Fecal Grab Samples Research Articles

Evaluation of two external markers for measurement of ileal and total tract digestibility of pigs fed human-type diets1

External markers for determination of nutrient digestibility have often been evaluated in conventional dry feeds but less often in conventional feeds such as human-type diets used in animal model studies. In the present study, 5 ileal-cannulated pigs were fed 5 types of soft bread-based diets supplemented with Cr(2)O(3) and AIA as digestibility markers for 1 wk in a Latin square design. Ileal contents were collected twice for 5 h and a fecal grab sample was obtained once per week. Ileal and total tract digestibility of OM and nonstarch polysaccharides (NSP) based on the 2 markers were compared by linear regression. Across dietary treatments and site of collection, high correlation existed between digestibility values obtained with Cr(2)O(3) and AIA, resulting in a R(2) > 0.77 (P < 0.001) and a linear relation close to unity. For ileal samples, the correlation was poor, particularly for NSP, which had R(2) = 0.09 (P = 0.14) whereas OM had an R(2) = 0.52 (P <0.001). On the other hand, fecal grab samples led to R(2) > 0.92 (P < 0.001) for both OM and NSP. However, AIA gave higher values than Cr(2)O(3), particularly in samples with lower digestibility. The discrepancy is presumably caused by analytical difficulties due to a high fecal ash contents or interference with other components in the human-type diets.

Effects of cecal oxytetracycline infusion, and dietary avidin and biotin supplementation on the biotin status of nongravid gilts1

The objective of this 49-d experiment was to test effects of cecal oxytetracycline (OTC) infusion, and dietary avidin and biotin supplementation on the biotin status of nongravid gilts. Twenty-eight crossbred gilts with an initial age of 160 d and BW of 120 kg were surgically fitted with a T-cannula in the terminal ileum, a cecal fistula, and an indwelling catheter in the anterior vena cava, and allotted to 7 dietary treatments. Treatments with the basal semipurified (SP) diet fed at 1.86 kg/d were: SP-1, negative control; SP-2, positive control with 270 μg of biotin/kg; SP-3, with spray-dried egg albumen (EA, 100 g/d) and OTC (2.56 g/d by cecal infusion); and SP-4, with EA, OTC, and 700 μg of biotin/kg. Treatments with the basal corn-soybean meal (CS) diet fed at 1.80 kg/d were: CS-1, negative control; CS-2, with EA and OTC; and CS-3, with EA, OTC, and 700 μg of biotin/kg. Response criteria were: fecal bacteria counts; plasma concentrations of biotin, glucose, and urea N (PUN); liver pyruvate carboxylase (PC) activity; kidney and epithelial tissue histology; ileal and fecal biotin concentrations; ileal and total tract N and energy utilization; daily gilt observation; and BW gain. Blood samples were collected every 7 d with serial samples collected on d 49. Total urine collections and fecal grab samples were made twice daily from d 44 to 49. Gilts were killed on d 50 and liver, kidney, and skin samples were collected. No gilts had symptoms of biotin deficiency. There were no treatment differences in BW gain, plasma glucose concentrations, liver PC activity, kidney and epithelial tissue histology, or fecal bacteria counts. Ileal and total tract N and energy digestibilities (%) did not differ among treatments within the same protein source, with greater (P ≤ 0.05) values for gilts on the SP treatments than the CS treatments. However, N retained/N absorbed and N retained/N intake (%) were less (P ≤ 0.05) and PUN concentrations were greater (P ≤ 0.05) for SP treatments with cecal OTC infusion. The overall fecal biotin concentration mean was 2.6-fold greater than the overall ileal biotin concentration mean. In conclusion, no gilts in the current experiment became biotin deficient because the biotin requirements were met primarily by microbial synthesis and absorption of biotin from the distal small intestine and large intestine, with corn and soybean meal contributing endogenous biotin. Therefore, supplementation of diets for gilts entering the breeding herd with 100% of the current NRC biotin requirement for sows is adequate.

Effect of continuous v. daytime grazing on feed intake and growth of sheep grazing in a semi-arid grassland steppe

In the Inner Mongolian steppe, China, sheep generally graze during daytime and are kept in yards overnight. Hence, nutrients are not returned to the grassland, which might reduce its long-term productivity. Furthermore, the restricted grazing time may limit forage intake and thus the performance of sheep. The aim of this study was therefore to evaluate the impact of continuous 24-h grazing (CG) v. the common daytime grazing (DG) on herbage mass (HM), feed quality, feed organic matter intake (OMI) and live weight gain (LWG) of sheep in the Inner Mongolian steppe. Experiments were carried out from July to September, between 2005 and 2007 on two 2-ha plots per grazing treatment. Each month, the external faecal marker titanium dioxide (TiO2) was orally administered to six sheep per plot on 10 consecutive days. Faecal grab samples were obtained from day 6 to 10 and analysed for CP concentrations to estimate digestibility of organic matter (dOM). Faecal TiO2 concentrations were used to determine the total faecal output and hence OMI of sheep. Concomitant to faeces collection, HM and quality, as well as LWG of the animals were measured. HM and herbage quality did not differ between treatments. However, as the season progressed, concentrations of NDF, ADF and ADL increased, whereas HM and CP concentrations declined. HM and herbage quality parameters differed between years according to the annual precipitation. dOM was similar in CG (0.577) and DG (0.572) sheep, but it decreased from July (0.583) to September (0.558) and differed between years. Accordingly, the OMI of sheep was similar for both treatments across the entire grazing season and all study years. Although mean LWG differed between months and years, it was identical in CG (101.5 g/day) and DG sheep (101.8 g/day). Additional time on pasture during night does not increase feed intake or animal performance. Positive effects of a nutrient reflux on grassland productivity and herbage quality when animals remained on the plots overnight were not found, most likely because of the fact that sheep crowded together in one corner of the plots during darkness and faecal and urinary excretions were thus not evenly distributed across the entire pasture. Considering the risk of animal theft as well as the importance of sheep manure as fuel, the common penning of sheep at night seems to be an adequate management practice for pastoralists in the Inner Mongolian steppe.

