Hugo A Ramirez- Ramirez

The use of portable X-ray fluorescence (PXRF) spectrometry to detect external markers on processed or unprocessed cattle and sheep fecal specimens to estimate apparent total tract digestibility (ATTD) was evaluated. Exp. 1: ruminally cannulated Angus-crossbred steers (n = 7; BW = 520 ± 30 kg) were individually fed ad libitum for 21 d in a completely randomized design (CRD). Markers (Cr₂O₃ and TiO₂) were placed inside the rumen twice daily (7.5 g of each marker). Fecal samples were collected twice daily from day 14 to 21. Exp. 2: crossbred wethers (n = 8; BW = 68 ± 3 kg) were individually fed ad libitum for 21 d in a CRD. During this period, 2 g of Cr₂O₃ and TiO₂ were top-dressed onto the feed twice daily. Sheep were housed in metabolism crates for 5 d for total fecal collection. Concentration of markers was determined on diets, refusals, and fecal specimens (fresh, dry-only, and dried/ground) using atomic absorption to detect Cr and spectrophotometry for Ti. Concentration of both markers was also determined via the PXRF spectrometer. Delta between ATTD estimated by wet chemistry and PXRF was not different from zero (P ≥ 0.14) when using cattle fresh fecal specimens for both markers, whereas ATTD estimated by PXRF with dry-only and dried/ground fecal specimens were 3.6 and 1.1 percent units lower (P ≤ 0.04), respectively, than ATTD estimated by wet chemistry for Cr and Ti, respectively. Regardless of the fecal sample preparation method on cattle specimens, Ti concentration was similar (P = 0.39) among methodologies, while Cr was underestimated (P < 0.01) by 13% when PXRF was used in dry-only or dried/ground samples. The ATTD of sheep was underestimated (P < 0.01) by 2.4 percent units compared with control when Cr was measured by PXRF in dry-only samples. The Cr concentration in dry-only fecal specimens of sheep tended (P = 0.09) to be lower compared with wet chemistry analysis. Fresh and dry/ground sheep fecal samples assessed for Cr, and dry-only assessed for Ti were not (P ≥ 0.49) affected by detection method. The Cr fecal recovery tended (P = 0.10) to be the lowest for dry-only, the greatest for wet chemistry, intermediate for fresh and dry/ground sheep-fecal specimens; while not affected (P = 0.40) for Ti. The PXRF is an accurate technology to detect Cr and Ti in fresh cattle fecal samples to estimate ATTD. For fresh and dry/ground, the technology was effective for determining the concentration of Cr, or dry-only fecal specimens when detecting Ti in sheep specimens.

The objectives of this study were to evaluate the effects of replacing 40 mg/kg of Zn from Zn sulfate (control; CON) with Zn AA complex (AvZn) on metabolism and immunological responses following an intravenous lipopolysaccharide (LPS) challenge in lactating cows. Cows were randomly assigned to 1 of 4 treatments: (1) pair-fed (PF) control (PF-CON; 5 mL of saline; n = 5), (2) PF AvZn (PF-AvZn; 5 mL of saline; n = 5), (3) LPS euglycemic clamp control (LPS-CON; 0.375 μg of LPS/kg of BW; n = 5), and (4) LPS euglycemic clamp AvZn (LPS-AvZn; 0.375 μg of LPS/kg of BW; n = 5). Cows were enrolled in 3 experimental periods (P). During period 1 (3 d), cows received their respective dietary treatments and baseline data were obtained. During period 2 (P2; 2 d), a 12-h LPS euglycemic clamp was conducted or cows were PF to their respective dietary counterparts. During period 3 (P3; 3 d), cows received their dietary treatment and consumed feed ad libitum. Mild hyperthermia (1°C) was observed in LPS cows at 3 h postbolus. Throughout P2, the rectal temperature of LPS-AvZn cows was decreased (0.3°C) relative to LPS-CON cows. Administrating LPS decreased dry matter intake (47%) during P2, and by experimental design the pattern was similar in PF cohorts. During P3, dry matter intake from LPS cows