermudagrass haylage (by 5.5 and 15%, respectively) and total volatile fatty acid concentrations, and decreased acetate-propionate ratio. In experiment 3, compared with EFE alone. The EFE and BsEXLX1 synergistically reduced concentrations of neutral detergent fiber and acid detergent fiber and increased release of sugars by 9.3%, particularly cellobiose (72.5%). In experiment 4, a similar sequence to that of BsEXLX1 was identified in Bacillus licheniformis, and similar hypothetical protein sequences were identified in Ruminococcus flavefaciens strains along with different protein structures in E. xylanophilum and Lachnospiraceae. This study showed that an expansin-like protein synergistically increased the hydrolysis of pure cellulose substrates and the hydrolysis and digestibility in vitro of BMH.

Background The major greenhouse gas from ruminants is enteric methane (CH4) which in 2010, was estimated at 2.1 Gt of CO2 equivalent, accounting for 4.3% of global anthropogenic greenhouse gas emissions. There are extensive efforts being made around the world to develop CH4 mitigating inhibitors that specifically target rumen methanogens with the ultimate goal of reducing the environmental footprint of ruminant livestock production. This study examined the individual and combined effects of supplementing a high-forage diet (90% barley silage) fed to beef cattle with the investigational CH4 inhibitor 3-nitrooxypropanol (3-NOP) and canola oil (OIL) on the rumen microbial community in relation to enteric CH4 emissions and ruminal fermentation. Results 3-NOP and OIL individually reduced enteric CH4 yield (g/kg dry matter intake) by 28.2% and 24.0%, respectively, and the effects were additive when used in combination (51.3% reduction). 3-NOP increased H2 emissions 37-fold, while co-administering 3-NOP and OIL increased H2 in the rumen 20-fold relative to the control diet. The inclusion of 3-NOP or OIL significantly reduced the diversity of the rumen microbiome. 3-NOP resulted in targeted changes in the microbiome decreasing the relative abundance of Methanobrevibacter and increasing the relative abundance of Bacteroidetes. The inclusion of OIL resulted in substantial changes to the microbial community that were associated with changes in ruminal volatile fatty acid concentration and gas production. OIL significantly reduced the abundance of protozoa and fiber-degrading microbes in the rumen but it did not selectively alter the abundance of rumen methanogens. Conclusions Our data provide a mechanistic understanding of CH4 inhibition by 3-NOP and OIL when offered alone and in combination to cattle fed a high forage diet. 3-NOP specifically targeted rumen methanogens and partly inhibited the hydrogenotrophic methanogenesis pathway, which increased H2 emissions and propionate molar proportion in rumen fluid. In contrast, OIL caused substantial changes in the rumen microbial community by indiscriminately altering the abundance of a range of rumen microbes, reducing the abundance of fibrolytic bacteria and protozoa, resulting in altered rumen fermentation. Importantly, our data suggest that co-administering CH4 inhibitors with distinct mechanisms of action can both enhance CH4 inhibition and provide alternative sinks to prevent excessive accumulation of ruminal H2.

Aflatoxins are poisonous carcinogens produced by fungi, mainly Aspergillus flavus and Aspergillus parasiticus. Aflatoxins can contaminate a variety of livestock feeds and cause enormous economic losses, estimated at between US$52.1 and US$1.68 billion annually for the U.S. corn industry alone. In addition, aflatoxin can be transferred from the diet to the milk of cows as aflatoxin M1 (AFM1), posing a significant human health hazard. In dairy cows, sheep and goats, chronic exposure to dietary aflatoxin can reduce milk production, impair reproduction and liver function, compromise immune function, and increase susceptibility to diseases; hence, strategies to lower aflatoxin contamination of feeds and to prevent or reduce the transfer of the toxin to milk are required for safeguarding animal and human health and improving the safety of dairy products and profitability of the dairy industry. This article provides an overview of the toxicity of aflatoxin to ruminant livestock, its occurrence in livestock feeds, and the effectiveness of different strategies for preventing and mitigating aflatoxin contamination of feeds.

This 7-page document provides a list of common hazards to consider in the manufacturing of feed for small ruminants. Written by Chalise Brown, Diwakar Vyas, and Jason M. Scheffler, and published by the UF/IFAS Department of Animal Sciences, November 2020.

