James S. Drouillard

Daily Injection of Tumor Necrosis Factor-α Increases Hepatic Triglycerides and Alters Transcript Abundance of Metabolic Genes in Lactating Dairy Cattle

To determine whether inflammation can induce bovine fatty liver, we administered recombinant bovine tumor necrosis factor-alpha (rbTNF) to late-lactation Holstein cows. Cows (n = 5/treatment) were blocked by feed intake and parity and randomly assigned within block to control (CON; saline), rbTNF at 2 microg/(kg.d), or pair-fed control (saline, intake matched) treatments. Treatments were administered once daily by subcutaneous injection for 7 d. Plasma samples were collected daily for analysis of glucose and FFA and a liver biopsy was collected on d 7 for triglyceride (TG) and quantitative RT-PCR analyses. Data were analyzed using treatment contrasts to assess effects of tumor necrosis factor-alpha (TNFalpha) and decreased feed intake. By d 7, feed intake of both rbTNF and pair-fed cows was approximately 15% less than CON (P < 0.01). Administration of rbTNF resulted in greater hepatic TNFalpha mRNA and protein abundance and 103% higher liver TG content (P < 0.05) without affecting the plasma FFA concentration. Hepatic carnitine palmitoyltransferase 1 transcript abundance tended to be lower (P = 0.09) and transcript abundance of fatty acid translocase and 1-acyl-glycerol-3-phosphate acyltransferase was higher (both P < 0.05) after rbTNF treatment, consistent with increased FFA uptake and storage as TG. Transcript abundance of glucose-6-phosphatase (P < 0.05) and phosphoenolpyruvate carboxykinase 1 (P = 0.09), genes important for gluconeogenesis, was lower for rbTNF-treated cows. These findings indicate that TNFalpha promotes liver TG accumulation and suggest that inflammatory pathways may also be responsible for decreased glucose production in cows with fatty liver.

Effects of Dried Distillers' Grain on Fecal Prevalence and Growth of Escherichia coli O157 in Batch Culture Fermentations from Cattle†

ABSTRACT Distillers’ grains (DG), a by-product of ethanol production, are fed to cattle. Associations between Escherichia coli O157 prevalence and feeding of DG were investigated in feedlot cattle (n = 379) given one of three diets: steam-flaked corn (SFC) and 15% corn silage with 0 or 25% dried distillers’ grains (DDG) or SFC with 5% corn silage and 25% DDG. Ten fecal samples were collected from each pen weekly for 12 weeks to isolate E. coli O157. Cattle fed 25% DDG with 5 or 15% silage had a higher (P = 0.01) prevalence of E. coli O157 than cattle fed a diet without DDG. Batch culture ruminal or fecal microbial fermentations were conducted to evaluate the effect of DDG on E. coli O157 growth. The first study utilized microbial inocula from steers fed SFC or dry-rolled corn with 0 or 25% DDG and included their diet as the substrate. Ruminal microbial fermentations from steers fed DDG had higher E. coli O157 contents than ruminal microbial fermentations from steers fed no DDG (P < 0.05) when no substrate was included. Fecal fermentations showed no DDG effect on E. coli O157 growth. In the second study with DDG as a substrate, ruminal fermentations with 0.5 g DDG had higher (P < 0.01) E. coli O157 concentrations at 24 h than ruminal fermentations with 0, 1, or 2 g DDG. In fecal fermentations, 2 g DDG resulted in a higher concentration (P < 0.05) at 24 h than 0, 0.5, or 1 g DDG. The results indicate that there is a positive association between DDG and E. coli O157 in cattle, and the findings should have important ramifications for food safety.

Impact of beef cattle diets containing corn or sorghum distillers grains on beef color, fatty acid profiles, and sensory attributes