remained decreased (15%) relative to PF cows. Milk yield from LPS cows decreased (54%) during P2 relative to PF cows, but it was similar during P3. During P2, somatic cell count increased 3-fold in LPS cows relative to PF controls. Dietary AvZn tended to decrease somatic cell count (70%) during P3 relative to LPS-CON cows. Insulin increased 7-fold in LPS cows at 12 h postbolus and remained increased (4-fold) for the duration of P2. Circulating glucagon from LPS cows increased (65%) during P2, and supplementing AvZn blunted the increase (30% relative to LPS-CON). During P2, circulating cortisol increased 7-fold post-LPS infusion relative to PF cows, and supplementing AvZn decreased cortisol (58%) from 6 to 48 h postbolus relative to LPS-CON cows. Administrating LPS increased circulating LPS-binding protein and serum amyloid A (3- and 9-fold, respectively) relative to PF cows. Compared with LPS-CON, LPS-AvZn cows had increased circulating serum amyloid A (38%) 24 h postbolus. The 12-h total glucose deficit was 36 and 1,606 g for the PF and LPS treatments, respectively, but was not influenced by Zn source. In summary, replacing a portion of the Zn sulfate with Zn AA complex appeared to reduce the inflammatory response but had no effect on the glucose deficit.

Twenty Holstein cows, 12 primiparous and 8 multiparous, with (mean ± SD) 91 ± 19 d in milk and 595 ± 81 kg were used in replicated 4 × 4 Latin squares to compare the effects of feeding conventional dried distillers grains with solubles (DDGS) and reduced-fat DDGS (RFDDGS) in combination with rumen-inert fat (RIF) on milk production and rumen fermentation; one square contained rumen cannulated animals for rumen measurements. In each 21-d period, cows were randomly assigned to 1 of 4 dietary treatments (values on a dry matter basis): (1) control (CON) that contained 0% DDGS; (2) DG contained 30% DDGS; (3) RFDG contained 30% RFDDGS in substitution of DDGS; and (4) RFDG+RIF was similar to RFDG with the addition of 1.9% RIF. Unlike most practical diets in the dairy field, our diets had 18.0% crude protein. Dry matter intake was similar across treatments with any form of DDGS averaging 26.0 ± 0.6 kg/d, whereas the CON diet resulted in less dry matter intake, 21.6 ± 0.6 kg/d. Milk yield tended to be 1.7 kg/d greater for diets with either type of DDGS. Concentration of milk protein was greatest for the DG and RFDG diets, intermediate for the RFDG+RIF diet, and least for the CON diet, namely 3.22, 3.21, 3.12, and 3.07 ± 0.05%. Reduced milk fat percentage and yield were observed when cows consumed the DG diet, 3.27 ± 0.10% and 1.11 ± 0.04 kg/d, respectively, whereas these responses were similar among CON, RFDG, and RFDG+RIF, which averaged 3.68 ± 0.10% and 1.22 ± 0.04 kg/d. The presence of trans-10,cis-12 conjugated linoleic acid was only detected in milk from cows consuming the DG diet; similarly, concentration and yield of trans-10 18:1 were greater for cows consuming this diet. Rumen ammonia was similar across treatments averaging 27.0 ± 2.1mg/dL. The CON and RFDG+RIF diets had similar mean pH, 6.1 ± 0.11, whereas DG and RFDG resulted in lower pH averaging 5.79 ± 0.11. No effect on total concentration of volatile fatty acids was observed; the overall mean was 121 ± 4.11 mM; molar proportion of acetate was affected by treatment resulting in 67.3, 63.2, 61.4, and 60.9 ± 0.93 mol/100 mol for CON, RFDG+RIF, RFDG, and DG, respectively. Results from DNA sequencing showed that rumen bacterial community structure was relatively stable with minor changes at the family and genus levels; these changes may be associated with low starch diets, and hence reduced amylolytic bacteria populations. Feeding high proportions of RFDDGS resulted in greater dry matter intake with low risk for milk fat depression while supporting ruminal fermentation.