Influence of wet distillers grains diets on beef cattle fecal bacterial community structure

BackgroundThe high demand for ethanol in the U.S. has generated large stocks of wet distillers grains (DG), a byproduct from the manufacture of ethanol from corn and sorghum grains. Little is known, however, about the potential influence of dietary DG on fecal microbial community structure. A better understanding of the microbial population in beef cattle feces could be an important monitoring tool to facilitate goals of improving nutrient management, increasing animal growth performance and decreasing odors and/or shedding of pathogens. Five diets consisting of a traditional diet fed to finishing beef cattle in the Southern High Plains of Texas-CON (steam-flaked corn control with 0% DG), and four concentrations of DG in the dietary dry matter; 10 C (10% corn-based DG), 5S (5% sorghum-based DG), 10S (10% sorghum DG), and 15S (15% sorghum DG) were fed to steers at the Texas Tech University Burnett Animal Center. Diets were essentially isonitrogenous with a formulated crude protein value of 13.5%.ResultsFecal grab samples were obtained from 20 steers (n = 4 per diet) and the barcoded DNA pyrosequencing method was used to generate 127,530 16S operational taxonomic units (OTUs). A total of 24 phyla were observed, distributed amongst all beef cattle on all diets, revealing considerable animal to animal variation, however only six phyla (core set) were observed in all animals regardless of dietary treatment. The average abundance and range of abundance, respectively of the core phyla were as follows: Firmicutes (61%, 19 to 83%), Bacteroidetes (28%, 11 to 63%), Proteobacteria (3%, 0.34 to 17.5%), Tenericutes (0.15%, 0.0 to 0.35%), Nitrospirae (0.11%, 0.03 to 0.22%), and Fusobacteria (0.086%, 0.017 to 0.38%). Feeding DG-based diets resulted in significant shifts in the fecal microbial community structure compared with the traditional CON. Four low abundance phyla significantly responded to dietary treatments: Synergistetes (p = 0.01), WS3 (p = 0.054), Actinobacteria (p = 0.06), and Spirochaetes (p = 0.06).ConclusionsThis is, to our knowledge, the first study using this method to survey the fecal microbiome of beef cattle fed various concentrations of wet DG. Comparison of our results with other cattle DNA sequencing studies of beef and dairy cattle feces from a variety of geographical locations and different management practices identifies a core set of three phyla shared across all cattle. These three phyla, in order of relative abundance are; Firmicutes, Bacteroidetes, and Proteobacteria. The presence of large animal-to-animal variation in cattle microbiome was noted in our study as well as by others.

Effects of subacute ruminal acidosis challenges on fermentation and endotoxins in the rumen and hindgut of dairy cows

The effects of a grain-based subacute ruminal acidosis (SARA) challenge (GBSC) and an alfalfa-pellet SARA challenge (APSC) on fermentation and endotoxins in the rumen and in the cecum, as well as on endotoxins in peripheral blood, were determined. Six nonlactating Holstein cows with cannulas in the rumen and cecum were used in the study. A 3×3 Latin square arrangement of treatments with 4-wk experimental periods was adopted. During the first 3 wk of each experimental period, all cows received a diet containing 70% forages [dry matter (DM) basis]. In wk 4 of each period, cows received 1 of the following 3 diets: the 70% forage diet fed during wk 1 to 3 (control), a diet in which 34% of the dietary DM was replaced with grain pellets made of 50% ground wheat and 50% ground barely (GBSC), or a diet in which 37% of dietary DM was replaced with pellets of ground alfalfa (APSC). Rumen pH was monitored continuously using indwelling pH probes, and rumen fluid, blood, cecal digesta, and fecal grab samples were collected immediately before feed delivery at 0900h and at 6h after feed delivery on d 3 and 5 of wk 4. The time for which rumen pH was below 5.6 was 56.4, 225.2, and 298.8min/d for the control, APSC, and GBSC treatments, respectively. Compared with the control, SARA challenges resulted in similar reductions in cecal digesta pH, which were 7.07, 6.86, and 6.79 for the control, APSC, and GBSC treatments, respectively. Compared with the control, only GBSC increased starch content in cecal digesta, which averaged 2.8, 2.6, and 7.4% of DM for the control, APSC, and GBSC, respectively. Free lipopolysaccharide endotoxin (LPS) concentration in rumen fluid increased from 10,405 endotoxin units (EU)/mL in the control treatment to 30,715 and 168,391EU/mL in APSC and GBSC, respectively. Additionally, GBSC increased the LPS concentration from 16,508 to 118,522EU/g in wet cecal digesta, and from 12,832 to 93,154EU/g in wet feces. The APSC treatment did not affect LPS concentrations in cecal digesta and feces. All concentrations of LPS in blood plasma were below the detection limit of >0.05EU/mL of the technique used. Despite the absence of LPS in blood, only GBSC increased the concentration of LPS-binding protein in blood plasma, which averaged, 8.9, 9.5, and 12.1mg/L for the control, APSC, and GBSC treatments, respectively. This suggests that GBSC caused translocation of LPS from the digestive tract but that LPS was detoxified before entering the peripheral blood circulation. The higher LPS concentration in cecal digesta in the GBSC compared with the APSC suggests a higher risk of LPS translocation in the large intestine in GBSC than in APSC.

