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Hydrolyzed feather meal (HFM) is a feedstuff high in rumen undegraded protein (RUP) that can be used as an effective source of metabolizable protein for dairy cattle. Because the production process may vary, the rumen degradability and intestinal digestibility of HFM may also vary. Additionally, some processes may incorporate additional blood into the final product to result in feather meal with poultry blood. To determine the rumen degradability and intestinal digestibility of these products, several laboratory assays can be used; the common assays are the mobile bag (MOB), modified three-step (MTS), and Ross (ROS) assays. Although all 3 assays determine RUP digestibility, they vary in whether they are performed in situ, in vitro, or both. The objective of this study was to evaluate the ruminal degradability and intestinal digestibility of HFM originating from processes that differ in their inclusion of blood, and to compare the MOB, MTS, and ROS assays. Ten samples of HFM, which were identified by the suppliers as HFM with little blood (n = 5) and with more blood (n = 5), were spot-sampled, collected from 10 production plants across the United States, and subjected to all 3 assays. Assay type had an effect on RUP, total-tract crude protein (CP) digestibility, and the amount of RUP digested. A significant effect was observed on RDP and RUP concentrations for blood inclusion; no effect was detected for total-tract CP digestibility. We found no difference in RUP digestibility for assay or blood inclusion. There was also no interaction of the effect of assay or blood inclusion. Results suggest that even though there are differences in chemical composition in HFM associated with the inclusion of blood, such as ash and crude fat, few if any differences are observed in intestinal digestion of protein. Although the assays varied in their estimates of rumen undegraded protein, MOB and MTS yielded the most similar values. However, all 3 assays resulted in similar estimates of RUP digestibility.

Maintenance energy is the energy required to conserve the state of an animal when no work is completed. Dietary energy must be supplied to meet maintenance requirements before milk can be produced. The objectives of the current experiment were to quantify the maintenance energy requirement of Jersey cows when lactating or dry. Energetic measures were collected on 8 Jersey cows and evaluated across 3 physiological phases and nutritional planes: lactation, dry cows fed at maintenance, and fasted dry cows. Through total collection of feces and urine as well as using headbox-style indirect calorimeters, energy balance and heat production data were measured across all phases. Lactation data were collected across four 28-d periods. Data for cows fed at maintenance were collected after 14 d and fasting heat production was measured during the last 24 h of a 96-h fast. Net energy for maintenance (NEM) requirements, and the efficiency of converting metabolizable energy (ME) into net energy were compared between lactating and dry (maintenance or fasting phase) cows. Heat production of dry cows fed at maintenance, which represents ME for maintenance, was 0.146 ± 0.0087 Mcal per unit of metabolic body weight (BW0.75, MBW). Fasting heat production, which represents NEM, was 0.102 ± 0.0071 Mcal/MBW. Energy balance was calculated as tissue energy plus milk energy. When estimated via regressing energy balance on ME intake, NEM was not different between dry and lactating cows (0.120 ± 0.32 vs. 0.103 ± 0.0052 Mcal/MBW). However, the slope of the regression of energy balance on ME intake was greater for dry compared with lactating cows (0.714 ± 0.046 vs. 0.685 ± 0.010) when evaluated with a fixed intercept. This suggests that dry cows were more efficient at converting ME into net energy and that the efficiency of utilizing ME for maintenance may be greater than for lactation. Our measurements of NEM and the slope of ME on energy balance were greater than the value used by the National Research Council (2001), which are 0.080 Mcal/MBW for NEM and approximately 0.64 for the slope. Results of this study suggest that NEM and the efficiency of converting ME into NEM of modern lactating Jersey cows are similar to recent measurements on modern Holstein cows and greater than previous measurements.

Measurement of urinary energy (UE) excretion is essential to determine metabolizable energy (ME) supply. Our objectives were to evaluate the accuracy of using urinary N (UN) or C (UC) to estimate UE and ultimately improve the accuracy of estimating ME. Individual animal data (n = 433) were used from 11 studies with Jersey cows at the University of Nebraska–Lincoln, where samples were analyzed after drying (n = 299) or on an as-is basis (n = 134). Dried samples resulted in greater estimated error variance compared with as-is samples, and thus only as-is samples were used for final models. The as-is data set included a range (min to max) in dry matter intake (11.6–24.6 kg/d), N intake (282–642 g/d), UE excretion (1,390–3,160 kcal/d), UN excretion (85–220 g/d or 20.6–59.5% of N intake), and UC excretion (130–273 g/d). As indicated by a bias in residuals between observed and predicted ME as dietary crude protein (CP; range of 14.9–19.1%) increased, the National Research Council dairy model did not accurately predict ME of diets, as dietary CP varied. The relationship between UE (kcal/d) and UN (g/d) excretion was linear and had an intercept of 880 ± 140 kcal. Because an intercept of 880 is biologically unlikely, the intercept was forced through 0, resulting in linear and quadratic relationships. The regressions of UE (kcal/d) on UN (g/d) excretion were UE = 14.6 ± 0.32 × UN, and UE = 20.9 ± 1.0 × UN − 0.0357 ± 0.0056 × UN2. In the quadratic regression, UE increased, but at a diminishing rate as UN excretion increased. As UC increased, UE linearly and quadratically increased. However, error variance was greater for regression with UC compared with UN as explanatory variables (8.42 vs. 7.42% of mean UE). The use of the quadratic regression between UN and UE excretion to predict ME resulted in a slope bias in ME predictions as dietary CP increased. The linear regression between UE and UN excretion removed slope bias between predicted ME and CP, and thus may be more appropriate for predicting UE across a wider range of dietary CP. Using equations to predict UE from UN should improve our ability to predict diet ME in Jersey cows compared with calculating ME directly from digestible energy.

