Donald E. Spiers

Profile of the bovine acute-phase response following an intravenous bolus-dose lipopolysaccharide challenge.

Our objective was to characterize further the acute-phase response following endotoxin (i.e. lipopolysaccharide; LPS) exposure in the bovine. Nine pure-bred Angus castrated males (i.e. steers; average body weight = 299 ± 5 kg) were used in a randomized complete block design in environmentally controlled chambers, set at thermoneutral level, to characterize the acute physiological, endocrine, immune, and acute-phase protein responses following an i.v. bolus administration of 2.5 μg of LPS/kg body weight. One day before administration of LPS, all steers were fitted with an indwelling jugular vein catheter for serial blood collection. Blood samples were collected at 30-min intervals from -2 h to 8 h relative to the LPS challenge (time 0), and serum was harvested and stored at -80 °C until analyzed for concentrations of cortisol, pro-inflammatory cytokines, and acute-phase proteins. Indicators of thermal status (i.e. rectal temperature, ruminal temperature, respiration rate, sweat rate, and skin temperatures) were measured at 30-min intervals from -1 h to 6 h relative to the challenge. Endotoxin exposure increased (P<0.05) serum concentrations of cortisol, tumor necrosis factor-α (TNF-α), interleukin 1-β (IL-1β), IL-6, interferon-γ (IFN-γ), and serum amyloid A. Respiration rate, rectal temperature, and rump skin temperature also were increased (P<0.05) following LPS administration. Endotoxin exposure dramatically decreased ear skin temperature (P = 0.002), but tended to increase (P<0.10) ruminal temperature, shoulder skin temperature, and shoulder sweat rate. Serum concentrations of acid soluble protein, α-acid glycoprotein, IL-4 and IL-2, and rump sweat rate were not altered (P>0.24) by the challenge. To our knowledge, this report is the most complete characterization of the bovine acute-phase response to a bolus-dose endotoxin challenge conducted under thermoneutral conditions and should provide foundation data for future research.

Evaluation of physiological and blood serum differences in heat-tolerant (Romosinuano) and heat-susceptible (Angus) Bos taurus cattle during controlled heat challenge.

A study was performed to evaluate differences in thermoregulatory ability of 2 Bos taurus breeds with known differences in heat tolerance. Nine Angus (AG; 304 +/- 7 kg of BW) and 9 Romosinuano (RO; 285 +/- 7.5 kg of BW) steers were transported to the Brody Environmental Center at the University of Missouri. Steers were housed for 18 d at thermoneutrality (TN; 21 degrees C) before initiation of heat stress (HS), which consisted of daily cyclic air temperature (26 degrees C, night; 36 degrees C, day) for 14 d. Rectal temperature and respiration rate were measured 6 times daily throughout the study. Sweat rates at shaved skin sites were recorded on specific days. Blood samples were taken once per week. Angus steers maintained rectal temperature 0.5 degrees C greater than RO at TN (P < 0.001). Likewise, respiration and sweat rates were greater (P < 0.001) in AG than RO at TN (P < 0.05). Rectal temperature increased during HS for both breeds with AG maintaining greater temperatures (P < 0.001). Both breeds increased respiration rate during HS, with AG steers exhibiting the greater rate (P < 0.001). Sweat rate increased more than 4-fold during HS (P < 0.001), followed by reduction after 7 d. Even after HS acclimation, AG exhibited the greater sweat rate (P < 0.001). Breed differences for serum leptin, creatinine, and cholesterol were found throughout the study with AG being greater than RO. Although there were no breed differences (P = 0.21) at TN, only AG steers exhibited a HS-induced increase (P < 0.05) in prolactin, creatinine, and cholesterol concentrations to suggest that an increase in rectal temperature is required for this effect. Use of rectal temperature along with endocrine markers, such as prolactin, may aid in the identification of B. taurus sensitivity to heat.

Effects of a controlled heat stress during late gestation, lactation, and after weaning on thermoregulation, metabolism, and reproduction of primiparous sows