Effect of reduced ferulate-mediated lignin/arabinoxylan cross-linking in corn silage on feed intake, digestibility, and milk production

Cross-linking of lignin to arabinoxylan by ferulates limits in vitro rumen digestibility of grass cell walls. The effect of ferulate cross-linking on dry matter intake (DMI), milk production, and in vivo digestibility was investigated in ad libitum and restricted-intake digestion trials with lambs, and in a dairy cow performance trial using the low-ferulate sfe corn mutant. Silages of 5 inbred corn lines were fed: W23, 2 W23sfe lines (M04-4 and M04-21), B73, and B73bm3. As expected, the W23sfe silages contained fewer ferulate ether cross-links and B73bm3 silage had a lower lignin concentration than the respective genetic controls. Silages were fed as the sole ingredient to 4 lambs per silage treatment. Lambs were confined to metabolism crates and fed ad libitum for a 12-d adaptation period followed by a 5-d collection period of feed refusals and feces. Immediately following the ad libitum feeding trial, silage offered was limited to 2% of body weight. After a 2-d adaptation to restricted feeding, feed refusals and feces were collected for 5 d. Seventy Holstein cows were blocked by lactation, days in milk, body weight, and milk production and assigned to total mixed ration diets based on the 5 corn silages. Diets were fed for 28 d and data were collected on weekly DMI and milk production and composition. Fecal grab samples were collected during the last week of the lactation trial for estimation of feed digestibility using acid-insoluble ash as a marker. Silage, total mixed ration, feed refusals, and fecal samples were analyzed for crude protein, starch, neutral detergent fiber (NDF), cell wall polysaccharides, and lignin. The W23sfe silages resulted in lower DMI in the ad libitum trial than the W23 silage, but DMI did not differ in the restricted trial. No differences were observed for NDF or cell wall polysaccharide digestibility by lambs with restricted feeding, but the amount of NDF digested daily increased for lambs fed the M04-21 W23sfe silage ad libitum. Lambs were less selective against NDF and lignin when offered W23sfe silages. The B73bm3 silage did not affect DMI or digestibility of cell walls at the restricted feeding level, but total daily NDF digested was greater at ad libitum intake. Intake, milk production, and cell wall digestibility were greater for cows fed diets containing W23sfe silages than for those fed W23 silage. Although milk production was greater for the B73bm3 diet, DMI and cell wall digestibility were not altered. Cows were less selective against cell wall material when fed both W23sfe and B73bm3 silages. Reduced ferulate cross-linking in sfe corn silage is a new genetic mechanism for improving milk production.

Effect of graded levels of niacin supplementation of a semipurified diet on energy and nitrogen balance, growth performance, diarrhea occurrence, and niacin metabolite excretion by growing swine1

Thirty-six crossbred barrows with an average initial age of 42 d and BW of 13.8 kg were placed in individual metabolism crates in a 35-d experiment to evaluate the supplementation of a semipurified diet with graded levels of crystalline niacin. Response criteria were energy and N balance, growth performance, occurrence of niacin deficiency diarrhea, and urinary excretion of the niacin metabolite N(1)-methyl-2-pyridone-5-carboxylamide (PYR). The basal diet met the true ileal Trp requirement of growing swine, and supplementation with 6, 10, 14, 18, 22, or 44 mg of niacin/kg made 6 treatments. Pigs were observed for scours twice daily, and pig BW and feed consumption were determined weekly. Total urine collections and fecal grab samples were made twice daily from each pig from d 28 to 35. Pigs fed the diet containing 14 mg of niacin/kg absorbed and retained more (P < 0.05) grams of N/d, had a greater N digestibility (%, P < 0.05), a greater ADFI and ADG (P < 0.10), and no diarrhea (P < 0.05) compared with pigs fed the diet containing 6 mg of niacin/kg, and pigs fed the diet containing 10 mg of niacin/kg were intermediate in ADG. There were no additional improvements in the response criteria with niacin supplementation greater than 14 mg/kg. Urinary PYR criteria (mg/L and mg/d) were greater (P < 0.001) for pigs fed the diet containing 44 mg of niacin/kg than for pigs fed the diets containing 6 to 22 mg of niacin/kg. However, urinary PYR criteria for pigs fed the diets containing 6 to 22 mg of niacin/kg did not differ from each other, indicating that PYR was not a sensitive indicator of niacin status for growing swine. Niacin treatment did not affect the percentages of N retained/N absorbed, N retained/N intake, DE, or ME. In conclusion, 14 mg of crystalline niacin/kg of semipurified diet adequate in Trp was the minimum concentration of niacin that maximized N utilization and growth performance, and prevented niacin deficiency diarrhea of growing swine in the current experiment. Because practical feed ingredients may be sources of available endogenous niacin, supplementation of practical diets with 100% of the current NRC requirement for niacin should provide adequate niacin for growing swine.

