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1 Department of Animal Science, University of Arkansas, Fayetteville 72701
2 Department of Animal Science, University of Arkansas Cooperative Extension Service, Little Rock 72203
Corresponding author: E. B. Kegley; e-mail: ekegley{at}uark.edu.
| ABSTRACT |
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1-acid glycoprotein, immunoglobulin G concentrations, and the neutrophil:lymphocyte ratio were not affected by bedding type. On d 0, coliform counts were greatest in rice hulls. After use, coliform counts were greatest in long wheat straw (week by bedding material interaction). On d 42, the concentration of ammonia at 10 cm above the bedding was lowest for long wheat straw. Growth performance of calves bedded for 42 d with 5 bedding types did not differ; however, the number of antibiotic treatments given for scours was greatest on granite fines and sand; coliform counts in the bedding were highest in rice hulls before use and in long wheat straw after 42 d of use.
Key Words: calf bedding material growth performance health bacterial count
Abbreviation key: AGP =
1-acid glycoprotein, N:L = neutrophil:lymphocyte ratio
| INTRODUCTION |
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Hunter and Houpt (1989) studied bedding preference in ponies and concluded that they preferred areas of pens bedded with wood shavings rather than bare concrete. Lactating cows preferred to use free-stalls bedded with sand or sawdust compared with mattresses (Tucker et al., 2003). However, temperature and humidity may affect the animals behavior, quality of bedding materials, and bacteria in the bedding types (Hogan et al., 1989; Bey et al., 2002). In summer, pigs prefer to lie on concrete floors, but in winter they prefer to lie on straw bedding (Fraser, 1985). Lactating cows preferred free-stalls bedded with sand as compared with mattresses in summer; however, cows used stalls bedded with mattresses and sawdust in the winter (Thoreson et al., 2000). Long wheat straw has been used traditionally for calf bedding, but it is expensive or unavailable in many areas. Other reasonably priced materials available in Central Arkansas are granite fines, rice hulls, and wood shavings.
In spite of the importance of bedding material type to keep calves clean, dry, comfortable, and healthy, there is limited information concerning how bedding material type affects calf growth performance and health. The objectives of this study were to determine the effects of 5 types of bedding material on growth, physiological stress, and behavior of calves and to monitor costs and physical characteristics of the material used.
| MATERIALS AND METHODS |
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Within 6 h of birth, 60 (58 Holstein, 1 Brown Swiss x Holstein, and 1 Guernsey x Holstein) calves were weighed and vaccinated intranasally with 2 mL of a modified live bovine rhinotracheitis and parainfluenza3 virus vaccine (TSV-2; Pfizer Animal Health, Exton, PA) and orally with 3 mL of a modified live bovine rotavirus and coronavirus vaccine (Calf-Guard; Pfizer Animal Health). Calves were injected intramuscularly with 1 mL of iron hydrogenated dextran (containing 100 mg of Fe; VEDCO, Inc., St. Joseph, MO), 1 mL of vitamin A (500,000 IU) and D (75,000 IU; VEDCO, Inc.), and 6 mL of selenium and vitamin E (2.19 mg sodium selenite/mL [1 mg selenium/mL] and 50 mg [68 USP units] of vitamin E/mL; BO-SE; Schering-Plough Animal Health Corp., Union, NJ). Each calf was identified with an ear tag, and its navel cord was dipped with 10% tincture of iodine (VEDCO, Inc.). No colostrum from the dams was fed to calves. Calves were fed 350 g of a commercial colostrum supplement (Colostrum Plus Bovine IgG-Escherichia coli Antibody; Jorgensen Laboratories, Inc., Loveland, CO) one time only within 6 h of birth by plastic esophageal feeder. Calves were reweighed at 1 d of age to establish an average initial BW. At 1 d of age, calves were randomly assigned to individual pens on 1 of 5 types of bedding materials (12 calves per treatment). Calves began the study over a 4-wk period (3 calves per treatment per week). The week that calves started was used as a block in the statistical models.