The individual and combined effects of 3-nitrooxypropanol (3-NOP) and canola oil (OIL) supplementation on enteric methane (CH4) and hydrogen (H2) emissions, rumen fermentation and biohydrogenation, and total tract nutrient digestibility were investigated in beef cattle. Eight beef heifers (mean body weight ± SD, 732 ± 43 kg) with ruminal fistulas were used in a replicated 4 × 4 Latin square with a 2 (with and without 3-NOP) × 2 (with and without OIL) arrangement of treatments and 28-d periods (13 d adaption and 15 d measurements). The four treatments were: control (no 3-NOP, no OIL), 3-NOP (200 mg/kg dry matter [DM]), OIL (50 g/kg DM), and 3-NOP (200 mg/kg DM) plus OIL (50 g/kg DM). Animals were fed restrictively (7.6 kg DM/d) a basal diet of 900 g/kg DM barley silage and 100 g/kg DM supplement. 3-NOP and OIL decreased (P < 0.01) CH4 yield (g/kg DM intake) by 31.6% and 27.4%, respectively, with no 3-NOP × OIL interaction (P = 0.85). Feeding 3-NOP plus OIL decreased CH4 yield by 51% compared with control. There was a 3-NOP × OIL interaction (P = 0.02) for H2 yield (g/kg DM intake); the increase in H2 yield (P < 0.01) due to 3-NOP was less when it was combined with OIL. There were 3-NOP × OIL interactions for molar percentages of acetate and propionate (P < 0.01); individually, 3-NOP and OIL decreased acetate and increased propionate percentages with no further effect when supplemented together. 3-NOP slightly increased crude protein (P = 0.02) and starch (P = 0.01) digestibilities, while OIL decreased the digestibilities of DM (P < 0.01) and neutral detergent fiber (P < 0.01) with no interactions (P = 0.15 and 0.10, respectively). 3-NOP and OIL increased (P = 0.04 and P < 0.01, respectively) saturated fatty acid concentration in rumen fluid, with no interaction effect. Interactions for ruminal trans-monounsaturated fatty acids (t-MUFA) concentration and percentage were observed (P = 0.02 and P < 0.01); 3-NOP had no effect on t-MUFA concentration and percentage, while OIL increased the concentration (P < 0.01) and percentage (P < 0.01) of t-MUFA but to a lesser extent when combined with 3-NOP. In conclusion, the CH4-mitigating effects of 3-NOP and OIL were independent and incremental. Supplementing ruminant diets with a combination of 3-NOP and OIL may help mitigate CH4 emissions, but the decrease in total tract digestibility due to OIL may decrease animal performance and needs further investigation.

Two separate experiments were carried out to evaluate the effects of incremental doses of 10 exogenous endo-acting α-amylase and exo-acting glucoamylase; 1LAT (bacterial α-amylase), 2AK, 3AC, 4Cs4, 5Trga, 6Afuga, 7Fvga, and 10Tg (fungal α-amylases, glucoamylases, and α-glucosidase), 8Star and 9Syn (fungal amylase-mixtures; experiment 1) and three exogenous proteases; 11P14L, 12P7L, and 13P30L (bacterial proteases; experiment 2) on in vitro dry matter digestibility (IVDMD) and in vitro starch digestibility (IVSD) of mature dent corn grain using a batch culture system. Incremental doses of the exogenous enzymes (0, 0.25, 0.50, 0.75, and 1.00 mg/g of dried substrate) were applied directly to the substrate (0.5 g of ground corn, 4 mm) in sextuplicate (experiment 1) or quadruplicate (experiment 2) within F57 filter bags, which were incubated at 39 °C in buffered rumen fluid for 7 h. Rumen fluid was collected 2–3 h after the morning feeding from three lactating dairy cows and pooled. Cows were consuming a midlactation total mixed ration (TMR; 1.60 Mcal/kg DM and 15.4%; net energy of lactation and crude protein, respectively). Three independent runs were carried out for each experiment. Data were analyzed as a randomized complete block design using run as the blocking factor. Dose was used as a fixed factor while run was considered a random factor. Linear, quadratic, and cubic orthogonal contrasts were also tested. In experiment 1, enzymes 2AK, 3AC, and 10Tg did not increase (P > 0.10) IVDMD and IVSD, whereas 0.25 mg of enzymes 1LAT, 5Trga, and 8Star increased (P < 0.01) IVDMD by 23%, 47%, and 62% and IVSD by 35%, 41%, and 58%, respectively, compared with the control. Enzymes 4Cs4, 6Afuga, 7Fvga, and 9Syn linearly increased IVDMD and IVSD (P < 0.01). Greatest increases in IVDMD (82.9%) and IVSD (85.9%) resulted with 1 mg of 6Afuga compared to control. In experiment 2, the lowest dose of exogenous proteases 11P14L and 12P7L increased (P < 0.01) IVDMD by 98% and 87% and IVSD by 57% and 64%, respectively, whereas the highest dose of 13P30L increased (P = 0.02) IVDMD by 44.8% and IVSD by 30%, relative to the control. In conclusion, IVSD and IVDMD were increased by one α-amylase, certain glucoamylases, and all proteases tested, with the glucoamylase 6Afuga in experiment 1 and the neutral protease 12P7L in experiment 2, increasing IVDMD and IVSD to the greater extents. Future in vivo studies are required to validate these findings before these enzyme additives can be recommended for improving the digestibility of mature dent corn grain.