Naturally colonized beef cattle populations fed combinations of yeast culture and an ionophore in finishing diets containing dried distiller's grains with solubles had similar fecal shedding of Escherichia coli O157:H7.

Beef steers (n = 252) were used to evaluate the effects of dietary supplement on fecal shedding of Escherichia coli O157:H7. Seven pens of 9 steers (63 steers per treatment) were fed diets supplemented with or without yeast culture (YC) or monensin (MON) and their combination (YC × MON). YC and MON were offered at 2.8 g/kg and 33 mg/kg of dry matter intake, respectively. Environmental sponge samples (from each pen floor, feed bunk, and water trough) were collected on day 0. Rectal fecal grab samples were collected on days 0, 28, 56, 84, 110, and 125. Samples were collected and pooled by pen and analyzed for presumptive E. coli O157:H7 colonies, which were confirmed by a multiplex PCR assay and characterized by pulsed-field gel electrophoresis (PFGE) typing. On day 0, E. coli O157:H7 was detected in 7.0% of feed bunk samples and 14.3% of pen floor samples but in none of the water trough samples. The 71.4% prevalence of E. coli O157:H7 in fecal samples on day 0 decreased significantly (P < 0.05) over time. E. coli O157:H7 fecal shedding was not associated with dietary treatment (P > 0.05); however, in cattle fed YC and YC × MON fecal shedding was 0% by day 28. Eight Xba I PFGE subtypes were identified, and a predominant subtype and three closely related subtypes (differing by three or fewer bands) accounted for 78.7% of environmental and fecal isolates characterized. Results from this study indicate that feeding YC to cattle may numerically decrease but not eliminate fecal shedding of E. coli O157:H7 at the onset of treatment and that certain E. coli O157 subtypes found in the feedlot environment may persist in feedlot cattle.

Fecal excretion patterns and short term bias of internal and external markers in a digestion assay with cattle

The objective of this work was to evaluate the fecal excretion profile and the short term bias of the external markers chromic oxide and titanium dioxide and the internal markers indigestible dry matter (iDM), indigestible neutral detergent fiber (iNDF) and indigestible acid detergent fiber (iADF) in a digestion trial with cattle fed different diets. Fourteen F1 Red Angus × Nellore bulls averaging 12 months old and 287 kg were kept in individual stalls. The animals were fed elephant grass silage, corn silage or signal grass hay, supplemented or not with 20% of concentrate mixture. The experiment consisted of two 13-day experimental periods according to a 2 × 2 Latin square design with seven squares grouping. The animals received 10 g of chromic oxide and 10 g of titanium dioxide through esophageal sounder daily. Fecal grab samples were obtained directly from the rectum of the animals at 1:30 a.m., 3:00 a.m., 4:30 a.m., 6:00 a.m., 7:30 a.m., 9:00 a.m., 10:30 a.m., 12:00 p.m., 1:30 p.m., 3:00 p.m., 4:30 p.m., 6:00 p.m., 7:30 p.m., 9:00 p.m., 10:30 p.m. and 12:00 a.m. There was no effect of forages or concentrate levels on the dimension of excretory cycle. The fundamental period of excretory cycle was 15.06 and 18.66 hours for chromic oxide and titanium dioxide, respectively and 9.93, 9.29 and 10.55 hours for iDM, iNDF and iADF, respectively. The oscillation range was higher for external markers. Considering the characteristics of all evaluated markers together, it is recommended fecal sampling designs with at least four collections, distributed during the day or only in the diurnal period for obtaining fecal excretion estimates free of short term bias.