Calves were offered 2 L of medicated milk replacer (Vigortone Ag Products, Inc., Cedar Rapids, IA) mixed with Optimil concentrate calf supplement-MR type B medicated feed (Manna Pro Corp, St. Louis, MO) twice daily at 0700 and 1530 h with nipple bottles throughout this study. Medicated milk replacer was formulated to contain oxytetracycline at 220 mg and neomycin sulfate at 440 mg/kg of milk replacer powder. Optimil concentrate was formulated to contain oxytetracycline-HCl at 7.04 g and neomycin at 14.08 g/kg of Optimil powder. After 760 g of Optimil was mixed with 22.7 kg of the milk replacer, 227 g of this mixture was used the make the 2 L offered.
Commercial calf starter (Caldwell Milling Co; Rosebud, AR) was offered once daily in the morning in an open plastic bucket beginning on d 3. Calves were offered 0.12 kg/d of starter in wk 1, 0.24 kg/d in wk 2, and were increased to ad libitum in wk 3 until the weaning date after wk 6. Each morning any orts were removed, weighed, and discarded, and fresh starter was offered. Starter contained a coccidiostat (49.5 mg/ kg as fed; Deccox; Alpharma Animal Health Division, Lowell, AR). Representative samples of feed were taken every 14 d for analysis. Commercial medicated milk replacer and calf starter were analyzed for DM (AOAC, 1990), CP (nitrogen by combustion; Elemental Analyzers for the determination of nitrogen; Elementar Americas, Inc., Mt. Laurel, NJ), and fat (AOAC 920.39C, 1990). Calf starter and milk replacer samples were digested in duplicate by wet ash methodology using concentrated trace metal grade nitric acid in 50-mL conical, trace metal free, polypropylene centrifuge tubes with a heated digestion block (ModBlock, CPI International, Santa Rosa, CA). After digestion, samples were brought to 50 mL volume with a 0.1% cesium chloride solution. Concentrations of Ca were determined by atomic absorption spectroscopy (Perkin-Elmer, model 5000, Norwalk, CT). Concentrations of P were determined with a spectrophotometric procedure (AOAC, 1990) modified for use in microtiter plates. Starter and milk replacer intakes were recorded daily. Calf starter refusals were measured daily in the morning. Milk refusal was measured twice daily. Water was available at all times in an open plastic bucket. Calves were weighed initially and at 1, 2, 4, and 6 wk of age (an average of weights on 2 consecutive d was used for the final BW).
Calves were housed until weaning at 6 wk without removing any bedding materials from the pens. However, at 2-wk intervals, a bedding score was determined for each pen, then bedding materials that had been forced out around the pen were removed and placed back into that pen before adding new bedding material, if required by bedding score. The bedding score was rated on a scale of 1 to 5 as follows: 1 = dry and clean, 2 = 20 to 40% of surface dirty or wet, 3 = 40 to 60% of surface dirty or wet, 4 = 60 to 80% surface dirty or wet, and 5 = >80% of surface dirty or wet. Approximately 1.3 cm (depth) of additional bedding was added over the previous bedding when the bedding score was >3, or if less than 50% of bedding material remained in the pen, or if the bedding material became packed and a large hole developed in the surface. Fresh bedding material was added into the middle of pens or into any hole, and the weight of material added was recorded.
Calf Health Data
Calf health and cleanliness scores were recorded from 1 d of age until the weaning date. Calves were observed daily after the morning feeding. The fecal fluidity score developed by Larson et al. (1977) was used, in which 1 = normal fluidity, 2 = spreads slightly, 3 = moderately spreads, and 4 = watery (severe scours). When fecal materials were not available for scoring because of mixing with bedding materials, calves were assigned a missing value. When the fluidity score was >2, 4 mL of ceftiofur hydrochloride (Excenel; Pharmacia & Upjohn Company, Kalamazoo, MI) was given by intramuscular injection daily until fluidity score was
2. Scour days were defined as days when calves had a fluidity score >2 and received antibiotic.
Calf cleanliness score was estimated daily by a modification of the Pharmacia Animal Health Hygiene Scorecard (Pharmacia & Upjohn Co.) for milking dairy cows using a scale of 1 = calf is clean (not dirty around thighs or body), only manure at lower ends of legs, 2 = tail head region and back of calf are soiled with manure, 3 = tail head region, thighs or legs are soiled with manure, and 4 = thighs, legs, and tail head region are soiled with manure.