We previously reported that milk production in dairy cows was increased by adding a specific xylanase-rich exogenous fibrolytic enzyme (XYL) to a total mixed ration (TMR) containing 10% bermudagrass silage (BMD). Two follow-up experiments were conducted to examine whether adding XYL would increase the performance of dairy cows consuming a TMR containing a higher (20%) proportion of BMD (Experiment 1) and to evaluate the effects of XYL on in vitro fermentation and degradability of the corn silage, BMD, and TMR (Experiment 2). In Experiment 1, 40 lactating Holstein cows in early lactation (16 multiparous and 24 primiparous; 21 ± 3 d in milk; 589 ± 73 kg of body weight) were blocked by milk yield and parity and randomly assigned to the Control and XYL treatments. The TMR contained 20% BMD, 25% corn silage, 8% wet brewer's grain, and 47% concentrate mixture in the dry matter (DM). Cows were fed the XYL-treated or untreated experimental TMR twice per day for 10 wk after a 9-d covariate period. In Experiment 2, ruminal fluid was collected from 3 cannulated lactating Holstein cows fed a diet containing 20% bermudagrass haylage, 25% corn silage and 55% concentrate. In Experiment 1, compared with Control, application of XYL did not affect DM intake (24.0 vs. 23.7 kg/d), milk yield (35.1 vs. 36.2 kg/d), fat-corrected milk yield (36.1 vs. 36.9 kg/d), or yields of milk fat (1.29 vs. 1.31 kg/d) or protein (1.07 vs. 1.08 kg/d). However, intake of neutral detergent fiber (4.67 vs. 4.41 kg/d) tended to increase with XYL; consequently, milk protein concentration was increased by XYL (3.02 vs. 2.95%). Feed efficiency tended to be lower in cows fed XYL (1.57 vs. 1.52 kg of fat-corrected milk/kg of DM intake) compared with Control. In Experiment 2, XYL tended to increase the rate of gas production in the TMR, the molar proportion of propionate for corn silage, and that of valerate for the TMR. In addition, XYL increased in vitro DM, neutral detergent fiber, and acid detergent fiber degradability of BMD and corn silage. Application of XYL to a diet with a relatively high proportion of BMD tended to increase digestible neutral detergent fiber intake, increased milk protein concentration, and in vitro degradability of DM, neutral detergent fiber, and acid detergent fiber. However, XYL did not affect milk production and tended to decrease feed efficiency in early lactation cows.

Dairy is the most important subsector in the Sri Lankan livestock industry, due to the need to address the growing demand for fresh milk and milk products, and because of its potential influence on the rural economy. The USDA Food for Progress program awarded a 4.5-year Market-Oriented Dairy project to International Executive Service Corps, a not-for-profit organization based in Washington, DC. The objective of the Market-Oriented Dairy project is to support Sri Lanka's dairy sector and catalyze sustainable growth by strengthening the dairy sector through better technological, financial, and management practices benefiting all stakeholders and consumers along the dairy value chain. The University of Florida is working with International Executive Service Corps as technical experts in conducting dairy value chain assessments, identifying gaps and challenges in dairy management practices, extension services, milk quality management standards, and artificial insemination services. Assessment of the dairy value chain in 2018 identified a lack of good quality and quantity of feed, along with poor dairy management practices and ineffective extension services as major constraints to improving dairy productivity in Sri Lanka. In addition, lack of national milk quality standards that are consistent with international benchmarks and inadequate cooling facilities are significant challenges to improving milk quality. The nutritional status of cows is not suitable for optimal reproductive performance, compromising the success of artificial insemination in Sri Lanka. Based on these findings, we developed a dairy assessment tool and provided comprehensive training sessions targeting extension agents, veterinarians, and farmers to promote best practices in dairy management. Beyond training, however, industry support for standardization and monitoring of milk and feed quality are needed, providing opportunities for private investment to support the dairy industry. Similar opportunities are available for forage production and delivery to producers. The broader aim of the Market-Oriented Dairy project intervention is to reduce Sri Lanka's dependency on imported milk and contribute toward the goal of a safe, self-sufficient fresh milk supply.

Undesirable interactions between trace mineral elements and ruminal contents may occur during digestion when mineral salts are supplemented. Antimicrobial effects of copper sulfate (CuSO4) may affect ruminal digestibility of nutrients when fed as a source of copper (Cu), while sodium selenite (Na2SeO3) may be reduced in the rumen to less available forms of selenium (Se). Our objective was to evaluate if protection of CuSO4 and Na2SeO3 by lipid-microencapsulation would induce changes on ruminal microbial fermentation. We used 8 fermentors in a dual-flow continuous-culture system in a 4 × 4 duplicated Latin square with a 2 × 2 factorial arrangement of treatments. Factors were CuSO4 protection (unprotected and protected by lipid-microencapsulation) and Na2SeO3 protection (unprotected and protected by lipid-microencapsulation). Treatments consisted of supplementation with 15 mg/kg of Cu and 0.3 mg/kg of Se from either unprotected or protected (lipid-microencapsulated) sources, as follows: (1) Control (unprotected CuSO4 + unprotected Na2SeO3); (2) Cu-P (protected CuSO4 + unprotected Na2SeO3); (3) Se-P (unprotected CuSO4 + protected Na2SeO3); (4) (Cu+Se)-P (protected CuSO4 + protected Na2SeO3). All diets had the same nutrient composition and fermentors were fed 106 g of dry matter/d. Each experimental period was 10 d (7 d of adaptation and 3 d for sample collections). Daily pooled samples of effluents were analyzed for pH, NH3-N, nutrient digestibility, and flows (g/d) of total N, NH3-N, nonammonia N (NAN), bacterial N, dietary N, and bacterial efficiency. Kinetics of volatile fatty acids was analyzed in samples collected daily at 0, 1, 2, 4, 6, and 8 h after feeding. Main effects of Cu protection, Se protection, and their interaction were tested for all response variables. Kinetics data were analyzed as repeated measures. Protection of Cu decreased acetate molar proportion, increased butyrate proportion, and tended to decrease acetate:propionate ratio in samples of kinetics, but did not modify nutrient digestibility. Protection of Se tended to decrease NH3-N concentration, NH3-N flow, and CP digestibility; and to increase flows of nonammonia N and dietary N. Our results indicate that protection of CuSO4 may increase butyrate concentration at expenses of acetate, while protection of Na2SeO3 tended to reduce ruminal degradation of N. Further research is needed to determine the effects of lipid-microencapsulation on intestinal absorption, tissue distribution of Cu and Se, and animal performance.