Heat stress (HS) causes seasonal infertility in sows and decreases reproductive efficiency. The objective was to examine thermoregulation, metabolic responses, and reproduction in sows exposed to HS or thermoneutral (TN) conditions during different phases of a production cycle (gestation, lactation, and breeding). Fifty-eight first-parity Landrace (n = 26) or Landrace × Large White F1 (n = 32) sows were rotated through environmental chambers for 57 d beginning in late gestation. The ambient temperature sequences included either TN (18°C to 20°C) or HS (24°C to 30°C) for each production phase with the following treatment groups: TN-TN-TN (n = 15), TN-HS-TN (n = 14), HS-TN-HS (n = 14), and HS-HS-HS (n = 15) for gestation-farrowing-breeding (20, 24, and 13 d, respectively). Regardless of the temperature treatment, rectal temperatures were greater (P < 0.001) during lactation (39.36°C ± 0.01°C) than during the gestation (38.27°C ± 0.01°C) or the breeding period (38.77°C ± 0.01°C). The increase in rectal temperature (P < 0.001) and respiration rate (P < 0.001) in response to the HS was greatest during lactation. There was an effect of day (P < 0.001) on serum IGF-1 and insulin concentrations because both insulin and IGF-1 increased after farrowing. Compared with HS sows, the TN sows had greater feed intake (P < 0.001) and greater serum concentrations of insulin (early lactation; P < 0.05) and IGF-1 (late lactation; P < 0.05) when they were lactating. The effects of HS on sow BW, back fat, and loin eye area were generally not significant. Average BW of individual piglets at weaning was approximately 0.5 kg lighter for the sows in the HS farrowing room (P < 0.05). Weaning-to-estrus interval, percentage sows inseminated after weaning, subsequent farrowing rate, and subsequent total born were not affected by treatment. In summary, regardless of ambient temperature, sows undergo pronounced and sustained changes in rectal temperature when they transition through gestation, lactation, weaning, and rebreeding. The effects of HS on rectal temperature, respiration rate, feed intake, and metabolic hormones were greatest during lactation. The controlled HS that we imposed affected piglet weaning weight, but rebreeding and subsequent farrowing performance were not affected.

Effect of ergot alkaloids associated with fescue toxicosis on hepatic cytochrome P450 and antioxidant proteins

Intake of ergot alkaloids found in endophyte-infected tall fescue grass is associated with decreased feed intake and reduction in body weight gain. The liver is one of the target organs of fescue toxicosis with upregulation of genes involved in xenobiotic metabolism and downregulation of genes associated with antioxidant pathways. It was hypothesized that short-term exposure of rats to ergot alkaloids would change hepatic cytochrome P450 (CYP) and antioxidant expression, as well as reduce antioxidant enzyme activity and hepatocellular proliferation rates. Hepatic gene expression of various CYPs, selected nuclear receptors associated with the CYP induction, and antioxidant enzymes were measured using real-time PCR. Hepatic expression of CYP, antioxidant and proliferating cell nuclear antigen (PCNA) proteins were measured using Western blots. The CYP3A1 protein expression was evaluated using primary rat hepatocellular cultures treated with ergovaline, one of the major ergot alkaloids produced by fescue endophyte, in order to assess the direct role of ergot alkaloids in CYP induction. The enzyme activities of selected antioxidants were assayed spectrophotometrically. While hepatic CYP and nuclear receptor expression were increased in ergot alkaloid-exposed rats, the expression and activity of antioxidant enzymes were reduced. This could potentially lead to increased oxidative stress, which might be responsible for the decrease in hepatocellular proliferation after ergot alkaloid exposure. This study demonstrated that even short-term exposure to ergot alkaloids can potentially induce hepatic oxidative stress which can contribute to the pathogenesis of fescue toxicosis.

Influence of environmental temperature on the physiological, endocrine, and immune responses in livestock exposed to a provocative immune challenge.

Although livestock experience many stressors throughout their life, one of the most commonly experienced, and most difficult to control, is stress caused by fluctuations in environmental temperatures that extend beyond the thermoneutral (TN) zone for an animal. In swine, cold stress has long been recognized as a main cause of neonatal morbidity and mortality. A possible explanation for this increased morbidity and mortality may be related to their inability to generate a febrile response. Previously, we reported that the acute phase immune response, including the generation of fever, after exposure to lipopolysaccharide (LPS; Escherichia coli O111: B4; Sigma-Aldrich, St Louis, MO, USA) is substantially altered in neonatal pigs maintained in a cold environment (ie, 18°C). Neonatal pigs that were maintained in a cold environment and administered LPS experienced a period of hypothermia coupled with altered endocrine and proinflammatory cytokine responses that could prove detrimental. In cattle, we previously reported differences in the acute phase immune response of two diverse breeds of Bos taurus cattle (Angus and Romosinuano) when maintained under TN conditions and exposed to LPS. More recently we have reported that differences in the stress and immune responses of Angus and Romosinuano heifers varies, depending on whether the cattle were housed at either TN or heat stress air temperatures. Our data clearly show that even intermittent periods of heat stress similar to that experienced in production environments can have significant effects on the stress and innate immune responses of cattle. Understanding the effect of thermal stress on livestock is critical to developing and implementing alternative management practices to improve their overall health and well-being.