Strip loins from 236 carcasses from crossbred yearling steers were collected on each of 2 slaughter dates (slaughter 1 or 2) to determine the effects of feeding corn or sorghum distillers grains (DG) on beef color, fatty acid profiles, lipid oxidation, tenderness, and sensory attributes. Dietary treatments consisted of a steam-flaked corn (SFC) diet without (control) or with 15% (DM basis) corn dry or wet DG (CDDG and CWDG) or sorghum dry or wet DG (SDDG and SWDG) and alfalfa hay (R). Additional treatments included SDDG or SWDG with no alfalfa hay (NR). In slaughter 2, steaks from steers fed SFC had lesser L*, but greater a* (P < 0.05) values than those from steers fed DG. When comparing sorghum and corn DG steaks, the same color differences were detected. Steaks from steers fed sorghum DG had lower L*, but greater a* (P < 0.05) values than those from steers fed corn DG. Also, L* values in steaks from steers fed SWDG with R were greater (P < 0.05) than those from steers fed SWDG with NR. In slaughter 1, feeding DG increased (P < 0.05) steak n-6 fatty acid concentrations compared with SFC. In both slaughter groups, feeding dry DG increased (P < 0.05) steak linoleic acid concentrations compared with wet DG. In slaughter 2, feeding corn DG diets increased (P < 0.05) linoleic acid concentrations of steaks compared with sorghum DG diets. In addition, increased (P < 0.05) concentrations of alpha-linolenic acid in steaks resulted from feeding SDDG or SWDG with R compared with those sorghum treatments with NR. In each slaughter group, feeding DG increased (P < 0.05) the n-6:n-3 ratio of steaks compared with SFC, and feeding corn DG increased (P < 0.05) this ratio compared with sorghum DG. Furthermore, steaks from steers fed corn DG had greater (P < 0.05) concentrations of trans-vaccenic acid than those from steers fed sorghum DG. In slaughter 1, the CLA isomer 18:2, trans-10, cis-12 was greater (P < 0.05) in steaks from DG diets. On d 1 of retail display, steaks from steers fed SDDG with R in slaughter 2 had greater (P < 0.05) thiobarbituric acid reactive substances values than those from steers fed SDDG with NR. Feeding DG at 15% of the dietary DM did not affect sensory attributes or Warner-Bratzler shear force values of steaks. Feeding DG from either corn or sorghum as either a wet or dry by-product had no effect on beef sensory attributes.

Efficacy of Escherichia coli O157:H7 Siderophore Receptor/Porin Proteins–Based Vaccine in Feedlot Cattle Naturally Shedding E. coli O157

Escherichia coli O157 is a foodborne pathogen commonly isolated from beef cattle feces and can enter the food chain at harvest. A relatively new vaccine technology uses the iron requirement of pathogenic bacteria by targeting the siderophore receptor and porin proteins (SRP). We evaluated the efficacy of an anti-E. coli O157 SRP-based vaccine in feedlot cattle naturally shedding the organism. Sixty cattle were selected from an original population of 600; 50 of these were fecal positive for E. coli O157 on two occasions and the remaining 10 animals were fecal positive on one occasion. Cattle were stratified based on the results of screening samples and randomly allotted to one of three treatment groups: control, vaccinated with 2 mL per animal of E. coli SRP vaccine, or vaccinated with 3 mL per animal of E. coli SRP vaccine subcutaneously 21 days apart. Control cattle were injected with sterile saline emulsified with an adjuvant. Fecal samples and rectoanal mucosal swab samples were collected two or three times a week for 8 weeks to monitor shedding of E. coli O157. Prevalence of the pathogen was analyzed by repeated measures on animals over weeks. The SRP vaccine at the 3 mL dose reduced prevalence of E. coli O157 compared to the control (17.7% vs. 33.7%; p < 0.01). A similar trend was observed with the 2 mL dose (29.1%), but differences were not statistically significant compared to control (p = 0.40). Additionally, the 3 mL dose of SRP vaccine reduced the number of days cattle tested culture positive for E. coli O157 (p = 0.05) and the number of days cattle were identified as high-shedders (p = 0.02) compared to control.

Effect of feedlot management system on response to ractopamine-HCl in yearling steers.

Two experiments evaluated the effects of conventional and natural feedlot management systems (MS) on ractopamine-HCl (RAC) response in yearling steers. Feedlot performance, carcass characteristics, skeletal muscle gene expression, and circulating IGF-I concentrations were measured. The conventional system included a combined trenbolone acetate and estradiol implant, Revalor-S (IMP), as well as monensin-tylosin feed additives (IA). Treatments were arranged in a 2 x 2 factorial and included: 1) natural (NAT): no IMP-no IA, no RAC; 2) natural plus (NAT+): no IMP-no IA, RAC; 3) conventional (CON): IMP-IA, no RAC; and 4) conventional plus (CON+): IMP-IA, RAC. In Exp. 1, one hundred twenty crossbred steers (initial BW = 400 +/- 26 kg) were allotted randomly to treatment in a randomized complete block design (BW was blocking criteria); pen was the experimental unit. In Exp. 2, twenty-four individually fed crossbred steers (initial BW = 452 +/- 25 kg) were used in a randomized complete block design (BW was blocking criteria) and assigned to the same treatments as Exp. 1, with 6 steers/treatment. In Exp. 2, serum was harvested on d 0 and 31 and within the 28-d RAC feeding period, at d 0, 14, and 28. Longissimus biopsy samples were taken on d 0, 14, and 28 of the RAC feeding period for mRNA analysis of beta-adrenergic receptors and steady-state IGF-I mRNA. In Exp. 1, ADG, G:F, final BW, and HCW were greatest for CON+ (P < 0.01). During the final 37 d, RAC increased ADG (P = 0.05) and increased overall G:F (P = 0.02). Marbling score was reduced (P = 0.02), and yield grade was improved with RAC (P = 0.02), but RAC did not affect dressing percentage (P = 0.96) or HCW (P = 0.31). In Exp. 2, MS x RAC interactions were detected in ADG and G:F the last 28 d, overall ADG and overall G:F, final BW, and HCW (P < 0.01). Dressing percentage, yield grade, and marbling score were not altered by MS or RAC (P > 0.10). Circulating IGF-I concentration was increased on d 31 by the conventional MS, and concentration was greater throughout the study than NAT steers (P < 0.01). Circulating IGF-I concentrations were not changed by RAC (P = 0.49). Abundance of beta(1)-AR mRNA tended to increase (P = 0.09) with RAC, but RAC did not affect beta(2)-AR, beta(3)-AR, or IGF-I mRNA (P > 0.40). Management system did not affect beta(1)-AR, beta(2)-AR, beta(3)-AR, or IGF-I mRNA (P > 0.18), yet a trend (P = 0.06) for MS x RAC for beta(2)-AR mRNA was detected. These results indicate that response to RAC is affected by feedlot management practices.