The study objective was to evaluate the combined effects of supplementing monensin (MON) and the methane (CH4) inhibitor 3-nitrooxypropanol (NOP) on enteric CH4 emissions, growth rate, and feed conversion efficiency of backgrounding and finishing beef cattle. Two hundred and forty crossbred steers were used in a 238-d feeding study and fed a backgrounding diet for the first 105 d (backgrounding phase), transition diets for 28 d, followed by a finishing diet for 105 d (finishing phase). Treatments were as follows: 1) control (no additive); 2) MON (monensin supplemented at 33 mg/kg DM; 3) NOP (3-nitrooxypropanol supplemented at 200 mg/kg DM for backgrounding or 125 mg/kg DM for finishing phase); and 4) MONOP (33 mg/kg DM MON supplemented with either 200 mg/kg DM or 125 mg/kg DM NOP). The experiment was a randomized complete block (weight: heavy and light) design with 2 (NOP) × 2 (MON) factorial arrangement of treatments using 24 pens (8 cattle/pen; 6 pens/treatment) at the main feedlot and 8 pens (6 cattle/pen; 2 pens/treatment) at the controlled environment building (CEB) feedlot. Five animals per treatment were moved to chambers for CH4 measurements during both phases. Data were analyzed using a Mixed procedure of SAS with pen as experimental unit (except CH4). Location (Main vs. CEB) had no significant effect and was thus omitted from the final model. Overall, there were few interactions between MON and NOP indicating that the effects of the 2 compounds were independent. When cattle were fed the backgrounding diet, pen DMI was decreased by 7%, whereas gain-to-feed ratio (G:F) was improved by 5% with NOP supplementation (P < 0.01). Similarly, MON improved G:F ratio by 4% (P < 0.01), but without affecting DMI. During the finishing phase, DMI tended (P = 0.06) to decrease by 5% with both MON (5%) and NOP (5%), whereas ADG tended (P = 0.08) to decrease by 3% with MON. Gain-to-feed ratio for finishing cattle was improved with NOP by 3% (P < 0.01); however, no effects were observed with MON. 3-Nitrooxypropanol decreased CH4 yield (g/kg DMI) by 42% and 37% with backgrounding and finishing diets (P ≤ 0.01), respectively, whereas MON did not lower CH4 yield. Overall, these results demonstrate efficacy of NOP in reducing enteric CH4 emissions and subsequently improving feed conversion efficiency in cattle fed high-forage and high-grain diets. Furthermore, effects of NOP did not depend on whether MON was included in the diet.

Ensiled forage, particularly corn silage, is an important component of dairy cow diets worldwide. Forages can be contaminated with several mycotoxins in the field pre-harvest, during storage, or after ensiling during feed-out. Exposure to dietary mycotoxins adversely affects the performance and health of livestock and can compromise human health. Several studies and surveys indicate that ruminants are often exposed to mycotoxins such as aflatoxins, trichothecenes, ochratoxin A, fumonisins, zearalenone, and many other fungal secondary metabolites, via the silage they ingest. Problems associated with mycotoxins in silage can be minimized by preventing fungal growth before and after ensiling. Proper silage management is essential to reduce mycotoxin contamination of dairy cow feeds, and certain mold-inhibiting chemical additives or microbial inoculants can also reduce the contamination levels. Several sequestering agents also can be added to diets to reduce mycotoxin levels, but their efficacy varies with the type and level of mycotoxin contamination. This article gives an overview of the types, prevalence, and levels of mycotoxin contamination in ensiled forages in different countries, and describes their adverse effects on health of ruminants, and effective prevention and mitigation strategies for dairy cow diets. Future research priorities discussed include research efforts to develop silage additives or rumen microbial innocula that degrade mycotoxins.

