Brief Summary of Minutes of Annual Meeting

Minutes Summary August 12th, 2008 1) Introduction  Terry Engle (host of the meeting) called to order the 2008 Cooperative Western Regional Research Project W-1173 and S-1023 Meeting at 8:30 a.m. a. Attendance was taken and self introductions were made. 2) Administrative reports a. Terry Engle  Chair of the W-1173 group made some general introductory comments about the format of the meeting i. Format  6-15 minute presentations of station reports ii. Poster presentations b. Colin Kaltenbach made a few house keeping announcements i. Send reports to Terry Engle ii. Report of meeting needs to be submitted within 60 days of the end of the meeting c. Deb Hammernik i. Presented CSREES updates and statistics (handouts) 3) Presentations 4) Lunch 5) Presentations 6) Poster session 2:30 p.m.  4:30 p.m. 7) 5:00 p.m. Adjourn Collaborations from W-1173 New York Planned Collaborations A. Work with C.N. Lee of the University of Hawaii and Bob Collier of the University of Arizona on a sweating rate, respiration rate and internal body temperature project. B. Work with C.N. Lee of the University of Hawaii and Ann Parkhurst of the University of Nebraska on statistical analyses of data on the effectiveness of anchors for continuous measurements of vaginal temperature of dairy cows. C. Work with C.N. Lee of the University of Hawaii, Bob Collier of the University of Arizona and Ann Parkhurst of the University of Nebraska on statistical analysis of data to determine if there is cyclic pattern on sweating rate. Mississippi Planned Collaborations Schmidt (MS) is currently in the process of working with Dr. Jeff Carroll (USDA-ARS-TX) to determine the consequences of transportation stress on physiological and nutritional parameters in beef cattle. This project is scheduled to start in mid-October 2008. Schmidt (MS) is working on an evaluation of energy repartitioning of cattle exposed to an immune challenged and transportation stress is also a collaborative research project with Jeff Carroll (USDA-ARS-Lubbock) and Mike Ballou (TX). Willard (MS) is continuing collaborations with Don Lay (USDA-ARS-IN) on the development of biophotonic paradigms for bacterial pathogen tracking in vivo. Willard (MS) is continuing collaborations with Jeff Carroll (USDA-ARS-TX) and Tom Welsh (TX) on the interactions of temperament and stress responses in beef cattle. Hawaii Planned Collaborations AZ, NY and HI plan to study the effect of various solar loads on evaporative cool in controlled environment using the modified portable calorimeter. NB, NY and HI will use statistical model to understand the physiological responses of beef heifers in feedlot conditions. Missouri Planned Collaborations Work with Cheryl Kojima and John Waller of the University of Tennessee, and Jim Strickland and Glen Aiken from USDA/University of Kentucky to identify genetic and physiological markers of sensitivity to heat stress and fescue toxicosis. Work with Bob Collier and Terry Mader on Developing a Program for a Discover Conference on Heat Stress in 2010. Work with Ann Parkhurst on analyzing data for critical breaking points and hysteresis activity during exposure to heat stress. Work with Wayne Kellogg of the University of Arkansas to determine heat stress responses of dairy cows in farm environments in an effort to identify markers of sensitivity. Arizona Planned Collaborations: Work with C.N. Lee of the University of Hawaii, Peter Hillman and Kifle Gebremdehin of Cornell University on a sweating rate project at the University of Arizona Work with Don Spiers and Terry Mader on Developing a Program for a Discover Conference on Heat Stress in 2010. Work with Ann Parkhurst on analyzing data for THI index Kansas Planned Collaborations: Planned collaborations were submitted at the annual meeting. Kansas State plans to collaborate with MARC in understanding how heat stress may alter the porcine intestinal epithelium and affect barrier function in the gut. Tennessee Planned Collaborations TN will collaborate with W-1173 members from USDA-TX and USDA-IN in measuring corticosteroid-binding globulin in plasma from cattle as a physiological measure of stress. TN will collaborate with W-1173 members from USDA-TX, CO and FL on a study examining the effects of weaning strategy on the physiology and performance of beef calves. TN will collaborate with W-1173 members from MO examining genetic markers for fescue toxicity. Indiana Planned Collaborations Work with C.N. Lee of the University of Hawaii on the effect of heat events experienced by the dam on calf health and growth. Work with Don Spiers (MO). on the effect of heat events experienced by the dam in a controlled chamber environment on calf health and growth. Work with John Arthington (FL) on changes in expression of acute phase proteins for cows housed on rubber or concrete. Work with Hank Kattesh (TN) on prenatal stress effects of swine. Colorado Planned Collaborations CO will collaborate with W-1173 members from USDA-TX, TN and FL on a study examining the effects of weaning strategy on the physiology and performance of beef calves.

New York Accomplishment: Sweating rates from heat-stressed dairy and feedlot cows were measured using a Portable Calorimeter and a Bovine Evaporation Meter designed and fabricated for the studies reported herein. Measurements were taken when cows were in their natural habitat. The focus of the study was to compare sweating rates measured from different breeds of dairy and feedlot cows, and determine the level of influence of environmental factors (air temperature, relative humidity, solar load , air velocity), and hair-coat color on sweating rate. The cows were exposed to solar radiation greater than 500 W/m2 (average 833 ± 132 W/m2), average THI was 82.7 ± 1.64 for all studies except for the Nebraska data where the THI was 77.4 ± 4. Air velocity in the sample area was between 0.8 and 1.2 m/s, and body (rectal) temperature was greater than 38.8°C (threshold for heat stress). The range of sweating rates was between 189 ± 84.6 and 522 ± 127.7 g/m2-h, and that of body temperature was between 39.3 ± 0.53 and 41.7 ± 0.19 °C. Differences in sweating rates were statistically significant at P <0.05 value between breeds, between