A simulation study was conducted to examine accuracy of estimating daily O2 consumption, CO2 and CH4 emissions, and heat production (HP) using a spot sampling technique and to determine optimal spot sampling frequency (FQ). Data were obtained from 3 experiments where daily O2 consumption, emissions of CO2 and CH4, and HP were measured using indirect calorimetry (respiration chamber or headbox system). Experiment 1 used 8 beef heifers (ad libitum feeding; gaseous exchanges measured every 30 min over 3 d in respiration chambers); Experiment 2 used 56 lactating Holstein-Friesian cows (restricted feeding; gaseous exchanges measured every 12 min over 3 d in respiration chambers); Experiment 3 used 12 lactating Jersey cows (ad libitum feeding; gaseous exchanges measured every hour for 1 d using headbox style chambers). Within experiment, averages of all measurements (FQALL) and averages of measurements selected at time points with 12, 8, 6, or 4 spot sampling FQ (i.e., sampling every 2, 3, 4, and 6 h in a 24-h cycle, respectively; FQ12, FQ8, FQ6, and FQ4, respectively) were compared. Within study a mixed model was used to compare gaseous exchanges and HP among FQALL, FQ12, FQ8, FQ6, and FQ4, and an interaction of dietary treatment by FQ was examined. A regression model was used to evaluate accuracy of spot sampling within study [i.e., FQALL (observed) vs. FQ12, FQ8, FQ6, or FQ4 (estimated)]. No interaction of diet by FQ was observed for any variables except for CH4 production in experiment 1. No FQ effect was observed for gaseous exchanges and HP except in experiment 2 where CO2 production was less (5,411 vs. 5,563 L/d) for FQ4 compared with FQALL, FQ12, and FQ8. A regression analysis between FQALL and each FQ within study showed that slopes and intercepts became farther from 1 and 0, respectively, for almost all variables as FQ decreased. Most variables for FQ12 and FQ8 had root mean square prediction error (RMSPE) less than 10% of the mean and concordance correlation coefficient (CCC) greater than 0.80, and RMSPE increased and CCC decreased as FQ decreased. When a regression analysis was conducted with combined data from the 3 experiments (mixed model with study as a random effect), results agreed with those from the analysis for the individual studies. Prediction errors increased and CCC decreased as FQ decreased. Generally, all the estimates from FQ12, FQ8, FQ6, and FQ4 had RMSPE less than 10% of the means and CCC greater than 0.90 except for FQ6 and FQ4 for O2 consumption and CH4 production. In conclusion, the spot sampling simulation with 3 indirect calorimetry experiments indicated that FQ of at least 8 samples (every 3 h in a 24-h cycle) was required to estimate daily O2 consumption, CO2 and CH4 production, and HP and to detect changes in those in response to dietary treatments. This sampling FQ may be considered when using techniques that measure spot gas exchanges such as the GreenFeed and face mask systems.

A study using indirect calorimetry and 12 lactating multiparous Jersey cows (53 ± 23 d in milk at the beginning of the experiment; mean ± standard deviation) was conducted to evaluate the utilization of energy in cattle consuming diets containing increasing hydrolyzed feather meal (HFM). A triplicated 4 × 4 Latin square design with 35-d periods (28-d adaption and 4-d collections) was used to compare 4 different dietary treatments. Treatments contained (DM basis) HFM at 0% (0HFM), 3.3% (3.3HFM), 6.7% (6.7MFM), and 10.0% (10HFM). Diets were formulated such that HFM replaced blood meal and nonenzymatically browned soybean meal. With increasing HFM, linear increases were observed for dietary NEL content (1.61, 1.64, 1.69, and 1.70 ± 0.042 Mcal/kg of DM for 0HFM, 3.3HFM, 6.7MFM, and 10HFM, respectively), and the efficiency of converting ME to NEL (0.708, 0.711, 0.717, and 0.719). Apparent total-tract digestibility of CP linearly decreased with increasing HFM (63.4, 61.1, 59.9, and 58.6 ± 1.46% for 0HFM, 3.3HFM, 6.7MFM, and 10HFM, respectively), whereas long-chain fatty acid digestibility increased with increasing HFM (77.2, 77.7, 78.5, and 80.6 ± 1.30%). With increased inclusion of HFM, fecal N excretion increased (199, 230, 239, 237 ± 12.1 g/d for 0HFM, 3.3HFM, 6.7MFM, and 10HFM, respectively), whereas urinary N excretion decreased (166, 151, 155, and 119 ± 14.8 g/d). Increasing the concentration of HFM resulted in a quadratic effect on DMI (19.6, 20.2, 20.3, and 19.1 ± 0.79 kg/d for 0HFM, 3.3HFM, 6.7MFM, and 10HFM, respectively) and milk yield (31.7, 32.0, 31.9, and 29.7 ± 1.32 kg/d). Increasing HFM linearly decreased the milk protein concentration (3.34, 3.29, 3.23, and 3.23 ± 0.158 for 0HFM, 3.3HFM, 6.7MFM, and 10HFM, respectively) and yield (1.05, 1.05, 1.02, and 0.96 ± 0.040 kg). The inclusion of HFM did not affect energy-correct milk yield (average of 39.3 ± 1.54). Results of this study suggest that HFM can increase dietary NEL content compared with blood meal and nonenzymatically browned soybean meal and maintained energy-corrected milk yield; however, feeding HFM at greater than 6.7% of diet DM decreased DMI, and protein availability may have been reduced with increased HFM, leading to a linear decrease in milk protein concentration and yield.

An experiment was conducted to validate and optimize the procedure of spot urine sampling with urinary creatinine as a marker to estimate urine outputs of dairy cows. Twelve lactating cows were used in a randomized complete block design. Cows were grouped and randomly assigned to 2 experimental diets: a corn silage-based diet and an alfalfa silage-based diet with supplemental potassium. The experiment lasted for 21 d and total collection (TC) of urine was conducted for the last 3 d. Twelve spot samples of urine from individual cows were collected over a 3-d period during TC to represent every 2-h sampling in a 24-h cycle. Creatinine excretion rate (mg/kg of body weight per d) was variable among cows from 16.7 to 34.5 with an average of 27.3. Creatinine concentrations of spot samples within cow were averaged to simulate urine samples obtained from various spot sampling frequencies (equally spaced 12, 6, 4, and 2 time points starting at feeding: 12TP, 6TP, 4TP, and 2TP, respectively). Large diurnal variation of urinary creatinine concentration was observed within cow. Creatinine concentration was greater (75 vs. 65 mg/dL) for 12TP compared with TC, resulting in underestimating (29.8 vs. 32.6 kg/d) urine outputs. When compared among 12TP, 6TP, 4TP, and 2TP, creatinine concentrations were different and urine outputs tended to be different for 2TP compared with 12TP, 6TP, and 4TP. In addition, despite underestimation of urine output, a regression analysis indicated strong linear relationships between 12TP, 6TP, or 4TP and TC, suggesting that this technique can successfully identify the differences in urine outputs altered by dietary treatments. However, 4TP failed to detect statistical differences in urine outputs between a corn silage-based diet and the alfalfa silage-based diet with supplemental potassium, indicating that a spot urine sampling frequency of at least 6 was required to identify dietary effects on urine outputs. According to the pattern of diurnal changes in urinary creatinine concentration, a spot sample at about 10 h after feeding may have potential to obtain a urine sample that is more representative (i.e., creatinine concentration) of TC urine compared with urine from multiple sampling frequencies. Overall, urinary creatinine as a marker with spot sampling of urine underestimated urine output. However, 12TP and 6TP were successful in identifying changes in urine outputs by dietary treatments.