Effects of limit feeding corn or dried distillers grains with solubles at 2 intakes during the growing phase on the performance of feedlot cattle1

Energy density in growing diets may affect carcass quality of cattle; however, few reports have described the impact of energy source. The objectives of this research were to determine effects of source [dried distillers grains with solubles (DDGS) vs. corn] and amount (limit-fed to gain 0.9 vs. 1.4 kg of BW/d) of energy during the growing phase on feedlot performance and marbling. Angus-cross steers (144 head) were blocked by BW (average initial BW = 252 ± 36 kg), allotted within each block to 8 pens (6 steers/pen, 24 pens total), and randomly assigned to 1 of 4 feeding systems in a 2 × 2 factorial arrangement of treatments: 1) 65% DDGS fed to gain 0.9 kg of BW/d, 2) 65% DDGS fed to gain 1.4 kg of BW/d, 3) 65% corn fed to gain 0.9 kg of BW/d, and 4) 65% corn fed to gain 1.4 kg of BW/d. Fecal grab samples were collected on d 52 of the growing phase to determine digestibility of DM, ADF, NDF, ether extract (EE), and CP. After the 98-d growing phase, all steers were fed the same finishing diet. Steers were slaughtered by pen when average BW within the pen was 544, 522, and 499 kg for the large, medium, and small BW blocks, respectively. Average daily gain and DMI differed (P<0.01) by design during the growing phase. Compared with the corn-based diets, digestibilities of DM, NDF, and EE were decreased (P<0.02) when DDGS-based diets were fed during the growing phase, whereas the digestibility of N was increased (P<0.01). The ADG was greatest (P=0.02) during the finishing phase for steers fed to gain 0.9 kg of BW/d initially, but source of energy during the growing phase did not affect (P=0.24) finishing phase ADG. Steers fed to gain 0.9 kg of BW/d during the growing phase also had less backfat (P=0.08), decreased USDA yield grades (P=0.03), and greater LM area (P<0.01) than steers fed to gain 1.4 kg of BW/d. There was an interaction between energy source and amount for marbling scores (P=0.02). Steers fed corn-based diets to gain 0.9 kg of BW/d during the growing phase had the most marbling, whereas those fed to gain 0.9 kg of BW/d on DDGS had the least marbling; the remaining feeding systems were intermediate. Overall ADG and DMI were affected (P < 0.06) by both source and amount of energy fed during the growing phase. Feeding the DDGS-based diet to achieve greater ADG during the growing phase increased marbling, whereas feeding the corn-based diet to increase ADG during the growing phase decreased marbling.

Use of plant hydrocarbons as markers to estimate voluntary intake and digestibility in beef steers

Calculating dry matter intake of grazing herbivores has been an issue in determining factors such as efficiency and digestibility of forages. Two experiments were conducted using alkanes to validate analytical methods with oven-dried and freeze-dried samples, compare and contrast measured and estimated intake, and to measure digestibility using a controlled-release capsule (CRC) or supplement. In the first trial, 7 beef steers consuming either alfalfa or switchgrass hay received either a CRC or supplement in a random crossover design. Each period lasted 20 days with fecal grab samples taken over the last 10 days and total fecal collection the last 5 days. In the second trial, 12 beef cattle were fed a blended switchgrass and alfalfa hay and fed a once-daily alkane supplement. Fecal grab samples were collected the last 5 days over the 14-day experiment. In the first experiment, steers fed switchgrass hay ate less and digested a smaller proportion of total diet ( P < 0.02) than steers fed alfalfa. There was no difference ( P < 0.35) between estimated intake and measured intake. Fecal concentrations of alkanes were not affected ( P < 0.13) by drying method and no difference ( P < 0.40) was seen between fecal grab samples and total fecal collection. Either dotriacontane (C32) or hexatriacontane (C36) can be used for estimating DMD. In the second experiment, there was no difference between measured and estimated intake ( P < 0.35). There was no day effect between fecal concentrations and estimated intake. There was no difference ( P < 0.65) between fecal alkane concentrations when duplicate samples were analyzed. Alkanes can be used to estimate intake in cattle. Fecal grab samples are sufficient to reduce labor and result in adequate estimated intake and daily feeding of alkanes can be used to replace the CRC.

Technical note: Use of near-infrared reflectance spectroscopy to predict intake and digestibility in bulls and steers1

Multiple fecal samples were collected from growing Angus bulls (264 to 419 kg of BW, 3.0 to 11.4 kg/d of DMI) to predict DMI of a corn-silage-based diet. Contemporaneous digestion trials were conducted with the same diet in 12 steers in yr 1 to 3 and bulls in yr 4. Near-infrared spectra from fecal samples (n = 730 from 282 growing bulls, n = 240 from 36 steers and 12 bulls for digestion trials) were obtained from dried and ground fecal samples, and modified partial least squares regression was used to develop equations to predict DMI and DM digestibility (DMD). Although mean predicted DMI of the growing bulls (7.52 ± 0.04 kg/d or 22.4 ± 0.1 g/kg of BW) was within 2% of mean measured DMI (7.63 ± 0.06 kg/d or 22.7 ± 0.1 g/kg of BW), the mean of paired differences within samples (0.11 ± 0.04 kg/d or 0.3 ± 0.1 g/kg of BW) was greater (P < 0.01) than zero. Measured DMD (72.3 ± 0.5%) was identical (P < 0.97) to predicted DMD (72.3 ± 0.5%), and DMD for bulls in the digestion trial did not differ (P < 0.27) from DMD for steers. Prediction of intake requires incorporation of some measured values from the set of fecal samples to be predicted. Lack of similarity between spectra of fecal grab samples from the growing bulls and daily fecal collection of steers and bulls in the digestion trials in this study indicates the need for further verification before prediction of DMD with fecal grab samples.