Endocrine and thermoregulatory responses to acute thermal exposures in 6-month-old pigs reared in different neonatal environments

1. 1. Endocrine and thermoregulatory responses to acute heat (34°C) and cold (10°C) exposures were determined in eight pigs at 6 months of age. Half of the pigs had been reared in a cycling upper thermal environment (27–32°C) for the first 28 days of life, while the others had been reared in a lower thermal environment (21°C). 2. 2. Concentrations of cortisol increased significantly during both acute heat and cold thermal exposures (P = 0.0001) although the response was greater in the heat than in the cold (P = 0.003). A prolactin response occurred during acute heat exposure (P = 0.004). Growth hormone secretion increased during acute cold exposure (P = 0.001). There was a strong tendency for increased epinephrine secretion during both acute heat and cold exposures (P = 0.06). No significant effects of either exposure were found on plasma norepinephrine (P = 0.9), or triiodothyronine (P = 0.11). 3. 3. Neonatal environment did not affect daily core body temperature (Tc) before acute heat or cold exposures, but did alter amplitude of the 24-h Tc cycle. The amplitude was significantly greater in animals reared in the lower thermal environment (P < 0.001). 4. 4. Acute heat exposure resulted in significant increases in Tc (P < 0.001) and heat production (P < 0.01). Neonatal thermal environment had no significant effect on Tc and heat production responses to acute heat and cold thermal exposures.

Differential acute phase immune responses by Angus and Romosinuano steers following an endotoxin challenge

Our primary objective of this experiment was to evaluate potential genetic differences between two diverse Bos taurus breeds [Angus (AG) and Romosinuano (RO)] in response to an endotoxin challenge. Eighteen steers (n = 9 steers/breed; 299.4 ± 5.2 kg BW) were acclimated to environmentally controlled chambers maintained at thermoneutrality (19.7 °C) and then fitted with indwelling jugular catheters and rectal temperature (RT) recording devices 1 d before the endotoxin challenge. The next day, blood samples were collected at 30-min intervals from -2 to 8 h, and RT was measured continuously at 1-min intervals throughout the study. At time 0, all steers received an intravenous bolus injection of lipopolysaccharide (LPS; 2.5 μg/kg BW). Serum samples were stored at -80 °C until analyzed for cortisol, proinflammatory cytokines [tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), IL-6, and interferon gamma (IFN-γ)], and acute phase proteins (serum amyloid A, acid soluble protein, ceruloplasmin, and α-acid glycoprotein). Rectal temperatures increased in both breeds within 1 h after LPS, with RO producing a greater increase in RT than AG steers (P < 0.001). Serum cortisol and TNF-α increased (P < 0.01) in both breeds within 1 h after the LPS challenge. For cortisol, an overall breed effect (P < 0.02) was detected, such that AG steers had a higher cortisol response than RO steers. A breed × time interaction (P < 0.01) was observed for TNF-α, such that the response was delayed and extended in the RO steers compared with the AG steers. At 2 and 2.5 h after LPS, TNF-α concentrations were greater (P < 0.03) in RO steers than in AG steers. For IL-1β, a breed × time interaction (P < 0.04) was also observed. At 3 h after LPS, IL-1β concentrations were greater (P < 0.01) in RO steers than in AG steers. Serum IL-6 and IFN-γ increased (P < 0.01) in a similar manner in both groups after the LPS challenge. These data show differences in the innate immune response between two diverse Bos taurus breeds which may provide insight about differences observed in productivity, heat tolerance, disease resistance, and longevity among cattle breeds.

Heat-tolerant versus heat-sensitive Bos taurus cattle: influence of air temperature and breed on the metabolic response to a provocative immune challenge