The effects of dietary fatty acid (FA) and starch content as well as supplemental digestible Lys (sdLys) on production, energy utilization, and N utilization were evaluated. Each factor was fed at 5 different amounts, and factor limits were as follows: 3.0 to 6.2% of dry matter (DM) for FA; 20.2 to 31.3% of DM for starch, and 0 to 17.8 g/d of sdLys. Dietary FA and starch were increased by replacing soyhulls with supplemental fat and corn grain, respectively, and sdLys increased with rumen-protected Lys. Fifteen unique treatments were fed to 25 Jersey cows (mean ± SD; 80 ± 14 d in milk) across 3 blocks in a partially balanced incomplete block design. Each block consisted of 4 periods of 28 d, where the final 4 d were used to determine milk production and composition, feed intake, energy utilization (via total collection and headbox-style indirect calorimetry), and N utilization (via total collection). Response surface models were used to evaluate treatment responses. Increasing dietary FA decreased DM intake and milk protein yield. When dietary starch was less than 24%, milk protein concentration increased with increasing sdLys, but when dietary starch was greater than 26% milk protein concentration decreased with increasing sdLys. Digestibility of FA increased when dietary FA increased from 3.0 to 4.2% and decreased as FA increased beyond 4.2%. Although neutral detergent fiber digestibility decreased as dietary starch increased, energy digestibility increased. As dietary FA increased, metabolizable energy (ME) content quadratically increased. Supply of ME increased as dietary FA increased from 3.0 to 4.2% and decreased as FA increased beyond 4.2%. Increasing dietary FA and starch decreased CH4 production and urinary energy. Increasing dietary starch increased the efficiency of utilizing dietary N for milk N. Increasing sdLys quadratically decreased N balance as sdLys increased from 0 to 8 g/d and increased N balance as sdLys increased from 8 to 18 g/d. Increasing dietary FA can increase ME content, however, at high dietary FA, decreased DM intake and FA digestibility resulted in a plateau in ME content and a decrease in ME supply. Our results demonstrate that sdLys supply is important for milk protein when dietary starch is low, and some Lys may be preferentially used for muscle protein synthesis at the expense of milk protein when sdLys is high.

Hydrolyzed feather meal (HFM) is a feed that is high in rumen undegradable protein; however, it is low in Lys compared with other high rumen undegradable protein sources. Additionally, processing methods differ by facility, which affects AA composition and protein digestibility. The objective of this study was to use lactating dairy cows to determine the effects of feeding 2 sources of HFM that differed by the amount of blood they contained and also to study the effects of supplementing rumen-protected (RP) Lys when these sources of HFM are fed. In this study, 12 multiparous Jersey cows were enrolled in a triplicated 4 × 4 Latin square with 4 periods 28 d in length. Cows were fed 2 total mixed rations that differed by source of HFM. The HFM was included at 4.5% of the diet dry matter, and one source was produced with the addition of poultry blood. Cows were randomly assigned to 1 of 4 treatment sequences. Treatments were as follows: HFM without added blood and no RP-Lys, HFM with added blood and no RP-Lys, HFM without blood and with RP-Lys (22 g of digestible Lys), and HFM with added blood and RP-Lys. The source of HFM containing blood tended to increase dry matter intake (18.3 vs. 17.3 ± 0.72 kg/d), and increased milk yield (20.5 vs. 18.4 ± 1.31 kg/d) and protein yield (0.788 vs. 0.694 ± 0.040 kg/d). The inclusion of RP-Lys did not affect milk or protein yield. In cows fed HFM containing blood, plasma concentration of Lys (82.1 vs. 70.8 ± 4.06 μM) and His (27.8 vs. 17.9 ± 3.15 μM) was higher. The addition of RP-Lys had no effect on the concentration of either plasma Lys or His. Gross energy intake tended to increase for HFM containing more blood (81.4 vs. 77.3 ± 3.29 Mcal/d); however, no difference was observed for intake of digestible energy (52.0 ± 2.20 Mcal/d) or metabolizable energy (46.4 ± 2.02 Mcal/d). Similar to dry matter intake, N intake increased with the inclusion of HFM containing blood, but crude protein digestibility decreased (61.6 vs. 66.0%). Results of this study highlight that source of HFM can be a factor that affects milk production and that this in part is due to differences in the profile of AA. Additionally, the observation that plasma His and milk protein increased with the consumption of HFM containing more blood suggests that His may have played a role in increasing milk and milk protein yield.

The physical form of feeds can influence dairy cow chewing behavior, rumen characteristics, and ruminal passage rate. Changing particle size of feeds is usually done through grinding or chopping forages, but pelleting feed ingredients also changes particle size. Our objective was to determine if pelleted dried distillers grains and solubles (DDGS) affected the feeding value for lactating dairy cattle. Seven lactating Jersey cows that were each fitted with a ruminal cannula averaging (± standard deviation) 56 ± 10.3 d in milk and 462 ± 75.3 kg were used in a crossover design. The treatments contained 15% DDGS in either meal or pelleted form with 45% or 55% forage on a dry matter basis. The forages were alfalfa hay, corn silage, and wheat straw. The factorial treatment arrangement was meal DDGS and low forage (mDDGS-LF), pelleted DDGS and low forage (pDDGS-LF), meal DDGS and high forage (mDDGS-HF), and pelleted DDGS and high forage (pDDGS-HF). Dry matter intake and energy-corrected milk were both unaffected by treatment averaging 19.8 ± 2.10 kg/d and 33.9 ± 1.02 kg/d, respectively. Fat yield was unaffected averaging 1.7 ± 0.13 kg/d, but protein yield was affected by the interaction of forage and DDGS. Protein yield was similar for both low forage treatments but was increased by when pDDGS was fed in the high forage treatment (1.05 vs. 0.99 ± 0.035 kg/d). When forage concentration was increased, starch digestibility increased by 1.9 percentage units, crude protein digestibility tended to increase 1.1 percentage units, and residual organic matter digestibility decreased 3.4 percentage units. Pelleting DDGS increased digestibility of neutral detergent fiber (NDF) digestibility (49.2 vs. 47.5 ± 1.85%) and gross energy (68.2 vs. 67.1 ± 1.18%). Increasing forage increased ruminal pH (5.85 to 5.94 ± 0.052). Passage rate slowed from 2.84 to 2.65 ± 0.205 %/h when feeding HF compared with LF. Rumination time increased from 417 to 454 ± 49.4 min with increasing forage concentration but was unaffected by the form of DDGS or the interaction of forage and DDGS. Eating time increased with pDDGS (235 vs. 209 ± 19.8 min), which may be a result of increased feed sorting behavior. Pelleting DDGS increased preference for particles retained on the 8-mm sieve and decreased preference for particles on the 1.18-mm sieve and in the pan (1.18 mm). Results confirm that increasing forage concentration increases ruminal pH, rumination time, and slows passage rate, but contrary to our hypothesis increasing forage concentration did not increase NDF digestibility. Results also suggest that pelleted DDGS do not appear to affect milk production, ruminal characteristics, or passage rate, but pelleted DDGS may increase sorting behavior of lactating Jersey cows and increase NDF and gross energy digestibility.