The study was conducted to examine the effect of supplementing bentonite clay with or without a Saccharomyces cerevisiae fermentation product (SCFP; 19 g of NutriTek + 16 g of MetaShield, both from Diamond V, Cedar Rapids, IA) on the performance and health of dairy cows challenged with aflatoxin B1 (AFB1). Twenty-four lactating Holstein cows (64 ± 11 d in milk) were stratified by parity and milk production and randomly assigned to 1 of 4 treatment sequences. The experiment had a balanced 4 × 4 Latin square design with 6 replicate squares, four 33-d periods, and a 5-d washout interval between periods. Cows were fed a total mixed ration containing 36.1% corn silage, 8.3% alfalfa hay, and 55.6% concentrate (dry matter basis). Treatments were (1) control (no additives), (2) toxin (T; 1,725 µg of AFB1/head per day), (3) T + clay (CL; 200 g/head per day; top-dressed), and (4) CL+SCFP (CL+SCFP; 35 g/head per day; top-dressed). Cows were adapted to diets from d 1 to 25 (predosing period) and then orally dosed with AFB1 from d 26 to 30 (dosing period), and AFB1 was withdrawn from d 31 to 33 (withdrawal period). Milk samples were collected twice daily from d 21 to 33, and plasma was sampled on d 25 and 30 before the morning feeding. Transfer of ingested AFB1 into milk aflatoxin M1 (AFM1) was greater in T than in CL or CL+SCFP (1.65 vs. 1.01 and 0.94%, respectively) from d 26 to 30. The CL and CL+SCFP treatments reduced milk AFM1 concentration compared with T (0.45 and 0.40 vs. 0.75 µg/kg, respectively), and, unlike T, both CL and CL+SCFP lowered AFM1 concentrations below the US Food and Drug Administration action level (0.5 µg/kg). Milk yield tended to be greater during the dosing period in cows fed CL+SCFP compared with T (39.7 vs. 37.7 kg/d). Compared with that for T, plasma glutamic oxaloacetic transaminase concentration, indicative of aflatoxicosis and liver damage, was reduced by CL (85.9 vs. 95.2 U/L) and numerically reduced by CL+SCFP (87.9 vs. 95.2 U/L). Dietary CL and CL+SCFP reduced transfer of dietary AFB1 to milk and milk AFM1 concentration. Only CL prevented the increase in glutamic oxaloacetic transaminase concentration, and only CL+SCFP prevented the decrease in milk yield caused by AFB1 ingestion.

The first objective of this study was to examine effects of adding Escherichia coli O157:H7 with or without chemical or microbial additives on the bacterial diversity and composition of alfalfa silage. The second objective was to examine associations between the relative abundance of known and unknown bacterial species and indices of silage fermentation quality. Alfalfa forage was harvested at 54% dry matter, chopped to a theoretical length of cut of 19 mm, and ensiled in quadruplicate in laboratory silos for 100 d after the following treatments were applied: (1) distilled water (control); (2) 1 × 10 5 cfu/g of E. coli O157:H7 (EC); (3) EC and 1 × 10 6 cfu/g of Lactobacillus plantarum (EC+LP); (4) EC and 1 × 10 6 cfu/g of Lactobacillus buchneri (EC+LB); and (5) EC and 0.22% propionic acid (EC+PA). After 100 d of ensiling, the silage samples were analyzed for bacterial diversity and composition via the Illumina MiSeq platform (Illumina Inc., San Diego, CA) and chemically characterized. Overall, Firmicutes (74.1 ± 4.86%) was the most predominant phylum followed by Proteobacteria (20.4 ± 3.80%). Relative to the control, adding E. coli O157:H7 alone at ensiling did not affect bacterial diversity or composition but adding EC+LP or EC+LB reduced the Shannon index, a measure of diversity (3.21 vs. 2.63 or 2.80, respectively). The relative abundance of Firmicutes (69.2 and 68.8%) was reduced, whereas that of Proteobacteria (24.0 and 24.9%) was increased by EC+LP and EC+PA treatments, relative to those of the control (79.5 and 16.5%) and EC+LB (77.4 and 18.5%) silages, respectively. Compared with the control, treatment with EC+LP increased the relative abundance of Lactobacillus, Sphingomonas, Pantoea, Pseudomonas , and Erwinia by 426, 157, 200, 194, and 163%, respectively, but reduced those of Pediococcus, Weissella , and Methylobacterium by 5,436, 763, and 250%, respectively. Relative abundance of Weissella (9.19%) and Methylobacterium (0.94%) were also reduced in the EC+LB silage compared with the control (29.7 and 1.50%, respectively). Application of propionic acid did not affect the relative abundance of Lactobacillus, Weissella , or Pediococcus. Lactate concentration correlated positively (r = 0.56) with relative abundance of Lactobacillus and negatively (r = −0.41) with relative abundance of Pediococcus . Negative correlations were detected between ammonia-N concentration and relative abundance of Sphingomonas (r = −0.51), Pantoea (r = −0.46), Pseudomonas (r = −0.45), and Stenotrophomonas (r = −0.38). Silage pH was negatively correlated with relative abundance of Lactobacillus (r = −0.59), Sphingomonas (r = −0.66), Pantoea (r = −0.69), Pseudomonas (r = −0.69), and Stenotrophomonas (r = −0.50). Future studies should aim to speciate, culture, and determine the functions of the unknown bacteria detected in this study to elucidate their roles in silage fermentation.