black and white hair coats (within breed) and changes in solar load, relative humidity, and air velocity. Wetting the skin surface coupled with increased air velocity profoundly increased the rate of evaporation by converting sensible heat to latent heat. Behavioral activities of 32 heifers in pens with shade and no shade (exposed to sunlight) were observed to establish whether or not providing shade made a difference in vaginal temperatures of heifers. Four breeds of heifers, 8 of each breed: black Angus, white Charolais, tan-colored MARC I and dark-red colored MARC III were used to determine the effect of hair-coat color on thermal responses. A mechanistic coupled heat and mass transfer model that uses, as an input, environmental parameters and physical and optical properties of hair and hair coat was used to do thermal analysis. There was no significant difference in thermal responses between breeds when heifers were in shade or no shade at an ambient temperature of 36°C. Hair color played a significant role in solar radiation absorption. The black and dark-red colored heifers absorbed higher solar radiation than white and tan colored heifers. Providing shade made a difference in thermal response when the heifers were exposed to 42°C but not when exposed to 36°C. Sensible heat load was sensitive to changes in ambient air temperature, air velocity and density of hair coat. Mississippi Accomplishment (Schmidt): Crossbred calves (n=133) weighing an average of 228.82 ± 22.15 kg were evaluated upon arrival at a 10,000-head capacity backgrounding facility to evaluate an easily obtainable measurement taken at arrival as a prognostic tool for sorting incoming cattle. Standard protocol upon arrival was individual ID (ear tag), antiemetic application and vaccinations for respiratory disease (modified live vaccine) and clostridial pathogens. In addition to arrival protocol, heifers were weighed, rectal temperature determined, and a thermal image (FLIR ThermaCAM EX320) was captured. Thermal images were collected at a standard distance of 76.2 cm from the restraining chute. To determine the correlation between rectal temperature and thermal images; the image of Planum Nasolabiale Area (PNA) of the muzzle was highlighted and then bisected by one vertical line, one horizontal line, and two diagonal lines into quadrants that intersected at the geometric center of the PNA. Muzzle temperature was positively correlated with rectal temperature taken at the time of processing (P<0.01). The most highly correlated area on the muzzle was the convergence of lines in the geometric center of the PNA (r =0.50; P<0.01) and the least correlated measure was the diagonal line bisecting the PNA (r=0.34; P<0.01). Factor one from principle component analysis (PCA) had high loadings for quadrants bisected by the vertical line (0.99 and 0.97; PCA); while maximum temperature of the NPA. Accomplishment (Willard-MS and Lay-USDA-ARS-IN): Acquiring a highly stable photonic plasmid in transformed Salmonella typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella typhimurium (S. typh-lux) using three different plasmids and characterize their respective photonic properties. In presence of ampicillin (AMP), S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 plasmids exhibited 100% photon-emitting colonies over a 10-d study period. Photon emitters of S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 without AMP selection decreased over time (P < 0.05), representing only 11 ± 1%, 35 ± 1% and 43 ± 1%, respectively, of original photon emitting properties of the bacterial population by d 10. Photonic emissions were positively correlated with bacterial concentration (P < 0.05) for pAK1-lux, pCGLS-1 and pXEN-1 (r = 0.96, 0.98 and 0.82, respectively). When stratified by high, medium and low density bacteria concentrations, photonic emissions for high density populations containing pAK1-lux, pCGLS-1 and pXEN-1 resulted in differences of photonic emissions across a range of bacterial concentrations (1×107 to 1×109 CFU, P < 0.05) with positive correlations (P < 0.05) of (r = 0.72, 0.46 and 0.72, respectively). The correlation of photonic emissions with bacterial concentrations for samples with medium and low density bacteria (pAK1-lux, pCGLS-1, and pXEN-1 plasmids) imaged in tubes were also positively correlated (medium; r=0.69, 0.49, 0.46, low; r=0.90, 0.71, 0.68, respectively; P > 0.05); although photonic emissions across a range of bacterial concentrations were not different (1×104 to 1×106 CFU, P > 0.05). For very low density bacterial concentrations imaged in 96 well plates photonic emissions were positively correlated with bacterial concentration (P < 0.05) for pAK1-lux, pCGLS-1, and pXEN-1 plasmids (r=0.99, 0.99, and 0.96, respectively), and photonic emissions across a range of bacterial concentrations (1×103 to 1×105 CFU) low to high were different in the 96-well plate format (P < 0.05). These data characterize photon stability properties for S. typh-lux transformed with three different photon generating plasmids that may facilitate real-time Salmonella tracking using in vivo or in situ biophotonic paradigms. Accomplishment (Willard-MS): In vitro antimicrobial susceptibility testing requires 24 h to several days for results using various methods. In this study, bioluminescence was used to evaluate antimicrobial susceptibility over time of Salmonella typhimurium pAK1-lux (S. typh-lux). Compounds (gentamicin, tetracycline, methionine and Alimet®) were added to S. typh-lux-containing tubes while other tubes had no compounds added (NA). Aliquots were drawn at 0, 2, 4 and 6 h post-treatment, transferred to 96-well plates; (n=8 wells/treatment) and imaged. Wells were serially diluted, plated on agar and incubated. Agar plates were counted, bacterial concentrations calculated, and images analyzed. Photonic emissions were positively correlated with bacterial concentration (R=0.79, 0.81, 0.90, 0.81, 0.85, 0.79, 0.88 (P<0.05) and 0.62 (P=0.053) for NA1, NA2, NA3, NA4, methionine, Alimet®, gentamicin and tetracycline, respectively. Photonic emissions relative to bacterial population increased (P<0.05) and remained stable from 0 to 6 h for the NA, Alimet® and methionine-treated group and decreased (P<0.05) for gentamicin and tetracycline treated group. In conclusion, bacterial concentrations were highly correlated with photonic emissions, and thus S. typh-lux-based systems may be utilized to rapidly evaluate antimicrobials for susceptibility testing in vitro. Accomplishment (Willard): Intrauterine infections in the bovine can have detrimental effects on reproductive performance, and Escherichia coli (E. coli) is a primary causative pathogen. Our objectives were to characterize the photonic properties of E. coli-Xen14 (a stably transformed E. coli containing the lux operon), and conduct photonic imaging of E. coli-Xen14 from within the excised bovine uterus. E. coli-Xen14 (Xenogen) was grown for 24 h in LB broth with or without Kanamycin (KAN; 30¼g/mL) and placed in an incubated shaker at 37°C. Inoculums were placed in 96-well plates for imaging, and photonic emissions collected for 2 s. Following imaging, inoculums were sub-cultured over 7 d, and plated daily (on LB agar plates with or without KAN) to determine the CFU/ml and ratio of photonic emitters vs. non-emitters. Excised bovine reproductive tracts (n = 4 tracts; 3 replicates per tract) were acquired and E. coli-Xen14 placed in a 1 mL tube for photonic imaging (5 s) pre- and post-insertion into the uterine horns. The CFU/ml did not differ (P>0.05) over time with or without KAN presence; remaining stable with 99.93 and 99.98% photonic emitters, respectively. However, RLU/s were lower (P<0.0001) in the presence of KAN than Controls (629.8±117.7 vs. 3,012.0±423.5 RLU/s, respectively). Photonic emissions from E. coli-Xen14 imaged within the uterine horns resulted in a 4,847.1±215.3 RLU/s loss (P<0.05) through the uterine horns, or an efficiency of photonic detection of 3.98±0.007% of the original (pre-) emission intensity. The concentration of photonic E. coli-Xen14 detected through the uterine horns was 662.5 x 106±149.0 x 106 CFU/ml, with a uterine horn wall thickness of 6.2±4.2 mm. In summary, E. coli-Xen14 remained stable with respect to the percentage of photonic emitters with or without KAN (used to selectively culture E. coli-Xen14), however KAN presence suppressed photonic activity. Imaging of E. coli-Xen14 was feasible through the bovine uterine horns ex vivo, suggesting that further development of this model may facilitate the monitoring of pathogen presence within the reproductive tract of the bovine in vivo. Accomplishment (Willard): Collecting blood samples from livestock poses unique challenges when trying to measure stress hormones. Remote blood collection would permit lessened human interaction and sampling within diverse management environments. The IceSampler" blood collection system (ICE) is designed to promote a hands-off approach to blood collection. Our objective was to test the effectiveness of ICE for the collection of blood samples from unrestrained cattle. Successive (n = 9), 16-sample ICE sessions were initiated using 2 Holstein cows with the ICE system to examine uniformity of blood tube volume and timing precision of sample collection. Cows were catheterized and outfitted with the ICE system. The ICE system was then programmed to sample at 15 min to 1 hr intervals. A stress response was artificially induced post-ICE initiation (ACTH challenge; 0.125 IU/kg BW), and cows allowed to roam freely until sampling was completed. When the sequence was completed, the ICE was replenished with new blood tubes and re-launched. The Holstein cows (n=2) fitted with samplers were used for all sampling sessions. Analysis of blood tube volume, sampling time precision, sampler mechanics (e.g. failed tubes) and cortisol (CORT) were used for statistical analysis. Intra-sampling coefficient of variation (CV%) for precision of sampling time was 0.13%; with only 1.8% of recorded sampling events indicating a deviation from expected sampling time (±5 sec in both cases). Intra-sampling CV% for tube blood volume within sampling run was 25.17%, however across sampling runs the inter-sampling CV% for tube blood volume collected was 83.47%; indicating a high degree of variation in sample volume collection (possibly due to catheter rather than ICE performance). Of 9 sampling sessions, 7 were successful (77.8%). Of the n=112 expected blood samples from the seven completed sessions, n=102 blood samples were collected (91.1%). Elevated CORT post-ACTH was evident in ICE samples, and was greater (P < 0.05) than baseline CORT at 3.5 h post-ACTH (0.22 ± 0.03 ug/dl; Range: 0.001 to 0.69 ug/dl). Automated blood sampling in ambulatory, unrestrained cows with ICE was effective, however efforts to improve uniformity of blood volume collection and greater successful sampler completion are needed. Hawaii Accomplishment: The research stations at NY (Dr. Hillman and Dr. Gebremedhin), FL (Dr. Arthington) and HI conducted joint research to evaluate the adaptation of 3 types of Angus in hot-humid summers of Ona, Florida. Six animals were studied for 3 days; black Angus whose origin was from Kansas (2), black Angus whose origin was from Brooksville, Florida (2) and white Angus from Ona, Florida. The objective was to determine the adaptability of the same breed but of different origin and hair coat color to heat stress. The efficacy of sweating (thus evaporative cooling) was measured with a modified portable calorimeter (developed by Peter Hillman, NY). This instrument has variable air flow which would enable one to determine the relationship of air movement directly over the body surface and sweating rates. The study was conducted in field conditions (pasture) and animals at late term pregnancy were restrained using a portable chute. Solar load was recorded >700W/m2 and the average THI was 82.3 (danger zone). The relative humidity was above 62%. The Florida white had lower dorsal skin temperatures (39.60C), lower rectal temperatures (40.0C)and lower respiration rates (RR, 98.4 breaths/min.). This was followed by the Brooksville black (45.50C, 40.80C, 105.0 RR) and the Kansas black (45.00C, 41.70C and 126.9 RR). Florida whites had more short hair and the hair (both long and short hair) were shorter than the blacks. Sweating rates were lowest for when lowest for the Kansas black and highest for the Florida whites. Evaporative cooling