Heat production (HP) represents a major energy cost in lactating dairy cows. Better understanding of factors that affect HP will improve our understanding of energy metabolism. Our objective was to derive models to explain variation in HP of lactating Jersey cows. Individual animal-period data from 9 studies (n = 293) were used. The data set included cows with a wide range (min to max) in days in milk (44–410) and milk yield (7.8–43.0 kg/d). Diets included corn silage as the predominate forage source, but diets varied (min to max on DM basis) in crude protein (CP; 15.2–19.5%), neutral detergent fiber (NDF; 35.5–43.0%), starch (16.2–31.1%), and crude fat (2.2 to 6.4%) contents. Average HP was (mean ± standard deviation) 22.1 ± 2.86 Mcal/d, or 28.1 ± 3.70% of gross energy intake. Eight models were fit to explain variation in HP: (1) dry matter intake (DMI; INT); (2) milk fat, protein, and lactose yield (MILKCOMP); (3) INT and milk yield (INT+MY); (4) INT and MILKCOMP/DMI (INT+MILKCOMP); (5) mass of digested NDF, CP, and starch (DIG); (6) INT and digested energy (INT+DE); (7) INT and NDF, CP, and starch digestibility (INT+DIG); or (8) INT+MILKCOMP model plus urinary N excretion (INT+MILKCOMP+UN). For all HP models, metabolic body weight was included. All models were derived via a backward elimination approach and included the random effects of study, cow, and period within block within study. The INT models adequately explained variation in HP with a nonrandom effect–adjusted concordance correlation coefficient of 0.84. Similar adjusted concordance correlation coefficients (0.79–0.85) were observed for other HP models. The HP associated with milk protein yield and supply of digestible protein was greater than other milk production and nutrient digestibility variables. The HP associated with urinary N excretion was 5.32. Overall, HP can be adequately predicted from metabolic body weight and DMI. Milk component yield, nutrient digestibility, or urinary N excretion explained similar variation as DMI. Coefficients for milk protein and protein digestion suggest that digestion and metabolism of protein and synthesis of milk protein contribute substantially to HP of a dairy cow.

The objective of this study was to determine the effects of high-starch or high-fat diets formulated to be isoenergetic on energy and N partitioning and utilization of energy. Twelve multiparous Jersey cows (mean ± standard deviation; 192 ± 11 d in milk; 467 ± 47 kg) in a crossover design with 28-d periods (24-d adaptation and 4-d collection) were used to compare 2 treatment diets. Treatments were high starch (HS; 30.8% starch, 31.8% neutral detergent fiber, and 1.9% fatty acids) or high fat (HF; 16.8% starch, 41.7% neutral detergent fiber, and 4.1% fatty acids). Diets were formulated to have net energy for lactation (NEL) content of 1.55 Mcal/kg of dry matter according to the National Research Council (2001) dairy model. Nutrient composition was varied primarily by replacing corn grain in HS with a rumen-inert fat source and cottonseed hulls in HF. Gross energy content was lower for HS (4.43 vs. 4.54 ± 0.01 Mcal/kg of dry matter), whereas digestible (2.93 vs. 2.74 ± 0.035 Mcal/kg of dry matter) and metabolizable energy (2.60 vs. 2.41 ± 0.030 Mcal/kg of dry matter), and NEL (1.83 vs. 1.67 ± 0.036 Mcal/kg of dry matter) content were all greater than for HF. Tissue energy deposited as body fat tended to be greater for HS (4.70 vs. 2.14 ± 1.01 Mcal/d). For N partitioning, HS increased milk N secretion (141 vs. 131 ± 10.5 g/d) and decreased urinary N excretion (123 vs. 150 ± 6.4 g/d). Compared with HF, HS increased apparent total-tract digestibility of dry matter (66.7 vs. 61.7 ± 1.06%), organic matter (68.5 vs. 63.2 ± 0.98%), energy (66.0 vs. 60.4 ± 0.92%), and 18-carbon fatty acids (67.9 vs. 61.2 ± 1.60%). However, apparent total-tract digestibility of starch decreased for HS from 97.0 to 94.5 ± 0.48%. Compared with HF, HS tended to increase milk yield (19.7 vs. 18.9 ± 1.38 kg/d), milk protein content (4.03 vs. 3.93 ± 0.10%), milk protein yield (0.791 vs. 0.740 ± 0.050 kg/d), and milk lactose yield (0.897 vs. 0.864 ± 0.067 kg/d). In addition, HS decreased milk fat content (5.93 vs. 6.37 ± 0.15%) but did not affect milk fat yield (average of 1.19 ± 0.09 kg/d) or energy-corrected milk yield (average of 27.2 ± 1.99 kg/d). Results of the current study suggest that the HS diet had a greater metabolizable energy and NEL content, increased partitioning of N toward milk secretion and away from urinary excretion, and may have increased partitioning of energy toward tissue energy deposited as fat.