Blood Samples
Blood was collected (approximately 4 h after the morning milk replacer feeding) by jugular venipuncture into plain glass vacuum tubes at 1 d and at 3 and 6 wk of age. Blood was centrifuged and serum frozen (20°C) until analyzed for IgG, cortisol, and
1-acid glycoprotein (AGP). Serum concentrations of IgG (VMRD, Pullman, WA) and AGP (Cardiotech Service, Inc., Louisville, KY) were determined by commercially available radial immunodiffusion kits. Cortisol concentrations were determined by 125I radioimmunoassay (Coat-a-Count; Diagnostic Products Corp., Los Angeles, CA).
Blood was also collected in vacuum tubes containing EDTA. Blood smear slides prepared with a rapid 3-step staining kit were used to determine white blood cell differential counts (neutrophils, monocytes, lymphocytes, basophils, and eosinophils; Criterion Sciences Richard-Allan, Kalamazoo, MI), and the neutrophil to lymphocyte ratio (N:L) was calculated.
Behavioral Data
Calf behavior was recorded once weekly by walking through the barn, at a distance from the pen of at least 2.1 m, every 15 min from 0900 until 0100 h the next day (16 h). Behavior was recorded for each of the following activities by a modification of the methods of Chua et al. (2002) and Hunter and Houpt (1989) in which 1 = lying (calfs body contacted bedding and ground), 2 = standing (calf was inactive in an upright position), 3 = eating (calfs head was in plastic feed bucket), 4 = drinking (calfs head was in plastic water bucket), 5 = self-grooming (calf was licking itself), 6 = investigating environment (calf was moving or jumping around the pen), and 7 = contacting pen (calf was licking sides of the pen and/or eating bedding materials). Percentage time spent in each activity was calculated for each week.
Bedding Samples
Bedding samples were collected to measure DM and absorbency on d 0, along with pH, gram-negative bacteria counts, and coliform counts on d 0 and at 14-d intervals throughout the study. Samples on d 14, 28, and 42 were collected before reintroducing material from around the pen or adding any new material. Bedding samples were taken with plastic gloves to remove material from 5 subsampling sites in each pen (4 around the perimeter and 1 from the center) using paper cups (Dixie, Fort James, Norwalk, CT). These subsamples from each pen (from 100 to 140 g of the bedding materials) were composited and placed into sterile plastic bags. Upon arrival at the laboratory, 20 g of the bedding sample were removed and analyzed for DM by drying in an oven at 100°C for 24 h. Then 5 g (ambient weight) was removed and mixed with 45 mL of deionized water for 20 min to analyze pH with a pH meter (Denver Instrumental Co., Denver, CO), then left for 3 h to allow absorption of water. After 3 h, excess water was poured from the sample and a wet weight was obtained (Zehner et al., 1986). Absorbency was calculated by the following equation: absorbency = ([wet weight ambient weight] x 100)/ambient weight.
Bacterial populations in bedding were evaluated by weighing 10 g of fresh bedding sample into 90 mL of sterile PBS at pH 7.2 in sterile stomacher bags, then mixing using a Stomacher model 400 mixer (Seward, London, England) at low speed for 1 min. A serial dilution of 1:10 was aspirated with 1 mL of solution into 9 mL of sterile PBS per tube. Each dilution starting at 1 x 103 to 1 x 107 was plated in each of 2 duplicate plates on the surface of the medium. MacConkey agar (Remel, Lenexa, KS) was used to identify gram-negative bacteria and coliform. Bacteria were counted after being incubated for 24 h at 37°C and recorded as cfu/ g of fresh weight. Plates with 30 to 300 colonies were used to calculate cfu/g, and the numbers were converted to logarithms.
Temperature of the bedding surface was measured weekly. Five points of each pen were measured by infrared thermometer (Raynger ST, Santa Cruz, CA) for temperature beginning at 1300 h and before adding any new bedding material. Temperature and relative humidity, measured weekly with 2 wireless thermo-hygrometers (RadioShack, Ft. Worth, TX) averaged 29.9°C and 48.8% inside the barn compared with 32.1°C and 45.7% outside the barn.