Fifteen primiparous crossbred dairy cows that were 114±14d in milk and weighed 533±56kg were used in a replicated 5×5 Latin square to test the efficacy of a calcium montmorillonite clay, NovaSil Plus (NSP; BASF Corp., Ludwigshaven, Germany), for the reduction of aflatoxin (AF) metabolite (AFM1) in milk and the effect of NSP on milk composition. Cows were housed in a freestall barn, fed once a day and milked twice a day. The experiment consisted of five 14-d periods: d 1 through 7 were considered for data collection, and d 8 through 14 were considered a wash-out phase. In each period, cows were randomly assigned to 1 of 5 dietary treatments: (1) control (CON), consisting of a basal total mixed ration (TMR); (2) high-dose NSP diet (NSP-1%), consisting of TMR plus 230 g of NSP; (3) aflatoxin diet (AFD), consisting of the TMR plus AF challenge; (4) low-dose NSP with AF (NSP-0.5%+AFD), composed of TMR plus 115 g of NSP and AF challenge; and (5) high-dose NSP with AF (NSP-1%+AFD), consisting of TMR plus 230 g of NSP and AF challenge. The AF challenge consisted of top dressing a daily dose of 100 µg/kg estimated dry matter intake (DMI); similarly, NSP was fed at 1.0 or 0.5% of estimated DMI. Milk yield and DMI were similar across treatments averaging 21.1±1.33 kg/d and 19.7±0.56 kg/d, respectively. Concentration of milk fat, protein, and lactose were similar across treatments with averages of 4.91±0.20%, 3.85±0.10%, and 4.70±0.06%, respectively. Concentration of vitamin A averaged 0.28±0.03 µg/mL and riboflavin concentration averaged 1.57±0.13 µg/mL across treatments. The concentration of minerals in milk were similar for all treatments. Cows fed CON and NSP-1% yielded the lowest concentration of AFM1 in milk with 0.03 and 0.01±0.06 µg/L. Addition of NSP reduced milk AFM1 from 1.10±0.06 µg/L with the AF diet to 0.58 and 0.32±0.06 µg/L with the NSP-0.5%+AF and NSP-1%+AF diets, respectively. Excretion of AFM1 was reduced by NSP; mean values were 24.38, 11.86, 7.38, 0.64, and 0.23, ± 1.71 µg/d, for AFD, NSP-0.5%+AFD, NSP-1%+AFD, NSP-1%, and CON, respectively. More specifically, 1.07±0.08% of the daily AF intake was transferred to the milk of cows consuming the AFD, whereas the AF transfer rates in milk from cows that consumed the NSP-0.5%+AFD and NSP-1%+AFD were 0.52 and 0.32±0.08%. Results from this research demonstrate that feeding NSP to lactating cows is an effective method to reduce the transfer and excretion of AFM1 in milk with no negative effects on dry matter intake, milk production, and composition.

Four ruminally cannulated Holstein cows averaging (± SD) 116 ± 18 d in milk and 686 ± 52 kg of body weight were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments to test the effects of forage particle size and concentration of corn oil on milk fat depression. Cows were housed in individual stalls, milked daily at 0700 and 1800 h, and individually fed daily at 0900 h for ad libitum consumption allowing approximately 10% orts. Four 28-d periods, in which each cow was offered 1 of 4 total mixed rations, included reduced-fat dried distillers grains with solubles at 30% of dietary dry matter and differed in forage particle size by inclusion of chopped grass hay (LONGP) or grass hay pellets (SHORTP) and 0 or 2% corn oil (CO). Dietary treatments were 0% corn oil + short particle size (CO0+SHORTP), 0% corn oil + long particle size (CO0+LONGP), 2% corn oil + short particle size (CO2 + SHORTP), and 2% corn oil + long particle size (CO2 + LONGP). Dry matter intake and milk yield were not affected by treatment averaging 26.5 ± 1.19 kg/d and 32.8 ± 3.34 kg/d, respectively. A decrease was found in 3.5% fat-corrected milk with the inclusion of oil resulting in 34.6 and 26.6 ± 2.6 kg/d for 0 and 2% oil diets, respectively. An oil × size interaction was found for milk fat concentration resulting in 2.27, 3.02, 3.62, and 3.62 ± 0.23% for CO2+SHORTP, CO2 + LONGP, CO0 + SHORTP, and CO0 + LONGP, respectively. Fat yield was reduced from 1.22 to 0.81 ± 0.09 kg/d with 2% oil diets. Cows consuming diets with long particle size spent 29 more minutes eating compared with the cows consuming short particle size (198 and 169 ± 15 min/d). Rumination time decreased from 504 to 400 ± 35 min/d for cows consuming short particle size compared with long particle size. Total chewing was reduced from 702 to 570 ± 4 min/d when cows consumed short particle size. Feeding long particle size decreased rate of passage of dry matter from 3.38 to 2.89 ± 0.42%/h; concomitantly mean retention time increased from 31.7 to 38.4 ± 5.36 h for diets containing long particle size. The results of this experiment show that effects of oil on milk fat depression were less severe when cows consumed long particle size, suggesting that dietary manipulations that modify rumen kinetics also affect milk fat production in dairy cows consuming reduced-fat dried distillers grains with solubles supplemented with corn oil.