The objective of the experiment was to examine effects of a diet containing a high concentration (28.8% dry matter basis) of corn distillers grain with solubles on manure characteristics and NH3 and H2S emissions from dairy cow manure. Eighteen cows were blocked by parity and days in milk, and cows in each block were assigned to the following treatments: the control diet (CON) or CON with distillers grains with solubles at 28.8% (dry matter basis) replacing mainly soybean meal (DG). The experiment was conducted for 11 wk, and feces and urine from individual cows were collected over 3 d in wk 11 (a total of 8 spot samples per cow). Fecal or urine samples were composited by cow, and the composite feces and urine were analyzed for indigestible neutral detergent fiber and creatinine concentration, respectively, for individual cows to estimate total fecal and urine outputs. Immediately before the manure incubation, composited feces and urine were sampled to determine manure characteristics. Manure was reconstituted according to daily fecal and urine excretion estimated for individual cows. Individual manures were incubated using a continuous air flux multichamber system over 10 d to measure NH3 and H2S emissions. All data from 18 manures were analyzed using the Mixed procedure of SAS (SAS Institute Inc., Cary, NC). The ratio of feces to urine and the contents of manure total and volatile solids were not different among treatments. Urine from DG had lower pH and DG manure had lower N content and greater S content compared with CON. During the 10-d incubation, NH3 emission was considerably less for DG versus CON. The emission of H2S over 10 d for DG was greater compared with that for CON. After the incubation, manure pH and N and S concentrations were greater for DG versus CON. In conclusion, manure from cows fed a high-DG diet decreased urinary N contribution to manure N and lowered urine pH, which were the factors that caused the decrease in NH3 emission from DG manure. However, the DG diet increased dietary S concentration and increased S excretion in urine and feces. This increased H2S emission from DG manure during the 10-d manure incubation.

Nitrogen concentrations in feeds, feces, milk, and urine samples were measured using 2 analytical methods following different drying procedures. Ten samples of corn silage, alfalfa silage, and concentrates collected from 2017 to 2018 at Krauss Dairy Research Center, The Ohio State University (Wooster), were used. A 4-d total collection digestion trial provided fecal samples from 10 cows (1 sample/cow), and another 10 cows were used to collect milk samples (1 sample/cow) and spot urine samples (1 sample/cow). Spot urine samples were acidified immediately to pH

Background Peritoneal metastasis from biliary carcinoma (PMC) is associated with poor prognosis when treated with chemotherapy. Objective To evaluate the impact on survival of cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC), and compare with conventional palliative chemotherapy for patients with PMC. Material and methods A prospective multicenter international database was retrospectively searched to identify all patients with PMC treated with a potentially curative CRS/HIPEC (CRS/HIPEC group). The overall survival (OS) was compared to patients with PMC treated with palliative chemotherapy (systemic chemotherapy group). Survival was analyzed using Kaplan-Meier method and compared with Log-Rank test. Results Between 1995 and 2015, 34 patients were included in the surgical group, and compared to 21 in the systemic chemotherapy group. In the surgical group, median peritoneal cancer index was 9 (range 3–26), macroscopically complete resection was obtained for 25 patients (73%). There was more gallbladder localization in the surgical group compared to the chemotherapy group (35% vs. 18%, p = 0.001). Median OS was 21.4 and 9.3 months for surgical and chemotherapy group, respectively (p=0.007). Three-year overall survival was 30% and 10% for surgical and chemotherapy group, respectively. Conclusion Treatment with CRS and HIPEC for biliary carcinoma with peritoneal metastasis is feasible and may provide survival benefit when compared to palliative chemotherapy.

Indirect methods of spot sampling with intrinsic markers to estimate fecal output and nutrient digestibility often have been used in dairy nutrition research as alternatives to total collection of feces (TC) because of labor and expense. However, fecal output and nutrient digestibility estimated from the indirect method must be accurate regardless of altering dietary conditions. This experiment was designed to validate the accuracy of using indigestible neutral detergent fiber (iNDF) or acid-insoluble ash (AIA) as intrinsic markers to estimate fecal outputs and nutrient digestibility compared with TC and to determine the optimal number of spot sampling events to accurately determine fecal output and then nutrient excretion. The experiment used 12 multiparous lactating Holstein cows in a randomized complete block design. Cows were blocked by days in milk and milk yield and randomly assigned to 1 of 2 diets: a diet containing about 49% corn silage on a dry matter basis and a diet containing about 48% alfalfa silage with high by-product (soyhulls) and supplemental K. During the final 3 d of 21-d periods, TC was performed, and 12 spot samples were collected for the same 3 d to represent every 2 h in a 24-h cycle. Fecal outputs and nutrient digestibility of dry matter, organic matter, or nitrogen estimated with iNDF or AIA as an intrinsic marker were compared with TC. Overall, fecal outputs and digestibility estimated with iNDF were similar to that estimated with TC, whereas AIA overestimated fecal output by 44 to 61% and underestimated nutrient digestibilities by 16 to 32%. However, potential differences in statistical inference of dietary effects between iNDF and TC were found. Data from individual spot samples were aggregated to represent spot sampling frequencies of 12 (SP12), 6 (SP6), 4 (SP4), or 2 (SP2) evenly spaced events starting at feeding time. Compared with TC, SP12 produced similar fecal content of iNDF, organic matter, and nitrogen, but fecal AIA content was greater. Furthermore, compared with SP12, SP6 produced similar fecal content of all nutrients, whereas marker and nutrient concentrations in SP4 and SP2 were different. In this experiment, iNDF was a better fecal marker than AIA, and a spot sampling frequency of at least 6 events was necessary. However, interpretation of dietary effects could be confounded when iNDF was used to estimate fecal outputs.