Assessment of Diagnostic Tools for Identifying Cattle Shedding and Super-SheddingEscherichia coliO157:H7 in a Longitudinal Study of Naturally Infected Feedlot Steers in Ohio

The objectives of this study were to compare the performance of different diagnostic protocols (rectoanal mucosal swabs and immunomagnetic separation [RAMS-IMS], fecal samples and IMS [fecal-IMS], and direct plating) to determine the prevalence of Escherichia coli O157:H7 and to evaluate the pattern of E. coli O157:H7 shedding and super-shedding (defined as having a direct plating count equal to or >10(4) colony forming units of E. coli O157:H7 per gram of feces) in a longitudinal study of naturally infected feedlot steers. RAMS and fecal grab samples were obtained at 14-day intervals from 168 Angus-cross beef steers over a period of 22 weeks. Fecal samples were assessed by direct plating and IMS, whereas RAMS were tested only by enrichment followed by IMS to recover E. coli O157:H7. The period prevalence for shedding was high (62%) among feedlot steers and super-shedding was higher (23%) than anticipated. Although direct plating was the least sensitive method to detect E. coli O157:H7-positive samples, over 20% of high bacterial load samples were not detected by RAMS-IMS and/or fecal-IMS. The sensitivity of RAMS-IMS, fecal-IMS, and direct plating protocols was estimated using simple and multilevel mixed-effects logistic regression models, in which the dependent variable was the dichotomous results of each test and gold standard (i.e., parallel interpretation of the three protocols)-positive individuals were included as an independent variable along with other factors such as dietary supplements, time of sampling, and being exposed to a super-shedding pen-mate. The associations between these factors and the sensitivity of the diagnostic protocols were not statistically significant. In conclusion, differences in the reported impact of diet and probiotics on the shedding of E. coli O157:H7 in previous studies using RAMS-IMS or fecal-IMS were unlikely due to their impact on test performance.

A Randomized Controlled Trial to Assess the Impact of Dietary Energy Sources, Feed Supplements, and the Presence of Super-Shedders on the Detection of Escherichia coli O157:H7 in Feedlot Cattle Using Different Diagnostic Procedures

Alteration of the gastro-intestinal tract through manipulation of cattle diets has been proposed as a preharvest control measure to reduce fecal shedding of Escherichia coli O157:H7. The objective of this study was to examine the effects of the energy source's moisture content (high moisture corn and dry whole-shelled corn), two natural feed supplements (Saccaromyces cerevisiae boulardii CNCM 1079-Levucell and Aspergillus oryzae-Amaferm), and two levels of vitamin A (2200 IU/kg and no supplementation) on the fecal excretion of E. coli O157:H7 in naturally infected cattle. One hundred sixty-eight Angus-cross beef steers were randomly allocated to 24 pens, and each pen was assigned 1 of 12 dietary treatments in a randomized complete block design. E. coli O157:H7 was detected by rectoanal mucosal swab (RAMS) and fecal grab samples using immunomagnetic separation (IMS) and standard microbiological techniques. On the basis of multivariable multilevel logistic regression models, we found a statistically significant (p < 0.05) increase in the prevalence of E. coli O157:H7 in animals fed dry whole-shelled corn in models based on fecal-IMS, and this effect was increased if a super-shedding animal (shedding > 10(4) colony forming units of E. coli O157:H7 per gram of feces) was present in the pen at the time of testing relative to animals fed high moisture corn and nonexposed to super-shedders. However, in similar models based on RAMS-IMS testing, the effect of corn type on the prevalence of E. coli O157:H7 varied with the type of feed additive used. Being exposed to a super-shedding pen-mate also increased the odds of being positive to E. coli O157:H7 in the RAMS-IMS models. These models demonstrate that the impact of different supplements may vary with the diagnostic test used, and that further research into the biological significance of differences between RAMS- and fecal-IMS test results is warranted.

Oral delivery systems for encapsulated bacteriophages targeted at Escherichia coli O157:H7 in feedlot cattle.

Bacteriophages are natural predators of bacteria and may mitigate Escherichia coli O157:H7 in cattle and their environment. As bacteriophages targeted to E. coli O157:H7 (phages) lose activity at low pH, protection from gastric acidity may enhance efficacy of orally administered phages. Polymer encapsulation of four phages, wV8, rV5, wV7, and wV11, and exposure to pH 3.0 for 20 min resulted in an average 13.6% recovery of phages after release from encapsulation at pH 7.2. In contrast, untreated phages under similar conditions had a complete loss of activity. Steers (n = 24) received 10(11) CFU of naladixic acid-resistant E. coli O157:H7 on day 0 and were housed in six pens of four steers. Two pens were control (naladixic acid-resistant E. coli O157:H7 only), and the remaining pens received polymer-encapsulated phages (Ephage) on days -1, 1, 3, 6, and 8. Two pens received Ephage orally in gelatin capsules (bolus; 10(10) PFU per steer per day), and the remaining two pens received Ephage top-dressed on their feed (feed; estimated 10(11) PFU per steer per day). Shedding of E. coli O157:H7 was monitored for 10 weeks by collecting fecal grab and hide swab samples. Acceptable activity of mixed phages at delivery to steers was found for bolus and feed, averaging 1.82 and 1.13 x 10(9) PFU/g, respectively. However, Ephage did not reduce shedding of naladixic acid-resistant E. coli O157:H7, although duration of shedding was reduced by 14 days (P < 0.1) in bolus-fed steers as compared with control steers. Two successful systems for delivery of Ephage were developed, but a better understanding of phage-E. coli O157:H7 ecology is required to make phage therapy a viable strategy for mitigation of this organism in feedlot cattle.