Two experiments were conducted to evaluate the additive effects of starch and fat as risk factors associated with milk fat depression in dairy diets containing corn dried distillers grains with solubles. In experiment 1, 4 multiparous ruminally cannulated Holstein cows, averaging 114 14 d in milk and 662 52 kg of body weight, were randomly assigned to 4 treatments in a 4 x 4 Latin square to determine the effect of these risk factors on rumen fermentation and milk fatty acid profile. In each 21-d period, cows were assigned to 1 of 4 dietary treatments: a control diet (CON; ether extract 5.2%, starch 19%); CON with added oil (OL; ether extract 6.4%, starch 18%); CON with added starch (STR; ether extract 5.5%, starch 22%); and CON with added oil and starch (COMBO; ether extract 6.5%, starch 23%). After completion of experiment 1, milk production response was evaluated in a second experiment with a similar approach to diet formulation. Twenty Holstein cows, 12 primiparous and 8 multiparous, averaging 117 17 d in milk and 641 82 kg, were used in replicated 4 x 4 Latin squares with 21-d periods. Results from experiment 1 showed that ruminal pH was not affected by treatment averaging 5.87 0.08. Molar proportion of propionate in rumen fluid was greatest on the COMBO diet, followed by OL and STR, and lowest for CON. The concentration of trans-10,cis-12 conjugated linoleic acid in milk fat increased with the COMBO diet. Adding oil, starch, or a combination of both resulted in lower concentration and yield of fatty acids <16 carbons. Compared with the control, OL and STR resulted in 13% lower concentration, whereas the COMBO diet resulted in a 27% reduction; similarly yield was reduced by 24% with the OL and STR treatments and 54% with the COMBO diet. In experiment 2, milk yield, milk protein percentage, and milk protein yield were similar across treatments, averaging 26.6 +/- 1.01 kg/d, 3.2 +/- 0.05%, and 0.84 +/- 0.03 kg/d, respectively. Fat-corrected milk was greatest for CON, 26.5 1.12 kg/d; no differences were detected among the remaining treatments, which averaged 23.5 +/- 1.12 kg/d. Milk fat percentage was greatest when cows consumed CON, 3.3 +/- 0.15%; OL and STR averaged 3.0 +/- 0.15% and COMBO resulted in the lowest milk fat percentage, 2.73 +/- 0.15%. Milk fat yield was 0.25 +/- 0.05 kg/d greater for the CON diet compared with the other 3 treatments, which were similar. These results suggest that fat and starch are additive risk factors that will likely induce milk fat depression in diets containing high inclusion of dried distillers grains with solubles.