This study investigated effects of high inclusion of reduced-fat corn distillers grains with solubles (RFDG) with or without monensin on utilization and excretion of dietary N, P, and S. The experiment was conducted for 11 wk (2-wk diet adaptation, 9-wk experimental period of data collection) with 36 Holstein cows in a randomized complete block design. Cows were blocked by parity, days in milk, and milk yield and assigned to the following diets: (1) a control diet (CON); (2) CON with RFDG included at 28.8% (dry matter basis) by replacing soybean meal, soyhulls, and supplemental fat and phosphorus (DG); and (3) DG with monensin (Rumensin; Elanco Animal Health, Greenfield, IN) supplemented at a rate of 20 mg/kg of DM offered (DGMon). Contrasts were used to compare CON versus DG and DG versus DGMon. Inclusion of RFDG at 28.8% of dietary DM replacing mainly soybean meal did not change crude protein content (17.6% on a DM basis) but decreased rumen-degradable protein and increased rumen-undegradable protein. In addition, the DG diets increased P (0.48 vs. 0.36%) and S concentrations (0.41 vs. 0.21%; DM basis) compared with the CON diet. As a result, DG versus CON decreased plasma and milk urea N concentrations and urinary N excretion. However, the increase in P concentration when feeding the DG versus CON diet to lactating cows increased P intake, plasma P concentration, and urinary and fecal P excretion without affecting milk P secretion. Intake of S was greater for cows fed the DG versus CON diet, resulting in greater plasma total S and sulfate concentration and urinary and fecal S excretion. However, milk S secretion was not affected by DG compared with CON. Monensin supplementation to the DG diet did not affect N intake, utilization, and excretion except that apparent N digestibility was lower compared with DG. In addition, feeding the DGMon diet did not affect P and S utilization and excretion compared with DG. The study suggests that inclusion of high RFDG in a ration by replacing mainly soybean meal altered N, P, and S utilization and excretion, but monensin supplementation to a high-RFDG diet, overall, had minimal effects on N, P, and S utilization and excretion in lactating dairy cows.

This study investigated the effects of continuous feeding of high inclusion of reduced-fat corn distillers grains with solubles with and without monensin on dry matter intake (DMI), production, milk fatty acid profile, and plasma AA profile in lactating cows. The experiment was conducted for 12 wk (1-wk covariate, 2-wk diet adaptation, and 9-wk experimental period of data collection) with 36 Holstein cows in a randomized complete block design. Cows were blocked by parity, days in milk, and milk yield and assigned to the following diets: (1) control (CON), (2) CON with reduced-fat corn distillers grains with solubles included at 28.8% (dry matter basis) replacing soybean meal, soyhulls, and supplemental fat (DG), and (3) DG with monensin (Rumensin; Elanco Animal Health, Greenfield, IN) supplemented at a rate of 20 mg/kg of DM offered (DGMon). Orthogonal contrasts were used to compare CON versus DG and DGMon and to compare DG versus DGMon. Milk yield was not affected (40.3 vs. 40.8 kg/d) by DG and DGMon compared with CON. However, for DG and DGMon compared with CON, decreased DMI (24.9 vs. 26.4 kg/d), milk fat yield (1.12 vs. 1.55 kg/d), milk protein yield (1.24 vs. 1.32 kg/d), and energy-corrected milk yield (37.7 vs. 43.5 kg/d) were observed. Feeding DGMon compared with DG did not affect DMI (24.4 vs. 25.4 kg/d) and milk yield (39.2 vs. 41.3 kg/d) but decreased milk fat yield (1.08 vs. 1.23 kg/d), milk protein yield (1.20 vs. 1.28 kg/d), and energy-corrected milk yield (36.0 vs. 39.4 kg/d). Interactions between treatment and week for DMI, milk fat yield, and energy-corrected milk indicate that production responses to DG and DGMon versus CON were decreased over the experimental period. Cows fed DG and DGMon had increased milk fat concentration of trans-10,cis-12 18:2, trans-10 18:1, and long-chain (>16C) and polyunsaturated fatty acids and decreased short-chain (

Opinions of those in the cattle, swine, and sheep slaughtering and rendering sectors were characterized regarding aspects of the National Animal Identification System. Across plants of all size categories, operators of 51, 46, 65, and 22% of cattle, swine, and sheep slaughtering and rendering plants, respectively, said they would now, voluntarily, submit animal termination records (ATR). By size category, however, too few operators of large cattle and swine slaughtering and rendering plants are willing to do so. There are preconditions that might make slaughtering and rendering plant operators more willing to voluntarily submit ATR; 79 to 88% would do so if radio frequency identification devices (cattle and sheep) and slap-tattoo or premises identification numbers (swine) were made mandatory. Only 22, 29, 29, and 14% of cattle, swine, and sheep slaughtering and rendering plant operators, respectively, would voluntarily pay a private database provider a fee to protect ATR data from the Freedom of Information Act, and 53, 25, 31, and 57%, respectively, of plant operators would—if ATR submission were made mandatory—send their ATR data through a private database provider if the fees were paid by the government. Support for mandatory animal identification (which would include submission of ATR) across small-, medium-, and large-sized plant categories was 47 to 80, 25 to 43, 25 to 100, and 100% for cattle, swine, and sheep slaughtering and rendering plant operators, respectively, if industry had to pay for it, but increased to 65 to 90, 64 to 75, 50 to 100, and 100% if the government would pay for it.

The threat of a livestock disease outbreak or other animal health events in North America is real. However, predicting both the timing and severity of an outbreak can be extremely difficult. Animal identification and traceability programs can help limit the spread of disease. The overall objective of this review is to evaluate and compare animal identification and traceability systems in North America. Mandated animal identification programs, which exist for Canadian cattle and sheep and Mexican cattle, are designed to control and eradicate trade-limiting diseases and to maintain or gain access to international markets. In contrast, the United States has chosen to implement the National Animal Identification System as a voluntary program for cattle, sheep, and swine. However, the US sheep industry has operated with a mandatory National Scrapie Eradication Program since 2001, and the US pork industry has independently implemented a mandatory swine premises registry, which targeted 100% compliance by December 31, 2007, and a mandatory swine identification program targeting full compliance by December 31, 2008. Likewise, the Canadian National Hog Traceability and Identification System will become a mandatory program in 2008. It is recognized that a country’s ability to respond to an animal disease outbreak is greatly enhanced with the implementation of a national animal identification program.

Animal diseases and related food safety issues, such as bovine spongiform encephalopathy, have become concerns to many people in the last decade. With approximately one billion domesticated swine in the world, traceability is becoming increasingly important throughout the world as a way to control disease outbreaks before they have devastating effects on a country’s livestock and related livestock industries. The objective of this review is to discuss swine identification and traceability systems outside North America. Some countries, such as those in the European Union, have mandatory traceability programs that require that pigs be traced from birth to slaughter and that imported products to adhere to the same standard. Countries with mandatory swine traceability programs have systems in place that can document animal movement records (premises changes), as well as animal birth and termination records. Many countries that have voluntary programs, such as Australia and Brazil, are currently trying to develop comprehensive traceability programs for swine and pork.