Effect of dry- versus wet-autoclaving of spray-dried egg albumen compared with casein as protein sources on apparent nitrogen and energy balance, plasma urea nitrogen and glucose concentrations, and growth performance of neonatal swine1

Forty crossbred neonatal pigs with an average initial age of 4 d and BW of 2.16 kg were used in a 28-d experiment to evaluate the nutritional effects of autoclaving a commercial sugar-free, spray-dried egg albumen (EA) compared with casein. Basal diet protein sources were lactic acid casein and EA. Two more dietary treatments were made by replacing the EA with dry-autoclaved EA (DAEA) or wet-autoclaved EA (WAEA, EA and water mixed in a 1.0:1.2 ratio before autoclaving). The DAEA and WAEA were autoclaved at 121 degrees C and 1.75 kg/cm(2) pressure for 30 min, and WAEA was oven-dried after autoclaving. Analyzed trypsin inhibitor units/mg of EA, DAEA, and WAEA were 535.0, 9.0, and 6.5, respectively. Pigs were fed the diets in gruel form to appetite in individual metabolism cages every 2 h during the experiment. Blood samples were taken on d 7, 14, and 21, and total urine and fecal grab-samples were collected from d 14 to 21 of the experiment. Response criteria were N and energy balance, plasma urea N (PUN) and glucose concentrations, and growth performance. The WAEA was a higher quality protein source for neonatal pigs than DAEA. Pigs fed the diet containing WAEA absorbed and retained more (P < 0.05) grams of N/d, had higher (P < 0.05) percentages of N and energy that were absorbed and retained/intake, had lower (P < 0.05) concentrations of PUN overall, and had higher (P < 0.05) ADG and G:F than pigs fed the diet containing DAEA. Most response criteria of pigs fed the diets containing DAEA or EA were not different, although pigs fed the diet containing DAEA had lower (P < 0.05) overall PUN concentrations, and pigs fed the diet containing EA had higher (P < 0.05) percentages of energy absorbed and retained/intake, and higher ADG and G:F than pigs fed the diet containing DAEA. Growth performance was not different for pigs fed the diets containing WAEA or casein. However, pigs fed the diet containing casein excreted less (P < 0.05) fecal N, retained more (P < 0/05) grams of N/d, had higher percentages of N absorbed and retained/intake, and had lower (P < 0.05) PUN concentrations overall than pigs fed the diet containing WAEA. In conclusion, WAEA was a higher quality protein source for neonatal pigs than DAEA or EA, whereas lactic casein was a higher quality protein source for neonatal pigs than EA, DAEA, or WAEA.

Escherichia coli O157:H7 Strains That Persist in Feedlot Cattle Are Genetically Related and Demonstrate an Enhanced Ability To Adhere to Intestinal Epithelial Cells

A longitudinal study was conducted to investigate the nature of Escherichia coli O157:H7 colonization of feedlot cattle over the final 100 to 110 days of finishing. Rectal fecal grab samples were collected from an initial sample population of 788 steers every 20 to 22 days and microbiologically analyzed to detect E. coli O157:H7. The identities of presumptive colonies were confirmed using a multiplex PCR assay that screened for gene fragments unique to E. coli O157:H7 (rfbE and fliC(h7)) and other key virulence genes (eae, stx(1), and stx(2)). Animals were classified as having persistent shedding (PS), transient shedding (TS), or nonshedding (NS) status if they consecutively shed the same E. coli O157:H7 genotype (based on the multiplex PCR profile), exhibited variable E. coli O157 shedding, or never shed morphologically typical E. coli O157, respectively. Overall, 1.0% and 1.4% of steers were classified as PS and NS animals, respectively. Characterization of 132 E. coli O157:H7 isolates from PS and TS animals by pulsed-field gel electrophoresis (PFGE) typing yielded 32 unique PFGE types. One predominant PFGE type accounted for 53% of all isolates characterized and persisted in cattle throughout the study. Isolates belonging to this predominant and persistent PFGE type demonstrated an enhanced (P < 0.0001) ability to adhere to Caco-2 human intestinal epithelial cells compared to isolates belonging to less common PFGE types but exhibited equal virulence expression. Interestingly, the attachment efficacy decreased as the genetic divergence from the predominant and persistent subtype increased. Our data support the hypothesis that certain E. coli O157:H7 strains persist in feedlot cattle, which may be partially explained by an enhanced ability to colonize the intestinal epithelium.