Corn grain and corn silage are major feed components in lactating dairy cow rations. Bacillus thuringiensis (B.t.) is a naturally occurring soil bacterium that produces a protein that is toxic to lepidopteran insects that may damage plant tissues and reduce corn quality and yields. During each of the four 28-d periods, cows were offered 1 of 4 rations in which the corn grain and silage originated from different corn hybrids: a non-transgenic corn control (from hybrid DKC63-78; Monsanto Co., St. Louis, MO), a B.t. test substance corn (MON 89034 in hybrid DKC63-78; Monsanto Co.), and 2 commercial nontransgenic reference (Ref) hybrids: DKC61-42 (Ref 1) and DKC62-30 (Ref 2; Monsanto Co.). Sixteen multiparous Holstein cows averaging 110 21 d in milk and weighing 684 +/- 62.3 kg were blocked by days in milk and milk yield and randomly assigned to one of four 4 x 4 Latin squares. Diets were formulated to contain 36.4% corn silage and 16.3% corn grain. Dry matter intake was greater for cows consuming B.t. corn (26.6 +/- 0.59 kg/d) compared with the control, Ref 1, and Ref 2 corn diets (25.4, 25.0, and 25.6 0.59 kg/d, respectively). Milk yield, fat yield, and percentage of fat (36.8 +/- 0.98 kg/d, 1.22 +/- 0.05 kg/d, and 3.3 +/- 0.10%), milk protein yield and percentage of protein (1.11 +/- 0.03 kg/d and 3.01 +/- 0.05%), milk urea nitrogen concentration (14.01 +/- 0.49 mg/dL), and 3.5% fat-corrected milk yield (35.7 +/- 1.07 kg/d) were not different across treatments. The results from this study show that lactating dairy cows that consume B.t. corn (MON 89034) do not differ from lactating dairy cows that consume nontransgenic corn in milk yield, 3.5% fat-corrected milk per unit of dry matter intake, or milk components.

Sixteen multiparous lactating Holstein cows were used in 2 experiments to evaluate the effects of reduced-fat dried distillers grains with solubles (RFDG) on milk production, rumen fermentation, intestinal microbial N flow, and total-tract nutrient digestibility. In experiment 1, RFDG was fed at 0, 10, 20, or 30% of diet dry matter (DM) to 12 noncannulated Holstein cows (mean ± standard deviation: 89 ± 11 d in milk and 674 ± 68.2 kg of body weight) to determine effects on milk production. In experiment 2, the same diets were fed to 4 ruminally and duodenally cannulated Holstein cows (mean ± standard deviation: 112 ± 41 d in milk; 590 ± 61.14 kg of body weight) to evaluate the effects on rumen fermentation, intestinal flow of microbial N, and total-tract nutrient digestibility. In both experiments, cows were randomly assigned to 4 × 4 Latin squares over 21-d periods. Treatments (DM basis) were (1) control (0% RFDG), (2) 10% RFDG, (3) 20% RFDG, and (4) 30% RFDG. Feed intake and milk yield were recorded daily. In both experiments, milk samples were collected on d 19 to 21 of each period for analysis of milk components. In experiment 2, ruminal pH was measured; samples of rumen fluid, duodenal digesta, and feces were collected on d 18 to 21. Microbial N was estimated by using purines and DNA as microbial markers. Milk yield was not affected by treatment and averaged 34.0 ± 1.29 kg/d and 31.4 ± 2.81 kg/d in experiments 1 and 2, respectively. Percentage of milk protein tended to increase in experiment 1; estimates were 3.08, 3.18, 3.15, and 3.19 ± 0.06% when RFDG increased from 0 to 30% in the diets. However, milk protein concentration was not affected in experiment 2 and averaged 3.02 ± 0.07%. Percentage of milk fat was not affected and averaged 3.66 ± 0.05% and 3.25 ± 0.14% in experiments 1 and 2, respectively. Total ruminal volatile fatty acids and ammonia concentrations were not affected by treatment and averaged 135.18 ± 6.45 mM and 18.66 ± 2.32 mg/dL, respectively. Intestinal microbial N flow was not affected by treatment; however, purines yielded higher estimates of flow compared with DNA markers. When averaged across treatments, intestinal flow of microbial N was 303 and 218 ± 18 g of N/d, using purines and DNA as the markers. Dry matter, organic matter, neutral detergent fiber, and nonfiber carbohydrate digestibility tended to increase with increasing inclusion of RFDG. Results from these experiments indicate that dairy rations can be formulated to include up to 30% RFDG while maintaining lactation performance, volatile fatty acids concentration, and intestinal supply of microbial N.