increased with air movement at the skin surface for both Florida whites and Brooksville black but not for the Kansas black. Average sweating rates (g/m2-h) were 379.4, 236.1 and 207.3 for Florida whites, Brooksville and Kansas black respectively. Missouri Accomplishment: A study is being conducted to determine both short- and long-term responses to fescue toxicosis using controlled environmental chamber and pasture exposures, respectively. The experiment consists of Angus steers split into 4 groups. The first group is fed uninfected fescue (E-) in the chambers followed by placement on an E- pasture. The second group is fed endophyte-infected fescue (E+) for the whole experiment. The remaining groups are switched between E+ and E- pastures. Collection of data is complete and is currently being analyzed. We hope to determine if heat stress through the summer will result in adaptation to heat stress (e.g., lower core temperature, respiration rate, and sweat rate) with the E- group. The second group determines if E+ pasture will improve response to E+ under controlled conditions or whether the combination is overwhelming and reduces animal health. Finally the two switch groups allow for testing of whether consuming E+ changes the response to heat stress and if summer heat alters the animals response to an E+ challenge. One preliminary result indicates that sweating rate is severely reduced as a result of fescue toxicosis. Preliminary results from the previous experiment showed that sweating rates were lower as a result of fescue toxicosis. One possible reason for a low sweating rate is animal dehydration. An experiment was performed to determine the effects of dehydration on cattle. Angus steers were acclimated for seven days at thermoneutrality (21C). This was followed by three days each of water restriction and rehydration. Rectal temperature and respiration rate were measured six times daily. Blood samples were taken prior to dehydration, and once daily during dehydration and rehydration. Body weight, food and water intakes, and cutaneous water loss at shaved sites were recorded during acclimation and daily during dehydration and rehydration. Steers lost 4 percent body weight during dehydration, but it returned to baseline within 24 hours rehydration. Feed intake decreased (70 percent) within 24 hours of dehydration, and recovered after 24 hours rehydration. Respiration rate and cutaneous water loss decreased with dehydration, which, combined with the reduction in feed intake, resulted no physiological change in rectal temperature. Hematocrit was a variable indicator of dehydration, decreasing during the first 24 hours, before returning to baseline. Serum osmolarity increased within 24 hours without water, and remained elevated until rehydration. Dehydration appeared more complex than rehydration, resulting in increases in some blood parameters and no change in others. In contrast, rehydration decreased in all blood parameters to suggest blood dilution. The above experiment was done under thermoneutral conditions to concentrate on only one stressor, dehydration. Thermal status was maintained at thermoneutrality which was expected in the absence of thermal stress. A second experiment was performed to determine the effects of heat stress along with dehydration. Angus steers were acclimated for seven days at thermoneutrality before being exposed to 12 days of cyclic daily heat stress (26°C to 36°C). Three days water restriction began on the fourth day of heat stress and was followed by rehydration. Data has been collected, and is currently being analyzed. Preliminary results show large decreases in feed intake and body weight. Surprisingly, sweating rates only showed marginal decreases with dehydration, and core body temperature was maintained for the entire water restriction period. Rats were monitored at thermoneutral and heat stress environments to determine the short-term time-related changes associated with intake of diets containing endophyte-infected (E+) and uninfected (E-) seed. In addition, some rats were pair-fed to the E+ group (PFE+), using the E- seed diet, to determine the effect of reduced caloric intake associated with intake of the E+ diet. This allowed for a separation of the direct effects of fescue toxicosis from indirect effects of fescue toxicosis. The E+ and PFE+ rats reduced their feed intake and daily gains. However, the PFE+ rats had lower core body temperature compared to E+ rats during heat stress. Also, serum prolactin levels were higher in PFE+ than E+ rats in both thermoneutral and heat stress environments. The E+ rats reduced their heat tolerance, whereas the pair-fed rats had improved tolerance. The circadian rhythm of E+ rats under heat stress was altered with a higher core temperature during the inactive period which explains the failure of thermoregulation in this group. Therefore, the reduced feed intake component of fescue toxicosis does not contribute to the hyperthermia associated with fescue toxciosis. Previous studies using rats in our lab have indicated that the reduction in feed intake associated with consumption of endophyte-infected fescue (E+) seed is not due to ergovaline (EV). Ergovaline is one of many ergot alkaloids found in E+ seed. In order to better define the role of EV in the feed intake reduction response, EV alone was rotavapped onto commercial ground rat chow at different concentrations and fed to young adult male rats. The maximum concentration of EV that we have been able to achieve in a diet made with seed has been ~2.06ppm. Pure EV was rotavapped onto a commercial diet at three different levels: 2.06ppm, 3.09ppm, and 4.12ppm. Rats did not display any significant decrease in feed intake at any concentration of EV. They were allowed to recover on plain ground chow, followed by endophyte-free seed diet, then fed a diet containing E+ seed. As we have repeatedly seen, feed intake dropped immediately upon introduction of the E+ seed diet to confirm that these animals were responsive to treatment with ergopeptine alkaloids. The body temperature response to EV rotavapped onto chow will be investigated in future studies. Arizona Accomplishment: The Temperature Humidity Index originally developed by Thom (1958) and