The response of the immune and stress systems have been assessed in response to a lipopolysaccharide (LPS) challenge, yet the role of metabolism in mediating energy requirements during the acute phase response has not been sufficiently studied. This study tested heat-tolerant (Romosinuano [RO]) and heat-sensitive (Angus [ANG]) Bos taurus breeds at different ambient temperatures (Ta) to determine differential metabolic responses to LPS challenge. Twenty-one heifers (ANG: n = 11, 306 ± 26 kg BW; RO: n = 10, 313 ± 32 kg BW) were housed in stanchions in 4 temperature-controlled chambers. Initially, Ta in all 4 chambers was cycling at thermoneutrality (TN; 18.5°C-23.5°C) for a 1-wk adjustment period, followed by an increase in 2 chambers to cycling heat stress (HS; 24°C-38°C) for 2 wk. Five ANG and 5 RO heifers were housed at TN, whereas 6 ANG and 5 RO heifers were housed at HS. On day 19, heifers were fitted with jugular catheters. On day 20, heifers were challenged with LPS (0.5 μg/kg BW; 0 h), and blood samples were collected from -2 to 8 h and at 24 h relative to LPS challenge. Serum was analyzed for glucose, insulin, and NEFA concentrations. In addition, feed intake was measured 3 d before and on the day of the challenge. Feed intake decreased over time (P < 0.001) and was decreased in heifers housed at HS compared with heifers housed at TN (P = 0.013). Glucose concentrations before LPS challenge were greater in RO (P = 0.01) than in ANG heifers and greater in TN-housed heifers (P = 0.02) than in HS heifers. Glucose after LPS challenge initially increased before decreasing below baseline concentrations (P < 0.01) in all heifers. In addition, there was a breed by Ta interaction (P < 0.004), such that HS decreased glucose concentrations in ANG heifers compared with ANG heifers housed at TN (P < 0.001), whereas HS did not affect glucose concentrations after LPS challenge in RO heifers (P = 0.941). Nonesterified fatty acid concentrations before LPS challenge were not affected by breed (P = 0.37) or Ta (P = 0.60). Although NEFA concentration after LPS challenge was unaffected by Ta (P = 0.78), there tended to be a breed by Ta interaction (P = 0.07) such that, when housed at HS, RO heifers had greater serum NEFA concentrations after LPS challenge than ANG heifers (P = 0.009). Insulin concentration before LPS challenge was greater in RO heifers than in ANG heifers (P < 0.01). Insulin after LPS challenge increased (P < 0.01), with RO heifers producing a greater insulin response than ANG heifers (P < 0.01). These data suggest that HS decreases the metabolic response of heat-sensitive ANG heifers in response to LPS challenge, thus providing physiological evidence that may explain differences observed in the acute phase response between heat-sensitive ANG and heat-tolerant RO cattle breeds.

Effects of short-term heat stress on endophytic ergot alkaloid-induced alterations in rat hepatic gene expression

Exposure to ergot alkaloids in endophyte-infected fescue (E+) is associated with impaired animal productivity, especially during heat stress, which is commonly referred to as fescue toxicosis. To elucidate the pathogenesis of this condition, the effects of short-term heat stress (HS) on hepatic gene expression in rats exposed to endophytic ergot alkaloids were evaluated. Rats implanted with telemetric transmitters to continuously measure core temperature were fed an E+ diet and maintained under thermoneutral (TN) conditions (21 degrees C) for 5 d, followed by TN or 31 degrees C (HS) conditions for 3 d. Feed intake (FI) and BW were monitored daily. The E+ and HS-induced alterations in hepatic genes were evaluated using DNA microarrays and PCR analyses. Hepatic antioxidant enzyme activities, as well as the incidence of apoptosis, were determined. As expected, intake of E+ reduced FI and BW from pretreatment levels under TN conditions, with greater reductions during short-term HS. Genes involved in gluconeogenesis and apoptosis were upregulated, whereas genes associated with oxidative phosphorylation, xenobiotic metabolism, antioxidative mechanisms, immune function, cellular proliferation, and chaperone activity were all downregulated with short-term HS. Hepatocytic apoptosis was increased and antioxidant enzyme activity decreased in the livers of rats exposed to HS. The hypothesized, exacerbating effects of HS on the direct, endophytic toxin-related and indirect, reduced caloric intake-associated alterations in hepatic gene expression were clearly demonstrated in rats and may help to elucidate the pathogenesis of fescue toxicosis in various animal species.

Thermoregulatory response of dairy cows fed ergotized barley during summer heat stress

Claviceps purpurea infects the seed heads of cereal grains and grasses and produces ergopeptine alkaloids that cause hyperthermia and agalactia in cattle during periods of heat stress. A field experiment was undertaken to examine the effects of ergopeptine alkaloids found in barley on thermal status of dairy cattle during periods of heat stress. Production end points were also measured to identify the effect of the change in thermal status. Contaminated barley screenings containing known levels of ergopeptine alkaloids were fed to lactating Holstein cattle (10 μg total ergopeptine alkaloids/kg BW/day) for 10 days during summer heat stress. Air temperature increased 14.4 C during the first 8 days of treatment and then declined the same during the last 2 days. Extreme daily values for rectal temperature and respiration rate, using averages of all animals, showed maximum increases of 2.3 C and 56.8 breaths/minute, respectively, during this period. Group afternoon milk production decreased 2 kg/day during the heat stress period, with no measurable change in feed intake. A greater level of hyperthermia occurred in cattle consuming the diet with ergopeptine alkaloids, with only marginal symptoms of ergot toxicosis reflected in feed intake and milk production. Therefore, the ergopeptine alkaloid dose used in this study represents a level for minimal induction of the ergot toxicity response.