The objectives of this study were to evaluate the effect of dried distillers grains with solubles (DDGS) on ruminal biohydrogenation and duodenal flow of fatty acids, and to evaluate effects on the ruminal and duodenal microbial community using Roche 454 pyro-sequencing. Three crossbred steers (average BW 780 ± 137 kg) fitted with ruminal and duodenal cannulae were used in a 3-diet, 6-period crossover design. Animals were housed in individual free stalls and fed twice daily at 0700 and 1900 h. Diets (DM basis) were 1) CONTROL, 19.5% corn bran, 20% sorghum silage, 60% brome hay, 0.5% trace minerals, and 0.25% urea, but no DDGS; 2) LOW DDGS, inclusion of 9.75% DDGS replacing equal percentage of corn bran; 3) HIGH DDGS, inclusion of 19.5% DDGS completely replacing corn bran. Feed ingredients and duodenal digesta samples were analyzed for fatty acid composition. The DNA was extracted from isolated mixed ruminal bacterial samples and from intestinal digesta samples. The V1-V3 region of the 16S rRNA gene was sequenced, and bacterial phylogenetic analysis was conducted. Data were analyzed using the MIXED procedure of SAS. Biohydrogenation of C18:1 increased (P < 0.01) with DDGS inclusion; means were 68.3, 75.6, and 79.3 ± 4.3% for CONTROL, LOW DDGS, and HIGH DDGS, respectively. In the same order, means of biohydrogenation of C18:2 (P < 0.05) were 84.1, 91.5, and 93.3 ± 3.4%. Duodenal flow of total fatty acids increased (P < 0.01) with DDGS inclusion; means were 134, 168, and 223 ± 33 g/d for CONTROL, LOW DDGS, and HIGH DDGS, respectively. In the same order, means of C18:0 flow (P < 0.01) were 51, 86, and 121 ± 18 g/d. DDGS did not affect the predominant bacterial phyla in the gut, which were Bacteroidetes (P = 0.62) and Firmicutes (P = 0.71). However, the phylum Fibrobacteres decreased (P < 0.01) when DDGS was fed with means of 5.5, 6.0 and 3.7 ± 0.6% for CONTROL, LOW DDGS, and HIGH DDGS, respectively. Fibrobacteres were lower (P < 0.01) in isolated ruminal bacterial samples compared to duodenal digesta samples with means of 0.1 and 10.1 ± 0.6%, respectively. Overall, the inclusion of DDGS in diets increased ruminal biohydrogenation of C18:1 and C18:2, which increased duodenal flow of C18:0. In addition, the bacterial community of the rumen clustered separately from that of the duodenum suggesting different bacterial diversity between isolated ruminal bacteria and duodenal digesta.

Paz, H. A., Castillo-Lopez, E., Ramirez-Ramirez, H. A., Christensen, D. A., Klopfenstein, T. J. and Kononoff, P. J. 2013. Invited Review: Ethanol co-products for dairy cows: there goes our starch … now what? Can. J. Anim. Sci. 93: 407–425. The rise of the grain-ethanol industry has resulted in a dramatic increase in the availability of feed co-products namely, distillers’ grains with solubles (DG). The cost of feeds that have traditionally been used for energy continues to increase and there is a need to understand the potential impact of replacing these feeds with DG on milk yield and composition. Using the Dairy National Research Council (2001) model and data from a total of 25 published studies (81 observations), we used a meta-analytical procedure to evaluate the impact of feeding corn milling co-products on lactating dairy cows. Based on our model, the inclusion of DG did not affect dry matter intake (P=0.35) but a trend for an increase in milk yield was observed (P=0. 10). Additionally, the percentages of milk fat and protein in the milk were not affected (P=0.24 and 0.25, respectively). The modeled daily rumen outflow of Lys in grams was significantly (P−1and was not affected by DG (P=0.79) but the error associated with metabolizable protein allowable milk tended (P=0.10) to be higher for cows consuming DG by 1.4±1.2 kg d⁻¹.