Occurrence of the transferable copper resistance gene tcrB among fecal enterococci of U.S. feedlot cattle fed copper-supplemented diets.

Copper, an essential micronutrient, is supplemented in the diet at elevated levels to reduce morbidity and mortality and to promote growth in feedlot cattle. Gut bacteria exposed to copper can acquire resistance, which among enterococci is conferred by a transferable copper resistance gene (tcrB) borne on a plasmid. The present study was undertaken to investigate whether the feeding of copper at levels sufficient to promote growth increases the prevalence of the tcrB gene among the fecal enterococci of feedlot cattle. The study was performed with 261 crossbred yearling heifers housed in 24 pens, with pens assigned randomly to a 2�2 factorial arrangement of treatments consisting of dietary copper and a commercial linseed meal-based energy protein supplement. A total of 22 isolates, each identified as Enterococcus faecium, were positive for tcrB with an overall prevalence of 3.8% (22/576). The prevalence was higher among the cattle fed diets supplemented with copper (6.9%) compared to normal copper levels (0.7%). The tcrB-positive isolates always contained both erm(B) and tet(M) genes. Median copper MICs for tcrB-positive and tcrB-negative enterococci were 22 and 4 mM, respectively. The transferability of the tcrB gene was demonstrated via a filter-mating assay. Multilocus variable number tandem repeat analysis revealed a genetically diverse population of enterococci. The finding of a strong association between the copper resistance gene and other antibiotic (tetracycline and tylosin) resistance determinants is significant because enterococci remain potential pathogens and have the propensity to transfer resistance genes to other bacteria in the gut.

Evaluation of feeding dried distiller's grains with solubles and dry-rolled corn on the fecal prevalence of Escherichia coli O157:H7 and Salmonella spp. in cattle.

Escherichia coli O157:H7 and Salmonella are foodborne pathogens that reside in the gut of cattle and are shed in the feces. Previous work indicated a positive association between feeding cattle distillers grains (DG) and an increase in E. coli O157:H7 prevalence. Feeding processed grains also has been shown to affect fecal prevalence of E. coli O157:H7. The objective of this study was to evaluate the effect of feeding DG and dry-rolled corn (DRC), alone or in combination, on fecal prevalence of E. coli O157:H7 and Salmonella in finishing cattle. Cattle were allotted to pens (n = 28), and fed dietary treatments (n = 150 days) structured in a 2 x 2 factorial arrangement; the factors were 0% or 25% dried corn DG with solubles (DDGS) and 0% or 25% DRC in steam-flaked corn-based high-grain diets. Fecal samples were collected from each pen floor before initiating dietary treatments and at least once every 2 weeks after final diets began. Overall prevalence of E. coli O157:H7 in fecal samples was 5.1%. There were no significant effects of DDGS, DRC, or sampling time on E. coli O157:H7 prevalence (p > 0.20). Overall prevalence of Salmonella in pen floor fecal samples was 23.7%, and sampling week affected prevalence (p < 0.01), ranging from < 1% (week 1) to 77.5% (week 17). Salmonella prevalence was not affected by cattle diet, and no work had previously reported an association between either DG or DRC and Salmonella prevalence. Lack of an association between E. coli O157:H7 prevalence and feeding DG or DRC is contrary to previous observations. Further research is needed to understand inconsistencies between studies of E. coli O157:H7 prevalence and potential associations with DG and grain-processing methods.