Animal identification by means of marking animals’ bodies was first recorded 3,800 yr ago in the Code of Hammurabi, and throughout history, valuable animals such as horses have been identified to prevent thievery all over the world. Today, the reasons for identification of livestock include production management, control of disease outbreaks, establishment of ownership, requirements for export, and consumer demands. Additionally, there are many methods of animal identification and traceability available today including ear tags, tattooing, branding, electronic methods that implement radio frequency identification technologies (such as rumen boluses, ear tags, and injectable transponders), and biometric methods (such as retinal scanning, nose prints, and DNA). The objective of this review is to demonstrate the implementation of bovine animal identification and traceability systems in selected countries outside of North America (i.e., United States, Canada, and Mexico) for the purpose of creating a knowledge base whereby an effective North American bovine animal identification and traceability system may be created and implemented. This review will discuss regulatory requirements of animal identification and traceability in selected countries.

Purpose: Determining serum total prostate specific antigen (PSA) has proved to be a valuable diagnostic aid for detecting prostatic carcinoma, although the lack of specificity has limited its usefulness. Studies indicate that the use of percent free PSA would improve specificity while maintaining sensitivity. Since complexed PSA represents the major proportion of measurable PSA in serum, we determined whether it represents a single test alternative to the use of percent free PSA for the early detection of prostate cancer. Materials and Methods: Archival serum was obtained from 385 men with no evidence of malignancy on biopsy and 272 with biopsy confirmed prostate cancer. We determined the concentration and proportion of total, complexed and free PSA. Results: Receiver operating characteristics analysis using total PSA results from all samples (range 0.32 to 117 ng./ml.) indicated that the areas under the curve for complexed PSA alone as well as the free-to-total and complexed-to-total PSA ratios were similar and significantly greater than those for total PSA alone. Within the range of 85% to 95% sensitivity receiver operating characteristics analysis revealed that the specificity of complexed PSA was higher than that of total PSA and equivalent to that of the free-to-total PSA ratio. We noted a similar improvement in specificity in the 4 to 10 ng./ml. total PSA range. Using published cutoff values for complexed, total and percent free PSA when total PSA was in the 4 to 10 ng./ml. range the sensitivity and specificity of complexed and percent free PSA were similar. Within the 4 to 10 ng./ml. total PSA range the population of patients with no evidence of malignancy and complexed PSA below the upper limit was different with respect to total PSA from that with no evidence of malignancy and free PSA greater than 25%. Conclusions: The measurement of complexed PSA represents an alternative to the use of percent free PSA, although the patient populations identified by the 2 tests are different.

A prospective study was conducted to quantify the individual animal and maternal factors that affected morbidity and mortality of calves in Colorado beef herds. The study subjects were all calves born in ten participating herds during the 1990 and 1991 calving seasons. All 3666 calves born during the study period were individually identified at birth, and subsequent disease events were recorded by the producers. The disease outcomes of interest and their observed incidence rates were: perinatal (birth to 12 h) mortality, 2.5%; general neonatal (12 h to 45 days) mortality, 2.2%; general neonatal morbidity, 4.4%; neonatal diarrhea, 1.1%; neonatal respiratory disease, 1.0%; mothering problems/weak calves, 1.2%; and neonatal enterotoxemia/sudden death, 1.4%. Data analysis utilized multiple logistic regression. All analyses were adjusted for herd. Perinatal mortalily was greater ( P P P =0.06). General neonatal mortality was higher ( P

The effect of morbidity during early life on the weaning weight of calves in Colorado beef herds was investigated as part of a prospective longitudinal observational study. A total of 2609 calves born in nine participating herds during the 1990 and 1991 calving seasons were monitored for disease events and subsequently weighed at weaning as a measure of performance. Morbidity outcomes of interest and their observed incidence rates were: general neonatal (to 45 days) morbidity, 2.6%; neonatal diarrhea, 1.0%; neonatal respiratory disease, 1.0%; and mothering problems/weak calves, 0.4%. Mean calf weaning weight was 244±46 kg. Weaning weight data were adjusted by multiple regression for the effects of the herd, year, age of the calf, age of the dam, calf sex, frame size of the dam, and twin birth. General morbidity during the neonatal period resulted in a 15.9 kg reduction (P

Study objectives: Many patients with suspected acute coronary syndrome (ACS) are admitted from the emergency department (ED) because of the insensitivity of presentation ECGs and troponin levels. Ischemia-modified albumin (IMA) is a novel cardiac biomarker of acute cardiac ischemia. The US Food and Drug Administration has approved the use of a normal IMA concentration in low-risk (per history/physical examination, ECG, and troponin) patients with chest pain syndrome as an indication for safe ED discharge. Methods: We reviewed all studies identified by computerized literature and obtained proprietary data from the assay manufacturer that evaluated IMA as an ACS “rule-out” in the ED setting. For trials to be included, patients were required to present to the ED within 3 hours of symptom onset. For each study, the appropriate IMA diagnostic cutoff was defined by receiver operating characteristic curve analysis or a normal range study. Studies eligible for inclusion were required to document discharge diagnosis with respect to cardiac ischemia or pertinent clinical outcomes for a minimum of 30 days after the index presentation. Results: There were 8 studies meeting eligibility criteria. Three trials evaluated all nontraumatic chest pain presentations; 4 studies evaluated low- to moderate-risk patients with nondiagnostic ECG and a negative troponin at presentation; and 1 study evaluated patients initially triaged as high risk and requiring a cardiology consult. IMA in conjunction with troponin and ECG at presentation had a high negative predictive value for ACS at the index visit and during the follow-up period: 84% in a high-risk population and 91% to 100% in the low-risk population. Conclusion: IMA has a high negative predictive value for short-term adverse outcomes in a preselected low- to moderate-risk population of patients with suspected ACS and may be used to facilitate contraction of the risk stratification process for chest pain syndrome in the ED.

We describe a case of simultaneous tuberculous and pyogenic infection of the pancreas in a 58-year-old Asian man who presented with a pyrexia of unknown origin. There was no evidence of disseminated tuberculosis or immuno-deficiency. The diagnosis was confirmed by ultrasound-guided percutaneous pancreatic aspiration and subsequent progress assessed by sequential computed tomography (CT) and ultrasonography. Complete recovery was achieved on anti-tuberculous chemotherapy without surgical intervention.