extended to cattle by Berry and colleagues (1964) is used to estimate cooling requirements of dairy cattle. However, correlation of THI to measurements of core body temperature is not adequate under moderate climatic conditions because of the large heat increment needed to raise core body temperature. Our objective was to determine if incorporation of skin surface temperature to mean body temperature measures would improve Pearson Correlation Cooefficients between THI and physiological measures of heat stress. During a three-month span, three groups of cows (n=12) were housed in the environmental chambers at the University of Arizona William J. Parker research complex for a total of 22 days. Each group experienced a total of three environments where THI was at a minimum of 49, maximum of 79 and an average 62.9. We utilized NRC 1971 THI calculation (0.18 x Tdb+32)  (0.55-(0.0055 x RH) x (1.8 x Tdb-26) for estimate of environmental heat load. Physiological measures of heat strain included respiration rate (RR/min), infrared surface temperature (ST, ºC), rectal temperature (RT. ºC), heart rate (HR/min), and evaporative heat loss (EVHL, g/m2). Mean body temperature (BT, ºC) was calculated using the formula BT = (0.33 x ST+0 .67 x RT). Respiration rates, ST, RT, EVHL, and HR were routinely collected 4 times per day and hourly for 24 hours once biweekly for a total of 2096 measures. Correlations of these measures to THI were as follows: THI and RR (r=0.40; P<0.0001), THI and RT (R2=0.17; P<0.0001), THI and SR (R2=0.40; P<0.0001), THI and ST (R2=0.74; P<0.0001), THI and BT (R2=0.72; P<0.0001), and THI and HR (R2=0.002; P<0.05). Correlations between BT and RR (R2=0.48; P<0.0001), BT and SR (R2=0.46; P<0.0001), BT and RT (R2=0.39; P<0.0001). We conclude that under moderate climatic THI conditions investigated addition of ST to BT improved the correlation of BT to THI over RT alone. In addition, the correlation of RR, ST, and BT are improved over RT alone implying that ST has a greater effect on RR than RT. Accomplishment: 1. Niacin has been shown to increases resistance to thermal stress in cattle by increasing whole body evaporative heat loss in vivo and cellular heat shock response to thermal stress in vitro. To determine effect of feeding protected niacin (NIASHURE") on body temperature, milk yield and composition a total of 400 lactating primiparous and multiparous Holstein cows were randomly assigned to a switchback design of either control (C, no feed additive n=200) or treatment (Trt, cows supplemented with 12g/d/cow of encapsulated niacin, n=200). Groups were balanced for DIM, milk yields, and parity prior to start of the study which was conducted from August 7th thru October 7th, 2007 on a commercial dairy in Arizona. Cows remained in their respective treatment for 30 days and then switched to the opposite treatment on day 31 and continued until day 60. Milk yields were recorded three times daily and a monthly milk sample was collected to obtain milk components. Feed samples were collected weekly from each group. Vaginal temperatures were collected using iButton (MAXIM) temperature loggers attached to a blank continuous intravaginal drug release device and inserted into a random sub-sample of cows (n=16) from each pen (n=2) with similar DIM, milk yields, and parity for 7 d. Body core temperatures were decreased for the Trt group during periods of peak thermal load from 1300 to 1600 h. Milk fat and protein percent was elevated in the Trt versus C groups (3.65 vs. 3.38 and 3.09 vs. 3.05 %, respectively; P<0.01). Subsequently both fat- and energy-corrected milk was greater for cows in the Trt group compared with cows in the C group (39.7 vs. 38.2 and 39.6 vs. 38.4 kgs, respectively; P<0.01) In addition, both fat- and energy-corrected milk was increased in multiparous compared to primiparous cows (40.7 vs. 37.2 and 40.7 vs. 37.3 kgs; P<0.01). However, groups did not differ in milk yield for either test day (39 vs. 38.8 kg) or average daily milk samples (37.6 vs. 37.5 kg), and group fed DMI (59.4 vs. 59.7 lbs). In conclusion, supplementation of lactating cows with encapsulated niacin during summer heat stress reduced core body temperature and increased both fat and protein percent, in turn, elevating fat- and energy-corrected milk yields. 2. Niacin induces peripheral vasodilatation via prostaglandin D (PGD) and E (PGE) release by Langerhans cells in skin. We evaluated if niacin alone or in combination with PGD and PGE alters expression of heat shock proteins (HSP) 27 and 70. Bovine mammary epithelial cells (BMEC) were cast in collagen in 24-well plates containing growth media (GM) composed of DMEM/F-12, insulin, EGF, IGF-I, BSA and antibiotics at 37°C, 5% CO2. Cultures grew into ductal structures with media changes at 48 hr intervals. On day 8 cultures were divided into Controls (C) receiving GM, GM with niacin (0.5, 1.0, or 10.0 mM), PGD2 (10 or 24 uM), PGD2 with PGE1 (both at 24uM) alone or in combination with niacin. Half were placed into incubators at 37°C (TN) and the remainder at 42°C (HS) for 8 h. At 0h and 8h, replicates were pooled, placed in TRIzol and stored at -80°C until extracted for RNA. Isolated RNA was reverse transcribed into cDNA. Expression of HSP's-27 and 70 was measured by q- PCR. Addition of PGD increased HSP-27 and 70 gene expression in HS, (P<.0001). Peak fold increases in HSP-70 expression at 8 hr over time zero differed between C and PGD, (-2.4 vs. 9.3, P<.0001) and were greater for Hsp-27 (-115 vs +10, P<.0001). Addition of PGE increased HSP-27 and 70 expression compared to C and PGD alone ( P<.05). We conclude that niacin with PGD or PGD+PGE alters HSP-27 and Hsp-70 gene expression in BMEC during HS. Kansas Accomplishment: We have recently focused on the interaction of intestinal epithelial cells with both commensal and pathogenic bacteria. These goals are placed in the broader context of understanding how in-feed antimicrobials modulate intestinal responses that may contribute to the growth responses in weaned pigs commonly seen with antibiotic feeding. To that end, we have been evaluating prototypical probiotic bacteria in a model porcine gut epithelium. One such probiotic bacterium is Bacillus licheniformis (BL). We chose to evaluate