Effects of feeding elevated concentrations of copper and zinc on the antimicrobial susceptibilities of fecal bacteria in feedlot cattle.

Cattle are fed elevated concentrations of copper and zinc for growth promotion. The potential mechanisms of growth promotional effects of these elements are attributed to their antimicrobial activities, similar to that of antibiotics, in that gut microbial flora are altered to reduce fermentation loss of nutrients and to suppress gut pathogens. Copper and zinc fed at elevated concentrations may select for bacteria that are resistant not only to heavy metals but also to antibiotics. Our objectives were to determine the effects of feeding elevated copper and zinc on the antimicrobial susceptibilities of fecal bacteria in feedlot cattle. Twenty heifers, fed corn-based high-grain diets, were randomly assigned to treatments in a 2 x 2 factorial arrangement with 1X or 10X National Research Council recommended copper and/or zinc. Feces, collected on days 0, 14, and 32, were cultured for commensal bacteria (Escherichia coli and Enterococcus) to determine their susceptibilities to copper, zinc, and antibiotics. Fecal DNA was extracted to detect tcrB gene and quantify erm(B) and tet(M) genes. In E. coli and Enterococcus sp., minimal differences in minimum inhibitory concentrations (MICs) of copper, zinc, and antibiotics were noticed. The mean copper MIC for E. coli increased (p < 0.05) between days 0 and 32 and days 14 and 32. The only treatment effect detected was increased zinc MIC of E. coli isolates (p < 0.01). The tcrB gene was not detected in feces or in enterococcal isolates. Proportions of erm(B) and tet(M) were unaffected by copper or zinc supplementation. However, the proportion of tet(M) increased (p < 0.05) between days 0 and 14. Feeding elevated copper and/or zinc to feedlot cattle had marginal effects on antimicrobial susceptibilities of fecal E. coli and enterococci.

Effects of base ingredient in cooked molasses blocks on intake and digestion of prairie hay by beef steers.

Twelve steers (332 kg) were used in three simultaneous 4 x 3 incomplete Latin squares to evaluate effects of beet molasses (BEET), cane molasses (CANE), or concentrated separator by-product (CSB) as base ingredients in cooked molasses blocks on intake and digestion of prairie hay and ruminal characteristics. All steers had ad libitum access to prairie hay (5.9% CP and 69.4% NDF; DM basis). The four experimental treatments included a control (no supplement) and three cooked molasses blocks, based on BEET, CANE, or CSB, fed daily at .125% of BW (.42 kg/d as-fed, .13 kg/d CP). Forage OM, NDF, and N intakes; digestible OM, NDF, and N intakes; and total tract OM and N digestibilities (percentage of intake) were greater (P < .05) for steers fed cooked molasses blocks than for control steers. Total tract OM digestibility was greater (P < or = .06) for steers fed BEET blocks (54.0%) than for those fed CSB (52.1%) or CANE blocks (52.2%). Digestion of NDF was greatest (P < .05) for steers fed BEET blocks (51.9%) and tended to be greater (P < .07) for steers fed CANE (49.3%) or CSB blocks (49.3%) than for control steers (46.9%). Ruminal ammonia concentrations were greater (P < .05) for steers fed cooked molasses blocks (.89 mM) than for control steers (.21 mM); this was primarily due to increases to 4.6 mM at 2 h postfeeding for steers fed blocks. Concentrations of total VFA in ruminal fluid were greater (P < .05) for steers fed BEET (92.7 mM) and CSB (88.1 mM) blocks than for control steers (80.3 mM), whereas concentrations for steers fed CANE blocks were intermediate (85.4 mM). Steers supplemented with cooked molasses blocks had greater molar percentages of butyrate than did control steers, particularly shortly after feeding. In summary, supplementation with cooked molasses blocks increased forage intake and digestion. The three base ingredients elicited similar responses, although steers fed BEET had slightly greater OM and NDF digestibilities than those fed CANE or CSB.

Wet distiller’s grains with solubles in beef finishing diets comprised of steam-flaked or dry-rolled corn

The purpose of this experiment was to determine optimal levels of distiller’s grains in finished diets with steam-flaked corn or dry rolled corn. Distiller’s grains have been used extensively in regions of the country in which dry-rolled and high-moisture grains are prevalent. Production of fuel ethanol is now expanding into the High Plains, where feedlots more commonly use steam flaking. The cost to produce flaked corn is higher than the cost to produce dry rolled corn, and with rising energy costs (especially natural gas), this spread is becoming more dramatic. Comparing the use of wet distiller’s grains fed in conjunction with these grains provides useful information concerning optimum use level.