Scrotal circumference (SC) measurements were taken of 921 American Brahman bulls located on two ranches in southcentral Texas. Measurements were taken at various seasons of the year over an 11-yr time span. Body condition scores, as reflected by fat cover, and a subjective estimate of overall nutritional status were 4 or better, based upon a 1 to 9 scoring system, with 5 representing an animal with an average amount of body fat. Associated body weights were not measured and semen evaluations were not performed. Ages were calculated from exact birthdates and rounded to the nearest whole month. Ages from 8 mo to more than 36 mo were reported. SC measurements ranged from 16.0 cm to 42.5 cm. The relationship of SC and age was positively correlated (r=0.74) (P

Forty Santa Gertrudis bulls were used to examine relationships among scrotal circumference, seminal quality, libido, and fertility [assessed as the percent pregnant of estrous females (PE rate) and the percent pregnant of females mated (PM rate)]. These bulls were selected from 220 two year old bulls to represent variations in scrotal circumference and seminal quality. Each of the 40 bulls were exposed to 100 cyclic Santa Gertrudis heifers for a 4-day (96 hr) breeding period. The number of estrous females available to each bull varied from 12 to 27. A breeding soundness examination (BSE) was conducted on each bull approximately 45 days prior to the 4-day breeding period and immediately after the 4-day breeding period. The three components of the BSE scroe (scrotal circumference, spermatozoal abnormalities and spermatozoal motility) were not significantly correlated with PE rate or PM rate at either evaluation. There was no significant correlation between PM rate and scrotal circumference; however, each of the 4 bulls having a scrotal circumference less than 30 cm had a PM rate below 31%. Relationships between seminal quality and PM rate were unclear and differed between the two evaluations. There was a trend for bulls having poor seminal quality at the first evaluation to improve by the second evaluation. Consequently, fluctuations in seminal quality between evaluations is one possible explanation for low correlations between seminal parameters and PM rate. Libido (number mated/number in estrus x 100) was positively correlated (r = 0.44) with PE rate. Using a stepwise regression procedure, the independent variables accounting for the most variation in PE rate (dependent variable) included libido, secondary spermatozoal abnormalities, and BSE score (r(2) = 0.44). Results of this study indicate that current methods of fertility evaluation did not accurately predict the fertility of individual Santa Gertrudis bulls as measured by PE rate and PM rate during a 4 day breeding period.

The vomeronasal organ (VNO) is an accessory olfactory system which in many vertebrates seems to be involved in reproductive behavior, particularly in permitting males to detect estrus in conspecific females. We postulated that EEG-like field potentials could be recorded from the VNO because the organ has a sensory epithelium that can discharge in response to odorants, and because VNO receptor cells are structurally similar to cells in the olfactory mucosa that produce field potentials (the “electro-olfactogram”). We examined this postulate in male cattle by surgically implanting recording/perfusion cannulae into the ducts leading to each VNO. Both bipolar (between VNOs) and reference recording revealed continuous spontaneous voltage fluctuations that were similar to an EEG, except that amplitudes were larger and frequencies slower. Simultaneous recording of the EEG and VNO, using the same nasal reference electrode revealed that neither signal was “contaminated” by voltages from the other. Perfusions of one VNO with whole urine, urine condensate, or aqueous reconstituted urine extracts, whether from females in estrus or in anestrus, produced massive transient electrical responses from both VNOs. Perfusion with penicillin or local anesthetics caused more sustained large electrical responses from both VNOs, except that activity became quiescent about 15 min after local anesthesia. These results suggest that it is feasible to use direct recording of VNO responses to stimulation to investigate the behavioral physiology of the VNO. The cellular origin of these potentials is not self-evident, inasmuch as there are several probable sources: VNO receptor cells, cells of origin of the nervus terminalis, trigeminal afferents, vasomotor fibers, vascular erectile tissue, and fluid motion in the VNO. The experimental approach that is presented here could provide an assay system for exploring comparative functions and the behavioral significance of the various cell types that are associated with the mammalian VNO.

The Chironomidae are often numerically abundant in collections of aquatic invertebrates yet their specific identification is generally omitted in surveillance studies. This paper uses data collected from the R. Wye catchment to explore the influence of different levels of taxonomic penetration of the Chironomidae on community analyses often used in biological survey. The implications of the temporal distribution of the Chironomidae to surveillance sampling strategy is also considered.

ABSTRACT Self-bored pressuremeter (SBP) tests are an important part of site investigations for gravity base structure (GBS) foundations. This applies in particular when such structures are expected to experience significant lateral loads, as most arctic structures are. The SBP data is used for both serviceability state analysis and for providing fundamental information on undisturbed soil behavior. SBP testing has been carried out offshore for Gulf Canada Resources Inc. (GCRI) during the past two years (1982-1983 arctic open water seasons). Testing has been carried out in a variety of soil conditions across the GCRI land interest on the Canadian continental shelf in the Beaufort Sea. The paper outlines the equipment used, operational successes and difficulties, and presents productivity data. Techniques and equipment for improving productivity are outlined. Experience with SBP operations has shown that the SBP is no more difficult to use offshore than the electric static cone penetrometer (CPT). One result in particular of the SBP testing stands out; the in-situ geostatic stress ratio (Ko) has been found to be not a unique function of past overburden hi story. This result is discussed in the geological context. It appears prudent to regard as an independent variable for engineering purposes. INTRODUCTION An estimated 12 percent of the worlds untapped hydrocarbon reserves occur with in the Canadian Offshore regions (Beaufort Sea, Labrador Coast, Scotian Shelf and Arctic Islands). Gulf Canada Resources Inc. (GCRI) has significant interest in all these areas and is a major operator in the Beaufort Sea. The Beaufort Sea is characterized by being frozen and ice covered for about 9 months each year. This ice cover presents the principal environmental constraint in offshore operations because of the large forces it may exert on stationary structures [1]. Artifical islands have proved successful in resisting large, horizontal ice forces in shallow waters. However, the average water depth of the GCRI land interest is about 35 m (115 ft). Artifical islands would be very expensive in such water depths so caisson technology has been developed as a technique for gaining the performance of islands without their cost. The GCRI system is called the Mobile Arctic Caisson (MAC). The MAC is a hybrid soil-steel structure and the principles of its design and operation have been presented elsewhere [2], [3]. A prerequisite to deployment of the MAC is a site investigation paying particularly attention to the foundation response under inclined and dynamic load vectors. GCRI has carried out site investigations at various sites within its land interest during 1981, 1982 and 1983. However, it was found during 1981 that conventional laboratory tests gave scattered and ambiguous results at some sites.