BL because it is one of two Bacillus organisms in the BioPlus 2B product. We have now has evidence that BL interacts with swine gastrointestinal epithelial cells to intervene in the inflammatory response to Salmonella enterica serovar Typhimurium (ST; Skjolaas et al., 2007). In the current studies, IPEC-J2 cells were grown on permeable membrane inserts and used in experiments employing a 2 x 2 factorial arrangement of treatments. Cells were pre-exposed apically to 108 CFU BL (or sterile media) followed by apical treatment with 108 CFU ST (or sterile media). The resulting treatments were: 1) control (no bacteria); 2) BL only; 3) ST only; or 4) BL followed by ST. In Exp. 1, cells were pre-exposed to BL 4 h prior to ST treatment, and in Exp. 2 for 8 h. Cells were washed 1 h after ST, and media containing 50 ¼g/mL gentamicin was added to kill remaining extracellular bacteria. Apical and basolateral media were collected 5 h later and assayed for IL8. At the conclusion of the each experiment, IPEC-J2 cells were extracted to estimate percent invasion by ST. In Exp. 1, ST stimulated IL8 secretion compared to control wells (P < 0.001) and pre-exposure with BL blunted ST-stimulated IL8 both apically (P < 0.01) and basolaterally (P < 0.05). Similarly, in Exp. 2, ST stimulated IL8 secretion, and pre-exposure with BL reduced ST-stimulated IL8 secretion apically (P < 0.05) and basolaterally (P < 0.001). Pre-exposure to BL did not reduce ST invasion of IPEC-J2 cells in either experiment. The current studies suggest that short pre-exposure of swine intestinal epithelial cells with BL blunts ST-induced IL8. Inhibition of IL8 by BL appears to increase with increased duration of pre-exposure. The mechanism(s) by which BL reduces IL8 response to ST is not completely understood, but does not appear to reflect an ability of BL to reduce ST invasion of swine epithelial cells. Tennessee Accomplishments: Parameters measuring growth, stress and immune response in weanling pigs given a synthetic opiate agonist (OA) were investigated in three separate trials. Pigs receiving a single intramuscular injection of OA at 13 d of age and weaned 6 d later exhibited increased feed intake (FI) and body weight (BW) compared with controls 3-7 d post-weaning. However, FI and BW were not different between treatments during the 7 d post-weaning when OA was administered on the day of weaning. Plasma cortisol, free cortisol index, and growth hormone increased within 3 h following OA. The number of neutrophils, lymphocytes and monocytes were greater in OA pigs measured on d 2-4 following a bacterial challenge compared with similarly challenged controls. Impact: Administration of a single injection of OA may have beneficial effects on the health and well-being of weanling pigs, but the timing of injection relative to weaning may modulate the response. Indiana Accomplishment: Dairy lameness persists as an economic and well-being concern of dairy producers. To determine the effects of rubber flooring on some indicators of health, blood samples were collected over the first and second lactation from cows housed on free-stall quadrants containing rubber at the feed bunk interface or grooved concrete. The analysis showed that housing the cows on rubber mats altered two genes of interest (MMP13 and TAC1) and leukocyte populations (IN). However, acute phase proteins were not different between weeks 8 and 26 after the second lactation (FL). Accomplishment: Sows housed in gestation stalls develop various abnormal behaviors referred to as stereotypic. Stereotypic behaviors are largely accepted as an indicator of exposure to chronic stress. Exposure to chronic stress has been shown to result in a dysregulation of the hypothalamic-pituitary-adrenal (HPA) response to stress in several species. To determine if sows had an altered function of their HPA axis, we subjected stall housed sows to a dexamethasone suppression test. This test has been widely used to diagnose depression and anxiety in humans. The test is conducted by taking a baseline blood sample from a patient in the late afternoon and then administering a low dose of dexamethasone. A second blood sample is collected 12 hours later to measure glucocorticoids. The DST did not prove to separate the physiologic response of sows to dexamethasone based on the performance of abnormal oral behavior. However, 4 of the 23 sows were resistant to glucocorticoid suppression by dexamethasone. Colorado Accomplishments: Two hundred and sixteen Angus crossbred steers purchased from sale barns (230 kg ± 3.6) were utilized to determine the impact of trace mineral source and concentration on performance, tissue metabolites, and lipid metabolism. Treatments during the 27 d receiving phase consisted of: 1) Inorganic trace mineral (INORG; 125 mg CuSO4/d per head; 360 mg ZnSO4/d per head; 200 mg MnSO4/d per head; and 12.5 mg CoCO3/d per head) and 2) Organic trace mineral (ORG; iso-amounts of Cu, Zn, Mn amino acid complexes, and Co glucoheptonate). On d 0 and 27, blood samples were collected from three steers per pen (pen = experimental unit). On d 28, steers were transitioned to a high concentrate finishing diet containing different concentrations of Co. Treatments during the finishing phase consisted of: 1) control (no supplemental Co); 2) 0.10 mg Co/kg DM from cobalt glucoheptonate; 3) 1.0 mg Co/kg DM from cobalt glucoheptonate. The same three steers per pen were bled on d 27, 84, and 224 of the finishing phase. During the receiving phase, red blood cell superoxide dismutase activity was greater (P < 0.03) for ORG vs. INORG supplemented steers. During the finishing phase, overall ADG tended (P < 0.06) to be greater for steers receiving 1.0 mg Co/kg DM. Steers receiving 1.0 mg Co/kg DM had greater YG (P < 0.04) and back fat thickness (P < 0.04) than steers receiving 0.10 mg Co/kg DM. Serum, liver, and longissimus muscle B12 concentrations increased (P < 0.04) as dietary Co concentration increased. The objective of this study was to characterize pre-stun handling of commercial swine (n=76) and explore its relation to exsanguination blood lactate concentration ([lactate]) and rectal temperature in a commercial processing plant. One observer scored pig behavior and handling in the crowd