The objectives of this study were to evaluate the relationship between residual feed intake (RFI; g/d) and enteric methane (CH) production (g/kg DM) and to compare CH and carbon dioxide (CO) emissions measured using respiration chambers (RC) and the GreenFeed emission monitoring (GEM) system (C-Lock Inc., Rapid City, SD). A total of 98 crossbred replacement heifers were group housed in 2 pens and fed barley silage ad libitum and their individual feed intakes were recorded by 16 automated feeding bunks (GrowSafe, Airdrie, AB, Canada) for a period of 72 d to determine their phenotypic RFI. Heifers were ranked on the basis of phenotypic RFI, and 16 heifers (8 with low RFI and 8 with high RFI) were randomly selected for enteric CH and CO emissions measurement. Enteric CH and CO emissions of individual animals were measured over two 25-d periods using RC (2 d/period) and GEM systems (all days when not in chambers). During gas measurements metabolic BW tended to be greater ( ≤ 0.09) for high-RFI heifers but ADG tended ( = 0.09) to be greater for low-RFI heifers. As expected, high-RFI heifers consumed 6.9% more feed ( = 0.03) compared to their more efficient counterparts (7.1 vs. 6.6 kg DM/d). Average CH emissions were 202 and 222 g/d ( = 0.02) with the GEM system and 156 and 164 g/d ( = 0.40) with RC for the low- and high-RFI heifers, respectively. When adjusted for feed intake, CH yield (g/kg DMI) was similar for high- and low-RFI heifers (GEM: 27.7 and 28.5, = 0.25; RC: 26.5 and 26.5, = 0.99). However, CH yield differed between the 2 measurement techniques only for the high-RFI group ( = 0.01). Estimates of CO yield (g/kg DMI) also differed between the 2 techniques ( ≤ 0.03). Our study found that high- and low-efficiency cattle produce similar CH yield but different daily CH emissions. The 2 measurement techniques differ in estimating CH and CO emissions, partially because of differences in conditions (lower feed intakes of cattle while in chambers, fewer days measured in chambers) during measurement. We conclude that when intake of animals is known, the GEM system offers a robust and accurate means of estimating CH emissions from animals under field conditions.

Bacterial expansin-like proteins have synergistically increased cellulose hydrolysis by cellulolytic enzymes during the initial stages of biofuel production, but they have not been tested on livestock feeds. The objectives of this study were to: isolate and express an expansin-like protein (BsEXLX1), to verify its disruptive activity (expansion) on cotton fibers by immunodetection (Experiment 1), and to determine the effect of dose, pH and temperature for BsEXLX1 and cellulase to synergistically hydrolyze filter paper (FP) and carboxymethyl cellulose (CMC) under laboratory (Experiment 2) and simulated ruminal (Experiment 3) conditions. In addition, we determined the ability of BsEXLX1 to synergistically increase hydrolysis of corn and bermudagrass silages by an exogenous fibrolytic enzyme (EFE) (Experiment 4) and how different doses of BsEXLX1 and EFE affect the gas production (GP), in vitro digestibility and fermentation of a diet for dairy cows (Experiment 5). In Experiment 1, immunofluorescence-based examination of cotton microfiber treated without or with recombinant expansin-like protein expressed from Bacillus subtilis (BsEXLX1) increased the surface area by > 100% compared to the untreated control. In Experiment 2, adding BsEXLX1 (100 μg/g FP) to cellulase (0.0148 FPU) increased release of reducing sugars compared to cellulase alone by more than 40% (P < 0.01) at optimal pH (4.0) and temperature (50°C) after 24 h. In Experiment 3 and 4, adding BsEXLX1 to cellulase or EFE, synergistically increased release of reducing sugars from FP, corn and bermudagrass silages under simulated ruminal conditions (pH 6.0, 39°C). In Experiment 5, increasing the concentration of BsEXLX1 linearly increased (P < 0.01) GP from fermentation of a diet for dairy cows by up to 17.8%. Synergistic effects between BsEXLX1 and EFE increased in vitro NDF digestibility of the diet by 23.3% compared to the control. In vitro digestibility of hemicellulose and butyrate concentration were linearly increased by BsEXLX1 compared to the control. This study demonstrated that BsEXLX1 can improve the efficacy of cellulase and EFE at hydrolyzing pure substrates and dairy cow feeds, respectively.

Aflatoxin is a potent carcinogen often found in animal feedstuffs. Although it reportedly impairs development of the preimplantation pig embryo, it is not known whether it adversely affects development of the preimplantation bovine embryo. We conducted 3 experiments to investigate this possibility and determine whether deleterious effects of aflatoxin were caused by increased production of reactive oxygen species (ROS). Experiments were conducted with embryos produced in vitro and cultured after fertilization with various concentrations of aflatoxin. For experiment 1, embryos were treated with 0 (control), 40, 400, or 4,000 µg/L of aflatoxin B1 (AFB1). Treatment at all concentrations of AFB1 tended to reduce cleavage rate, with the 2 highest concentrations having significant effects. As compared with the control, 40 µg/L AFB1 reduced the percentage of oocytes becoming blastocysts and the percentage of cleaved embryos becoming blastocysts (19.7 vs. 8.1% and 30.3 vs. 14.3%, respectively). Complete inhibition of blastocyst formation occurred at concentrations of 400 and 4,000 µg/L of AFB1. Experiments 2 and 3 involved a 2 × 2 factorial design with effects of AFB1 (0 and 40 µg/L), the antioxidant Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, a water-soluble analog of vitamin E; 0 and 5 µM), and their interaction on production of ROS in putative zygotes (experiment 2) and development to the blastocyst stage (experiment 3). Production of ROS was increased by AFB1, and this effect was reversed by Trolox. However, Trolox did not prevent the reduction in development to the blastocyst stage caused by AFB1. Thus, the anti-developmental effects of AFB1 are not caused solely by increased ROS production. Rather, other underlying mechanisms exist for the adverse effects of aflatoxin on embryonic development. Overall, results indicate the potential for feeding aflatoxin-contaminated feed to cause embryonic loss in cattle.