Prevalence and Impact of Bacteriophages on the Presence of Escherichia coli O157:H7 in Feedlot Cattle and Their Environment

The relationship between endemic bacteriophages infecting E. coli O157:H7 (referred to as "phage") and levels of shedding of E. coli O157:H7 by cattle was investigated in two commercial feedlots in southern Alberta, Canada. Between May and November 2007, 10 pens of cattle were monitored by collection of pooled fecal pats, water with sediment from troughs, manure slurry from the pen floor, and rectal fecal samples from individual animals (20 per pen) at two separate times. Bacteriophages infecting E. coli O157:H7 were detected more frequently (P<0.001) after 18 to 20 h enrichment than by initial screening and were recovered in 239 of 855 samples (26.5% of 411 pooled fecal pats, 23.8% of 320 fecal grab samples, 21.8% of 87 water trough samples, and 94.6% of 37 pen floor slurry samples). Overall, prevalence of phage was highest (P<0.001) in slurry. Recovery of phage from pooled fecal pats was highest (P<0.05) in May. Overall recovery did not differ (P>0.10) between fecal grab samples and pooled fecal pats. A higher prevalence of phage in fecal pats or water trough samples was associated (P<0.01) with reduced prevalence of E. coli O157:H7 in rectal fecal samples. There was a weak but significant negative correlation between isolation of phage and E. coli O157:H7 in fecal grab samples (r= -0.11; P<0.05). These data demonstrate that the prevalence of phage fluctuates in a manner similar to that described for E. coli O157:H7. Phage were more prevalent in manure slurry than other environmental sources. The likelihood of fecal shedding of E. coli O157:H7 was reduced if cattle in the pen harbored phage.

The effects of live yeast supplementation to dairy cows during the hot season on production, feed efficiency, and digestibility

The current study examined the effects of live yeast (LY) supplementation to dairy cows during the summer season on milk production, feed efficiency and ration digestibility. Forty-two dairy cows (14 primiparous and 28 multiparous) were fed either a control lactating diet or supplemented with 1g of LY (Saccharomyces cerevisiae, Biosaf, Lesaffre) per 4kg of dry matter consumed. The LY amounts were adjusted twice a week. Four rumen samples were taken from 30 cows in 2-h periods and ammonia concentrations were determined. Fecal grab samples from 30 cows were collected during 3 consecutive days, to determine the apparent digestibility of diets. The daily dry matter intake in the LY group was 2.5% greater compared with the control group (24.7 and 24.1kg, respectively). The daily average milk yield of the LY group was greater by 1.5kg (4.1%) compared with the control group (37.8 vs. 36.3kg, respectively). There were no significant differences in the milk fat and protein percentages, but fat yield was greater in the LY group than in the control. The fat-corrected milk 4% was 2.0kg (6.1%) greater in the LY group than in the control group (34.8 vs. 32.8kg, respectively). The efficiency of using dry matter to produce 4% fat-corrected milk was 3.7% greater in the LY group compared with the control group. The ruminal ammonia concentrations after feeding were greater in the control group than in the LY group (151.9 vs. 126.1 mg/l, respectively). No differences were observed among groups in the total tract apparent digestibility of dry matter and other diet components. The pH values in the rumen that were determined in a companion trial using 4 fistulated cows tended to be higher in cows that were supplemented with LY than in the control (6.67 vs. 6.54, respectively). It may be concluded that LY supplementation to dairy cows during the hot season improved the rumen environment in a way that increased the dry matter intake and in consequence enhanced the productivity and efficiency.

Baixa dosagem de óxido de cromo para estimativa da produção fecal em bovinos

Foi avaliado o uso de baixa dosagem de óxido de cromo (Cr2O3) incorporado em um alimento concentrado para estimativa da produção fecal (PF) em bovinos. Para tanto, foram conduzidos quatro ensaios de digestibilidade in vivo utilizando quatro novilhos com peso vivo médio de 214±31kg, recebendo ad libitum no cocho azevém anual (Lolium multiflorum Lam.) cortado verde. Aproximadamente 200g de ração contendo 5g kg-1 do indicador foi fornecida diariamente durante 12 dias, sendo feito a coleta de dados e amostras nos últimos cinco dias de cada período. A PF foi medida com o uso de sacolas de coleção total ou estimada com base na concentração do indicador em amostras fecais coletadas duas vezes ao dia (8 e 16h) ou a intervalos de duas horas entre 8 e 22h. A concentração média de cromo nas amostras coletadas às 8 e 16h (445mg kg-1) foi semelhante à média geral de todos os horários (447mg kg-1). O grau de recuperação fecal (GR) do indicador aumentou linearmente, de aproximadamente 40% para em torno de 80%, com o aumento da concentração fecal de cromo até esta última atingir um valor em torno de 250mg kg-1 de MO. Quando a concentração de cromo nas fezes foi superior a este valor o GR se manteve relativamente constante, em média 76%. A produção fecal foi superestimada pelo indicador em até 35% quando não corrigido para o GR. Quando corrigido para a recuperação fecal, as estimativas de produção fecal de MO estimadas foram similares às observadas. A excreção fecal de bovinos alimentados com azevém pode ser acuradamente estimada com o uso de baixa dosagem de óxido de cromo incorporado em uma ração peletizada concomitante à coleta de duas amostragens diárias de fezes.