The concentration of ammonia from the bedding was quantified by dosimeter gas detection tubes that detected a range of 2.5 to 1000 mg/L (Zefon International, Inc.; St. Petersburg, FL) when calves were 6 wk of age. After the removal of calf, a single gas detection tube was placed in the middle of each pen approximately 10 cm from the top of bedding materials for a 24-h period.
Statistical Analyses
Final body weight, growth, DMI, and feed efficiency data were analyzed as a randomized complete block design by analysis of GLM covariance using initial weight as a covariant (SAS, 1999). Calves were blocked by the week in which they began the study. The DM, absorbency, hardness, and ammonia concentrations data were analyzed using a randomized complete block design. Least-square means were compared with an F-protected (P < 0.05) t-test. Scour days, cleanliness score, fecal fluidity score, blood data, calf behavior, bedding score, pH, bedding temperature, bacterial counts, and bedding material additions were analyzed with PROC MIXED (SAS, 1999). Analyses of these data used a repeated measures model that included effects of subject (calf), bedding material types, time, and the first-order interaction (bedding material types x time). A random statement included block and the block x treatment interaction. Weekly means were calculated and analyzed for BW gain, starter intake, milk replacer intake, scour days, fecal fluidity score, and cleanliness. Significance was declared at P < 0.05.
| RESULTS AND DISCUSSION |
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The average initial and final BW of calves were not different among bedding types (Table 1
). The DMI varied among bedding types during wk 2 due to differences in starter intake. Calves bedded with wood shavings had lower starter intakes than those bedded with granite fines and rice hulls. The average BW gains for d 0 through 42 on all bedding types was less than normal standards (658 g/d; Davis and Drackley, 1998). Fat concentration of milk replacer was only 14.4% by analysis, and may explain the low BW gains. Higher fat in milk replacers has improved BW gain (Scibilia et al., 1987).
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Calf cleanliness scores varied (P < 0.05) among bedding materials (Table 2
). Calves were dirtiest when granite fines were used. Manure was mixed with or absorbed by rice hulls, long wheat straw, wood shavings, and sand, whereas feces tended to remain on the surface of the granite fines because they were packed after use. The rice hulls tended to adhere to the calves more than other materials. Calves skin appeared wetter when bedded with sand and had material near the tail area when bedded with long wheat straw compared with calves bedded on other materials. Calf cleanliness scores varied (P < 0.05) with time (Table 2
). Calves were cleanest during wk 1 and were dirtiest during wk 6. Calf cleanliness scores did not differ during wk 2 to 5.
Physiological Responses
Serum IgG concentration was higher (P < 0.05) at 6 wk of age than earlier but was not affected by bedding materials (Table 3
). The IgG concentration on d 1 of age (958 mg/dL) was less than the recommended 1000 mg/dL (Quigley et al., 2001; Holloway et al., 2002). McGuire and Adams (1982) reported that the adequate level of IgG concentration should be greater than 1600 mg/dL. This failure of passive transfer may explain the increased fecal scores during wk 2 and the lower than expected BW gains for calves in the study (Quigley et al., 1995). Colostral IgG concentration is an important factor in the calfs acquisition of passive immunity, and the rate of absorption of IgG increases with increasing colostral IgG concentration. Therefore, feeding double the recommended rate of colostrum using an esophageal tube may reduce the rate of failure of passive transfer (Cabell, 2002). Immunoglobulin G has to be present in the gut to protect calves from viral enteritis during the first 2 wk of age (Snodgrass et al., 1982; Castrucci et al., 1984; Saif and Smith, 1985).
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There were differences (P < 0.05) by week for percentages of neutrophils and lymphocytes (data not shown). However, bedding treatment did not affect differential white blood cell counts. As calves ages increased, the percentage of neutrophils for all bedding types decreased while the percentage of lymphocytes increased. There were no changes in the percentages of monocytes, eosinophils, and basophils due to age or bedding type.
Serum AGP concentration is a marker of disease and inflammation. Its production is stimulated in hepatocytes by the release of cytokines such as interleukin-1, interleukin-6, and tumor necrosis factor-
from macrophages and monocytes. Higginbotham and Stull (1997) and Itoh et al. (1993) reported that average serum AGP concentrations for neonatal calves were 1172 and 1368 µg/mL, respectively, levels similar to our results. In those studies AGP gradually decreased to 176 ± 28 µg/mL (Itoh et al., 1993) and 815 µg/mL (Higginbotham and Stull, 1997) compared with our data, 613 µg/mL. Our data also showed that AGP was greatest on d 1 and declined with calf age. Bedding treatments had no effect (P > 0.05) on AGP.