pen and single file chute leading to the V-restrainer. Handling scores were tabulated as the number of times each pig experienced one of the target handling events. Exsanguination [lactate] (mM) and rectal temperature (F) were measured following electrical stunning. Behavioral data were analyzed using Pearson product moment correlations in SAS 9.1 (SAS Inst., Inc., Cary, NC) to determine the relationship between the behavioral and physiological measures. Results indicated positive correlations between [lactate] and jamming (r=0.23, P=0.05), backing up (r=0.27, P<0.05) and rearing (r=0.25, P<0.05). Rectal temperature was correlated with the duration of time spent in the handling area (r=-0.26, P<0.05). Single degree of freedom contrasts were used to compare differences in exsanguination [lactate] and rectal temperature based on the presence or absence of handling events. Pigs that experienced electric prod use, jamming, rearing and/or backing up had a greater (P=0.05) [lactate] (7.8 ± 0.43) than animals that did not experience these events (6.2 ± 0.69). Pigs that experienced jamming, rearing and/or backing up had a greater (P<0.05) [lactate] (8.3 ± 0.48) than animals that did not (6.2 ± 0.52). Pigs that jammed had a greater (P<0.05) [lactate] (8.5 ± 0.68) than animals that did not (6.9 ± 0.43). Pigs that reared and/or jammed and animals that only reared exhibited a trend towards greater (P=0.11 and P=0.06, respectively) [lactate] than pigs that did not experience those events. No differences in rectal temperature were demonstrated between the aforementioned contrast groups. Aspects of pre-stun animal handling can help to explain exsanguination [lactate].

Mississippi: Ryan P., K. Moulton, S. Willard, J. Crouch, D. Sykes, D. Christiansen and W. Bennett. 2008. Experimentally-induced placentitis in late gestation ewes and mares: evaluation of pathogen progression using lux-modified bacteria and bioluminescence imaging. In: Uterine infections in Mares and Women: A comparative Study II, eds M. LeBlanc, J.F. Wade and L. Foster, Havemeyer Foundation Monograph Series #19, R and W Communications, Newmarket, UK, pp 35-36. Moulton, K., P. Ryan, D. Christiansen, R. Hopper, C. Klauser, W. Bennett, S. Rodts-Palenik and S. Willard. 2008. Ex vivo bioluminescence imaging of late gestation ewes following intrauterine inoculation with lux-modified Escherichia coli. Comp Immunol Microbiol Infect Dis (Published online ahead of print: doi:10.1016/j.cimid.2008.02.001); published on-line: April 25, 2008. Hawaii: Gebremedhin, K.G., P.E. Hillman, C.N. Lee, R.J. Collier, S.T Willard, J. Arthington, and T.M. Brown-Brandl. 2008. Sweating rates of dairy and feedlot cows in stressful thermal environments. ASABE Paper No. 084752, St. Joseph, MI Presented at the ASBE Annual International Meeting, Providence, RI, June 29  July 2, pp. 29. Gebremedhin, K.G., P.E. Hillman, C.N. Lee, R.J. Collier, S.T Willard, J. Arthington, and T.M. Brown-Brandl. 2008. Sweating rates of dairy and feedlot cows in stressful thermal environments. Proceedings of the Eighth International Livestock Environment Symposium, September 1-5, 2008, Iguassu, Brazil, pp.10. Missouri: Bhusari S., L. B. Hearne, D. E. Spiers, W. R. Lamberson and E. Antoniou. 2007. Expression profiling of heat stress effects on mice fed ergot alkaloids. Toxicological Sciences. 95 (1): 89. Al-Tamimi, H.J., P.A. Eichen, G.E. Rottinghaus, and D.E. Spiers. 2007. Nitric oxide supplementation alleviates hyperthermia induced by intake of ergopeptine alkaloids during chronic heat stress. J. Thermal. Biol. 32: 179. Bhusari, S., L. B. Hearne, D. E. Spiers, W. R. Lamberson, and E. Antoniou. 2008. Transcriptional Profiling of Mouse Liver in Response to Chronic Heat Stress. J. Thermal Biol. 33: 157. Settivari, R.S., T.J. Evans, P.A. Eichen, G.E. Rottinghaus, and D..E. Spiers. 2008. Short- and long-term responses to fescue toxicosis at thermoneutrality or during heat stress. J. Thermal Biol. 33: 213. Settivari, R.S., T. J. Evans, E. Rucker, G.E. Rottinghaus, and D.E. Spiers. 2008. Effect of ergot alkaloids associated with fescue toxicosis on hepatic cytochrome P450 and antioxidant proteins. Toxicology and Applied Pharmacology. (Accepted). Kirch, B.H., G.E. Aiken, and D.E. Spiers. 2008. Temperature influences upon vascular dynamics in cattle measured by Doppler ultrasonography. J. Thermal Biol. (Accepted). Scharf, B., L.E. Wax, G.E. Aiken, and D.E. Spiers. 2008. Regional differences in sweat rate response of steers to short-term heat stress. Int. J. Biometerol. (Accepted). Scharf, B., L.E. Wax, J.A. Carroll, D.G. Riley, C.C. Chase, Jr., S.W. Coleman, D.H. Keisler, and D.E. Spiers. 2008. Evaluation of physiological and blood serum differences in heat tolerant (Romosinuano) and heat susceptible (Angus) Bos taurus cattle for determination of markers of sensitivity. Proceedings of the 8th International Livestock Environment Symposium (ASABE ILES VIII) to be held in September 2008 in Iguaçu Falls, Brazil. Scharf, B., L.E. Wax, T.J. Evans, and D.E. Spiers. Impact of dehydration on production and thermoregulation of angus steers at thermoneutrality. Proceedings of the 8th International Livestock Environment Symposium (ASABE ILES VIII) to be held in September 2008 in Iguaçu Falls, Brazil. Eichen, P.A., M.C. Lucy, T.J. Safranski, E.A. Coate, A.M.Williams, and D.E. Spiers. Heat Stress Effects on Sow Reproductive Performance Using Simulated Forced Air and Evaporative Cooling Systems. Proceedings of the 8th International Livestock Environment Symposium (ASABE ILES VIII) to be held in September 2008 in Iguaçu Falls, Brazil. Arizona: Collier, R.J., M.A. Miller , C. L. McLaughlin , H.D. Johnson , and C. A. Baile .2008. Effects of Recombinant Bovine Somatotropin (rbST) and Season on Plasma and Milk Insulin-Like Growth Factors I (IGF-I) and II (IGF-II) in Lactating Dairy Cows. Domest Anim. Endocrinol Collier, R.J., J. L. Collier, R. P. Rhoads and L.H. Baumgard. 2007. Genes involved in the Bovine heat stress response. J. Dairy Sci.91:445-454. Burgos, R., L.J. Odens, R.J. Collier, L. H. Baumgard, and M.J. VanBaale.2007. Case Study: Evaluation of cooling systems in lactating heat-stressed dairy cows in a semi-arid environment. Prof. Anim. Sci. 23:546-555. Kansas: Burkey, T.E., K.A. Skjolaas, S.S. Dritz, and J.E. Minton. 