The study applied 1H NMR-based plasma metabolomics to identify candidate biomarkers of aflatoxin B1 (AFB1) ingestion in dairy cows fed no sequestering agents and evaluate the effect of supplementing clay and/or a Saccharomyces cerevisiae fermentation product (SCFP) on such biomarkers. Eight lactating cows were randomly assigned to 1 of 4 treatments in a balanced 4 × 4 Latin square design with 2 squares. Treatments were: control, toxin (T; 1725 µg AFB1/head/day), T with clay (CL; 200 g/head/day), and CL with SCFP (CL + SCFP; 35 g of SCFP/head/day). Cows in T, CL, and CL + SCFP were dosed with AFB1 from d 26 to 30. The sequestering agents were top-dressed from d 1 to 33. On d 30 of each period, 15 mL of blood was taken from the coccygeal vessels and plasma samples were prepared by centrifugation. Compared to the control, T decreased plasma concentrations of alanine, acetic acid, leucine, arginine and valine. In contrast, T increased plasma ethanol concentration 3.56-fold compared to control. Treatment with CL tended to reduce sarcosine concentration, whereas treatment with CL + SCFP increased concentrations of mannose and 12 amino acids. Based on size of the area under the curve (AUC) of receiver operating characteristic and fold change (FC) analyses, ethanol was the most significantly altered metabolite in T (AUC = 0.88; FC = 3.56); hence, it was chosen as the candidate biomarker of aflatoxin ingestion in dairy cows fed no sequestering agent.

A 64-d receiving study was conducted to evaluate the effects of 3 feed additives on rate of bovine respiratory disease (BRD), fecal E. coli O157:H7 shedding, and performance of newly received steer calves. One hundred and forty-two crossbred steer calves (initial BW 295 ± 23 kg) from multiple sources (n=22) were utilized in this study. On d 0, steers were blocked by initial BW and assigned to 1 of 16 pens. Treatments consisted of basal diet with 114 g/hd/d of (1) placebo (Control), (2) direct-fed microbial blend, (3) direct-fed microbial/yeast extract blend, or (4) prebiotic/probiotic blend. No differences (P>0.05) in BRD incidence were observed in this study. A tendency (P=0.08) for a day by treatment interaction was observed for fecal E. coli O157:H7 prevalence with cattle consuming treatment 4 having less fecal E. coli O157:H7 on d 28 and d 56 relative to cattle on the control treatment. Dry matter intake, BW, ADG, and G:F were similar (P>0.10) among treatments. However, variability was reduced for steers on treatments 3 and 4 relative to steers on treatments 1 and 2 for d 64 DMI (SD = 2.8, 2.3, 1.2, and 1.8 kg for treatments 1, 2, 3, and 4 respectively) and BW (SD = 41.6, 38.7, 25.9, and 25.0 for treatments 1, 2, 3, and 4 respectively) and for ADG on d 28 (SD = 0.15, 0.15, 0.08, and 0.08 for treatments 1, 2, 3, and 4 respectively) and d 64 (SD = 0.30, 0.30, 0.08, and 0.09 for treatments 1, 2, 3, and 4 respectively). Results indicate that the cattle in this trial were not high risk for BRD. Although the feed additives used in this trial did not impact steer performance, 2 of the 3 additives reduced variability in steer performance, and 1 additive reduced fecal E. coli O157:H7 shedding.

The objective of this study was to evaluate the effects of a novel Lactobacillus plantarum strain with high-antioxidant activity on antioxidant status, chemical composition and fermentation profiles of alfalfa silage ensiled at two dry matter (DM) contents. Fresh chopped alfalfa was wilted to two targeted dry matter (DM) contents of approximately 300 (low DM) and 400 (high DM) g/kg fresh weight and treated without or with a strain of L. plantarum 24−7 with high antioxidant activity. Forages were ensiled in vacuum-sealed polyethylene bags containing ∼600 g wilted forage for 60 d, with 5 replicates for each treatment. Alfalfa silage ensiled at low DM had higher concentrations of acetic acid and non-protein nitrogen (NPN) and lower concentrations of lactic acid, propionic acid and water soluble carbohydrates (WSC) than silages with high DM. Inoculation of L. plantarum 24−7 led to lower silage pH and lower concentrations of NPN, neutral detergent fiber (aNDF), acid detergent fiber (ADF), and higher WSC compared to the control silage at two DM contents. Inoculation increased aerobic stability of the forage ensiled at low DM but decreased that ensiled at high DM. In control silages, almost half of the α-tocopherol and β-carotene was degraded during ensiling, whereas inoculation with L. plantarum 24−7 increased α-tocopherol and β-carotene concentrations as well as the proportion of polyunsaturated fatty acids (PUFA) in ensiled alfalfa at both DM contents. Greater concentrations of β-carotene were observed in L. plantarum 24−7 treated-silage at low DM compared to silage at high DM. The application of L. plantarum 24−7 strain also increased total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px) and catalase (CAT) activities of the alfalfa silage but decreased superoxide dismutase (SOD) and lipoxygenase (LOX) activities. These results indicate that L. plantarum 24−7 improved the fermentation quality and antioxidant status of ensiled alfalfa forage relative to the control silage.