Behavioral Data
The percentage of time for self-grooming of calves was affected by bedding materials. Calves housed on rice hulls (Figure 1
) and sand had greater self-grooming behavior than those housed on long wheat straw. Rice hulls adhered to the calves skin, and this may account for the tendency of these calves to conduct more self-grooming. No differences among bedding materials for calf behavioral traits of standing, eating, drinking, investigating environment, and contacting pen were observed.
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On d 42 the concentration (mg/kg per hour; P < 0.05) of ammonia at 10 cm above the bedding surface for a 24-h period was 1.1 for long wheat straw, 1.6 for sand, 1.7 for rice hulls and wood shavings, and 1.9 for granite fines (SE = 0.15). The concentrations of ammonia observed had no effect on calf well being. No respiratory diseases were observed. An ammonia threshold limit value of 15 mg/kg is reported by Seedorf and Hartung (1999). Long wheat straw may have had the lowest ammonia concentration because it had better drainage than other materials (Airaksinen et al., 2001).
Bedding score declined (P < 0.05) from 1.1 on d 0 to 2.5 on d 42. Sand (Table 5
) was the dirtiest, whereas rice hulls, long wheat straw, and wood shavings stayed cleaner because of the particle size of the bedding materials. Inorganic bedding materials (granite fines and sand) had smaller particle sizes than organic bedding materials (rice hulls, long wheat straw, and wood shavings). Small particles would have a greater chance of mixing bedding and the manure than large particles. Sand, rice hulls, and wood shavings mixed with the manure; granite fines appeared to keep the manure on the surface, possibly because granite fines were packed after calves occupied the pen; long wheat straw more readily covered manure after calves occupied the pen. In addition, the absorbency of organic bedding materials (rice hulls, long wheat straw, and wood shavings) was greater than that of inorganic bedding materials (granite fines and sand).
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Surface temperature was affected by bedding materials and time (P < 0.05; Table 5
). As time passed, temperature of all bedding materials declined from 28.5°C on d 0 to 24.4°C on d 42, possibly because the bedding materials absorbed urine; in addition, the air temperatures decreased in the final 3 wk of the study. Long wheat straw was warmest, and rice hulls and wood shavings were warmer than granite fines and sand. The surface temperatures and pH values were usually adequate for gram-negative bacteria and coliform survival (Jay, 2000).
Gram-negative bacteria and coliforms (Table 5
) showed different bedding material by week interactions (P < 0.05). Before use, gram-negative bacteria counts were ranked: rice hulls and wood shavings greater than granite fines, sand, and long wheat straw. Initial coliform counts were ranked: rice hulls greater than wood shavings, sand, long wheat straw, and granite fines. After calves occupied the pen, gram-negative bacteria counts for all beddings and coliform counts for all beddings except rice hulls increased (P < 0.05) on d 14. Gram-negative bacteria counts on d 28 and 42 did not differ from d 14 for any bedding materials. Coliform counts on d 14 for all bedding except rice hulls were higher than those on d 28 and 42. On d 42, long wheat straw had the highest coliform counts. Long wheat straw had greater amino acids, no inhibitory substrates (Bey et al., 2002), lower C:N ratio (Ward et al., 2000), and higher absorbency for bacteria growth compared with wood shavings.
Cost of Bedding Material Types
Weight of materials used per pen (Table 5
) was greatest for granite fines, followed by sand, with rice hulls, long wheat straw, and wood shavings requiring the least material. Using the retail price, the cost of bedding materials per pen from d 0 through 42 were calculated. Long wheat straw ($1.01) and sand ($0.97) were the most expensive. The cost for granite fines ($0.53) was greater than that for wood shavings ($0.24), with rice hulls ($0.07) being the least expensive. However, regional availability and the required transportation distance for each material will significantly affect the on-farm price.
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| ACKNOWLEDGEMENTS |
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Received for publication March 4, 2004. Accepted for publication July 23, 2004.
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