2007. Expression of Toll-like receptors, interleukin 8, macrophage migration inhibitory factor, and osteopontin in tissues from pigs challenged with Salmonella enterica serovar Typhimurium or serovar Choleraesuis. Vet. Immunol. Immunopathol. 115:309-319. Corrigan, M. E., J. S. Drouillard, M. F. Spire, D. A. Mosier, J. E. Minton, J. J. Higgins, E. R. Loe, B. E. Depenbusch, and J. T. Fox. 2007. Effects of melengestrol acetate on the inflammatory response in heifers challenged with Mannheimia haemolytica. J. Anim. Sci. 85:1770-1779. Farran T. B., C. D. Reinhardt, D. A. Blasi, J. E. Minton, T. H. Elsasser, J. J. Higgins and J.S. Drouillard. 2008. Source of dietary lipid may modify the immune response in stressed feeder cattle. J. Anim. Sci. 86:1382-1394. Fraser, J.N., B. L. Davis, K. A. Skjolaas, T. E. Burkey, S. S. Dritz , B. J. Johnson, and J. E. Minton. 2007. Effects of feeding Salmonella enterica serovar Typhimurium or serovar Choleraesuis to weaned pigs on growth performance and circulating insulin-like growth factor-I, tumor necrosis factor alpha, and interleukin-1 beta. J. Anim. Sci. 85:1161-1167. Skjolaas, K.A., T.E. Burkey, S.S. Dritz, and J.E. Minton. 2007. Effects of Salmonella enterica serovar Typhimurium, or serovar Choleraesuis, Lactobacillus reuteri and Bacillus licheniformis on chemokine and cytokine expression in the swine jejunal epithelial cell line, IPEC-J2. Vet. Immunol. Immunopathol. 115:299-308. Williams, J. L., J. E. Minton, J. A. Patterson, J. Marchant Forde and S. D. Eicher. 2008. Lairage during transport of eighteen-kilogram pigs has an impact on innate immunity and commensal bacteria diversity in the intestines. J. Anim. Sci. 86:1232-1244. Woodworth, J. C., M. D. Tokach, J. L. Nelssen, R. D. Goodband, S. S. Dritz, S. I. Koo, J. E. Minton, and K. Q. Owen. 2007. Influence of dietary L-carnitine and chromium picolinate on blood hormones and metabolites of gestating sows fed one meal per day. J. Anim. Sci. 85:2524-2537. Tennessee: Doherty, T. J., H. G. Kattesh, R. J. Adcock, M. G. Welborn, A. M. Saxton, J. L. Morrow, and J. W. Dailey. 2007. Effects of a concentrated lidocaine solution on the acute phase stress response to dehorning in dairy calves. J. Dairy Sci. 90:4232-9. Lay, D. C. Jr., H. G. Kattesh, J. E. Cunnick, M. J. Daniels, K. A. McMunn, M. J. Toscano, and M. P. Roberts. 2008. Prenatal stress on pig development and response to weaning. J. Anim. Sci. 86:1316-24. Kojima, C. J., H. G. Kattesh, M. P. Roberts, and T. Sun. 2008. Physiological and immunological responses to weaning and transport in the young pig: modulation by administration of porcine somatotropin. J. Anim. Sci. doi:10.2527/jas.2008-1089. Indiana: Kanaan, V.T., E.A. Pajor, D.C. Lay Jr., B.T. Richert, and J.P Garner. 2008. A note on the effects of co-mingling piglet litters on pre-weaning growth, injuries and responses to behavioural tests. Appl. Anim. Behav. Sci. 110 (3), p.386-391. Kranendonk, G., D.C. Lay Jr., S. Jarvis, M. Fillerup, C.G. VanReenen, H. Hopster, and V.M. Wiegant. 2008. Regular mixing of pregnant sows reduces their weight gain, but does not affect offspring body weight, behaviour or wound healing. Appl. Anim. Behav. Sci. Accepted with revision. Bearson, B. L., S. M. D. Bearson, J. J. Uthe, S. E. Dowd, J. Houghton, I. Lee, M. J. Toscano, and D. C. Lay Jr. 2008. Iron regulated genes of Salmonella enterica serovar Typhimurium in response to norepinephrine and the requirement of fepDGC for norepinephrine-enhanced growth. Microbes and Infection. 10:807-806 Lay, D. C. Jr., H. G. Kattesh, J. E. Cunnick, M. J. Daniels, K. A. McMunn, M. J. Toscano, and M. P. Roberts. 2008. Effects of prenatal stress on sow productivity and piglet response to weaning. J. Anim. Sci. 86:1316-1324. Schenck, E.L., McMunn, K.A., Rosenstein, D.S., Stroshine, R.L., Nielsen, B.D., Richert, B.T., Marchant-Forde, J.N. and Lay Jr., D.C. 2008. Exercising stall-housed gestating gilts: Effects on lameness, the musculo-skeletal system, production and behavior. J. Anim. Sci. In Press. Marchant-Forde, J.N., D.C. Lay Jr., R. Marchant-Forde, K.A. McMunn, and B.T. Richert. 2008. Effects of r-salbutamol on behavior and physiology of finishing pigs. J. Anim. Sci , In Press. Wilcox, C.S., N.M. Schutz, S.S. Donkin, D.C. Lay Jr., and S.D. Eicher. 2008. Short Communication: Effect of temporary glycosuria on molasses consumption in Holstein calves. J. Dairy Sci. 91:1-4. Williams, J. L., J. E. Minton, J. A. Patterson, J. N. Marchant Forde, and S. D. Eicher. Lairage during transport has an impact on 18 kg pigs innate immunity and commensal bacteria diversity. 2008. J. Anim. Sci. 86:1232-1244. Colorado: Gruber, S. L. J. D. Tatum, T. E. Engle, K. J. Prusa, S. B. Laudert, A. L. Schroeder, and W. J. Platter. 2008. Effects of ractopamine supplementation and postmortem aging on longissimus muscle palatability of beef steers differing in biological type. J. Anim. Sci. 86: 205-210. Legleiter, L. R., J. K. Ahola, T. E. Engle, and J. W. Spears. 2007. Decreased Brain copper due to copper deficiency has no effect on bovine prion proteins. Biochem. Biophy. Res. Comm. 352:884-888. Dorton, K. L., T. E. Engle, R. M. Enns, and J. J. Wagner. 2007. Effects of trace mineral supplementation,source, and growth implants on immune response of growing and finishing feedlot steers. Prof. Anim. Sci. 23:29-35. Whitman, K. J., T. E. Engle, P. E. Burns, K. L. Dorton, J. K. Ahola, R. M. Enns. 2007. Effects of copper and zinc source on performance, carcass characteristics, and lipid metabolism in finishing steers. Prof. Anim. Sci. 23:36-41. Fulwider, W. K., T. Grandin, D. J. Garrick, T. E. Engle, W. D. Lamm, N. L. Dalsted, and B. E. Rollin. 2007. Influence of free-stall base on tarsal joint lesions and hygiene in dairy cows. J. Dairy Sci. 90: 3559-3566. Gruber, S. L., J. D. Tatum, T. E. Engle, M. A. Mitchell, S. B. Laudert, A. L. Schroeder, and W. J. Platter. 2007. Effects of ractopamine supplementation on growth performance and carcass characteristics of feedlot steers differing in biological type. J. Anim. Sci. 85: 1809-1815. Schneider, B. A., J. D. Tatum, T. E. Engle, and T. C. Bryant. 2007. Effects of heifer finishing implants on beef carcass traits and longissimus tenderness. J. Anim. Sci. 85:2019-2030.