The study objective was to evaluate the effects of sustained reduction of enteric methane (CH) emissions with dietary supplementation of the inhibitor 3-nitrooxypropanol (NOP) on growth rate and feed conversion efficiency of growing and finishing beef cattle. Eighty-four crossbred steers were used in a 238-d feeding study and fed a backgrounding diet for the first 105 d (backgrounding phase) and transition diets for 28 d followed by a finishing diet for 105 d (finishing phase) with 3 doses of NOP (0, 100, and 200 mg/kg DM). The experiment was a completely randomized design using 21 pens (4 cattle/pen) with 7 pens per treatment. When cattle were fed the backgrounding diet, pen DMI was reduced ( < 0.01) whereas G:F tended to improve ( = 0.06) with increasing dose of NOP supplementation. During the finishing phase, DMI ( = 0.06) and ADG ( = 0.07) tended to decrease with increasing dose of NOP supplementation. Although both levels of NOP were effective in reducing CH emissions from the backgrounding diet ( < 0.01), only NOP supplemented at the highest dose was effective in reducing total CH emissions from the finishing diet ( < 0.01). Methane yield (g/kg DMI) was reduced whereas hydrogen emissions were increased at the highest dose of NOP supplementation with both backgrounding and finishing diets ( < 0.01). Overall, these results demonstrate efficacy of NOP in reducing enteric CH emissions from cattle fed backgrounding and finishing diets, and these effects were negated once NOP supplementation was discontinued.

The forage lignocellulosic complex is one of the greatest limitations to utilization of the nutrients and energy in fiber. Consequently, several technologies have been developed to increase forage fiber utilization by dairy cows. Physical or mechanical processing techniques reduce forage particle size and gut fill and thereby increase intake. Such techniques increase the surface area for microbial colonization and may increase fiber utilization. Genetic technologies such as brown midrib mutants (BMR) with less lignin have been among the most repeatable and practical strategies to increase fiber utilization. Newer BMR corn hybrids are better yielding than the early hybrids and recent brachytic dwarf BMR sorghum hybrids avoid lodging problems of early hybrids. Several alkalis have been effective at increasing fiber digestibility. Among these, ammoniation has the added benefit of increasing the nitrogen concentration of the forage. However, few of these have been widely adopted due to the cost and the caustic nature of the chemicals. Urea treatment is more benign but requires sufficient urease and moisture for efficacy. Ammonia-fiber expansion technology uses high temperature, moisture, and pressure to degrade lignocellulose to a greater extent than ammoniation alone, but it occurs in reactors and is therefore not currently usable on farms. Biological technologies for increasing fiber utilization such as application of exogenous fibrolytic enzymes, live yeasts, and yeast culture have had equivocal effects on forage fiber digestion in individual studies, but recent meta-analyses indicate that their overall effects are positive. Nonhydrolytic expansin-like proteins act in synergy with fibrolytic enzymes to increase fiber digestion beyond that achieved by the enzyme alone due to their ability to expand cellulose microfibrils allowing greater enzyme penetration of the cell wall matrix. White-rot fungi are perhaps the biological agents with the greatest potential for lignocellulose deconstruction, but they require aerobic conditions and several strains degrade easily digestible carbohydrates. Less ruminant nutrition research has been conducted on brown rot fungi that deconstruct lignocellulose by generating highly destructive hydroxyl radicals via the Fenton reaction. More research is needed to increase the repeatability, efficacy, cost effectiveness, and on-farm applicability of technologies for increasing fiber utilization.

The objective of this study was to determine whether supplementing organic acids can prevent incidences of subacute ruminal acidosis (SARA) in beef heifers fed a diet consisting of 8% barley silage and 92% barley grain-based concentrate (DM basis). Ten ruminally cannulated Hereford crossbred heifers (484 ± 25 kg BW) were used in a replicated 5 × 5 Latin square design with 14-d periods including 10 d for dietary adaptation and 4 d for measurements. Dietary treatments included no supplementation (Control), low fumaric acid (61 g/d), high fumaric acid (125 g/d), low malic acid (59 g/d), and high malic acid (134 g/d). Organic acid supplementation had no effect on DMI ( = 0.77). Similarly, no effects were observed on mean ( = 0.74), minimum ( = 0.64), and maximum ( = 0.27) ruminal pH measured continuously for 48 h. Moreover, area under the curve for pH thresholds 6.2 ( = 0.97), 5.8 ( = 0.66), 5.5 ( = 0.55), and 5.2 ( = 0.93) was similar for all treatments. However, malic acid supplementation lowered the amount of time that ruminal pH was